Giao Trinh Vi Sinh Vat Hoc

1
BIN VN MINH (CH BIN)

KIU HU NH, PHM VN TY, PHM HNG SN PHM TH NGC LAN V NGUYN TH THU THU
GIO TRNH IN T
VI SINH VT HC
Hu, nm 2008
MC LC
NHNG CH VIT TT ........................................................................................................9 CHNG 1........................................................................................................................100 VAI TR CA VI SINH VT TRONG SINH GII ..............................................................100 I. C IM CHUNG CA VI SINH VT .......................................................................100 1. Khi nim chung v vi sinh hc v vi sinh vt .........................................................100 2. Cc c im chung ca vi sinh vt. .......................................................................100 3. Cc nhm i tng vi sinh hc ...............................................................................12 4. Ni dung ...................................................................................................................13 5. Vai tr ca vi sinh vt. ...............................................................................................13 II. S LC LCH S PHT TRIN CA VI SINH VT HC ........................................14 III. CCH C (PHT M) CA CH LATINH ...............................................................24 IV. H THNG SINH GII V V TR CA CC NHM VI SINH VT ............................26 1. Khi nim v gii sinh vt .........................................................................................26 2. Mt s h thng phn loi .........................................................................................27 3. Nhng sai khc gia cc t bo Prokaryote v Eukaryote ........................................30 V. VAI TR CA VI SINH VT TRONG I SNG ........................................................31 VI. CC PHNG PHP V K THUT NGHIN CU.................................................33 Cu hi n tp chng 1 ..................................................................................................38 * Ti liu c thm ............................................................................................................38 * Ti liu tham kho ..........................................................................................................38 * Gii thch thut ng........................................................................................................38 CHNG 2..........................................................................................................................41 SINH HC CA CC C TH V BO ............................................................................41 I. KHI NIM V VIRUS, VIRION, VIROID V PRION.....................................................41 1. Virus l g?................................................................................................................41 2. c im chung ca virus.........................................................................................42 3. Virion, viroid, prion ....................................................................................................42 4. Lch s pht hin virus ..............................................................................................47 II. HNH THI V CU TRC CA VIRUS ......................................................................49 1. Hnh thi ...................................................................................................................49 2. Kch thc ca virus .................................................................................................51 3. Cu trc ca virus .....................................................................................................51 4. Genom ca virus.......................................................................................................53 III. NUI CY VIRUS .......................................................................................................54 IV. NH HNG CA VIRUS LN T BO ...................................................................55 V. CC BNH DO VIRUS ................................................................................................55 VI. NH HNG CA TC NHN VT L, HA HC N VIRUS ................................57 VII. CON NG LY NHIM VIRUS VO C TH .....................................................57 VIII. CC QU TRNH NHN LN CA VIRUS ...............................................................57
IX. CC PHNG THC NHN LN CA VIRUS .........................................................60 1. Qu trnh nhn ln ca virus RNA chui dng .......................................................60 2. Qu trnh nhn ln ca virus RNA chui n, m .....................................................62 3. S nhn ln ca virus RNA chui kp..................................................................63 4. S nhn ln ca virus Retro ................................................................................64 Cu hi n tp chng 2 ..................................................................................................65 * Ti liu c thm ............................................................................................................66 * Ti liu tham kho ..........................................................................................................66 * Gii thch thut ng.................................................................................................... 6666 CHNG 3..........................................................................................................................67 SINH HC CA CC C TH NHN S ..........................................................................67 I. C KHUN (Archaea) ...................................................................................................67 1. Khi nim .................................................................................................................67 2. c im ca c khun ............................................................................................68 3. Mi trng sng ca c khun v gi thuyt v hnh thnh s sng trn tri t .....70 4. Ph h c khun da trn trnh t 16S rRNA ...........................................................72 5. Cc hnh thc dinh dng c khun .....................................................................73 6. Mt s nhm c khun i din ................................................................................74 II. VI KHUN (BACTERIA) ...............................................................................................84 1. Khi nim chung: ......................................................................................................84 2. Lch s nghin cu v phn loi vi khun .................................................................84 4. Cu trc t bo vi khun ...........................................................................................85 5. S lc phn loi vi khun .......................................................................................92 6. Vi khun c ch v Vi khun gy hi .........................................................................94 7. Cc vn khc .......................................................................................................95 III. VI KHUN C BIT ..................................................................................................95 1. X khun (Actinomycetes) ........................................................................................95 2. Xon th (Spirochaetales) ........................................................................................98 3. Ricketsia ...................................................................................................................98 4. Mycoplasma v Chamydia ........................................................................................99 Cu hi n tp chng 3 ................................................................................................101 * Ti liu c thm ..........................................................................................................102 * Ti liu tham kho ........................................................................................................102 * Gii thch thut ng......................................................................................................103 CHNG 4........................................................................................................................104 SINH HC CA CC C TH NHN THC ...................................................................104 I. VI NM (MICROFUNGI) ..............................................................................................104 1. Nm men (Levures, Yeasts) ...................................................................................104 2. Nm mc ................................................................................................................109 3. Nm nhy ...............................................................................................................113 II. A Y ..........................................................................................................................116 III. VI TO ......................................................................................................................117
1. c im chung .................................................................................................. 11717 2. i sng ca vi to .................................................................................................118 3.Vai tr ca vi to ......................................................................................................118 IV. NG VT NGUYN SINH ......................................................................................120 Cu hi n tp chng 4 ................................................................................................122 * Ti liu c thm ..........................................................................................................122 * Ti liu tham kho ........................................................................................................122 *Gii ngha t: ................................................................................................................123 CHNG 5........................................................................................................................124 DINH DNG, SINH TRNG V SINH SN VI SINH VT .......................................124 I. THNH PHN HA HC CA T BO VI SINH VT ................................................124 1. Nc ......................................................................................................................124 2. Protein ....................................................................................................................124 3. Gluxit (hay hidrat cacbon- saccharide) ....................................................................126 4. Lipid v cc cht tng t (lipoid):..........................................................................127 5. Mt s cht hu c c hot tnh sinh hc: ..............................................................128 6. Cc nguyn t khong: ...........................................................................................129 II. DINH DNG............................................................................................................129 1. Cc cht dinh dng ..............................................................................................129 2. Cc loi mi trng sng ca vi sinh vt ................................................................129 3. Cc kiu dinh dng vi sinh vt ..........................................................................130 4. Vi sinh vt nguyn dng v khuyt dng ...........................................................131 III. SINH TRNG CA VI SINH VT ...........................................................................131 1. Cc nhn t sinh trng .........................................................................................131 2. iu kin sinh trng .............................................................................................131 3. Sinh trng trong nui cy tnh ..............................................................................131 IV. S KM HM SINH TRNG V DIT KHUN .................................................. 14040 1. Cc phng php kh trng .............................................................................. 14040 2. Kim sot tng trng ca vi sinh vt bng ha cht ......................................... 14040 3. Cc phng php bo qun ...................................................................................141 V. SINH SN VI SINH VT ........................................................................................141 1. Sinh sn vi khun (Bacteria)v c khun (Archaea) ............................................141 2. Sinh sn vi sinh vt nhn thc .............................................................................142 3. Khai thc v phng nga ca con ngi i vi VSV.............................................143 Cu hi n tp chng 5 ................................................................................................143 * Ti liu c thm ..........................................................................................................143 * Ti liu tham kho ........................................................................................................144 * Gii thch thut ng......................................................................................................144 CHNG 6........................................................................................................................146 TRAO I CHT VI SINH VT .....................................................................................146 I. NG PHN ...........................................................................................................146 II. CHU TRNH TRICARBOXYLIC ACID (Krebs) ............................................................147
III. CHUI H HP V PHOSPHORYL HA ................................................................148 1. Chui h hp ..........................................................................................................148 2. Phosphoryl ho .......................................................................................................149 IV. S VN CHUYN CHT DINH DNG VO T BO ...........................................149 V. TRAO I CHT V NNG LNG ........................................................................151 1. Cc ngun nng lng vi sinh vt .......................................................................151 2. Cc kiu h hp .................................................................................................. 15252 3. Nghin cu s trao i nng lng ........................................................................154 4. S tch tr v s dng nng lng ........................................................................156 5. S trao i carbohydrate ........................................................................................157 6. S trao i protein ..................................................................................................158 7. S trao i lipid ......................................................................................................161 Cu hi n tp chng 6 ................................................................................................162 * Ti liu c thm ..........................................................................................................162 * Ti liu tham kho ........................................................................................................162 *Gii thch thut ng.......................................................................................................163 CHNG 7........................................................................................................................164 H HP K KH ................................................................................................................164 I. KHI NIM CHUNG ....................................................................................................164 II. H HP NITRATE, AMMONIUM HA NITRITE V KH NITROGEN ......................164 III. H HP SULFATE ....................................................................................................166 IV. H HP CARBONATE TO METHANE ..................................................................167 V. H HP CARBONATE TO THNH ACETATE ........................................................169 Cu hi n tp chng 7 ................................................................................................169 * Ti liu c thm ..........................................................................................................170 * Ti liu tham kho ........................................................................................................170 * Gii thch thut ng......................................................................................................170 CHNG 8........................................................................................................................171 VI KHUN HA DNG V C HIU KH .....................................................................171 I. NITRATE HA ............................................................................................................171 1.Vi khun nitrate ha .................................................................................................171 2. C ch phn ng ca qu trnh nitrate ha. ...........................................................171 3. Cc yu t nh hng n qu trnh nitrate ...........................................................172 4. ngha ca qu trnh nitrate ha ............................................................................173 II. OXYGEN HA CC HP CHT LU HUNH .........................................................173 1. Vi khun t dng ha nng ..................................................................................173 2. Vi khun t dng quang nng ..............................................................................175 3. Vi khun d dng ..................................................................................................176 III. OXYGEN HA ST...................................................................................................176 IV. OXYGEN HA HYDROGEN.....................................................................................178 Cu hi n tp chng 8 ................................................................................................179 * Ti liu c thm ..........................................................................................................179
* Ti liu tham kho ........................................................................................................179 * Gii thch thut ng......................................................................................................179 CHNG 9........................................................................................................................181 CC QU TRNH LN MEN ..............................................................................................101 I. CC QU TRNH LN MEN PH BIN VSV ..........................................................181 II. TNH A DNG CA LN MEN .................................................................................181 III. LN MEN KHNG C S PHOSPHORYL HA C CHT .....................................182 IV. HIN TNG CNG DNG (syntrophy) ............................................................ `184 V. LN MEN RU NH NM MEN V VI KHUN .....................................................184 1. S to thnh ethanol nh nm men .......................................................................185 2. Cc dng phng trnh Neuberg ............................................................................185 3. Hiu ng Pasteur....................................................................................................186 4. K thut sn xut ethanol nh nm men................................................................187 5. Hiu sut ln men ...................................................................................................188 6. S to thnh ethanol nh vi khun .........................................................................189 VI. LN MEN LACTIC V H LACTOBACTERIACEAE........................................... 190190 1. Nhu cu v cc cht b sung v nhn t sinh trng .............................................191 2. Ln men lactic ng hnh ........................................................................................192 3. Ln men lactic d hnh .............................................................................................192 4. ng dng ca ln men lactic ..................................................................................193 Cu hi n tp chng 9 ................................................................................................193 * Ti liu c thm ..........................................................................................................197 * Ti liu tham kho ........................................................................................................197 * Gii thch thut ng : ....................................................................................................197 CHNG 10......................................................................................................................198 VI KHUN QUANG HP V C NITROGEN PHN T ..................................................198 I. VI SINH VT QUANG HP .........................................................................................198 1. Chu trnh carbon trong t nhin ..............................................................................198 2. Cc vi khun quang hp (Phototrophic bacteria) ....................................................199 3. Trao i cht cc vi sinh vt quang dng ..........................................................206 II. C NH NITROGEN.................................................................................................209 1. Vi khun c nh N cng sinh..................................................................................210 2. Vi sinh vt c nh nitrogen sng t do (khng cng sinh): .....................................212 3. Vi khun lam (xem chng III ) ...............................................................................212 Cu hi n tp chng 10 ..............................................................................................213 * Ti liu c thm ..........................................................................................................214 * Ti liu tham kho ........................................................................................................214 * Gii thch thut ng......................................................................................................214 CHNG 11......................................................................................................................216 DI TRUYN HC VI SINH VT .........................................................................................216 I. KHI NIM CHUNG ....................................................................................................216 II. C S VT CHT DI TRUYN VI SINH VT .......................................................219
1. Cu trc genom ca sinh vt nhn s ....................................................................219 2. Vt cht di truyn virus ........................................................................................222 3. Cu trc genom ca sinh vt nhn thc .................................................................224 III. C CH CHNG S XM NHP CA GENE L (hn ch v ci bin) .................225 IV. BIN D .....................................................................................................................227 1. Bin d kiu hnh .....................................................................................................227 2. Bin d kiu gene ....................................................................................................227 IV. C CH VN CHUYN VT LIU DI TRUYN .......................................................229 1. Bin np (Transformation) ......................................................................................229 2. S ti np (Transduction) .......................................................................................230 3. Tip hp (Conjugation) ...........................................................................................231 V. K THUT DI TRUYN .............................................................................................232 *Cu hi n tp chng 11 ............................................................................................235 * Ti liu c thm ..........................................................................................................234 * Ti liu tham kho ........................................................................................................235 * Gii thch thut ng......................................................................................................235 CHNG 12......................................................................................................................244 VI SINH VT GY BNH, MIN DCH HC V CC CHT C HOT TNH SINH HC244 I. VI SINH VT GY BNH ............................................................................................244 1. Bnh truyn nhim v khng truyn nhim .............................................................244 2. c lc v c t ....................................................................................................245 3. Bnh do virus, vi khun v cc phn t hu c khc. ............................................245 II. KHNG NGUYN, KHNG TH V B TH ............................................................247 1. Khng nguyn ........................................................................................................247 2. Khng th ...............................................................................................................248 3. B th .....................................................................................................................254 III. VACCINE V HUYT THANH MIN DCH ...............................................................256 1. Vaccine...................................................................................................................256 2. Huyt thanh ............................................................................................................257 IV. MIN DCH ...............................................................................................................258 1. nh ngha: .............................................................................................................258 2. C ch bo v khng c hiu ...............................................................................258 3. Cht sinh min dch (immunogen) ..........................................................................260 4. Tnh c hiu ca khng nguyn (KN) ....................................................................260 5. Cc c quan v t bo tham gia vo h thng min dch ........................................260 6. Min dch bnh l ....................................................................................................262 V. CC CHT C HOT TNH SINH HC ....................................................................266 1. Enzym ....................................................................................................................266 2.Cht khng sinh .......................................................................................................268 3. Vitamin ...................................................................................................................275 Cu hi n tp chng 12 ..............................................................................................276 * Ti liu c thm ..........................................................................................................276
* Ti liu tham kho ........................................................................................................276 * Gii thch thut ng......................................................................................................276 CHNG 13......................................................................................................................279 SINH THI HC VI SINH VT ...........................................................................................279 I. I CNG V SINH THI HC VI SINH VT .........................................................279 1. Cc khi nim c bn .............................................................................................279 2. c im ca vi sinh vt trong t nhin ..................................................................279 3. Cc phng php nghin cu sinh thi hc vi sinh vt...........................................280 4. Hot ng v vai tr ca vi sinh vt trong cc h sinh thi ......................................285 5. Vai tr ca VSV trong cc chu trnh sinh i ha - cc nguyn t cn cho s sng 288 II. S PHN B CA VI SINH VT TRONG T NHIN ...............................................298 1. Vi sinh vt trong khng kh......................................................................................398 2. Vi sinh vt t .........................................................................................................299 3. Vi sinh vt nc......................................................................................................299 III. MT S BIN PHP VI SINH GP PHN BO V MI TRNG ........................300 1. Nguyn nhn gy nhim mi trng ....................................................................300 2. Mt s bin php vi sinh gp phn bo v mi trng............................................301 3. Mt s bin php hin ang c p dng trong nc v trn th gii ..................301 *Cu hi n tp chng13 .............................................................................................311 * Ti liu c thm ..........................................................................................................312 * Ti liu tham kho ........................................................................................................312 * Gii thch thut ng......................................................................................................312 CHNG 14.................................................................................................................. 31313 CU HI N TP .......................................................................................................... 31313 I. PHN CU HI....................................................................................................... 31313 II. TR LI .....................................................................................................................319 TI LIU THAM KHO ......................................................................................................321
Gio trnh in t ny c bin son li da trn gio trnh Vi sinh vt hc, NXB i hc Hu (2006) ca nhm tc gi: Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu.
NHNG CH VIT TT
ADP AMP APG
A-1,3DPG
Adenosine diphosphate Adenosine monophosphate Acid 3-phosphoglyceric Acid 1,3 diphosphoglyceric Adenosine triphosphate Acid 6-penicillanic Coenzym A Cht khng sinh DeOxygenribonucleic acid Ribulose-1,5-diphosphate Ribulose-5-diphosphate Ribonucleic acid Vi sinh vt Fructose-6-phosphate Flavin adenine dinucleotide Glucose-6-phosphate Glyceraldehyde phosphate 2-Keto-3-deOxygen-6-phosphogluconate Nitrogen Nicotinamid adenine dinucleotide dng Oxygen ha Nicotinamid adenine dinucleotide dng kh
Nicotinamid adenine dinucleotide phosphat dng Oxygen ha
ATP A-6PA CoA CKS DNA R-1,5-DP R-5-P RNA VSV F-6-P FAD G-6-P GAP KDPG N NAD NADH NADP NADPH PP X-5-P
Nicotinamid adenine dinucleotide phosphat dng kh Pentose phosphate Xylulose-5-phosphate
Chng 1: Vai tr ca vi sinh vt trong sinh gii
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CHNG 1
VAI TR CA VI SINH VT TRONG SINH GII

I. C IM CHUNG CA VI SINH VT
1. Khi nim chung v vi sinh vt hc v vi sinh vt

1.1. Vi sinh vt hc (Microbiology) L khoa hc nghin cu cc c th sng hin vi v siu hin vi, cu to t bo v quy lut hot ng ca chng, s dng cc vi sinh vt nhm phc v li ch ca con ngi v gi vng h sinh thi trn Tri t. 1.2. Vi sinh vt (Microorganisms) L tn gi chung ch tt c cc loi sinh vt nh b, ch c th nhn r di knh hin vi quang hc hoc knh hin vi in t. Vi sinh vt bao gm nhiu nhm khc nhau: 1. Virus 2. Archaea (C khun hay cn gi l vi khun c) 3. Vi khun (Bacteria) 4. X khun (Actinomycetes) 5. Vi nm (Microfungi) 6. Vi to (Microalgae) Gia cc nhm trn khng c mi lin h cht ch v mt hnh thi hay phn loi, nhng ngi ta gp chng li v chng cng c mt s phng php nui dng, nghin cu v hot ng sinh l gn ging nhau.
2. Cc c im chung ca vi sinh vt.

2.1. Kch thc nh b Vi sinh vt thng c o kch thc bng n v micromet (1m= 1/1000mm hay 1/1 000 000m).Virus c o kch thc n v bng nanomet (1nm=1/1 000 000mm hay 1/1 000 000 000m).
Hnh 1.1: Cc phng php quan st th gii sng (t nguyn t n t bo)
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2.2. Hp th cht dinh dng trc tip qua b mt t bo, chuyn ha nhanh a s VSV l n bo nn chng nhn cc cht dinh dng bng hp th (absorbtion) qua b mt t bo, khc vi thc vt l t dng (autotrophic) v ng vt l ni tiu ha (ingestion) qua ng tiu ha. Chnh iu ny m vic nui cc VSV c thc hin d dng v nhanh chng. Mt vi khun lctic (Lactobacillus) trong 1 gi c th phn gii c mt lng ng lactose ln hn 100-10 000 ln so vi khi lng ca chng. Tc tng hp protein ca nm men cao gp 1000 ln so vi u tng v gp 100 000 ln so vi tru b. 2.3. Kh nng sinh sn nhanh Thi gian th h ngn. - 1 trc khun Escherichia coli trong cc iu kin thch hp ch sau 12-20 pht li phn ct mt ln. - Nm men ru (Saccharomyces cerevisiae) l 120 pht. - To Tiu cu ( Chlorella ) l 7 gi, vi vi khun lam Nostoc l 23 gi...
Vi khun Escherichia coli
Nm men S. cerevisiae
Nm si Alternaria sp.
Vi to Chlorella sp.
Hnh 1.2. Mt s vi sinh vt c s dng trong i sng 2.4. Kh nng thch ng rt cao v pht sinh bin d mnh
Vi sinh vt a s l n bo, n bi, sinh sn nhanh, s lng nhiu, tip xc trc tip vi -5 -10 mi trng sng. Do , rt d dng pht sinh bin d. Tn s bin d thng mc 10 -10 .
V d. Khi mi pht hin ra penicillin hot tnh ch t 20 n v/ml dch ln men (1943) th nay c th t trn 100 000 n v/ml. 2.5. Phn b rng, chng loi nhiu Vi sinh vt c mt khp mi ni trn Tri t, trong khng kh, trong t, trn ni cao, di bin su, trn c th, ngi, ng vt, thc vt, trong thc phm, trn mi vt... Ngi ta c tnh trong s 1,5 triu loi sinh vt c khong 200 000 loi vi sinh vt (100 000 loi ng vt nguyn sinh v to, 90 000 loi nm, 2500 loi vi khun lam v 1500 loi vi khun). Tuy nhin hng nm, c thm hng nghn loi sinh vt mi c pht hin, trong c khng t loi vi sinh vt. 2.6. S a dng ca cc phn ng ha sinh hc Cc phn ng sinh ha trong c th VSV thng n gin hn nhiu so vi trong c th ng, thc vt. Nhng mi loi c mt s phn ng ring nn cc phn ng sinh ha ca
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cc loi VSV khc nhau rt a dng. D mt hp cht c phc tp n u, trong thin nhin u c cc VSV s dng hoc phn hy chng. Sn phm do loi ny to ra c th c loi khc s dng. Mi loi thng to ra mt s cht trao i th cp (secondary metabolites) c hiu gip cho chng pht trin tt hn v km hm mt s loi khc. V d: cc loi nm men ru thch nghi vi nng ng cao v to ra ru l cht hn ch s pht trin nhiu loi khc. Do c im ny, sn phm khi b nhim s km hm s tng trng ca cc chng sn xut. 2.7. C chng xut hin sm nht trn tri t Tri t hnh thnh cch y khong 4,6 t nm nhng ch tm thy du vt ca s sng t cch y 3,5 t nm. Vi sinh vt ho thch c xa nht c pht hin l nhng dng rt ging vi vi khun lam ngy nay. Chng c J.William Schopf tm thy ti cc tng c min Ty Australia. Chng c dng a bo n gin, ni thnh si di n vi chc mm vi ng knh khong 1-2 mm v c thnh t bo kh dy. Trc cc nh khoa hc cng tm thy vt tch ca chi Gloeodiniopsis c nin i cch y 1,5 t nm v vt tch ca chi Palaeolyngbya c nin i cch y 950 triu nm.
Vt tch vi khun lam Cyanobacteria cch y 3,5 t nm
Vt tch Gloeodiniopsis cch y 1,5 t nm
Vt tch Palaeolyngbya cch y 950 triu nm
Hnh 1.3. Cc vi sinh vt ho thch
3. Cc nhm i tng vi sinh vt hc

Vi sinh vt hc hin i i su nghin cu tng nhm i tng ring bit trn v tr thnh nhng mn hc chuyn su nh: virus hc (Virology) - nghin cu vi sinh vt cha c cu to t bo (virus RNA v virus DNA), vi khun hc (Bacteriology) - nghin cu VSV nhn s, gm c khun (Archaea) v vi khun (Bacteria), nm hc (Mycology)- nghin cu cc vi nm (nm men, nm si hay nm mc v nm nhy), to hc (Algology)- nghin cu cc vi to v ng vt nguyn sinh hc(Protozoology)- nghin cu cc ng vt nguyn sinh. Mt khc vi sinh vt hc hin i cng i su nghin cu nhng tnh cht ring bit ca VSV v hnh thnh cc chuyn ngnh nh t bo hc, phn loi hc, sinh l hc, ha sinh hc, di truyn hc ca vi sinh vt.
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V mt ng dng ngnh vi sinh vt hc gm c cc chuyn ngnh nh: vi sinh vt hc cng nghip, vi sinh vt hc thc phm, vi sinh vt hc y hc, vi sinh vt hc th y, vi sinh vt t, vi sinh vt hc nc, vi sinh vt hc khng kh, vi sinh vt hc du ha...v ngy nay cn thm ngnh vi sinh vt hc ngoi tri t (Exomicrobiology).
4. Ni dung
Lch s ra i v pht trin ca ngnh vi sinh vt hc. Vai tr ca vi sinh vt trong sn xut v i sng. Cu to v phn loi vi sinh vt. Sinh l vi sinh vt. Di truyn vi sinh vt (virus, c khun, vi khun v vi nm). Vi sinh vt gy bnh, min dch hc v cc cht c hot tnh sinh hc. nh hng ca iu kin ngoi cnh n hot ng ca vi sinh vt. S phn b vi sinh vt trong t nhin.
5. Vai tr ca vi sinh vt.
Hnh 1.4: Mt s ch li ca VSV trong nng nghip, thc phm i a s vi sinh vt l bn: - V nng nghip: c nh m cho cy trng; tun hon cc cht dinh dng trong t; gip gia sc tiu ha c, rm thnh tht - V thc phm: to cc thc phm ln men (bia, ru, fomage, yaourt); ko di thi gian bo qun; to cc ph gia thc phm - V cng nghip: to ra cc dung mi hu c, cc cht dinh dng, vitamin, sinh khi - V y t: sn xut khng sinh, gip n nh h vi khun ng rut - V mi trng: phn hy cc cht thi, ci thin mi trng b nhim thuc tr su, thuc dit c
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- V nng lng: to kh methane dng lm nhin liu; to H2 t nng lng nh sng v cc ngun nng lng v c, hu c dng lm ngun nng lng ti sinh ca tng lai. - C vai tr khng th thiu trong Cng ngh Sinh hc hin i. Mt s t vi sinh vt l th: - Gy bnh trn ngi - Gy bnh trn vt nui - Gy bnh trn cy trng. - Gy h hng cc dng c thit b
Hnh 1.5. ng dng ca vi sinh vt trong cng nghip
II. S LC LCH S PHT TRIN CA VI SINH VT HC

Nm 1546, Girolamo Fracastoro (1478- 1553) cho rng cc c th nh b l tc nhn gy ra bnh tt.
Nm 1590, Zacharias Janssen (1580-1638) ngi H Lan u tin lp ghp knh hin vi.
Nm 1665, Robert Hooke nh khoa hc ngi Anh, ln u tin quan st thy t bo sng di knh hin vi ming bc bn v a ra khi nim: T bo Cell
Hnh 1.6: 1.Girolamo Fracastoro (1478-1553); 2.Zacharias Janssen (1580-1638); 3. Robert Hooke (1635 - 1703) v 4. Lt ct ming bc bn
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Nm 1676, Antony van Leeuwenhoek(162-1723) hon thin knh hin vi v khm ph ra th gii vi sinh vt (m ng gi l anmalcules).
[A]
[B]
[C]
Hnh 1.7: Antony van Leeuwenhoek (162-1723) [A] Knh hin vi u tin ca nhn loi [B] Bt tch miu t vi sinh vt ca Leeuwenhoek [C] Nm 1688, nh vn vt hc ngi Francisco Redi (1626-1697) cng b nghin cu v s pht sinh t nhin ca gii. Nhng nm 1765-1776, Spallanzani (1729-1799) cng kch thuyt Pht sinh t nhin. Nm 1798, Edward Jenner (1749-1823) ngh ra phng php chng m u b phng nga bnh u ma. Nhng nm 1838-1839, Theodor Schwann (1810-1882) v Matthriat Schleiden (1804-1881) cng b Hc thuyt t bo.
Hnh 1.8: 1.Francisco Redi (1626-1697); 2.Spallanzani (1729-1799); 3.Theodor Schwann (1810-1882) v 4. Matthriat Schleiden (1804-1881) Nhng nm 1847-1850, Ignaz Philipp Semmelweis (1818-1865) cho rng bnh st hu sn ly truyn qua thy thuc v kin ngh dng phng php v khun phng bnh. Nm 1873, Gerhard Armauer Hansen (1841-1912) tm thy trc khun Mycobacterium leprae gy bnh hi.
Nm 1875 Ferdinand Julius Cohn (1828-1898) tm c vi khun than Bacillus anthracis.
Nm 1880, Charles Louis Alphonse Laveran (1845 -1922) pht hin k sinh trng Plasmodium gy ra bnh st rt.
16
1 2 3 4 Hnh 1.9: 1.Semmelweis (1818-1865); 2. G. A. Hansen (1841-1912) ; 3. F. J. Cohn (1828-1898) ; 4.Charles Louis Alphonse Laveran (1845 -1922) Ngi c cng ln nht khai sinh ra vi sinh vt hc thc nghim, nhm nghin cu cc hot ng sinh l, sinh ho ca vi sinh vt v ng dng chng trong ln men, c bit trong ch to vaccine phng bnh di, l nh bc hc li lc ngi Php Louis Pasteur (18221895). ng thi v tip theo Pasteur cng c nhiu nh vi sinh hc ni ting: Robert Koch (1843-1910) nghin cu vi khun gy bnh lao (Mycobacterium tuberculosis-1882), bnh t ( Vibrio cholerae-1883), ng sng to nhiu phng php nghin cu nh k thut c nh, nhum mu vi khun, nui cy v phn lp VSV trn mi trng c. Nm 1884, Elie Metchnikoff (1845-1916) miu t hin tng thc bo (phagocytosis); Hans Christian J. Gram (1853-1938) tm ra phng php nhum Gram.
1 2 3 4 Hnh 1.10: 1. Louis Pasteur (1822-1895); 2. Robert Koch (1843-1910); 3. Elie Metchnikoff (1845-1916); 4. Theodor Escherich (1857 1911) Nm 1885, Theodor Escherich (1857 1911) tm ra vi khun Escherichia coli gy ra bnh tiu chy; Daniel E. Salmon (1850-1914) pht hin ra Salmonella typhi gy ra bnh thng hn. Nm 1886, Fraenkel pht hin thy Streptococcus pneumoniae gy ra bnh vim phi.
17
Nm 1887, Richard Petri (1852-1921) pht hin ta cch dng hp lng (a Petri) nui cy vi sinh vt. Nhng nm 1887-1890, Serge Winogradsky (1856-1953) nghin cu v vi khun lu hunh v vi khun nitrate ho.
1 2 3 4 Hnh 1.11: 1. Daniel E. Salmon (1850-1914); 2. Hans Christian J. Gram (1853-1938); 3. Richard Petri (1852-1921); 4. Serge Winogradsky (1856-1953) Nm 1889, Martinus Beijerinick (1851-1931) phn lp c vi khun nt sn t r u. Nm 1890, Emil Adolph von Behring (1854-1917) lm ra khng c t chng bnh un vn v bnh bch hu. Nm 1892, Dmitri Iwanowski (1864-1920) pht hin ra mm bnh nh hn vi khun (virus) gy ra bnh khm cy thuc l. Nm 1894, Alexandre Yersin (1863-1943) v Kitasato Shibasaburo (1852-1931) khm ph ra vi khun gy bnh dch hch (Yersina pestis).
1 2 3 4 Hnh 1.12: 1. Martinus Beijerinick (1851-1931); 2. Emil Adolph von Behring (1854-1917) 3. Dmitri Iwanowski (1864-1920) ; 4. Alexandre Yersin (1863-1943) Nm 1895, Jules Bordet (1870-1961) khm ph ra B th (complement) Nm 1896, Emile van Ermengem tm ra mm bnh ng c tht (vi khun Clostridium botulinum). Nm 1897, Eduard Buchner (1860- 1917) tch ra c cc men (ferments) t nm men (yeast); Ross, Sir Ronald (1857-1932) chng minh k sinh trng st rt ly truyn bnh qua mui.
18
Nm 1899, Martinus Beijerinick (1851-1931) chng minh nhng ht virus gy nn bnh khm l thuc l.
1 2 3 4 Hnh 1.13: 1. Jules Bordet (1870-1961); 2. Emile van Ermengem(?); 3. Ronald Ross(1857-1932); 4. Eduard Buchner (1860- 1917)
Nm 1900, Major Walter Reed (1851-1902) chng minh bnh st vng ly truyn do mui.
Nm 1902, Karl Landsteiner (1868-1943) khm ph ra cc nhm mu. Nm 1903, Wright v cng s khm ph ra khng th (antibody) trong mu ca cc ng vt min dch. Nm 1905, Fritz Schaudinn (1871-1906) v Jakob Wassermann (1873-1934) tm ra mm bnh giang mai (Treponema pallidum).
1 2 3 4 Hnh 1.14: 1. Major Walter Reed (1851-1902); 2. Karl Landsteiner (1868-1943); 3. Fritz Schaudinn (1871-1906); 4. Jakob Wassermann (1873-1934) Nm 1906, Jakob Wassermann pht hin ra xt nghim c nh b th chn on giang mai. Nm 1909, Howard Taylor Ricketts (1871-1910) chng minh bnh St ban ni lan truyn qua ve l do mm bnh vi khun (Rickettsia rickettsii). Nm 1910, Peyton Rous (1879-1970) pht hin ra ung th gia cm.
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1 2 3 4 Hnh 1.15: 1. Jakob Wassermann (1873-1934); 2. Ricketts (1871-1910); 3. Prowazek(1875-1915) v 4. Peyton Rous (1879-1970) Nm 1915, Frederick Twort (1877-1950) v nm 1917, Felix d'Herelle (1873-1949) pht hin ra virus ca vi khun ( thc khun th-phage). Nm 1923, xut bn ln u cun Phn loi vi khun (Bergey's Manual of Systematic Bacteriology) Nm 1928, Frederick Griffith ( 1881-1941) khm ph ra vic bin np (transformation) vi khun.
1 2 3 4 Hnh 1.16; 1. Frederick Twort (1877-1950); 2. Felix d'Herelle (1873-1949); 3. David Hendricks Bergey (1860-1937); 4. Frederick Griffith ( 1881-1941) Nm 1928, Alexander Fleming (1881-1955) pht hin ra cht khng sinh penicillin. Nm 1931, Van Niel (1897-1985) chng minh vi khun quang hp s dng cht kh nh ngun cung cp electron v khng sn sinh oxygen. Nm 1933, Ernst August Friedrich Ruska (1906-1988) lm ra chic knh hin vi in t u tin. Nm 1935, Wendell Stanley (1904-1971) kt tinh c virus khm thuc l (TMV); Gerhard Domag (1895 1964) tm ra thuc sulfamide. Nm 1937, Edouard Chatton (1883-1947) phn chia sinh vt thnh hai nhm: Nhn s (Procaryotes) v Nhn thc (Eukaryotes).
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1 2 3 4 Hnh 1.17: 1. Alexander Fleming (1881-1955); 2. Friedrich Ruska (1906-1988); 3. Wendell Stanley (1909-1971) v 4. Gerhard Domag (1895 1964) Nm 1941, George W. Beadle (1903-1989) v Tatum, Edward Lawrie (1909-1975) a ra gi thuyt mt gen- mt enzym. Nm 1944, Oswald Avery (1877-1955) chng minh DNA chuyn thng tin di truyn trong qu trnh bin np; Selman Abraham Waksman(1888-1973) tm ra cht khng sinh streptomycin.
1 2 3 4 Hnh 1.18: 1. Edouard Chatton (1883-1947); 2.George W. Beadle (1903-1989); 3. Tatum, Edward Lawrie (1909-1975); 4. Oswald Avery (1877-1955). Nm 1946, Lederberg Joshua (1925-?), v Tatum Edward Lawrie (1909-1975), khm ph ra qu trnh tip hp (conjugation) vi khun. Nm 1949, Enders, John Franklin (1897-1985), Weller, Thomas Huckle (1915- ) v Robbins, Frederick Chapman (1916-2003), nui c virus Polio (Poliovirus) trn m ngi nui cy v nhiu loi m khc nhau. Nm 1950, Andr Michel Lwoff (1902-1994) xc nh c cc thc khun th tim tan (lysogenic bacteriophages). Nm 1951 Barbara McClintock (1902-1992) pht hin ra gen nhy.
21
Hnh 1.19: 1.Selman Abraham Waksman (1888 1973); 2. Barbara McClintock (1902-1992); 3. Lederberg Joshua (1925- ); 4. F.C.Robbins (1916-2003) Nm 1952, Alfred Hershey v Martha Chase chng minh thc khun th tim DNA ca mnh vo t bo vt ch (host); Zinder v Lederberg khm ph ra qu trnh ti np (transduction) vi khun. Nm 1953, Watson v Crick khm ph ra chui xon kp ca DNA; Frits (Frederik) Zernike (1888-1966) lm ra knh hin vi tng phn pha (phase-contrast microscope); Medawar khm ph ra hin tng nhn min dch (immune tolerance).
Hnh 1.20: 1. A.M.Lwoff(1902-1994); 2. Alfred Hershey v Martha Chase; 3. J.D.Watson (1928-) v 4. F.H.C.Crick (1916-2004) Nm 1955, Francois Jacob (1920- ) v Jacques Monod (1910-1976) khm ph ra yu t F l mt plasmid; Jerne v Burnet chng minh l thuyt chn lc clone (clonal selection). Nm 1959, Yalow trin khai k thut min dch phng x. Nm 1961, Jacob v Monod gii thiu m hnh iu ho hot ng gen nh operon. Nhng nm 1961-1966, Khorana, Har Gobind (1922-) Nirenberg Marshall (1927-) v cng s gii thch m di truyn.
22
1 2 3 4 Hnh 1.21 : 1. Francois Jacob (1920-); 2. Jacques Monod (1910-1976); 3. Har Gobind Khorana(1922-); 4.Nirenberg Marshall (1927- ) Nm 1962, Porter chng minh cu trc c bn ca Globulin min dch G. Nm 1970, Nathans Daniel (1928-1999), Arber Werner (1929-), Smith Hamilton O. (1931-) khm ph ra enzym gii hn (restriction endonuclease); Temin v Baltimore khm ph ra enzym phin m ngc (reverse transcriptase)
1 2 3 Hnh 1.22. 1.Nathans, Daniel (1928-1999); 2. Arber Werner (1929-); 3. Smith Hamilton O. ( 1931-) Nm 1973, Ames trin khai phng php vi sinh vt hc khm ph ra cc yu t gy t bin (mutagens); Cohen, Boyer, Chang v Helling s dng vector plasmid tch dng gen vi khun.
1 2 3 4 Hnh 1.23: 1. Temin, Howard Martin(1934-1994); 2.Paul Berg (1926-); 3.Stanley N. Cohen (1935-); 4. Herbert Boyer (1936-)
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Nm 1975, Kohler v Milstein pht trin k thut sn xut cc khng th n dng ( monoclonal antibodies). Nm 1977, Woese v Fox tha nhn Vi khun c (Archaea) l mt nhm vi sinh vt ring bit; Walter Gilbert v Frederick Sanger trin khai k thut gii trnh t DNA (DNA sequencing) Nm 1979, tng hp Insulin bng k thut ti t hp DNA. Nm 1980, pht trin knh hin vi in t qut Nm 1981, Stanley B. Prusiner(1942- ) pht hin ra cc c trng ca Prion Nm 1982, pht trin vaccin ti t hp chng vim gan B. Nhng nm 1983-1984, Luc Montagnier phn lp v nh loi virus gy suy gim min dch ngi Nm 1986, ln u tin ng dng trn ngi vaccin c sn xut bng k thut di truyn (vaccin vim gan B). Nm 1990, bt u th nghim ln u tin liu php gen (gene-therapy) trn ngi. Nm 1995, Venter, Smith gii trnh t h gen ca vi khun Haemophilus influenzae. Nm 1996, gii trnh t h gen ca vi khun Methanococcus jannaschii; Gii trnh t h gen nm men S. cerevisiae gm 6 000 gene. Nm 1997, pht hin ra loi vi khun ln nht Thiomargarita namibiensis; Gii trnh t h gen vi khun Escherichia coli c 3 200 gene. Nm 2000, Edward Delong pht hin ra Archea bin. Vi sinh vt hc l mt ngnh khoa hc c tc pht trin mnh m, nhiu gii thng Nobel c trao cho cc nh vi sinh vt hc hoc nhng cng trnh nghin cu trn i tng vi sinh vt. Gii Nobel Y hc 2005 dnh cho hai nh khoa hc Australia - Barry J. Marshall v J. Robin Warren - do khm ph ra vi khun Helicobacter pylori v vai tr ca chng trong bnh vim lot h tiu ho (chng vim d dy, lot d dy hoc t trng).
2 3 Hnh 1.24: 1.Barry J. Marshall (1951-); 2. J. Robin Warren (1937-) v vi khun Helicobacter pyroli sng trong y d dy
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Gii Nobel Y hc 2006 dnh cho hai nh khoa hc M Andrew Z. Fire v Craig C. Mello ca M c cng khm ph cch v hiu ha nh hng ca cc gene c bit.
1 2 3 Hnh 1.25: 1. Craig C. Mello (1960-); 2. Andrew Z. Fire (1959-) v c ch v hiu ha gen Gii Nobel Y hc nm 2008 c trao cho nh khoa hc c Harald zur Hausen tm ra cc virus papilloma (HPV) gy bnh ung th c t cung ph n v hai nh khoa hc Php Barre-Sinoussi v Luc Montagnier khm ph virus suy gim min dch ngi (HIV), gy bnh AIDS.
1 2 3 4 Hnh 1.26: 1. Harald zur Hausen ,2. Francoise Barre-Sinoussi, 3. Luc Montagnie, 4.Virus papilloma (HPV) thuc nhm dsDNA Ngy nay vi sinh vt hc pht trin rt su vi hng trm nh bc hc c tn tui v hng chc ngn ngi tham gia nghin cu. Cc nghin cu i su vo bn cht ca s sng mc phn t v di phn t, i su vo k thut cy m v tho lp gene vi sinh vt v ng dng k thut tho lp ny cha bnh cho ngi, gia sc, cy trng v ang i su vo gii quyt dn bnh ung th loi ngi.
III. CCH C (PHT M) CA CH LATINH

Ch Latinh tuy khng cn thng dng trn th gii nhng cc nh khoa hc vn s dng loi ch ny t tn cho cc loi vi sinh vt. Chng ta cn bit cch c loi ch ny pht m cho ng, nh th ngi khc mi hiu c khi ta trao i vi h bng bt c loi ngn ng no.
25
Bng 1.1: Cch c (pht m) ca ch Latinh

STT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 Ch ci A B C D E F G H I J K L M N O P Q R S T U V X Y Z W a b c d e f g h i j k l m n o p q r s t u v x y z w Tn gi a b x p-ph gh ht i ita ca e l em- m en-n p cu e-r t-x t u v ich-x d-ta a b k(cng) x (mm) ph gh h i i k l m n p q r x t u v kx d v u beta cicada decem ego familia gutta homo iodum kalium lamina mel nomen ovum penna raus semen tela Cch c apis Th d a-pi-x (ong) b-ta (c ci ng) xi-ca-da (ve su) - xm (mi) -g (ti) pha-mi-lia (h) gu-ta (git) h-m (ngi) i--um (it) ca-li-um (kali) la-mi-na (phin l) m- l (mt ong) n-mn (tn) -vum (trng) pn-na (lng chim) ra-ru-x (him) x-mn (ht) t- la (vi)
injectio in-ch-xi- (thuc tim)
quinque quanh-qu (nm)
uo, dvo u-(hai), c th vit:dvo vox simples zona v-ch- x (ting ni) xim-plch- x (n gin) d- na (vng)
ip-xi-lon uy
amylum a-my-lum (tinh bt) Wolfram vn-ph-ram wedelia u---li-a (cy si t)
Bng 1.2. Quy tc vn phm Latinh bin i t s t sang s nhiu

Ging S t S nhiu Th d Ci -a -ae alga, algae c -us -i fungus, Fungi Trung -um -a Bacterium, Bacteria
26
IV. H THNG SINH GII V V TR CA CC NHM VI SINH VT

1. Khi nim v gii sinh vt
Gii (Kingdom) l n v phn loi hin nay bao gm nhng sinh vt c chung nhng c im nht nh. Cc h thng phn loi sinh vt l kt qu ca hn 200 nm nghin cu v h thng hc. H thng cc c th ngy cng hp l nh nhng hiu bit su sc v sinh hc phn t. Ngy nay nh cc phng php phn loi hin i nh: Ha phn loi (Chemotaxonomy), phn loi s (Numerical taxonomy), phn loi chng loi pht sinh (Phylogeney taxonomy),... m khoa hc xc nh v tr kh chnh xc ca cc nhm c th v mi lin h chng loi pht sinh gia chng. Th gii sinh vt v cng phong ph v a dng. nghin cu chng cc nh khoa hc phi da vo cc tiu ch v cu tao, dinh dng, sinh sn... sp xp chng vo bc thang phn loi v t tn Cc sinh vt c sp xp theo thang phn loi l thuc t thp n cao: Loi (Species), Chi (Genus), H (Family), B (Order), Lp (Class), Ngnh (Phylum), v Gii (Kingdom). Hin nay trn gii cn c mt mc phn loi na gi l lnh gii (Domain). y l cha k n cc mc phn loi trung gian nh Loi ph (Subspecies), Chi ph (Subgenus), H ph (Subfamily), B ph (Suborder), Lp ph (Subclass), Ngnh ph (Subphylum). Loi l bc thang phn loi thp nht, lnh gii l cp phn loi cao nht. Bt k mt sinh vt no cng c sp xp vo mt loi nht nh. Nhiu loi thn thuc tp hp thnh chi, nhiu chi thn thuc tp hp thnh h, nhiu h thn thuc tp hp thnh b, nhiu b thn thuc tp hp thnh lp, nhiu lp tp hp thnh ngnh, nhiu ngnh hp thnh mt gii. trnh nhm ln ngi ta t tn loi theo nguyn tc dng tn kp (theo ting Latinh). Tn th nht l tn chi (vit hoa), tn th 2 l tn loi (vit thng) V d: Escherichia coli. Khi cn vit tt ta ch vit tt tn chi, tn loi vit y (bng ch thng). V d: E. coli . Mt loi bt k th thng vit tt l "sp." cn s nhiu l "spp.".
2 3 Hnh 1.27 : 1. A. P. de Candolle (1778 -1841), 2. George Gaylord Simpson ( 1902 1984) v 3. Ernst Mayr(1904-2005)
27
2. Mt s h thng phn loi

Bng 1.3. Mt s h thng phn loi sinh vt
*Trc Linne c nhiu tc gi phn loi sinh vt ni ting, ng ch l Aristotle (384-322 TCN) c nhiu ng gp trong phn loi ng vt.
Hnh 1.28. Phn loi th gii sng
28
Hnh 1.29: Mt s nh phn loi hc xut sc (t tri qua phi) Carolus Linnaeus (1707-1778); ; Ernst Heinrich Haeckel (1834-1919); Robert Whittaker (1921-1981) v Carl R. Woese (1928-)
Hnh 1.30: H thng 3 lnh gii ca Carl R. Woese v cng s, 1990
29
Hnh 1.31: V tr phn loi ca c khun v h thng ba lnh gii (Domain) Carl R. Woese, 1990
Hnh 1.32. H thng phn loi 8 gii sinh vt (T. Cavalier- Smith -1993) Bng 1.4: H thng ba siu gii vi by gii* (dn theo Nguyn Thnh t, 2001)
TT 1 2 3
*
Siu gii Acaryote Prokaryote Eukaryote
Cc gii sinh vt Protovira (Virus RNA); Euvira(Vi rut DNA) Archaea (Vi sinh vt c); Bacteria (Vi khun) Plantae (Thc vt); Fungi (Nm); Animalia (ng vt)
Cc siu gii ny c sp xp theo 3 mc phc tp ca c th sng
30
3. Nhng sai khc gia cc t bo Prokaryote v Eukaryote

Nhm sinh vt nhn s v nhn thc c nhng sai khc c bn (Bng 1.5): Bng 1.5: Nhng sai khc gia cc t bo Prokaryote v Eukaryote
c im Sinh vt in hnh *T chc di truyn: Cu trc nhn t bo -S NST -Cc NST cha histon -Hch nhn -Trao i di truyn *Cc cu trc ca t bo: -Li ni cht -B my golgi -Ti th -Cc lisosom -Cc lp th -Kch thc ribosom -Si thoi v sc -Mng sinh cht khng c khng c khng c khng c khng c 70s khng c khng cha steron(tr vk lam) c c c c c thc vt 80s nhn,70s -bo quan c c cha steron vng nhn; khng c cu trc in hnh 1 khng c khng c 1 chiu qua plasmid cu trc nhn in hnh vi mng nhn c cc cu trc l nhn >1 c c bng s kt hp giao t Nhm nhn s (Prokaryote) bacteria, archaea Nhm nhn thc (Eukaryote) protista, nm, ng vt, thc vt
-Vch t bo cha PG c murein ngoi tr mycoplasma khng c (Peptidoglycan) v vk c *Chc nng c trng: -Thc bo -m bo -Dng t bo cht khng c khng c khng c chuyn ng ni bo i khi c i khi c mng bo quan chuyn ng ni bo r rt
-V tr vn chuyn in t mng t bo
Ngoi virus l nhm vi sinh vt cha c cu trc t bo ra, tt c cc sinh vt khc trn Tri t ny c chia thnh hai nhm ln: Nhm nhn s (Prokaryote): bao gm vi khun (theo ngha rng gm vi khun (Bacteria), x khun (Actinomycetes), xon th (Spirochaeta), vi khun cc nh (Rickettsia, Mycoplasma, Chlamydia) v vi khun lam Cyanobacteria. Nhm sinh vt nhn thc (Eukaryote): bao gm vi nm(nm men, nm mc, nm nhy), ng vt nguyn sinh, vi to, thc vt, ng vt.
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V. VAI TR CA VI SINH VT TRONG I SNG

Cho n nay con ngi vn s dng cc phng php vi sinh vt c in nh sn xut ru, bia, men n bt m, cc sn phm sa nh vi khun lactic, dm nh vi khun acetic, ch bin tng, u nnh nh nm mc... Hu ht cc cht khng sinh u do vi sinh vt tng hp Mt s sn phm ca vi sinh vt cng c sn xut nh cc phng php vi sinh vt hin i: carotenoid v steroid t nm mc, acid glutamic v cc aminoacid khc cng nh nucleotid t vi khun. Nhiu enzym quan trng u do vi sinh vt tng hp. Ch c vi sinh vt mi c kh nng chuyn ha cc nguyn liu c bit, tr lng ln nh du la, kh t, cellulose thnh sinh khi hoc cc sn phm trung gian tit vo mi trng. K thut di truyn hin i c th a mt on DNA bt k vo vi khun, buc chng tng hp mt protein tng ng nh hormon, khng nguyn, khng th...Vic chuyn cc gene c nh N v cc gene khng su hi sang cy hay chuyn kh nng iu tr cc bnh do khuyt tt sinh ha gy nn, cng ang c quan tm c bit. Sau khi to DNA ti t hp, vic tip theo l a n vo li t bo cha mang plasmid c thc hin vi nhiu cch khc nhau:
1 2 3 Hnh 1.33: Mt s thit b chuyn DNA ti t hp vo t bo 1. My in bin np(Electroporation); 2. Vi tim(Microinjection); 3. Sng bn DNA (Biolistis apparatus) + Dung hp t bo trn (protoplast fusion): Dung hp 2 t bo loi b thnh t bo + Ha bin np: X l CaCl2 lnh, km sc nhit (420C trong 2 pht) + in bin np: c th n 109- 1010 th bin np/1gDNA. on bin np c kch thc 25-135kb (hnh 1.33.1) + Vi tim: Tim thng DNA ti t hp vo t bo (hnh 1.33.2) + Bn DNA vo t bo bng sng bn DNA (hnh 1.33.3) Ngoi ra cn nhiu phng php khc a DNA ti t hp vo t bo nh dng mng lipid (liposome) bao DNA a vo t bo hay phng thc di bom.
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Hinh 1.34 : Dung hp protoplast A. Cc protoplast; B. Hai protoplast dung hp trong mt cp; C. Cc protoplast c th dung hp trong th 3 (bn phi nh) hoc nhiu hn c khi ti 6 protoplast.
Hnh 1.35: Thao tc chuyn gen bng phng thc di bom Vit Nam, chng ta cng dng phng php trc tip bn gene v phng php gin tip chuyn gene bng con ng plasmid ch vaccine c kt qu. Thng 2 nm 2004, vin Pasteur thnh ph H Ch Minh cng gii m thnh cng b gene H5N1 (gy bnh cm ngi t g) c hng iu tr bnh ny. Nhng thnh tu khoa hc hin nay, nhng kinh nghim ca th gii chng minh rng vi sinh hc l ngnh khoa hc tuy mi pht trin nhng c ngha v mt l lun cng nh thc tin, c tng lai pht trin rc r trong th k XXI.
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VI. CC PHNG PHP V K THUT NGHIN CU

1. Phng php quan st t bo
Nh knh hin vi quang hc (knh hin vi thng) vi phng i 1500 - 2000 ln, c bit nh knh hin vi in t thng (TEM) v knh hin vi in t qut (SEM) m khoa hc c th thy c t bo, cu trc siu hin vi ca vi khun vi ng knh khong 1 m.
2 3 Hnh 1.36 : Mt s loi knh hin vi

1.Knh ca Leeuwenhoek; 2.Knh ca Robert Hooke, 3. Knh hin vi quang hc v 4. knh hin vi in t .
Hnh 1.37. Khun lc vi khun (tri); khun lc nm mc (gia) v khun lc nm men Quan st vi sinh vt trn tiu bn sng: vi sinh vt gia lam v lamella, nhum mc nho thy r mng nhy (capsule), y l phng php hay dng cho nhng vi sinh vt nui cy trn mi trng lng vi knh hin vi thng, quan st c kh nng vn ng ca chng.
Hnh 1.38: Vi khun c kch thc ln Thiomargarista namibiensis (tri)v siu vi khun Mycoplasma sp.
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Bng 1.6. Mt s phng php nhum mu v nguyn tc s dng Phng php nhum mu Nguyn tc s dng +Nhum n (xanh Dung dch ru hoc nc ca cc kim, dng quan st hnh methylene, carbolfuchsin, dng vi sinh vt, cch sp xp t bo crystal violet, safranin) +Nhum phn ly Vi cc phn ng khc nhau vi thuc nhum c th phn bit c chng - Gram Chia cc vi khun thnh 2 nhm ln: Gram dng gi mu tm ca crystal violet, Gram m mt mu khi ty do s nhum mu ph hng safranin Dng phn bit cc loi Mycobacterium v mt s loi Nocardia. Vi khun khng acid nhum vi carbolfuchsin v x l vi dung dch ru acid, vn gi mu . Vi khun khng khng acid s mt mu do s nhum mu ph l xanh methylene. Dng pht hin s c mt ca mng nhy, bi v polysaccharide mng nhy khng bt mu thuc nhum bao quanh t bo vi khun nhum mu
- Ziehl - Nielsen
+Nhum c bit -Nhum m (negative)
S dng pht hin bo t vi khun. khi dng thuc nhum lc malachite vi tiu bn c un nng, thuc nhum s thm -Nhum ni bo t nhp vo ni bo t v lm chng nhum mu lc, khi nhum b (endospore) sung bng safranin s lm phn bao quang bo t nhum mu hng. -Nhum tin mao Dng pht hin tin mao vi khun, s dng thuc lm (flagella) phng tin mao ri sau nhum bng carbolfuchsin - Quan st vi sinh vt trn tiu bn c nh v nhum mu: Phng php nhum Gram v Ziehl Nielsen... cho php nhn bit 2 nhm vi khun Gram dng va Gram m, hnh dng ca bo t, cc vt th n nhp nh ht d tr polyphosphate (ht d nhim sc, ht volutin), cc git m, glycogen Nhng tiu bn c nh ny c quan st bi gic ln X 90 hoc X 100 (bi gic dng du, vt knh c vng en). Tham gia vo c ch nhum mu c cu trc ca thnh t bo v bn cht cc hp cht ca sinh cht khc nhau hai loi vi khun. quan st nhng cu trc siu hin vi ngi ta dng knh hin vi in t TEM v SEM, c th thy c nhng cu to rt nh vi ln vi nanometre. Nhng vi khun Gram+ c mi lin h chng loi pht sinh gn gi nhau, y ngi ta chia thnh hai nhm ph: nhm c hm lng (G + X)% cao hn 50% nh cc Actinomycetales, Corynebacterium, Cellulomonas v nhm c (G + X)% thp hn 50% nh Clostridium, Bacillus, Staphylococcus, Nhng vi khun Gram- c mi quan h chng loi pht sinh cch xa nhau v y ngi ta phn ra rt nhiu nhm ph.
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Hnh 1.39. Cc bc tin hnh nhum Gram v v d minh ho kt qu.
(Quan st lam knh di knh hin vi: -Gram dng: xanh en hay tm -Gram m: vng hay ta. -Vi khun khng phn bit c vi phng php ny c gi l Gram bin i).
Hnh 1.40. So snh PG vi khun Gram dng (tri) v vi khun Gram m (phi)
2 3 Hnh 1.41. 1. Hans Christian Gram (1853-1938) 2 & 3. So snh PG vi khun Gram dng v vi khun Gram m
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Bng 1.7. Mt s tnh cht khc bit gia vi khun Gram dng v Gram m Tnh cht Gram dng Phn ng vi ha cht nhum gi mu crystal violet, do Gram t bo c mu tm hoc ta Lp peptidoglycan dy, nhiu lp Acid teichoic c Lp pha ngoi thnh khng c Lp lipopolysaccharide rt t hoc khng c Hm lng lipid v lipoprotein thp (vi khun acid c lp lipid mng lin kt vi peptidoglucan) Cu trc gc tin mao hai vng a gc To c t ch yu l ngoi c t (exotoxins) Chng chu vi tc nhn vt l kh nng chng chu cao Mn cm vi lysozyme rt mn cm, d b tan vi enzym ny Mn cm vi penicillin v sulfonamide Mn cm vi streptomycin, chloramphenicol, Tetracyclin Kt hp vi thuc nhum kim Mn cm vi cc cht ty anionic Chng chu vi mui Natri Chng chu vi kh hn cao thp cao, cht ch cao cao cao Gram m mt mu tm khi ty ra, nhum mu ph safranin hay Fuchsin mng, ch c mt lp khng c c nhiu, hm lng cao cao (to thnh lp ngoi thnh) bn vng a gc ch yu l ni c t (endotoxins) kh nng chng chu thp t mn cm (cn phi x l ph lp mng ngoi ca peptidoglycan) thp cao thp, lng lo thp thp thp
2. Phng php tch ly cc thnh phn ca t bo

Khi cn nghin cu cc thnh phn ring bit ca t bo, ngi ta phi tch ly cc thnh phn ny nh siu m, enzym lm tan thnh, cc khng sinh tc ng vo thnh, dng p sut thm thu gy co nguyn sinh, dng sc p c hc, dng siu li tm hoc li tm trong ng gradient Nh cc my o quang ph (Spectrophotometer) chng ta bit r rng v nhanh chng s lng t bo trong dch huyn ph. Nh phng php sc k giy hoc sc k ct chng ta c th thu c cc hp cht ring bit.
3.Phng php nghin cu tnh a dng ca VSV

3.1. Phng php nui cy - Nui tch ly (enrichment culture): dng mi trng chn lc v iu kin tt u lm tng t l hin din tng i ca vi sinh vt mc tiu t mt ngun t nhin
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- Nui tch ly = nui lm giu - Nui tch ly trong h kn: + Mi trng chn lc lng, iu kin ha l chn lc, ngun phn lp thch hp + Ch thu c cc qun th tng trng nhanh ca mt qun dng - Nhc im ca phng php nui tch ly trong phn tch tnh a dng: qun th thu c trong nui tch ly khng chc l qun th chim u th trong mu. 3.2. Phn lp v lm thun + Hp ria (streak plate): phn lp cc chng hiu kh hoc k kh khng nghim ngt + Pha long ti hn (extincting dilution): phn lp cc chng hiu kh + Pha long trong ng thch mm (agar shake tube method): phn lp cc chng k kh 3.3. Phng php nghin cu hot tnh vi sinh vt trong t nhin *Cc phng php truyn thng - Thng thng l cc phng php ex situ: mu c thu, bo qun v xc nh hot tnh ti phng th nghim. - o hot tnh tng ca vi sinh vt da trn mc tng trng: + Tng O2 tiu th, tng CO2 phng thch + Tng ATP - o tng hot tnh cc enzym ch yu trong cc chuyn ha vt cht do mt hoc mt s qun dng *Cc phng php in situ - Phng php in situ: o c hot tnh ca vi sinh vt ngay trong mi trng v vi mi trng - Phng php o in situ tng hot tnh da trn h hp, ATP, o hot tnh chuyn ha - Phng php ng v phng xa: o hot tnh chuyn ha vi nhy cao + Hot tnh ring (specific activity): t l hot tnh trn c cht ng v so vi hot tnh tng + Loi b hot tnh do phn ng phi sinh vt: i chng t bo cht (killed cell control) - Phng php vi in cc (microelectrode): theo di bin i v thnh phn ha hc trong vi mi trng + Cc vi in cc pH, oxygen, N2O, CO2, H2, H2S + Di chuyn theo cc bc 1mm bng micromanipulator + Dng nghin cu cc chuyn ha trong cc li sinh khi (micromat) ng dng FISH nghin cu hot tnh ca qun dng trong k thut ISRT - ISRT (In situ Reverse transcriptase): + Lai in situ mRNA bng primer chuyn bit + Thc hin phin m ngc to cDNA v khuch i bng PCR + Lai vi mu d nh du hunh quang - ng dng: + Nghin cu xc nh vi sinh vt c biu hin gen trong qun x mt thi im nht nh + Xc nh qun th c hot tnh trong qun dng v qun x + Nghin cu nh hng ca cc yu t mi trng ln s biu hin ca gen trong qun x.
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Cu hi n tp chng 1
1. Nhng sinh vt no c gi l vi sinh vt ? Vi sinh vt khc vi vt v sinh bi nhng c im no? 2. Nhng im ging nhau v khc nhau gia t bo nhn s v t bo nhn thc. 3. V tr ca vi khun, x khun, nm mc, nm men trong cc h thng phn loi hin nay ? 4. T bo...........khng c nhn thc nhng c ............phn tn trong t bo cht, ni chung ch c 1......, l t bo....... 5. Nguyn tc phng php nhum Gram, vi sao khi nhum theo phng php Gram th vi khun Gram dng bt mu tm ? 6. Vai tr ca vi sinh vt trong thin nhin v trong i sng con ngi ?
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc; NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty. Vi sinh vt hc, 1997, NXBGD H Ni.
* Ti liu tham kho

1. Nguyn Thnh t . 1999. Vi sinh vt hc, NXBGD, H Ni. 2. http.//vietsciences.net v http.//vietsciences.free.fr 3.Brock, 2006. Biology of Microbiology
* Gii thch thut ng

Loi : Theo ngha rng, loi l mt nhm cc c th sinh vt m c nhng c im sinh hc tng i ging nhau v c kh nng giao phi vi nhau cho con ci hu th. Cn theo nh ngha ca Ernst Mayr, loi l nhm cc qun th t nhin c kh nng giao phi vi nhau v tng i cch ly sinh sn vi cc nhm khc. Chi (genus) mt n v phn loi sinh hc dng ch mt hoc mt nhm loi c kiu hnh tng t v mi quan h tin ha gn gi vi nhau. H (Familia) L tp hp ca cc CHI gn nhau nht. H c th chia nh thnh cc phn h, cc tc, cc phn tc. Tn h cc vi sinh vt thng c tn cng l ceae, cn cc loi ng vt l idae. B (Order): Bc phn loi trn H trong h thng thang bc phn loi sinh vt. L tp hp ca cc h gn nhau nht. Tn B ca cc vi sinh vt c phn ui l les.
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Lp (Class): Tp hp nhng B c quan h gn nht. Ngnh (Phylum) Bc phn loi trn LP trong h thng thang bc phn loi sinh vt. L tp hp ca cc lp gn nhau nht. nhng ngnh c c im chung nht c xp vo mt gii. Gii (Kingdom): S phn chia cc gii c s thay i rt ln trong lch s pht trin ca sinh hc. u tin Linnaeus (1735) chia th gii ny thnh hai gii ng vt v Thc vt. Haeckel (1866) chia thnh ba gii gii ng vt, Thc vt v Sinh vt nguyn sinh. Sinh vt nguyn sinh (protista) bao gm cc sinh vt cc sinh vt c c im chung l c th c mt t bo nhn thc (ng vt nguyn sinh v to n bo). Nm 1969 Whittaker chia gii sinh vt ra lm nm gii:
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- Ribosome: 70S (Bacteria v Archaea), 80S (Eukarya) - Aminoacid u tin: formylmethionine (Bacteria), methionine (Archaea v Eukarya) - Tc nhn c ch sinh tng hp protein vi khun khc vi Archaea v Eukarya Phn loi hc truyn thng v kha phn loi Bergey - Phn loi hc vi khun: sp xp cc loi vi khun da trn cc c im kiu hnh - Cc b su tp ging thng lu gi cc chng in hnh (type starin) dng lm chng chun c cc c im ca loi - Cc c im kiu hnh quan trng: nhum Gram, hnh thi t bo v s hin din ca cc cu trc t bo nh ni bo t, phn ng sinh ha, phn ng min dch - Kha phn loi Bergey (Bergey's Manual of Systematic Bacteriology) gm cc c tnh kiu hnh dng sp xp vi khun theo phn loi truyn thng - Cc bc phn loi: + Dng (clone): cc t bo ca cng mt qun th + Chng (strain): cc chng kh ng nht v mt di truyn + Loi: (species) cc chng c c im kiu hnh, kiu gen ging nhau + Ging (genus), h (family), b (order), lp (class), ngnh (Phylum), gii (kingdom), lnh gii (domain).
Chng 2: Sinh hc ca cc c th v bo
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CHNG 2
SINH HC CA CC C TH V BO
I. KHI NIM V VIRUS, VIRION, VIROID V PRION
1. Virus l g?
Virus l mt dng sng n gin nht hin nay ch cha mt loi nucleic acid nn h gen ca chng l DNA hoc RNA, sng k sinh bt buc trong t bo ch c gi l ht virus hay virion. Kch thc nh. Virus c kch thc rt nh t 10nm n 300nm trong khi kch thc ca vi khun khong 1000nm v kch thc ca hng cu l 7500nm. V vy, virus ch c th quan st c trn knh hin vi in t.
T bo V Nhn
Vi khun
Hnh 2.1. Kch thc ca virus so vi vi khun, nhn v t bo ng vt
Genome virus ch cha mt loi nucleic acid, c th l DNA hoc RNA, c th dng thng hoc khp kn, chui n hoc chui kp. Genome phn on hoc khng phn on. L dng sng khng c hot tnh trao i cht. Virus khng c ribosome hot ng hoc khng c b my tng hp protein. Cho nn mc d mt s virus c enzym ring cu mnh nhng virus ch c th nhn ln trong t bo sng, iu khin b my sinh tng hp ca t bo ch phc v cho mnh to thnh cc ht virus mi. Ch "virus" c xut x cng t ch virus trong ting Latinh ch cht c, ting Hn ngy nay gi n l bnh c . Ngy nay bn cnh ngha sinh hc nh trn, "virus" cn c dng theo ngha n d ch cc nhng g sinh sn k sinh, nh virus my tnh. Tng ht virus c gi l virion hay viron.
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Hnh 2.2. Ba loi virus thng gp: virus ca vi khun, cn gi l thc khun th (tri); virus ca ng vt (phi trn); v retrovirus (virus phin m ngc, phi di)
2. c im chung ca virus
Virus l mt kt cu i phn t v bo, khng c h thng sinh sn nng lng, khng c ribosome, khng sinh trng c th, khng phn ct v khng mn cm vi cc cht khng sinh, ch cha 1 trong 2 loi nucleic acid (DNA hoc RNA) C s giao t tng h gia trng thi k sinh trong t bo vt ch v trng thi phi sinh vt (trng thi khng sng). Khi ngoi t nhin chng tn ti th virion, khng trao i cht, khng cm ng, khng sinh sn. Kch thc nh (0,02 0,03mm), ch sao chp trong t bo ch Chu lnh tt, chu nhit v ha cht km. Khng nui cy c trn mi trng thch thng. T bo vi khun, ng vt, thc vt u c th b nhim bi cc virus chuyn bit
3. Virion, viroid, prion

3.1. Virion Virion l cc ht virus c lp rp bn trong t bo t cc cu phn c bit, mang b gene virus. Cc virion khng t pht trin hay phn chia v c coi nh giai on ngoi t bo ca virus (extracellular phage). C th hnh dung cc virion nh nhng con tu v tr mang b gene virus t t bo ny sang t bo khc ng thi bo v b gene virus trong mt mi trng "khng thun tin" m virus khng th nhn ln c. Tnh cht c bn ca virion c th hin bng sau:
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Bng 2.1: So snh virus, viroid, prion v virion Tnh cht Virus Viroid Prion C cu to t bo Ch cha DNA hoc RNA Cha c DNA v RNA Ch cha RNA Ch cha protein Cha ribosome Sinh sn c lp
Virion
3.2. Viroid: L nhng phn t RNA kn, c tnh cm nhim c 150 ribonucleotit, dng trn khng c v capsid, mch n c pht hin nm 1971, l tc nhn gy bnh nh nht m con ngi bit (Bnh c khoai ty hnh thoi, bnh ln thc vt, bnh hi cy da...). Viroid thm ch khng m ha bt k mt protein no v s nhn ln ca chng ph thuc hon ton vo s hot ng cu enzym t bo ch
Hnh 2.3. Phn t viroid v c khoai ty b bnh c dng hnh thoi 3.3. Prion: Ch cha thnh phn protein v khng cha mt loi acid nucleic no. Trong c th bnh thng c th c sn cc prion nhng chng khng gy bnh. Trong iu kin no prion c th thay i cu trc v gy bnh gy bnh. Prion gy nhiu bnh nguy him ng vt v ngi, gy thoi ha h thn kinh trung ng v gim st tr tu nh bnh BSE (xp no b in)...
Stanley B. Prusiner (1942- ) Prion B b bnh BSE Nm 1981, Stanley B. Prusiner(1942- ) chng minh nguyn nhn gy ra cn bnh trn c bn cht l protein v c gi l prion, c ngha l protein gy nhim (proteinaceous infection particle). ng l ngi u tin phn lp c prion v nh cng trnh ny ng nhn gii Nobel Y hc nm 1997.
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Bng 2.2 . So snh gia virus v prion c im Virus Prion Qua c mng lc vi khun C C Khng hoc rt t (~50 nucleotid khng m ha cho bt k protein Cha acid nucleic C no ca prion) C hnh dng c trng di C Khng knh hin vi in t Cha protein C C B bt hot bi: + Formaldehyde C Khng +Protease t Khng 0 +Nhit (80 C) Phn ln Khng +Ion ha v tia UV t Khng Bnh l: +Thi gian bnh Ty thuc loi virus Di +Phn ng min dch C Khng +To interferon C Khng +Gy phn ng vim C Khng Bng 2.3. Cc bnh do prion gy ra ngi TT 1 2 3 4 5... Tn bnh Creutzfeldt- Jacob (CJD) Mt ng gy cht, di truyn theo gia nh (FFI- Fatal Familial insomnia) Gerstmann Straussler-cheinker (GSS) Kuru Alpers Dng bnh Bnh ri rc ly nhim bnh vin hoc trong phm vi gia nh Phm vi gia nh Phm vi gia nh Truyn nhim Xy ra tr s sinh Bng 2.4. Cc bnh do prion gy ra ng vt TT 1 2 3 4 5 6 7 8... Mo Gia sc ln Bo Linh dng chu Phi Sn dng (Gemsbok) Hu, nai Cu Vt ch Sn dng rp Bnh xp no Xp no kiu di Xp no (b in) Xp no kiu di Xp no Xp no Bnh no mn tnh (Chrohic wasting disease- CWD) Bnh cu gii (scrapie) Bnh
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3.3.1. Khi qut v prion Protein prion vit tt l PrP l sn phm bnh thng ca gen protein prion, gi tt l PRNP (normal product of prion gene) c biu hin hu ht cc ng vt c v, c bit c biu hin mc cao trong cc t bo m no. C hai loi PrP: loi bnh thng khng gy bnh ca t bo c k hiu l PrPc (c-cell) v loi gy bnh c k hiu l PrPSc (sc-scrapie). PrPSc c khi lng phn t 27 000 30 000. PrPc c cu trc bc hai ch yu l dng xon (thng c 3 chui nh vy) cn PrPsc ch yu c cu trc phin gp np . PrPc c th gy nhim t c th ny sang c th khc. Khi trong c th n c th bin cu trc bc hai PrPc PrPSc v gy bnh. Cc t bin lm thay i cu hnh PrPc lm cho n ngu nhin tr thnh PrPSc . c bit mt s c hnh thnh s bin i PrPc xung quanh thnh PrPSc. C trng hp bin i PrPc thnh PrPSc mt cch tnh c m khng cn c t bin. iu ny c th l gii v sao bnh prion c th l do ly nhim, do di truyn hay xut hin mt cch tnh c.
Hnh 2.4. Hai dng cu trc ca prion Prion c th chui qua mng lc c 0,1 m (mng lc vi khun l 2,2 m), ch b bt hot vi dung dch NaOH 1M 550C hay cht ty ra 20000ppm(nng bt git gia nh thng l 5000ppm). Bng 2.5. So snh PrPc v PrPSc c im Cu trc Khng protease C mt trong si Scrapie Nm trong hoc trn t bo Thi gian quay vng (chu k) C C Trong bng sinh cht Nhiu ngy PrPc Dng cu Khng Khng Trn mng sinh cht Nhiu gi PrPSc Dng ko di
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Hnh 2.5. Prion v c ch gy bnh BSE (xp no b in) 3.3.2. Cu trc ca prion chui PrPc l mt protein gm 4 chui k hiu l H1, H2, H3 v H4 cha khong 40% v 45% di np gp .
Vng u NH2 ca PrPc to ra b mt chung ni vi PrPSc. u NH2 rt linh ng, cu trc bc ba ca u NH2 lin quan n s thay i cu trc trong qu trnh hnh thnh PrPSc. Vng u COOH ca PrPc l v tr gn vi protein. PrPSc c to thnh do s tho xon u NH2 ca PrPc to thnh di gp np .Gii gp np ny nhiu (45%) nn cu trc n nh hn so vi PrPc, ngn cn s phn gii ca protein. PrPSc gy bnh dng dimer. 3.3.3. S nhn ln ca prion Trong cc t bo khng b nhim PrPc vi trnh t kiu di WT (wild typ) tn ti trong trng thi cn bng dng monomer xon , mn cm vi protease hoc gn vi protein X (Hnh 2.6). Protein X PrPc PrPsc PrP*/Protein X PrPsc /PrP*/Protein X (PrPSc)2 + Protein X
Hnh 2.6. S cho thy s to thnh PrPc nh khun c sn Bc u PrPc kt hp vi Protein X to thnh phc hp PrP*/Protein. Tip n l PrPc lin kt PrP trong phc hp PrP*/Protein. Khi PrP* c chuyn dng thnh phn t mi, protein X s ri khi phc h, ch cn li dng dimer (hoc oligome). Trong c th bnh thng, prionc to thnh bi t bo thn kinh v mt s t bo
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nht nh. Vic to ra PrPc bt u t nhn, mRNA dng dch m cho PrPc c to thnh trong nhn, xuyn qua l mng nhn ra sinh cht gn vo ribosome to phc nh vo mng li ni cht ht (RER- rough endoplasmid reticulum), sau PrPc c tng hp v vn chuyn thng qua b my Golgi. pha nh b my ny cc bng (kiu khngbo) cha PrPc ny chi v vn chuyn n b mt t bo. y chng dung hp vi mng sinh cht v phn t hnh que chui ra ngoi t bo. Prion c th i vo no dc theo si trc (axon) ca t bo thn kinh. Prion c th nhn ln trong t bo lympho do di chuyn c trong mu. Vhng c th nhn ln trong cc t bo sao (aster) v cc t bo thn kinh m khc. Sau khi n phi thc n nhim prion, chng c t bo lympho hp th ri i vo cc m bch huyt, mng payer, amidan. Ti y c cc dy thn kinh qua prion c th xm nhp vo h thng thn kinh trung ng. 3.3.4. Nghin cu bin php chng bnh prion Khoa hc hin nay ang a ra cc phng n thit k cc loi thuc nhm ngn cn s to thnh prion gy bnh t prion bnh thng: -To dc phm c cu trc ging nh protein X cnh tranh gn vo PrPc lm n nh cu trc ca PrPc v khng cho n chuyn ha thnh PrPSc. -Tm khng th gn vo PrPc. -To ra cc cht ha hc c th chuyn PrPc thnh PrPSc. Mt s cht c hot tnh ny nh dimethylsunfocid, oxid trimethylamin N, mt s polyol khc v mt s loi ng. -Ngn cn s tho thnh PrPSc nh acid nucleic. -S dng cc hp cht, v d cht dn xut ca 1,2-hydroquinon s bo v t bo khi b ph hy. -To ra cc ng vt chuyn gen khng mang gen PrP, do c th khng c bnh prion.
4. Lch s pht hin virus

Ngay t nm 1883 nh khoa hc ngi c Adolf Mayer (1843-1942) khi nghin cu bnh khm cy thuc l nhn thy bnh ny c th ly nu phun dch p l cy b bnh sang cy lnh, tuy nhin ng khng pht hin c tc nhn gy bnh. Nm 1884 Charles Chamberland (1851-1908 ) sng ch ra mng lc bng s tch cc vi khun nh nht (Hnh 2.3). Nm 1892 nh thc vt hc ngi Nga Dmitri Iwanowski (1864-1920) dng mng lc ny nghin cu bnh khm thuc l. ng nhn thy dch p l cy b bnh cho qua mng lc vn c kh nng nhim bnh cho cy lnh v cho rng tc nhn gy bnh c l l vi khun c kch thc nh b n mc c th i qua mng lc, hoc c th l c t do vi khun tit ra.
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2 3 Hnh 2.7: 1. Adolf Mayer (1843-1942); 2. Mng lc bng s; 3. Charles Chamberland (1851-1908 ) Gi thuyt v c t qua mng lc b bc b vo nm 1898 khi nh khoa hc ngi H Lan Martinus Beijerinck (1851-1931) chng minh c rng tc nhn ly nhim l cht c sng (Contagium vivum fluidum) v c th nhn ln c. ng tin hnh phun dch p l cy bnh cho qua lc ri phun ln cy v khi cy b bnh li ly dch p cho qua lc phun vo cc cy khc. Qua nhiu ln phun u gy c bnh cho cy. iu chng t tc nhn gy bnh phi nhn ln c v nu l c t th nng lc gy bnh s phi dn mt i. Nm 1901 Walter Reed v cng s Cuba pht hin tc nhn gy bnh st vng, cng qua lc. Tip sau cc nh khoa hc khc pht hin ra tc nhn gy bnh di v u ma. Tc nhn gy bnh u ma c kch thc ln, khng d qua mng lc, do cc tc nhn gy bnh ch n gin gi l virus. Nm 1915 nh vi khun hc ngi Anh Frederick Twort (1877-1950) v nm 1917 nh khoa hc ngi Php Felix d'Hrelle (1873-1949) pht hin ra virus ca vi khun v t tn l Bacteriophage gi tt l phage. Nm 1935 nh khoa hc ngi M Wendell Stanley (1904-1971) kt tinh c cc ht virus gy bnh m thuc l (TMV). Ri sau TMV v nhiu loi virus khc u c th quan st c di knh hin vi in t.Nh vy nh c k thut mng lc em li khi nim ban u v virus v sau nh c knh hin vi in t c th quan st c hnh dng ca virus, tm hiu c bn cht v chc nng ca chng. Ngy nay virus c coi l thc th cha c cu to t bo, c kch thc siu nh v c cu to rt n gin, ch gm mt loi acid nucleic, c bao bi v protein. Mun nhn ln virus phi nh b my tng hp ca t bo, v th chng l k sinh ni bo bt buc. Virus c kh nng gy bnh mi c th sng t vi khun n con ngi, l th phm gy thit hi nng n cho ngnh chn nui, gy tht bt ma mng v cn tr i vi ngnh cng nghip vi sinh vt. T nhng thp k cui ca th k XX tr li y ngy cng xut hin cc dng virus mi l ngi, ng vt m trc y hc cha h bit ti, e do mng sng ca con ngi. Sau HIV, SARS, Ebola, cm A H5N1 s cn bao nhiu loi na s xut hin gy tai ho cho con ngi.
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Mt khc, do c cu to n gin v c genom nhiu kiu vi c ch sao chp khc hn cc c th khc nn virus c chn l m hnh l tng nghin cu nhiu c ch sinh hc mc phn t dn n cuc cch mng sinh hc cn i: Sinh hc phn t, di truyn hc phn t. V nhng l do trn vic nghin cu virus c y mnh v tr thnh mt ngnh khoa hc c lp rt pht trin.
II. HNH THI V CU TRC CA VIRUS

1. Hnh thi
1.1. Cu to c bn Tt c cc virus u c cu to gm hai thnh phn c bn: li l acid nucleic (tc genom) v v l protein gi l capsid, bao bc bn ngoi bo v acid nucleic. Phc hp bao gm acid nucleic v v capsid gi l nucleocapsid hay xt v thnh phn ho hc th gi l nucleoprotein. i vi virus RNA th cn gi l ribonucleoprotein Genom ca virus c th l DNA hoc RNA, chui n hoc chui kp, trong khi genom ca t bo lun l DNA chui kp, v trong t bo lun cha hai loi acid nucleic, DNA v RNA.
Dmitri Iwanowski (1864-1920)
Martinus Beijerinck (1851-1931)
Walter Reed (1851-1902)
Felix d'Herelle (1873-1949)
Frederick Twort (1877-1950)
Wendel M.Stanley (1904-1971)
Hnh 2.8. Mt s nh khoa hc nghin cu virus 1.2. V capsid Capsid l v protein c cu to bi cc n v hnh thi gi l capsome. Capsome li c cu to t 5 hoc 6 n v cu trc gi l protome. Protome c th l monome (ch c
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mt phn t protein) hoc polyme (c nhiu phn t protein).
Hnh 2.9. Hnh thi mt s loi virus i xng xon: virus khm thuc l; i xng hnh khi: Adenovirus v cu trc phc tp: Phage T4 - Pentame (penton) c 5 protome nm trn cc nh ca khi a din, cn hexame (hexon) to thnh cc cnh v b mt hnh tam gic. Capsid c kh nng chu nhit, pH v cc yu t ngoi cnh nn c chc nng bo v li acid nucleic - Trn mt capsid cha cc th th c hiu, hay l cc gai glicoprotein, gip cho virus bm vo cc th th trn b mt t bo. y cng chnh l cc khng nguyn (KN) kch thch c th to p ng min dch (MD). - V capsid c kch thc v cch sp xp khc nhau khin cho virus c hnh dng khc nhau. C th chia ra ba loi cu trc: i xng xon, i xng hnh khi v cu trc phc tp. Cu to chung ca virus gm 2 phn chnh Li( b gen): acid nucleic V (capsid): protein Phc hp gm acid nucleic v protein c gi l Nucleocapsid
Li (b gen) acid nucleic
Acid nucleic Nucleocapsid
V (capsid) protein
Capsid
Hnh 2.10. Cu to chung ca virus
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Hnh 2.11. So snh b gen ca virus vi b gen ca sinh vt nhn thc
2. Kch thc ca virus

Tuyt i a s virus c kch thc rt nh. Ngi ta thng o kch thc virus bng n v nanometre (nm, 1nm= 10-9m). Vi knh hin vi in t v cc k thut ph tr, ngy nay ta c th o c, quan st t m hnh thi tng loi virus. V d: Virus RNA Picoviridae c kch thc c 20-30nm, Retroviridae-100-120nm Virus DNA: Parvoviridae c kch thc 18-26, Poxviridae-130-300nm.
3. Cu trc ca virus
Virus c cu to rt n gin, bao gm li l acid nucleic, tc genome nm pha trong cn pha ngoi c bao bc bi v protein, v protein bo v genome khi s tc ng ca cc yu t mi trng v d nh nuclease trong mu. V protein c gi l capsid. Capsid c cu to bi cc n v hnh thi l capsome. Capsome li c cu to bi cc n v cu trc l protome. Protome c th l monome (ch c mt phn t protein) hoc polyme (nhiu phn t protein). Capsid v acid nucleic c gi l nucleocapsid. Li l acid nucleic, v l capsome l protein, hp li thnh nucleocapsid. Nucleocapsid c bao bc bi lp v ngoi (lipoprotein) vi cc gai. Mt s virus cn cha v ngoi, bao bc bn ngoi capsid. V ngoi c bn cht l lipoprotein cha khng nguyn ca virus. V ngoi mt phn bt ngun t mng sinh cht ca t bo ch khi virus chui ra ngoi theo li ny chi. mt s virus, v ngoi c ngun gc t mng nhn ca t bo. Ht virus nguyn vn cn c gi l virion.
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Hnh 2.12. Cu trc c bn ca virion

A.S virus a din n gin nht, mi mt hnh a din l tam gic u. nh do 5 cnh hp thnh. Mi cnh cha 3 capsomer B. S ca virus hnh que vi cu trc i xng xon (virus khm thuc l). Capsomer sp xp xon xung quanh si acid nucleic dng xon c.
Virus c 3 kiu cu trc 1. Hnh tr i xng xon
2. Hnh khi
3.Dng phi hp
Khi a din
Khi cu
Virus khm thuc l
Virus bi lit
Virus HIV
Phage T2
Hnh 2.13. Cc kiu cu trc i xng virus Cu trc hnh khi. Capsid c cu trc hnh khi a din virus bi lit, hnh khi cu virus HIV. Cu trc xon. Nucleocapsid dng ko di. Cc capsome sp xp xung quanh theo chiu xon ca acid nucleic. a s virus c cu trc xon c v ngoi bao bc nucleocapsid xon. Cu trc phc tp. Cu trc hn hp va dng khi va dng xon. V d phage c u dng khi, ui dng xon trng nh con nng nc.
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4. Genom ca virus
Genom ca virus rt a dng v cu trc, kch thc v thnh phn nucleotid. Chng c th l DNA hoc RNA, chui n hoc kp, thng hoc khp vng. Bng 2.6. Cc dng genom virus
Loi acid nucleic DNA n Cu trc Chui n, dng thng Chui n, khp vng V d Virus parvo Phage X174, M13, fd Herpes, adeno, coliphage T, phage l.
DNA kp
RNA n
RNA kp
Chui kp, dng thng Coliphage T5 Chui kp, dng thng, trn mt mch Vaccinia, Smallpox c nhng ch t cu ni Polioma (SV40), papiloma, phage PM2, phosphodieste. virus m hoa l Chui kp vi hai u khp kn Chui kp khp vng kn Chui n, dng dng thng Picorna (polio, rhino), toga, phage RNA, Chui n, m, dng thng MTV v hu ht virus thc vt. Chui n, dng, dng thng, nhiu Rhabdo, paramyxo, (si, quai b) on. Virus m cy tc mch (Bromus) (cc Chui n, dng dng thng gm hai on c bao gi trong cc virion tch bit). on gn vi nhau. Retro (HIV, Sarcoma Rous) Chui n, m dng thng, phn on Orthomyxo (cm) Reo (rota), mt s virus gy u thc vt, Chui kp, dng thng, phn on NPV cn trng, phage j6 v nhiu virus nm (mycovirus).
Kch thc genom c th t 3500 nucleotid ( phage nh) n 560.000 nucleotid ( virus herpes). Cc trnh t genom virus phi c c m bi t bo ch, cho nn cc tn hiu iu khin phi c cc yu t ca t bo ch nhn bit. Cc yu t ny thng lin kt vi protein virus. Do c kch thc nh nn genom virus tin ho s dng ti a tim nng m ha ca mnh. V th hin tng gen chng lp v hin tng ct ni (splicing) mRNA virus l rt ph bin. Kch thc genom thay i rt nhiu cc virus khc nhau. Cc genom nh nht (v d Bactariophage MS2, Q ) c kch thc 1x106 Da m ha cho 3-4 protein. Mt s virus khc tn dng ti a khng gian ca genom bng cch s dng cc gen chng lp, tc l cc gen gi ln nhau trn cng khung c, ch khc nhau dim khi u hoc kt thc. Cc genom ca coliphage T chn, herpes, vaccinia c kch thc 1,6x108 Dal c th m ha cho 100 protein. Mt s c im ca genom virus cn lu : - Genom DNA kp (v d virus pox, herpes v adeno) thng c kch thc ln nht. - Genom DNA kp khp vng (siu xon hoc khng siu xon) thng thy phage Genom DNA kp virus vaccinia c hai u khp kn
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DNA n dng thng (v d virus parvo) c kch thc rt nh. - Cc DNA dng thng thng c trnh t lp li u. - Tt c genom RNA kp u phn on (cha mt s on khng ging nhau, mang thng tin di truyn tch bit). - Genom RNA n c phn thnh RNA dng (genom +) v RNA m (genom -) da vo trnh t nucleotid ca mRNA. - Phn ln genom RNA n u khng phn on tr virus orthomyxo (virus cm). Virus retro c genom l hai phn t RNA n ging nhau, ni vi nhau u 5 nh cu ni hydro. - Virus m cy Alfalfa (AMV) c genom gm 4 on RNA n, dng, dng thng, c gi vo 4 v capsid khc nhau nn cn gi l virus d capsid (heterocapsidic) phn bit vi virus m tt c cc on u c gi trong mt htvirus ng capsid (isocapsidic)
III. NUI CY VIRUS

Do virus ch sinh sn bn trong t bo sng nn phi c cc phng php c bit nui cy chng. C 3 h thng chnh dng nui cy virus trong phng th nghim.
Hnh 2.14. Vt ch ca virus trong phng th nghim - Nui cy m t bo. Cc t bo c ngun gc t cc m ca ngi hay ng vt c nui trong bnh cha mi trng nhn to, cho pht trin v dng lm ngun nguyn liu cy virus. - Phi g. Mt s virus c th nhn ln trong t bo phi g 6- 13 ngy. Ngy nay phng php nui ny c thay th bi t bo nui cy m. Tuy nhin trong sn xut mt s loi vaccine, phng php ny vn c s dng. - ng vt thc nghim. Trc y phng php ny c dng ph bin phn lp v nghin cu virus. Cc ng vt c s dng l chut, th, kh, chn...Tim hn dch nghi l c virus vo ng vt v quan st bnh cnh lm sng. Hin nay phng php ny vn c dng phn lp mt s virus.
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Hnh 2.14b. nh lng virus bng k thut plaque (vng tan)
IV. NH HNG CA VIRUS LN T BO

Virus c th tc ng ln t bo theo 4 cch sau: - Gy cht t bo. Kt qu ca vic nhim virus l lm cho t bo b hu hoi, dn n lm cht t bo (CPE- Cytopathic effect). - Chuyn dng. T bo b nhim virus nhng khng cht m chuyn t trng thi bnh thng sang trng thi c bit, thnh cc t bo u hoc ung th. - Nhim tim tng. Virus tn ti bn trong t bo trng thi hot ng tim n nhng khng nh hng r rt n chc nng ca t bo. - Gy ngng kt hng cu. Mt s virus trn b mt v ngoi c cha protein gy ngng kt hng cu (Haemaglutinin) gn trn b mt cc t bo nhim. Khi thm hng cu vo th hng cu s b kt dnh bi cc t bo nhim.
V. CC BNH DO VIRUS
Virus l tc nhn gy bnh quan trng cho ngi, ng vt, cy trng v vi sinh vt. a s cc bnh thng gp ngi l do virus. Hu ht chng gy bnh th nh, bnh nhn t bnh phc sau mt thi gian nht nh. Nhiu loi tn ti thm lng trong c th. Chng nhn ln nhng khng gy bt k triu chng no. Tuy nhin vic nhim virus thng th nh, nhng i khi c th gy bnh trm trng nhng ngi mn cm bt thng. Mt s do virus gy bnh rt nng v thng c t l t vong cao. Sau y l mt s virus gy bnh.
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Bng 2.7. Virus gy bnh cha genome DNA

Nhm virus Pox Herpes Tn virus Variola Molluscum (u mm) Herpes simplex Varicella zoster Cytomegalo EB ( Epstein- Barr), HH6 Virus adeno Vim gan B Papiloma Virus JC B19 Tn bnh u ma U mm ly Herpes Thu u zona (shingles) Nhim trong tho hip min dch Bnh bch cu n nhn ly nhim Bnh ngoi ban t ngt Vim hng Vim kt mc Vim gan Mn cc Vim cht trng no nhiu tin trin Ban truyn nhim, cn bt sn
Adeno Hepadna Papova Parvo
Bng 2.8. Virus gy bnh cha genome RNA

Nhm virus Orthomyxo Paramyxo Tn virus Virus cm cm;Hp bo h hp Si Quai b Virus corona Virus di Entero Rhino Vim gan A SRSV (virus c cu trc dng trn nh- small round structure virus) Alpha (virus arbo nhm A) Rubi Flavi (virus arbo nhm B) Vim gan C Mt s virus arbo Tn bnh Cm Vim nhim ng h hp Si Quai b Gy nhim ng h hp SARS Bnh di Vim no, bi lit Cm lnh Vim gan Vim d dy, rut
Corona Rhabdo Picorna
Calici
Toga Flavi
Bunya
Reo Arena
Retro
Rota Vim mng no m ri mng mch lympho bo Virus Machupo;Virus Junin Virus lassa HTLV-I, II HIV- 1, 2 Virus Marburg Virus E bola
Vim no St xut huyt Rubeon (si c) Vim no St xut huyt Vim gan Vim no St xut huyt St, vim thn Bnh ng tiu ho Vim mng no St xut huyt Ung th t bo T U lympho; Lit AIDS St Marburg St xut huyt Ebola
Filo
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VI. NH HNG CA TC NHN VT L, HA HC N VIRUS

- Nhit cao: a s virus b bt hot 560C trong vng 30 pht, hoc 1000C trong vi giy. - Nht thp: a s virus bn nht lnh nn c th bo qun lu ( 700C). Mt s virus b bt hot trong qu trnh lm ng lnh hoc tan bng. - Kh hn: Kh nng chu kh hn ca virus khc nhau tu loi. Mt s sng st, mt s b bt hot nhanh iu kin kh hn - Bc x t ngoi: Virus b bt hot bi tia t ngoi - Chlorofoc, ethe v cc dung mi khc: Cc virus c v ngoi cha lipid s b bt hot, cn khng cha lipid s bn vng. - Cc cht oxi ha v cht kh. Virus b bt hot bi di tc dng ca formaldehyd, clo, iod v H2O2. - propiolacton v formaldehyd l cc ho cht c dng bt hot virus trong sn xut vaccine, song a s virus khng b bt hot bi phenol. - Cht kh trng virus: Tt nht l dng dung dch hypoclorua (mt cht n mn) v glutaraldehyd (l cht c th gy mn cm v kch thch gy kh chu chy nc mt cho ngi dng).
VII. CON NG LY NHIM VIRUS VO C TH

Virus vo c th theo 4 con ng chnh: - Ht th: Qua ng h hp - n ung: Qua ng tiu ho (d dy- rut) - Xm nhp qua da, vt xc nim mc (qua quan h tnh dc), truyn mu, tim chch, phu thut cy ghp hay do cn trng hoc ng vt cn. - Bm sinh: Do m truyn qua nhau thai sang con C ch gy bnh ch yu ca virus l s xm nhp. Bnh sinh ra do virus lan truyn trc tip ti cc m v c quan trong c th. S sinh sn ca virus trong t bo s git cht t bo (tuy nhin c trng hp khng git t bo). Tc ng gy hu hoi t bo c gi l CPE (cytopathic effect) s dn n tn thng v hu hoi chc nng ca cc m v c quan, ri t biu hin ra cc du hiu, triu chng.
VIII. CC QU TRNH NHN LN CA VIRUS

Virus khng c hot tnh trao i cht m s dng b my trao i cht ca t bo tng hp cc thnh phn thit yu ca mnh, sau lp rp to ra cc ht virus con ging nh nguyn bn. V vy ngi ta thng s dng thut ng nhn ln (nhn bn) ca virus thay cho t sinh sn.
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S nhn ln ca virus c th lm tan t bo gi l chu trnh sinh tan hoc c th gn genome ca mnh vo nhim sc th ca t bo, tn ti lu di di dng tim n m khng l cht t bo gi l chu trnh tim tan. Virus khng c hot tnh trao i cht m s dng b my trao i cht ca t bo tng hp cc thnh phn thit yu ca mnh, sau lp rp to ra cc ht virus con ging nh nguyn bn. V vy ngi ta thng s dng thut ng nhn ln (nhn bn) ca virus thay cho t sinh sn. S nhn ln ca virus c th lm tan t bo gi l chu trnh sinh tan hoc c th gn genome ca mnh vo nhim sc th ca t bo, tn ti lu di di dng tim n m khng l cht t bo gi l chu trnh tim tan.
Hnh 2.15. Cc phng thc xm nhp ca virus vo t bo 1. Virus c v ngoi: a) Dung hp vi mng sinh cht, y nucleocapsid vo t bo. V ngoi virus nm trn mng sinh cht (v d virus paramyxo, herpes). b) Xm nhp theo li m bo, to khng bo. Dung hp vi mng li ni cht hoc endosom tiu ha ri gii phng nucleocapsid (v d virus cm, toga v rabdo) 2. Virus khng c v ngoi. Xm nhp theo li m bo, to khng bo, dung hp vi endosom tiu ha, tin hnh ci v v gii phng acid nucleic (v d virus polio, adeno v reo).
Qu trnh nhn ln ca virus din ra theo 7 giai on

1. Hp ph - Gn th th c hiu ca mnh ln th th nm trn mng sinh cht ca t bo. V c tnh c hiu cao nn ch c virus nht nh mi gn ln c cc t bo nht nh. 2. Xm nhp - S hp ph c tng cng khi c mt ca ion Mg2+ hoc Ca2+
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- Virus khng c v ngoi : Mng t bo lm vo bao ly virus to khng bo tm thi. Tip khng bo dung hp cng vi mng li ni cht gii phng nucleocapsid. - Virus c v ngoi: V ngoi ca virus dung hp vi mng sinh cht ri y nucleocapsid vo trong m khng to khng bo. V ngoi virus ho vi mng sinh cht m khng chui vo t bo cht . - Mng t bo lm vo bao ly virus cng v ngoi, to khng bo. Sau mng khng bo (c ngun gc t mng t bo) dung hp vi v ngoi ca virus ri y nucleocapsid vo t bo cht.
Hnh 2.16. Virus khng c v ngoi v virus c mng bao 3. Ci v Enzym tiu ho ca t bo t lysosome tin hnh phn gii v protein cu virus gii phng acid nucleic hoc genome vo t bo cht. 4. Phin m - To thnh mRNA ca virus hoc phn t dng sao chp ca genome (RF- replicative form) 5. Tng hp cc thnh phn ca virus 5.1. Tng hp protein ca virus mRNA ca virus c phin m trn ribosome ca t bo to ra 2 loi protein - Protein cu trc l protein capsid, protein v ngoi v protein trong li. - Protein khng cu trc l enzym cn cho sao chp genome. Protein khng cu trc tm thy trong ht virus, tr mt s trng hp c bit v d enzym phim m ngc c trong virus HIV hoc virus vim gan B cha DNA polymerase, mt s virus RNA cha RNA polymerase 5.2.Tng hp acid nucleic ca virus - Genome ca virus con c sao chp t genome ca virus m. Trong trng hp genome l mch n th khun l mch b sung mi to thnh ca genome m. - Phn ln qu trnh sao chp c thc hin nh polymerase (replicase) do virus m ho. i vi mt s virus DNA th qu trnh tng hp c thc hin nh enzym ca t bo. 6. Lp rp - Genome v protein mi c to thnh lp rp vi nhau to nn ht virus mi. a s trng hp protein capsid lp rp to thnh cu trc rng gi l tin capsid (procapsid) sau do chuyn ng Brao, acid nucleic chui vo cu trc ny ri hn kn li.
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- Lp rp c th xy ra trong nhn t bo, trong t bo cht hoc ngay st mng sinh cht (i vi a s virus c v ngoi). Mng sinh cht bao ly nucleocapsid to v ngoi. 7. Gii phng Virus c th lm tan t bo chui t ra ngoi hoc i vi virus c v ngoi th chui ra t t theo li ny chi.
Hnh 2.17. Chu trnh sinh tan (lytic cycle) v tim tan (lysogenic cycle)
IX. CC PHNG THC NHN LN CA VIRUS

Qu trnh nhn ln ca virus trong t bo ht sc phc tp. Mi nhm virus c cch nhn ln ring. Sau y l vi nt n gin ho qu trnh nhn ln nu bt s khc nhau gia cc nhm.
1. Qu trnh nhn ln ca virus RNA chui dng
Hnh 2.18. S nhn ln ca virus RNA chui dng, khng to thnh cc mRNA genome.
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Sau khi xm nhp v ci v (bc 1), RNA genome c s dng trc tip tng hp cc polyprotein kch thc ln, cc protein cu trc v khng cu trc (bc 2 v 3). Protein khng cu trc (enzym) xc tc sao chp RNA thng qua s tng hp RNA chui m (i genome) (bc 5 v 6). Qu trnh sao chp to ra nhiu genome RNA mi phc v cho tng hp cc protein ca virus (bc 6 v 7) v cho lp rp to virus mi (bc 8). Mi tn m th hin s tng hp mc cao hn.
Hnh 2.19. S nhn ln ca virus RNA dng, km theo tng hp RNA di genome.
Sau khi xm nhp v ci v (bc 1), RNA genome c dng trc tip lm mRNA tng hp protein khng cu trc, trong c enzym RNA polymerase ph thuc RNA (RdRp) ( bc 2 v 3). RdRp xc tc sao chp RNA, tng hp chui RNA m (i genome) c kch thc (tng ng RNA genome) ( bc 4 v 5). T chui m lm khun tin hnh sao chp to ra nhiu chui RNA genome mi (bc 5 v 6). mRNA di genome (c kch thc nh hn genome) tin hnh dch m to protein cu trc ( bc 7 v 8). Genome mi to thnh tin hnh dch m to nhiu protein khng cu trc (bc 9 v 10) v sau lp rp vi protein cu trc to virus mi ( bc 11). Ch thch: Bc 7. Dch m ca mRNA di genome; Bc 8. Phn ct nh protease v x l sau dch m; Bc 10. Phn ct nh protease
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2. Qu trnh nhn ln ca virus RNA chui n, m
Hnh 2.20. S nhn ln ca virus RNA chui n, m

Sau khi xm nhp v ci v mt phn( bc 1), RNA tin hnh phin m nh RNA polymerase ph thuc RNA do virus mang theo to mRNA (bc 2) cho tng hp protein cu trc v khng cu trc (RdRp) ( bc 3). Enzym RdRp tin hnh sao chp RNA thng qua bc trung gian l to RNA chui dng (i genome) (bc 4 v 5). i genome (chui dng) c dng lm khun tng hp nhiu RNA genome (bc 5). Mt phn RNA genome c dng tin hnh phin m mnh, to ra mRNA cn cho tng hp protein (bc 7), mt phn dng lp rp vi protein to virus mi (bc 8).
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3. S nhn ln ca virus RNA chui kp
Hnh 2.21. S nhn ln ca virus RNA chui kp.

Sau khi xm nhp v ci v tng phn (bc 1), cc on RNA chui kp vn nm trong li tin hnh phin m nh enzym RdRp gn li tng hp mRNA (bc 2) ri t tng hp protein (bc 3). Cc protein ny lp rp quanh mRNA to ht di mc virus (bc 4). Sau mRNA trong cc ht ny tin hnh sao chp to RNA genome chui m tham gia to thnh RNA genome chui kp (bc 5). Cc ht di mc virus c to thnh li tip tc tham gia vo qu trnh tng hp m RNA (bc 6), tng hp protein (bc 7), sao chp (bc 8 v 9), lp rp vi protein cu trc to thnh virus mi (bc 10) Ch thch: Bc 1. Xm nhp v ci v Bc 4. Lp rp to ht di mc virus. Bc 10. Lp rp to virus mi.
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4. S nhn ln ca virus Retro
Hnh 2.22. S nhn ln ca virus Retro

Sau khi xm nhp v ci v tng phn (bc 1), RNA genome c phin m thnh cDNA (i genome), sau thnh DNA chui kp nh enzym phin m ngc (bc 2 v 3). Nh enzym integrase, DNA ca virus ci xen vo nhim sc th t bo ch (bc 4). C hai loi enzym trn u nm trong li ca virus. Nh RNA polymerase ca t bo, provirus phin m to RNA (bc 5) l tin cht ca mRNA (bc 6) tng hp protein (bc 7) v lp rp to virus mi (bc 8)
65
Cu hi n tp chng 2
1. iu no sau y ng khi ni v virus: a. L dng sng n gin nht b. L dng sng cha c cu to t bo c. Ch cu to t hai thnh phn c bn l protein v nucleic acid d. C a, b, c u ng. 2.Hnh thc sng ca virus l: -a. Sng k sinh khng bt buc -b. Sng cng sinh -c. Sng hoi sinh -d. Sng k sinh bt buc 3.Virus sinh sn theo kiu: -a. V tnh -b. Hu tnh -c. Tip hp -d. Da vo b my sinh nng lng v sinh tng hp ca t bo ch 4.Virus no sau y trong b gen ch cha RNA m khng c DNA ? -a. Virus gy bnh khm thuc l (TMV) -b. Virus HIV1, HIV2 -c. Virus SARS -d. C 3 dng virus trn 5.Con ng no c th ly truyn HIV - ng mu - ng tnh dc - Qua mang thai hay qua sa m nu m nhim HIV - C a, b, c u ng 6. Virus ch cha DNA m khng cha RNA -a. Virus gy bnh xon l c chua -b. Virus gy bnh hnh thoi c khoai ty -c. Virus H5N1 -d. C a, b, c u sai 7.Ln u tin, virus c pht hin trn: -a.Cy thuc l ; -b. Cy c chua -c. Cy khoai ty; -d. Cy u H Lan 8.Da vo hnh thi bn ngoi, virus c phn chia thnh cc dng no sau y: -a.Dng que, dng xon -b. Dng cu, dng khi a din, dng que -c. Dng xon, dng khi a din, dng que -d.Dng xon, dng khi a din, dng phi hp 9.Virus no sau y c dng khi ? -a. Th thc khun; -b. Virus gy bnh di
66
-c. Virus gy bnh bi lit; -d. Virus khm thuc l 10.Phage l virus ca vi khun c cu trc: -a.Dng xon ; -b. Dng khi -c. Dng que ; -d. Dng phi hp 11. Hy gii thch cc thut ng sau y: capsome, capsid, nucleocapsid, capsid c cu trc xon, khi a din, virus c cu trc phc tp, v ngoi. 12. Virus khc vi plasmit v khc vi t bo im no. 13. Virus c th tc ng ln t bo nh th no ? 14. Nu cc giai on nhn ln ca virus. 15. Ti sao virus ch c th nhn ln trong cc t bo nht nh ? 16. Trnh by cc phng thc xm nhp ca virus vo t bo ch. 17. Hy nu cc loi genome ca virus, Th no l RNA(+), RNA(-).Th no l genome chng lp. 18. Hy tm tt qu trnh nhn ln ca cc virus DNA n, DNA kp, RNA(+), RNA(-).
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT, H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc; NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty, 1997. Vi sinh vt hc, NXBGD, H Ni.
* Ti liu tham kho

1. Nguyn Thnh t, 1999. Vi sinh vt hc, NXBGD, H Ni. 2. http://vietsciences.net v http://vietsciences.free.fr 3. Lansing M. Prescott, john P. Harley, onal A. Klein, 2005. Microbiology. Mc Graw Hill.
* Gii thch thut ng

Prion: L nhng phn t protein v khng cha mt loi acid nucleic no. khi c lp protein ny, ngi ta c cu hnh cn bn ca n l mt dy protein gm 263 acid amin. Tim tan hay Tim sinh virus (ting Anh l lysogeny) l mt pha (phase) trong chu k sinh sn ca virus. Pha ny b sung vi pha tan (lytic phase), xy ra sau giai on xm nhim ca virus ng vt. Viroid: L nhng phn t RNA kn c 150 ribonucleotid, dng trn khng c v capsid. Pht hin nm 1971. Thc khun th (bacteriophage hay phage) c dng ch cc vt th xm nhim sinh vt nhn s (vi khun hoc vi khun c).
Chng 3: Sinh hc ca cc c th nhn s
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CHNG 3
SINH HC CA CC C TH NHN S
I. C KHUN (Archaea)
1. Khi nim
Archaea (ting Latinh, tc l "c i"; i khi c dch l vi khun c hay c khun) l mt phn nhm ln trong prokaryote (bn cnh vi khun). V v tr phn loi ca nhng sinh vt ny c nhiu im cha thng nht v chng c nhng c im ging vi vi khun nhng cng mang nhiu c im ca eukaryote.
[1]
[2]
[3] Hnh 3.1: Carl R. Woese [1], v tr phn loi Archaea [2] v h thng 3 lnh gii ca sinh vt [3].Vi khun (Bacteria), C khun (Archaea) v Sinh vt nhn thc (Eukarya). Cc Archaea m trc y gi l cc vi khun c (Archaeobacteria) l nhm c th nhn s c sm nht (khong 3,5-4 t nm trc y).
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Carl R. Woese v cng s (1977) sau khi xem xt trnh t 16S rRNA nhn thy rng cc sinh vt nhn nguyn thu (Prokaryote) cn c chia thnh hai nhm khc bit nhau hon ton l vi khun (Eubacteria hay Bacteria) v c khun (Archaeabacteria hay Archaea), v cng vi cc sinh vt nhn thc (Eukarya) lm thnh ba lnh gii (Domains) sinh vt. Cc nghin cu su hn v ph h v c im sinh l sinh ho cho thy rng c khun c tch ra t rt sm trong qu trnh tin ho, chng khng gn vi khun nhiu hn so vi sinh vt nhn tht, do vy tn gi Archaea c xut thay cho Archaeabacteria. Hin nay c hai tn gi Archaea v Archaeabacteria u c s dng trong cc ti liu vi sinh vt, tuy nhin thut ng Archaea chnh xc hn v r rng c khun khng phi vi khun m l mt nhm vi sinh vt ring bit.
2. c im ca c khun
C khun l mt nhm VSV c bit, c nhiu c im rt khc bit (Bng 3.1). Bng 3.1. Nhng c im khc bit ca c khun so vi vi khun v sinh vt nhn thc Vi khun Sinh vt nhn c im C khun(Archaea) (Bacteria) thc (Eukarya) Pseudo-peptidoglycan, cellulose, Thnh t bo Peptidoglycan protein, polysaccharid, carbonat, silicat, glycoprotein chitin Mng t bo Este-lipid Ethe-lipid Este-lipid Ribosome 70 S 70 S 80 S Phn ng ca ribosome khng Mn cm Mn cm vi c t bch hu Cng nh t bo vi khun, t bo c khun (ngoi tr chi Thermoplasma) c thnh t bo bn ngoi gi chc nng bo v. Tuy nhin, khng nh vi khun, thnh t bo ca c khun khng cha peptidoglycan v v th khng b ph hu di tc dng ca lysozym. C khun c rt nhiu dng cu trc thnh t bo khc nhau. Mt s c khun (nh cc loi sinh methane) c thnh t bo cu to bi mt loi polysaccharid rt ging vi peptidoglycan c gi l pseudo-peptidoglycan (pseudomurein). Chui pseudo-peptidoglycan gm cc n nguyn N-acetyl-glucosamin v N-acetyl-alosamin-uronic acid (thay cho N-acetyl-muramic acid trong peptidoglycan). Ngoi ra, y cu ni glycosid 1 3 thay th cho cu ni glycosid 1 4 peptidoglycan. Mt s c khun khc li hon ton khng c c peptidoglycan v pseudo-peptidoglycan trong thnh t bo m thay vo l hn hp gm polysaccharid, glycoprotein hoc protein. V d nh cc loi Methanosarcina (c khun sinh methane) c thnh t bo l mt lp polysaccharid dy cu to t glucose, glucuronic acid, galactosamin v Acetate. Cc loi c khun a mn cc oan (extreme halophiles) nh l Halococcus c thnh t bo tng t nh Methanosarcina nhng cha nhiu hp cht c nhm sulfat ging nh chondroitin sulfat t chc lin kt ca ng vt. Dng cu trc thnh t bo ph bin nht c khun l lp paracrystallin b mt (S-layer) gm protein hay glycoprotein. Cu trc ny c tm thy cc i din thuc tt c cc nhm c khun, t a mn cc oan (extremely halophilic), a nhit cc oan (extremely thermophilic) v c cc loi sinh methane. c bit cc chi Methanospirillum v Methanothrix (c khun sinh
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methane) c cu trc thnh t bo v cng phc tp. Cc loi thuc hai chi ny mc thnh chui di gm nhiu t bo, gia mi cp t bo c mt lp m dy v ton b cu trc chui li c bc kn trong mt lp paracrystallin b mt. Thnh phn v cu trc lipid ca mng t bo l mt trong nhng c im ni bt phn bit c khun v hai nhm cn li. Trong khi vi khun v sinh vt nhn tht cu ni acid bo glycerol trong lipid mng t bo l lin kt este (ester) th c khun li l lin kt ete (ether) (Hnh 3.2). Acid bo trong este-lipid thng l cc phn t ngn, mch thng. Tri li, acid bo trong ete-lipid l cc phn t mch di, phn nhnh, thuc c hai dng phytanyl (C20 cacbuahydro tng hp t isopren) v biphytanyl (C40). Do ch c c khun v khng b bin i di nhit cao nn isopren-lipid c ly lm cht ch th ca c khun ho thch. Enzym polymeraza thc hin qu trnh sao m trn khun DNA (DNA-dependent RNA polymerase) ba lnh gii sinh vt cng c nhiu im khc nhau. Vi khun ch c mt loi ARN-polymeraza c cu trc khng gian n gin, gm bn chui polypeptid 2 , 1 , 1 v mt nhn t khng c nh. C khun c nhiu loi ARN-polymeraza, cu trc mi loi li phc tp hn nhiu so vi ARN-polymeraza vi khun. ARN-polymeraza ca c khun sinh methane v cc loi a mn (halophilic) gm tm chui polypeptid (5 chui di v 3 chui ngn). ARN-polymeraza c khun a nhit cao (hyper-thermophilic) li phc tp hn, gm t nht 10 chui peptid. Polymeraza thc hin qu trnh tng hp ARN thng tin (mARN) sinh vt nhn tht gm 10-12 chui polypeptid c kch thc tng t nh ARN-polymeraza ca c khun a nhit cao. Ngoi ra, sinh vt nhn tht cn c hai loi ARN-polymeraza khc na c hiu cho qu trnh tng hp ARN ca ribosom (rARN) v ARN vn chuyn (tARN). Nh vy cht khng sinh rifampicin c tc dng c ch n v ca polymeraza ch c hiu qu i vi vi khun v c khun v sinh vt nhn tht khng c loi polymeraza ny. Vi nhng im khc bit trong trnh t 16S rARN cng nh cu trc ARNpolymeraza, hin nhin b my sinh tng hp protein ca ba lnh gii sinh vt cng s khng ng nht. Tuy c kch thc ca ribosom ging vi vi khun (70S) nhng c khun li c nhiu bc trong qu trnh sinh tng hp protein rt ging vi sinh vt nhn tht (80S ribosom). Nhiu cht khng sinh c ch qu trnh sinh tng hp protein vi khun li khng c hiu lc i vi c khun v sinh vt nhn tht (Bng 3.2). Ngoi ra, tng t nh sinh vt nhn tht, nhn t ko di EF-2 trong ribosom c khun c phn ng vi c t bch hu, mt loi c t v hi i vi vi khun. Tuy nhin nhn t EF-2 c khun mang tnh c hiu cao, nhn t ny hon ton khng hot ng trong mi trng ribosom ca vi khun hoc sinh vt nhn tht. Cc th nghim lai ribosom in vitro cho thy ribosom ghp gia n v ln (50S) ca c khun v n v nh (40S) ca sinh vt nhn tht vn thc hin chc nng gii m mt cch bnh thng, trong khi vic ghp tng t gia vi khun v sinh vt nhn tht li hon ton khng tng thch. Nh vy cu trc b my sinh tng hp protein ca c khun c nhiu im tng ng vi sinh vt nhn tht hn l vi vi khun. Ging nh vi khun, c khun c mt nhim sc th dng vng, tuy nhin genom ca c khun thng nh hn nhiu so vi genom ca vi khun. Chng hn DNA ca Escherichia
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coli l 2,5 x 109 Da, trong khi DNA ca Thermoplasma acidophilum l 0,8 x 109 Da, hay ca Methanobacterium l 1,1 x 109 Da. Ngoi ra thnh phn GC (mol%) ca DNA c khun dao ng trong phm vi rt ln, t 21 n 68 %, chng t tnh a dng ca c khun. So snh trnh t y ca genom c khun Methanococcus jannaschi vi genom ca vi khun v sinh vt nhn tht cho thy 56% trong 1738 gen khng tng ng.
Hnh 3.2: Lipid trong mng t bo ca c khun (ete-lipid) khc vi ca vi khun v sinh vt nhn thc (este-lipid) Bng 3.2. Tnh mn cm ca i din ba lnh gii sinh vt i vi cc cht c ch qu trnh sinh tng hp protein
C khun Cht khng sinh Cycloheximid Virginiamycin, pulvomycin Neomycin, puromycin Rifamycin Erythromycin, streptomycin, chloramfenicol Tc dng c ch c ch bc khi u c ch bc ko di Dng tng hp sm c ch enzym RNA polymerase Tng tn s mc li v mt s hiu ng khc Vi khun Sinh vt nhn thc
Methanobacterium
Sulfolobus
Escherichia coli
Saccharomyces cerevisae
~ + + ~ ~
~ ~ + ~ ~
~ + + + +
+ ~ + ~ ~
3. Mi trng sng ca c khun v gi thuyt v hnh thnh s sng trn tri t

C khun c bit n nh nhng vi sinh vt thch nghi vi cc mi trng c iu kin cc oan (extreme) nh nhit cao (thermophilic), ni lnh gi (psychrophilic), nng mui cao (halophilic) hay acid cao (acidophilic) v.v. cng l mt l do gii thch ti sao c khun li kh c phn lp v nui cy trong iu kin phng th nghim. Trong gii sinh vt, c khun c cc i din c tr cc iu kin nhit cao hn c (Bng 3.3, Hnh 3.3). Bng 3.3. Nhit pht trin cao nht ca cc i din sinh vt trn tri t
Cc i din sinh vt trn tri t Nhit pht trin cao nht oC
C Cn trng ng vt n bo
38 50 50
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To 56 Nm 60 Vi khun thng 90 C khun 113 Vi cc c im sinh l nh tnh a nhit, sng k kh, s dng cc cht hu c v v c l ngun nng lng, cc loi c khun a nhit cao c l ph hp vi dng sng nguyn thu m phng theo iu kin ca tri t trong thi k u. Trong thc t, cht ch th mch isoprene-lipid thnh phn mng t bo ca c khun c tm thy trong cc lp trm tch c tui l 3,8 t nm. Cc nghin cu da trn trnh t 16S rRNA cho thy c khun, c bit l nhm c khun a nhit cao, tin ho chm hn ng k so vi vi khun v sinh vt nhn thc. Tuy nhin tc tin ho chm ca c khun so vi hai lnh gii cn li c th do mi trng sng khc nghit ca chng to ra. Cho n nay cu hi v ngun gc s sng v vai tr ca c khun trong vn cn ang tip tc c tranh lun.
Hnh 3.3. Mt trong nhng ni u tin c khun c tm thy: sui nc nng trong cng vin Quc gia Yellowstone (M).
Hnh 3. 4. Cc i din ca hai nhm c khun Crenarchaeota v Euryarchaeota.
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Hnh 3.5: Hnh thi mt s i din ca hai nhm c khun Euryarchaeota v Crenarchaeota
4. Ph h c khun da trn trnh t 16S rRNA

Da trn so snh trnh t 16S rRNA cc i din c khun phn lp c chia thnh hai nhm chnh l Euryarchaeota v Crenarchaeota (Hnh 3.4, H.3.5). Euryarchaeota l nhm c khun c bit r nht, bao gm nhiu loi sinh methane, c khun a mn, kh sulfat (Archaeoglobales), Thermoplasmalates v Thermococcales. Nhm Crenarchaeota gm ba lp Desulfococcales, Sulfolobales v Thermoproteales. Sau ny nhm c khun Korarchaeota c xut thm (Hnh 3.6), tuy nhin ch da trn cc trnh t 16S rDNA c c t cc mu DNA tch trc tip t mi trng ch cha c i din no c phn lp v nui cy trong phng th nghim. Nm 2002, nhm nghin cu ca gio s Stetter, mt trong nhng nh nghin cu c khun hng u th gii, cng b s hin din ca nhm c khun th 4, Nanoarchaeota, gm nhng c khun c kch thc rt nh vi mt i din duy nht c tm thy l Nanoarchaeum equitans (Hnh 3.7). Loi c khun ny c t bo hnh cu, ng knh 400 nm, sng bm trn b mt t bo ca mt loi c khun mi Ignicoccus sp., phn lp t mu nc nng su 106 m di y bin. y l mt loi a nhit cc oan, pht trin nhit ti u 75-980C. Nhiu trnh t 16S rARDN trc tip c c t mi trng c nhit cao cng khng nh s tn ti v khc bit ca nhm Nanoarchaeota so vi cc nhm c khun cn li.
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Hnh 3.6. Mi lin quan ph h ca ba nhm c khun Euryarchaeota v Crenarchaeota v Korarchaeota
Hnh 3.7. Nanoarchaeum equitans (cu khun nh) trn b mt Ignicoccus sp. (cu khun ln)
5. Cc hnh thc dinh dng c khun

C khun c nhiu hnh thc dinh dng: ho dng hu c (chemoorganotrophy), ho dng v c (chemolithotrophy), t dng (autotrophy), hay quang hp (phototrophy). Ho dng hu c l hnh thc dinh dng ca nhiu loi c khun, tuy nhin cc chu trnh phn gii cht hu c thng c mt s im khc bit so vi vi khun. C khun a mn (halophiles) v a nhit cc oan (extreme thermophiles) phn gii glucose theo mt dng ci bin ca con ng Entner-Doudoroff (E-D). Nhiu loi c khun li c kh nng sn sinh ra glucose t cc cht ban u khng phi l hydratcarbo (gluconeogenesis) thng qua cc bc o ngc ca qu trnh glycolysis (con ng Embden-Meyerhof). Oxygen ho Acetate thnh CO2 c thc hin qua chu trnh TCA (i khi vi mt s thay i trong cc bc phn ng), hoc qua con ng acetyl-CoA (Ljungdahl-Wood). Cc thnh phn ca chui vn chuyn in t nh vi khun u c tm thy c khun, trong cytochrom a, b v c c cc loi a mn cc i, cytochrom a c mt s loi a nhit cao. M phng da trn chui chuyn in t phn ln c khun cho thy chng thu np in t t cht cho vo chui nc thang NADH, oxygen ho cht nhn in t cui cng l O2, S0 hay mt s cht khc, ng thi to ra lc y proton (proton motiv force) tng hp ATP nh b my ATPase c tr trong mng t bo. Ho dng v c kh ph bin c khun, trong hydrogen thng c s dng lm cht cho in t. T dng c bit ph bin c khun v din ra di nhiu hnh thc khc nhau. c khun sinh methane v c khun ho dng v c a nhit cao CO2 c chuyn ho thnh cc hp cht hu c qua con ng acetyl-CoA, trong mt s loi c ci bin cc bc phn ng khc nhau. Mt s loi c khun khc (nh Thermoproteus) c nh CO2 theo chu trnh citric acid o ngc, tng t nh vi khun lam lu hunh. Mc d cc loi c
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khun a nhit cc oan u thc hin hnh thc dinh dng hu c nhng nhiu loi vn c kh nng c nh CO2 v thc hin qu trnh ny theo chu trnh Calvin, tng t nh vi khun v sinh vt nhn tht. Kh nng quang hp c mt s loi c khun a mn cc oan, tuy nhin khc vi vi khun, qu trnh ny c thc hin hon ton khng c s tham gia ca chlorophill hay bacteriochlorophill m nh mt loi protein mng t bo l bacteriorhodopsin kt gn vi phn t tng t nh carotenoid c kh nng hp ph nh sng, xc tc cho qu trnh chuyn proton qua mng nguyn sinh cht v s dng tng hp ATP. Tuy nhin, bng hnh thc quang hp ny c khun a mn cc oan ch c th sinh trng vi tc thp trong iu kin k kh, khi mi trng thiu cht dinh dng hu c.
6. Mt s nhm c khun i din
Xt v cc c im sinh l, c khun c th phn thnh bn nhm chnh l:

a. Sinh methane (methanogens) b. C khun a nhit cao (hyper-therrmophiles) c. C khun a mn (halophiles) v d. C khun a acid (acidophiles) thuc lp Thermoplasmatales vi nhiu i din c phn lp v nghin cu trong phng th nghim.
Hnh 3. 8. i din cc nhm c khun chnh 6.1. C khun sinh methane (methanogens) Trong c khun, cc loi sinh methane lm thnh mt nhm ln v a dng vi cc
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c im chung l : (1) To kh methane nh sn phm cui cng ca chu trnh trao i nng lng v (2) Sng k kh bt buc. C khun sinh methane thu nng lng cho qu trnh sinh trng t vic chuyn ho mt s cht thnh kh methane. Ngun c cht ch yu ca cc vi sinh vt ny l hydro, format v Acetate. Ngoi ra, mt s hp cht C1 nh metanol, trimethylamin, dimethylsulfid v ru nh isopropanol, isobutanol, cyclopentanol, etanol cng c s dng lm c cht (Bng 3.4). Qu trnh sinh methane c khun c th coi nh l qu trnh h hp k kh, trong cht nhn in t l CO2 hoc nhm methyl trong cc hp cht C1 v Acetate. Tuy nhin, nh ta thy trong bng 4, nng lng c gii phng ra trong cc phn ng to methane u rt nh. so snh ta c th ly nng lng gii phng t phn ng oxygen ho glucose bng oxygen C6H12O6 + 6 O2 6 CO2 + 6 H2O (G0 = 2870 kJ/mol). L nhng sinh vt duy nht c kh nng to ra kh methane, c khun sinh methane c nhng enzym v coenzym thit yu cho qu trnh tng hp methane v ng vai tr ch th cho nhm, v d nh coenzym F420 v coenzym M. S hin din ca coenzym F420 khin cho cc t bo ca c khun sinh methane c tnh t pht sng di nh n hunh quang (bc sng 350 420 nm). Mc d hin tng t pht sng ny c th mnh, yu, hay i khi mt hn, tu thuc vo cc pha sinh trng ca t bo, nhng vn l mt c im n gin v tin li nhn bit c khun sinh methane di knh hin vi. Bn cnh , trnh t acid amin trong cc chui peptid ca coenzym M cng c dng phn loi c khun sinh methane. Nhng nghin cu trong lnh vc ny cho thy s tng ng gia cy phn loi da trn trnh t 16S rARN v cy phn loi da trn trnh t acid amin ca cc n v v trong coenzym M. Bng 3.4. Phn ng to methane trn cc c cht khc nhau v nng lng c gii phng t Phn ng sinh methane Nng lng c gii phng G0 (kJ/ methane) -135,6 4H2 + CO2 CH4 + 2H2O -130,1 4 Format CH4 + 3CO2 + 2H2O - 36,5 4 Isopropanol + CO2 CH4 + 4 Aceton + 2H2O -116,3 2 Etanol + CO2 CH4 + 2 Acetate -112,5 Metanol + H2 CH4 + H2O -104,9 4 Metanol 3CH4 + CO2 + 2H2O + - 75,0 4 Methylamin + 2H2O 3CH4 + CO2 + 4NH4 + - 73,2 2 Dimethylamin + 2H2O 3CH4 + CO2 + 2 NH4 + - 74,3 4 Trimethylamin + 6H2O 9 CH4 + 3CO2 + 4 NH4 - 73,8 2 Dimethylsulfid + 2H2O 3CH4 + CO2 + H2S - 31,0 Acetate CH4 + CO2 Nhng ni thng thng c th tm thy c khun sinh methane l cc b ln men hu c k kh, cc lp trm tch thiu oxygen, t ngp ng v h ng rut ca ng vt. Khi dng chng n c khun sinh methane rt nhy cm vi oxygen, tuy vy trong t nhin chng c th tn ti mi trng hiu kh nh c bao bc v bo v bi cc vi sinh
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vt hiu kh v k kh khc. Trong mi trng k kh, c khun sinh methane phi cnh tranh v c cht, c bit l hydro v Acetate, vi cc nhm vi sinh vt s dng cht nhn in t c hiu in th kh dng tnh hn so vi CO2 nh l nitrat, sulfat v oxygent st III. Nh vy c khun sinh methane s chim lnh cc mi trng ni khng c nhiu cc loi cht nhn in t tim nng ny. Do khng c kh nng s dng rng ri cc loi c cht khc nhau, trong t nhin c khun sinh methane thng phi ph thuc vo cc loi vi khun ln men v chng chuyn ho a dng cht hu c thnh cc acid hu c, hydro, format v Acetate, trong hydro, format v Acetate l ngun thc n trc tip cho c khun sinh methane, cn cc acid hu c sn phm ca qu trnh ln men nh propyonat, butyrate th cn phi c mt nhm vi khun khc chuyn ho thnh c cht thch hp ri mi n lt c khun chuyn thnh kh methane. C hai hnh thc cng sinh: bt buc v khng bt buc. Trong hnh thc cng sinh gia c khun sinh methane v vi khun ln men, ch c c khun ph thuc vo mi lin h ny do nhu cu v thc n cn cc hot ng trao i cht ca chng hon ton khng c nh hng g ti cc vi khun ln men, v th hnh thc ny c gi l cng sinh khng bt buc. Cng sinh bt buc din ra gia c khun sinh methane v mt nhm vi khun cng sinh bt buc, trong c i bn cng cn n nhau, v d nh hin tng cng sinh gia Methanobrevibacter v Synthrophobacter (Hnh 3.9). Nhm vi khun cng sinh trong mi lin kt ny oxygen ho acid hu c nh propionate, cc acid c mch carbon di hn, cc hp cht thm v chuyn in t sang proton hay CO2, to thnh hydro hay format tng ng. Nhm vi khun cng sinh ny ch c th thc hin c trao i cht khi nng hydro v format trong mi trng xung quanh c gi mc rt thp, v nhim v do c khun sinh methane m nhim. Cc loi sinh methane thng c mt trong mi lin kt cng sinh bt buc l Methanoplanus endosymbiosus, cc loi Methanobrevibacter, v Methanobacterium formicicum.
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mt s mi trng c bit nh cc sui nc nng hay cc tng nham thch ni la c khun sinh methane thng c mt vi s lng ln. Trong trng hp ny chng sng t do, khng ph thuc vo cc vi sinh vt khc v ngun c cht ch yu c s dng y l hydro, sn phm c c t con ng l ho ch khng qua con ng sinh hc. Hin nay tng s c khun sinh methane c bit n l 50 loi thuc 19 chi, su h v ba lp (Bng 3.5 v 3.6). S phn loi ny da trn trnh Hnh 3.9 : Cng sinh gia Methanobrevibacter (t bo trc khun) v Synthrophobacter (t bo hnh oval). t 16S rRNA hin c, tuy nhin chng ta c th hnh dung c rng theo thi gian khi nhng loi mi c bit thm th c th bng phn loi ny s cn phi thay i. V hnh thi, c khun sinh methane rt a dng, trong mt s loi c hnh dng c trng d nhn bit di knh hin vi nh Methanosarcina, Methanospirillum hay Methanosaeta (Hnh 3.8). Ngoi ra, mt trong nhng c im quan trng dng phn loi nhm c khun ny l ngun c cht c th s dng sinh methane. H Methanobacteriaceae c thnh t bo cu to t pseudomurein, v th bt mu Gram (+). H Methanobacteriaceae gm c ba chi l Methanobacterium, Methanobrevibacter v Methanosphaera. Cc loi thuc chi Methanobacterium c t bo hnh que hoc hnh si, i khi to nhm gm nhiu t bo. Tt c cc loi thuc chi ny u c kh nng sinh methane t H2 + CO2. Bng 3.5. Cc nhm phn loi chnh ca c khun sinh methane Lp H Chi Methanobacteriales Methanobacteriaceae (T) Methanobacterium (T) -nt-nt-ntMethanococcales Methanomicrobiales -nt-nt-nt-ntMethanothermaceae Methanococcaeae (T) Methanosarcinaceae -nt-ntMethanobrevibacter Methanosphaera Methanothermus (T) Methanococcus (T) Halomethanococcus Methanococcoides Methanohalobium
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-nt-nt-nt-nt-nt-nt-nt-nt-nt-nt-nt-
-nt-nt-nt-ntMethanomicrobiacaea (T) -nt-nt-nt-nt-ntMethanocorpusculaceae
Methanohalophilus Methanolobus Methanosarcina (T) Methanosaeta (Methanothrix) Methanoculleus Methanogenium Methanolacinia Methanomicrobium (T) Methanoplanus Methanospirillum Methanocorpusculum (T)
T = h/chi chun; -nt- nh trn Bng 3.6. Mt s c im hnh thi v sinh l ca cc h C khun sinh methane chun H C khun c im C hai dng t bo: - Trc khun, di t bo khc nhau; s dng H2 + CO2, i khi format v ru lm ngun c cht sinh methane. - Cu khun, ch s dng H2 + metanol; a s l Gram(+); thnh t bo cha pseudomurein; khng chuyn ng; thnh phn GC = 23-61 mol% Trc khun, c cht sinh metan l H2+CO2; Gram(+); thnh t bo cha pseudomurein; khng chuyn ng; a nhit cc i (extreme thermophiles); thnh phn GC = 33-34 mol% Cu khun hnh dng khng u; c cht to methane l H2+CO2 v format; Gram(-); chuyn ng; thnh phn GC = 3961 mol% T bo hnh que, xon, hnh a hoc cu khng u; c cht sinh methane l H2+CO2, format, i khi l ru; Gram(-); c loi chuyn ng, c loi khng; thnh phn GC = 39-61 mol% Cu khun nh, khng u; c cht sinh methane l H2+CO2, format, i khi l ru; Gram(-); c loi chuyn ng, c loi khng; thnh phn GC = 48-52 mol% Cu khun khng u, trc khun c v bc bn ngoi; c cht sinh methane l H2+CO2, Acetate, cc hp cht c nhm methyl, khng s dng format; Gram (+) hoc (-); thng khng chuyn ng; thnh phn GC = 36-52 mol%
Methanobacteriaceae
Methanothermaceae
Methanococcaceae
Methanomicrobiaceae
Methanocorpusculaceae
Methanosarcinaceae
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C khun sinh methane c ng dng rng ri trong nhiu lnh vc khc nhau, in hnh nht l trong cc cng ngh x l cht thi giu hu c, chuyn ho cht hu c thnh methane v CO2. Do kh nng s dng c cht hn hp nn c khun sinh methane t c a vo cc qui trnh x l dng chng n m thng dng hn hp vi cc loi d dng c kh nng chuyn ho cht hu c trong cht thi thnh ngun c cht thch hp cho chng. Cc loi vi sinh vt c s dng ng thi vi c khun sinh methane trong cc qui trnh x l cht thi hu c thng l cc loi c kh nng ln men ng, protein nh Lactobacillus, Eubacterium, Clostridium, Klebsiella, hay Leuconostoc. Ngoi ra, c khun sinh methane cn c s dng phn hu cc hp bn vng lm nhim mi trng nh cc hp cht thm v cao phn t mch thng cha halogen. Cng vi vi khun kh sulfate, c khun sinh methane cn l tc nhn gy n mn kim loi iu kin khng c oxygen.
Methanothermus
Methanococcus
Methanosarcina
Methanospirillum
Methanomicrobium
Methanobacterium thermoautophicum
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Methanogenium
Methanoculleus
Methanosaeta
Hnh 3.8. Hnh thi t bo ca mt s loi c khun sinh methane 6.2. C khun a mn Halobacteriales y l mt nhm c khun a dng, thuc lp Halobacteriales, sng cc mi trng c nng mui cao nh l cc h nc mn trong t nhin, cc cnh ng mui hay trong thc phm mui nh tht mui, c mui lm nc mm. Cu to ho hc ca thnh t bo Halobacterium l glycoprotein c bit giu cc acid amin mang in tch dng nh aspartate v glutamate. Nhm carboxy ca cc acid amin ny lin kt vi ion Na+ (hng ra pha ngoi), v v th khi thiu Na+ thnh t bo s lp tc b ph v. Bng 3.7. Nng cc ion bn trong t bo Halobacterium salinarum Ion Nng ngoi mi trng (M) Nng trong t bo (M) Na+ K
+ 2+ -
3,3 0,05 0,13 3,3
0,8 5,3 0,12 3,3
Mg
Cl
C khun a mn c hnh thi rt a dng (Hnh 3.9), t hnh que, hnh cu cho n cc hnh dng v cng c bit. V ph h, da trn trnh t 16S rRNA c khun a mn phn nhnh rt gn vi c khun sinh methane.
Halobacterium
Haloferax
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Natronobacterium Natronococcus Haloarcula Hnh 3.9. Hnh thi mt vi i din c khun a mn Cc loi c khun a mn c bit n hin nay c xp vo 14 chi vi 35 loi c m t chnh thc, trong 6 chi ng thi l c khun a kim cc oan (extreme alkaliphiles). Mt c im thch nghi vi nng mui cao khc c khun a mn cc oan l phn ln cc protein trong nguyn sinh cht u phn cc, c tnh acid cao v hot tnh ph thuc vo ion K+. Trong mi trng c nng ion rt cao nh nguyn sinh cht c khun a mn cc oan th ch c protein phn cc mi tn ti c trong dung dch, cc protein khng phn cc iu kin ny s b kt dnh v mt hot tnh. 6.3. C khun a nhit cao (hyperthermophiles)
Sulfolobus
Acidianus
Thermoproteus
Thermofilum
Desulfurococcus
Pyrobaculum
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Pyrodictium occultum
Pyrococcus
Pyrolobus
Thermoproteus tenax
Staphylothermus marinus
Thermococcus
Hyperthermus Metallosphaera Archaeoglobus Hnh 3.10. Mt s i din c khun a nhit cao C khun a nhit cao l nhng sinh vt c kh nng sng iu kin nhit cao hn c trong gii sinh vt, chng u c nhit sinh trng ti u trn 80 C, nhiu loi c th sinh trng nhit nc si hoc cao hn na. Tu thuc vo c im a cht ca vng m mi trng solfatar mang tnh kim hoc acid nh (pH 5-8). Trong mt s trng hp pH trong mi trng xung rt thp do acid sulfurich c to ra t vic oxygen ho H 2S v S0 bi mt s loi vi khun v c khun. Ngoi tr mt vi trng hp, a s c khun a nhit cao sinh trng k kh bt buc v thc hin dinh dng theo phng thc ho dng hu c hoc ho dng v c. Hu ht cc loi u c nhu cu v lu hunh, hoc lm cht nhn in t trong h hp k kh, hoc lm cht cho in t trong h hp hiu kh. Cc loi k kh c th s dng cc hp cht hu c v v c (nh H2) kh lu hunh S0 thnh H2S. Ngc li, cc loi hiu kh oxygen ho H2S, S0 thnh acid sulfuric. Ngoi ra, mt s loi c kh nng thc hin h hp k kh s dng cc cht nhn in t khc ngoi S0 nh l nitrat (Pyrolobus, Pyrobaculum), sulfat (Archaeoglobus) hay sinh methane (Methanopyrus). V hnh thi, c khun a nhit cao tng i a dng, nhiu loi c hnh dng t bo c bit (Hnh 3.10). c loi sng nhng vng ni la di y i dng vi nhiu i din sinh trng trn 100 C nh p lc ca nc bin (Bng 3.9).
Aeropyrum
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6.4. C khun a nhit v acid (Thermoplasmatales) y l nhng vi sinh vt a nhit v pH thp (thermoacidophilic), h hp k kh khng bt buc, thng s dng cc hp cht hu c lm c cht nhng cng c loi c kh nng sinh trng t dng. Lp Thermoplasmatales gm ba h, mi h mi ch c mt chi i din, l h Thermoplasmaceae vi chi Thermoplasma, h Picrophilaceae vi chi Picrophilus v h Ferroplasmaceae vi chi Ferroplasma. Tt c cc i din ca Thermoplasmatales u bn vng vi cc cht c ch thnh t bo nh ampicillin, vancomycin v streptomycin. Chi Thermoplasma hin c hai loi, T. acidophillum phn lp t bi thi khai thc than v T. volcanium phn lp t sui nc nng (Hnh 3.11) Chi Picrophilus c hai loi i din l P. oshimae v P. torridus, s khc bit gia hai loi rt nh, ch da trn 3% khc bit v trnh t 16S rRNA v phn tch DNA bng enzym gii hn. C hai loi thuc chi ny u c phn lp t mi trng solfatar (nhit cao, giu hp cht lu hunh) Nht bn. Hai loi u b c ch nng NaCl 0,2M, chng t chng c ngun gc t t lin. Chi Ferroplasma cng gm hai loi, F. acidiphilum v F. acidarmanus, tuy nhin mi ch c loi F. acidiphilum c m t v a vo danh sch phn loi chnh thc. Hai loi ny khc nhau kh nng sinh trng trn chit xut nm men v pH ti u, cn trnh t 16S rRNA ca chng hon ton ging nhau. Ferroplasma c phn lp t cc b tch vng t qung arsenopyrit/pyrite Kazacstan.
Thermoplasma acidophilum
Thermoplasma volcanium
Hnh 3.11. i din chi Thermoplasma
Kt lun:
C khun l mt nhm vi sinh vt a dng, khng nhng v hnh thi v sinh l, m cn v mi trng sng, trong nhiu ni c iu kin v cng khc nghit m chi c khun mi c kh nng tn ti c. Tuy nhin nhng loi c khun c chng ta bit n v trnh by y mi ch chim mt phn rt nh ca ton b lnh gii. Hin nay vi cc k thut sinh hc phn t tin tin ngi ta c th tin hnh nghin cu v thu thp mt lng thng tin ng k v c khun m khng cn thng qua con ng phn lp v nui cy c truyn. Cc trnh t 16S rDNA c c trc tip t mi trng cho thy tnh a dng cao v s c mt ca c khun nhiu mi trng sng khc nhau, chng t c khun c vai tr rt quan trng trong h sinh thi trn tri t m hin nay con ngi cn cha hiu r c.
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II. VI KHUN (BACTERIA)

1. Khi nim chung:
Vi khun (ting Anh v ting La Tinh l bacterium, s nhiu bacteria) i khi cn c gi l vi trng. Vi khun l mt nhm sinh vt n bo, c kch thc nh (kch thc hin vi) v thng c cu trc t bo n gin khng c nhn, b khung t bo (cytoskeleton) v cha c cc bo quan nh ty th v lc lp. Vi khun l nhm hin din ng o nht trong sinh gii. Chng hin din khp ni trong t, nc v dng cng sinh vi cc sinh vt khc. Nhiu tc nhn gy bnh (pathogen) l vi khun. Chng thng c vch t bo vi thnh phn cu to l (peptidoglycan). Nhiu vi khun di chuyn bng tin mao (flagellum) c cu trc khc vi tin mao ca cc nhm khc. 2. Lch s nghin cu v phn loi vi khun Nm 1595 Zacharias Janssen ln u tin lp ghp knh hin vi nhng cha quan c vi khun. Vi khun u tin c quan st bi Antony van Leeuwenhoek nm 1683 bng knh hin vi mt trng do ng t thit k. Tn "vi khun" c ngh sau kh lu bi Christian Gottfried Ehrenberg vo nm 1828, xut pht t ch trong ting Hy Lp c ngha l "ci que nh". Louis Pasteur (1822-1895) v Robert Koch (1843-1910) miu t vai tr ca vi khun l cc th mang v gy ra bnh hay tc nhn gy bnh. Ban u vi khun hay vi trng (microbe) c coi l cc loi nm c kch thc hin vi (gi l Schizomycetes), ngoi tr cc loi vi khun lam (Cyanobacteria) quang hp, c coi l mt nhm to (gi l cyanophyta hay to lam). Phi n khi c nhng nghin cu v cu trc t bo th vi khun mi c nhn nhn l mt nhm ring khc vi cc sinh vt khc. Vo nm 1956 Hebert Copeland phn chng vo mt gii (kingdom) ring l Mychota, sau c i tn thnh Sinh vt khi sinh (Monera), Sinh vt nhn s (Prokaryota), hay Vi khun (Bacteria). Trong thp nin 1960, khi nim ny c xem xt li v vi khun (by gi gm c cyanbacteria) c xem nh l mt trong hai nhm chnh ca sinh gii, cng vi sinh vt nhn thc. Sinh vt nhn thc c a s cho l tin ha t vi khun, v sau cho rng t mt nhm vi khun hp li. 3. Hnh dng v kch thc ca vi khun Vi khun c nhiu hnh dng khc nhau. a s c hnh que, hnh cu, hay hnh xon; nhng vi khun c hnh dng nh vy c gi theo th t l trc khun (bacillus), cu khun (coccus), v xon khun (spirillum). Mt nhm khc na l phy khun (vibrio) c hnh du phy (hnh III.1.13). Ty theo s lin kt ca cc t bo sau khi phn ct m cu khun c th chia thnh cc loi song cu, lin cu, t cu.ng knh ca cu khun thng vo khong 0,5-1 m. Trc khun thng c kch thc thay i trong khong chiu rng 0,5-1,0 v chiu di 1,0-4,0 m, mc d c loi c ng knh n 0,3mm (Thiomargarista).
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Hnh 3.12. Cc nhm hnh thi chnh ca vi khun A.Cu khun; B.Trc khun, C.Xon khun; D. Chui cu khun; E. T cu khun; Khun mao, G. Anabaena; H. Oscillatoria; I. Gleocapsa.
4. Cu trc t bo vi khun
4.1. V nhy v dch nhy Nhiu vi khun c bao bc bn ngoi bng mt lp v nhy c bn cht ha hc l polysaccharid ngoi tr vi khun than l polypeptid. Khi lm kh, ngi ta xc nh c 90 98% trng lng ca mng nhy l nc.
Hnh 3.13. S cu trc t bo vi khun 1.Vch mng phn bo; 2.Ribosom; 3. Mng sinh cht; 4.Vch (thnh t bo); 5.Mc th (mesosom); 6.Nhim sc th; 7.Lng (tin mao); 8.Cht nguyn sinh; 9. V nhy; 10.tim mao(pili chung); 11.pili gii tnh.
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ngha sinh hc ca v nhy l: - Bo v vi khun trong iu kin kh hn, bo v vi khun trnh b thc bo (trng hp ph cu khun-Diplococcus pneumoniae) - Cung cp cht dinh dng cho vi khun khi thiu thc n - L ni tch lu mt s sn phm trao i cht (dextran, xantan...) - Gip vi khun bm vo gi th ( trng hp cc vi khun gy su rng nh Streptococcus salivarrius, Streptococcus mutans...) * Cc loi v nhy hay gip mc (Capsule) - V nhy ln (Macrocapsule) c b dy >0,2 m. V d:Leuconostoc mesenteroides - V nhy b (Microcapsule) c b dy <0,2 m. V d: Pseumococcus denitrificans - Khi nhy ( Zooglea). - C nhng vi khun ch hnh thnh mng nhy trong iu kin nht nh.V d B. anthracis to mng nhy trong mi trng c cha protein ng vt. D.pneumoniae-ch to mng nhy khi xm nhp vo c th ngi hay ng vt. Mun quan st v nhy thng ln tiu bn m vi mc tu, bao nhy c mu trng hin ln trn nn ti. Thnh phn ch yu ca v nhy l polysaccarid, ngoi ra cng c polypeptid v protein. Trong thnh phn polysaccarid ngoi glucose cn c glucosemin, ramnose, acid 2keto-3-deoxygalacturonic, acid uronic, acid pyruvic, acid acetic...
Hnh 3.14. V nhy vi khun Acetobacter xylinum v Leuconostoc mesenteroides Vi khun Acetobacter xylinum c v nhy cu to bi cellulose. Ngi ta dng vi khun Acetobacter xylinum nui cy trn nc da ch to ra Thch da (Nata de coco). Khi vi khun mc trn mi trng c aga (thch 20%), thng to thnh cc dng khun lc: Dng S (Smooth)-trn bng; Dng R(Rough)-x x v dng M(Mucoid)-nhy nht. Hu ht vi khun u c thnh t bo tr Mycoplasma, v Planctomycetales. Chng c cu to t peptidoglycan. Thnh t bo h tr s chuyn ng ca tin mao (flagellum), gip t bo khng vi p sut thm thu, h tr qu trnh phn ct t bo, cn tr s xm
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nhp ca mt s cht c phn t ln, lin quan n tnh khng nguyn, tnh gy bnh, tnh mn cm vi Thc khun th (bacteriophage).
Hnh 3.15. Hnh dng khun lc ca mt s vi khun a. Khun lc ca Staphylococcus aureus; b.Klebsiella pneumoniae ;c.Bacillus subtilis. 4.2. Thnh t bo Nm 1884 Hans Christian Gram ngh ra phng php nhum phn bit phn chia vi khun thnh 2 nhm khc nhau : vi khun Gram dng v vi khun Gram m. Phng php nhum Gram v sau c s dng rng ri khi nh loi vi sinh vt. Thnh phn ho hc ca 2 nhm ny khc nhau ch yu nh sau: Gram dng Thnh phn T l % i vi khi lng Peptidoglycan 30-95 Acid teicoic (Teichoic acid) Cao Lipid Hu nh khng c Protein Khng c hoc c t Gram m kh ca thnh t bo 5-20 0 20 Cao
Hnh 3.16. Thnh t bo vi khun Gram dng
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Hnh 3.17. Thnh t bo vi khun Gram m 4.3. Mng sinh cht Mng sinh cht hay Mng t bo cht (Cytoplasmic membrane, CM) vi khun cng tng t nh cc sinh vt khc. Chng cu to bi 2 lp phospholipid (PL), chim 30-40% khi lng ca mng, v cc protein (nm trong, ngoi hay xen gia mng), chim 60-70% khi lng ca mng. u phosphat ca PL tch in, phn cc, a nc ; ui hydrocarbon khng tch in, khng phn cc, k nc.
Hnh 3.18. Cu trc mng sinh cht Mng sinh cht c cc chc nng ch yu sau y: - Khng ch s qua li ca cc cht dinh dng, cc sn phm trao i cht - Duy tr p sut thm thu bnh thng trong t bo.
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- L ni sinh tng hp cc thnh phn ca thnh t bo v cc polyme ca bao nhy (capsule). - L ni tin hnh qu trnh phosphoryl oxy ho v qu trnh phosphoryl quang hp ( vi khun quang t dng) - L ni tng hp nhiu enzym, cc protein ca chui h hp. - Cung cp nng lng cho s hot ng ca tin mao 4.4. T bo cht T bo cht (TBC-Cytoplasm) l phn vt cht dng keo nm bn trong mng sinh cht, cha ti 80% l nc. Trong t bo cht c protein, acid nucleic, hydrat carbon, lipid, cc ion v c v nhiu nhiu cht khc c khi lng phn t thp. Bo quan ng lu trong TBC l ribosome. Ribosome nm t do trong t bo cht v chim ti 70% trng lng kh ca TBC. Ribosome gm 2 tiu phn (50S v 30S), hai tiu phn ny kt hp vi nhau to thnh ribosome 70S. S l n v Svedberg- i lng o tc lng khi ly tm cao tc.
Hnh 3.19. So snh Ribosome Bacteria v (Eukaryotic ribosome) Trong t bo cht ca vi khun cn c th gp cc cht d tr nh cc ht glycogen, ht PHB (Poly--hydroxybutyrat), Cyanophycin, Phycocyanin, cc ht d nhim sc (metachromatic body), cc git lu hunh... loi vi khun dit cn trng Bacillus thuringiensis v Bacillus sphaericus cn gp tinh th c (parasoral body) hnh qu trm, c bn cht protein v cha nhng c t c th git hi trn 100 loi su hi (tinh th c ch gii phng c t trong mi trng kim do cc vi khun ny hon ton v hi vi ngi, gia sc, gia cm, thu hi sn- c hi i vi tm). Bacillus sphaericus c th dit cung qung ca cc loi mui.
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90
Hnh 3.20. Bo t (spore) v Tinh th c (Crystal) Bacillus thuringiensis (tri) v Bacillus sphaericus (phi). 4.5. Th nhn Th nhn ( Nucleoid) vi khun l dng nhn nguyn thu, cha c mng nhn nn khng c hnh dng c nh, v v vy cn c gi l vng nhn. Khi nhum mu t bo bng thuc nhum Feulgen c th thy th nhn hin mu tm. l 1 nhim sc th (NST, chromosome) duy nht dng vng cha 1 si DNA xon kp ( X khun Streptomyces c th gp nhim sc th dng thng). NST vi khun Escherichia coli di ti 1mm (!), c khi lng phn t l 3.109, cha 4,6.106 cp base nit. Th nhn l b phn cha ng thng tin di truyn ca vi khun.
Hnh 3.21. Th nhn trong t bo vi khun Escherichia coli. Ngoi NST, trong t bo nhiu vi khun cn gp nhng DNA ngoi NST. l nhng DNA xon kp c dng vng khp kn, c kh nng sao chp c lp, chng c tn l Plasmid 4.6. Tin mao v khun mao Tin mao (Lng roi, flagella) khng phi c mt mi vi khun, chng quyt nh kh nng v phng thc di ng ca vi khun. Tin mao l nhng si lng di, di knh hin vi quang hc ch c th thy r khi nhum theo phng php ring. Di knh hin vi in t c th thy rt r cu trc ca tng si tin mao. Mt s vi khun c bao lng (sheath) bao bc sut chiu di si, nh trng hp chi Bdellovibrio hay vi khun t Vibrio cholera. Tin mao ca vi khun c cc loi khc nhau tu tng loi : - Khng c tin mao (v mao, atrichia) - C 1 tin mao mc cc ( n mao, monotricha) - C 1 chm tin mao mc cc ( chm mao, lophotricha)
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- C 2 chm tin mao mc 2 cc ( song chm mao, amphitricha) - C nhiu tin mao mc khp quanh t bo (chu mao, peritricha) C loi tin mao mc gia t bo nh trng hp vi khun Selenomonas ruminantium.
Hnh 3.22. Tin mao vi khun; Cc loi tin mao vi khun Kiu sp xp tin mao lin quan n hnh thc di ng ca vi khun. Tin mao mc cc gip vi khun di ng theo kiu tin- li. Chng o ngc hng bng cch o ngc hng quay ca tin mao. Vi khun chu mao di ng theo hng no th cc tin mao chuyn ng theo hng ngc li. Khi tin mao khng t li v mt hng th vi khun chuyn ng theo kiu nho ln. Tc di chuyn ca vi khun c tin mao thng vo khong 20-80m/giy, ngha l trong 1 giy chuyn ng c mt khong cch ln hn gp 20-80 ln so vi chiu di ca c th chng. Cc chi vi khun thng c tin mao l Vibrio, Spirillum, Pseudomonas, Escherichia, Shigella, Salmonella, Proteus... cc chi Clostridium, Bacterium,Bacillus, ...c loi c tin mao c loi khng. cu khun ch c 1 chi (Planococcus) l c tin mao Xon th c mt dng tin mao c bit gi l tin mao chu cht (periplasmic flagella), hay cn gi l si trc ( axial fibrils), xut pht t cc t bo v qun quang c th. Chng gip xon th chuyn ng c nh s un vn t bo theo kiu vn nt chai.
Hnh 3.23. Xon th (Spirochete) quan st di knh hin vi nn en.
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OS AF PC
Hnh 3.24. C ch chuyn ng un vn t bo Xon th OS: V ngoi, AF: Si trc, PC ng nguyn sinh, IP:L ni 4.7. Khun mao v Khun mao gii: Khun mao (hay Tim mao, Nhung mao , Fimbriae) l nhng si lng rt mnh, rt ngn mc quanh b mt t bo nhiu vi khun Gram m. Chng c ng knh khong 79nm, rng rut (ng knh trong l 2-2,5nm), s lng khong 250-300 si/ vi khun. Kt cu ca khun mao gin n hn nhiu so vi tin mao. Chng c tc dng gip vi khun bm vo gi th ( nhiu vi khun gy bnh dng khun mao bm cht vo mng nhy ca ng h hp, ng tiu ho, ng tit niu ca ngi v ng vt). C mt loi khun mao c bit gi l Khun mao gii (Sex pili, Sex pilus-s nhiu) c th gp mt s vi khun vi s lng ch c 1-10/ vi khun. N c cu to ging khun mao , ng knh khong 9-10nm nhng c th rt di. Chng c th ni lin gia hai vi khun v lm cu ni chuyn vt cht di truyn (DNA) t th cho (donor) sang th nhn (recipient). Qu trnh ny c gi l qu trnh giao phi (mating) hay tip hp (conjugation). Mt s thc khun th (bacteriophage) bm vo cc th th (receptors) khun mao gii v bt u chu trnh pht trin ca chng.
(a) (b) (c) Hnh 3.25. Khun mao (a&b)v (c) khun mao gii (sex pili) vi khun
5. S lc phn loi vi khun

Hnh dng khng cn c coi l mt tiu chun nh danh vi khun, tuy nhin c rt nhiu chi c t tn theo hnh dng (v d nh Bacillus, Streptococcus, Staphylococcus) v n l mt im quan trng nhn dng cc chi ny.
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Mt cng c quan trng nhn dng khc l nhum Gram, t theo tn ca Hans Christian Gram, ngi pht trin k thut ny. Nhum Gram gip phn vi khun thnh 2 nhm, da vo thnh phn cu to ca vch t bo. Khi u tin chnh thc sp xp cc vi khun vo tng ngnh, ngi ta da ch yu vo phn ng ny: Gracilicutes - vi khun c mng t bo th cp cha lipid, nhum Gram m tnh (ni gn l vi khun Gram m) Firmicutes - vi khun c mt mng t bo v vch pepticoglycan dy, nhum Gram cho kt qu dng tnh (Gram dng) Mollicutes - vi khun khng c mng th cp hay vch, nhum Gram m tnh. Cc vi c khun(archeabacteria) trc y c xp trong nhm Mendosicutes. Hu ht vi khun Gram dng c xp vo ngnh Firmicutes v Actinobacteria, l hai ngnh c quan h gn. Tuy nhin, ngnh Firmicutes c nh ngha li v bao gm c mycoplasma (Mollicutes) v mt s vi khun Gram m. Theo quan im hin i (NCBI- National Center for Biotechnology Information, 2005) th vi khun bao gm cc ngnh sau y : 1. -Aquificae 2. -Thermotogae 3. -Thermodesulfobacteria 4. -Deinococcus-Thermus 5. -Chrysiogenetes 6. -Chloroflexi 7. -Nitrospirae 8. -Defferribacteres 9. -Cyanobacteria 10. -Proteobacteria 11. -Firmicutes 12. -Actinobacteria 13. -Planctomycetes 14. -Chlamydiae/Nhm Verrucomicrobia 15. -Spirochaetes 16. -Fibrobacteres /Nhm Acidobacteria 17. -Bacteroidetes/Nhm Chlorobia 18. -Fusobacteria 19. -Dictyoglomi Vic phn ngnh da trn cc c im hnh thi, sinh l, sinh ha, sinh thi... Cn c vo t l G + C trong DNA ngi ta xy dng c cy pht sinh chng loi (Phylogenetic tree) v chia vi khun thnh 12 nhm sau y : 1. 2. 3. -Nhm Oxy ho Hydrogen -Nhm Chu nhit -Nhm Vi khun khng lu hunh mu lc
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4. 5. 6. 7. 8. 9. 10. 11. 12.
-Nhm Deinococcus -Nhm Vi khun lam (Cyanobacteria) -Nhm Proteobacteria -Nhm Chlamydia -Nhm Planctomyces -Nhm Spirochaetes (Xon th) -Nhm Vi khun lu hunh mu lc -Nhm Cytophaga -Nhm Vi khun Gram dng
6. Vi khun c ch v Vi khun gy hi
Vi khun c th c ch hoc c hi cho mi trng, v ng vt, bao gm c con ngi. Vai tr ca vi khun trong gy bnh v truyn bnh rt quan trng. Mt s l tc nhn gy bnh (pathogen) v gy ra bnh un vn (tetanus), st thng hn (typhoid fever), giang mai (syphilis), t (cholera), bnh ly qua thc phm (foodborne illness) v lao (tuberculosis). Nhim khun huyt (sepsis), l hi chng nhim khun ton c th gy sc v gin mch, hay nhim khun khu tr (localized infection), gy ra bi cc vi khun nh streptococcus, staphylococcus, hay nhiu loi Gram m khc. Mt s nhim khun c th lan rng ra khp c th v tr thnh ton thn (systemic). thc vt, vi khun gy mn l (leaf spot), fireblight v ho cy. Cc hnh thc ly nhim gm qua tip xc, khng kh, thc phm, nc v cn trng. K ch (host) b nhim khun c th tr bng thuc khng sinh, c chia lm hai nhm l dit khun (bacteriocide) v km khun (bacteriostasis), vi liu lng m khi phn tn vo dch c th c th tiu dit hoc km hm s pht trin ca vi khun. Cc bin php kh khun c th c thc hin ngn chn s ly lan ca vi khun, v d nh chi da bng cn trc khi tim. Vic v khun cc dng c phu thut v nha khoa c thc hin m bo chng "v khun" (sterile) hay khng mang vi khun gy bnh, ngn chn s nhim khun. Cht ty u c dng dit vi khun hay cc tc nhn gy bnh ngn chn s nhim v nguy c nhim khun. Trong t, cc vi sinh vt sng trong nt r (rhizosphere) bin nitrogen thnh ammoniac bng cc enzym ca chnh mnh. Mt s khc li dng N phn t kh lm ngun nitrogen (m) cho mnh, chuyn nit thnh cc hp cht ca nit, qu trnh ny gi l qu trnh c nh m. Nhiu vi khun c tm thy sng cng sinh trong c th ngi hay cc sinh vt khc. V d nh s hin din ca cc vi khun cng sinh trong rut gi gip ngn cn s pht trin ca cc vi sinh vt c hi. Vi khun c kh nng phn gii cc hp cht hu c mt cch ng kinh ngc. Mt s nhm vi sinh "chuyn ha" ng mt vai tr rt quan trng trong vic hnh thnh cc khong cht t mt s nhm hp cht hu c. V d, s phn gii cellulose, mt trong nhng thnh phn chim a s trong m thc vt, c thc hin ch yu bi cc vi khun hiu kh thuc chi Cytophaga. Kh nng ny cng c con ngi ng dng trong cng nghip v trong ci thin sinh hc (bioremediation). Cc vi khun c kh nng phn hy hydrocarbon trong du m thng c dng lm sch cc vt du loang.
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Vi khun, cng vi nm men v nm mc, c dng ch bin cc thc phm ln men nh fomarge, da chua, nc tng, da ci bp (sauerkraut), dm, ru, v yoghurt. S dng cng ngh sinh hc, cc vi khun c th c "thit k" (bioengineer) sn xut thuc tr bnh nh insulin, hay ci thin sinh hc i vi cc cht thi c hi.
7. Cc vn khc
V mt tin ha hc, vi khun c cho l cc vi sinh vt kh c, xut hin khong 3,7 triu nm trc. T bo eukaryote c cho rng tin ha t s cng sinh ca cc t bo prokaryote. iu r rng l cc bo quan cha DNA nh ty th hay lc lp l nhng phn cng sinh ca mt nhm vi khun hiu kh (ty th) hay vi khun lam (lc lp) c xa. Phn cn li ca t bo c gi thuyt l c ngun gc t t bo archaea. y l ni dung ca thuyt ni cng sinh Tuy nhin, vn cn nhiu tranh ci v ngun gc ca nhng bo quan nh hydrogenosome c phi l ngun gc ca ty th hay ngc li; xem thuyt hydro v ngun gc t bo eukaryote. Vi sinh vt phn b khp mi ni v pht trin nhanh chng nhng ni c thc n, m, v nhit ti u cho s phn chia v ln ln ca chng. Chng c th c mang i bi gi t ni ny sang ni khc. C th ngi l ni c tr ca hng t vi sinh vt; chng trn da, ng rut, trong mi, ming v nhng ni h khc ca c th. Chng c trong khng kh m ta th, nc ta ung v thc n ta n.
III. VI KHUN C BIT

1. X khun (Actinomycetes)
X khun l tn chung ch nhm VSV n bo c hnh si phn nhnh nh nm nhng kch thc v cu trc ging vi vi khun Gram (+). 1.1. c im chung X khun c cu trc si (Hnh 3.29) cho nn t th k 19 tr v trc ngi ta xp nhm x khun vo nm. Khc vi nm x khun c nhn nguyn thy v kch thc nh nh vi khun (b ngang ca si ch khong 0,2-1,0 m). Si x khun c gi l khun ti. X khun phn b rng ri trong t nhin. S lng n v sinh khun lc (CFUcolony-forming unit) x khun trong 1g t thng c 106-108 CFU. Trn mi trng c a s x khun c hai loi khun ti: khun ti kh sinh (aerial mycelium) v khun ti c cht (substrate mycelium). Nhiu loi ch c khun ti c cht nhng cng c loi (nh chi Sporichthya) li ch c khun ti kh sinh. Gia khun lc thng thy c nhiu bo t mng mng gi l bo t trn (conidia hay conidiospores). Nu bo t nm trong bo nang (sporangium) th c gi l nang bo t hay bo t kn (sporangiospores).
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Hnh 3.26. Khun lc v cung sinh bo t Streptomyces sp.
Hnh 3.27. B mt bo t x khun chi Streptomyces (tri), Micromonospora (phi di knh hin vi in t) X khun thuc nhm vi khun Gram dng, thng c t l GC trong DNA cao hn 55%. Trong s khong 1000 chi v 5000 loi sinh vt nhn s cng b c khong 100 chi v 1000 loi x khun. X khun phn b ch yu trong t v ng vai tr rt quan trng trong chu trnh tun hon vt cht trong t nhin. Chng s dng acid humic v cc cht hu c kh phn gii khc trong t. Mc d x khun thuc nhm sinh vt nhn s nhng chng thng sinh trng di dng si v thng to nhiu bo t. Thm ch mt s loi x khun cn hnh thnh ti bo t nh chi Streptosporangium, Micromonospora v bo t di ng nh chi Actinoplanes, Kineosporia. Trc y, v tr phn loi ca X khun lun l cu hi gy nhiu tranh lun gia cc nh Vi sinh vt hc, do n c nhng c im va ging Vi khun va ging Nm. Tuy nhin, n nay, X khun c chng minh l Vi khun vi nhng bng chng sau y: 1. Mt s x khun nh cc loi thuc chi Actinomyces v Nocardia rt ging vi cc loi vi khun thuc chi Lactobacillus v Corynebacterium. 2. X khun ging vi khun ch khng c nhn thc, chng ch cha nhim sc cht phn b dc theo cc si hoc cc t bo. 3. ng knh ca si x khun v bo t ging vi vi khun. ng thi si x khun thng khng cha vch ngn.
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4. X khun l ch tn cng ca cc thc khun th ging nh vi khun, trong khi , nm khng b tn cng bi thc khun th. 5. X khun thng nhy cm vi cc khng sinh c tc dng ln vi khun, nhng li thng khng vi nhng khng sinh tc dng ln nm nh cc polyen. 6. X khun khng cha chitin, cht c mt trong si v bo t ca nhiu nm, m khng c vi khun. ng thi ging nh phn ln vi khun, x khun khng cha cellulose. 7. Tng t vi vi khun, x khun nhy cm vi phn ng acid ca mi trng, c im ny khng c nm. 8. Cc c im v si v nang bo t kn(sporangium) ca chi Actinoplanes cho thy c th chi ny l cu ni gia vi khun v cc nm bc thp. X khun thuc v lp Actinobacteria, b Actinomycetales, bao gm 10 di b, 35 h, 110 chi v 1000 loi. Hin nay, 478 loi c cng b thuc chi Streptomyces v hn 500 loi thuc tt c cc chi cn li v c xp vo nhm x khun him. 1.2. X khun khc nm Kch thc b ngang khun ti nh nh vi khun (0,2-1,0m) Gm 2 loi h si : H si kh sinh v h si c cht. Nhn cha c mng nhn, c kh nng sinh sn nhiu bo t nh nm trn h si kh sinh. Khun lc bm cht vo mi trng thch (dng u que cy khng y i c) c dng i xng ta trn phng x hay ng tm.
Hnh 3.28. X khun Streptomyces scabies (nui trn MT thch nghing v h si) 1.3. Li ch ca x khun - X khun l ngun cung cp cht khng sinh: amphotericin, chloramphenicol, kanamycin, neomycin, streptomycin.. - Ngun sn sinh nhiu enzym. - X khun chi Frankia c kh nng c nh nitrogen thng cng sinh trn r cy phi lao, v nhiu loi cy khc. - Mt s t x khun gy bnh cho ngi, cho vt nui v cho cy trng.
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Hnh 3.29. Nhng bo t v h si ca Frankia u tin c phn lp bi ErnstHeinrich Pommer nm 1959
2. Xon th (Spirochaetales)
Hnh 3.30. Fritz Schaudinn (1871-1906); Xon th Treponema pallidum v ngi b bnh giang mai (Schaudinn v Hoffmann tm ra mm bnh giang mai (Treponema pallidum) nm 1905) Xon th l mt nhm vi khun c bit c t bo di v xon li mc khc nhau. Xon th khng di ng bng lng roi m li di ng bng vic t vn xon c th. B ngang t bo ch vo khong 0,3- 1,5 m, nhng chiu di ty loi m c th thay i t 6500 m. Xon th kh nhum mu, v vy ngi ta thng quan st xon th bng k thut knh hin vi nn en. Khi xon th c mu sng hin rt r trn mt nn ti en. Mt s loi xon th c tr n hi gia, t bo cun xon li xung quanh tr ny. Mt s loi khc li c mng sng trng nh mt gii la mng dnh dc theo t bo. Cc xon th c nhiu ni trong t nhin: ao, h, rut ng vt c v, rut mi g, nhiu loi k sinh trn c th ngi nh Treponema pallidum gy bnh giang mai, Leptospirra australis gy bnh st vng da xut huyt v Borrelia burgdorferi gy bnh st hi quy
3. Rickettsia
Rickettsia c pht hin vo nm 1909 bi nh bc hc M Howard Taylor Ricketts (1871-1910) v Prowazek Stanislaus Joseph Mathias (1875-1915) nh khoa hc Tip Khc. Rickettsia l mt ging ca Rickettsiaceae, Rickettsiales, y l vi khun k sinh ni bo, c 3 nhm (Tribus): Rickettsiae, Ehrlichiae v Wolbachiae vi 2 ging ch yu l Rickettsia v Coxiella, k sinh ng vt c xng. Nhng vi khun ny cng c 2 loi acid
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nucleic, thnh t bo cng c hp cht c trng ca thnh t bo vi khun, sinh sn bng cch chia i khi trong t bo ng vt.
b c Hnh 3.31. a. Howard Taylor Ricketts (1871-1910), b. Vi khun Rickettsia prowazekii v c. K ch Dermacentor andersoni Rickettsia c dng hnh cu, hnh que hoc hnh si. V c i sng k sinh cho nn khng th nui cy c chng trn cc mi trng nhn to m phi nui cy chng trong phi g hay trn c th ng vt (chut, th...). Chng c nng lc khng yu nhit cao, pH nh (pH=4,1), vi formol v nhiu cht st trng khc. Rickettsia l nguyn nhn ca nhiu bnh st pht ban, st ni nt, st phng nc, st chin ho, st Bc c, st Xeberi....Vt truyn bnh c th l chy, rn, b cht, ve...
4. Mycoplasma v Chamydia
Mycoplasma c pht hin vo nm 1898 l loi vi khun nh nht c th nui cy c trn cc mi trng nhn to. Chng nh b n mc c nhiu loi khng th quan st thy c di knh hin vi quang hc. V cha c thnh t bo vng chc nn chng rt d bin i hnh dng. Chng thng c hnh cu, hnh si phn nhnh. Ngi ta thng nui cy Mycoplasma nhit 370C v pH=7,0-8,0. Chng b tiu dit d dng khi a nhit ln 45- 550C trong vng 15 pht. Mycoplasm thng gy ra cc bnh vim phi mng phi, tit niu - sinh dc.
Hnh 3.32. Khun lc v hnh dng t bo Mycoplasma Mycoplasma thuc h Mycoplasmaceae, h c nht ca b Mycoplasmales (i khi l Mycoplasmatales - Freundt, 1955). Trong b Mycoplasmales c cc dng vi khun Mycoplasma dng L v PPLO. y l nhng vi khun thuc Tenerecutes, lp Mollicutes
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- Chlamydia l mt ging ca Chlamydiaceae, thuc b Chlamydiales, l nhng vi khun k sinh bt buc, Gram m v c nhiu im ging Rickettsia, trong ng vt c xng sng chng khng to ra ATP ring m s dng ATP ca vt ch, kch thc ca chng rt nh (0,3 - 0,45 m) v th trong mt thi gian di trc y ngi ta coi chng l mt loi virus (virus kim tnh Van Royen).
Hnh 3.33.Vi khun Chlamydia v bnh au mt ht* do vi khun ny gy nn *Bnh au mt ht (ting Anh: trachoma) l bnh nhim trng mt do vi khun Chlamydia trachomatis c kh nng lm tho, v nu khng cha tr s gy m mt. Bnh mt ht do nhim khun mi mt gy ra, l mt trong nhng nguyn nhn gy m la hng u th gii (c khong 6 triu ngi b m do cn bnh ny). Trong khong 5 - 12 ngy sau khi xm nhp vo mt, vi khun gy vim m v mng ca mt. Mt sng, hng, nga ngy kh chu ri nu lu khng cha s thnh cc vt tho trong m v mt. Khi m mt sng c th lm lng mi qum vo trong, c st vo trng mt to thm vt tho, lm m mt hay m mt. Bnh mt ht l bnh lm m nhng nga cha c hng u trn th gii. Bnh c nhiu ti a phng ngho, chm tin ti Phi chu, Nam , ng Nam v Trung Quc. Mt s cng ng thiu iu kin cng b dch v mt ny, nh th dn c, Nam M v mt s dn o vng Thi Bnh Dng. Lm sng o vim gic mc o ghn o sng m mt, bt m ln thy bn trong c nhiu ht m (do m c tn gi mt ht) o lng mi qum vo trong mt o hch quanh tai o trng en b m iu tr Dng thuc ung khng sinh (Erythromycin, Zithromycin hay Doxycycline) sm c th ngn chn nhng bin chng nh gy tho hay m. Mt vi trng hp cn phu thut ct b tho v sa m mt trnh gy c st lm m mng trng en dn n m. Phng nga V vi khun gy bnh mt ht rt d ly t nc tit ra mi, hng, mt sang thng ngi khc hay qua dng nh khn mt v.v... tng cng gi gn v sinh l cch ngn nga tt nht.
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Cu hi n tp chng 3
a. Chn cu ng nht: 1. c im ca t bo nhn s l: a. T bo cht phn ho cha cc bo quan b. Mng nhn gip trao i cht gia nhn vi t bo cht. c. Cha c mng nhn d. C a,b,c u ng 2. Thnh phn no sau y khng c cu to ca t bo vi khun ? -a. Mng sinh cht; -c.V nhy; -a. V nhy ; -b. Mng li ni cht -d.Lng roi -b. Thnh t bo -d. T bo cht
3. Hnh thi ca vi khun c n nh nh cu trc no sau y: -c. Mng sinh cht;
4. Ngi ta chia lm 2 loi vi khun: vi khun Gram dng v vi khun Gram m da vo yu t no sau y ? -a. Cu trc ca phn t DNA -b. Cu trc ca plasmid -c. Cu trc v thnh phn ha hc ca thnh t bo -d. S lp peptidolycan trong thnh t bo 5. Thnh phn ho hc cu to nn thnh t bo vi khun l: -a. Kitin; -c. Manan; -a. Vi khun lam; -c. To; -b. Cellulose -d. Peptidoglycan -b. Nm men ru -d. ng vt nguyn sinh
6. Sinh vt di y c cu to t bo nhn s l:
7. Trong t bo vi khun ribosome c chc nng no sau y ? -a. Hp thu cc cht dinh dng cho t bo -b. Tin hnh tng hp protein cho t bo -c. Gip trao i cht gia t bo v mi trng sng -d. C 3 chc nng trn. 8. Chc nng di truyn ca vi khun c thc hin bi: -a. Mng sinh cht; -c. Vng nhn ; a. C mng sinh cht -b. T bo cht -d. Ribosome
9. c im cu to t bo nhn s khc vi t bo nhn thc l:
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b. C mng nhn c. C cc bo quan nh b my Golgi, li ni cht, ti th d. Hai cu b v c l ng b. Tr li cc cu hi sau: 1. V s cu to t bo vi khun, trong ghi r cc cu to bt buc phi c v cu to khng thng xuyn ph thuc vo nhm vi khun. 2. So snh cu to thnh t bo vi khun v vi sinh vt c. Ni r vai tr ca thnh trong hot ng sng v trong phng php nhum Gram. 3. V s cu to mng sinh cht ca vi khun, ni r chc nng vn chuyn cc cht qua mng. 4. Bn cht ca cc vt th n nhp, cu to ca chng v kh nng nhum mu. 5. Cht nhn ca vi khun, nhng pht hin mi trong vn genophore ca c th nhn s. 6. Plasmid c th nhn s, vai tr v chc nng. 7. Mng nhy v tin mao, cu to v chc nng cc loi. 8. Ni bo t, cu to v nhum mu. 9. Nu mt s v d vi khun sinh bo t v khng sinh bo t, ng dng ca chng trong cng ngh vi sinh. 10. V s cu to t bo nm men. So snh cu to t bo nm men v nm mc 11. Cc chu trnh sinh hc ca nm men, i din nm mc. 12. Nguyn tc v phng php phn loi vi sinh vt. 13. nh ngha v cho v d cc khi nim sau: bo t v tnh, bo t hu tnh, ni bo t, bo t nh, bo t ti, bo t m, si nm c vch ngn, si cng bo, si hai nhn. 14. So snh tng qut s khc bit ca vi khun, nm, to n bo v ng vt n bo.
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc, NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty, 1997. Vi sinh vt hc, NXBGD, H Ni.
* Ti liu tham kho

1. Dworkin W., Falkow S., Rosenberg E., Schleifer K.-H. & Stackebrandt E. (ed.). 2000. The Prokaryotes: an Evolving electronic resource for the microbiological community. Springer-Verlag, New York. 2. http://vietsciences.net v http://vietsciences.free.fr 3. http://wikipeda
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* Gii thch thut ng

Bacterial mat: Di iu kin thch hp, vi khun c th to thnh nhng khm thy c bng mt thng. C khun (Archaea) l mt nhm vi sinh vt c bit bt ngun t ting La tinh Archaios c ngha l c Gii Archaea Ba ngnh: - Euryarchaeota - Crenarchaeota - Korarchaeota
Vi khun (ting Anh v ting La Tinh l bacterium, s nhiu bacteria) i khi cn c gi l vi trng. Vi khun l mt nhm sinh vt n bo, c kch thc nh (kch thc hin vi) v thng c cu trc t bo n gin khng c nhn, b khung t bo (cytoskeleton) v cc bo quan nh ty th v lc lp. Cu trc t bo ca vi khun c miu t chi tit trong mc sinh vt nhn s v vi khun l sinh vt nhn s, khc vi cc sinh vt c cu trc t bo phc tp hn gi l sinh vt nhn thc. Vi trng l thut ng tng i ph bin vi mt s ngha hiu khc nhau c lit k di y. 1. a s trng hp l tng ng vi vi khun (bacterium/ bacteria), mt nhm sinh vt prokaryote. Ch trng tng ng vi ch khun trong cc cum t ch phn loi hnh thi, vd. "t cu trng" ~ t cu khun, "trc trng" ~ trc khun .v.v. 2. Tng ng vi vi sinh vt trong cm t vi trng hc m ch cc lnh vc nghin cu, b mn, khoa nghin cu trong cc trng i hc v bnh vin, vd. "Khoa Vi trng", "B mn Vi trng hc" .v.v. 3. m ch nhng tc nhn gy bnh, vi ngha rng.
Vi sinh hc
Chng 4: Sinh hc ca cc c th Nhn thc
104
CHNG 4
SINH HC CA CC C TH NHN THC

I. VI NM (MICROFUNGI) *c im c trng Vi nm l g? gm nhng sinh vt nhn thc, h si, phn ln c thnh t bo cha kitin, khng c lc lp, khng c lng v roi. Sng d dng hoi sinh, k sinh, cng sinh, sinh sn ch yu bng bo t. Cc c th nm vi cu to thnh t bo, kiu trao i cht v h enzym khc bit vi cc c th nhn thc khc (thc vt, ng vt) v khc xa vi c th nhn s nn t lu c xp thnh mt gii ring - l gii nm (Kingdom Fungi). C 3 danh t thng dng ch nm l: nm men (Levures, Yeasts), nm mc (Moisissures) v nm ln hay nm c qu th. Trong vi sinh vt hc, vi nm c hiu l cc nm c c th hin vi, bao gm 3 nhm chnh: - Nm n bo hay nm men. - Nm si hay nm mc. - Nm nhy *Ngun gc ca nm Nm c ngun gc a nguyn, cc ngnh, thm ch cc lp trong mt ngnh bt ngun t cc t tin khc nhau. Ngi ta cho rng nm c c t tin t trng roi no . Cc dng nm hin nay xut hin rt lu. Cc bo t ca chng rt ging vi bo t ca nhng loi nm trong lp t ca cc k xa xa. Trong t thuc Mezozoi (cch y 70-185 triu nm) tm thy di tch ca nm rt ging vi nm thuc b Mc nc ca nm non v nm ti cha hon chnh thuc chi Diplodia
1. Nm men (Levures, Yeasts)

1.1. c im hnh thi, cu trc v sinh sn ca nm men Nm men l tn gi thng dng ch mt nhm vi nm c th n bo, nhn c mng nhn, sinh sn ch yu theo kiu ny chi.Nm men khng phi l mt nhm nm ring bit m thuc nhiu nhm khc nhau trong gii nm. Nm men c th thuc v 3 lp nm l nm ti (Ascomycetes), nm m (Basidiomycetes) v nm bt ton (Deuteromycetes). T bo nm men c nhiu hnh dng khc nhau, c th hnh cu, hnh bu dc, hnh elip, hnh ng.... Kch thc t bo thay i t 1,5-12 m. Nu t bo dng si th chiu di c th ti 20 m hay hn na, thng l si nm gi gm nhiu t bo dnh li vi nhau theo chiu di mt cch lng lo. Trong s 75.000 loi nm hin bit th c hn 500 loi nm men thuc khong 50 ging. Thnh t bo nm men thng dy khong 100-250nm, cu to ch yu bng hp cht mannan-glucan hay mannan- kitin. Bn di thnh t bo l mng sinh cht c cu to khng khc g my so vi t bo sinh vt khc.
Vi sinh hc
105
Mng t bo cht nm nm men thng cha 39% lipid, 49% protein, 5%carbonhydrate v 7% nucleic acid. Khc vi vi khun, nm men c nhn phn ha, nhn c mng nhn, l thng v nhn con. Trong t bo nm men c cc c quan nh nh ti th, li ni cht, b my Golgi Ti th nm men c dng hnh cu, hnh bu dc hay hnh si c bao bi hai lp mng, gia hai lp mng l c cht bn lng. T lp mng trong li to ra v s nhng vch ni vo pha trong gi l vch ngang tng din tch b mt ca mng. Ti th l trung tm to nng lng ca t bo. Trong ti th cn gp c mt lng nh DNA, gi l DNA ca ti th. B my Golgi gm nhng tui, nhng khng bo cu to bi cc lp mng xp song song hnh cung. B my Golgi tham gia vo hot ng bi tit cc cht cn b, cc cht c hi ra khi t bo. Li ni cht l h thng cc ng, cc xoang phn nhnh vi cu trc mng tng t nh mng sinh cht. Trn mng li ni cht c rt nhiu ribosome- c quan tng hp ra protein ca t bo.
T bo nm men 1.Thnh t bo, 2.Th bin 3.Mng sinh cht, 4.Nhn t bo, 5.Hch nhn, 6.Mng nhn; 7.Khng bo, 8.Mng li ni cht 9.Ht d tr; 10.Ti th 11.Cht t bo.
Hnh 4.1. S cu to t bo nm men Nm men sinh sn v tnh bng m chi hoc phn chi, gia qu trnh ny c th sinh sn hu tnh. Nm men c 3 dng chu trnh sinh hc: - Chu trnh n bi - lng bi nh loi Saccharomyces cerevisiae. - Chu trnh u th lng bi nh loi Saccharomycodes ludgyzii. - Chu trnh u th n bi nh loi Schizosaccharomyces octosporus. Nh knh hin vi in t khoa hc thy c s khc nhau v thi gian hnh thnh thoi v sc trong sinh sn v tnh nm men phn i v nm men m chi.
Hnh 4.2. Nm men sinh sn bng ny chi
Vi sinh hc
106
Hnh 4.3. Nm men sinh sn bng phn ct t bo bng vch ngn
Bo t ti Bo t mng dy Bo t t Hnh 4.4. Nm men sinh sn bng bo t ti, bo t mng dy v bo t t
Hnh 4.5. Nm men sinh sn bng bo t bn
Hnh 4.6. Bo t ti nm men Schizosaccharomyces
Vi sinh hc
107
Hnh 4.7. S ny chi v hnh thnh bo t ti nm men Saccharomyces cerevisiae 1.2. Vai tr sinh hc ca nm men Nm men c gi tr dinh dng cao v giu protein (45-55% khi lng kh) v vitamin (tin vitamin D v cc loi vitamin nhm B). Nm men li c tc pht trin nhanh, c th ng ha trc tip mui v c (N, P, K) v hu nh cc hp cht carbon hu c u c th (trc tip hoc gin tip) dng lm thc n nui cy nm men (r ng, farafin, du m....) Nm men chim mt v tr c bit trong cng nghip thc phm: lm n bt m, nu ru, lm ru vang, lm pho mt, sn xut sinh khi ch protein. Ring sn xut bnh m hng nm th gii tiu th 1,7 triu tn nm men bnh m. Mt s nm men khc l nguyn nhn gy h hng thc phm. Trong s ny c c nhng loi nm men c th pht trin c c trong mi trng c nng ng rt cao, chng lm hng mt hoa qu, mt ong, xir, mt s khc gy bnh ngi, ng vt v cy trng.
Vi sinh hc
108
Bng 4.1 Bng phn nhm n gin cc nm men

Lp (Classes) B H H ph Ging
Saccharomycetaceae Nm ti (Ascomycetes)
Schizosaccharumvcetoideae Nadsonioideae
Schizosaccharo-myces Hanseniaspora Lipomyces Debaryomyces Hansenula Kluyveromyces Pichia Sacharomyces
Endomycetales
Lipomycetoideae Saccharomycetoi-deae
Spermophthoraceae Nm m (Basidiomycetes) Ustilaginales Tremell-ales Filobasidiaceae Leuvures Sirobasidiaceae Tremellaceae Cryptococcaceae Blastomycetales Cryptococcoideae
Coccidiascus Filobasidium Leucospiridium Sirobasidium Tremalla Brettanomyces Candida Cryptococcus Torulopsis Rhodotoruloideae Trichospororoideae Rhodotorula Trichosporon Sporobolomyces
Nm bt ton (Deuteromycetes)
Sporobolomycetaceae
Hnh 4.8. Hnh thi t bo nm men bnh m Saccharomyces cerevisiae
Candida albicans Cryptococcus neoformans Hnh 4.9. Mt s nm men gy bnh cho ngi
Vi sinh hc
109
2. Nm mc
Nm mc l tn ch cc nhm nm khng phi l nm men, cng khng phi l nm ln c qu th (nm m) 2.1. Cu to ca nm mc Nm mc thng to thnh nhng khi si c nhnh gi l khun ti hay l (si nm).Si nm l mt ng hnh tr di, thng phn nhnh, c loi c vch ngn ngang nh cc lp nm bc cao (Ascomycetes, Basidiomycetes, Deuteromycetes) hoc khng vch ngn thuc v cc nm bc thp nh Oomycetes v Zygomycetes. Mt s loi nm mc thng dng trong thc phm nh Mucor, Rhizopus..., si nm thng khng c vch ngn. Ton b h si c th coi l mt t bo phn nhnh, ngi ta gi y l c th a nhn. Phn ln si nm mc c vch ngn to nn c th a bo. ng knh si nm vo khong 0,3-5 m, cng c khi t ti 10 m hay ln hn (mt thng c th nhn thy). Trn c cht t nhin hoc trn mi trng nui cy c (thch a, thch nghing...), si nm pht trin thnh mt h si (khun ti th) c dng hnh trn v c gi l khun lc nm mc (hnh 2.10)
Hnh 4.10. Khun lc trn mi trng thch a v c quan sinh sn ca nm mc chi Aspergillus Nm mc ch tng trng ngn. iu ng ch l cc vch ngn khng ngn ngang ( cc nm c vch ngn) khng lm cho si nm c cu to a bo hon chnh, v cc vch ngn ny u c l thng (mt hay nhiu l). Qua cc l thng ny sinh cht c th qua li d dng m ngay c nhn cng c th tht nh li chui qua. Kt qu l c nhng t bo c nhiu nhn (thng phn ngn) v ngc li cng c nhng t bo khng c nhn. Ngoi ra trong h si nm, khi c hai u si nm tip cn st vi nhau, th thnh ch tip cn ny c th ha nhy, kt qu l hai si lin thng vi nhau. Chnh v vy, c th ni c h si nm l mt ng thng sut v cht dinh dng cng nh cht nguyn sinh c th lu chuyn d dng trong ng .
Vi sinh hc
110
Hnh 4.11. Hnh thi nm mc Rhizopus Thnh t bo nm mc c cu trc khc nhau ty tng nhm, a s cha kitin, glucan, kitozan...T bo nm mc c cha cc thnh phn tng t nh t bo nm men: Nhn, ti th, b my golgi, li ni cht.
Hnh 4.12. Cu trc i th t bo nm mc 2.2. i sng ca nm mc Nm mc khng cha sc t quang hp, v vy chng ch c th c i sng hoi sinh (trn cht hu c cht), k sinh (trn c th ngi, ng vt, thc vt) hoc cng sinh (vi to trong a y, vi r cy trong nm r)
Vi sinh hc
111
Nm si sinh si ny n bng cch t on si nm, va bng cch to ra rt nhiu bo t v tnh hay hu tnh. Bo t v tnh nh: Bo t kn, bo t mng dy, bo t trn, bo t t, bo t phn.... Bo t hu tnh (to thnh sau qu trnh sinh sn hu tnh)nh: bo t non, bo t tip hp, bo t m...
1 2 3 4 5 Hnh 4.13. Hnh dng ca mt s cung bo t v bo t nm mc thng gp 1.Mucor; 2.Rhizopus; 3. Aspergillus; 4.Penicillium; 5.Fusarium
Thamnidium
Rhizopus
Hnh 4.14. Nm mc sinh sn bng bo t nang kn
Mt s nm c th sinh ra cc ng bo t mt roi (chytridiomycetes) hoc hai roi (Oomycetes) trong chu trnh sinh sn ca mnh. Cc nm bc cao nh loi Aspergillus v Penicillium c th hnh thnh cu tip hp gia hai t bo ca hai si (+ v -), l hin tng sinh sn cn tnh.
Hnh 4.15. Nm mc sinh sn hu tnh bng hnh thc tip hp
Vi sinh hc
112
2.3. Vai tr ca nm mc Nm mc c ng dng trong nhiu ngnh kinh t quc dn. Trong cng nghip dc phm nm mc c s dng sn xut cc thuc khng sinh nh penicillin, cephalosporin, nhiu loi vitamin nh B2...Trong cng nnghip ha cht v cng nghip thc phm, nm mc c s dng sn xut formage, citric acid, nucleic acid, gibberellin, indolacetic acid. Nhiu nm mc c ng dng trong qu trnh chuyn ha cc hp cht steroid v alcaloid (dng lm thuc). Nhiu loi nm si l sinh khi r tin v c th sn xut d dng quy m cng nghip to ra ngun thc n b sung giu protein v vitamin cho ngi, cho gia sc, gia cm. Nm mc thng c nhng enzym phn gii rt mnh nh h enzym phn gii cellulose, phn gii pectin, cc enyme amylase, protease, lipase...T xa xa, nhiu loi nm mc c nhn dn ta s dng sn xut tng, chao, nc chm, to cc enzym... Tuy nhin, c rt nhiu loi nm mc k sinh gy nn nhiu bnh kh cha ngi v gia sc, nhiu loi nm gy tn tht ln lao cho cy trng v cy rng. Cc bnh nm ngi nh hc l, lang ben, nm tc,... la nh o n, m nu, mc sng, kh vn... Bng 4.2. Bng phn loi n gin mt s ging nm mc
Ngnh Ngnh ph
Zygomycotina (Zygomycetes) Ascomycotina
Lp
Zygomycetes Plectomycetes Pyenomycetes
B
Mucorales Eurotiales Spahaeriales Endomycetales Urenidales Ustilaginales
Ging
Mucor Rhizopus Emericella (A. nidulans) Neurospora (N. grassa) Eremothecium Puccinia Ustilago Candida Geotrichum Aspergillus (A. flavus, A. niger) Penicillium P. votatum, P. camenbertii, P. roquefortii)
Amastigomycota
Hemiascomycetes Basidiomycotina (Basidiomycetes) Hemibasidiomycetes
Deuteromycotina (Deuteromycetes)
Hyphomycetes (dng nm Blastomycetes) Hyphomycetes
men
= Moniliales
Vi sinh hc
113
Hnh 4.16. Nm Epidermophyton floccosum gy bnh hc lo ngi
3. Nm nhy
C th nm nhy l mt khi nhy khng c mng gi l th nguyn hnh, thng c mu vng hay hng. Th nguyn hnh c nhiu nhn lng bi, mi nhn l mt t bo, nh vy c th nm nhy l hp bo (plasmodium). Th nguyn hnh c kh nng di chuyn kiu amip. Nm nhy thng sng hoi sinh trn tn tch thc vt,do vy hay gp nm nhy trn bi c, t rng, bi g, cc ng rc hay phn ca ng vt n c. Cng c nhiu loi sng k sinh trn thc vt. Theo Bi Xun ng (1977) ngnh nm nhy (Myxomycetes) chia ra 4 lp: 1. Lp Acrasiomycetes: Gm nm nhy hoi sinh, c ng amip, sau t hp thnh th nhy gi (Pseudoplasopodium). 2. Lp Hydromyxomycetes: Gm nm nhy k sinh c ng amip, sau t hp thnh th nhy gi. 3. Lp Myxomycetes: Hoi sinh c th nhy (plasmodium), bc bo t c cung. 4. Lp Plasmodiophoromycetes: C th nhy, bc bo t khng cung, k sinh l lp nm nhy k sinh gy bnh cho cy trng. Hin nay mt s tc gi cho rng cc Nm nhy c xp vo hai nhm: nhng Nm nhy c cu to cng bo (Myxomycetes) v nhng Nm nhy c cu to t bo (Acrasiomycetes), trc y chng c xp vo Myxomycetes. Trong giai on dinh dng, chng khng c vch t bo v chng hp thu cht dinh dng hay ly thc n theo kiu amip; tng t nh kiu dinh dng ca nguyn sinh ng vt. Tuy nhin, chng thnh lp vch cellulose trong giai on sinh sn, v to bo t c vch bn trong bo t phng, v nh th th ging vi Nm. 3.1. Nm nhy c cu to cng bo Trong giai on dinh dng, Nm nhy c cu to cng bo khng c vch t bo, ch l mt khi cht nguyn sinh trn, cha nhiu nhn c gi l plasmodium. Chng sng trn l cy, g mc hay nhng vt cht hu c khc ly thc n. Di mt s iu kin no nhng th qu rt nh c thnh lp. Loi Physarum polycephalum d nui trong phng th nghim khi cy trn bt la kiu mch. Trong giai on dinh dng, cc plasmodium ny c cc dy cht nguyn sinh cha rt nhiu nhn v cc nhn cng phn chia ng thi. Trc y, chng cn l i
Vi sinh hc
114
tng nghin cu v c im ca cht nguyn sinh, hin nay th chng l i tng nghin cu s nguyn phn. Sau khi bo t phng phng thch cc bo t, chng ny mm to ra nhng th amip n tng, v c th pht trin chin mao. Nhng th amip ny c th hot ng nh nhng giao t, phi hp v bt u cho ra mt plasmodium mi. 3.2. Nm nhy c cu to t bo Nhm ny khc nhm trn l trong giai on dinh dng gm nhng t bo ring l kiu amip, c gi l myxamoeba. Nui cy trong phng th nghim, cc myxamoeba ny n cc vi khun bng cch thc bo. Trong t nhin, Nm nhy c cu to t bo thng c tm thy trn cht hu c thi ra hay phn sc vt. Nhiu loi gii hn s phn b trn phn ca mt loi ng vt c bit m thi. Th d, mt loi ch tm thy trn phn ca kh howler (mt loi kh c ting ku rt ln, sng Nam m), trong khi mt loi khc ch thy sng trn phn ca mt loi i bng. Chng va dinh dng va phn ct gia tng s lng qun th, trc khi bc vo giai on sinh sn k tip. C mt im ng ch l khi mi trng thiu thc n, cc t bo kt dnh li thnh mt plasmodium gi (pseudoplasmodium), v chng phn ng li vi cc kch thch ca mi trng nh mt sinh vt duy nht. V sau plasmodium gi to ra th qu mc thng ln. Trong lc ny, cc myxamoeba phn ha thnh hai loi t bo, t bo cung v bo t (Hnh 4.17).
Hnh 4.17. Chu k i sng ca nm nhy Mt s th nghim cho thy c mt yu t cm ng gip cho cc myxamoeba nhn ra cc t bo cng loi kt li hay nhn ra cc t bo l loi ra. S nhn din ny c s tham gia ca cc glycoprotein hin din trn mng sinh cht ch vo vi giai on ca s pht trin.
Vi sinh hc
115
Tng qut v nm: Bng 4.3. Cc lp nm thng gp

Lp nm Oomycetes Loi si khng c vch ngn (coenocytic) coenocytic c vch ngn, mt s n bo c vch ngn, mt s n bo c vch ngn, mt s n bo Bo t v tnh bo t chuyn ng bo t ti, i khi bo t nh (conidia) nh bo t (m chi) Uredospora, conidia (m chi) bo t nh, bo t t (arthrospora) Bo t hu tnh Oospora Zygospora (bo t tip hp) bo t ti (ascospore) bo t m (basidio-spore) cha thy Ni sng chnh thy sinh, nhiu loi gy bnh cho c, mc sng khoai ty t, phn gii cht hu c thc vt t, phn gii cht hu c thc vt t, phn gii cht hu c thc vt t, thc vt v trn c th ng vt V d Allomyces
Mucor, Rhizopus
Zygomycetes
Ascomycetes
Basidiomycetes
Deuteromycetes
Neurospora, Saccharo-myces, Morchella Agaricus, Amanita Candida, Trychophyton, Epidermophyton
Bng 4.4. Cc kiu bo t nm

V d ging nm Saprolegnia Aspergillus, Penicillium Mucor, Rhizopus Coccidioides Candida Saccharomyces, Neurospora Agaricus Rhizopus Saprolegnia Kiu bo t Bo t v tnh (ng Zoospores) nh bo t (Conidiospore) bo t) Tnh cht Bo t n, c roi, chuyn ng Bo t n hoc tp hp thnh chui c hnh thnh trn cung bo t (Conidiospore) Bo t c hnh thnh trong ti (bo t v tnh) Bo t c hnh thnh bng cch chia t cc si kh sinh Thnh dy, bo t n, c hnh thnh bng phn i hay chi ging nm men c hnh thnh trong ti (bo t hu tnh), thng 4 - 8 bo t trong mt ti Pht trin tn cng ca m (thng 4 bo t) Bo t ln c hnh thnh trong mt thnh dy (bo t hu tnh) Bo t pht trin trong Oogonium (nguyn bo trng)
Bo t ti v tnh (Sporangiospores) Bo t t (Arthrospores) Bo t dy (Chlamydospore), bo t mm (Blastospore) Bo t ti Bo t m (Basidiospores) Bo t tip hp (Zygospores) Bo t non (Oospore)
1 2 3 Hnh 4.18. Mt s loi nm n v nm dc liu c th trng Vit Nam 1. Nm s ( Pleurotus ostreatus ); 2. Mc nh (Auricularia auricula) 3. Nm m (Agaricus bisporus).
Vi sinh hc
116
Cc loi nm m n c nh cc ging nm m (Agaricus bisporus), nm rm (Volvariella volvacea), mc nh (Auricularia), nm hng (Lentinus), ngn nh (Tremella)ang tr thnh i tng ch yu trong cng ngh nui trng nm n. Phn loi nm mc ch yu da vo cc tnh trng hnh thi: cu to si mang bo t, cu to bo t v mt s tnh trng sinh l sinh ha.
II. A Y
a y l dng cng sinh ca hai loi: mt Nm mc v mt To. Hnh thc cng sinh rt c bit, c hnh dng ring nn a y c tn ging v loi. Cc hnh dng ca a y: hnh vy (crustose), cht v dn vo gi th; hnh l vi nhiu thy (foliose) nh l cy; hay hnh cnh (fruticose ), nh bi cy (Hnh 4.19).
Hnh 4.19. Cc dng a y: (A) Dng vy (B) Hnh l (C) Hnh tri cy Trong a y, thnh phn Nm thng l Nm ti, i khi l Nm m. Thnh phn To thng l To lc, i khi l Vi khun lam. T bo To phn tn gia cc khun ty (Hnh 4.20). Mt s khun ty dn cht vo rong hp thu carbohydrat v nhng cht hu c t To, cn Nm th cung cp nc v khong cho To. a y l mt dng thch nghi c bit ca vng kh hn cho php chng c th sng trong nhng iu kin khc nghit.
Vi sinh hc
117
a y hin din trn thn cy, t v . Trn chng l nhng sinh vt tin phong, l nhng tc on u tin chim c mi trng mi v chng c th ph hy dn dn do cc acid m chng tit ra, v s to ra nhng ht t nh. Nhng vt cht hu c t a y thi ra lm tng thnh phn ca t c to ra. Cc acid c tit ra thay i theo loi v thng c dng nh danh a y. a y tng trng vi mt tc rt chm: a y dng vy tng trng t 0, 1 mm n 10 mm /1 nm, a y dng l tng trng t 2 n 4 cm hng nm. a y d b tn hi do cht nhim khng kh v c Hnh 3.20. Cu to ca a y th c xem l sinh vt ch th v cht lng khng kh. Thng phi i xa vi dm cch xa thnh ph mi c th tm c a y. a y ch sinh sn v tnh, mc d thnh phn Nm ti c th sinh sn hu tnh bng ti. Mt mnh ca a y c tch ra cho ra a y mi. Thm vo , mt s loi to ra nhng th sinh sn c bit c gi l mm phn (soredia), l mt khi nh gm cc t bo To c bao quanh bi cc khun ty. Mm phn c pht tn bi gi v nc ma. hiu c bn cht ca a y v gii thch ngun gc ca chng, cc nh thc vt hc t lu th tng hp a y t t bo To v Nm. Mc d c hai thnh phn c nui cy ring r, s t hp li thnh a y tht l kh khn. Trong nhng nm gn y, s cng sinh c to ra, chng c hnh dng phn no ging vi a y nhng cha phi l cu trc tht s ca a y. Do , cu hi c t ra l trong t nhin nhng a y mi t To v Nm c hnh thnh nh th no? a y l thnh phn quan trng ca cc i nguyn vng cc bc v l thc n cho caribou (mt loi nai ln M chu). a y rt quan trng i vi dn Eskimo, dn Lapps, pha bc ca Thy in, Na uy v Phn lan, v caribou l ngun thc phm chnh. Ngoi ra a y cn c dng sn xut nc hoa v phm nhum.
III. VI TO
1. c im chung
Vi to (Microalgae) l tn gi ch tt c cc loi to c kch thc hin vi c sc t quang hp. Chng thuc v nhiu nhm phn loi khc nhau; to mt, to vng nh, to lc, to silic, to gip. Vi to c kh nng sinh sn v tnh ln hu tnh v vi tc ht sc mnh.
Vi sinh hc
118
2. i sng ca vi to
Nhiu loi vi to c i sng tri ni. Chng l thnh phn ch yu ca thc vt ph du. Chng tri ni c trn b mt lp nc v l mt xch thc n u tin ca mi sinh vt sng trong nc. C nhng loi vi to sut i bm vo cc hn , cc cnh cy di y nc. Rt nhiu loi sng trn mt t hoc trong t (to silic, to mt, to lc...). Chng mt mt lm giu thm cht hu c cho t nhng cng c khi tr thnh nhng k tranh dnh cht khong i vi cy trng. Vi to n gin nht l c th n bo, hoc tp hp n bo, c th c roi nh Clamydomonas, Peridinium v Euglena (to mt), hoc khng c roi nh Chlorella (to lc), Diatomia (to silic). Cc vi to thng gp hn l cc c th a bo hoc tp hp n bo, nh cc tp on Volvox, Pediastrum, Scenendesmus (thuc nhm Archethalle) hoc phc tp hn c b phn nh bm v b phn dng ng nh cc si mnh phn nhnh hoc khng (c th c vch ngn to thnh cc t bo tng i c lp hoc khng c vch ngn nh mt ng cng bo (coenocytic). Nhng to ny sinh sn bng cch phn chia nhng t bo l gia hoc bng cch rng t bo u cng (Sphacelaria, Ectocarpus), chng sinh sn hu tnh bng tip hp ng giao (hai giao t bng nhau) hoc d giao (hai giao t khc nhau). Cc sc t quang hp v h tr cc nhm to khc nhau th khc nhau, hin nay ngi ta bit 6 nhm to vi cc sc t c nghin cu tng i k. Bn ging to lc l Clamydomonas (n bo 2 roi), Gonium (tp hp n bo 2 roi), Pandorina (tp hp n bo, pha ngoi cn c 2 roi, pha trong mt roi) v Volvox (tp hp n bo, pha ngoi c 2 roi lm chc nng di ng cho c tp on, pha trong t bo mt roi lm chc nng quang hp, h hp) l mt v d r nt chng minh s tin ha t t chc n bo ln t chc a bo phn ha th s.
Peridinium
Euglena
Ectocarpus
Hnh 4.21. Hnh dng mt s loi vi to
3.Vai tr ca vi to
Li dng tc pht trin c bit nhanh ca cc loi vi to v gi tr dinh dng cao ca chng (giu protein v vitamin) t lu ngi ta tm cch cy nui vi to theo quy trnh cng nghip v th cng nghip. Cc loi vi to c tuyn chn vo sn xut sinh khi thng thuc chi Chlorella v chi Scenedesmus. Ngi ta cho rng trn cc cnh ng v tr dnh cho cc chuyn bay di
Vi sinh hc
119
ngy, cy trng l tng nht chc chn phi l vi to. Kh nng quang hp ca vi to vt gp 5-6 ln so vi cy trng (sau 24 gi c th tng sinh khi ln 1 000 ln). Chng c th thi ra mt lng oxygen mi ngy gp 200 ln th tch ca chng. Ch cn 50 lt dch nui cy to Chlorella c th cung cp oxygen cho mt nh du hnh v tr, mt khc sinh khi Chlorella c th coi l b hn tt c cc loi thc phm thng thng khc. Mt s vi to sng bin c c tnh cao v c th gy cht hng lot ng vt thy sinh.
Hnh 4.22. Hnh thi mt s vi to dng sn xut SPC Chlorella sp. (tri); Spirulina sp. (gia) v Scenedesmus sp. (phi) Bng 4.5. Sc t v mt s tnh cht ca cc nhm to khc nhau
Ngnh to Chlorophy ta (to lc) Dip lc a v b Carotenoid v mt t Oxycaroten Lutein, Zeaxanthine, Neoxanthine, Violaxanthine Astaxanthine, Neoxanthine Mt s tnh cht T bo 2 roi, sinh sn vo tnh bng chia i hoc sinh sn hu tnh, cht d tr l tinh bt, thnh t bo ch yu l cellulose n bo c 1 roi (mt s c 2 - 3 roi), sinh sn v tnh bng chia i hoc hu tnh, cht d tr l m v loi tinh bt paramylon, khng c thnh t bo Phn lp l n bo, mt s nh dng si c 1 - 2 roi, sinh sn v tnh hoc hu tnh, cht d tr l du v lecucosin vi silic, thnh t bo thm pectin, silica n bo 2 roi bn, sinh sn v tnh bng phn i, cht d tr l tinh bt, thnh t bo l cellulosse
Euglenoph yta (to mt)
a v b
Pyrophyta Chrysophy (To la, ta (to vng, to gip) to silic)
a v c
Lutein, Fucoxanthine, Diadinoxanthine, Diatoxanthine Dinoxanthine, Diadinoxanthine, Peridinine
a v c
Phaeophyt a (to nu)
Tucoxanthine, Lutein, a bo, kch thc ln, hai roi khc bit bn, sinh Diatoxanthine, sn v tnh bng ng bo t, sinh sn hu tnh bng Xanthophylls giao t chuyn ng, cht d tr l Laminarian, thnh t bo c cellulosse v acid alginic Phycocyanine, Phycoerythrine, Neoxanthine, Lutein, Zeaxanthine, Violaxanthine Hu ht a bo, kch thc ln, bt ng, sinh sn v tnh bng bo t, sinh sn hu tnh bng giao t, cht d tr l tinh bt, thnh sinh cht ch yu l cellulosse
Rhodophyta (to )
a v d
a v c
Vi sinh hc
120
1. Urothrix
2. Cladophora
3. Vaucheria
6. Stigeoclonium 4. Gonium 5. Pediastrum Hnh 4.23. Mt s loi vi to thng gp
IV. NG VT NGUYN SINH

ng vt nguyn sinh (Protozoa-ting Hy Lp proto=u tin v zoa=ng vt) l nhng sinh vt n bo (nguyn sinh vt-Protista) c kh nng chuyn ng. ng vt nguyn sinh c khong 20,000 n 25,000 loi, trong mt s cng c c kh nng quang hp. ng vt Nguyn sinh l mt dng sng n gin, mc d c th ch c mt t bo, nhng c kh nng thc hin y cc hot ng sng nh mt c th a bo hon chnh, chng c th thu ly thc n, tiu ha, tng hp, h hp, bi tit, iu ha ion v iu ha p sut thm thu, chuyn ng v sinh sn. S d chng c th thc hin c cc hot ng sng l v trong c th cng c nhng cu t ging vi cc cu t t bo ca c th a bo nh nhn, ty th, mng ni cht, h Glogi, khng bo co bp v khng bo tiu ha. Mt s nguyn sinh ng vt cn c bo hu ni lin bo khu vi ti tiu ha, tim mao hoc tin mao hot ng c nh th gc. ng vt nguyn sinh thng c kch thc 0.01 - 0.05mm Nhng ng vt nguyn sinh u tin c Leeuwenhoek A.V pht hin ra ngay t th k XVII nhng c nghin cu vo th k XVIII bi Joblot L. v nhiu tc gi khc. Theo h thng phn loi hin nay th ng vt nguyn sinh c chia lm 4 nhm:
Vi sinh hc
121
1. Sarcomastigophora vi cc nhnh Sarcodina hay Rhizopodes (Rhizopodes v Actinopodes), nhnh Mastigophora hay Flagelles, nhnh Opalinata (cng c ti liu hp nht Rhizopodes v Flagelles vo dng Rhizoflagelles). 2. Sporzoa (k sinh trn ng vt, mt hoc nhiu vt ch). 3. Cnidospora (k sinh trn ng vt c xng v khng xng). 4. Ciliophora hay cilie (roi ngn - cils, c hai loi nhn: nhn to v nhn b). Vi hn 30.000 loi c m t, ng vt nguyn sinh sng t v nc, nhiu ng vt nguyn sinh c vai tr quan trng cc lp bn hot tnh ti cc trm lc nc thi.
1. Vorticella
2. Giardia lamblia 3.Green alga (Chlamydomonas) Hnh 4.24. Mt s loi ng vt nguyn sinh Sau y l so snh mt s tnh cht ca cc nhm vi sinh vt Bng 4.6. So snh mt s tnh cht ca cc nhm vi sinh vt
Vi khun nhn s ha d dng (mt s quang dng) n bo Nm nhn thc ha d dng hu c a bo (tr nm men) ring l, mt s n bo, si khng hnh thnh tp hp vch ngn v si c vch ngn hp th phn bo v t (trc phn) Murein 6,5 - 7,5 k kh n hiu kh cc loi polysaccharide 4000 hp th bo t hu tnh v v tnh Hemicellulose v chitine 3,8 - 5,6 hiu kh glucogen To nhn thc quang t dng mt s n bo v mt s a bo n bo, tp hp si v bt u hnh thnh m quang hp, hp th sc t quang hp v sc t h tr cellulose gn trung tnh hiu kh tinh bt V nguyn sinh nhn thc ha d dng hu c n bo ring l, tp hp
Tnh cht Loi t bo Kiu dinh dng a bo, n bo Cch sp xp t bo Phng php thu nhn thc n Tnh cht c trng Thnh t bo pH ti u Nhu cu O2 Cht d tr chnh
hp th, thc bo chuyn ng khng c hoc lipoproteid trung tnh hiu kh glucogen v nhiu loi polysaccharide 30.000 (ch tnh ng vt nguyn sinh)
S loi hin bit
80.000 (tt c gii nm)
15.000 (ch tnh to n bo)
Vi sinh hc
122
Cu hi n tp chng 4
1. V s cu to t bo vi khun, trong ghi r cc cu to bt buc phi c v cu to khng thng xuyn ph thuc vo nhm vi khun. 2. So snh cu to thnh t bo vi khun v vi sinh vt c. Ni r vai tr ca thnh trong hot ng sng v trong phng php nhum Gram. 3. V s cu to mng sinh cht ca vi khun, ni r chc nng vn chuyn cc cht qua mng. 4. Bn cht ca cc vt th n nhp, cu to ca chng v kh nng nhum mu. 5. Cht nhn ca vi khun, nhng pht hin mi trong vn genophore ca c th nhn s. 6. Plasmid c th nhn s, vai tr v chc nng. 7. Mng nhy v tin mao, cu to v chc nng cc loi. 8. Ni bo t, cu to v nhum mu. 9. Nu mt s v d vi khun sinh bo t v khng sinh bo t, ng dng ca chng trong cng ngh vi sinh. 10. V s cu to t bo nm men. Cu to thnh t bo nm. 11. Cc chu trnh sinh hc ca nm men, i din nm mc. 12. Nguyn tc v phng php phn loi vi sinh vt. 13. nh ngha v cho v d cc khi nim sau: bo t v tnh, bo t hu tnh, ni bo t, bo t nh, bo t ti, bo t m, si nm c vch ngn, si cng bo, si hai nhn. 14. Nu mt s nm c li v gy hi. 15. Cc nhm to, cu to t bo v thnh t bo. 16. ng vt nguyn sinh, cu to c trng khc vi vi khun. So snh tng qut s khc bit ca vi khun, nm, to n bo v ng vt
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc, NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Phm Vn Ty,1999. Vi sinh hc, NXBGD, H Ni.
* Ti liu tham kho

1. Nguyn Thnh t . 1999. Vi sinh vt hc, NXBGD, H Ni. 2. Brock, 2006. Biology of Microbiology 3. http://vietsciences.net v http://vietsciences.free.fr 4. http://wikipedia
Vi sinh hc
123
*Gii ngha t:
Sinh vt nhn thc (eukaryote), cn gi l sinh vt nhn in hnh hoc sinh vt c nhn chnh thc l mt sinh vt gm cc t bo phc tp, trong vt liu di truyn c sp t trong nhn c mng bao bc. Sinh vt nhn thc gm c ng vt, thc vt v nm - hu ht chng l sinh vt a bo - cng nh cc nhm a dng khc c gi chung l nguyn sinh vt (a s l sinh vt n bo). Tri li, cc sinh vt khc, chng hn nh vi khun, khng c nhn v cc cu trc t bo phc tp khc; nhng sinh vt nh th c gi l sinh vt tin nhn hoc sinh vt nhn s (prokaryote). Sinh vt nhn thc c cng mt ngun gc v thng c xp thnh mt siu gii hoc vc (domain). Eukaryote l ch Latinh c ngha l c nhn tht s. Nhn t bo l bo quan ti quan trng trong t bo sinh vt nhn thc. N cha cc nhim sc th ca t bo, l ni din ra qu trnh nhn i DNA v tng hp RNA. V polysaccharide: mt s chng VSV c th to v polysaccharide. V ny cng vi protein A c chc nng bo v vi khun chng li hin tng thc bo. So snh t bo prokaryote v eukaryote - Cu trc quan trng ca t bo prokaryote: mng t bo cht, ribosome v b gen - T bo eukaryote to v phc tp hn nhiu: mng t bo cht, ribosome, nhn t bo cha b gen l nhim sc th, ti th, dip lp S hnh thnh t bo nhn thc - S hnh thnh nhn v h thng phn bo: + T chc b gen theo phng thc phn an qun l dung lng ln thng tin di truyn + m bo s sao m v phn chia trt t b gen t t bo m sang t bo con + To ngun t bin ti t hp phong ph - Ti th: ni cng sinh (endosymbiont) ca t bo tin nhn c phng thc bin dng h hp hiu kh - Lc lp: ni cng sinh ca t bo tin nhn c phng thc bin dng quang nng sinh oxygen.
Vi sinh hc
Chng 5: Dinh dng, sinh trng v sinh sn vi sinh vt
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CHNG 5
DINH DNG, SINH TRNG V SINH SN VI SINH VT

I. THNH PHN HA HC CA T BO VI SINH VT Nhn chung thnh phn t bo VSV cng gm nhng hp cht tng t thnh phn sinh vt bc cao. N thay i ty tng loi, ty tui v mi trng dinh dng. * Cc lin kt ha hc trong t bo VSV -Lin kt cng ha tr - Lin kt hydrogen tng tc k nc -Lc van der Waals * Cc i phn t sinh hc quan trng trong t bo VSV - Protein (55%) - Nucleic acid (23,6%; DNA 3,1%; RNA 20,5%) - Lipid (9,1%), lipopolysaccharide (3,4%) - Polysaccharide (5%) * Thng ngi ta chia cc hp cht trong t bo lm 2 loi: -Phn nc -Phn kh: gm protein, gluxit, lipit, cc nguyn t khong v mt s cht hot ng sinh hc.
1. Nc
Nc c t l ln trong t bo vi khun, n chim t 75- 85% trng lng nguyn sinh cht; cht kh ch chim 15- 25%. Trong t bo vi sinh vt, phn ln nc dng t do, c tc dng ha tan cc cht v c v hu c. Thnh phn nc t do trong t bo c th thay i ty theo iu kin mi trng bn ngoi, trng thi t bo v la tui. S mt nc t do lm ri lon s trao i cht. Ngoi nc t do cn mt phn nc dng lin kt vi cc hp cht th keo ca t bo bng hp ph hoc ni ha hc ta gi l nc cu to. Nc cu to kh tch khi t bo, v khi mt th lm v cu to t bo, t bo s b cht.
2. Protein
Protein l thnh phn ch yu ca phn kh trong t bo vi sinh vt, thng thng chim 80% trng lng kh. Tuy nhin thnh phn protein cao khng phi l c im ca tt c t bo VSV. Mt s VSV ch c 13- 14% protein. S chnh lch ny l do ch mt phn protein trong t bo l protein cu to c vai tr c bn trong qu trnh sng ngoi ra l nhng protein d tr. V lng protein d tr ny nhiu hay t l ty thuc nhiu iu kin, c bit l thnh phn thc n.
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Protein n gin trong VSV bao gm 3 loi: albumin, globulin v glutelin. Protein phc tp l lipoprotein v nucleoprotein, nhiu VSV nucleoprotein chim t 1/3- 1/2 tng s protein. * Cu to protein: Cu to bi chui cc amino acid ni vi nhau bng lin kt peptide - Hai mi amino acid khc nhau v tnh cht ha hc ca cc nhnh bn trong phn t - c tnh rt a dng ca cc protein do trnh t amino acid - Phn t protein c bn cp cu trc: + Cu trc bc mt l trnh t cc amino acid + Cu trc bc hai do s hnh thnh cc vng xon hoc cc phin bn trong si polypeptide do lin kt hydrogen + Cu trc bc ba l cu trc un khc nhiu hn do cc lin kt khng cng ha tr hoc cng ha tr (lin kt SH) + Cu trc bc bn l s kt hp ca nhiu phn t polypeptide
* Nucleic acid
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- DNA v RNA - c to thnh t cc n phn nucleotide - Mt phn t nucleotide gm ng, phosphate v base nitric - DNA v RNA khc nhau thnh phn ng trong nucleotide Lin kt cng ha tr gia cc nhm ng v phosphate ca hai nucleotide k nhau to thnh khung ng phosphate - Trnh t cc base (A, T, G, C, U) trong b khung quyt nh c trng ca phn t nucleic acid - DNA c cu trc mch i gn vi nhau bng lin kt hydrogen gia A - T v G C. Hai mch c trnh t b sung cho nhau. - RNA ch c mch n
3. Gluxit (hay carbonhydrate- saccharide)

Gluxit chim khong 10- 30% cht kh, nm thnh phn ny n 40- 60%. Trong s th 2- 5% l riboza (trong thnh phn axit nucleic). Cn phn ln l nhng polysaccaride thng gp: hemixenlulose, glycogen, glycerin, dextran... Gluxit c gi tr ln trong t bo VSV, n c dng tng hp protein v lipit, lm vt liu xy dng t bo v vt liu nng lng trong qu trnh h hp.
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Polysaccharide VSV - Phn t ng a phn (carbonhydrate, CHO) - Dng n phn cha 4 7 C l ph bin nht trong t bo - n phn thng c sn cu trc chung, khc nhau cc nhm th v v tr khng gian ca nhm OH- trong mch carbon - Lin kt gia cc n phn l lin kt glycoside - Cc polysaccharide khc nhau do khc hng lin kt glycoside (a, b), khc n phn, khc t hp cc loi n phn - Cc polysaccharide quan trng cellulose, glycogen, tinh bt v peptidoglycan.
Hnh 5.1. Cu trc ha hc v mt s dng lin kt glycoside
4. Lipid v cc cht tng t (lipoid):

Lipid rt ph thuc vo thnh phn mi trng thc n. a s VSV t 1- 3% lipid, nhng mt s dng vi khun trong mi trng nhiu gluxit, t hp cht c nit, th lng lipid c th ln ti 50%. Cht bo trong t bo c th dng t do hoc dng kt hp. Cht bo t do c s dng lm vt liu nng lng v cht d tr. - Thnh phn quan trng ca mng
128
- Lipid n gin: triglyceride - Lipid phc tp: c cha P, N, S, cc nhm ng, ethanol amine, serine, choline - Phospholipid quan trng trong cu trc mng
Hnh 5.2. Cu trc ha hc ca mt s dng lipid trong t bo vi sinh vt
5. Mt s cht hu c c hot tnh sinh hc:

Thng chng ch chim mt t l trng lng rt t, nhng chng rt quan trng. ng ch nht l cc enzim, vitamin, sc t, mt s axit hu c, c t, cht khng sinh c tc dng quan trng trong qu trnh trao i cht. Cc cht sinh trng thng bao gm cc vitamin v mt s axit amin c bit. Cc sc t cng thng gp VSV, chnh n mang li cho VSV c mu sc. Mt s sc t ng vai tr rt quan trng trong s ng ha kh CO2.
129
6. Cc nguyn t khong:
Ngoi cc nguyn t hu c C, H, O, N, trong t bo VSV cn c cc nguyn t tro: P, S, K, Ca, Mg, Fe, Na, Mn, v cc nguyn t vi lng B, Mo, Zn, Cu, Br, ... Lng nguyn t tro, v nht l nguyn t vi lng ni chung u t nhng v cng quan trng. Thnh phn cc cht ny cng thay i ty theo c tnh sinh l ca tng loi VSV. V d vi khun lu hunh nhiu S, vi khun st nhiu Fe, vi khun bin nhiu Na v Cl.
II. DINH DNG

1. Cc cht dinh dng
Cc cht dinh dng i vi VSV l nhng cht c VSV hp th t mi trng xung quanh v c chng s dng lm nguyn liu cung cp cho cc qu trnh tng hp to ra cc thnh phn ca t bo hoc cung cp cho cc qu trnh trao i nng lng. Khng phi mi thnh phn ca mi trng nui cy VSV u c coi l cht dinh dng. Cht dinh dng thng l nhng cht c tham gia vo trao i cht ni bo gm: - Cc hp cht c dng lm nguyn liu tng hp nhng vt liu mi ca t bo: Hp cht cha nit to nn nhm amin v imin (NH2, NH). - Cc hp cht cung cp nng lng: ch yu l cc cht cacbon, ng glucose ... - Mui khong: l nhng yu t vi lng nh Ca, P, Mg, Fe, S... Hm lng cht khong thay i tu theo tng loi VSV.
2. Cc loi mi trng sng ca vi sinh vt

2.1. Phn loi da vo thnh phn cht dinh dng a) Mi trng t nhin: gm cc hp cht t nhin, cha xc nh r thnh phn, v d mi trng sa, nc chit tht b, nc chit cc loi rau, qu... nui cy vi khun ngi ta s dng mi trng canh tht c thnh phn nh sau: Tht b- 50g; Peptone trypsine10g, NaCl- 5g; Nc ct 1000 ml. b) Mi trng tng hp hay l mi trng bit thnh phn ho hc. V d mi trng di y dng nui cy Diplococus desulfuricans c thnh phn nh sau: Glucose50g; K2HPO4 1g; NH4Cl 1g; CaSO4 1g; MgSO4.7H2O -2g v nc ct 1000ml. c) Mi trng bn tng hp: trong mi trng ny c cha mt s hp cht t ngun gc t nhin, v mt s cht ho hc bit r thnh phn ho hc, y l loi mi trng hay s dng, v d mi trng EMB: Peptone trypsine-10g; K2HPO4 2g; Eosin 0,4g; Xanh metilen 0,065g; Thch 15g, nc ct 1000ml. d) Mi trng c bit: Dng nui cy mt s VSV c bit c tnh k sinh bt buc. V d: i vi virus gy bnh u ma, ngi ta nui cy chng trn con b cn sng
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v sau thu thp virus trn con b y lm thuc chng bnh u ma. Phn ln cc virus k sinh trn ng vt chng ta phi nui cy trong phi ca trng g ln hoc trn chut, th... 2.2. Phn loi da vo trng thi vt l ca mi trng + Mi trng lng (hay dch th): thnh phn ch c cc cht dinh dng v nc. + Mi trng c (hay cng): l mi trng dch th c b sung 1,5-2% thch (aga). + Mi trng bn lng l mi trng dch th cha 0,35- 0,75% thch. + Mi trng xp l mi trng c cc nguyn liu lm xp (cm , tru, cm, bnh m....) thm dung dch cht dinh dng.
3. Cc kiu dinh dng vi sinh vt

Cn c vo dng nng lng s dng ngi ta c th chia lm cc loi vi sinh vt: - Cc c th quang t dng (photoautotrophic) hay quang dng v c (photolithotrophic) c th chuyn nng lng nh sng thnh nng lng trao i cht (ATP) v cht c tnh kh (NADPH2). CO2 l ngun carbon ch yu, H2O hay H2S l cht cho in t. - C th gp nhng vi khun quang hp thi oxygen ging nh cy xanh, i din l cc loi vi khun lam (Spirulina, Anabaena, Nostoc). Cc to n bo quang hp khng thi oxygen nh cc vi khun lc thuc h Chlorobacteriaceae (Chlorobium, Thiosulfatophilum...), y l nhng vi khun quang dng v c k kh vi H2S hoc S2O32lm cht cho in t. Cc vi khun lu hunh mu ta thuc b Thiorhodobacteriales nh Chromatium, nhng vi khun ny c kh nng pht trin trn mi trng mui v c nh nhiu loi cy xanh, y l nhng vi khun k kh quang dng v c khi c mt trong mi trng S hoc H2S. - Cc vi khun quang d dng (photoheterotrophic) hay quang dng hu c (photoorganotrophic), s dng nh sng lm ngun nng lng v hp cht hu c lm ngun carbon, nh cc vi khun mu ta khng lu hunh thuc b Athiorhodobacteriales, h Athiorhodaceae, cc loi thuc ging Rhodospirillum. Nhng vi khun ny thin v vi hiu kh, l nhng vi khun quang dng hu c, khng c kh nng pht trin vi H2S l cht cho electron c nht, cht cho electron ca chng l Acetate, succinate hay hydro. - Cc vi khun ha dng v c (Chemolithotrophic) hay ha t dng (Chemoautotrophic) s dng nng lng c to thnh t cc phn ng ha hc v CO2 lm ngun carbon. Nng lng c c l nh oxygen ha cc hp cht v c nh NH4+, NO2-, H2, H2S, S, S2O32-, Fe2+... Chng hnh thnh mt nhm rt hn ch tham gia vo cc chu trnh vt cht sng trong t v nc nh Hydrogenomonas oxygen ha hydro, Nitrosomonas oxygen ha NH3, Nitrobacter oxygen ha nitrite, Thiobacillus oxygen ha cc hp cht kh ca lu hunh... - Cc c th ha d dng (Chemoheterotrophic) hay ha dng hu c (Chemoorganotrophic) bao gm cc vi khun, vi nm, ng vt nguyn sinh, ng vt, chng
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s dng nng lng ha hc v mt c cht hu c lm ngun carbon ch yu. Mt nhm ln cc vi khun ha dng hu c l nhng vi sinh vt gy bnh rt c ch trong Y hc, cc vi khun tp nhim vo thc n, cc vi khun c s dng trong cng nghip tng hp cc cht khng sinh, cc loi vitamin, cc amino acid... Da vo ngun thc n carbon kin to nn t bo. Theo , tt c cc VSV c xp vo mt trong nhm sinh l sau:
4. Vi sinh vt nguyn dng v khuyt dng

Ty thuc vo nhu cu ca cc cht ny m ngi ta thng chia VSV thnh hai nhm: - Nguyn dng (prototrophes) l nhng vi sinh vt khng nht thit cn cc nhn t sinh trng, nhng yu t ca mi trng nui cy thng l y di vi chng. - Khuyt dng (auxotrophes) l nhng VSV i hi cc cht hu c nht nh cn cho s sinh trng ca chng.
III. SINH TRNG CA VI SINH VT

1. Cc nhn t sinh trng
Mt s VSV mt kh nng tng hp mt vi hp cht v i hi c cung cp trong mi trng nui cy. l nhng nhn t sinh trng, chng c chia lm 2 loi: - Mt loi cn c cung cp tng lng nh v m nhn nhim v xc tc nh mt thnh phn ca enzym, v d vitamin nhm B; mt loi cn c cung cp vi lng ln hn v c dng lm nguyn liu cu to t bo nh acid amin, purin, pyrimidin. - Mi loi VSV cn nhng nhn t sinh trng khc nhau. Hu nh khng c cht no l cht sinh trng chung i vi cc loi VSV. Th d lin cu khun (Streptococcus) dung huyt nu thiu vitamin B1 s khng nhn ln c, nm men v vi khun lactic cn biotin.
2. iu kin sinh trng

Sinh trng v trao i cht ca vi sinh vt lin quan cht ch vi cc iu kin ca mi trng bn ngoi. Cc iu kin ny bao gm hng lot cc yu t khc nhau, tc ng
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qua li vi nhau. a s cc yu t u c mt c tnh tc dng chung biu hin ba im hot ng l ti thiu, ti thch v cc i. 2.1. C ch tc dng ca cc yu t bn ngoi ln vi sinh vt Cc yu t ca mi trng bn ngoi tc dng ln t bo thuc ba loi l yu t vt l ( m, nhit , p lc, tia bc x...), yu t ha hc (pH mi trng, th oxygen ha kh, cc cht dit khun...) v yu t sinh hc (cht khng sinh, khng th). Tc dng c hi ca cc yu t bn ngoi t bo th hin ch yu nhng bin i sau y: 1. Ph hy thnh t bo: mt s cht nh enzym lysozyme (cha trong l lch, bch cu, lng trng trng, ui thc khun th...) c kh nng phn hu thnh t bo vi khun. 2. Bin i tnh thm ca mng t bo cht 3. Thay i c tnh keo ca nguyn sinh cht: Chng hn nhit cao lm bin tnh protein v lm chng ng t do kh nng kh nc. 4. Km hm hot tnh: mt s cht tc ng vo cc h thng sinh nng lng ca t bo, cyanite km hm cytochrome - oxydase, fluoride ngn cn qu trnh ng phn, cc hp cht ho tr ba ca acsenic bao vy chu trnh Krebs, dinitrophenol km hm qu trnh phosphoryl ho oxygen ha. Cc cht oxygen ha mnh (H2O2, halogen) ph hy cc h thng t bo lm tn hi n chc phn trao i cht. Cc enzym khc c th b bt hot khi lin kt vi cc yu t kim loi nh thy ngn. 5. Hy hoi cc qu trnh tng hp 2.2. Cc nhn t nh hng * Cc yu t vt l - m: Nc cn thit cho hot ng sng ca VSV. Lm mt nc VSV s cht. Tc cht ph thuc vo mi trng ca VSV. Trong hn dch nc thng khi lm mt nc VSV cht nhanh hn l trong hn dch keo. Nu em mi trng lm ng bng trc ri mi lm mt nc th t l cht ca VSV rt thp. Phng php ny c c p dng lm ng kh. Do VSV cn m nht nh sinh trng nn bng cch phi kh hoc sy kh ta c th bo qun c lu di nhiu loi sn phm (hoa qu kh, c, tht kh....). - Nhit : hot ng trao i cht ca vi sinh vt c th coi l kt qu ca cc phn ng ha hc. V cc phn ng ny thuc ph thuc cht ch vo nhit nn yu t nhit r rng nh hng su sc n cc qu trnh sng ca t bo. Nhit thp (di vng sinh ng hc) c th lm bt hot qu trnh vn chuyn cc cht ha tan qua mng t bo cht do thay i cu hnh khng gian ca mt s permease cha trong mng hoc nh hng n vic hnh thnh v tiu th ATP cn cho qu trnh vn chuyn ch ng cc cht dinh dng. - p lc: p sut thm thu v p sut thy tnh c th nh hng n cu trc ca t bo vi khun. a s vi khun sinh trng tt trong mi trng cha t hn 2% mui, nng mui cao hn c hi cho t bo. Nhng cng c mt s vi khun li sinh trng tt nht trong
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mi trng cha ti 30% mui, ta gi l cc vi khun a mui (halophilic). Nhiu vi khun bin thuc nhm ny, chng khng c kh nng pht trin nhng nng ng cao. - Cc tia bc x: Cc tia c th gy nhng bin i ha hc l nh sng mt tri, tia t ngoi, tia X, tia gamma v tia v tr. nh sng mt tri c th gin tip tc ng ln t bo do lm bin i mi trng. Tia t ngoi c tc dng mnh nht bc sng 260nm, y l vng hp th cc i ca acid nucleic v nucleoprotein gy dimer ha thimine, dn n c ch s nhn i ca ADN. Nhng sau khi chiu tia t ngoi, nu a vi khun ra nh sng ban ngy th vi khun c th phc hi kh nng sinh trng v phn chia. Hin tng ny gi l quang ti hot (photoreactivation). - m thanh: sng m thanh, c bit trong vng siu m (trn 20 kHz) c nh hng ln n sinh trng ca vi khun, cc t bo sinh dng b cht nhanh chng. * Cc yu t ha hc - nh hng ca pH mi trng: pH ca mi trng c ngha quyt nh i vi sinh trng ca nhiu vi sinh vt. a s vi khun sinh trng tt nht pH trung tnh (7,0) nh nhiu vi khun gy bnh (mi trng t nhin l mu v bch huyt ca c th ng vt c pH khong 7,4). Cc vi khun nitrate ha, vi khun nt sn, x khun, vi khun phn gii urea li a mi trng hi kim. Mt s vi khun chu acid (vi khun lactic, Acetobacter, Sarcina ventriculi), mt s khc a acid nh Acetobacter acidophilus, Thiobacillus thiooxydans (oxygen ha lu hunh thnh H2SO4) c th sinh trng pH <1. Nm mc v nm men a pH acid (pH 4 - 6). pH mi trng khng nhng nh hng mnh m n sinh trng m cn tc ng su sc n cc qu trnh trao i cht. - nh hng ca oxygen: Tu thuc vo nhu cu i vi oxygen m ngi ta chia vi sinh vt thnh cc nhm sau: + Hiu kh bt buc: thuc nhm ny l cc vi sinh vt ch c th sinh trng c khi c mt oxygen phn t (O2), chng c chui h hp hon chnh, dng O2 lm th nhn hydro cui cng. Trong cc t bo c cha enzym SOD (superoxid dismutase), catalase v peroxidase c tc dng chuyn ho cc gc O 2 v O 2 gii c cho t bo. Tuyt i a s 2 vi nm v s ng vi khun thuc nhm ny. + Hiu kh khng bt buc: thuc nhm ny l cc vi sinh vt c th sinh trng c c trong iu kin c oxygen ln khng c oxygen. Trong t bo c cha SOD v peroxidase, khi c oxygen chng sinh trng tt hn. Phn ln nm men v nhiu vi khun thuc nhm ny. C th k n cc loi nh Saccharomyces cerevisiae, E. coli, Enterobacter aerogenes, Proteus vulgaris. + Vi hiu kh: thuc nhm ny l cc vi sinh vt ch c th sinh trng c iu kin p lc oxygen rt thp. Chng cng thng qua chui h hp v dng oxygen lm th nhn hydro cui cng. C th k n cc loi nh Vibrio cholerae, Hydrogenomonas spp, Zymononas spp, Bacteroides spp... + K kh: vi cc vi sinh vt thuc nhm ny s c mt ca oxygen phn t l c hi. Chng khng sinh trng c trn mi trng c hoc bn c khi trong khng kh hay trong khng kh c cha khong 10% CO2. Chng ch sinh trng c lp th dch su,
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ni khng c oxygen, c cc qu trnh ln men, qu trnh phosphoryl ho quang hp, qu trnh methane ho... Trong t bo ca cc vi sinh vt ny khng c SOD, cytochrome-oxidase, phn ln khng c hydrogenperoxidase. C th k n rt nhiu loi trong cc chi Clostridium, Fusobacterium, Butyrivibrio, Desulfovibrio, Veillonella... - Cc cht dit khun (st trng): thng s dng l phenol v cc dn xut ca phenol, alkol, halogen, kim loi nng, H2O, x phng, thuc nhum v cc cht ty ra tng hp ca cc mui ammon bc bn. * Cc yu t sinh hc Trong cc yu t sinh hc nh hng c hi ln cc qu trnh sng ca vi sinh vt cn k n khng th v khng sinh. Cht khng sinh c th c t nhiu ngun gc khc nhau nh tng hp ho hc, chic xut t thc vt, ng vt nhng ch yu l c tng hp t vi sinh vt. y l cc cht c hiu m nng thp cng c kh nng c ch hoc tiu dit vi sinh vt mt cch chn lc. Khng th l nhng cht c sn trong mu hoc c xut hin khi c khng nguyn xm nhp vo c th, c kh nng lin kt c hiu vi cc khng nguyn lm mt hot lc ca khng nguyn (xem thm phn khng sinh v min dch). 3. Sinh trng ca vi khun trong nui tnh 3.1 Qu trnh sinh trng Nui cy tnh l phng php nui cy m trong sut thi gian ta khng thm vo cht dinh dng cng khng loi b i cc sn phm cui cng ca trao i cht (qun th t bo b gii hn trong mt khong khng gian nht nh). S sinh trng trong mt h thng ng hay h kn nh vy tun theo nhng quy lut bt buc khng nhng i vi cc c th n bo m c i vi cc c th a bo. th biu din s ph thuc ca logarid s lng t bo theo thi gian gi l ng cong sinh trng (hnh 5.3).
Hnh 5.3 : ng cong sinh trng ca vi khun a. Pha lag ; b. Pha log ; c. Pha n nh ; d. Pha t vong
Qu trnh sinh trng ca t bo vi sinh vt trong h kn tri qua cc giai on :
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* Pha lag (pha m u = pha tim tng) Pha ny tnh t lc bt u nui cy n khi vi khun t c tc sinh trng cc i. Trong pha lag vi khun cha phn chia (ngha l cha c kh nng sinh sn) nhng th tch v khi lng t bo tng ln r rt do qu trnh tng hp cc cht, trc ht l cc cao phn t (protein, enzym, acid nucleic...) din ra mnh m. di ca pha lag ph thuc trc ht vo tui ca ng ging v thnh phn mi trng. Thng thng t bo cng gi th pha lag cng di. R rng nguyn nhn ca pha lag l s khc bit gia cc t bo pha n nh (hoc bo t) vi cc t bo ang sinh trng logarid. Trong pha lag din ra vic xy dng li cc t bo ngh thnh cc t bo sinh trng logarid (hoc sinh trng theo ly tha). * Pha logarid (pha log) Trong pha ny vi khun sinh trng v pht trin theo ly tha, ngha l sinh khi v s lng t bo tng theo phng trnh: N = No.2ct hay X= Xo.eut Kch thc ca t bo, thnh phn ha hc, hot tnh sinh l... ni chung khng thay i theo thi gian. T bo trng thi ng hc v c coi nh l nhng t bo tiu chun. * Pha n nh (pha cn bng) Trong pha ny qun th vi khun trng thi cn bng ng hc, s t bo mi sinh ra bng s t bo c cht i. Kt qu l s t bo sng khng tng cng khng gim. Tc sinh trng ph thuc vo nng c cht. Cho nn khi gim nng c cht (trc khi c cht b cn hon ton) tc sinh trng ca vi khun cng gim. Do vic chuyn t pha log sang pha n nh din ra dn dn. Nguyn nhn tn ti ca pha n nh r rng l do s tch ly sn phm c ca trao i c cht (cc loi ru, acid hu c) v vic cn cht dinh dng (thng l cht dinh dng c nng thp nht). Nguyn nhn th nht rt phc tp v kh phn tch, nguyn nhn th hai c nghin cu k hn. * Pha t vong Trong pha ny s lng t bo c kh nng sng gim theo ly tha (mc d s lng t bo tng cng c th khng gim). i khi cc t bo b t phn nh cc enzym ca bn thn. cc vi khun sinh bo t qu trnh phc tp hn do s hnh thnh bo t. Thc ra cha c mt qui lut chung cho pha t vong. S cht ca t bo c th nhanh hay chm, c lin quan n s t phn hay khng t phn. Do sc sng ln bo t b cht chm nht (trong nhng iu kin thch hp nh kh v nhit thp bo t c th c kh nng sng vi trm nm). Nguyn nhn ca pha t vong cha tht r rng, nhng c lin quan n iu kin bt li ca mi trng. Trong trng hp mi trng tch ly cc acid nguyn nhn ca s cht t bo tng i d hiu. Nng cht dinh dng thp di mc cn thit cho hu qu lm gim hot tnh trao i cht, phn hy dn dn cc cht d tr v cui cng dn n s cht ca hng lot t bo. Ngoi c tnh ca bn thn chng vi khun, tnh cht ca cc sn phm trao i cht tch ly li cng nh hng n tin trnh ca pha t vong.
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Mt s enzym th hin hot tnh xc tc cc i trong pha t vong nh deaminase, decarboxylase, cc amylase v protease ngoi bo. Ngoi chc phn xc tc mt s qu trnh tng hp nhng enzym ni trn ch yu xc tc cho qu trnh phn gii. Tc t vong ca t bo c lin quan trc tip n thc tin vi sinh vt hc v k thut, l vn bo qun cc chng vi sinh vt quan trng v mt l thuyt (cc chng v cc bin chng c bit) v k thut (cc chng sinh cht khng sinh, amino acid, vitamin... vi sn lng cao). Ngoi kh nng sng ta cn cn bo qun c cc c tnh di truyn ca vi khun vi nhiu phng php bo qun khc nhau, nhng tt c u nhm lm gim trao i cht n ti thiu ch yu bng cch gim nhit v m Sinh trng kp Khi chuyn cc t bo ang sinh trng logarid vo mi trng mi khc vi mi trng trc ta vn thy c s xut hin pha lag. Nguyn nhn ca pha lag trong trng hp ny chnh l s thch ng ca vi khun vi iu kin nui cy mi; s thch ng c lin quan n vic tng hp cc enzym mi m trc y t bo cha cn. Cc enzym mi ny c tng hp nh s cm ng ca cc c cht mi. V d: Nu chuyn cc t bo ang sinh trng logarid t mi trng khong glucose sang mi trng khong - maltose ta s thy xut hin pha lag, y l thi gian cn cho vic hnh thnh enzym maltase ( - glucozidase). c trng ca hin tng sinh trng kp l ng cong sinh trng gm hai pha lag v hai pha log. Sau khi kt thc pha log th nht t bo li m u pha lag th hai v tip tc pha log th hai (hnh 5.4).
Hnh 5.4: ng cong sinh trng kp E. coli trong mi trng hn hp glucose - sorbitol (bn tri) Aerobacter aerogenes trong mi trng hn hp mui ammon v nitrate (bn phi)
Hin tng sinh trng kp thng gp khi nui cy vi khun trong mi trng cha ngun carbon gm mt hn hp ca hai cht hu c khc nhau. Khi sinh trng t bo s ng ha trc tin ngun carbon no m chng a thch nht. ng thi c cht th nht ny km hm cc enzym cn cho vic ng ha c cht th hai. Ch sau khi ngun carbon th nht cn th ngun carbon th hai mi c th cm ng tng hp nn cc enzym cn trong vic chuyn ha n.
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Hin tng sinh trng kp khng ch hn ch cc ngun carbon v nng lng m thy cc ngun nit v phtpho. Chng hn Aerobacter aerogenes c th s dng ngun nit l amino acid, mui ammon v nitrate. Nu khi cy chuyn vi khun t mi trng nc tht sang mi trng cha mui ammon hoc nitrate th pha lag s biu hin. Nu t mi trng nc tht sang mi trng cha hn hp c hai loi mui s thy c hin tng sinh trng kp. Cc ion NH4+ c ng ho trc ng thi kim ch vic tng hp cm ng enzym nitrate - reductase. Ch sau khi ht mui ammon trong mi trng th mui nitrate mi c s dng. Sinh trng kp l hin tng ph bin v c th gii thch bng c ch kim ch ni chung v c bit bng hiu ng glucose. 4. Cc thng s ca ng cong sinh trng Sn lng t bo c tnh bng hiu gia khi lng cc i (Xmax) v khi lng ban u (X0) khi nui cy vi khun (tnh bng gam): X = Xmax - X0 T s gia sn lng t bo (X) vi lng c cht c dng (S) (tnh bng n v trng lng) gi l h s kinh t (Y). Y=
X S
Nu sn lng tnh bng gam v c cht dng tnh bng mol th gi l h s kinh t mol (Ym). H s kinh t mol (Ym) cho php ta lin h sn lng t bo vi s lng ATP thu c do phn gii mt lng c cht no . Ta c th tnh c sn lng t bo (tnh bng gam) cho 1 mol ATP tiu th. i lng ny gi l h s nng lng (Y ATP). H s nng lng c tnh d dng nu ta bit con ng phn gii ca loi c cht v nng lng ATP sinh ra do s phn gii y. Hng s tc sinh trng ca t bo vi sinh vt trong pha log c tnh theo cng thc: =
ln X t (t ln X 0 t0 ) lg X t lg e (t lg X 0 t0 )
Trong cng thc: X0 v Xt l mt huyn ph t bo ti cc thi im t0 v t, v lge = 0.43429. Thi gian ca pha lag (T1). y l mt thng s quan trng xem xt tnh cht ca vi khun v mi trng nui cy c thch hp hay khng. Thng s ny c xc nh bng hiu gia thi im t r (ti y huyn ph t bo c mt xc nh no x r) v ti (ti y khi t bo c th t n mt m sau nu em nui cy th chng bt u pha log ngay). T1 = tr ti = tn ln x r ln x 0
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V T1 ch thun li so snh 2 ging vi cng tc sinh trng log ging nhau, nn ngi ta thng biu th thi gian ca pha lag khng phi thi gian tuyt i, m l thng qua thi gian sinh l (thi gian ca 1la g). Hiu gia s sinh trng thc t v l thuyt l L = T1 x v. i lng L ch cho bit ging pht trin trong thc t chm hn pht trin theo l thuyt bao nhiu th h ti thi im cui pha lag v chuyn vo u pha log. Thng s ny thng dng so snh cc s liu ni v nh hng ca cc cht dinh dng khc nhau, cc cht c ch sinh trng v iu kin nui cy. 5. Sinh trng ca vi khun trong nui lin tc Trong phng php nui cy tnh ni trn, cc iu kin mi trng lun lun thay i theo thi gian, mt vi khun tng ln cn nng c cht gim xung. Vi khun phi sinh trng v pht trin theo mt s pha nht nh, sinh khi t c khng cao. Tuy nhin trong nhiu nghin cu v thc tin sn xut ta cn cung cp cho vi sinh vt nhng iu kin n nh trong mt thi gian di chng vn c th sinh trng trong pha log. D nhin trong mt mc no c th cy chuyn t bo nhiu ln (qua nhng khong thi gian ngn) vo mi trng dinh dng mi. Nhng n gin hn, ngi ta a mi trng dinh dng mi vo bnh nui cy vi khun ng thi loi khi bnh mt lng tng ng dch vi khun. y chnh l c s ca phng php nui cy lin tc trong chemostas v turbidostas. C s ca phng php nui lin tc: gi s ta c mt bnh nui cy trong vi khun ang sinh trng pht trin. Cho chy lin tc vo bnh mi trng mi c thnh phn khng i. Th tch bnh nui cy c gi khng i, ngha l bnh mi trng i vo c b bi dng mi trng i ra vi cng mt tc . Ta gi th tch bnh l v (lt), tc dng i vo l f (lt/gi). Do tc pha long (cn gi l h s pha long) D = f/v (i lng D biu th s thay i th tch sau mt gi). Nu vi khun khng sinh trng v pht trin, chng s b rt khi bnh nui cy vi th tch: v- = dX = D.X dt
Nh vy mt vi khun trong bnh gim, ta c cng thc tnh: X = X0. e-Dt Tc sinh trng ca qun th vi khun trong bnh biu th bi phng trnh: v+ =
dX = .X dt
t
Nh vy mt vi khun trong bnh tng theo phng trnh: X = X0. e Tc thay i cui cng (tng hoc gim) mt vi khun trong nui cy lin tc l s sai khc gia tng v+ v gim v-:
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v = v+ - v- = Nu Nu
dX = ( - D).X dt
> D => v > 0: mt vi khun trong bnh tng. < D => v < 0: mt vi khun trong bnh gim.
Nu = D => v = 0: mt t bo khng tng khng gim theo thi gian, qun th vi khun trong trng thi cn bng ng hc. Nu bnh th nghim c thit b duy tr lun lun bng D ta s thu c qun th vi khun sinh trng v pht trin theo ly tha, thng xuyn mt mt t bo khng i v khng ph thuc v thi gian. Trong trng hp nh vy khng nhng kch thc trung bnh ca t bo, trng thi sinh l ca chng m c mi trng nui cy u khng i v khng ph thuc vo thi gian. iu ny, mt mt to iu kin cho vic nghin cu sinh trng v sinh l ca t bo vi khun, mt khc ci thin qu trnh sn xut vi sinh vt qui m cng nghip.
Chemostas Turbidostas
Hnh 5.5: Nui cy lin tc trong chemostas (tri) v turbidostas (phi)
kin
Chemostas v turbidostas l hai thit b nui cy trong ta c th duy tr c iu = D.
Nui cy tnh c xem nh h thng ng, qun th t bo sinh trng phi tri qua cc pha m u, logarid, n nh v t vong. Mi pha sinh trng c c trng bi nhng iu kin nht nh. Vic iu khin t ng kh thc hin. Nui cy lin tc, tri li, l h thng m c khuynh hng dn n vic thit lp mt cn bng ng hc. Yu t thi gian y trong phm vi nht nh b loi tr. T bo c cung cp nhng iu kin mi trng khng i, nh vic iu chnh t ng (hnh 5.5).
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IV. S KM HM SINH TRNG V DIT KHUN

1. Cc phng php kh trng
1.1. Kh trng bng nhit - Nhit cao dit t bo vi sinh vt: bin tnh protein, DNA, chc nng mng - Tc cht ph thuc vo nhit v thi gian x l theo mt hm m - Thi gian gim thp phn: khong thi gian lm gim s lng t bo trong qun th theo h s 10-1 - Kh trng bng phng php Pasteur - Kh trng bng cch un si - Kh trng bng phng php hp gin on - Kh trng bng ni hp p lc (Autoclave) - Kh trng bng sc nng kh - Kh trng bng cch t trn ngn la Mun kh trng ta a i a li nhiu ln dng c trn ngn la n cn, bng cch ny mi vi sinh vt bm trn b mt dng c u b tiu dit. 1.2. Kh trng khng bng nhit - Dng nn lc vi khun Mt s cht hu c nh huyt thanh, albumine trong mi trng d b bin tnh khi kh trng bng nhit cao nn phi kh trng bng cch lc qua cc dng c lc vi khun. - Kh trng bng ha cht Cc dung dch st trng nh chloramine 0,5 - 3%, hoc nc phenol 3 - 5%. S dng cn st trng lau cc dng c thy tinh nh que gt, phin knh, l knh, a thu tinh... 1.3. Kh trng bng chiu x - Vi sng, chiu x tia t ngoi, nh sng mt tri, tia X, tia gamma v chm in t. - Tia t ngoi: khng xuyn thu, st trng b mt, khng kh hoc cht lng khng hp th cc bc sng UV - Cc tia gamma, tia X: tnh xuyn thu cao hn, bo qun thc phm, dng c y t
2. Kim sot tng trng ca vi sinh vt bng ha cht

- Cht dit vi sinh vt (cidal agent): cht dit vi khun (bactericidal), cht dit nm (fungicidal), cht dit virt (viricidal) - Cht c ch vi sinh vt (static agent): cht c ch vi khun (bacteriostatic), cht c ch nm (fungistatic), cht c ch virt (viristatic) - Cht st khun (disinfectant) dng cho vt liu khng sng - Cht khng khun (antiseptic) dng trn m sng - Nng c ch ti thiu (minimum inhibitory concentration, MIC)
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Hnh 5.6. Kh trng bng chiu x tia t ngoi
3. Cc phng php bo qun

1. Cy truyn thng xuyn trn thch nghing hoc trch su vo thch. Sau khi sinh trng vi khun c gi trong t lnh +40C. Phng php ny n gin nht v thng c dng nhng km hiu qu nht. 2. Bo qun di du v trng: du paraffin va ngn cn mi trng kh va lm gim trao i cht gy cn tr s xm nhp ca oxygen. 3. Bo qun trong ct hoc t st v trng: do cu trc l - ha ct v t st u l nhng vt cht tt mang cc t bo vi sinh vt, ch yu l cc bo t. 4. ng kh: l phng php hon thin v c hiu qu nht. Vi khun c trn vi mi trng thch hp (sa, huyt thanh...) ri lm lnh v lm kh nh bng kh. 5. Bo qun trong glycerol (10%) v gi trong t lnh su (-600C hay - 800C): y l phng php rt thch hp nhng cn mua c loi ng nha chu nhit (khi kh trng).
V. SINH SN VI SINH VT
1. Sinh sn vi khun (Bacteria)v c khun (Archaea)
- Sinh sn phn i: Vi khun v vi sinh vt c sinh sn bng cch chia i, iu kin thch hp chng pht trin, th tch ln dn, t bo tht li gia, nhn chia lm i, ri cht t bo chia to thnh hai t bo. - Sinh sn bng bo t ny chi: mt s vi khun cn c hnh thc sinh sn bng ngoi bo t (exospora) nh vi khun dinh dng metan (Methylosimus) hay bng bo t t nh x khun Streptomyces craterifer. Vi khun quang dng mu (Rhodomicrobium vannielii) c hnh thc phn nhnh v ny chi(Hnh 5.7).
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Tt c cc bo t sinh sn u ch c cc lp mng, khng c v v khng tm thy hp cht canxi dipicolinat.
b c d g Hnh 5.7. Mt s hnh thc sinh sn vi sinh vt nhn s a. Bo t ny chi Rhodomicrobium sp.; b.Bo t ngi ca Micromonospora sp.; c.Bo t t Streptomyces ; d. Sinh sn phn i Bacteria ; g. Sinh sn bng tip hp vi khun E.coli - Sinh sn hu tnh : Vi khun khng c sinh sn hu tnh, nhng bin i di truyn (hay t bin) vn xy ra trong tng t bo vi khun thng qua cc hot ng ti t hp di truyn. Do , tng t nh cc sinh vt bc cao, kt qu cui cng l vi khun cng c c mt t hp cc tnh trng t hai t bo m. C ba kiu ti t hp di truyn c pht hin vi khun: 1. Bin np (transformation): chuyn DNA trn t mt t bo vi khun sang t bo khc thng qua mi trng lng bn ngoi, hin tng ny gm c vi khun cht. 2. Ti np (transduction): chuyn DNA ca virus, vi khun, hay c virus ln vi khun, t mt t bo sang t bo khc thng qua th thc khun (bacteriophage). 3. Tip hp (conjugation): chuyn DNA t vi khun ny sang vi khun khc thng qua cu trc protein gi l pilus (lng gii tnh). Vi khun, sau khi nhn c DNA t mt trong nhng cch trn, s tin hnh phn chia v truyn b gene ti t hp cho th h sau. Nhiu vi khun cn c plasmid cha DNA nm ngoi nhim sc th (extrachromosomal DNA). Di iu kin thch hp, vi khun c th to thnh nhng khm thy c bng mt thng, chng hn nh bacterial mat.
2. Sinh sn vi sinh vt nhn thc
Hnh 5.8. Mt s hnh thc sinh sn vi sinh vt nhn thc a. Bo t ny chi S. cerevissiae; b. Phn i Schizosaccharomyces; c. sinh sn hu tnh bng hnh thc tip hp Rhizopus.
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2.1. Sinh sn v tnh - Sinh sn sinh dng: Mt khc si nm t vo mi trng thch hp c th pht trin rt nhanh. Phng php ny c dng cy cc ging nm. - Bng ny chi: t bo chn to thnh nhng chi bn (1 hoc2,3).Chi ln dn v sau thnh mt t bo ring. Kiu sinh sn ny in hnh cho cc loi nm men. - Sinh sn bng bo t: Khc vi ni bo t vi khun, bo t nm l loi t bo sinh sn, rt a dng. Chng c cu to ch yu bi 2 lp mng dy hemicellulose v kitin, khng c acid dipicolinic. - Sinh sn phn i: nh Schizosaccharomyces, to lc n bo (Chlorophyta), to mt (Euglenophyta), trng dy (Paramecium).... 2.2. Sinh sn hu tnh Bng hnh thnh hp t do kt hp 2 t bo vi nhau.
3. Khai thc v phng nga ca con ngi i vi VSV

Tc sinh sn nhanh ca vi sinh vt mang li li ch to ln, ng thi cng gy ra nhiu thm ha cho con ngi. 3.1. Vi vi sinh vt c ch, con ngi s dng - Sn xut sinh khi giu dinh dng (Protein, acid amin) - Sn xut nhiu loi sn phm khc nhau (bia, ru ( ung), dm, bt ngt (gia v), thuc cha bnh (cht khng sinh, vitamin), cht xc tc (enzym amylase, protease) - Ch bin v bo qun mt s thc phm (nh: tng, da, c mui, nem chua, sa chua) cng nh th n gia sc (rau, c chua). 3.2. Vi vi sinh vt c hi: Gy h hng thc phm, cc dng hng ha hoc gy bnh cho ngi, gia sc v cy trng, ngi ta phi tm cch pht hin sm c bin php phng tr, iu tr kp thi.
Cu hi n tp chng 5
1. Cc kiu dinh dng ca vi sinh vt? 2. Cc c cht dinh dng cn thit cho hot ng sng ca vi sinh vt? 3. C ch v tc dng ca cc yu t bn ngoi ln sinh trng ca vi sinh vt? 4. Cc pha sinh trng ca vi sinh vt, im khc bit ca phng php nui cy tnh v nui cy lin tc? 5. ngha ca vic nghin cu ng cong sinh trng, hin tng sinh trng kp?
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc, NXB i hc Hu.
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* Ti liu tham kho

1. Nguyn Thnh t, 1999. Vi sinh vt hc, NXBGD, H Ni. 2. http://vietsciences.net v http://vietsciences.free.fr
* Gii thch thut ng

Bioreactor: L phn ng sinh hc (ni ln men) Pha: Giai on; Pha ly tha: (pha log) Pha suy vong: S lng t bo cht vt s lng t bo mi c to thnh. Nc l dung mi ca h thng sng - Chim 70 90% trng lng t bo - C tnh phn cc nh, ha tan cc phn t sinh ha phn cc quan trng trong t bo; kt t cc phn t khng phn cc hnh thnh mng ngn cn s di chuyn ca cc phn t phn cc vo hoc ra khi t bo - L mi trng cho cc phn ng sinh ha trong t bo Tng trng ca qun th vi sinh vt - T bo phn i - S lng t bo s tng theo thi gian v sinh khi s c tng hp vi vn tc tng dn khi qun th tng trng - T bo tng trng vi vn tc khng i nhng s lng t bo tng ln theo hm m (tng trng hm m) N = No 2n - Thi gian gia hai ln phn i c gi l thi gian th h (generation time) - Bin thin ca s lng t bo theo thi gian c biu din bng th tng quan gia hm logarith ca s t bo theo thi gian nui. o c s tng trng - m trc tip bng knh hin vi - m s t bo sng (m khun lc) - Cn sinh khi - o c - MPN (Most Probable Number) c tnh tng trng ca qun th vi sinh vt trong nui cy m -Chu k 4 pha: + pha tim tng (lag phase) + pha hm m (exponential phase) + pha n nh (stationary phase) + pha cht-suy tn (death phase)
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- Cht dinh dng nh hng ng thi n tc t tng trng v hiu sut tng trng (mt t bo, sinh khi) nng thp; ch nh hng n hiu sut tng trng nng cao c tnh tng trng ca qun th vi sinh vt trong nui cy lin tc - H n ha (chemostat) - Tc pha long: thi gian cn b sung thay mi 100% mi trng - c im: pha hm m ko di; trng thi n nh nng ca cht dinh dng gii hn v s lng t bo khng thay i theo thi gian nh hng ca cht dinh dng ln s tng trng ca qun th vi sinh vt trong nui cy lin tc - nh hng n tc tng trng (growth rate): + Tc tng trng thay i theo tc pha long + Khi lng cht dinh dng c thu nhn vo t bo khng p ng c nhu cu tng trng ca t bo - nh hng n nng sut tng trng (growth yield): + Nng sut tng trng thay i theo nng cht dinh dng gii hn + Nng cht dinh dng thp lm gim tng sinh khi ca qun th - 1 mole ATP t d ha to ra 9 10g sinh khi kh ca t bo nh hng ca cc yu t mi trng ln s tng trng ca qun th vi sinh vt - Nhit tng gip t bo c th tng trng nhanh hn nhng c gii hn nhit : + Vi sinh vt n ha ; + Vi sinh vt a lnh + Vi sinh vt a nhit; + Vi sinh vt a nhit cc oan - pH - p sut thm thu v nc hot tnh - Oxygen + Hiu kh; + K kh; + K kh ty
Chng 6: Trao i cht vi sinh vt
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CHNG 6
TRAO I CHT VI SINH VT I. NG PHN

ng phn l qu trnh phn hu phn t glucose (C6H12O6) to thnh pyruvic acid v NADH+ H+. im c bit ca ng phn l khng phi phn t glucose t do b phn hu m phn t ng glucose c hot ho bi vic gn gc P vo to dng ng phosphate. Qu trnh ng phn gm 2 giai on vi nhiu phn ng phc tp: - Phn ct phn t glucose thnh 2 phn t triose l AlPG v PDA. - Bin i 2 phn t triose thnh 2 phn t pyruvic acid. Qu trnh ng phn c th tm tt theo s sau:
Hnh 6.1. S ng phn Kt qu ca ng phn c th tm tt l: C6H12O6 + 2NAD+ + 2ADP + 2H3PO4 2CH3COCOOH + 2NADH+H+ + 2ATP Trong h hp hiu kh, acid pyruvic tip tc phn hu qua chu trnh Krebs, cn 2NADH+H thc hin chui h hp to H2O. 2NADH+H+ + O2 2NAD+ + 2H2O
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Phn ng ny km theo vic tng hp c 6ATP qua qu trnh phosphoryl ho. Vy kt qu ca ng phn trong h hp hiu kh l: C6H12O6 + O2 ng thi to ra c 8 ATP Trong h hp k kh, 2NADH+H+ s c dng kh CH3COCOOH (trong ln men lactic) hay kh CH3CHO (trong ln men ru) nn khng thc hin chui h hp. Phn ng ln men lactic nh sau: 2CH3COCOOH + 2NADH+H+ C6H12O6 Qu trnh ny ch to ra c 2ATP 2CH3CHOHCOOH + 2NAD+ 2CH3CHOHCOOH Vy kt qu ca ng phn trong h hp k kh l: 2CH3COCOOH + 2H2O
II. CHU TRNH TRICARBOXYLIC ACID (Krebs)

Sn phm ca ng phn l acid pyruvic s c decarboxyl ha to acetyl-CoA v mt phn t CO2. Acetyl-CoA tip tc phn hu qua chu trnh Krebs trong h hp k kh. Qu trnh phn hu acid pyruvic qua chu trnh Krebs c thc hin ti ty th do nhiu h enzym xc tc. Chu trnh xy ra qua 2 giai on: - Phn hu acid pyruvic. Trong qu trnh ny s to ra nhiu coenzym kh (NADH+H+, FADH2). - Thc hin chui h hp qua cc coenzym kh c to ra do phn hu acid pyruvic. Chu trnh Krebs c tm tt qua s sau:
(a) (b) Hnh 6.2. a) Krebs, Sir Hans Adolf (1900-1981); b) S minh ho chu trnh Krebs T phn t acid pyruvic qua chu trnh to ra: CH3COCOOH + 3H2O 3CO2 + 5H2 (4NADPH +H+ + 1FADH2) Cc coenzym (NADH + H+, FADH2) thc hin chui h hp:
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5H2 + 5/2O2 Vy kt qu chu trnh s l: CH3COCOOH + 5/2O2
5H2O 3CO2 + 2H2O
T 1 phn t glucose qua ng phn to ra 2 phn t acid pyruvic vi kt qu phn tch trn. T 2 acid pyruvic qua chu trnh Krebs to ra: 2CH3COCOOH + 5O2 6CO2 + 4H2O Kt hp vi giai on ng phn, ta c kt qu tng qut ca qu trnh phn hu glucose qua h hp hiu kh l: C6H12O6 + 6CO2 6CO2 + 6H2O Trong qu trnh phn hu acid pyruvic qua chu trnh Krebs s to c 4 NADH+H+ v 1 FADH2. Cc coenzym kh ny qua chui h hp s tng hp ATP vi kt qu: - 4NADFH+H+ to ra 12 ATP - 1FADH2 to ra 2ATP - Trong chu trnh to ra 1ATP. Nh vy, phn hu 1 phn t acid pyruvic qua chu trnh Krebs to ra 15ATP. Nu phn hu 2 acid pyruvic s to ra 30ATP, kt hp vi giai on ng phn th phn hu 1 phn t glucose to ra c 38ATP.
III. CHUI H HP V PHOSPHORYL HA

1. Chui h hp
Trong t bo s trao i nng lng lun gn vi phn ng oxygen ho-kh. Trong h thng oxygen ho kh hai phn ng oxygen ho v kh lun i km nhau. AH2 + B
+
A + BH2
Trong t bo phn ng trn xy ra thng cn h thng cc cht truyn in t v H trung gian, l h enzym oxygen ho-kh. Cc enzym ny cng vi c cht hot ng trong mt chui phn ng cht ch chuyn H2 t c cht n O2 to nn chui h hp. Khi u ca chui l c cht dng kh AH2. AH2 lm nhim v l cht cho H2. H2 tch ra t c cht c h thng cc coenzym ca h enzyme oxygen ho-kh vn chuyn n khu cui cng ca chui l O2 kh O2 to phn t H2O. Trong chui h hp in t c chuyn t c cht l cht c nng lng cao nht n oxygen c nng lng thp nht, th oxygen ho cao nht (+0,81V). Gia hai thnh phn trn l cc coenzym c th oxygen ho-kh trung gian, th kh gim dn t c cht n O2. Bi vy chui h hp l qu trnh gii phng nng lng. Nng lng thi ra trong chui h hp c xc nh theo phng trnh: G = -nF. Eo (kCal/mol) Trong : G : mc bin i nng lng ca phn ng oxygen ho-kh
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n: s in t trao i trong phn ng. F: s Faraday (23,06) (hng s Faraday). Eo: chnh lch th oxygen ho-kh ca 2 cht tham gia phn ng. Vi phng trnh trn c th xc nh c nng lng thi ra ca tng phn ng trong chui trn c s th oxygen ho-kh ca cc h xc nh.
2. Phosphoryl ho
Qu trnh tng hp ATP trong t bo l qu trnh phosphoryl ho: ADP + H3PO4 ATP + H2O Phn ng ny i hi nng lng tng ng nng lng ca lin kt cao nng th nht (7,3 Kcalo/mol - trong iu kin chun). Tu ngun nng lng cung cp m c cc hnh thc phosphoryl ho quang ha (xy ra trong quang hp) v phosphoryl ha oxygen ha (xy ra trong h hp). Trong h hp c hai hnh thc tng hp ATP - Phosphoryl ho mc c cht: l qu trnh tng hp ATP nh nng lng thi ra ca phn ng oxygen ho trc tip c cht.
Phosphoryl ho qua chui h hp: l qu trnh tng hp ATP nh nng lng thi ra ca cc phn ng trong chui h hp. Chui h hp xy ra nhiu phn ng, phn ng no tho mn cc iu kin ca qu trnh phosphoryl ho th qu trnh tng hp ATP xy ra . Trong chui h hp c 3 v tr iu kin tng hp ATP. Nh vy, c vn chuyn c H2 t c cht n O2 s to ra c 3 ATP cho t bo.
IV. S VN CHUYN CHT DINH DNG VO T BO

tn ti, sinh trng v pht trin, t bo vi sinh vt phi thng xuyn trao i vt cht v nng lng vi mi trng bn ngoi. Mt mt chng nhn cc cht dinh dng cn
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thit t mi trng, mt khc chng thi ra cc sn phm trao i cht. Gia mi trng xung quanh v mi trng bn trong t bo tn ti mt hng ro thm thu, hng ro ny chnh l mng t bo cht. S vn chuyn cc cht qua mng t bo vi sinh vt tun theo mt trong hai c ch: khuch tn n gin (hay cn gi l vn chuyn th ng hay vn chuyn xui dng) v vn chuyn ch ng (vn chuyn ngc dng). S vn chuyn cc cht nh permease c th l th ng (khng cn nng lng ca t bo) hoc ch ng (cn nng lng). Theo c ch vn chuyn th ng, cht ha tan lin kt thun nghch vo mt v tr c bit trn phn t permease nm bn trong mng (c th cc l ca mng). Phc hp cht ha tan - permease c vn chuyn theo hai pha ca mng nh s chnh lch nng mt cht no , ngha l s vn chuyn din ra theo kiu xui dng. S vn chuyn th ng nh permease c chng minh mt s vi sinh vt. Tuy nhin, vi sinh vt c kh nng tch ly mt s cht vi nng cao hn nhiu so vi nng bn ngoi. Chng hn, nng K+ bn trong mt s t bo vi sinh vt c th ln hn nng bn ngoi hng ngn ln. m bo trung ha in, t bo cng ng thi thi ra bn ngoi cc ion H+ hoc Na+. Thm vo , ngi ta pht hin thy cc mng vi khun c hot tnh ca ATP - ase l enzym c lin quan n vic vn chuyn cc cht. R rng, trong t bo vi sinh vt ngoi c ch vn chuyn th ng cn tn ti c ch vn chuyn ch ng nh permease. S vn chuyn ny c tin hnh ngc vi gradien nng ngha l theo kiu ngc dng, nng lng tiu th c th do ATP cung cp. Cho ti nay ngi ta phn lp c hng lot protein vn chuyn trong cc loi vi sinh vt. Ging nh enzym, chng c tnh c hiu c cht khc nhau. Mt s c tnh c hiu gn nh tuyt i. Chng hn, permease ca galactose E.coli ch vn chuyn galactose. Cc permease ca cc ng v acid amin khc th hin tnh c hiu yu hn i vi cc cht ha tan. iu ng ch l trong vi khun s vn chuyn ch ng ca hu ht cc loi ng ph thuc vo qu trnh phosphoryl ha v h thng enzym phosphotransferase. H thng ny bao gm hai enzym EI, EII v mt protein vn chuyn bn nhit (Hpr: heat stable carried protein) c khi lng phn t thp. Cc thnh phn protein ca h thng ny c thun khit v phn ng din ra theo hai bc. Trc ht EI chuyn phosphate t phosphoenolpyruvate (PEP) n Hpr: Hpr + PEP Hpr - P + Pyruvate Sau EII chuyn phosphate t Hpr - P n C6 ca ng n. EI l chung cho nhiu loi ng nhng EII li c trng cho mi loi ng. Ngha l mt t bin no nh hng n vic tng hp EI s dn n mt kh nng vn chuyn nhiu loi ng. Tri li vi t bin nh th i vi EII ch nh hng n s vn chuyn ca mt loi ng (Hnh 6.3).
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Ngoi
Glucose
Mng EII Hpr
Trong Glucose- 6P PEP
EII glucose
Hpr-P
pyruvic
Hnh 6.3. S vn chuyn ng qua mng t bo vi sinh vt
V. TRAO I CHT V NNG LNG

1. Cc ngun nng lng vi sinh vt
Trong th gii vi sinh vt c s khc bit rt ln v kh nng s dng cc ngun nng lng. Mt s vi khun hiu kh cng ging nh ng vt c kh nng tng hp ATP cho mnh t ng b phn gii theo con ng ng phn (glycolyse) v chu trnh Krebs nh chui h hp trn mng t bo cht (tng t nh chui h hp mng trong ca ty th). Cn nhng vi sinh vt k kh li thu nng lng cho hot ng sng ca mnh nh qu trnh ln men ca cc loi ng hoc nh mt loi chui vn chuyn electron trong c s dng cc hp cht khng phi l oxygen phn t lm cht nhn electron cui cng trong chui vn chuyn. y, chui vn chuyn cng khu tr trn mng t bo cht tng t nh chui vn chuyn nm trong cc ty th. i vi cc c th quang dng th li khc, chng c th thu nhn nng lng ca nh sng. ng v quan im nng lng sinh hc, th ty th, lc lp v t bo vi khun c nhiu nt tng ng. Tt c cc vi khun c trn mng t bo cht mt h enzym ATP synthetase tng t nh ATP - synthetase ca cc ty th v lc lp. Phc hp protein vn chuyn qua mng m bo bin i dng proton thnh nng lng trong mi lin kt phosphate dng ATP. 1.1. Cc c th quang dng (phototroph) v quang tng hp (photosynthesis) Hnh thi ca c th quang dng v quang tng hp gia vi khun, to, thc vt c s khc bit r rng. Mt khc, cu to cc c quan quang hp, sc t quang tng hp, cng khc bit. i vi thc vt, cht cho electron l H2O v qu trnh quang hp gii phng oxygen phn t. i vi vi khun quang hp khng thi oxygen v cht cho electron l nhng hp cht v c (nh H2S, S) hoc hp cht hu c (isopropanol) (xem thm chng X). 1.2. Cc c th ha dng v oxygen ha sinh hc Phn ln vi khun khng c c quan v sc t quang hp. Chng ly nng lng hu ch cho cc qu trnh tng hp ca t bo nh gii phng nng lng trong cc phn ng ha hc.
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S trao i cht ca cc c th ho dng ni chung bao gm 3 giai on: - Phn ct cc i phn t ngoi t bo. - Phn gii cc phn t b to ra cc cht trao i trung gian (pyruvate, acetyl CoA) v to nng lng hu ch (ATP). - Phn gii trit cc cht trao i trung gian thnh CO2 v H2O vi vic to ra lng ln ATP. l nhng phn ng oxygen ha c cht hu c v v c. Qu trnh oxygen ha mt c cht A c th xem l mt chui phn ng m trong c cht A mt i cc eletron ca mnh. C cht A gi l cht cho electron s bin thnh mt sn phm c oxygen ha. Tng t, mt hp cht B khc gi l cht nhn electron s bin thnh sn phm kh. Trong phn ln trng hp, mt hp cht A l mt cht hu c c th xem l cht c hydro ha, nh vy th qu trnh oxygen ha v thc cht l mt qu trnh kh hydro (dehydrogenation), trong khi qu trnh kh l qu trnh hydro ha (hydrogenation), t hp hai qu trnh ny l qu trnh oxygen ha kh (oxygendoreduction).
Trong chui phn ng, vic vn chuyn hydro hay electron t c cht sang cht nhn c thc hin nh hng lot enzym, cc enzym ny to thnh chui vn chuyn in t. Cc cht xc tc thch ng cao vi qu trnh oxygen ho c cht l nhng dehydrogenase. Nhng enzym ny c trng cho mt loi c cht v t hp vi cc coenzym m n ng vai tr l cht nhn hydro t c cht. l cc dn xut ca flavin (FMN, FAD) hoc ca pyridine (NAD, NADP) v nhiu hp cht khc. cc c th hiu kh, s vn chuyn cc electron gia coenzym ca dehydrogenase v oxygen phn t c thc hin nh cc hp cht trung gian ca cytochrome.
2. Cc kiu h hp
Thng thng khi cht nhn electron cui cng l oxygen phn t th gi l qu trnh h hp hiu kh v vi sinh vt thuc dng ny gi l vi sinh vt hiu kh (aerobes). Khi cht nhn electron cui cng l mt c cht khc khng phi l oxygen t do, th ngi ta gi l qu trnh ln men hoc c th l qu trnh h hp k kh v vi sinh vt thc hin cc qu trnh ny gi l cc c th k kh (anaerobes).
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Trong s cc c th k kh, cht nhn electron c th l cht hu c hoc cht v c. Nu cht nhn electron l cht hu c ngi ta gi l ln men, nu cht nhn l electron l cht v c ngi ta gi l h hp k kh (anaerobic respiration). H hp ca mt c th hiu kh l mt qu trnh vn chuyn electron v proton qua mng vi oxygen phn t lm cht nhn electron cui cng, cn h hp k kh dng ch mt qu trnh vn chuyn qua mng ca cc electron v proton n cht nhn electron cui cng l mt cht khc oxygen phn t, nh nitrate trong trng hp h hp nitrate (nitrate respiration), fumarate nh h hp fumarate (fumarate respiration ) v.v... Rt nhiu vi sinh vt c nhiu chui vn chuyn electron c th cng hot ng, hoc c hot ng trong nhng iu kin nui cy nht nh. V d nh E. coli, vi khun ng rut ny c chui vn chuyn electron vi oxygen phn t l cht nhn electron cui cng (chui hot ng nh c th hiu kh ) v chui h hp nitrate hot ng k kh trong mi trng nitrate, mt chui h hp fumarate, hot ng k kh trn mi trng cha c cht hu c m vi khun ny c th ln men. H hp c c trng trc ht l ni khu tr ca chui vn chuyn electron mng t bo cht i vi cc c th nhn s v mng trong ca ty th i vi cc c th nhn thc. H hp cn c trng bn cht ca cht nhn electron cui cng l oxygen phn t. S oxygen ho c cht v ng thi s kh ca oxygen c thc hin nh mt chui phn ng enzym, trong c s tham gia ca dehydrogenase v cc coenzym lin kt vi n. C ch thng thy h hp l con ng ca cc cytochrome gin tip. Ngy nay ngi ta hon ton bit r thnh phn v c ch hot ng ca cc cytochrome ny trong t bo nhn thc v trong mt s c th nhn s. Bn cnh qu trnh phosphoryl oxygen ho c cht, cn tn ti nhng con ng khc, c bit l con ng oxygen ho trc tip nh cc enzym vn chuyn cc electron t c cht n oxygen vi s to thnh H2O2 .
Nhng con ng ny to rt t hoc khng to nng lng hu ch. Mt vi khun khng c enzym trn, nhng con ng phn gii trn buc chng phi sng k kh bt buc, bi v oxygen phn t l tc nhn gy c vi chng. C mt s vi khun chu c hiu kh nh Streptococcus, chng khng c catalase, nhng c cc enzym flavin (NAD - oxydase v NAD - peroxydase) cho php chng thc hin s cn bng trong h hp mng t bo cht. Tu thuc vo s lng v lng cc enzym c th phn gii H2O2 m quyt nh tnh hiu kh hay k kh cc vi khun.
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Ln men ch s c mt ca chui vn chuyn electron nm trong t bo cht, khng c s t ng i qua ca dng electron hoc proton phn bn ny v phn bn kia ca mng t bo (mt dng vn chuyn th cp c thit lp nu cn thit, nh nng lng ca ATP). V phng din trao i nng lng, y cng l mt qu trnh oxygen ha kh, nhng cht nhn electron cui cng l cc hp cht hu c, ch khng phi l cc phn t oxygen. Qu trnh ny cng gii phng nng lng nhng t hn nhiu so vi qu trnh h hp. Cc hp cht cho electron (AH2) v cht nhn electron (B) u l cc hp cht hu c. Vic vn chuyn electron c thc hin nh NAD. y c s khc bit r rt so vi h hp v bn cht ca cht vn chuyn electron v sn phm cui cng... C ch vn chuyn electron trong qu trnh ln men c tm tt nh sau:
3. Nghin cu s trao i nng lng

S khc bit v cc phn ng oxygen ha kh cc nhm vi sinh vt khc nhau l mt tiu chun dng nh loi chng, mt s nh nghin cu xut nhng phng php v mi trng c th nh loi nhanh chng v n gin cc nhm vi sinh vt. 3.1. Nghin cu kiu h hp xc nh kiu h hp ca mt loi vi khun, trong phng th nghim ngi ta s dng mi trng VF (nc tht - gan) ch c v a vo ng nghim, ri cy ging vo su, da vo kh nng pht trin trong cc ng nghim ta bit c l loi h hp g. 3.2. Dng phn ng oxydase Phn ng ny cho php s c mt ca chui h hp ang hot ng vi cytochrome C mng nh TMPD (tetra - methyl - paraphenylene - diamine). Phn ng ny c bit hiu qu i vi vi khun G-, phn ng dng tnh vi cc vi khun hiu kh bt buc (tr Pseudomonas maltophyla v Acinatobacter), phn ng m tnh i vi cc vi khun k kh khng bt buc tr cc Vibrio v Aeromonas. c bit i vi cc vi khun ng rut l c phn ng m tnh mc d vi khun ny c chui h hp vi cc cytochrome. 3.3. Th catalase Enzym catalase thng c cc vi khun G-, tr cc ging Streptococcus v Lactobacilus. Khi c mt H2O2, mt vi git huyn ph vi khun c catalase dng s lm si bt gii phng oxygen phn t. Nu: - Si nhiu bt: l huyn ph vi khun hiu kh - Si t bt: vi khun hiu - k kh - Khng c bt: vi khun k kh
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3.4. Phn ng nitrate reductase S kh nitrate c th c nghin cu trn mi trng c nitrate (mi trng lng hoc c) pht hin s c mt ca nitrite (nh thuc th Griess), c th dn n 2 kt qu theo 2 c ch khc nhau: - Kh nitrate ng ha c xc tc bi nitrate reductase B to nitrite, cht ny c th c chuyn ha tip thnh mui ammon l ngun nit d c c th s dng. - Kh nitrate d ha hay h hp nitrate di nh hng ca nitrate reductase A v cc enzym khc, cc nitrate s c kh hon ton thnh N2O v N2 s c gii phng ngoi t bo. H hp nitrate l loi h hp k kh. 3.5. Nghin cu kiu trao i cht Nghin cu ny c thc hin nh s dng glucose lm c cht, loi ng c dng ph bin nht nhiu loi vi khun. Th nghim cn nui cy 2 iu kin hiu kh v k kh. Sau khi nui t m vi nhit thch hp, c th c 4 trng hp xy ra: - Vi khun ln men khi c ging pht trin trong canh trng v lm acid ha trong 2 ng nghim. - Vi khun oxygen ha khi n ch lm acid ha trong ng nghim iu kin hiu kh. - Vi khun bt hot khi khng thy acid ha trong c 2 ng nghim. - Vi khun kim ha khi thy mi trng b kim ho trong iu kin hiu kh, iu ny c th thy khi dng peptone. 3.6. Kh cc sn phm lu hunh Th nghim kh cc sn phm oxygen ha ca lu hunh (sulfate, sulfite, thiosulfate v.v...) c th thy r nh to ra hydro c lu hunh ha (H2S...) pht hin di dng sulfua kim loi b en (sulfua st hay sulfua ch...). 3.7. o mc h hp H hp k (Respirometre) Warburg l dng c u tin dng xc nh mc s dng oxygen ca t bo sng. N bao gm mt bnh cch thy chu nhit mu cn nghin cu. Mu ny c t trong mt bnh cha ni vi mt p k, bnh cha mu v p k to thnh mt nhnh c c nh chc chn trn ng c mc nc di ng, s di chuyn y l do s giao ng ca mu nghin cu gy nn. Bnh cha mu c ni vi mt nhnh ca bnh cong ca p k v c iu chnh c mt dung tch kh nht nh phn trn ca mu nghin cu. loi b nh hng ca CO2 sinh ra trong qu trnh h hp, ngi ta t mt ng nh gia bnh ng mu nghin cu, trong , cha KOH 10% hp th CO2 do qu trnh h hp sinh ra. S thay i p sut trong p k s c tnh chuyn nh dung tch oxygen tiu th. Vi dng c h hp k ny khng nhng dng o s tiu th oxygen phn t m cn c th nghin cu cc phn ng enzym khc na, trong s thay i kh xut hin do kh carbonic, kh amin ha.
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4. S tch tr v s dng nng lng

4.1. Cc mi lin kt giu nng lng T bo sng tch tr nng lng vo cc hp cht v chng c th c gii phng d dng khi t bo cn. Phn ng ha hc ph bin l thy phn mi lin kt esterphosphate. Hp cht c t bo s dng thun li v thng xuyn nht l ATP. Ngoi ra cn c cc hp cht giu nng lng khc nh cc nucleotide UTP, GTP, CTP, TTP; cc acylphosphate, v d nh 1-3 diphosphoglycerate; cc enolphosphate nh PEP; cc acyl - thio - ester nh CoA - SH.
V d: PEP (G = - 53,5 Kj.mole -1)
+ Cc acyl - thioester: V d: Coenzym A 4.2. Ngun gc hp cht cao nng ATP c th c tng hp bng hai c ch: - Qu trnh phosphoryl ha c cht xy ra trong t bo cht. - Qu trnh phosphoryl ha lin h vi gradien electron v proton xy ra mng t bo cht, y ngi ta thy c qu trnh phosphoryl oxygen ha v phosphoryl quang ha (photophosphorylation). Trong c hai trng hp qu trnh tng hp ATP ph thuc vo lng ADP v phosphate v c. + Qu trnh phosphoryl ha c cht: trong qu trnh ny, mt phosphate v c c lin kt (nh xc tc bi enzym) vi cc sn phm ca s phn gii. Hp cht oxygen ha l cht mang mt mi lin kt phosphate giu nng lng v c th c chuyn n ADP. Kiu hnh thnh ATP ny thng thy s phn gii glucose theo con ng EMP (giai on oxygen ha 3 - phosphoglyceraldehyd thnh acid 1,3- diphosphoglyceric v s bin i hp cht ny thnh acid pyruvic). + Qu trnh phosphoryl ha cng lin h vi gradien proton. C khi vn chuyn electron trong qu trnh h hp (hiu kh hoc k kh) cng nh trong quang hp to ra mt gradien proton quan trng v n l nng lng d tr (c th c so vi mt t in c np sn), nng lng d tr ny c th c thu hi bng cch li cc proton mt bn mng m ngi ta thng so snh nh mt ng c sn sinh ATP.
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5. S trao i carbohydrate
5.1. S phn gii carbohydrate Carbohydrate nu dng hp cht c trng lng phn t ln phi c thu phn thnh cc dng ng n c th i vo cc con ng phn gii ng cung cp nng lng cn thit cho hot ng sng ca c th hoc to ra cc hp cht trung gian cn cho s chuyn ho trao i cht. 5.2. S tng hp carbohydrate Tng t nh cc thc vt bc cao, vi sinh vt c kh nng tng hp cc oligo - v polysaccharide. Lng cc oligo - v polysaccharide ni bo t ti 60% khi lng kh ca t bo, cn polysaccharide ngoi bo c th vt nhiu ln khi lng ca vi sinh vt. Thnh t bo cng cha mt lng ln polysaccharide. Tt c cc oligo v polysaccharide c tng hp bng cch ko di chui saccharide c trc nh vic thm n v monosaccharide (X). n v monosaccharide ny tham gia vo phn ng dng nucleotide - monosaccharide c hot ha, thng l dn xut ca uridine diphosphate (UDP.X) nhng i khi cng vi cc nucleotide purine v pirimidine khc. S tng hp din ra theo phn ng chung nh sau: ...X.X.X.X.X.X + UDP - X = ...X.X.X.X.X.X.X. + UDP (n - nhnh) (n + 1 - nhnh) Trong trng hp polysaccharide bao gm hai loi monosaccharide lin tip (X v Y) phn ng chung xy ra qua hai bc: Bc 1: ...X.Y.X.Y.X.Y + UDP - X = ... X.Y.X.Y.X.Y.X. + UDP Bc 2: ...X.Y.X.Y.X.Y.X + UDP - Y = ... X.Y.X.Y.X.Y.X.Y + UDP Con ng tng hp polysaccharide phn nhnh ta cn bit rt t. Trong dch chit t bo ca mt s loi nm si c cha enzym kitine - sinthetase xc tc phn ng chuyn cc nhnh N - acetylglucosamine t UDP - acetylglucosamine n phn t nhn. Mt cch tng t vic tng hp mannan ca thnh t bo nm men cng bao gm vic chuyn cc nhnh mannose t GDP - mannose n cht nhn. Trong vic tng hp tinh bt Chlorella v glycogen Arthrobacter c s tham gia ca ADP - glucose. - Tng hp glycogen: Khi c iu kin ngoi cnh hn ch s pht trin ca vi khun th chng c th tch ly glycogen nh h enzym trng hp cc glucose. Glycogen tch ly di dng cc ht trong cht ngyn sinh m chng ta c th pht hin bng cch nhum mu vi iode (Hnh 6.8).
Hnh 6.4. S tng hp glycogen
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- Tng hp levan v dextran: Cc levan (poly - 2,6-fructose) v cc dextran (poly -1,6 glucose) vi s phn nhnh - 1,3 hoc -1,4 c tng hp t saccharose theo phn ng sau: n saccharose n saccharose n glucose + (fructose)n (glucose)n + n fructose
S hnh thnh nhng hp cht ny chnh l nguyn nhn ca hin tng nhy nht khi nui cy mt s loi vi khun trn mi trng giu saccharose. Cc hp cht polysaccharide cng vi hp cht nit s hnh thnh nn glycopeptide ca thnh t bo.
6. S trao i protein
6.1. S phn gii protein Khc vi ln men, c cht ca qu trnh thi ra l protein. y l mt trong nhng thnh phn quan trng ca xc b ng vt, thc vt v vi sinh vt. S phn gii cc hp cht hu c cha nit c ngha to ln i vi nng nghip v i vi vng tun hon vt cht trong t nhin. Ngi ta gi qu trnh phn gii ny l qu trnh ammon ha (ammonification). Rt nhiu loi vi sinh vt khc nhau tham gia vo qu trnh ammon ha trong t nhin. ng ch l cc vi sinh vt sau: - Vi khun: Bacillus mycoides, B. subtilis, Proteus vulgais, Pseudomonas aeruginosa, E. coli... - X khun v nm: Streptomyces griseus, S. fradiae, Aspergillus niger, A. awamori, Penicillium camemberti, Mucor spp., ...
Trong c th vi sinh vt, cc acid amin thng c chuyn ha nh qu trnh kh amin, qu trnh kh carboxyl hoc ng thi va kh amin va kh carboxyl. Khi phn gii cc acid amin ty theo tng loi phn ng s to cc sn phm trung gian, y l tin cht cho cc phn ng sinh tng hp hoc cn thit cho mt s qu trnh trao i cht, nh s to thnh acid fumaric t acid aspactic:
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HOOC- CH2- CH (NH2) - COOH HOOC-CH=CH-COOH+ NH3 hay to - ketoglutaric t acid glutamic, acid acetic t glycine, acid succinic t aspactic...). Trong qu trnh kh amin thy phn c s lin kt vi cc ion H+ v OH- nh s to thnh acid lactic t alanine: CH3-CH(NH2)-COOH + H2O CH3-CHOH-COOH + NH3 Nhiu sn phm sinh ra trong qu trnh phn gii acid amin (ru, acid hu c ...) s c li cun vo qu trnh phn gii to nng lng. Nhiu loi amin do vi khun sn sinh ra trong c th ngi v ng vt c tnh c, nh s tch ly histamine c th gy chng co git mch mu, cc hp cht diamine nh acmatine, putrecine gp nhiu trong tht c i thiu u c tnh c. Mt s vi khun c kh nng chuyn ha tryptophan thnh 2 cht c mi hi l indol v skatol. Trong qu trnh phn gii cc acid amin cha lu hunh (methionine, cysteine, cystine) s to ra H2S lm cho phn hoc tht c c mi hi.
Khi qu trnh thi ra xy ra trong iu kin thong kh th qu trnh oxygen ha c tin hnh n cng, khi hu ht carbon c chuyn thnh CO2. Nu qu trnh ny din ra trong iu kin k kh th c s tch ly nhiu sn phm trung gian ni trn. Ngoi kh nng phn gii protein, vi sinh vt cn c kh nng phn gii mt s cht m khc nh urea, acid uric... S phn gii cc cht ny cng to ra cc sn phm trung gian nh acid formic, acid acetic, CO2, NH3..., cc cht ny c th tham gia trc tip vo qu trnh trao i cht v trao i nng lng ca t bo. Urea l cht hu c ch yu trong nc tiu ca ngi v ng. Trong urea c cha 47% nit nhng nu khng c vi sinh vt phn gii thnh NH3 th ngun nit ln lao ny cng tr thnh v ch i vi mi thc vt. Rt nhiu vi khun, x khun, nm si sng trong t c kh nng phn gii mnh m urea. ng ch l cc loi nh Planosarcina urea, Micrococcus urea, Bacillus pasteurii, Proteus vulgaris... Qu trnh phn gii urea xy ra rt n gin vi s xc tc ca urease:
NH3 sinh ra do s phn gii hp cht hu c thng c oxygen ha thnh NO2- v tip tc oxygen ha n NO3-. Phn m ((NH4)2SO4 hoc NH4Cl) sau khi bn rung cng nhanh chng chuyn thnh NO3-. Qu trnh chuyn ho ny gi l qu trnh nitrate ha. Qu trnh ny bao gm 2 giai on:
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- Qu trnh nitrite ha c biu th bng phng trnh phn ng sau:
Nng lng sinh ra trong phn ng ny cng c vi khun nitrite s dng ng ha CO2 ca khng kh. Thc ra qu trnh ny tri qua nhiu phn ng trung gian v to nhiu sn phm trung gian, trong sn phm trung gian quan trng nht l hydroxygenlamin (NH2OH). Enzym xc tc cho vic chuyn hydro l cc enzym ca qu trnh h hp hiu kh. Vi khun nitrite l bn t dng bt buc vi cc chi i din l Nitrosomonas, Nitrosococcus, Nitrosospira v Nitrosocystis. - Qu trnh nitrate ha c biu th bng phng trnh sau:
Cng nh cc vi khun t dng ha nng khc, nng lng sinh trong phn ng oxygen ho NO2- c s dng ng ha CO2 khng kh. Vi khun nitrate ha in hnh l cc loi thuc ging Nitrobacter. y l nhng t bo hnh bu dc, kch thc khong 0,81,0 m, thuc loi G-, khng to bo t, khun lc nh b v trong sut. Ngoi ra cn c mt s ging khc nh Nitrococcus, Nitrospira... 6.2. S tng hp protein Ba qu trnh ch yu trong vic bin i v truyn t thng tin di truyn trong c th l: - S nhn i DNA to ra 2 phn DNA ging nhau v ging vi phn t DNA m. - Qu trnh sao m theo thng tin di truyn t DNA c sao sang RNA, sau c chuyn n ribosome. - Qu trnh dch m cho php chuyn dch thng tin di truyn thnh cc amino acidcu trc nn protein. Cng ngh di truyn cho php gii m trt t cc nucleotide thnh trt t cc amino acid c xc lp (h. 6.5). Phn ln cc vi sinh vt c kh nng tng hp 20 loi amino acid. Con ng tng hp amino acid vi sinh vt c nghin cu kh t m. Nguyn liu dng cho qu trnh tng hp l nhng sn phm trung gian nh pyruvate, - ketoglutarate, oxaloAcetate, fumarate... Trong phn ln trng hp, nhm amin c a vo giai on cui cng ca sinh tng hp nh s chuyn amin, mt s amino acid c hnh thnh nh hng lot bin i ca cc amino acid khc, trong trng hp khng cn s chuyn amin. Nhiu loi L - amino acid c sinh tng hp bng con ng ln men nh vi sinh vt nh lysine, glutamic, alanine, glycine... Hin nay nhiu loi acid amin t nhin l sn phm ca cng ngh sinh hc bng cch thy phn protein, tng hp ha hc hoc sinh tng hp nh vi sinh vt. Trong nhiu trng hp ngi ta phi hp gia tng hp ha hc v sinh tng hp, phng php ny c nhiu u im l cho cc dng L - amino acid l dng thng gp trong c th.
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Hnh 6.5. S sinh tng hp amino acid theo Lehninger
7. S trao i lipid
7.1. S phn gii lipid Lipid (ester phc tp ca glycerol v acid bo) v cc cht sp (ester phc tp ca acid bo v ru bc mt) c nhiu vi sinh vt dng lm ngun thc n v nng lng. So vi cc c cht khc th y l loi c cht c ng ha vi tc chm, qu trnh ny c thc hin bi nhiu nhm vi sinh vt khc nhau nh vi khun, x khun, vi nm
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Bc u tin ca qu trnh ng ha lipid l s phn gii thnh glycerol (hoc ru n nguyn t) v cc acid bo. Qu trnh ny c xc tc nh lipase ni bo hoc ngoi bo. Sau khi c phosphoryl ha, glycerol s c chuyn ha theo con ng EMP, cn cc acid bo i vo chu trnh - oxygen ha (Hnh 6.6). Nh vy, c qua mt vng oxygen ha han ton chui carbon ca phn t acid bo li mt i 2C. C tip tc nh vy th ton b chui carbon s c chuyn ha thnh acetyl - CoA, cht ny c chuyn ha nh chu trnh TCA hoc chu trnh acid glyoxygenlic. Mt s nm mc thuc cc chi nh Penicillium v Aspergillus c th oxygen ha acid bo mt cch khng trit v tch ly li trong mi trng metylketone to mi kh chu (gp trong du m b nhim mc).
Cu hi n tp ca chng 6
1. Cc hnh thc vn chuyn cc cht qua mng? 2. C ch ca qu trnh sinh nng lng ca h hp hiu kh v k kh? 3. Cc kiu h hp vi sinh vt?; Cc con ng phn gii hexose? 4. Cc kiu trao i cht vi sinh vt ? S phn gii cc hp cht protein ? 5. Phn bit ng ha v d ha. 6. Vai tr ca vi sinh vt trong qu trnh chuyn ha vt cht trong t nhin ?
* Ti liu c thm
* Ti liu tham kho

1. Nguyn Thnh t . 1999. Vi sinh vt hc, NXBGD, H Ni.
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2. Lehninger A.L., Nelson D.L., Cox M.M. 1993. Principles of Biochemistry. Second Edition. Worth Publishers. New York. 3. Prescot Harley Klein, 2002. Microbiology. W. C. Brown publisher, USA. 4. http://vietsciences.net v http://vietsciences.free.fr
*Gii thch thut ng

Trao i cht hay bin dng l nhng qu trnh sinh ho xy ra trong c th sinh vt vi mc ch sn sinh ngun nng lng nui sng t bo (qu trnh d ho) hoc tng hp nhng vt cht cu thnh nn t bo (qu trnh ng ho). D ha v ng ha - D ha (catabolism): cc qu trnh bin dng nng lng nhm to nng lng cho t bo - ng ha (anabolism): cc qu trnh bin dng vt cht thu nng lng nhm to ra cc n phn dng to cc hp cht i phn t sinh hc cho t bo ng ha v sinh tng hp cc ng n Khung carbon cng -c cung cp bi cc con ng d ha, c bit l ng phn v chu trnh TCA. - ng c cung cp t mi trng bn ngoi hoc c tng hp bn trong t bo, dng sinh tng hp vch t bo, polysaccharide, nucleic acid - S tng hp glucose t nguyn liu khng phi l carbohydrate c gi l gluconeogenesis: phosphoenol pyruvate t chu trnh TCA v hai cht trung gian l glucose 6phosphate v UDP-glucose ng ha v sinh tng hp cc amino acid - Hai mi amino acid c th c chia thnh 5 nhm da vo tin cht ca chng. Mi nhm c con ng sinh tng chung to thnh tin cht tng ng - Nitrogen c a vo hu ht cc amino acid bi cc phn ng chuyn amin dng nhm amine ca glutamate; glutamate c ti to bng NH3 v glutamate dehydrogenase ng ha v sinh tng hp cc nucleotide v axt bo - Sinh tng hp cc base nitric l qu trnh phc tp: purine c tng hp bng cch thm tng nguyn t vo khung ng-phosphate; vng pyrimidine c tng hp trc khi gn ng vo - Acid bo c tng hp bng cch thm dn cc phn t acetyl-CoA to thnh mch di vi s h tr ca acyl carrier protein (ACP)
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CHNG 7
H HP K KH I. KHI NIM CHUNG

Nhiu vi khun c th sinh trng c trong iu kin khng c oxygen, lc chng dng nitrate (hoc sulfate) lm cht nhn in t cui cng, v vy gi l h hp nitrate (hoc h hp sulfate). Kh nng chuyn in t cho nitrate v sulfate, gip cho vi khun thc hin tng i y qu trnh oxygen ha hon ton c cht, m khng cn c s tham gia ca oxygen phn t, nh vi sinh vt thu c nhiu nng lng hn so vi qu trnh ln men (Hnh 7.1)
Ngoi ra, c mt s t vi khun (Clostridium aceticum) li c th dng CO2 lm cht nhn hydro trong h hp k kh.
Hnh 7.1. S qu trnh h hp hiu kh, k kh v ln men
II. H HP NITRATE, AMMONIUM HA NITRITE V KH NITROGEN

H hp nitrate hay l kh d ho nitrate khc vi qu trnh kh ng ho nitrate ch, cc sn phm kh trong trng hp ny khng c t bo s dng tip, m thng c tit ra ngoi mi trng. Rt nhiu vi khun hiu kh khi sng trong iu kin k kh c kh nng dng nitrate lm cht nhn hydro cui cng, v d mt s loi thuc cc chi Bacillus, Aerobacter v E. coli kh nitrate thnh amoniac (qu trnh amon ho nitrate), trong khi mt s khc li kh nitrate gii phng nit phn t (N2) hoc (N2O), ngi ta cho rng qu trnh amon ho nitrate v phn nitrate ho c chung giai on u (Hnh 7.2)
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Hnh 7.2. S qu trnh h hp nitrate Vi khun nitrate ha thuc chi Nitrobacter dng nitrite lm ngun nng lng. C th biu th chui h hp ca vi khun nitrate nh sau.
Hnh 7.3. Chui h hp ca vi khun nitrate ha Nhiu loi vi sinh vt c kh nng dng nitrate lm ngun thc n nit, kh ng ho nitrate qua nitrit thnh amoniac, hp cht ny s tham gia tng hp trong t bo ca chng. NO 3 NO 2 x NH2OH NH 4 Khi ammonia ho nitrate ta c. 8[H] + H+ + NO 3 Cn khi kh nitrate ho. 10[H] + 2H+ + NO 3 N2 + 6H2O Khi h hp nitrate, c bit l khi phn nitrate ho, c cht hu c c oxygen ho hon ton n CO2 v H2O. Nng lng sinh ra vi nitrate l cht nhn hydrogen ch thp hn trong trng hp cht nhn l oxygen phn t khong 10%. ATP cng c to ra nh kt qu ca qu trnh phosphoryl ha trong chui h hp, v vy qu trnh phn nitrate ha m bo hot tnh sinh trng ca vi sinh vt mc cao. nhiu loi vi sinh vt c kh nng h hp nitrate nh E.coli th chui vn chuyn in t tng t nh trong trng hp hiu kh, ch c citocromo-oxygendase c thay bng nitratreductase. Giai on u ca qu trnh kh nitrate c xc tc nh nitratreductase. Nhiu vi khun phn nitrate ha c kh nng dng khng nhng nitrate m c nitrite lm cht nhn hydrogen, M. denitrificans cn kh c c N2O. Ty theo loi vi khun m sn phm ca NH 4 + OH- + 2H2O
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kh nitrate d ha v nitrite d ha l N2, N2O hay NO. Do vi khun phn nitrate ha c kh nng dng nitrate ch yu lm cht nhn in t, m khng c kh nng kh n thnh NH 4 , cho nn mun nui cy chng phi cn b sung thm ngun m vo mi trng (pepton hoc NH 4 ). Vi khun phn nitrate ha phn b rt rng trong t nhin, nhng loi thng gp nh. P. denitrificans, P. aeruginosa, P. fluorescens, Micrococcus denitrificans, Hydrogenomonas agilis... Vi khun phn nitrate ha l tc nhn sinh hc lm ngho nitrogen ca t (cn phn nitrate ha hc xy ra mnh khi t qu nhiu acid) qu trnh ny xy ra mnh khi t b k kh (t ngp nc, khng ti...)hoc khi dng phn m (nitrate) cng vi phn chung trn nhng rung la ngp nc, y phn nitrate dng bn cho la t hiu qu rt t, v nitrate c th mt ht rt nhanh, ch khong mi gi sau khi bn. Khi t thong kh qu trnh phn nitrate b c ch, v oxygen phn t c ch t bo vi khun tng hp enzym nitratreductase v nitritreductase.
III. H HP SULFATE
Phn ln vi sinh vt v thc vt c th dng sulfate lm ngun dinh dng lu hunh tng hp cchp cht ca c th cha S (acid amin cha S, enzym...). l qu trnh kh ng ha sulfate. Ch c mt s t vi sinh vt thuc 2 chi Desulfovibrio v Desulfotomaculum li dng sulfate lm cht nhn hydro cui cng trong h hp k kh. Qu trnh h hp sulfate cng tng t nh qu trnh h hp nitrate, nhng cc vi sinh vt thc hin qu trnh ny u thuc loi k kh bt buc, chng ch c th dng sulfate lm cht nhn hydro cui cng trong qu trnh oxygen ha c cht. H2S l sn phm ca qu trnh c sinh ra theo kh ng.
2 8[H] + SO 4
H2S + 2H2O + 2OH-
Vi khun thc hin qu trnh h hp sulfate trc khi b kh thnh H2S phi tri qua nhiu giai on trung gian m c nhng giai on cho n nay ngi ta cha bit c mt cch chc chn.
2 SO 4
APS
SO 2 3
S2-
(Adenosin-5-phosphosulfate) Nng lng sinh ra trong qu trnh h hp sulfate c vi sinh vt s dng ng ha cc hp cht hu c (acid hu c, amino acid) mt s vi khun kh sulfate ha c kh nng t dng cacbon, chng dng hydrogen phn t kh sulfate v s dng nng lng sinh ra trong qu trnh h hp sulfate ng ha CO2 trong khng kh. 4H2 + H2SO4 4CH3COCOOH + H2SO4 H2S + 2H2O + Q CH3COOH +4CO2 +H2S Vi khun kh sulfate ha c th phn gii pyruvate hnh thnh H2S. Vi khun kh sulfate ha thng gp nhng vng bn ly c kh H2S, tc l nhng ni c qu trnh phn gii k kh cc hp cht hu c. Trong 1ml nc bn cha 104 106 vi
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khun kh sulfate, cn trong bn c H2S s lng ca chng ln ti 107 t bo. Hng triu nm nay, chng tham gia tch cc vo qu trnh hnh thnh qung lu hunh, m du ha. Desulfotomaculum ruminis tham gia tch cc vo qu trnh kh sulfate v to thnh H2S trong nhng d dy c ca ng vt nhai li (tru, b...). Bng cch kh sulfate nhng vi sinh vt ca nhm ny tham gia tch cc vo chu trnh chuyn ha chuyn ha lu hunh trong t nhin. Mt khc, do sinh ra H2S c th lm c cht hng lot, H2S lm n mn cc b phn kim loi ca cc cng trnh chn su di t hoc di nc.
Thiobacillus denitrificans Thiobacillus thiooxidans Hnh 7.4. Mt s loi vi khun lu hunh
IV. H HP CARBONATE TO METHANE

Nhng vi sinh vt ny c kh nng hnh thnh methane t carbonate. Chng ly nng lng cho hot ng sng ca mnh t qu trnh oxygen ha k kh cc hp cht khong hay cc hp cht hu c n gin nh acid formic hay mt s acid bo bc cao. Nhng vi sinh vt sinh methane bao gm 4 nhm ph khc nhau v hnh thi. -Cc vi sinh vt sinh methane khng sinh bo t (Methanobacterium) -Cc vi sinh vt sinh methane sinh bo t (Methaneobacillus) -Cc vi sinh vt hnh cu sinh methane (Methanococcus) -Cc 8 cu khun sinh methane (Methanosarcina)
Methanosarcina acetivorans.
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Methanobacterium formocicum Methanospirillum sp.strain TM20-1 Hnh 7 .5. Hnh dng mt s loi vi khun methane Ngi ta thng gp nhng vi sinh vt sinh methane y nhng ao h, m v y bin, ni m iu kin k kh rt thun li cho chng pht trin. Methaneobacterium barkeri. C kh nng chuyn ho CO thnh CH4. Sn phm trung gian ca qu trnh chuyn ho l CO2 v H2.
Vi khun sinh methane c chng minh l loi vi sinh vt c kh nng sinh tng hp mnh vitamin B12. S gi thuyt v s hnh thnh CH4 t CO2 hoc Acetate vi khun sinh methane c th c trnh by nh sau.
Hnh 7.6. S gi thit sinh tng hp VTM B12 vi khun sinh methane
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V. H HP CARBONATE TO THNH ACETATE

Qu trnh h hp carbonate to thnh Acetate mi c pht hin gn y vi khun methane, kh sulfate.
Hnh 7.7. Hnh dng Vi khun Methanobacterium thermoautotrophicum Kt qu nghin cu vi khun Methanobacterium thermoautotrophicum cho thy nh sau.
Theo con ng ny, 1phn t CO2 b [H] c nng lc kh thnh CH3-X, 1phn t CO2 khc b CO dehydrogenase kh thnh CO. Qua qu trnh cacbocyl ha CH3X s sinh ra Acetyl-X, sau thnh Acetyl-CoA. Di s xc tc ca piruvate siterase, Acetyl-CoA s tip thu 3 phn t CO2 cacbocyl ha thnh pruvic acid. Pruvic acid thng qua cc con ng trao i cht quen thuc tng hp ra cc cht hu c cn tht cho t bo.
Cu hi n tp chng 7
1. So snh nhng im khc nhau gia h hp hiu kh v h hp k kh ? 2. Nhng im ging nhau v khc nhau gia h hp nitrate v h hp sulfate. 3. Vai tr ca vi khun methane trong thin nhin v trong i sng con ngi ? 4. in vo ch trng: a. Qu trnh phn nitrat ho l mt qu trnh c hi i vi nng nghip, bi v......... b. S phn nitrat ho xy ra mnh nhng ni oxy........................... 5. H hp k kh khc ln men k kh nhng c im no ? 6. Vai tr ca h hp sunfate i vi i sng con ngi ?
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* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc, NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty, 1997. Vi sinh vt hc, NXBGD H Ni.
* Ti liu tham kho

1. Nguyn Thnh t . 1999. Vi sinh vt hc, NXBGD, H Ni. 2. http://vietsciences.net v http://vietsciences.free.fr 3. Lehninger A.L., Nelson D.L., Cox M.M. 1993. Principles of Biochemistry. Second Edition. Worth Publishers. New York. 4. Prescot Harley Klein, 2002. Microbiology. W. C. Brown publisher, USA.
* Gii thch thut ng

Cc vi khun t dng thu nhn nng lng t phn ng oxygengen ha cc hp cht ha hc gi l vi khun ha dng (chemotroph). Nhng nhm s dng hp cht v c (nh nc, kh hydrogen, sulfua v ammonia) lm cht kh c gi l vi khun v c dng (lithotroph) H hp v ln men - H hp: in t t cht cho qua chui truyn in t c truyn n cht nhn in t cui cng ngoi mi trng nh O2, NO3+ ATP c to thnh theo c ch phosphoryl ha oxygen ha - Ln men: in t t mt cht hu c b oxygen ha c chuyn n sn phm ln men l cht hu c cn bng phn ng oxygen ha kh. ATP c to ra theo c ch phosphoryl ha c cht H hp k kh (anaerobic respiration) - H hp k kh: ha nng d dng (a s, cht cho in t l cht hu c), hay ha nng v c (dng H2 lm cht cho in t) dng chui truyn in t to ng lc proton, cht nhn in t sau cng khng phi l oxygen - ATP c to thnh t hn h hp hiu kh - Vi khun phn nitrate ha - Vi khun kh sulfate - Vi khun sinh methane - Vi khun sinh Acetate ng hnh
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CHNG 8
VI KHUN HA DNG V C HIU KH I. NITRATE HA

Cc mui ammonium c to thnh trong qu trnh ammonium ha protein, ur, kitin... c th c cy trc tip hp th, hoc s c chuyn thnh cc mui nitrate. Qu trnh ammonium ha mui ammonium thnh nitrate gi l qu trnh nitrate ha. Nitrate ha c thc hin do nhiu loi vi khun dinh dng v c c bit gi chung l vi khun nitrate ha.
1.Vi khun nitrate ha

Ngi u tin nghin cu vi khun nitrate ha v hot ng sinh l ca chng l nh khoa hc Nga Winogradsky. Nm 1889 ng chng minh vi khun nitrate ha cng nh vi khun lu hunh v vi khun st l nhng loi vi sinh vt dinh dng v c bng tng hp ha hc. ng cng khng nh qu trnh nitrate ha gm hai giai on. oxygen ha mui ammonium thnh nitrite v oxygen ha nitrite thnh nitrate. Hai giai on ny c thc hin do hai loi vi khun khc nhau. 1.1. Vi khun nitr oxygen ha mui ammonium thnh nitrite (bin NH3 NO 2 ) c ba chi chnh l Nitrosomonas, Nitrisocytis v Nitrosospira, Nitrosomonas c hnh cu hoc hnh bu dc ngn, Gram m, khng sinh bo t. Di ng c bng mt tin mao di. Nitrosospira l loi bin i nhiu hnh thi, nhng dng in hnh l trc khun gn ging du phy. 1.2. Vi khun oxygen ha nitrite thnh nitrate (bin NO 2 NO 3 )
Gm mt chi l Nitrobacter. Chng l trc khun nh khng c bo t, Gram m. Vi khun nitrate ha c th s dng c CO2 lm ngun C. Chng khng cn hp cht hu c c sn. Khng nhng th hp cht hu c cn lm c ch s pht trin ca chng. Trong phng th nghim mun nui cy vi khun nitrate phi dng mi trng ch cha nhng cht v c ( c mi trng c c th dng gel-silice thay cho thch). pH ca mi trng cng c nh hng n vi khun v qu trnh nitrate ha. pH thch hp nht l 8,5 i vi giai on th nht v 8,3- 9,3 i vi giai on th hai. Khi pH= 6 qu trnh nitrate ha khng tin hnh, do trong t chua s khng xy ra qu trnh ny. Nhit thch hp cho vi khun nitrate ha l 30- 370C.
2. C ch phn ng ca qu trnh nitrate ha.

Qu trnh oxygen ha mui ammonium n nitrate thc hin qua hai giai on. 1. Giai on oxygen ha mui ammonium thnh nitrite, c th biu th bng phng trnh tm tt nh sau.
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2NH3 + 3O2 dn dn.
2HNO2 + 2H2O + 158 kcal
Thc ra qu trnh c qua nhiu chng trung gian, NH3 mt H v nhn thm oxygen
Trong c hai giai on ca qu trnh nitrate ha u c gii phng nhiu nng lng. Vi khun nitrate ha s dng nng lng ny tin hnh phn ng kh CO2 thnh hp cht hu c sng song vi cc phn ng oxygen ha NH3 v HNO2. Qu trnh kh c th vit nh sau. CO2 + 4H + xkcal HCHO + H2 O
3. Cc yu t nh hng n qu trnh nitrate

- Vi khun nitrate ho l nhng vi khun hiu kh nn ch pht trin tt trong iu kin t thot thy tt. Do , trong iu kin t rung ngp nc NH4 c tch lu v khng chuyn ho thnh NO3 - Yu t quan trng c nh hng n hot ng ca vi khun nitrite ho v nitrate ho l pH ca t. pH thch hp cho hot ng sng ca vi khun ny thng trn 6, mc nitrate ho gim khi pH thp hn 6,0 rt thp pH=5, v ngng hn pH=4 hoc thp hn. Cc vi khun nitrate ha sng trong t chua, c mc pH ti u 6,5, cn vi khun ni t kim tnh c mc pH ti u 7,8. Ngoi nh hng hot ng ca vi khun, pH cn nh hng n mt s cc vi khun ny. Mt s vi khun nitrate ha tng dn theo mc tng pH t chua sang kim tnh.
Hnh 8.2. Vi khun Nitrosomonas (tri) v Nitrobacter (phi) Nitrosomonas europaea Nitrobacter winogradskyi
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NH3 + 1,5 O2 NO2- + H2O + H+
NO2- + 0,5 O2 NO3-
Do nh hng ny, vic bn vi cho t chua s lm gia tng tc nitrate ha cc m ammonium trong t. - Nhit cng c nh hng n hot ng sng ca nhm vi sinh vt nitrate ha ny. nhit thp hn 50C v cao hn 400C vi khun hot ng rt chm nn s chuyn bin m NH4 thnh m NO3 cng rt chm. Nhit ti u cho hot ng nhm vi khun ny nm trong khong 300C. iu ny gii thch c hin tng nitrate ha xy ra rt km vo ma ng v ma h, v xy ra rt mnh vo ma xun v ma thu vng n i.
4. ngha ca qu trnh nitrate ha

Qu trnh nitrate ha quan trng trong nng nghip v n bin mui ammonium thnh nitrate l ngun thc n N tt nht cho cy. Do kh nng dinh dng v c nn vi khun nitrate ha sng c nhng ch tng nh khng th c s sng nh granit, tng ni. N tham gia vo vic xm thc ni do tc dng n mn ca acid nitric c to thnh. Chng pht trin v lm ph hy mt phn tng gch v kin trc b tng.
II. OXYGEN HA CC HP CHT LU HUNH

Trong t nhin c mt s nhm vi sinh vt c th oxygen ha hp cht lu hunh v c, trong c nhiu loi thuc nhm t dng ha nng c loi l t dng quang nng v cng c loi d dng.
1. Vi khun t dng ha nng

Bng 8.1. Cht cho electron v pH thch hp ca mt s vi khun lu hunh Nhm vi khun lu hunh Thiobacillus thioparus T. denitrificans T.neapolitanus T.thiooxygendans T. ferrooxygendans Cht cho electron H2S, sulfite, S0, S2O 3 H2S, S0, S2O 2 3 S0, S2O 2 3 S0 S0, sulfite, Fe2+
2
pH thch hp sinh trng 6-8 68 68 2- 4 2- 4
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T. novellus T. intermedius Beggiatoa Thiothrix Thiomicrospira Thiosphaera Thermothrix Sunfolobus Acidianus
S2O 2 3 S2O 2 3 H2S, S2O 2 3 H2S S2O 2 ,H2S 3 H2S, S2O 2 , H2 3 H2S, S2O 2 , SO 3 3 H2S, S0 S0
6-8 3- 7 68 68 68 68 6,5 7,5 1- 5 1- 5
Vi khun lu hunh, trong nhiu loi thuc chi Thiobacillus, s dng mt loi hay
2 nhiu loi hp cht lu hunh (H2S, S, S2O 2 , S4O 6 ) lm ngun nng lng. Chng chuyn 3
cc hp cht lu hunh S nguyn t hoc cc mui sunfite cha SO 2 ), sau oxygen ha 3 thnh sulfate. Qu trnh sn sinh ATP v qu trnh sulfate ha c biu th nh sau.
Hnh 8.3. Qu trnh sn sinh ATP v qu trnh sulfate ha ca vi khun lu hunh. Ch thch. v l biu th v tr ca ATP sinh ra trn chui electron Cc bc oxygen ho hp cht lu hunh vi khun Thiobacillus cn c th biu th nh sau.
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Hnh 8.4. S minh ho cc bc oxygen ho cc hp cht S vi khun Thiobacillus Di tc dng ca Thiobacillus, lu hunh v mui ca n c th c chuyn ha nh sau.
Hnh 8.5. Vi khun Thiobacillus thioparus (tri - t bo; phi - khun lc)
2. Vi khun t dng quang nng

Nhng vi khun thuc b Pseudomonodales c 2 h Thiorodaceae v Chlorobacteriaceae. H Thiorodaceae gn ging nh vi khun mu tm. Chng c hnh que, hnh elip,
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hnh cong. a s c ln c 1-2x5-10 m, c loi di n 25-50 hoc 100 m. Chng sinh sn theo li trc phn, khng c nha bo, c tin mao v c th di ng. Chng thuc loi t dng quang nng, c th s dng CO2 lm ngun cacbon tng hp nn cht hu c ca c th, di tc ng ca nng lng nh sng mt tri. Phng trnh c th biu din nh sau. 2CO2 + H2S + 2H2O ...
As
(CH2O) + H2SO4
Trong qa trnh oxygen ha H2S, lu hunh c tch ly. Sau S c chuyn ha thnh SO4 v dn dn ra ngoi. Vi khun cng c th dng ng thi H2S nh cht cho H trong c qu trnh quang hp. Thuc h Thiorodaceae c vi khun Chromatium. l vi khun mu tm c nghin cu nhiu nht. T bo hnh bu dc di t 10 20 m. Rng khong 1 - 3 m, c tin mao c th di ng c. Trong t bo thng c ht S, a sng trong mi trng c H 2S trong bn. Thuc h Chlorobacteriaceae, c vi khun Chlorobium (c nghin cu nhiu nht). Kch thc khong 0,5-0,7x2,0-1,5 m. Vi khun khng hnh thnh nha bo, khng di ng, sinh sn theo phng thc phn ct v thnh chui, chung quanh c gic mc. N l vi khun k kh, c th oxygen ha H2S v cc hp cht S khc trong qu trnh quang hp. Nhiu vi khun thuc h ny c th oxygen ha H2S thnh S v trong c th khng c ht S, nhng S c tch ly ngoi c th. Phng thc thc hin qu trnh oxygen ha ca chng c th : CO2 + H2S
As
(CH2O) +2S + H2O
3. Vi khun d dng
Vi khun d dng chuyn ha S thng gp l Bacillus mesentericus, Bacillus asterosporus , Bacillus subtillis. Cc loi x khun v mt s nm, men c th oxygen ha S dng bt, nhng tc dng ca n rt yu.
III. OXYGEN HA ST
Trong t Fe2+ v Fe3+ lun lun chuyn ha ln nhau. iu kin oxygen ha kh ca t c nh hng ln n qu trnh ny. Khi thong kh, in th oxygen ha cao Fe2+ s c oxygen ha thnh Fe3+ . Fe2+ Fe3+ + e Hin nay ngi ta xc nh c cht rusticyanin, mt loi protein cha ng, trong chui h hp ca vi khun oxygen ho st. Chng cng vi cc enzym oxygen ho vi loi citocrom C v 1 loi citocrom a1 (c cha 2 hemoglobulin a) to thnh chui chuyn electron.
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Qu trnh chuyn electron khi vi khun thc hin vic oxygen ho st Fe2+ c th biu th nh sau.
Hnh7.6. S qu trnh chuyn electron khi vi khun thc hin vic oxygen ho st Fe2+ Trong iu kin ngp nc, iu kin in th oxygen ha thp th Fe3+ chuyn ha thnh Fe2+. S thay i ca qu trnh oxygen ha kh trong t khng ch l qu trnh ha hc n thun m cn chu tc ng ca sinh vt, trong vai tr ca vi sinh vt rt quan trng. Nu t ch ngp nc n thun th khng th no a in th oxygen ha kh xung thp. Nh vy, khi c qu trnh ngp nc c hot ng phan gii hp cht hu c th mi c th a in th oxygen ha kh xung thp chuyn Fe3+ thnh Fe2+. t trng la, trong thi gian ngp nc, Fe2+ tng rung chua, ln c th bin thnh d tan tng cng cho cy trng. Trong t c mt s vi sinh vt chuyn ha st thuc b Clamidobacteriales v b Eubacteriales. Thuc b Clamidobacteriales thng gp c Leptothrix v Crenothrix. Leptothrix. Vi khun a bo, cc t bo xp thnh chui di nm trong mt bao chung. Trn bao c oxyt st ngng t lm cho bao c mu vng hoc tm. N thng ni ln mt nc hoc bm trn que ci, hn , l cy. T bo trong bao c th t phn chia v t bo ny c th i ra ngoi, sinh tin mao v di ng. Leptothrix l loi k kh ty tin v t dng v c nhng ng thi cng c th dng cht hu c lm c cht cho h hp. Trong qu trnh t dng v c, n c c th ly nng lng t qu trnh oxygen ha Fe2+ thnh Fe3+ Crenothrix. Vi khun hnh bao, do nhiu t bo hnh que xp thnh chui. Trn bao c cc ht oxygent st. i lc bao c hnh si phn nhnh. N thng bm trn cc vt th v khng sinh bo t chyn ng nh Leptothrix. Gallionella. Vi khun thuc loi t dng, c th bin st Fe2+ thnh Fe3+ v ly nng lng. N khng th dng cht hu c. Trong qu trnh sng, vi khun ny thng sn sinh ra cht do v chnh cht do ny gip n dnh trn vt th nht nh. Trong cht do tit ra ca c th c nhng cht st ngng t, qu trnh ny c th tm tt nh sau. 2FeCO3 + O2 + 3H2O Fe2(OH)6 + 2CO2 +29,8 Kcal.
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Ngoi vi khun oxygen ha st Fe2+ thnh Fe3+ trong t nhin cng c mt s vi khun c th chuyn Fe3+ thnh Fe2+. T lu cc nh khoa hc pht hin mt s vi khun nh E. coli, Clostridium sporogenes trong iu kin k kh c th kh Fe(OH)3. Gn y c nhiu nghin cu cho thy cc vi khun Bacillus circulans, Aerobacter, Pseudomonas, mt s vi sinh vt ln men butyric cng c kh nng kh st. St b kh ch yu trong iu kin k kh. Qu trnh kh st c tin hnh l do trong qu trnh sng, cc loi vi khun trn lm cho mi trng chua, h thp in th oxygen ha kh, to iu kin st Fe3+ bin thnh Fe2+.
IV. OXYGEN HA HYDROGEN

S oxygen ha hydrogen phn t l mt qu trnh oxygen ha n gin nht. C th xem n l qu trnh trung gian gia s ln men v h hp hiu kh. Vi khun oxygen ha hydrogen cn gi l vi khun hydrogen, y l nhm vi khun t dng ha nng v c khng bt buc. Thng gp cc vi khun nhm Gram m nh. Acidovorax facilus, Alcaligenes eutrophus, A.ruhlandii, Aquaspirillum magnetotacticum, Pseudomonas carboxygendovorans, Hydrogenophaga flava, Seliberia carboxygendohydrogena, Paracoccus denitrifica, Aquifex pyrophilys... Cng c th thuc Gram dng nh. Bacillus schlegelii, Arthrobacter sp. Mycobacterium... Qu trnh oxygen ha hydrogen ca cc vi khun ny thc hin theo 2 giai on:
Ta thy trong qu trnh ny vi khun kh c CO2 thnh HCHO (cht hu c), v vy, nu mi trng thiu cacbon hu c th nhm vi khun ny s s dng CO2 lm ngun cacbon. iu ny cng lm cho lng cht hu c trong t c giu thm. Hin nay ngi ta xc nh ce nzyme xc tc cho qu trnh oxygen ha ny l hydrogenase. Enzym ny lm hot ha H2 v kh NAD+ H2 + NAD+ NAD.H(H+)
Theo chu trnh pentosephosphate ATP v NAD.H(H+) s s dng kh CO2 thnh cht hu c. Phng trnh tng qut l. 6H2 + O2 + CO2 [CH2O] + 5H2O cht hu c C 2 loi hydrogenase tham gia vo qu trnh ny: hydrogenase dng ht v hydrogenase d tan.
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b c Hnh 8.7. Mt s vi khun oxygen ha hydrogen a. Alcaligenes eutrophus; b. Aquaspirillum magnetotacticum c. Paracoccus pantotrophus
Cu hi n tp chng 8
1. Vai tr ca vi khun nitrate ha i vi cy trng ? 2. Nhng sai khc c bn gia cc nhm vi khun (Beggiatoa v Thiobacillus) oxygen ha lu hunh? 3. ngha ca vic nghin cu cc nhm vi khun oxygen ha Fe2+ v cc nhm vi khun oxygen ha H2 ?
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc, NXB i hc Hu.
* Ti liu tham kho

1. Nguyn Thnh t . 1999. Vi sinh vt hc, NXBGD, H Ni. 2. http://vietsciences.net v http://vietsciences.free.fr 3. Lehninger A.L., Nelson D.L., Cox M.M. 1993. Principles of Biochemistry. Second Edition. Worth Publishers. New York. 4. Prescot Harley Klein, 2002. Microbiology. W. C. Brown publisher, USA.
Vi sinh vt
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* Gii thch thut ng

Vi khun oxygen ha nitrite thnh nitrate (bin NO 2 NO 3 ) NO 2 )
Vi khun nitr oxygen ha mui ammonium thnh nitrite (bin NH3 Ha nng v c (chemolithotrophy)
- Thu nng lng bng cch oxxygen ha hp cht v c (hydrogen, sulfide, sulfur, ammonium, nitrite, ferrous ion) - c im: + T dng bng chu trnh Calvin + Dng s truyn in t ngc to ra lc kh (tr vi khun oxygen ha H2) khi t dng + Mt s va t dng, va d dng + Mt s va ha nng v c, va ha nng hu c hn dng (mixotroph) + Hu ht hiu kh, to ATP bng phosphoryl ha oxygen ha - Vi khun hydrogen (hydrogen bacteria) - Vi khun lu hunh (sulfur bacteria) - Vi khun oxygen ha st (iron-oxidizing bacteria) - Vi khun oxygen ha nitrogen
Vi sinh vt
181
CHNG 9
CC QU TRNH LN MEN I. CC QU TRNH LN MEN PH BIN VSV

Bng 9.1. Cc qu trnh ln men ph bin vi sinh vt
Kiu ln men Ln men ethanol Ln men lactic ng hnh Ln men lactic d hnh Acid propionic Acid hn hp Hexose Hexose Phn ng tng th 2 Ethanol + 2 CO2 2 Lactate + 2 H+ Vi sinh vt thc hin Nm men Zymomonas Streptococcus Mt s Lactobacillus Leuconostoc Mt s Lactobacillus Propionibacterium Clostridium propionicum Cc vi khun ng rut Escherichia, Salmonella, Klebsiella, Enterobacter Clostridium butyricum Clostridium acetobutylicum Clostridium kluyveri Clostridium aceticum Acetobacterium Methanothrix Methanosarcina
Acid butiric Butanol Caproat Homoacetic
Sinh methane
Hexose Lactate + Ethanol + CO2 + H+ Lactate Propionat + Acetate + CO2 Hexose Ethanol + 2,3-Butandiol + Xucxinat + Lactate + Acetate + Focmat + H2 + CO2 Hexose Butirat + Acetate + H2 + CO2 Hexose Butanol + Acetate + Aceton + Ethanol + H2 + CO2 Ethanol + Acetate + CO2 Caproat + Butirat + H2 Fructose 3 Acetate + 3 H+ 4 H2+ 2 CO2 + H+ Acetate + 2 H2O Acetate + H2O CH4 + HCO 3
Shigella,
II. TNH A DNG CA LN MEN

Bng 9.1 trnh by mt s kiu ln men chnh c phn loi da trn cc sn phm c to thnh. l cc qu trnh ln men ethanol, acid lactic, acid propionic, acid hn hp, acid butiric v acid homoacetic. C nhiu qu trnh ln men c phn loi da trn c cht c ln men thay v cc sn phm. Chng hn, nhiu vi khun k kh to thnh bo t (chi Clostridium) ln men cc amino acidvi s sn sinh Acetate, amoniac v H2. Cc loi Clostridium khc, nh C. acidiurici v C. purinolyticum ln men cc purin nh xantin hoc aenin vi s to thnh Acetate, focmat, CO2, v amoniac. Cn nhng bn k kh khc th ln men cc hp cht thm. Chng hn, vi khun Pelobacter acidigallici ln men hp cht thm phloroglucinol (1,3,5benzenetriol, C6H6O3) theo con ng sau . Phloroglucinol (C6H6O3) +3 H2O 3 Acetate- + 3H+, Go' = -142,5 kJ/phn ng Nhiu qu trnh ln men khng thng thng ch c thc hin bi mt nhm rt hn ch cc vi khun k kh, v trong mt s trng hp ch bi mt vi khun duy nht. Mt s v d c nu trn bng 9.2. Nhiu trong s cc vi khun ny c th c xem nh cc chuyn gia v trao i cht v chng c kh nng phn gii mt hoc nhiu c cht m cc vi
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khun khc khng phn gii c. Bng 9. 2. Mt s qu trnh ln men khng thng thng
Phn ng cn bng tng th Acetylen 2 C2H2 + 3 H2O Ethanol + Acetate + H+ Glycerol 4 Glycerol + 2 HCO 3 7 Acetate + 5 H+ + 4 H2O Resocxinol (mt 2 C6H4(OH)2 + 6 H2O 4 Acetate + hp cht thm) Butirat + 5 H+ Xinamat (mt 2C9H7O2 + 2 H2O C9H9O2 + Benseat hp cht thm) + Acetate + H+ Phlorogluxinol C6H6O3 + 3 H2O (mt hp cht 3 Acetate + 3 H+ thm) Putresxin 10 C4H12N2 + 26 H2O 6 Acetate + 7 Butirat + 20 NH4+ + 16 H2 + 13 H+ Xitrat Xitrat + 2 H2O Focmat + 2 Acetate + + HCO 3 + H Aconitat Aconitat + H++ 2 H2O 2 CO2 + 2 Acetate + H2 Glioxygenlat 4 Glioxygenlat +3 H+ + 3 H2O 6 CO2 + 5 H2 + Glicolat Xucxinat Xucxinat + H2O Propionat + HCO 3 Oxalat Oxalat + H2O Focmat + HCO Kiu ln men
3
Vi sinh vt thc hin Pelobacter acetylenicus Acetobacterium spp. Clostridium spp. Acetovibrio multivorans Pelobacter masiliensis Pelobacter acidigallici Cc vi khun k kh gram dng khng sinh bo t, cha phn loi Bacteroides sp. Acidaminococcus fermentans Vi khun gram m cha phn loi Propionigenium modestum Oxalobacter formigenes Malonomonas rubra Sporomusa malonica
Malonat
Malonat + H2O
Acetate + HCO 3
III. LN MEN KHNG C S PHOSPHORYL HA C CHT

Vi mt s c cht, nng lng sinh ra khng ghp i vi s tng hp ATP trc tip nh phosphorin ha c cht, tuy nhin cc hp cht ny vn tr gip sinh trng ln men ca mt c th no . Trong nhng trng hp ny s phn gii c cht c gn lin vi nhng bm ion c dng to nn mt graien proton hoc natri qua mng. V d v kiu ny c cc qu trnh ln men ca Propionigenium modestum hoc Oxalobacter formigenes, c hai u gn s ln men cc acid icacboxygenlic vi cc bm ion sinh nng lng lin kt vi mng. Propionigenium modestum thc hin phn ng sau y . Xucxinat2- + H2O Propionat- + HCO3- , Go' = -20,5 kJ/phn ng Phng trnh tng th ny thu khng nng lng t do ghp i vi s tng hp ATP trc tip nh phosphorin ha c cht, tuy nhin n li c dng lm phn ng thu nng lng duy nht cho sinh trng ca vi khun. S d nh vy l v s loi CO2 khi xucxinat (qua metilmalonil-CoA v decacboxygenlase lin kt vi mng bi Propionigenium modestum c ghp i vi s xut Na+ qua mng t bo cht. Mt ATPse chuyn v Na+ nm trn mng ca P. modestum s dng graien ny hot ha s tng hp ATP.
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Oxalobacter formigenes thc hin s ln men oxalat . Oxalat2- + H2O Focmat- + HCO3- , Go' = -26,7 kJ/phn ng pH trung tnh, oxalat tn ti di dng ion ha oxalat-, v s loi cacboxygenl thnh focmat- tiu th mt proton. Sau s xut focmat- khi t bo s to nn mt ng lc nh proton (proton motive force = PMF) c kh nng ghp i vi s tng hp ATP nh mt ATPase chuyn v proton nm trn mng. iu th v v c o trong trao i cht ca c Propionigenium modestum ln Oxalobacter formigenes nm ch s tng hp ATP xy ra khng cn c phosphoril ha c cht ln chui vn chuyn in t; tuy nhin, s to thnh ATP theo c ch ha thm vn xy ra da vo mt bm Na+/H+ kt hp vi s loi CO2 khi cc acid hu c. iu cn rt ra t nhng s ln men ny l . cc phn ng, d ch thu c di 31,8 kJ, con s cn thit to thnh mt ATP, hoc khng c kh nng ghp i vi s phosphorin ha c cht, cng khng th lp tc b coi l khng c kh nng tr gip sinh trng cho mt vi khun. Nu nh phn ng c th ghp i c vi mt graien ion th s sn sinh ATP vn c kh nng xy ra v do vy vn c sinh trng (Hnh 9.1).
Hnh 9.1. Ln men lin kt vi bm ion vi khun (nng lng thu c khng qua phosphorin ha c cht)
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IV. HIN TNG CNG DNG (syntrophy)

Trong vi sinh vt hc c nhiu v d v hin tng cng dng, mt hin tng m hai sinh vt khc nhau c th cng nhau phn gii mt cht no - v bo ton nng lng ca qu trnh phn gii n mc khng c th no trong hai c th mt mnh c th lm c. Trong a s trng hp, bn cht ca mt phn ng cng dng c lin quan ti kh dihydro (H2) sinh ra bi mt trong hai i tc v c tiu th bi i tc kia. Do vy, cho n nay, cng dng cng cn c gi l s chuyn H2 lin loi. K tiu th H2 c th l bt k mt c th no trong s cc vi khun kh sunfat, ln men homoAcetate, v vi khun sinh methane. Hy xem xt trng hp cng dng lin quan ti qu trnh ln men ethanol thnh Acetate v s sn sinh methane sau . Nh c th thy trn hnh 2, bn ln men ethanol thc hin mt phn ng khng thun li v mt nng lng t do ( Go' dng). Tuy nhin, H2 c sinh ra bi i tc ln men ethanol li l mt cht cho in t y gi tr i vi qu trnh hnh thnh methane do mt vi khun sinh methane thc hin. V khi cng hai phn ng li th phn ng tng th s l mt phn ng to nhit v c th tr gip sinh trng ca c hai i tc trong hn hp cng dng. Ln men ethanol 2CH3CH2OH + 2H2O Sinh methane 4H2+ CO2 Phn ng cng dng 2CH3CH2OH + CO2 CH4 + 2CH3COO- 111,3 kJ/phn ng CH4 + 2H2O - 130,7 kJ/phn ng 4H2 + 2CH3COO- +2H+ +19,4 kJ /phn ng
Mt v d khc ca hin tng cng dng l s oxygen ha butirat thnh Acetate cng vi H2 bi vi khun cng dng oxygen ha acid bo Syntrophomonas. Butirat- + 2H2O 2Acetate- + H+ + 2H2 , Go' = +48,2 kJ/phn ng S bin i nng lng t do ca phn ng ny rt khng thun li v trong ng ging thun khit, Syntrophomonas khng th mc c trn butirat. Song nu H2 va sinh ra trong phn ng c tiu th ngay bi mt c th i tc (nh mt vi khun sinh methane) th Syntrophomonas li sinh trng rt tt trong ng ging cy chung vi bn tiu th H2.
V. LN MEN RU NH NM MEN V VI KHUN

Ethanol l mt trong s cc sn phm ln men ph bin nht gp vi sinh vt. Bn thn thc vt v nhiu loi nm cng tch ly ethanol di cc iu kin k kh. Vi sinh vt sn sinh ethanol ch yu l nm men, c bit l cc chng thuc loi Saccharomyces cerevisiae. Ging nh a s nm, nm men l nhng c th h hp hiu kh, nhng khi vng mt khng kh chng s ln men hirat cacbon thnh ethanol v CO2. Nhiu vi khun k kh
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v hiu kh cng to thnh ethanol nh cc sn phm chnh hoc sn phm ph khi ln men cc hexose hoc pentose.
1. S to thnh ethanol nh nm men

S ln men glucose thnh ethanol v CO2 nh Saccharomyces cerevisiae din ra theo con ng Embden-Meyerhof. S chuyn ha piruvat thnh ethanol gm hai bc. Trong bc mt, piruvat c loi CO2 thnh acetaldehit nh piruvat-ecacboxygenlase vi s tham gia ca tiaminpirophosphate. Trong bc sau, acetaldehit b kh thnh ethanol nh alcoholedihydrogenase cha NADH.
Bng s chuyn H ny, dihydro tch ra do triosephosphate-dehydrogenase xc tc s c tiu th, nh vy trng thi oxygen ha-kh s c cn bng.
2. Cc dng phng trnh Neuberg

Cc phng php v pht hin ca Carl Neuberg v ln men ethanol khng ch c ngha lch s. ng khm ph ra rng, khng ch glucose m c piruvat cng c nm men ln men v acetaldehit xut hin nh mt sn phm trung gian c th b bt gi bi dihydrosulfit. Dihydrosulfit vn khng c i vi nm men. Nu b sung dihydrosulfit vo mt mi trng cha glucose ang c nm men ln men th acetaldehit s b kt ta do to thnh phc cht . CH3-CHO + NaHSO3 CH3- CHOH -SO3Na Lc glycerol s xut hin nh mt sn phm ln men mi, ng thi hiu sut ethanol v CO2 s gim xung. Ln men vi s c mt ca dihydrosulfit c s dng trong cng nghip sn xut glycerol. Nguyn l ca n l, acetaldehit b bt gi, v do vy khng th hot ng nh mt cht nhn in t c na. Lc ny, thay cho acetaldehit, dihydroxygenacetonphosphate c s dng lm cht nhn in t v b kh thnh glycerol-3-phosphate ri tip tc b loi phosphate tr thnh glycerol. Phng trnh phn ng din ra nh sau . Glucose + Dihydrosulfit Glycerol + Acetaldehit-sulfit + CO2 Phng trnh ln men ci bin ny c gi l dng ln men Neuberg 2.
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Khi b sung cc cht kim (NaHCO3, Na2HPO4) th glycerol cng c to thnh v acetaldehit b chuyn thnh ethanol v Acetate qua phn ng ha hai, v do vy khng cn hon thnh c chc nng ca mt cht nhn in t. Phng trnh ln men c tn l dng ln men Neuberg 3 v c vit nh sau . Glucose + 2H2O Ethanol + Acetate + 2Glycerol + 2CO2 Dng ln men thng thng c Neuberg gi l dng ln men 1.
3. Hiu ng Pasteur
Hiu ng Pasteur l s c ch ln men ethanol (hay qu trnh ng phn) bi oxygen (tc l bi h hp hiu kh). S chuyn sang h hp khng nhng lm gim s to thnh ethanol v CO2 m cn lm gim s hp th ng. C hai c ch chu trch nhim i vi hin tng ny. Mt mt, l s cnh tranh ADP v phosphate v c gia hai qu trnh phosphorin chui h hp v phosphorin ha c cht. S loi dihydro xy ra trong phn ng oxygen ha glyceraldehyt-3-phosphate cn ADP v phosphate v c (Pvc) . Glyceraldehyt-3-phosphate + NAD + Pvc 3-phosphoglicerate + NADH + ATP Do vy, s chuyn ha c cht (glucose) qua con ng EM ph thuc vo s c mt ca ADP v phosphate. Tuy nhin, trong iu kin hiu kh, s phosphorin ha chui h hp cng cnh tranh ADP v phosphate pht sinh ATP. Lc nng ni bo ca ADP v phosphate gim, v hu qu l, s chuyn ha glucose v do vy, s to thnh ethanol cng s gim. Khi dng initrophenol (DNP) lm bt hot phosphorin ha chui h hp, ADP v phosphate li tr nn c sn i vi phn ng loi dihydro ca glyceraldehyt-3-phosphate, bi vy ngi ta thy ngay trong iu kin thang kh, s tiu th glucose (cho ln men) vn tng. C ch th hai chu trch nhim i vi hiu ng Pasteur l c ch iu ho bng enzym d lp th. Hnh 3 tm tt nhng quan h iu ha ch cht trong hiu ng ny. Trong iu kin thong kh, s ln men ethanol ch to thnh mt vi ATP tnh trn mi phn t glucose. duy tr s cung cp nng lng, t bo phi s dng nhiu glucose. T l ca ATP/AMP trong trng hp ny rt thp. Tri li, khi oxygen xm nhp vo mi trng, ATP c to thm rt nhiu nh s qu trnh phosphorin ha chui h hp. Enzym cha kha d lp th iu ha s s dng glucose trong nm men ln men ethanol l phosphofructokinase. l enzym khng thun nghch u tin ca con ng EM. N xc tc cho s phosphorin ha fructose-6-phosphate thnh fructose-1,6-iphophat nh ATP v b km hm d lp th bi ATP. S km hm theo c ch lin kt ngc ny lm tng nng glucose-6-phosphate trong t bo do tnh thun nghch ca phn ng enzym trc . Glucose-6-phosphate km hm d lp th hexokinase, km hm c s hp thu glucose, do lm gim s tiu th ng ny. Trong khi ATP km hm d lp th phosphofructokinase th AMP li hot ha n. Bi vy mt t l ATP/AMP thp nh trong ln men li kch thch s hp thu v s trao i glucose.
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Hot tnh ca phosphofructokinase cng b km hm bi xitrat. Nh cht hiu ng m tnh th hai ny m tc dng km hm ca ATP c tng cng, tri li cc ion amn li kch thch enzym ny. S iu ha m t trn c th c chng minh d dng khi ngi ta o nng ni bo ca glucose, glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-diphosphate v cc triose-phosphate trc v sau khi chuyn t iu kin nui k kh sang iu kin nui hiu kh. Ngay sau khi c thng kh, nng glucose-6-phosphate v fructose-6-P ca t bo tng nhanh, cn nng ca fructose-1,6-diphosphate v cc triose-phosphate li gim mnh. y, phosphofructosekinase hot ng nh mt ci van c iu chnh bi cc aenilat cng nh cc cht trao i trung gian khc. Nu van b ng, cc cht trao i nm pha trn van s b ng, cn cc cht nm bn di van c tip tc chuyn ha v do vy, nng gim i. Hiu ng Pasteur cng gp vi khun (chng hn, E. coli) v m ng vt. Nguyn tc iu ha l ging nhau, ch c nhng khc bit v loi cht hiu ng. Chng hn, E. coli, khng phi AMP m ADP mi l cht hiu ng dng. Nhng c ch c bn ca trao i cht th rt ging nhau, cn cc c ch iu ha th thng c tnh c hiu loi v c hiu chng.
4. K thut sn xut ethanol nh nm men

Cc loi nm men dng trong sn xut ung cha cn l cc nm n bo mang nhiu c im phong ph v sinh trng v pht trin. Cc loi c quan tm trc ht trong sn xut cng nghip l Saccharomyces cerevisiae, Saccharomyces uvarum (carslbergensis), Schizosaccharomyces pombe v Kluyveromyces sp. Ngoi ra, nhiu bin chng bt ngun t cc nm men ny c to ra nh k thut di truyn.
a b c d Hnh 9.2. Mt s nm men dng trong sn xut ethanol: a. S. cerevisiae; b. Schizosaccharomyces pombe ; c. Kluyveromyces lactis ; d. Saccharomyces uvarum Hin nay ngi ta quan tm nhiu n vic to ra cc th t bin khuyt h hp t nm men nh vic s dng cc tc nhn ha hc nh cc thuc nhum hoc cc hp cht lm bin tnh protein. Cc bin chng ny cha ADN ti th b bin i theo hng c ch s to ra cc enzym cn thit cho s trao i cht hiu kh v do vy thch hp vi s tng hp ethanol.
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5. Hiu sut ln men
Hnh 9.3. Hiu ng Pasteur C ch iu ha nh enzym d lp th Phosphofructosekinase Nm men, di iu kin k kh chuyn ho glucose thnh ethanol trc ht bng con ng Embden-Meyerhof. Phn ng tht s bao gm s to thnh 2 mol ethanol, 2 mol CO2 v 2 mol ATP trn mi mol glucose c ln men. Do vy, v mt trng lng mi gam glucose v l thuyt c th to ra 0,51 g ethanol. Tuy nhin sn lng t c trong ln men thc t thng khng vt qu 90-95% con s l thuyt. S d nh vy l v mt phn cht dinh dng c s dng tng hp sinh khi mi v cho cc phn ng lin quan ti s duy tr t bo. Cng cn c cc phn ng ph xy ra (thng dn ti Glycerol v sucxinat) tiu th ti 4-5% c cht tng s. Nu loi tr c cc phn ng ny th sn lng ethanol c th tng thm c 2,7%. Mt nng nh oxygen cn phi c cung cp cho nm men ln men v y l thnh phn cn thit trong s sinh tng hp cc cht bo cha bo ha v cc lipit. Thng thng oxygen c gi trong dch ln men nng 0,05-0,1 mm Hg tnh theo p sut oxygen. Bt c nng no nm trn gii hn u s kch thch sinh trng, vi phm n s to thnh ethanol (hiu ng Pasteur).Cc nhu cu tng i ca cc cht dinh dng c dng trong s tng hp ethanol t l thun vi cc thnh phn chnh ca t bo nm men.
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Chng bao gm C, O, N, v H. Thp hn mt cht l P, S, K v Mg dng tng hp cc thnh phn th yu. Cc mui v c (nh Mn, Co, Cu, Zn) v cc nhn t sinh trng hu c (acid amin, acid nucleic, v cc vitamin) c yu cu di dng vt. Nhiu loi nguyn liu dng sn xut ethanol quy m ln (nh r ng ma hay actiso Jerusalem) ngoi hirat C cn cung cp ton b cc cht dinh dng cn thit cho sinh trng ca nm men. Trong mt s trng hp, ngi ta cng a thm vo mt s cht dinh dng b sung nh cc mui amn, phosphate K hay ngun cao ng r tin. Nm men rt mn cm vi s c ch bi ethanol. Nng 1-2% (w/v) lm chm sinh trng ca vi sinh vt v nng cn 10% (w/v) tc sinh trng ca nm men gn nh ngng hn. Bn cht ca hin tng km hm ny l rt phc tp. Vic thm cn vo pha log lm gim nhanh tc sinh trng ca nm men (c l do tc ng ln s tng hp protein), gim kh nng sng st ca t bo (qua s bin tnh khng thun nghch ca cc enzym), v mt mc thp hn nhiu, ethanol lm gim tc tng hp bn thn n. Cc dn liu v mc chu cn ca mt s chng nm men ph thuc rt nhiu vo thnh phn acid bo trong mng t bo cht cho thy rng thnh phn acid bo kch thch hoc km hm s tit ethanol khi bo tng. Cc kt qu khc cung chng minh c rng ethanol c to ra trong ln men (ethanol t sinh) c ch ln sinh trng ca t bo mnh hn l ethanol c a vo mt cch nhn to t bn ngoi.
6. S to thnh ethanol nh vi khun

Trong s vi khun, kiu ln men ethanol ph bin nm men (EM, piruvatecacboxygenlase) ch gp Sarcina ventriculi. y l loi vi khun chu kh yu, d dng phn lp c t t, nhng cng gp c trong dch tiu ha ca nhng ngi mc bnh d dy. Cc vi khun ny c coi l khng bnh thng v c kch thc rt ln (ng knh 4 m), to nn cc dng hp bao gi (cha trn 64 t bo mi hp , c lin kt vi nhau nh xenlulose), chu pH (sinh trng c pH t 0,9 n 9,8) v to thnh ni bo t.
T dch p ln men ca cy da di (Agave americana) vng Mexico ngi ta phn lp c mt loi vi khun hnh que, di ng, c lng roi hai cc, c kh nng to thnh ethanol. Vi khun ny c tn l Zymomonas mobilis, phn gii glucose theo con ng 2-keto-3deoxygen-6-phosphogluconat (KDPG) ct piruvat thnh acetaldehit v ioxygent cacbon. Acetaldehit c kh thnh ethanol. Ethanol, ioxygent cacbon v mt lng nh
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acid lactic l nhng sn phm ln men duy nht. Ngi ta rng trong ethanol mnh ch t da di, cc nguyn t cacbon s 2 v s 3 cng nh s 5 v s 6 thay th cc nguyn t cacbon s 1 v s 2 cng nh s 5 v s 6 trong ethanol ch t nm men. Trong qu trnh ln men ca mt s Enterobacteriaceae v Clostridium, ethanol xut hin nh mt sn phm ph. Tuy nhin, tin cht ca ethanol, acetaldehit, khng c gii phng trc tip t piruvat nh piruvatecacboxygenlase m c to thnh nh s kh acetilCoA. Cc vi khun ln men lactic d hnh (v d Leuconostoc mesenteroides) to ra alcohol theo mt con ng hon ton khc. Qua cc bc u tin ca con ng pentosephosphate (PP), glucose c phn gii thnh pentosephosphate. Phosphoketolase tn cng vo xilulose -5- phosphate . Xilulose-5-phosphate+Pvc Acetilphosphate + glyceraldehyt-3-phosphate
Acetilphosphate va to thnh nh acetaldehit-edihydrogenase v alcoholedihydrogenase s c kh thnh ethanol. Sn phm phn gii cn li l glyceraldehyt-3phosphate, s c kh thnh lactate qua piruvat.
Hnh 9.4. Vi khun Zymomonas mobilis v cy da di Agave americana
VI. LN MEN LACTIC V H LACTOBACTERIACEAE

Cc vi khun lactic c xp chung vo h Lactobacteriaceae. Mc d nhm ny khng c s ging nhau v hnh thi v, bao gm c vi khun hnh que di, hnh que ngn ln cc vi khun hnh cu, song v mt sinh l chng c chung cc c im kh c trng. Tt c cc i din u l cc vi khun gram dng, khng to thnh bo t (tr Sporolactobacillus) v khng di ng. thu nng lng chng hon ton ph thuc vo cc hirat cacbon v tit ra acid lactic (lactate). i lp vi cc Enterobacteriaceae l bn cng sn sinh acid lactic, chng thuc bn ln men bt buc. Chng khc cha cc hemin (xitocrom, catalase). Mc d vy, cc vi khun thuc h Lactobacteriaceae c kh nng sinh trng khi c mt oxygen khng kh, chng thuc bn k kh song ng thi cng l bn chu kh. Mt vi khun sinh trng hiu kh m
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khng cha catalase th gn nh chc chn l mt vi khun lactic (tr ngoi l Acetobacter peroxydans v Shigella dysenteriae).
1. Nhu cu v cc cht b sung v nhn t sinh trng

Mt c im na ca cc vi khun lactic l c nhu cu v cc cht b sung. Khng c i din no c th mc c trn mi trng v c thun khit cha glucose v cc mui amn. a s trong chng cn hng lot cc vitamin (lactoflavin, tiamin, acid pantotenic, acid nicotinic, acid folic, biotin), v cc acid amin, cc baz purin v pirimiin. V vy ngi ta thng nui chng trn cc mi trng dinh dng phc tp cha nhng s lng kh ln cao nm men, dch c chua, dch ng sa hoc thm ch c mu na. iu ngc nhin l mt s vi khun lactic (v c cc vi khun ln men khc) khi sinh trng trn mi trng dinh dng cha mu c th to thnh cc xitocrom v thm ch c th tin hnh qu trnh phosphorin ha chui h hp. V vi khun lactic thiu nng lc tng hp cc pocphirin nn khi cc pocphirin c b sung vo mi trng th mt s vi khun lactic c th to thnh cc sc t hem tng ng. Vi khun lactic c coi nh nhng k qu qut v trao i cht. C th rng do hu qu ca s chuyn ha sinh trng trong sa hoc trn nhng mi trng giu cht dinh dng v cht sinh trng, chng mt i kh nng tng hp nhiu cht trao i. Trong khi chng li c kh nng m a s cc vi khun khc khng c, l kh nng s dng lactose. Kh nng ny nhiu vi khun ng rut (v d Escherichia coli) cng c. Lactose khng c mt trong gii thc vt, n c cc ng vt c v to thnh v tit ra cng vi sa hoc c hp thu vo cng vi sa. Vic s dng c lactose cng c th c xem nh mt s thch nghi vi cc iu kin tn ti trong ng rut ca ng vt c v. Lactose l mt isaccarit, trc khi i vo cc con ng phn gii hexose n phi c phn ct .
Sau khi c phosphorin ha galactose c chuyn thnh glucosephosphate. Do c s sn sinh mt lng ln lactate nn mi trng dinh dng cn phi c trung ha. Thng ngi ta dng cacbonat canxi. Trn mt mi trng thch dinh dng c b sung cacbonat canxi c th nhn ra s to thnh acid qua cc vng trong sut bao quanh cc khun lc. Ni sng ch yu ca cc vi khun lactic l sa v nhng ni sn xut v ch bin sa, thc vt nguyn vn hoc ang t phn hy, cng nh ng rut v nim mc ca ngi v ng vt. Do to thnh mt lng ln lactate v do tnh chu chua m di cc iu kin mi trng thch hp cc vi khun lactic pht trin nhanh, v vy c th d dng phn lp cc vi khun lactic trn cc mi trng dinh dng chn lc.
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2. Ln men lactic ng hnh
Vi khun lactic ln men ng hnh to thnh ton b hoc gn nh ton b l lactate. Chng phn gii glucose qua con ng EM, cha cc enzym cn thit, k c alolase v chuyn dihydro tch ra trong qu trnh loi dihydro ca glyceraldehyt-3-phosphate sang piruvat .Vic xut hin D-, L+ hay DL- lactate l ph thuc vo tnh chuyn ha khng gian ca enzym vn chuyn lactate-edihydrogenase v vo s c mt ca mt lactate-racemase. Ch mt phn nh piruvat c loi cacboxygenl v chuyn thnh Acetate, ethanol, CO2 v acetoin. Vic to thnh cc sn phm ph ph thuc vo s c mt hay vng mt ca oxygen.
3. Ln men lactic d hnh
Hnh 9.5. Ln men lactic d hnh .1. Glucose-6-phosphat-dehydrogenase
2. Phosphogluconat-dedihydrogenase ;3. Epimerase ; 4. Phosphoketolase
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Vi khun ln men lactic d hnh thiu cc enzym ch yu ca con ng EM l alolase v triosephosphate-isomerase. on phn gii glucose u tin ch xy ra qua con ng pentosephosphate, tc l qua glucose-6-phosphate, 6-phosphogluconat v ribulose-5phosphate. Cht ny nh mt epimerase c chuyn thnh xilulose-5-phosphate v nh phosphoketolase c phn gii trong mt phn ng ph thuc taminpirophosphate lm xut hin glyceraldehyt-3-phosphate v acetilphosphate. Cc t bo Leuconostoc mesenteroides khng sinh trng v c ra sch ln men glucose thnh lactate, ethanol v CO2 theo mt t l gn nh tng ng. C6H12O6 CH3 - CHOH - COOH + CH3 - CH2 - OH + CO2 cc c th ny acetilphosphate b kh qua acetil-CoA v acetaldehit thnh ethanol. Cc vi khun ln men d hnh khc chuyn mt phn hoc ton b acetilphosphate thnh Acetate, trong lin kt phosphate giu nng lng c chuyn sang ADP v c s dng di dng ATP. Dihydro tha trong trng hp ny c truyn cho glucose lm xut hin manit. Glyceraldehyt-3-phosphate c chuyn qua piruvat thnh lactate. Ribose c L. mesenteroides ln men thnh lactate v Acetate. * Ln men lactic d hnh BifidobacteriumVi khun lactic d hnh Bifidobacterium bifidum c dng ch V hay ch Y (bifidus ting Hi Lp c ngha l b chia i, b phn ct), l thnh vin trong khu h ng rut ca tr s sinh, trc ht l cc tr nui bng sa m. Vi khun ny cn N-acetilglucosemin ch c trong sa ngi m khng c trong sa b. Cc i din thuc chi Bifidobacterium l nhng vi khun k kh nghim ngt, khng chu kh v cn mt kh quyn cha CO2 (10% CO2) sinh trng. Bifidobacterium chuyn ha glucose theo phng trnh . 2 C6H12O6 2 CH3 - CHOH - COOH + 3 CH3 - COOH Bifidobacterium ln men glucose qua con ng ph phosphoketolase, khng cha aldolase cng nh glucose-6-phosphat-dehydrogenase, song li cha cc phosphoketolase hot ng, phn gii fructose-6-phosphat v xilulose-6-phosphat thnh acetilphosphate v eritro-4-phosphate hoc glixeraldehyt-3-phosphate. Hexose c chuyn ha theo con ng sau y.
4. ng dng ca ln men lactic

Nu dung dch ng khng kh trng cha c cc ngun nit phc tp v cc cht sinh trng mt cch t nhin trong trong iu kin k kh hoc di dng lp cao th cc vi khun lactic s nhanh chng pht trin pha bn trong. Chng lm gim pH xung di 5 v qua c ch sinh trng ca cc vi khun k kh khc l bn khng c kh nng sinh trng chua ny. Trong cc mi trng lm giu ny nhm vi khun no pht trin c l tu thuc vo cc iu kin c bit. Nh c tnh bo qun v kh trng da trn nguyn tc acid ha m cc vi khun lactic c s dng, trong nng nghip, trong ni tr, trong ngnh sn xut v ch bin sa.
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4.1. Thc n chua. Vi khun lactic phn b trn cc nguyn liu thc vt gi mt vai tr to ln trong vic bo qun thc n gia sc. to iu kin cho ln men lactic xy ra, l cc loi cy ly c hoc c ... c nn cht trong cc hm , thm r ng nng cao t l C/N v to chua ban u bng cch thm acid formic hay acid v c. 4.2. Da L mt sn phm ca qu trnh ln men lactic. Bp ci c ct nh, trn v bp vi mui n 2-3%. Trong iu kin ym kh, ln men lactic s xy ra mt cch t nhin, lc u bi Leuconostoc - vi s to thnh CO2, v v sau c s tham gia ca Lactobacillus plantarum. 4.3. Cc sn phm sa. Sa c mt thnh phn dinh dng cao, cha nhiu nc v giu cc mui khong, protein (ch yu l cazein), m b, ng (c bit l lactose) v cc vitamin. trong bu v ca ng vt th n l v trng, nhng khi i ra khi nm v th sa b nhim bi khu h bnh thng ca ng vt, sau c b sung thm bi cc vi sinh vt khc t ngi vt sa, my hoc bnh cha. L mt mi trng gn nh hon ho nn sa rt mn cm vi s sinh trng ca vi sinh vt. Khi sa trong phng nhiu loi vi khun s ln men lactose to thnh acid v lm thay i tnh cht v kin trc ca sa. Qu trnh ny c th xy ra t nhin hoc c con ngi bt chc nh trong qn trnh sn xut fomage hoc iagua. Trong cc giai on sm ca ln men sa lactose b tn cng nhanh bi Streptococcus lactis v Lactobacillus spp. S tit acid lactic v s gim pH lm protein ca sa b ng li thnh cc vn (curd). S ng cng lm tch ring phn dch ng sa (whey) trn b mt. Cc vn cng c th c to nn nh tc dng ca vi sinh vt hoc nh mt enzym (rennin) ly t d dy b. Mt lt sa nng 1032g bao gm 902g nc v 130g cht kh trong cha 49g lactose; 39g cht bo (l nhng git m ng knh 1-8 m gm glixerit v cc cht tan trong lipit, nh colesterol v cc vitamin A, D, E, K; 32,7g protein m ch yu l cazein; 9g cht khong (gm phosphate, xitrat, KCl, NaCl v MgCl2). * Sa chua (sa b). Sa chua l sn phm ca s ln men sa lng bng s chua ngu nhin di tc ng ca nhiu vi khun sinh acid lactic khc nhau (nh Streptococcus lactis hay S. cremoris), hoc ca mt s lng t hn nhiu cc vi khun gy thm (nh Streptococcus diacetylacis, Betacoccus citrovorus) hoc bng vic cy vo sa un nng cc ging vi khun thun khit. Khi ln men din ra, lactose c chuyn ho thnh acid lactic, acid ny s ngng kt cazein pH 4-5. Sa ng, c, c v chua, ty theo vng c tn gi khc nhau. Sa chua c lm t sa y (cha t nht 3% m ca sa) thng c trn vi bt sa vt vng nhm lm tng hm lng cht rn ton phn v ci thin cu trc gel protein. mt s nc, sa chua cng c sn xut t sa cu, sa tru, sa tun lc hoc sa nga. Kem chua c sn xut bi mt qu trnh rt ging quy trnh dng sn xut sa chua ch tr vic kem c ph c s dng lm nguyn liu th.
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* Iagua. Iagua c lm t cc loi sa cha cc hm lng m khc nhau bng cch ln men nh cc ging c bit. N c mt ng c mn ging nh gel, mt chua r rt v hng v d chu, khc bit r rng vi loi sa chua bnh thng. Iagua c sn xut bng cch cy vo sa kh trng Pasteur ng nht khong 1,5-3% cc vi khun lactic a nhit (ging hn hp ca mt hoc nhiu chng nh Streptococcus thermophilus hay Lactobacillus bulgaricus) v sau gi 42-45oC trong khong 3 gi. qunh mn c hnh thnh trong thi gian ny. Sn phm cui cng c v chua ( chua 40-60oSH). Phn quan trng trong hng thm c bit ca iagua l cc hp cht cacbonyl vi ixetyl v acetaldehit chim u th. Mt loi igua c bit c tn l sa aciophilus c sn xut nh Lactobacillus acidophil. Vic thm ng v hoa qu vo iagua (iagua hoa qu) lm tng ng k sc tiu th sn phm ny. * Kefia v Kumis. Kefia v Kumis l cc loi ung cha ethanol giu CO2, nhiu bt. Khu h vi sinh vt bao gm nm men Torula (chu trch nhim i vi qu trnh ln men ethanol) v cc vi khun Streptococcus lactis v Lactobacillus caucaticus (chu trch nhim i vi ln men lactic). Cc ht kefia dng sn xut kefia cha cc cc sa ng cng vi cc vi khun kefia. Kumis c lm t sa nga hay sa d do cc ging kumis thun khit (Thermobacterrium bulgaricus v nm men Candida) . C hai ung lm t sa ny u l cc sn phm a phng ca vng Caucasus v cc vng tho nguyn Turkestan. Kefia cha acid lactic (0,5- 1%), mt s lng ethanol ng k (0,5-2%), CO2 v mt s sn phm phn gii cazein sinh ra t hot ng phn gii protein ca nm men. * Fomage. Fomage l sn phm thu c t sa ng bng cch loi b dch ng sa v bng cch lm chn cc vn khi c mt mt khu h vi sinh vt c bit. C nhiu loi fomage vi hnh dng v kch thc khc nhau. c xp thnh nhm da vo loi sa s dng (b, d, cu), s to thnh cc vn (dng acid, dng rennin hay dng c hai), kin trc hay c, v hm lng cht bo. Trong tng nhm, cc loi fomage ring bit c c trng bi hng thm. S sn xut fomage v c bn bao gm hai cng on . s to thnh cc vn v qu trnh lm chn. * S to thnh cc vn Hm lng cht bo ca sa c iu chnh ti mc mong mnn, v khi cn, hm lng protein cng c iu chnh. Cc cht ph gia bao gm mui canxi nhm ci thin s ngng kt protein v kin trc ca fomage, nitrat km hm khu h vi khun k kh hnh thnh bo t v cc sc t to mu. Sa th hoc sa kh trng c trn trong mt b nhit 15-59oC vi mt loi ging khi ng (vi khun lactic hay vi khun propionic, nm si, nh Penicillium camemberti, P. candidum, P. roqueforti , cc vi khun mang sc t mu vng hoc mu nh Bacterium linens, cng cc cu khun v nm men) v sa s ng li thnh mt khi bn lng mm, ccvn, sau ln men lactic, c ni tip bng s b sung rennin hoc mt t hp no m thng thng nht l t hp gia acid v rennin. Gel protein ny s c ct thnh cc hnh khi nh trong lc ang c un nng ri sau c khuy nh. Dch ng sa c loi b cn phn cc vn cha m th c chuyn sang qu trnh lm rn. Khi rn mong mun t c, cc vn c chuyn sang qu trnh lm chn.
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* Lm chn: Qu trnh lm chn ph thuc vo thnh phn khi fomage, c bit l hm lng nc, khu h vi sinh vt v cc iu kin bn ngoi nh nhit v m ca cc phng lm chn. S chn ca fomage mm xy ra t ngoi vo trong, v th trong giai on u s c mt phn v chn v mt phn li cha chn. S chn khng ng u xy ra l do c s chnh lch v pH. bn trong khi fomage, pH thp v c mt acid lactic. ngoi v, cc loi nm si c trng pht trin thch hp hn cc pH cao nng pH ln nh phn ng loi CO2 t cc amino acid. S chn fomage cng xy ra ng u trn ton b khi fomage. S to thnh v ch l kt qu ca s kh b mt, iu ny c th trnh c nh vic bao gi khi fomage trong cc mng plastic thch hp trc khi qu trnh lm chn bt u. Thi gian lm chn thay i ty trng hp, ko di t vi ngy i vi fomage mm ti vi thng thm ch vi nm i vi cc loi fomage cng. Sn lng trn 100 kg sa nc l 8 kg fomage cng v ti 12 kg fomage mm. Mi thnh phn ca fomage u chu s bin i v mt sinh ho hc cc mc khc nhau trong qu trnh lm chn. Lactose c phn gii thnh acid lactic nh ln men lactic ng hnh. Trong loi fomage chea chng hn, pH gim t 6,55 xung 5,15 tnh t lc b sung ging khi ng cho ti khi c thnh khun. Khi c mt vi khun propionic (nh trong trng hp ca fomage Emmental), acid lactic c chuyn ha tip tc thnh acid propionic, acid acetic, v CO2 theo phn ng . 3 CH3CHOHCOOH 2 CH3CH2COOH + CH3COOH + CO2 + H2O T l gia acid propionic v acid acetic b nh hng bi th oxygen ha kh ca fomage v t l ny s thp khi c mt mui nitrat chng hn. Phng thc v mc phn gii m ca sa ph thuc vo khu h vi sinh vt tham gia vo qu trnh lm chn fomage. S gii phng cc acid bo, c bit l cc acid nh hng n hng thm ca fomage ph thuc vo tnh c hiu ca cc lipase. Ngoi cc acid bo t do, 2-alkanon v 2-alkanol cng c to thnh nh cc sn phm ph ca s -oxygen ho cc acid bo. Nm si, c bit l Penicillium roqueorti, s dng -ketoaxyl-CoA eaxylase (tiodihydroase) v -ketoacid ecacboxylase cung cp cc hp cht in hnh cho hng thm ca cc fomage bn mm. S phn gii protein thnh cc amino acid xy ra qua cc peptit nh cc sn phm trung gian. Ty thuc vo loi sa m 20-40% cazein c chuyn ho thnh cc dn xut protein ha tan trong 5-15% l cc acid amin. Phm vi pH 3-6 l ti u i vi hot tnh proteinase ca Penicillium roqueorti. S phn gii protein b nh hng mnh bi hm lng mui cha trong fomage. Hm lng amino acid l 2,6-9% phn cht rn ca fomage v l phn quan trng trong hng thm ca fomage. Nhng li trong qu trnh lm chn c th to nn nhng peptit c v ng. Cc amino acid c chuyn ha tip tc. Trong giai on sm ca s lm chn, vi pH thp, chng b loi CO2 thnh cc amino. Trong giai on mun, vi pH cao, cc phn ng oxygen ho chim u th. S phn gii protein khng ch ng gp vo s to thnh hng thm, cn nh hng n kin trc ca fomage. Khi fomage mm b lm chn qu mc, s phn gii protein c th dn n vic dch ha gn nh ton b khi fomage.
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Cu hi n tp chng 9
1. Trong t bo, qu trnh ng phn din ra u? Phn ng ny khc nhau nh th no khi khng c mt oxy v khi c mt oxy? 2. Phn t no m u con ng ng phn ? Khi ng phn hon thnh, phn t no cn li. Bao nhiu ATP c to thnh mc phosphoril ha c cht ? Con s ATP c to thnh l bao nhiu, v v sao con s ny li khc vi tng s ATP c to thnh? Bao nhiu ATP s c to thnh b sung nu oxy c mt? 3. Hai c ch ti to NAD+ l nhng c ch no ? Chng khc nhau ch no? u l mt qu trnh hiu kh v u l mt qu trnh k kh? 4. Ln men khc h hp oxy ha ch no khi xt v phng din cc cht nhn in t v s sn sinh nng lng? 5. Hiu qu ca vic phn gii hiu kh hon ton mt phn t glucose vi khun l bao nhiu? So snh vi hiu qu ca mt mnh ng phn? 6. Hiu ng Pasteur c gp cc vi khun ln men lactic ng hnh khng ? V sao ?
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc, NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty. Vi sinh vt hc, 1997, NXBGD H Ni.
* Ti liu tham kho

1.. http://vietsciences.net v http://vietsciences.free.fr 2. Prescot Harley Klein, 2002. Microbiology. W. C. Brown publisher, USA.
* Gii thch thut ng :

Cc loi ung cha cn: Thng cc loi ung c cha cn c chia theo nng cn c bn trong: Kefia (kefir): c nng nhiu nht l 3% Bia: 1 12%, thng vo khong 5% Ethanol vang (vin): 7 14% thng vo khong 12% Ethanol mi: khong 15 75%, thng thng di 30% Ethanol mnh: thng vo khong 30 55%
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CHNG 10
VI KHUN QUANG HP V C NITROGEN PHN T I. VI SINH VT QUANG HP

1. Chu trnh carbon trong t nhin
Carbon tn ti di dng CO2, cc hp cht carbonate v hydrgencarbonate di cc i dng, cc ao h. S bin i CO2 thnh cc hp cht hu c nh qu trnh kh c thc hin nh hot ng quang hp ca cy xanh (phn ch yu) v ca vi to, ca vi khun quang hp. S bin i cc hp cht carbon hu c tr v trng thi v c l nh qu trnh oxygen ha, qua lm cho carbon trong t nhin c tun hon.
Hnh 10.1. Chu trnh n gin carbon trong t nhin Trong t nhin c nhiu nhm vi khun ha t dng v quang t dng. Ch yu l cc loi Thiobacillus thioparus, T. thioxidans. T. denitrificans, Beggiatoa minima chng l nhng VSV k kh, c th sng mi trng c pH thp, nhiu loi c th quang hp. S2O3- + 2H 2H2S + CO2 SO32- + H2S quang hp (CH2O) + 2S + H2O
Ta c th tm tt chu trnh carbon trong t nhin bao gm c k kh v hiu kh nh sau:
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Nh vy qu trnh s dng cc hp cht carbon chu nh hng rt nhiu khi c oxygen t do. Cc VSV hnh thnh cc sn phm khc nhau khi phn gii cc hp cht carbon trong iu kin hiu kh v k kh. Trong iu kin hiu kh cc sn phm oxygen ha c tch ly, trong khi cc sn phm kh c tch ly trong iu kin k kh, nh lm bin i hon ton c cht v m bo chu trnh carbon trong t nhin.
2. Cc vi khun quang hp (Phototrophic bacteria)

1. Vi khun lu hunh mu ta (Purple sulfur bacteria): 2. Vi khun khng lu hunh mu ta (Nonsulfure purple bacteria) 3. Vi khun lu hunh mu lc (Green sulfure bacteria) 4. Vi khun khng lu hunh mu lc (Green nonsulfur bacteria) 5. Vi khunlam(Ngnh Cyanobacteria) 2.1. Vi khun lu hunh mu ta (Purple sulfur bacteria) Thuc nhm ny l cc vi khun k kh bt buc, c kh nng quang t dng v c (photolithoautotroph), t bo c cha chlorophyll a hoc b , h thng quang hp cha cc mng hnh cu hay hnh phin (lamellar) gn vi mng sinh cht. dng lm ngun cho in t (electron donors) trong quang hp thng s dng H2, H2S hay S . C kh nng di ng vi tin mao mc cc, c loi chu mao, t l G+C l 45-70%. a - H Chromatiaceae:
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Chromatium sp.
Thiocapsa roseopersicina
Thiocystis
Thiospirillum
Lamprocystis
Thiopedia
Hnh 10.1: Mt s i din vi khun lu hunh mu ta thuc H Chromatiaceae
b- H Ectothiorhodospiraceae: 1. Chi Ectothiorhodospirace 2. Chi Halorhodospira
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(a) (b) (c) Hnh 10.2: Mt s i din vi khun lu hunh mu ta thuc H Ectothiorhodospiraceae (a) Ect. mobilis DSM 237; (b) Hlr. halophila BN 9621; (c) Halorhodospira halochloris 2.2. Vi khun khng lu hunh mu ta (Nonsulfure purple bacteria) Vi khun khng lu hunh mu ta l nhm vi khun quang d dng hu c (photoorganoheterotrophs) thng k kh bt buc, mt s loi l quang t dng v c khng bt buc (trong ti l ho d dng hu c- chemoorganoheterotrophs). T bo cha chlorophyl a hoc b, h thng quang hp cha cc mng hnh cu hay hnh phin (lamellar) gn vi mng sinh cht. dng lm ngun cho in t (electron donors) trong quang hp thng s dng cht hu c, i khi s dng hp cht lu hunh dng kh hoc H2. C kh nng di ng vi tin mao mc cc, hoc khng di ng, mt s loi c ti kh (gas vesicles), t l G+C l 61-72%.
Rhodospirillum
Rhodospirillum di KHV in t
Rhodopseudomonas
Rhodopseudomonas di KHV in t
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Rhodobacter
Rhodopila
Rhodocyclus purpureus
Rhomicrobium
Hnh 10.3. Mt s i din vi khun khng lu hunh mu ta 2.3. Vi khun lu hunh mu lc (Green sulfure bacteria) Thuc nhm ny l cc vi khun k kh bt buc, c kh nng quang t dng v c (photolithoautotroph), t bo c cha chlorophyll a cng vi b, c hoc e, cha caroten nhm 5, h thng quang hp lin quan n cc lc th (chlorosom) v c lp i vi mng sinh cht. dng lm ngun cho in t (electron donors) trong quang hp thng s dng H2, H2S hay S. Ht lu hunh tch lu bn ngoi t bo Khng c kh nng di ng, mt s loi c ti kh; t l G+C l 48-58%.
Chlorobium Pelodictyon Prosthecochloris Hnh 10.4. Mt s i din vi khun lu hunh mu lc 2.4. Vi khun khng lu hunh mu lc (Green nonsulfur bacteria) Thuc nhm ny l cc vi khun a bo, dng si,thng k kh khng bt buc ,thng l quang d dng (photoheterotroph), c loi quang t dng hoc ho d dng. T bo c cha chlorophyll a v c, trong iu kin k kh thy c chlorosom. dng lm ngun cho in t (electron donors) trong quang d dng l glucose, axit amin, axit hu c; trong
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quang t dng l H2, H2S. Di ng bng phng thc trn (gliding), t l G+C l 5355%. Chi in hnh l Chloroflexus, Chloronema
Chloronema
Chloroflexus
Hnh 10.5. Mt s i din vi khun khng lu hunh mu lc 2.5. Vi khun lam (Ngnh Cyanobacteria) Trc y thng nhm ln l To lam (Cyanophyta). Thc ra y l nhng c th nhn nguyn thu, khng lin quan g n to, ngoi kh nng quang hp hiu kh (quang t dng v c) v dng H2O lm cht cho in t trong qu trnh quang hp. Vi khun lam cha chlorophyll a v phycocyanin- phycobiliprotein. Mt s loi c sc t phycoerythrin. Chng phi hp vi sc t lc to nn mu nu. Mng lin kt vi phycobilisom. n bo hoc a bo dng si. Khng di ng hoc di ng bng cch trn (gliding), mt s loi c ti kh (gas vesicles). Nhiu loi c d t bo (heterocysts) v c kh nng c nh nit. Vi khun lam c mt khp mi ni, trong t, trn , trong sui nc nng, trong nc ngt v nc mn. Chng c nng lc chng chu cao hn so vi thc vt i vi cc iu kin bt li nh nhit cao, pH thp. Mt s loi c kh nng sng cng sinh vi cc c th khc nh Ru, Dng x, Tu...Nhiu loi cng sinh vi nm to ra a y (Lichen). Vi khun lam c th l sinh vt xut hin sm nht trn Tri t Vi khun lam c chia thnh 5 nhm (subsection) nh sau: a - Nhm I (B Chroococcales):
Chamaesiphon
Chroococcus
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Glooeothece
Gleocapsa
Prochloron
Hnh 10.6. Mt s i din vi khun lam thuc b Chroococcales b-Nhm II (B Pleurocapsales) Hnh que hoc hnh cu n bo. c th to dng kt khi (aggregate); phn ct nhiu ln to ra cc baeocytes; khng c d t bo.Ch c cc baeocytes l c di ng. T l G+C l 40-46% . Cc chi tiu biu l: Pleurocapsa , Dermocapsa, Chroococcidiopsis
Pleurocapsa
Dermocapsa
Chroococcidiopsis
Hnh 10.7. Mt s i din vi khun lam thuc b Pleurocapsales c-Nhm III (c tc gi gi l b Oscillatorriales): Dng si (filamentous) ; dng lng (trichome) khng phn nhnh ch c cc t bo dinh dng; phn i trn mt phng, c kiu t on (fragmentation); khng c d t bo; thng di ng. T l G+C l 34-67%. Cc chi tiu biu l: Lyngbya, Osscillatoria, Prochlorothrix, Spirulina , Pseudanabaena
Lyngbya
Oscillatoria
Prochlorothrix
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Spirulina
Pseudanabaena
Hnh 10.8. Mt s i din vi khun lam thuc b Oscillatorriales d-Nhm IV (c tc gi gi l b Nostocales) Dng si ; dng lng (trichome) khng phn nhnh c th cha cc t bo bit ho (specialized cell) ; phn i trn mt phng, c kiu t on to thnh on sinh sn (hormogonia) ; c t bo d hnh ; thng di ng c th sn sinh bo t mng dy (akinetes). T l G+C l 38-47%. Cc chi tiu biu l : Anabaena, Cylindrospermum, Aphanizomenon, Nostoc, Scytonema, Calothrix
Anabaena
Anabaena trong Bo hoa du
Cylindrospermum
Calothrix
Nostoc
Scytonema
Hnh 10.9. Mt s i din vi khun lam thuc b Nostocales
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e-Nhm V (B Stigonematales) :
Stigonema
Fischerella
Geitlerinema
Hnh 10.10. Mt s i din vi khun lam thuc b Stigonematales Lng (trichome) dng si, phn nhnh hoc do cc t bo nhiu hn mt chui to thnh ; phn i theo nhiu mt phng, hnh thnh on sinh sn (hormogonia); c t bo d hnh ; c th sn sinh bo t mng dy ( alkinetes), c hnh thi phc tp v bit ha (differentiation). T l G+C l 42-44%. Cc chi tiu biu l : Fischerella, Stigonema, Geitlerinema.
3. Trao i cht cc vi sinh vt quang dng

Tt c cc vi khun quang hp u cha sc t quang hp. Sc t quang hp vi khun c gi l bacteriochlorophyll. Chlorophyll v bacteriochlorophyll cn c gi l cht dip lc v cht khun lc. Cht dip lc, cht khun lc v huyt sc t c cu trc tng t nh nhau. l mt vng pocphiril do 4 nhn pirol lin kt vi nhau. Li ca cht dip lc v cht khun lc l Mg, cn li ca huyt sc t l Fe, cht dip lc a khc vi cht khun lc a,b,c,d,e gc R.. C th thy r hn s phn bit ca hai loi chlorophyll, bacteriochlorophyll bng cch quan st hnh sau y:
207
Bacteriochlorophyll a
Bacteriochlorophyll b
H.9.11: Cu trc ha hc ca mt s sc t quang hp Ngoi cc loi chlorophyll vi khun t dng quang nng cn c cha mt s cc sc t thuc loi carotenoit. Carotenoit vi khun khng ging vi carotenoit to hoc thc vt. Di y l vi v d:
H. 9.12. Cu trc ha hc cc loi carotenoit vi khun t dng quang nng c hai loi phosphoryl ha quang hp: phosphoryl ha quang hp tun hon v phosphoryl ha quang hp khng tun hon.
Ch thch: Bchl*: trng thi kch pht ca bacteriochlorophyll Ru-5-P: Ribulose-5-phosphate H2A: Cht v c cho hydrogen
H.9.13. Phosphoryl ha quang hp kiu tun hon
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Trong iu kin k kh mt s vi khun quang hp c th s dng nng lng ca nh sng thc hin phn ng phosphoryl ha sn sinh ra ATP. Electron t bacteriochlorophyll c tch ra di tc dng ca nh sng, sau tham gia vo chui h hp tun hon v quay tr li Bchl. Trn ng i sn sinh ra ATP. Vic sinh ra ATP c thc hin ring r vi vic sinh ra [H] c nng lc kh. [H] c nng lc kh c sinh ra t cc cht v c cho hydrogen (nh H2S...).Qu trnh quang hp ny khng sn sinh ra oxygen.
H.9.14. Phosphoryl ha quang hp kiu khng tun hon Ch thch: Chlb* v Chla* l trng thi kch pht. y l kiu phn ng phosphoryl ha quang hp gp vi khun lam v cng tng t nh to v cy xanh. y chui vn chuyn electron l khng tun hon v vy cn gi l dng chy electron khng tun hon. Quang hp xy ra trong iu kin c oxygen v c 2 h thng quang. H thng quang I c cha Chl a (cht dip lc a) v c th s dng c tia c bc sng di. H thng quang II c cha Chl.b v c th s dng c tia sng lam (bc sng ngn). Qu trnh quang hp ny c sn sinh oxygen v xy ra ng thi vic sinh ATP( h thng quang hp II) v vic sinh [H] c nng lc kh (trong NAD PH2) l H+ v e-, c sinh ra sau s quang gii nc. Sau quang gii nc sn sinh ra O2 +2H+ + 2e, electron s lin tip i qua chui electron (I v II), sau cng em electron n cho NADP+ sn sinh ra 2 phn ng phophoryl ha sinh ATP. h thng I c sinh NADPH2 v ATP, cn h thng II c sinh oxygen v ATP. Mt s vi khun a mn t dng quang nng c qu trnh quang hp kh dc bit. i din cho nhm ny l cc vi khun hay gp trn cc hi sn p mui (nh vi khun Halobacterium halobium, H.eutirubrum...) Mng t bo cht ca chng phn thnh 2 phn: phn mu v phn mu ta. Phn mu c cha cytocrom, flavoprotein.. ca chui h hp dng oxygen ha phosphoryl ha. Phn mu ta rt k l. Trn mng hin ln cc vt khoang (ng knh khong 0,5 m),
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phn b c lp vi nhau, chim ti mt na tng din tch ca mng t bo vi khun. Mng ny c kh nng thc hin mt qu trnh quang hp c o. C mt loi protein gi l bacteriorodopsin, rt ging vi rodopsin, loi protein c trong t bo hnh tr trn vng mc ca mt. Bacteriorodopsin chim ti 75% mng mu ta. Nhng vi khun ny c th sinh trng c trong ti hoc ngoi sng nu c mt oxygen, khi khng c mt oxygen chng ch sinh trng c ngoi sng. Chng c 2 con ng thu nhn nng lng: mt ng oxygen ha phosphoryl ha khi c oxygen v mt ng oxygen ha phosphoryl ha khi c chiu sng (quang hp). Tc to ra ATP l cao nht khi c chiu sng bng bc sng 550-600nm.
H.9.15. S minh ha 2 h thng quang Bng 9.1. Mt s tnh cht ca cc h thng quang hp VSV
II. C NH NITROGEN
Trong khng kh c rt nhiu nitrogen phn t, nhng tuyt i a s sinh vt khng s dng c ngun nitrogen ny. Ch c mt s VSV l c th hp th c nitrogen. Qua hot ng sng ca chng nitrogen s c chuyn thnh nitrogen hp cht (protein v cc sn phm thy phn protein). Hot ng ny c gi l s c nh nitrogen phn t.
210
Qu trnh c nh nitrogen c tc dng rt ln n i sng cc sinh vt trn tri t. Trn c bn trong nham thch khng cha hp cht nitrogen. Cho nn ngun nitrogen trong t hu nh hon ton do tc dng c nh nitrogen ca VSV m hnh thnh. Hng nm cy trng li ly i mt lng nitrogen ng k. Hp cht nitrogen c hnh thnh trong qu trnh c nh ny li chnh l ngun nitrogen ch yu b sung cho t. Vi sinh vt c nh nitrogen c ba nhm chnh: -Vi sinh vt cng sinh -Vi khun sng t do gm nhiu loi khc nhau -Vi khun lam
Nitrogen hu c
NH4 + NH4
+
C nh nitrogen N2 NH4 +
Phn nitrate ho
N2 O NO3 NO
NO2 -
Hnh 10.16. Chu trnh nitrogen trong t nhin
1. Vi khun c nh N cng sinh.

Phn ln cc cy thuc h u khng cn thc n hp cht, chng c th s dng c nitrogen khng kh. Nhng kh nng ny khng phi bn thn cy h u c m do nhng vi khun nt r cng sinh vi cy h u mang li. 1.1. Vi khun nt r Ngi u tin chng minh rng cc cy b u c th sinh trng trong t khng cha N l H. Hellrigel v H. Wilfarth (1886). Nm 1886 M.W.Beijerinck phn lp c vi khun t nt r mt s cy u. Ging (chi) Rhizobium (Franck, 1889) l loi vi khun n bo t, rt d nhm vi Pseudomonas, trc khun t do c kch thc 1,2(m x 0,5 chuyn ng nh nhung mao v tin mao cc, hiu kh. Khi hnh thnh nt sn th trc khun bin dng (c hin tng a hnh thi), n thay i hnh dng ty theo la tui v iu kin sng. Khi cn non chng l trc khun nh b, khng c bo t, di ng nh tin mao. Trong qu trnh pht trin, chng s chuyn thnh hnh cu v nhng dng phn nhnh hnh ch Y, ch V gi l gi khun th (Bacteroids).
211
Nm 1963, Manil chia cc vi khun nt sn thnh 3 nhm: -Rhizobium leguminosa (u H Lan), R. trifolii (c ba l), R. phaseoli (u c ve, u xanh). -Rhizobium lupini (mc tc, linh lng), gp nhm 1 v 2 thnh Rhizobium mc nhanh. - Rhizobium lupini (u a, lupin), R. Japonicum (u nnh) vi nhiu loi ph. Nhm ny hin nay c nh loi l Bradyrhizobium (gm tt c cc chng mc chm. Hin nay, ngi ta xp vi khun nt r vo mt chi chung l Rhizobium.Chng c th s dng nhiu hydratcarbon khc nhau lm ngun C. Khi sng ring l trong t hoc trong mi trng nui cy chng khng c kh nng c nh nitrogen. V h hp, chng l loi vi hiu kh. Rhizobium c nhiu loi v c tnh cht chuyn tnh, ngha l mi loi ch c th xm nhp v to thnh nt r trong mt s cy h u nht nh. Vi khun nt r khi gp cy h u thch hp s xm nhp vo t bo r, kch thch r to thnh nhng nt r. Nt r thng thng bng ht go, nt r to bng ht u nh, trong nt r cha y vi khun. Sau khi hnh thnh nt r gia vi khun v cy h u lm thnh mt th cng sinh c li cho c hai bn. Ngoi cc cy h u, mt s r cy khc cng c th cng sinh vi vi khun c nh N. Gn y ngi ta cn thy nt sn l mt vi loi cy nhit i. Trong nt sn cng cha vi khun c nh N gn ging r. 1.2. Phn bn nt r (nitragin) Phn bn vi sinh do Noble Hiltner sn xut u tin ti c nm 1896 v c t tn l nitragin: Sau pht trin sn xut ti mt s nc khc nh M (1896), Canada (1905), Nga (1907), Anh (1910) v Thy in (1914). Nitragin l loi phn c ch to bi vi khun Rhizobium do Beijerink phn lp nm 1888 v c Fred t tn vo nm 1889 dng bn cho cc loi cy thch hp ca h u. T cho n nay c nhiu cng trnh nghin cu nhm ng dng v m rng vic sn xut cc loi phn bn vi sinh c nh nit m thnh phn cn c phi hp thm mt s vi sinh vt c ch khc nh mt s x khun c nh nit sng t do Frankia spp, Azotobacter spp, cc vi khun c nh nitrogen sng t do Clostridium pasteurianum, Beijerinckia indica, cc x khun c kh nng gii cellulose, hoc mt s chng vi sinh vt c kh nng chuyn ha cc ngun d tr phospho v kali dng kh ho tan vi s lng ln c trong t mn, than bn, trong cc qung apatit, phosphorid v.v chuyn chng thnh dng d ho tan, cy trng c th hp th c. Trong cc loi vi sinh vt k trn, vi khun nt r c ngha ht sc to ln trong nng nghip. Cng c nh nitrogen cng sinh ln hn nhiu so vi cng c nh N khng cng sinh. Sau khi trng cc cy h u t giu thm nitrogen rt nhiu. Chnh v vy, t u th k XX ngi ta nghin cu s dng phn bn nt r vi mc ch l tip ging nhn to, lm cho t khng cha vi khun nt r thch ng s c vi khun thch ng, lm cho
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cy h u khng sinh nt r s c th sinh nt r v nhng cy c th sinh nt r th cng sinh nhiu hn, c nh N cng mnh m hn.
(b) (c) Hnh 10.17. Mt s vi khun c nh nitrogen a. Rhizobium sp.; b. Beijerinckia indica; c. Azospirillum brasilense Cch s dng phn bn vi khun nt r l em trn phn vi ht ging, ri kh, sau em gieo. Cng c th dng nt r tt, nghin nt trn vi ht ging. Khi s dng c th tng sn lng cy h u ln 15- 20%. Ngoi hai nhm vi khun c nh nitrogen sng cng sinh vi thc vt v sng t do trong t nh ni trn, cn mt s vi khun c kh nng c nh nitrogen sng trn b mt r v n su vo vo lp t chc b mt r ca mt s cy ha tho nh la, ng, ma, l mt loi vi khun c dng xon c pht hin t nm 1974 thuc chi Azospirillum. Ngoi cc nhm vi khun c nh nitrogen ni trn ra, cn c mt s loi vi to n bo cng c kh nng c nh nitrogen. V d nh vi khun lam sng t do v sng cng sinh trong bo hoa du. Cc loi ny cng ng gp khng nh vo qu trnh c nh N khng kh.
(a)
2. Vi sinh vt c nh nitrogen sng t do (khng cng sinh):

Bao gm mt s vi khun, x khun. Trong s ny quan trng nht l cc loi thuc chi Azotobacter v loi Clostridium Pasteurianum. 2.1. Azotobacter
(a) (b) Hnh 10.18. Hnh dng t bo Azotobacter sp. (a) di knh hin vi in t (b) tiu bn m Vi khun Azotobacter c khun lc nhy, li hoc tan, lc u khng mu, sau bin thnh mu nu ti, thm ch n mu en nhng khng lm nhum mu mi trng khun
213
lc. Ngoi ra mt s loi Azotobacter c dng nhn nheo, khun lc c mu vng lc, mu hng. V nhy ca vi khun Azotobacter cha khong 75% l cht hirit ca acid uronic v cha khong 0.023% N. Azotobacter l loi vi khun Gram m khng sinh bo t. Khi cn non t bo c dng que u trn ng ring l hay xp thnh tng i chng cht, cht t bo nhum mu ng u, c kh nng di ng nh tin mao, kch thc khong :2,0- 7,0 10- 2,5 m . Khi gi t bo Azotobacter mt kh nng di ng, kch thc thu nh li bin thnh dng hnh cu. Sinh cht xut hin nhiu ht ln nhn. l cc ht volutin, granulose, cc git m Lng ADN trong t bo Azotobacter thng thp hn so vi nhiu loi vi khun khc (0,70- 0,81%). 2.2. Clostridium pasteurianum v mt s VSV c nh N khc C.pasteurianum l trc khun k kh bt buc, c bo t, bo t nm gia lm t bo phnh ln nh hnh thoi. C. pasteurianum l mt loi vi khun butyric, chng c th s dng cc hydratcarbon thng thng v lm ln men suinh ra acid butyric, acetic, CO2 v H2. Chng c th s dng hp cht N v c v hu c lm ngun thc n N, khi nhng hp cht ny y th tc dng c nh N ca chng h thp hoc hon ton khng c. Tc dng c nh N ca C. pasteurianum thp hn Azotobacter. C mi khi tiu th 1g hydratcarbon th chng c nh c 2-3 mg N. Tuy nhin C.pasteurianum rt nhiu v phn b rng ri hn Azotobacter nn ngun N m chng c nh cho t l rt quan trng. Chng li pht trin c rung ngp nc, yu cu pH khng gt gao nn thch hp cho cc loi rung ca ta. Ngoi hai loi trn, trong cc VSV c nh N cn c mt s vi khun thuc chi Clostridium, Bacillus v Azotomonas. Tuy nhin, v nhng loi ny phn b khng nhiu v hiu lc c nh N cng thp nn tc dng thc t khng ln lm.
3. Vi khun lam (xem chng III )
Cu hi n tp chng 10
1.Hy nu tng qut chu trnh chuyn ha nitrogen trong thin nhin v vai tr ca vi sinh vt trong chu trnh ? 2.Cc nhm vi sinh vt c nh nitrogen ? ngha thc t ca vic nghin cu vi sinh vt c nh m. 3.Nu mt s thnh tu hin nay v nghin cu vi sinh vt c nh m. 4. Nitrogenase l g ? C ch hin bit ca qu trnh c nh nitrogen phn t. 5.Cc loi phn bn vi sinh vt ? c s khoa hc ca vic sn xut v s dng cc loi phn bn vi sinh vt? 6. Trnh by cc bc ch yu iu ch Azotobacterin v Nitragin. Trong iu kin hin nay Vit Nam c th sn xut 2 ch phm ny c khng? Ti sao?
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* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc, NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty. Vi sinh vt hc, 1997, NXBGD H Ni.
* Ti liu tham kho

1. Nguyn Thnh t , 1999. Vi sinh vt hc, NXBGD, H Ni. 2. http.//vietsciences.net v http.//vietsciences.free.fr
* Gii thch thut ng

- Vi khun quang hp khng thi oxygen (Anoxyphotobacteria): nhng vi khun thc hin qu trnh quang hp khng thi oxygen nh nhiu loi vi khun lu hunh. - Khun dip lc (Bacteriochlorophyll): sc t hp th nh sng tm thy nhiu loi vi khun lc v ta. - Quang phosphorin ha: (Photophosphory lation): qu trnh sn sinh ATP trong quang hp t ADP v phosphate v c (v cng vi NADPH+H+) cc t bo quang hp. - Chu trnh Calvin- Benson: con ng trao i cht ng ho CO2 v kh chng thnh glucid cy xanh v cc vi khun quang hp. - Quang tng hp (photosynthesis) - Cc phn ng sng: nng lng bc x ca nh sng c chuyn thnh nng lng ha hc - Cc phn ng ti: c nh (kh CO2) thnh vt liu t bo. Sc t v vai tr trong quang tng hp Chlorophyll: vng porphyrin cha Mg2+, mch carbon di k nc, d kt t vi mng - Cc chlorophyll khc nhau ch bi nhm th trn vng porphyrin, c bc sng nh sng hp thu cc i khc nhau - Hu ht t bo cha mt vi sc t khc nhau gip thu nng lng nh sng hiu qu hn - Mng quang tng hp, h quang, trung tm phn ng - Mng quang tng hp prokaryote c ngun gc t mng t bo cht; eukaryote v thc vt c hnh thnh trong dip lp th - Cc sc t ph tr Tng hp ATP trong quang tng hp
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- C ch quang phosphoryl ha: chui truyn in t, to c ng lc proton, tng hp ATP nh ATPase gn trong mng - Nng lng nh sng dng thc hin mt phn ng khng thun li v nhit ng hc l s oxygen ha sc t c th kh kh cao (khng c khuynh hng cho in t) Quang tng hp khng sinh oxygen - Vi khun quang nng lc v ta - Nng lng nh sng gip ly in t t bacteriochlorophyll truyn cho cc cht mang in t, cui cng quay tr v bacteriochlorophyll bi oxygen ha - in t c truyn trong mt vng kn: dng in t vng v phosphoryl ha vng - Nhiu vi khun l quang nng t dng khng sinh oxygen: lc kh CO2 (NADPH) c to ra bng s truyn in t ngc cn ATP - Mt s loi quang nng d dng khng sinh oxygen: ch thc hin phn ng sng thu nng lng, ngun C t hp cht hu c Quang tng hp sinh oxygen - To, vi khun lam - Hai loi chlorophyll khc nhau v hai h quang - Sau khi chlorophyll h quang II b oxygen ha do nng lng nh sng, phn t ny s b kh tr v trng thi bnh thng nh nhn in t t phn t nc; in t khng quay tr li phn t chlorophyll (dng in t khng vng v phosphoryl ha khng vng) - in t ca h quang II chuyn n chlorophyll h quang I b oxygen ha bi nh sng; sau in t ny c truyn vo chui truyn in t n ferredoxin (Fd) + Fd truyn in t cho NADP+ to NADPH + Fd truyn in t cho cyt bf h quang II v c truyn tr li chlorophyll ca trung tm phn ng II (dng in t vng v quang phosphoryl ha vng) Bin dng nitrogen - Nitrogen c cung cp t ngun hu c (nh amino acid) hoc v c - Vi sinh vt s dng m v c dng NH3 v NO3-, N2 - Khi nhiu NH3, glutamate dehydrogenase xc tc s gn NH3 vo cc hp cht hu c - Khi nng NH3 thp, glutamine synthetaseglutamate synthase chuyn NH3 v aketoglutarate thnh glutamate - C nh m N2 thnh NH3 c xc tc bi mt phc hp enzym l nitrogenase b bt hot bi O2
Chng 11: Di truyn hc vi sinh vt
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CHNG 11
DI TRUYN HC VI SINH VT I. KHI NIM CHUNG

Tnh di truyn l kh nng b m truyn cc c im (tnh trng) cho con ci ca mnh thng qua vic truyn vt cht di truyn l cc gene (hoc DNA). Bin d sinh hc l nhng bin i mi m c th sinh vt thu c do tc ng ca cc yu t mi trng v do qu trnh ti t hp di truyn. Vt liu di truyn (genetic material) ch cc i phn t ng vai tr lu gi v truyn thng tin di truyn qua cc th h t bo, hoc th h c th. cp t bo, vt liu di truyn l cc nhim sc th (chromosome). Cn cp phn t th l cc phn t DNA. B gene hay h gene, genome l tp hp cha ng ton b thng tin di truyn ca mt c th sinh vt c m ha trong DNA ( mt s virus c th l RNA). B gene bao gm nhng vng cha gene ln nhng on khng phin m. Thut ng genome c Hans Winkler gii thiu ln u tin nm 1920. Gene l on DNA mang mt chc nng nht nh trong qu trnh truyn thng tin di truyn. Thng thng, ngi ta ni n gene hm l gene cu trc. Gene cu trc l on DNA mang thng tin cn thit m ha mt chui polypeptide. Trong , cc polypeptide l thnh phn cu trc to nn cc protein. y l nhm phn t ng vai tr quan trng (nhng khng phi l hon ton) quy nh kiu hnh ca sinh vt. Intron l nhng an DNA bn Hnh 11.1. Hnh v minh ho v tr ca cc trong mt gene nhng khng tham gia vo exon v intron trong mt gene. vic m ho protein. Nhng vng cn li ca gene c gi l exon. Sau khi gene tin hnh phin m xong, nhng an intron s b loi b khi phn t mRNA bng c ch x l RNA (RNA splicing) trc khi cc mRNA c vn chuyn ra khi nhn t bo. Cc intron ch yu c mt trong cc t bo eukaryote. Lch s nghin cu Nm 1865, Mendel chng minh s tn ti ca cc nhn t di truyn (gene) trong cc t bo sinh sn v m u cho cc nghin cu i su vo th gii vi m ca s sng. Nm 1909, thut ng gene, kiu gene (geneotype), kiu Hnh (phenotype) c t ra bi nh sinh hc an Mch Wilhelm Johannsen (1857-1927).
217
Nm 1868, nh khoa hc Thy S Friedrich Miescher (1844-1895) pht hin ra trong nhn t bo m c 1 loi axit cha pht pho -nuclein m ngy nay gi l a.nucleic. Nm 1891 Kossel, Albrecht (1853-1927)(c) tm ra DNA.
Gr. Mendel
W. Johannsen
Friedrich Miescher Frederick Griffith
J.D.Watson
Francis Crick Erwin Chargaff Barbara McClintock Hnh 11.2. Mt s nh di truyn hc xut sc
Nm 1920, Hans Winkler (1877-1945) a ra khi nim genome (tp hp cha ng ton b thng tin di truyn ca mt c th sinh vt) 1928, Frederick Griffith (1871 - 1941), ngi u tin pht hin ra nhn t di truyn vi khun l DNA. Nm 1941, G.Beadle v E.Tatum nu ra thuyt 1 gene- 1 enzym, chng minh gene kim tra cc phn ng sinh ha Nm 1944, ba nh khoa hc Oswald T. Avery (1877-1955) Colin MacLeod (19091972) Maclyn McCarty (1911-) pht hin ln 2 nhn t di truyn l DNA Nm 1951, b Barbara McClintock (1902- 1992) nh di truyn hc ngi M pht hin cc gene di chuyn dc theo nhim sc th gene nhy. B nhn gii Nobel nm 1983 tui 80. Nm 1953, Watson-Crick tm ra m Hnh cu trc DNA v c coi l pht minh ln nht th k 20, to ra bc ngot mi cho di truyn hc v sinh hc. Nm 1957, Francis Crick xut quy tc trung tm ca sinh hc phn t. Nm 1961, Mt m di truyn th c pht hin bi 2 nh sinh ha M S.Ochoa v
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M. W.Nirenberg (Nobel 1968). 1973, ln u tin Cohen v Boyer s dng plasmid to dng DNA E. coli. Nm 1970 n nm 1973, Har Gobind Khorana tng hp c mt gene nhn to c th hot ng bnh thng trong t bo sng. Nm 1970, Temin Howard Martin (1934-1994) pht hin enzym phin m ngc (revertranscriptase) mt loi enzym quan trng cho k thut di truyn hc.
Hnh 11.3. S minh ha hc thuyt trung tm (central dogma) ca sinh hc phn t 1973- Ames trin khai phng php vi sinh vt hc khm ph ra cc yu t gy t bin (mutagenes); Cohen, Boyer, Chang v Helling s dng vect plasmid tch dng gene vi khun. 1975- Kohler v Milstein pht trin k thut sn xut cc khng th n dng ( monoclonal antibodies). 1977- Woese v Fox tha nhn Vi khun c (Archaea) l mt nhm vi sinh vt ring bit; Walter Gilbert v Frederick Sanger trin khai k thut gii trnh t DNA (DNA sequencing) 1983-1984- Gallo v Montagnier phn lp v nh loi virus gy suy gim min dch ngi 1995- Gii trnh t h gene ca vi khun Haemophilus influenzae. 1996- Gii trnh t h gene ca vi khun Methanococcus jannaschii; Gii trnh t h gene nm men S. cerevisiae gm 6 000 gene. 1997-Pht hin ra loi vi khun ln nht Thiomargarita namibiensis; Gii trnh t h gene vi khun Escherichia coli gm 3 200 gene. 2000- Pht hin ra vi khun t Vibrio cholerae c 2 nhim sc th ring bit. 2005- Gii Nobel c trao cho Barry J. Marshall v J. Robin Warren pht hin ra vi khun Helicobacter pylori l tc nhn gy bnh d dy v hnh t trng. Gii Nobel Y hc 2006 dnh cho hai nh khoa hc M Andrew Z. Fire v Craig C. Mello ca M c cng khm ph cch v hiu ha nh hng ca cc gene c bit.
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II. C S VT CHT DI TRUYN VI SINH VT

1. Cu trc genom ca sinh vt nhn s
DNA trong genom nhn s c tm thy di hai dng cu trc: nhim sc th v cc plasmid. 1.1. Nhim sc th nhn s Cc t bo nhn s c t 1 n 2 nhim sc th.
Mt nhim sc th vi khun in hnh cha 1 si DNA dng vng nm trong mt vng ca t bo c gi l th nhn. Th nhn khng c mng bao quanh.DNA nhim sc th c gp np thnh nhng ci mc di t 50.000 n 100.000bp c gi c nh bng cc phn t protein v RNA. Ton b NST li c gp np ln na v c cun li ging nh mt cun ch to thnh mt khi rn chc. Gn y, cc nh khoa hc cho bit mt s loi vi khun cha 2 NST. Agrobacterium tumefaciens, mt vi khun c dng chuyn gen vo thc vt cha 2NST, mt dng vng v mt dng mch thng. Mt v d khc v sinh vt nhn s cha 2NST l Vibrio cholerae, vi khun gy bnh t.
Hnh 11.4: Hnh dng vi khun Vibrio cholerae 1.2. Plasmid v episom * Cc plasmid Ngoi NST, nhiu t bo vi khun cn cha mt oc nhiu plasmid, l cc phn t DNA mch i nh dng vng (hnh 11.5) c th sao chp c lp ngoi NST. Plasmid c dng vng v thng c di bng 1%-5% di ca mt NST vi khun, dao ng v kch thc t vi nghn bp n vi triu pb. Chng c th hin din ch mt bn sao, i vi plasmid ln, cho ti vi trm bn sao trong cng mt t bo. Mi plasmid mang cc thng tin di truyn cn thit cho s sao chp bn thn n, v thng cho 2 tnh trng ca t bo hoc hn. Thng thng, cc gen nm trn cc plasmid khng cn thit cho s trao i cht bn thng, cho sinh trng hay cho sinh sn ca t bo, song cc gen ca plasmid c th to ra nhiu u im cho t bo mang chng.
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Hnh 11.5. S minh ha mt t bo vi khun vi plasmid bn trong (1) DNA nhim sc th. (2) Plasmids * Episomes l nhng plasmid c kh nng gn xen vo DNA nhim sc th ca sinh vt ch (Hnh 11.6). Nh kh nng ny, chng c th tn ti trong mt thi gian di, c sao chp cng lc vi DNA nhim sc th khi t bo phn chia, v tr thnh mt phn trong b my di truyn ca t bo. Thut ng ny khng cn c dng cho plasmid, v gi y ngi ta bit trnh t tng ng (homology) vi nhim sc th trn plasmid, nh transposon, bin plasmid thnh episome (gip plasmid gn xen vo nhim sc th).
Hnh 11.6. So snh plasmid khng gn xen (trn) v episomes (di). 1 DNA nhim sc th; 2. Plasmids; 3. Phn bo; 4. DNA nhim sc th vi plasmid gn xen vo * Cc loi plasmid 1. Phn loi plasmid da vo chc nng : C 5 nhm chnh: 2. Plasmid gii tnh (Fertility-(F) plasmid), mang cc tra gene, c kh nng tip hp. 3. Plasmid mang tnh khng (Resistance-(R) plasmid), mang cc gene c kh nng khng li cc thuc khng sinh hay cc cht c. c bit di thut ng Rfactor trc khi pht hin ra bn cht ca n l plasmid.
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4. Col-plasmid, cha gene m ha cho s tng hp colchicine, mt protein c th git cht cc vi khun khc. 5. Plasmid phn hy, gip phn hy cc cht l nh toluene hay salicylic acid. 6. Plasmid mang c tnh, lm cho sinh vt tr thnh sinh vt gy bnh. 7. Mt plasmid c th thuc mt hoc nhiu nhm chc nng k trn.
Hnh 11.7. S v tip hp vi khun 1 DNA nhim sc th. 2 Plasmids. 3 Pilus.
Hnh 11.8. Bn enzym hn ch ca plasmid pBR322, l mt plasmid c s dng

ph bin trong k thut DNA ti t hp. Plasmid ci bin nhn to ny mang hai gene khng thuc (tetR khng tetracycline v ampR khng ampicillin) v mt lot v tr ct ca enzym hn ch trong mi enzym ny ch c mt v tr phn ct, nh vy plasmid vector ny p dng c vi nhiu DNA khc nhau.
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2. Vt cht di truyn virus

H gene ca virus rt c bit (bng 2.3), mi loi virus ch cha mt loi acid nucleic, hoc l DNA hoc l RNA, chui n hay chui kp, dng si thng hay dng vng. Virus b nht khong 4 gene v ln nht khong vi trm gene. Mt s virus ngoi v protein cn c thm vi lp mng bao khc gm (protein + phospholipid) bt ngun t mng sinh cht t bo ch, hoc c (protein + glycoprotein) t virus HIV, virus SARS, H5N1 l mt v d. Bng 10.1. Dng acid nucleic v kch thc h gene mt s virus Virus Phage MS2, f2, R17 Virus TMV Cc Reovirus Phage X174 Vt ch E. coli Thuc l Th E. coli Th Cn trng Ngi E. coli E. coli Dng acid nucleic RNA si n-thng RNA si kp-thng DNA si n-vng DNA si kp-vng DNA si kp-thng Kch thc h gene ( s cp base) 3.569 ~ 6.000 ~ 50.000 5.375 5.243 (100-150)x103 ~ 36.000 48.502 ~ 200.000
Virus SV40 Baculovirus Adenovirus Ad5 Phage (Lambda)
Phage T2, T4
Ghi ch: Cc virus gy bnh ngi v ng vt c h gene ch yu l DNA si kp-thng,

nh: cc virus thuc h Adenoviridae gy bnh ng h hp, h Herpesviridae gy mn rp herpes ngi, bnh u g, h Poxviridae gy u ma, u b ... Hoc l RNA si n-thng, nh cc virus thuc h Retroviridae gy ung th, AIDS, bch cu ..., h Paramyxoviridae gy si, quai b, bnh g toi, h Rhabdoviridae vi bnh di, h Orthomyxoviridae vi bnh cm, h Picornaviridae vi cc bnh vim ty xm, vim gan A do virus, bnh l mm long mng tru b...
1.2. Phng thc sinh sn v vng i ca virus y chng ta s tm hiu nhng nt chung nht trong chu trnh sng-sinh sn ca virus qua i din l cc phage k sinh E. coli. Thng thng c chia lm 5 giai on: Hp th Xm nhp Sinh tng hp Lp rp Gii phng, hoc 3 giai on nh sau: (1) Phage bm bng phn ui trn b mt t bo vi khun v tit enzym tiu hy vch t bo; (2) Phage dng phn ui tim li acid nucleic (DNA) vo trong t bo ch; (3) Trong t bo ch, ty loi DNA cc phage khc nhau m c th din ra mt trong hai trng thi: gy tan hoc tim tan. + i vi cc phage c nh T2 v T4, lc ny DNA ca chng s sao chp nhiu ln v tng hp cc protein cn cho to v v ui, sau chng lp rp vi nhau to ra cc
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phage th h con. Sau , di tc dng ca lysozyme lm tan t bo ch v phng thch chng 100-200 phage con.
Hnh 11.10. Chu trnh hot ng gy tan (lytic cycle) v tim tan (lysogenic cycle) ca phage E. coli. + i vi virus thuc nhm retrovirus (virus gy ung th, gy bnh AIDS, gy bnh bch cu t bo T...) c chu trnh sng in hnh (hnh 2.11).
Hnh 11.11. Chu trnh sng ca virus nhm retrovirus y cn lu : + Ti t hp: Khi c nhiu virus nhim ng thi t bo ch th gia chng xy ra s trao i gene, to ra cc th ti t hp. Nh vy c th lp bn di truyn virus. Dng ti t hp im c hiu gn DNA phage vo nhim sc th vi khun ch v tch ra ( khi b cm ng tia UV) c xc tc bi cp enzym tng ng l integrase v excisionase. Li dng c im ny, ngi ta s dng phage lm vector trong k thut gene. + Sao chp:
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Do vt liu di truyn ca cc virus rt a dng, nn s sao chp/sao chp ca chng cng khc nhau, theo 3 con ng: DNA DNA; RNA RNA; v RNA cDNA kp RNA. Trong , cc retrovirus c h gene RNA gy ung th, bnh AIDS (HIV), bnh bch cu t bo T (HTLV-I)...u c cha enzym phin m ngc (reverse transcriptase) tng hp cDNA si n trn khun RNA ca n, ri sau l cDNA si kp c th xen vo nhim sc th vt ch trng thi provirus n nh. Li dng enzym ny tng hp v to dng cDNA. + Phin m:
Hnh 11.12. S minh ha qu trnh phin m mt s virus
3. Cu trc genom ca sinh vt nhn thc
Hnh 11.13: M hnh cu trc ca DNA
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Hnh 11.13b. Nucleotide, DNA v RNA vi sinh vt nhn thc (eukaryote) vt cht di truyn cng c bn cht ha hc l acid nucleic DNA, RNA. Qu trnh t sao chp DNA cng l c s di truyn t th h t bo ny sang th h t bo khc hay th h ny sang th h khc. im khc bit ln gia sinh vt nhn thc v sinh vt nhn s l nhn thc c mng nhn v genome thng gm hai b (lng bi) hoc i khi nhiu hn nhim sc th ( thc vt. tam bi, t bi, a bi) cng cu to t DNA xon kp nhng khng khp kn (c u t do). Bn cnh genome nhim sc th ( trong nhn), cc eukaryote cn c h thng tn hiu di truyn khc l DNA trong cc ty th v lp th. Genome ca cc tiu c quan (ty th v lp th) c phin m thnh RNA thng tin tng ng vi qu trnh din ra tng t t bo nhn s. Gene nhy (transposon) l on DNA c bit gp c sinh vt nhn s v sinh vt nhn thc, khng c v tr n nh trong nhim sc th. on ny c th tch ri khi v tr vn c ca n v di chuyn n mt v tr khc ca cng nhim sc th hoc sang nhim sc th khc. Khi mt gene nhy chn vo khong gia gene khi ng (promoter) v gene cu trc cng nh chn vo gia gene cu trc lm cho qu trnh phin m khng th khi ng hoc b gin on.
III. C CH CHNG S XM NHP CA GENE L (hn ch v ci bin)

T bo c c ch t sao chp v sa sai bo m cho vt cht di truyn n nh. Tuy nhin trong nhiu trng hp t bo chu s xm nhp ca gene l, nh cc phage xm nhp vo t bo vi khun. Khi t bo cn c c ch khc chng li s xm nhp ca DNA t ngoi vo. S hn ch (restriction) ny l nh cc t bo k ch c h thng enzym nhn bit v ct cc DNA l (gi l cc enzym hn ch - restriction enzym) trong khi cc DNA t bo ch khng b ct l nh s ci bin (modification). Hn ch v ci bin ph bin vi sinh vt v c nghin cu k vi khun. Mt mt chng c tc dng nh du DNA t bo, mt
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khc phn hy DNA l. H thng gm hai hot tnh enzym trn cng mt phn t protein hay trn hai protein ring r. hot tnh endonuclease v methyltransferase. Cc endonuclease ny nhn bit mt trnh t c hiu di ngn khc nhau ty loi enzym v phn ct DNA ny ti v tr hoc cch xa im nhn bit . Do DNA ca mnh c methyl ha ti cc im adenine (ti N-6) v cytosine (ti N-5 v N-4) bi hot tnh methyltransferase tc dng phn ct ca hot tnh endonuclease b tr ngi. Cc endonuclease u ct DNA si i v c th chia thnh hai loi. Loi I nhn bit trnh t DNA c bit v ct DNA ny xa v tr nhn bit. Endonuclease ca phage P1 thuc loi ny. Loi II nhn bit v phn ct DNA trong trnh t nhn bit l mt vng c trnh t i xng quay kp (trnh t i xng o v hay trnh t lp ngc hng - palindrome). Do phn ct ti nhng im c hiu to ra nhng mnh DNA xc nh nn enzym hn ch loi II c s dng trong k thut ti t hp v to dng gene. Bng di y gii thiu mt s enzym hn ch. Ch mt s enzym hn ch (Eco RI, Hha I) ct DNA hai si to ra u gm mt s nucleotide mt si (u dnh - cohesive), trong khi mt s enzym hn ch khc (Ba II, Hae III) ct DNA hai si to ra u bng (u ty). Bng 10.4. Mt s enzym hn ch, dng vt ct v ngun gc ca chng
Enzym Eco RI Hha I Ba II Hae III Bam HI Hind III Kpn I Mbo I Trnh t im nhn bit 5'-GAA*TTC 5'-GC*GC 5'-TGGC*CA 5'-GG*CC 5'-GGATCC 5'-AAGCTT 5'-GGTACC 5'-GATC Ngun gc Escherichia coli RY 13 Haemophilus haemolyticus Brevibacterium albidum Haemophilus aegypticus Bacilus amyloliquefaciens H Hemophilus influenzae Rd Klebsiella pneumoniae Moraxella bovis
Hnh 11.14 . Hai dng u ct ca enzym hn ch loi II.

(a1) ct to u ty (ct thng) v (a2) to u so le vi trnh t 6 nucleotide.(b1)ct Hnh thnh u ty (ct thng) v (b2) to u so le vi trnh t 4 nucleotide
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IV. BIN D
1. Bin d kiu hnh
Bin d kiu hnh cn gi l thng bin, l hin tng c s khc bit v cu trc cng nh c im pht trin gia th h con ci so vi th h b m cng nh gia chng vi nhau di tc ng ca mi trng v khng lin quan n s thay i vt cht di truyn. y l nhng bin d tm thi, thun nghch v khng n nh ca ton b qun th sinh vt. Nhng bin d ny xut hin chm v mt i khi cc yu t lm xut hin chng mt i. Nhiu nm gy bnh thng biu hin hai dng hnh thi khc nhau, thng gi l nm nh hnh. Chng c dng hnh cu (dng nm men) khi trong ni quan ng vt mc bnh nhng c dng si khi nui cy trn mi trng nui cy nhn to hay mi trng ngoi, v d, Aspergillus fumigatus, Coccidoides immitis, Histoplasma farsimosa,... Bin i hnh thi do thng bin cn biu hin dng khun lc. Nhng vi khun cng loi khi pht trin trn mi trng c c th hnh thnh khun lc lng (dng S) hoc nhm (dng R) hoc nhng dng trung gian.
2. Bin d kiu gene

Khi nim bin d kiu gene sinh vt a bo thng c coi l ng nht vi khi nim t bin (mutation). Tuy vy, vi sinh vt, thng l n bo, th khi nim bin d kiu gene phi c hiu rng hn v nh l s hnh thnh cc bin chng c th l kt qu ca t bin (bin i cu trc genome sn c) hoc ca qu trnh tip nhn vt cht di truyn t bn ngoi (thng tip theo sau l qu trnh ti t hp thng tin di truyn vo genome nhim sc th). 2.1.Th nghim Luria v Delbrruck (1943) 2.2.Phng php pht hin chng t bin bng th nghim in vt ca Ledergberg, 1952
H.10.15a.
Tm tt th nghim in vt ca Ledergberg, 1952
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2.3. Bn cht ca t bin: B nucleotide ca DNA thuc mt gen l mt mt m thng tin, n tng ng vi cu trc mt protein c hiu. Mi thay i ca b ny dn n s thay i cu trc v chc nng ca protein , hnh thnh mt tnh trng t bin. t bin l nhng thay i tm phn t. Mi t bin d l ngu nhin hay do cm ng th s thay i ca b nucleotide c th xy ra hai kiu: thm, bt hoc thay th mt i baz bng mt i baz khc (t bin im) hoc thm, bt hoc thay th mt on DNA bng mt on DNA khc (t bin on). Mc d t bin on thng kh gp, nh hin tng khuych i gen, nhng do tnh ph bin ca t bin im nn ngi ta thng ng nht khi nim t bin im vi t bin. t bin ny thng khng bin o. t bin im tuy nh nhng thng gp phn quan trng trong qu trnh tin ha. Khi mt baz b mt hoc c chn thm vo trnh t ca gen cu trc s xut hin s thay i mnh m trong cu trc protein do thay i ton b trnh t Acid amin trong thnh phn chui polypeptide k t v tr thm im hay mt im (c theo hng 5'3' ca RNA thng tin). Bin i ny c gi l s chuyn dch khung (frame shift). Trong a s trng hp, c bit t bin im v tr u 5' ca si dng ca gen cu trc, cng l u 5' ca RNA thng tin, nhng bin chng ny thng mt kh nng sng nu bin d xy ra trn gen thit yu (house-keeping genes) ca sinh vt do cu trc (ha hc cng nh khng gian) protein thay i dn n mt chc nng vn c (cu trc, enzym, kch thch, th th,...) ca n. t bin th im (GCAT v ngc li) ngc li l t bin thng gp nhng t biu hin ra bn ngoi v thng kh pht hin. Khi DNA gen c phin thnh RNA thng tin, im t bin th im khng lm thay i khung c ca phn ln chiu di ca chui RNA thng tin. V vy thnh phn ca chui polypeptide ch thay i ti mt v tr tng ng vi im t bin trn gen. Khi , nu acid amin mi khc nhiu so vi acid amin nguyn thy v thuc tnh phn cc, tnh i thy v tnh phn ng th tnh cht nguyn thy ca protein b thay i dn n chc nng c ca protein b mt. Hin tng ny biu hin trm trng nu acid amin thay th nm trung tm hot ng ca mt enzym thit yu, kt qu c th l bin chng khng th tn ti. Trong trng hp acid amin thay th khng c s khc bit nhiu so vi acid amin nguyn thy hoc t bin to mt m thoi ha (nhiu m b ba nucleotide cho mt acid amin) th t bin khng biu hin tnh trng v tch ly ln trong qun th. y thng l nguyn nhn ca s hnh thnh loi mi trong qu trnh tin ha. Chng loi t bin thng gp kh phong ph. C th gp t bin hnh thi nh t bin dng khun lc, thay i kh nng sinh sc t hoc kh nng hnh thnh nha bo. c bit c th gp cc t bin khuyt dng hay t dng (auxotrophic mutants): cc chng t bin ny mt kh nng pht dc nu trong mi trng thiu mt yu t no . Ngc li, mt s t bin dn n vic tng kh nng tng hp cc yu t sinh trng hay cc hp cht khc nh acid amin,... (ng ngha vi vic mt kh nng iu tit sn xut tha). Cc t bin khc cng thng gp vi khun l t bin thay i kh nng ln men, kh nng s dng cc ng, t bin gy vng bn i vi cc nhn t c ch dn n thay i c lc, thay i tnh khng nguyn hoc s mn cm i vi khng sinh hoc tia t ngoi, v.v...
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IV. C CH VN CHUYN VT LIU DI TRUYN

C ba c ch vn chuyn di truyn khc nhau vi khun. Bin np (cn gi l chuyn th - transformation), ti np (transduction, cn gi l chuyn np) v tip hp (conjugation, cn gi l giao np hay giao phi).
1. Bin np (Transformation)
Bin np (hay chuyn th) l s vn chuyn thng tin di truyn nh s xm nhp ca cc phn t DNA t do (c ngun gc t t bo cho phn gii) vo t bo nhn lm cho genome ca t bo nhn thay i. Kt qu ca bin np ph thuc vo mc tng ng trong cu trc phn t DNA ca t bo cho v t bo nhn. V vy s bin np thng c kt qu hn trong gii hn mt loi c cc c th c nhng tnh trng khc nhau. Th nghim 1
Th nghim 2
H.10.15b. Tm tt th nghim ca Avery, MacLeod v McCarty, 1944 iu kin bin np. Qu trnh bin np ch xy ra trong khi cc vi khun nhn mt tnh trng sinh l c bit ( khong 1 - 15% t bo qun th, thng thng giai on pht trin gim). Tnh trng ny lm thay i cc tnh cht b mt t bo nhn c cc yu t cm ng enzym c bit trn cc on phn t DNA c th xm nhp c. Trn b mt t bo nhn c cc yu t cm ng cho on DNA c th c nh. C chng 30 - 70 vng cm ng nh th. on DNA nhp vo t bo c tch i, v mt mch n c ghp vo
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nhim sc th ca t bo nhn. Qu trnh ny c gi l s ti t hp (recombination). Tn sut ti t hp ph thuc vo c tnh cc on nhim sc th khc nhau, vo s tng ng cc phn t DNA. Bin np t nhin thng din ra h Enterobacteriaceae. Trong iu kin th nghim tn sut bin np tng cao nh t bo c x l lm thay i mng t bo cht (nh x l polyethylene glycol) v gy sc nhit (chuyn t bo t nhit 0C sang nhit 40 - 50C trong vng 50 - 60 giy). Frederick Griffith (1871 - 1941) l ngi u tin pht hin s bin np ph cu khun (Streptococcus pneumoniae, tn c l Diplococcus pneumoniae hay pneumococcus) vo nm 1928. Vi khun ny c hai dng khc nhau. Dng gy bnh cho ng vt hnh thnh khun lc lng k hiu l S. Dng khng gy bnh hnh thnh khun lc nhm k hiu R. Nu tim vi khun sng chng S th lm chut nht cht, tim S cht do qua x l nhit (h nng) th chut vn sng, cn tim vi khun chng R sng cho chut th chut vn sng, nhng tim R sng ng thi vi tim S cht li lm chut cht v sau trong mu chut cht phn lp c chng S c trng. Griffith cng nh cc nh khoa hc thi by gi cha hiu ngha su xa ca s kin. Avery, Mac-Leod, Mac-Carthy nm 1944 phn lp c nhn t bin np v xc nh l DNA. Nhng lc by gi gii khoa hc cha chp nhn DNA l cht liu di truyn. n nm 1952 lc Hershey v Chase chng minh DNA ca Phage i vo bn trong vi khun trong khi v Protein b gi bn ngoi th DNA mi c cng nhn l nhn t bin np. S bin np c khm ph thm nhiu vi khun khc: Hemophilus, Neisseria, Streptococcus, Bacillus, Acinetobacter... ngha ca s bin np: S bin np cho php kt hp vo nhim sc th vi khun DNA tng hp hoc DNA bin i invitro. Bin np cng c s dng xc nh nhng vng rt nh trn bn di truyn ca vi khun. Bin np c th xy ra trong thin nhin. Ngi ta chn lc vi khun ti t hp c lc tng lc tim 2 chng ph cu vo phc mc chut nht. Nh th s bin np c mt ngha dch t hc, tuy nhin s vn chuyn di truyn bng bin np khng hu hiu bng s vn chuyn di truyn bng plasmid.
2. S ti np (Transduction)
Ti np l s vn chuyn ADN t vi khun cho n vi khun nhn nh phage. Trong ti np chung, phage c th mang bt k gen no ca vi khun cho sang vi khun nhn. Trong ti np c hiu, mt phage nht nh ch mang c mt s gen nht nh ca vi khun cho sang vi khun nhn. 2.1. S ti np chung Kiu ti np ny c Zinder v Lederberg khm ph nm 1951 lc kho st s tip hp Salmonella. Lc lai ging chng L22 v chng L2, mi chng mang mt cp tnh trng t bin khc nhau th thu c vi mt t l nh mt s t bo ti t hp mang mt cp tnh trng t bin nh t bo hoang di gc, phage P22 c tm thy trong nc lc canh khun ca 2 chng vi khun trn. Phage P22 c lc vi chng L2 nhng n ha vi chng L22. Lc cho P22 vo canh khun L2 th tt c t bo ca L2 b dung gii v phng thch phage P22 trong c mt t phage cha mt mnh ADN ca L2. Lc cho chng L22 vo dung dch phage P22 ni trn th mt s phage vn chuyn nhng gen ca L2 vo L22 v vi khun t dng nh type hoang di c pht sinh qua ti t hp.
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2.2 S ti np c hiu Nm 1954 Mores nhn thy phage n ha c th thc hin mt kiu ti np khc gi l ti np c hiu. N ch vn chuyn mt nhm gii hn gen, nhm gen c chuyn nm st ch prophage gn vo nhim sc th. S ti np c hiu vn chuyn nhng gen c hiu nhim sc th v i hi s tch hp ca phage. Hn na khc vi ti np chung, phage ti np c hiu ch c phng thch lc chiu tia cc tm ch khng bao gi t tch ri khi nhim sc th. Lc chiu tia cc tm vo mt canh khun E.coli K12 Gal+ sinh tan vi phage th trong nhng phage c phng thch c khong 10-6 d Gal. dGal l phage mang gen Gal nhng thiu mt dan ADN m n li trn nhim sc th ca vi khun. l phage khng hon chnh khng th nhn ln nu khng b sung an ADN b thiu nhng c th vn chuyn gen Gal. Lc cho dch dung gii ni trn vo canh khun E.coli K12 Gal- th thu c khang 10-6 vi khun ti np galactoza dng tnh. l hin tng ti np tn s thp LTF (Low Frequency Tranduction). Mt s t t bo trong canh khun E.coli K12 Gal- nhim dch LTF tr nn hai ln sinh tan vi phage bnh thng v phage dGal. Nhng t bo ny hnh thnh clon t bo hai ln sinh tan. Lc chiu tia cc tm vo nhng t bo ni trn th thu dc mt dch phage trong nhng phage v phage dGal bng nhau. l s ti np tn s cao (High frequency transduction). ngha ca s ti np: S ti np c th cho php xc nh nhng gen rt gn nhau nhim sc th, do c s dng thit lp bn di truyn. Da vo tn s ti np hai gen cng mt lc ngi ta c th xc nh khang cch gia cc gen v nh th xc nh v tr ca chng. Trong thin nhin s ti np gi mt vai tr c ngha trong ly lan cc Plasmid khng thuc vi khun gram dng nh plasmid penicillinase t cu. Ngai ra prophage c th em li cho vi khun mt s tnh cht c bit quan trng v d prophage trc khun bch hu.
3. Tip hp (hay giao np - conjugation)

Tip hp l hin tng vn chuyn nhng yu t di truyn t vi khun cho (c) sang vi khun nhn (ci) khi hai vi khun tip xc vi nhau. Lederberg v Tatum ln u tin nm 1946 pht hin ra s tip hp vi khun. Hn hp hai bin chng E.coli K12 vi nhau. Chng (1) khuyt dng vi Biotin v Methionin nhng c kh nng tng hp Threonin v Leucin k hiu B-M-T+ L+. Chng 2 khuyt dng vi Threonin v Leucin nhng tng hp c Biotin v Methionin, k hiu B+M+T- L-. C hai bin chngny khng mc c trong mi trng tng hp ti thiu khng cha bn cht trn nn khng to thnh khun lc. Nu trn hai bin chng trn vi nhau ri cy vo mi trng ti thiu th thy mc cc khun lc vi tn s 10-6 . iu ny cho thy hai chng lai vi khun qua trao i gen to thnh cc t bo ti t hp c kh nng tng hp bn cht : Biotin, Methionin, Threonin, Leucin, k hiu B+, M+, T+, L+. Hin tng tip hp lin quan n nhn t sinh sn F. Nhn t F l mt Plasmid. T bo cha F l t bo c hay t bo F+ ng vai tr t bo cho. T bo khng cha F hay l t bo ci F- dng vai tr t bo nhn. Trong th nghim tip hp trn, chng (1) ng vai tr chng cho v chng (2) ng vai tr chng nhn. T cc t bo F+ , Cavalli phn lp c cc t bo Hfr (High frequency of recombination) c kh nng vn chuyn gen vi mt tn s cao. Khi lai Hfr x F- th thu c cc t bo ti t hp 1000 ln nhiu hn khi lai F+x F-.
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Trong cc t bo F+ nhn t F to nn mt lc c bit gi l lc tip hp. Chnh nh lc ny m xy ra s tip hp gia cc vi khun. Trong cc t bo Hfr nhn t F tch hp vo nhim sc th ca vi khun v ch c kh nng sao chp cng vi nhim sc th. t bo Hfr nhn t F cng to nn mt lc tip hp, nhng trong qu trnh tip hp v nm cui b gen, n y b gen vo t bo nhn qua cu nguyn tng ni lin hai t bo tip hp. t bo Hfr trong mt s trng hp F c th tch ri khi nhim sc th v mang theo mt on ADN ca nhim sc th v c gi l F . F c kh nng t sao chp v c th vn chuyn vo t bo nhn. F c th vn chuyn mt s tnh trng ca mt vi khun ny sang mt vi khun khc. F c s dng phn lp v vn chuyn mt s gen chn lc. Tip hp thng xy ra gia nhng vi khun cng loi nhng cng c th xy ra gia nhng vi khun khc loi nh E.coli vi Salmonella hoc Shigella nhng tn s ti t hp thp. ngha ca s tip hp: S tip hp l mt cng c kho st quan trng trong nhiu lnh vc sinh l v di truyn vi khun v n cho php to nn nhng vi khun phi hp nhiu t bin khc nhau. Nh k thut tip hp ngt qung ngi ta thit lp bn nhim sc th ca vi khun. Ngoi ra chng F c s dng nghin cu hot ng ca mt s gen. Trong thin nhin s tip hp gi mt vai tr ng k trong bin d ca vi khun c bit trong ly lan tnh khng thuc gia cc vi khun Gram m.
V. K THUT DI TRUYN
K thut di truyn bt u nh mt khoa hc t 7/1972 khi nhm nghin cu ca Paul Berg (Vin hn lm khoa hc quc gia M) hon thnh vic ch to b gene lai gia virus SV40 ca kh vi phage lambda. Ln u tin kt ni hai virus khc nhau hon ton v mt di truyn thnh mt chnh th v c nhn ln di dng plasmid trong t bo vi khun. Phn t DNA lai cha cc thng tin di truyn ca virus SV40, thng tin iu khin qu trnh t sao ca DNA phage lambda cng nh ton b chc nng ca operon galactosidase ca vi khun E. coli. Ban u khi nim k thut di truyn ch cc k thut ti t hp DNA nhng dn dn ni dung ca thut ng ny m rng ti cc k thut lin quan nh PCR vi cc bin tng PCR, k thut lai phn t (Northern hybridization, Southern hybridization, microarray,...
a b c Hnh 11.16. Virus SV 40. a. nh hin vi in t; b. V capsid; c. DNA
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Nguyn tc chung ca cng ngh DNA ti t hp gm mt s bc. Trc ht DNA genome mc tiu v DNA chuyn ti (vector) c tinh ch v ct thnh on nh mt loi enzym hn ch. Trong nhiu trng hp (c bit i vi sinh vt bc cao) DNA genome mc tiu c thay th bng DNA tng b (cDNA) ca RNA thng tin thnh thc. DNA ny c tng hp t khun RNA thng tin nh s xc tc ca enzym sao chp ngc (reverse transcriptase, RNA-dependent DNA-polymerase). DNA vector c th l ca mt plasmid hoc phage (nu vt mang, dng ha gene l vi khun), plasmid Ti ca vi khun Agrobacterium tumefaciens (nu vt mang dng ha gene l t bo thc vt), l virus (retrovirus hoc adenovirus khng gy bnh, nu vt mang dng ha gene mc tiu trong t bo ng vt). Bc th hai l chuyn vn DNA ti t hp vo c th vt mang, dng ha v bc tip sau l sau nui cy vt mang, sn xut (chit xut v tinh ch) sn phm mc tiu.
a b c Hnh 11.17. Bacteriophage lambda: a. nh hin vi in t; b. V capsid; c. DNA Cho n nay, k thut di truyn t c nhng bc tin vt bc. Nhiu sn phm c ch c sn xut t cc sinh vt ti t hp hay sinh vt bin i gene (genetically modified organisms - GMO) nh ging ng cc, u tng, ht c du, bng,... mang gene chng su bnh hoc khng thuc chng c di hoc c cht lng sn phm nng cao nh khoai ty cha nhiu protein, la giu vitamin A v st,... to ra c nhng vaccine ti t hp c th d sn xut (d nui cy trn la cy t bo hay phi g) v d s dng (cho n, cho ung), hoc cc ng vt ti t hp sn sinh nhng sn phm hu ch nh thuc cha bnh, thm ch to ra nhng n ln mang khng nguyn ngi hy vng c th l vt cho tim hoc ni quan khc iu tr nhng trng hp ngi bnh cn phi thay th ni quan,... Tuy nhin, k thut di truyn cng t loi ngi vo tnh trng c th phi ng u vi nhng e da mi lin quan n sc khe ca ngi v sinh thi - mi trng. Cha th chng minh c tnh v hi ca GMO thc phm i vi con ngi, cng nh cha th ph nhn kh nng lan truyn gene khng thuc (ca plasmid vector) trong tp on cc vi khun t nhin ca c th (v d, trong ng rut) ngi v ng vt. Chm sc nhng loi cy trng khng thuc dit c v thuc dit su bnh c th dn n lm dng nng dc v nu thoi ha tr thnh c di th thc vt khng thuc v khng su bnh s l mt nguy c mi i vi nng nghip. Tng t, nguy c nhng sinh vt GMO ln t sinh vt t nhin l rt cao v c th dn n s tuyt dit ca nhiu loi sinh vt hin c, nh hng xu n mi trng sinh thi,...
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PCR (phn ng chui trng hp. polymerase chain reaction) l mt k thut ring bit c sng to khuych i DNA in vitro da trn c s phn ng t sao ca DNA nh DNA-polymerase t cc deoxyribonucleoside triphosphate lun cn mi (primer) l on oligonucleotide (trong t bo l on RNA do primase xc tc tng hp) gn kt sn mt cch tng b trn khun si n ca phn t DNA. Lai phn t (hybridization) c th gip nhn bit s hin din ca mt phn t c hiu no , gm lai Southern (phn tch DNA) hoc Northern (phn tch RNA) v cc k thut min dch hc nh Western blot analysis v ELISA. Mt trnh t mch n (DNA hoc RNA) bit trc gi l d (probe) c nh du (bng cht phng x) c chun b sn v cho tip xc vi sn phm RNA hoc DNA si n c nh trn gi th (mng nylon hoc mng nitrocellulose), nu c trnh t tng b d s gn kt mt cch c hiu. Sau khi ra k mng gi th, nu pht hin c d (nh k thut phng x t k) trn mng chng t trong sn phm c cha trnh t nucleotide tng b. Western blot analysis l phng php lai khng th gn kt vi mt enzym c kh nng chuyn mu c cht thch hp (nh du bng peroxidase hoc phosphotase kim) vi sn phm c nh trn mng, trong khi ELISA l phng php lai khng th nh du vi sn phm c nh trn b mt l khay. Nu trong sn phm c khng nguyn c hiu th khng th nh du s kt hp c hiu v khng b ra tri. Kt qu l nu pht hin c yu t nh du (nh k thut chuyn mu c cht enzym) th chng t trong sn phm c khng nguyn hay protein c hiu. *Cu hi n tp chng 10 1. Vt cht di truyn sinh vt nhn thc, sinh vt nhn s v virus? 2. S t sao DNA? Qu trnh phin m? Qu trnh dch m? 3. S iu tit phin m v iu tit dch m? 4. Ci bin v hn ch? Bin d kiu hnh? t bin? 5. Bin np (chuyn th) vi khun? 6. Ti np vi khun? Tip hp vi khun? 7. Tng tc di truyn virus" 8. K thut di truyn v cng ngh DNA ti t hp? 9. PCR cn nhng yu t ha hc g, nhng iu kin phn ng nh th no? 10. Cc k thut lai phn t da trn tnh cht c bn g ca cc yu t tham gia phn ng?
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni. 2. Bin Vn Minh (ch bin)2006, Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, Vi sinh vt hc, NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty, 2007, Vi sinh vt hc, NXBGD H Ni.
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* Ti liu tham kho

1. Nguyn Thnh t . 1999. Vi sinh vt hc, NXBGD, H Ni. 2. http.//vietsciences.net v http.//vietsciences.free.fr
* Gii thch thut ng

Bin np l qu trnh chuyn DNA trc tip tch ra t t bo th cho sang t bo th nhn. DNA ny nm t do trong mi trng (dung dch) do mt vi khun (th cho) phng ra. T bo th cho v th nhn c th c bt ngun t nhng sinh vt khc nhau nh: thc vt, ng vt v vi sinh vt. Trong khun kh ca mc ny ch xt hin tng bin np vi khun. Khc vi tip hp v ti np, bin np khng cn s tip xc trc tip gia 2 t bo cng nh khng cn vt trung gian nh cc phage. Bin d t bin l nhng bin trong vt cht di truyn (nhim sc th, DNA) dn n s bin i t ngt ca mt hoc mt s tnh trng, nhng bin i ny bm sinh v di truyn chi cc i sau. DNA si n (ssDNA) v c ch sa li t bin Trong mt s loi virus, DNA nhim sc th ca chng c dng si n, khng cun xon. V rng hu ht cc c ch sa cha sai hng DNA ca t bo ch thc hin trn cc base bt cp, do cc virus mang DNA genome si n thng c tn s t bin cao hn cc loi khc. Nh , nhng loi virus ny thng c kh nng thch nghi nhanh hn trch b loi b, hay tuyt chng. Tuy nhin, c im ny khng t ra c ch i vi nhng sinh vt cu to phc tp hn v c tn s sinh sn chm hn so vi virus. iu ny gii thch ti sao ch c nhng loi virus mi dng DNA si n lm vt liu di truyn. Ngoi ra, nhng virus DNA si n cng tit kim nng lng tt hn vi c ch nhn i DNA so vi nhng loi virus DNA si kp. DNA vng: Trong t nhin, cng nh iu kin in vitro, phn t DNA c th tn ti di dng si vng, mch kp. dng cu trc ny, cu trc xon khng gian khng d dng b tho xon do nhit hay cc qu trnh ho hc nu khng lm t gy 1 mch. Trong t nhin, cc topoisomerase thc hin nhim v tho xon bng cch ct tm thi mt mch v gn li sau khi tho xon, qu trnh ny l bc khi u cho hot ng phin m. Trong phng th nghim, ngi ta c th gn ni 2 u ca si DNA mch thng thnh 1 DNA vng trong qu trnh to ra cc plasmid ti t hp. DNA polymerase l mt enzym tham gia chnh vo qu trnh nhn i DNA. Nhng enzym ny xc tc cho qu trnh polymer ha cc deoxiribonucleotide da trn trnh t ca mt chui DNA khc. Si DNA mi c to thnh s b sung vi si DNA khun theo nguyn tc b sung. Tt c DNA polymerase u tng hp DNA theo hng t u 5' n u 3' ca nhim sc th (5' v 3' l v tr ca nguyn t cc bon). Cho ti nay, khng c enzym DNA polymerase no l c kh nng tng hp mt chui mi (de novo) m khng cn nhm OH u 3' c sn (tnh cht ny ch c nhng RNA polymerase). V l do ny m DNA
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polymerase cn n s c mt ca cc primer, t enzym DNA polymerase c th gn cc nucleotid tip theo. Cc primer chnh l cc an oligonucleic gm cc baz ca RNA v DNA trong hai baz u tin lun l baz ca RNA. Cc primer ny c tng hp nh hat ng ca mt enzym khc gi l primase. Enzym helicase cn thit cho qu trnh tho xon ca cu trc xon kp DNA tr thnh nhng si n gip cho qu trnh sao chp DNA din ra theo ng nguyn tc bn bo ton (semiconservative) Mt vi loi virus cng m ha nhng phn t DNA polymerase c bit c kh nng sao chp chn lc DNA ca virus thng qua nhiu c ch khc nhau. Cc retrovirus m ha mt loi DNA polymerase s dng khun l RNA tng hp nhng phn t DNA, l cc enzym phin m ngc (reverse transcriptase). t bin gen l nhng bin i trong s lng, thnh phn, trt t cc cp nucltt, xy ra ti mt thi im no trn phn t DNA (mt, lp, thm, thay th mt cp nucltt). Mi bin i mt cp nucltt no s gy mt dt bin gen. t bin biu hin tng c th ring l, khng tung ng vi iu kin sng, thng l t bin ln v c hi cho c th. Cng c mt s l t bin tri, c ngha trong chn ging v tin ha. Sinh vt bin i gene (genetically modified organisms - GMO) S kt hp v tch ri 2 chui n Cc lin kt hydro gi hai chui xon kp l nhng lin kt yu khin chng d dng c tch ra nh enzym (trong iu kin in vivo) hoc nhit trn 90C (iu kin in vitro, PCR). Nhng enzym nh helicase c chc nng tho xon cc chui cho php cho cc DNA polymerase, RNA polymerase thc hin hot ng. Trong qu trnh tho xon, cc helicase phi ct lin kt phosphodieste ca mt trong hai chui trnh cc chui b xon vng quanh. Dng thng tin trong t bo prokaryote v eukaryote - Prokaryote: DNA - Eukaryote: DNA RNA Protein RNA Protein
pre-mRNA
c im cu trc nhim sc th ca prokaryote - Xon kp: khe nh, khe ln; DNA-binding protein gn vo khe ln - Cu trc bc hai: thn - vng (stem-loop) hay kp tc (hair spin) ni nhn din ca protein iu ha - Cu trc siu xon v cu trc vng m: topoisomerase II v I c im cu trc nhim sc th ca eukaryote - Kch thc ln - Nucleosome, chromatin, nhim sc th - Telomere hai u v centromere gia - Ba nhm DNA:
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+ DNA mt bn sao: m ha protein + DNA lp li trung bnh: m ha histone, immunoglobin, rRNA, tRNA + DNA v tinh: 20% tng DNA, chc nng cha r Cc yu t di truyn (genetic element) - Cc yu t di truyn ngoi NST - B gen virus - Plasmid - B gen ti th - B gen dip lp th - Yu t di ng (IS element), gen chuyn v (transposon) Enzym ct gii hn v s bin i DNA - Enzym ct gii hn: nhn din v ct DNA ngoi sinh ti trnh t i ngu - Enzym bin i: bin i DNA ni sinh ti trnh t i ngu Sao chp DNA (replication) - Tch mch to 2 khun: ORI (origin of replication), topoisomerase, helicase, ch ba sao chp (replication fork) - Sao chp 2 phn t DNA mi bn bo th: + DNA polymerase III, tng hp theo chiu 5 3 + DNA mch vng: hai ch ba sao chp ngc chiu + Mch trc (leading strand, 3 5): tng hp lin tc, hng i vo ch ba (DNA plolymerase III) + Mch sau (lagging strand, 5 3): tng hp cc on Ozaki, hng i ra khi ch ba (primase, DNA polymerase III, DNA polymerase I, ligase) Plasmid l nhng phn t ADN dng vng trn nm ngoi nhim sc th v c kh nng t sao chp. Plasmid c trng khi nh so vi nhim sc th. Hai plasmid gi l khng ph hp nu chng khng th tn ti bn vng trong cng mt t bo. Nhng plasmid F mang nhn t sinh sn F. Nhng plasmid cng tn ti trong mt vi khun vi nhn t F gi l Fi+ nu chng ngn cn s tip hp ca F, gi l Fi- trong trng hp ngc li. Phin m (transcription) - Promoter: RNA polymerase nhn din v gn vo trnh t bo tn -10, -35 trn mch mang m (sense strand, 3- 5) - Sigma tng tc vi RNA polymerase li v DNA, ri khi enzym khi phin m bt u - RNA polymerase tng hp theo chiu 5 3 - Kt thc phin m ti im kt hc (cu trc bc 2) - prokaryote: polycistronic - eukaryote:
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+ Pre-mRNA cha exon v intron + Intron: 5-GUAAG-3, th ct (splicesome) + RNA trong splicesome (ribozyme) ct u 5, 3 v ni hai u M di truyn - 64 m b ba - M khi u: AUG - M kt thc: UAA, UAG, UGA - C ngoi l RNA vn chuyn (transfer RNA, tRNA) - Cu trc bc 2: u mang b ba i m (anticodon), u mang amino acid (aceptor end) - Mi tRNA tng tc vi mt aminoacyl-tRNA synthetase ring - Aminoacyl--tRNA synthetase c trung tm nhn din anticodon v trung tm gn amino acid Dch m (translation) - Ribosome: tiu phn 30S v 50S (prokaryote), 40S v 60S (eukaryote) - Prokaryote: + Tiu phn 30S gn vo trnh t Shine-Dalgano sequence trn u 5 ca mRNA + Shine-Dalgano c trnh t b tr vi u 3 ca rRNA 16S + Dch m polycistronic mRNA - Eukaryote: tiu phn 40S gn vo m 5-cap, dch m AUG u tin - Tiu phn 50S, 60S: + E-site (exit site): v tr cha tRNA khng cn gn amino acid + P-site (peptide site): v tr cha tRNA mang chui peptide + A-site (aceptor site): v tr cha tRNA mang amino acid tip theo - S dch chuyn ca ribosome cn nhn t ko di (elongation factor) v GTP - Ti codon kt thc: Khng c tRNA + Nhn tt phng thch (release factor) nhn din v tr ny, ct peptide khi tRNA + Cc tiu phn ribosome tch nhau v ri mRNA - Khung dch m (khung c, open reading frame ORF) - Gen gi u (overlapping): mt gen c hai khung c (fX174) - Tn sut sai st trong dch m: 10 10
-3 -4
iu ha phin m: kim sot m (negative control) - Lin quan n DNA-binding protein v operator (DNA) - DNA-binding protein l repressor: khi gn vo operator khng to mRNA - Repressor tng tc vi effector:
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+ Repressor-Effector gn vo operator: c ch (repression), arg operon + Repressor-Effector khng gn vo operator: cm ng (induction), lac operon iu ha phin m:kim sot dng (positive control) - Lin quan n DNA-binding protein v activator-binding site (DNA) - DNA-binding protein l activator: khi gn vo operator tng cng to mRNA - Activator tng tc vi effector gn vo activator-binding site, mal operon iu ha ton cc (global regulation) - C ch c ch d ha (catabolite repression) - iu ha nh hng ng thi nhiu gen khc nhau p ng vi bin i ca mi trng - c ch d ha bi glucose E. coli i vi sinh tng hp enzym bin dng cc ng khc: - Protein hot ha d ha (catabolite activator protein, CAP) - CAP tng tc vi effector l cAMP gn vo CAP-binding site vng promoter, xc tin s tng hp mRNA - Nng cAMP ph thuc nng glucose trong mi trng Tng tc gia protein v nucleic acid - Tng tc khng chuyn bit: histone-DNA - Tng tc chuyn bit: protein l dimer, mi monomer gn vo mt trnh t xc nh trn mt si DNA - c im ca trnh t gn protein: lp li o ngc (trn mt mch) t bin (mutation) - t bin, th t bin (mutant)Th t bin c kiu gen b bin i, kiu hnh khng nht thit thay i - K hiu:+ Kiu gen hisC: gen m ha cho protein HisC + Kiu hnh Thr-: mt kh nng t tng hp threonine Khuyt dng (tr dng): auxotroph - Nguyn dng: prototroph C s phn t ca t bin - t bin ngu nhin: sai st trong sao chp, chiu x t nhin t bin nhn to: + Ha cht: cc cht ng dng (analog), cht tc dng trc tip vi DNA Tia UV, tia chiu x ion ha + Gen chuyn v (transposon) + t bin: thay i mt cp base, thm mt hoc mt s cp base, mt on, o on, chuyn onH qu ca t bin: + Lch khung dch m
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+ Mt codon bt u, codon kt thc trn mRNA+ Gen khng c th hin t bin c ch (suppressor) Ti t hp di truyn (genetic recombination) - Da trn s tng ng cao ca hai on trnh t - Xc tc bi RecA - Cc bc: + To mt vt t trn DNA (nicking) + M vng DNA xon kp + Bt cp gia cc on tng ng trn hai phn t DNA mch n + Ct v ni cc mch DNA sao cho trao i on trn cc mch + DNA kiu gen mi c to ra khi hai mch c trnh t khc nhau ngoi vng ct v ni B tr di truyn (complemantation) - Trng hp chng vi sinh vt c mt kiu hnh c quy nh bi mt vi gen, ch mt gen b t bin th kiu hnh c dng t bin - Nu dng t bin c b sung mt an DNA cha cc gen tng ng trong c gen hoang di ca gen t bin th chng t bin s tr li kiu hnh hoang di: hin tng b tr di truyn - Hin tng b tr di truyn: + Ch xy ra khi cc t bin trn b gen v trn DNA ngai lai l khng thuc cng mt gen + B tr v kiu hnh (biu hin gen mc phin m v dch m), khng xy ra s trao i an tng ng (mc DNA) - c ng dng: + Trong chn lc dng ti t hp mc tiu + Xc nh t bin c xy ra cng mt gen trn hai DNA hay khng Bin np (transformation) - Qu trnh t bo tip nhn DNA trn t vo t bo ch -Tnh kh np (competence): c th c tng cng bng x l ha hc, vt l -C ch bin np: + DNA gn ln DNA-binding protein trn vch t bo + Nuclease thy phn mt mch DNA, cho php mch n cn li i vo trong t bo + Mch DNA c mang v bo v bi mt s protein chuyn bit + Mch DNA ti t hp vo b gen bi RecA protein + T bo c kiu gen mi c to thnh khi t bo phn chia Ti np (transduction)
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-DNA ca t bo cho c chuyn qua t bo nhn bi virtTi np chuyn bit (specialized transduction): ti np trn mt s gen nht nh ca vi khun cho (virt mang theo gen ca vi khun khi b ct mt cch khng chnh xc ra khi b gen t bo ch) -Ti np chung (generalized transduction): ti np mt gen bt k t vi khun cho sang t bo nhn (DNA ca t bo b phn on v lp ngu nhin vo v virt mi) -Bin i bi phage (phage conversion): s thay i kiu hnh vi khun do s th hin ca gen virt tim tan Tip hp (conjugation) - Chuyn DNA thng qua giao np vi khun Plasmid xc tin s giao np: + Tng hp khun mao pili gip hai t bo tip xc + To cu giao np (conjugative bridge) truyn DNA + Plasmid sao chp bng c ch sao chp cun vng (rolling circle replication) v chuyn mt bn sao cho t bo nhn + T bo nhn sao chp c plasmid vng mch kp -Yu t F E. coli thc hin vic cho gen trn nhim sc th t bo cho sang t bo nhn (chng Hfr): + S hin din ng thi ca trnh t st nhp (insertion sequence) yu t F v nhim sc th ca t bo + F cha trnh t khi u chuyn (origin of transfer) + Trnh t ny gip F mang theo cc gen ca nhim sc th nm ngay di h lu ca trnh t chuyn t t bo cho sang t bo nhn + S ct khng chnh xc khi sao chp v chuyn yu t F lm tng tn s giao np chuyn bit ca nhim sc th sang t bo nhn Lp bn gen bng cc phng php chuyn gen - Giao np gin on (interrupted mating) c dng xc nh s b trt t ca gen t nhng on DNA -Ti np gin on c dng xc nh chnh xc trt t ca cc gen lin kt cht ch vi nhau (nm rt gn nhau) Gen chuyn v (transposon) - Trnh t st nhp (insertion sequence IS) - Gen chuyn v hay gen nhy (transposon Tn): IS + cc gen khc- Tn s chuyn v ca Tn v IS thp (khong 10-5 10-4)- Cu trc: + Hai u cha cc trnh t lp li (repeated sequence) + Bn trong m ha transposase: nhn din, ct v ni DNA + Chuyn v: gen chuyn v khi gn vo v tr mc tiu s nhn i trnh t v tr mc tiu ny; hai trnh t v tr mc tiu s nm hai bn gen chuyn v
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Di truyn v tnh khng thuc Cc vi khun khng thuc c tm thy khp ni k c nhng ni cha bao gi s dng khng sinh. Ngay khi pht minh penicillin nm 1941, vi khun khng penicillin c bo co. Ngy nay vn khng thuc tr thnh mi bn tm hng u trong iu tr cc bnh nhim trng v khng sinh l mt th nghim hng ngy cc phng xt nghim vi trng. S hnh thnh tnh khng thuc vi sinh vt l do s bin i gen nhim sc th hoc do tip nhn plasmid khng thuc. Vi khun tr nn khng thuc qua 4 c ch: t bin, ti t hp hoc thu hoch plasmid khng thuc hoc thu hoch transposon . a. t bin thnh khng thuc Nhiu th nghim chng minh tnh khng thuc pht sinh do t bin, nh th tnh khng thuc lin h n s bin i gen nhim sc th v c th di truyn cho cc th h sau. S t bin xy ra vi tn sut 10-5 - 10 -9 . Trong tnh khng thuc, khng sinh gi vai tr chn lc ch khng phi vai tr ch o. Vic s dng rng ri khng sinh lm pht trin nhanh chng cc vi khun khng thuc bng cch git cht cc vi khun nhy cm, to iu kin cho vi khun khng thuc pht sinh do t bin chim u th. b.S ti t hp Lc mt t bin khng thuc xut hin mt qun th vi khun th vi khun khng thuc c th vn chuyn gen khng thuc n cc vi khun nhy cm theo mt trong 3 c ch vn chuyn di truyn: bin np, ti np v tip hp ty theo loi vi khun. S ti t hp gia 2 vi khun, mi vi khun khng mt khng sinh lm xut hin nhng vi khun khng vi c 2 loi khng sinh. Trong thin nhin, s vn chuyn gen khng thuc trong ti np v tip hp xy ra vi mt tn sut thp khong 10-5. Trong bin np tn sut cha c bit, nhng c th cn thp hn na. c. Thu hoch plasmid khng thuc Hai c ch khng thuc trn lin quan n gen khng thuc nm trn nhim sc th. y tnh khng thuc lin h n plasmid nm ngoi nhim sc th. Plasmid khng thuc c vn chuyn theo nhng c ch khc nhau ty theo vi khun Gram dng hoc vi khun Gram m. * Plasmid khng thuc vi khun Gram m Nm 1946 ln u tin Nht Bn ngi ta phn lp nhiu chng Shigella khng vi nhiu khng sinh : streptomycin, chloramphenicol, tetracyclin, sulfonamit. Nhn t chu trch nhim v tnh khng nhiu thuc l mt plasmid khng thuc goi l nhn t R. Nhn t R in hnh l mt plasmid ln vi 2 phn, chc nng ring bit. Phn th nht RTF gi l nhn t vn chuyn khng (Resistance transfer factor), khong 80 kb cha nhng gen ca s t sao chp v ca s tip hp. Phn kia nh hn l quyt nh khng (R determinant), kch thc rt thay i v cha gen khng thuc (R genes) RTF v quyt nh khng thng thng to nn mt n v. Nhn t R thng ti t hp vi nhau lm pht sinh nhng t hp mi v khng thuc. Nhn t R khng nhng c th ly truyn rng ri trong nhiu loi vi khun ng rut m cn c th ly truyn cho nhiu vi khun khc nh vi khun t, vi khun dch hch... Nhn t R ly truyn trong vi khun qua tip xc nn tnh khng thuc lan trn trong mt qun th vi khun nhy cm nh mt bnh truyn nhim .
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* Plasmid khng thuc vi khun Gram dng S khng ca nhiu chng t cu vng vi penicillin, erythromycin v nhiu khng sinh cng do plasmid gy nn. Plasmid vi khun Gram dng khng th vn chuyn bng tip hp m bng ti np qua trung gian ca phage. Hin nay phn ln nhng chng t cu khng penicillin bnh vin u mang plasmid penicillinase. d.Thu hoch transposon Transposon cn gi l gen nhy l nhng on ADN, kch thc nh cha mt hay nhiu gen c 2 u tn cng l nhng chui nucleotic ging nhau nhng ngc chiu nhau (inverted repeat). c th nhy t plasmid vo nhim sc th hoc t nhim sc th vo plasmid hoc t plasmid n plasmid. Tt c mi gen trong c gen khng thuc c th nm trn transposon. Nhng transposon ch o s khng kim loi nng, s to thnh c t v kh nng s dng mt s cht chuyn ha (lactose, raffinose, histidin, hp cht lu hunh) c m t. c bit quan trng trong vi sinh hc l nhng transposon khng khng sinh nh Tn3 mang gen khng ampicillin, Tn5 mang gen khng kanamycin, Tn10 mang gen khng tetracyclin, Tn4 mang gen khng ba khng sinh ampicillin, streptomycin v sulfamit.
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CHNG 12 VI SINH VT GY BNH, MIN DCH HC V CC CHT C HOT TNH SINH HC I. VI SINH VT GY BNH
1. Bnh truyn nhim v khng truyn nhim
1.1. Khi nim a. Bnh truyn nhim l bnh do vi sinh vt gy nn, c tnh cht ly lan mnh lm cht nhiu sinh vt trong mt khong thi gian.V d: Bnh u ma ngi, bnh dch t tru b, bnh ln ng du... Nhng bnh khng truyn nhim cn gi l bnh thng thng th khng c kh nng ly lan v khng lm cht nhiu sinh vt. Bnh thng thng pht sinh do thi tit, do v sinh km.V d: Bnh say nng, bnh khng tiu sinh hi...B mt ca bnh truyn nhim c cc nh khoa hc pht hin, do 3 yu t sau y quyt nh: Tc nhn gy bnh (Vi khun, vi nm, VNS hoc virus); Tnh cht phn ng ca c th v yu t hon cnh . b. Bnh c hiu v bnh c hi: Bnh c hiu (Bacteria Phthogene Specifique) l loi bnh cp (c tc nhn gy bnh) cn cha ngay nh dch hch, th t, bnh lao, giang mai, l, thng hn..Bnh c hi (Bacteria Pathogene Opportuniste) thng l do c th b yu, min dch gim, mi trng thay i th nhng VSV vn sng hoi sinh s tr nn gy bnh. 1.2. Ngun gc bnh truyn nhim - Bn ngoi: + Ngi truyn cho ngi: Bnh nhn t trc lc thy r cc triu chng cho ti sau khi khi bnh mt thi gian vn c th mang vi khun gy bnh v c th lm ly sang ngi khc. + ng vt truyn cho ngi: Nh bnh dch hch, bnh di, bnh than... - Bn trong: C mt s vi khun bnh thng, vn sng trn da hay trn c th ngi hay ng vt m khng gy nn bnh. Nhng lc c th suy yu, sc khng gim th chng pht trin mnh v gy bnh. Hoc khi vi khun i ni c tr m gy nn bnh. V d trc khun E.coli i trng th khng gy bnh nhng khi bng quang th li gy bnh. 1.3.Phng thc truyn nhim - Truyn nhim do tip xc: Ngi hoc vt kho tip xc trc tip vi ngi hoc vt m bng s c xt (bnh vim hch nga), giao hp (bnh giang mai, lu, AIDS), b (l mm long mng), lim , cn (bnh di)...s truyn bnh cho nhau. Cng c th do tip xc trc tip vi dng, dng c m mc bnh nh bnh mt ht, bnh un vn... - Truyn nhim do ng tiu ho: Nhiu bnh ng rut ly truyn do n phi thc n c cc vi khun ca ngi bnh bi tit ra (t, l, thng hn, bi lit...)
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- Truyn nhim do ng h hp: Ngi hoc ng vt c th mc bnh do vi khun xm nhp vo c th bng ng h hp nh bnh lao, bch hu, ho g, si... - Truyn nhim do cn trng tit tc: V d nh mui truyn bnh vim no, st xut huyt; b cht truyn bnh dch hch; chy rn truyn bnh st pht ban... 1.4. c im qu trnh bnh truyn nhim Mi loi VSV gy bnh ch gy mt bnh truyn nhim nht nh. Qu trnh bnh truyn nhim gm giai on bnh, giai on tin pht, giai on ton pht v kt thc. Cc thi k th hin s u tranh gia mm bnh v c th. Do tng quan lc lng gia mm bnh v c th, bnh truyn nhim c th biu hin th qu cp tnh lm sinh vt cht nhanh, hoc th cp tnh, th mn tnh hay th n.... C bnh truyn nhim chung cho cc loi vt nui, c bnh chung cho c ngi. Phn ln cc bnh truyn nhim sau khi khi bnh s c min dch trong mt thi hn khc nhau tu theo tng loi bnh.
2. c t v c lc
2.1. c lc L sc gy bnh ca mt chng VSV c kh nng gy bnh nhiu hay t, nng hay nh. nh gi c th c lc ca mt chng VSV, ngi ta biu th bng n v o lng l liu cht ti thiu MLD (mt lng VSV nh nht c th git cht sc vt th nghim trong thi gian nht nh). c lc ca mt chng VSV khng phi l c nh. Khi mi phn lp c bnh phm, VSV thng c c lc cao, nhng qua qu trnh cy truyn th c lc gim dn v c khi mt hn. c lc ca VSV bao gm c t v cc cht khc do VSV sinh ra. 2.2. c t L sn phm ca qu trnh chuyn ho dinh dng ca VSV, gm 2 loi: Ngoi c t v ni c t. Ngoi c t (Exotoxin): l nhng cht c do VSV tit ra ngoi khi VSV cn sng. (un vn, bch hu, c tht...). Ngoi c t d tan v d lan rng, d b cc enzim v ho cht ph hu lm mt c tnh. Thnh phn ho hc l protein nn khng chu c nhit cao. Ni c t (Endotoxin): l thnh phn ca t bo VSV, c tc dng gy c khi t bo VSV ang b dung gii.Thnh phn ho hc l mt phc hp protein-lipid-glucid, chu c nhit cao, c tnh yu, tnh khng nguyn yu, khng c kh nng tr thnh gii c t.
3. Bnh do virus, vi khun v cc phn t hu c khc.

3.1. Bnh do vi khun T cu khun gy bnh
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Phy khun t Trc khun ng rut Cc vi khun gy bnh khc thng gp Cc vi khun khng axit 3.2. Bnh do vi nm Candida albicans gy bnh nm su Piricularia oryzae gy bnh o n Helminthosporum oryzae gy bnh tim la Fusarium fujikuroi gy bnh la von. 3.3. Bnh do virus Bnh cm Bnh si tr em Bnh bi lit Bnh SARS Bnh cm g H5N1
TT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
*a *b
Mt s bnh thng gp St hi quy Phong (hi) Thng hn Lao T Bch hu Un vn Vim mng no L Ng c tht u ma Dch hch St vng Giang mai Bi lit Cm Vim no Nht Bn AIDS Bnh c khoai ty hnh thoi Bnh no (b in -BSE)
Vi sinh vt gy bnh XT Borrelia recurrentis VK Mycobacterium leperae VK Salmonella typhi VK Mycobacterium tuberculosis VK Vibrio cholera VK Corynebacterium diphtheriae VK Clostridium tetani VK Neisseria meningitidis VK Shigella spp. VK Clostridium botulinum Virus u ma VKYersinia pestis Virus st vng VK Treponema pallidum Virus bi lit Virus cm Virus vim no Nht Bn Virus HIV Viroid Prion
Nm pht hin 1868 -1873 1873 -1874 1880 1882 1883 1883 -1884 1884 1887 1891 -1898 1896 1892 -1906 1896 1901 1905 1908 -1909 1993 1934 -1938 1981 1971a 1982b
Thnh phn khng c protein m ch c mt axit nucleic gm khng qu 150nucleotit (1982. Stanley B. Prusiner, M, pht hin nhng ht protein c tnh cht ly truyn, gi l prion, mt tc nhn gy nhim trong mt s bnh no, nh bnh b in).
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II. KHNG NGUYN, KHNG TH V B TH

1. Khng nguyn
1.1. Khi nim Khng nguyn l phn t kch thch p ng min dch ca c th, c bit l sn xut khng th. Thng thng khng nguyn l mt protein hay mt polysaccharide, nhng n cng c th l bt c loi phn t no, mang cc phn t hapten nh v gn vi mt protein chuyn ch. 1) Cc loi khng nguyn a. Min dch nguyn - khng nguyn loi ny kch thch p ng min dch khi c a vo trong c th. Min dch nguyn lun lun l mt i phn t (protein, polysaccharide). Kh nng kch thch p ng min dch ca n ph thuc vo tnh l i vi vt ch, kch thc phn t, thnh phn ho hc v tnh khng ng nht (th d nh phn t protein cha nhiu loi amino acid khc nhau). b. Dung np nguyn - khng nguyn loi ny kch thch tnh trng khng p ng min dch c hiu, do hnh dng phn t ca n. Khi thay i hnh dng, n c th tr thnh min dch nguyn. c.D ng nguyn - y l cht gy phn ng d ng. Chng c th xm nhp do c n, ht, tim vo c th hoc tip xc vi da. 2) Ngun gc ca khng nguyn a. Khng nguyn ngoi sinh l khng nguyn xm nhp vo c th t bn ngoi, nh do ht, n, tim. Bng qu trnh nhp ni bo hoc thc bo, cc khng nguyn ny c a vo t bo trnh din khng nguyn (APC) v c x l thnh cc mnh nh. b. Khng nguyn ni sinh l cc khng nguyn c sn xut bn trong t bo, l kt qu ca qu trnh chuyn ho t bo bnh thng, hoc do nhim khun ni bo hay nhim virus. * C mt ngoi l khng thuc ngoi sinh ln ni sinh, c gi l trnh din cho. c. Khng nguyn khi u l cc khng nguyn c trnh din bi cc phn t MHC I trn b mt t bo khi u. Khng nguyn khi u cng c th c b mt khi u dng, chng hn l, mt th th b t bin. Trong trng hp ny chng b nhn din bi t bo B. 1.2. T bo trnh din khng nguyn (ting Anh l antigen presenting cell, APC) L nhng t bo biu hin khng nguyn l lin kt vi phc h ph hp t chc (major histocompatibility complex - MHC) trn b mt ca n. T bo lympho T c th nhn din c phc hp ny bng cch s dng th th t bo T (T-cell receptor - TCR). 1.2.1. Phn loi t bo trnh din khng nguyn - C 3 loi t bo trnh din khng nguyn chuyn ho chnh: + T bo tua (Dendritic cells)
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+ i thc bo (Macrophages) + T bo B (B cells) Cc t bo tua vi ph trnh din khng nguyn cc ln chnh l nhm t bo trnh din khng nguyn quan trng nht. Cc t bo tua c hat ha lun lun biu hin nhng phn t ng kch hat (v d nh phn t B7) c kh nng hot ha cc t bo T b tr. T bo B, vi nhng khng th trn b mt, c th trnh din rt hiu qu cc khng nguyn c hiu vi khng th, nhng li khng hiu qu i vi cc khng nguyn loi khc. Ngoi ra, mt vi dng t bo bit ha mt s c quan no (v d: vi t bo thn kinh m (microglia) trong no, t bo Kuppfer gan) vn c ngun gc t i thc bo cng c kh nng hot ng nh cc APC. 1.2.2. S tng tc vi t bo T Sau khi nut cc tc nhn gy bnh, t bo tua hoc i thc bo di chuyn n hch lympho, ni cha hu ht cc t bo T. c th n c hch lympho, cc APC di chuyn theo nng cc chemokine trong mu (c ch ha hng ng). Trong khi di chuyn, t bo tua tr nn mt kh nng thc bo v tng cng biu hin cc cht ng kch hat (tr thnh t bo APC chn) tng tc vi t bo T. Cc t bo ny c kh nng thc bo. Chc nng quan trng hn ca nhng t bo APC l chng hot ho t bo T non (naive T cell) thng qua nhng phn t ng kch hot c biu hin trn b mt t bo.
2. Khng th
Hnh 12.1: B mt mt phn t IgG 2.1. Khi nim
Hnh 12.2: Cu trc ca mt phn t khng th.
Khng th l nhng protein c tng hp nh cc t bo lympho. Cc khng th c th tn ti nh cc phn t t do trong dch th hoc di dng phn t nm trong mng sinh cht ca cc t bo limpho.
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Khng th (ting Anh: antibody) l cc phn t immunoglobulin (c bn cht glycoprotein), do cc t bo lympho B cng nh cc t bo (bit ha t lympho B) tit ra h min dch nhn bit v v hiu ha cc tc nhn l, chng hn cc vi khun hoc virus. Mi khng th ch c th nhn din mt epitope khng nguyn duy nht. Cc domain hng nh
Hnh 12. 3: S cc chui ca mt khng th. Phn t khng th cu to t 4 chui polypeptide, gm hai chui nng (H, heavy, ting Anh, mu tm trong hnh 113) ging ht nhau v hai chui nh (L, light, ting Anh, mu xanh l trong Hnh 12.3) cng ging ht nhau. C hai loi chui nh (kappa) v (lambda), do hai chui nh ca mi phn t immunoglobulin ch c th cng l hoc cng l . Cc chui ca immunoglobulin lin kt vi nhau bi cc cu ni disulfide v c n hi nht nh (Hnh 12. 2 v 11.3). Mt phn cu trc ca cc chui th c nh nhng phn u ca hai "cnh tay" ch Y th rt bin thin gia cc khng th khc nhau, to nn cc v tr kt hp c kh nng phn ng c hiu vi cc khng nguyn tng ng, iu ny tng t nh mt enzym tip xc vi c cht ca n. C th tm so snh s c hiu ca phn ng khng th-khng nguyn vi kha v cha kha. Cc domain hng nh (C, constant, ting Anh) c trng bi cc chui amino acide kh ging nhau gia cc khng th. Domain hng nh ca chui nh k hiu l CL. Cc chui nng cha 3 hoc 4 domain hng nh, ty theo lp khng th CH1, CH2, CH3 v CH4. Cc domain hng nh khng c vai tr nhn din khng nguyn, chng lm nhim v cu ni vi cc t bo min dch cng nh cc b th. Do , phn "chn" ca ch Y cn c gi l Fc (tc l phn hot ng sinh hc ca khng th F: fragment, c: cristallisable). Cc domain bin thin Mi immunoglobulin c 4 domain bin thin (V, variable, ting Anh) u tn hai "cnh tay" ca ch Y. S kt hp gia 1 domain bin thin trn chui nng (VH) v 1 domain bin thin trn chui nh (VL) to nn v tr nhn din khng nguyn (cn gi l paratope). Nh vy, mi immunoglobulin c hai v tr gn khng nguyn. Hai v tr ny ging nhau nh c, qua mt khng th c th gn c vi 2 khng nguyn ging nhau. Hai "cnh tay" ca ch Y cn gi l Fab (tc l phn nhn bit khng nguyn, F: fragment, ab: antigen binding). Domain khng nguyn ni gn vo khng th gi l epitope.
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Cc domain s d gi l bin thin v chng khc nhau rt nhiu gia cc khng th. Chnh s bin thin a dng ny gip cho h thng cc khng th nhn bit c nhiu loi tc nhn gy bnh khc nhau. C ch to nn s bin thin ny s c cp nhng phn sau. Isotype iu gy ch l khi gy p ng min dch chut bng albumine ca 1 ngi, khng th sinh ra c tnh c hiu i vi albumine ca bt c ngi no, ch khng ring g ca c nhn ngi ni trn. Nh vy c mt ci g chung cho c mt loi. Nh min dch hc ngi Php Jacques Oudin ra khi nim isotype ch c tnh khng nguyn chung ca loi. Isotype l tr ngi ln cho huyt thanh liu php (th d dng huyt thanh nga c cha khng th khng c t un vn cha bnh un vn cho ngi), cch khc phc l dng cng ngh sn xut cc khng th n dng. Immunoglobulin ngi chia lm 5 isotype, s c trnh by mt phn sau. c tnh isotype khng th c quy nh bi cu trc thuc phn hng nh ca i phn t khng th (c th l trn cc domain CH). Ghi ch: iso=ng, cng isotype: khng nguyn chung ca 1 loi. Allotype Khng hn l immunoglobulin lc no cng c dung np mt c th khc cng loi, Oudin ra khi nim allotype khi quan st thy mt s th li sinh khng th chng chnh cc immunoglobulin th. Allotype cng thuc phn hng nh ca immunoglobulin. Tuy nhin, s khng tng hp do allotype c bit n nhiu nht khng phi l cc immunoglobulin m l cc nhm mu v h HLA. Ghi ch: allo=t ch allele allotype: khng nguyn ca cc c th cng loi mang allele khc nhau. Idiotype Allotype l ranh gii gia hai c th cng loi. em mt khng th th khng albumine ngi (sau y gi l anti-albumine hay Ig1) tim cho mt con th khc cng nhm allotype, ngi ta thy con th th 2 ny li sn xut khng th Ig1 ni trn. Do nhng khc bit v isotype v allotype c loi tr (cng loi, cng allotype), i tng ca vic sinh min dch ny c kt lun l vng c hiu ca khng th 1 k trn. Cu trc to nn tnh c hiu vi khng nguyn c gi l "c tnh idiotype". Khng th anti-albumine gi l idiotype, cng chnh Oudin ngh thut ng ny. Tuy nhin idiotype ch danh chnh l vng bin thin trn khng th (cng nh trn TCR) c hiu vi mt khng nguyn, cn v tr lin kt vi khng nguyn gi l paratope. Ngi ta thnh cng trong vic ct cc idiotype ra khi khng th, phc v nghin cu v pht hin ra khi nim "dy (hay dng thc) idiotype": 1. em mt khng nguyn X gy min dch chut A, ngi ta thu c khng th (idiotype) anti-X (tm gi l Ig1).
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2. Ly Ig1 tim cho chut B (ging ht v di truyn vi chut 1), khng th anti-antiX c to ra (Ig2). 3. Ngi ta chng minh c rng trong cng mt c th, khi tip xc vi 1 khng nguyn X, khng phi ch c 1 Ig1 (anti-X) c sn xut, m l mt dy nhng Ig nh sau: Khng nguyn (X) Ig1 (anti-X) Ig2 (anti-anti-X) Ig3 (anti-anti-antiX)... t nht trong mt s trng hp, Ig3 rt ging (nu khng ni l ging ht) Ig1 v kh nng nhn din khng nguyn X u dy, nh vy, Ig2 c nhng cu trc gy p ng to Ig3 c hiu vi X, ngi ta ni Ig2 hot ng nh mt hnh nh ni ti ca khng nguyn. Ig2 c ng dng nh mt vc-xin (vc-xin anti-idiotype). Suy rng ra, khi mt trong mun vn nhng khng nguyn "tim nng" ca th gii xung quanh xm nhp c th, h min dch to ra mt dy, hay "mng li" idiotype nh cch dng ca Niels Jerne . S phn bit gia "ta" v "khng ta" rt tinh t, khng phi l mt s phn bit trng en hai na mt cch n gin, m l mt h thng cn bng ng trng k, vai tr gy p ng min dch ca khng nguyn l lm xo trn s cn bng . Ghi ch: idio=ring, t mnh idiotype: cu trc ring bit trn phn bin thin ca mi immunoglobuline hay TCR, c hiu cho mt epitope khng nguyn nht nh. T khng th IgM: IgM to nn cc polymer (poly = a, nhiu) do cc immunoglobulin lin kt vi nhau bng cc cu ni ng ha tr disulfide, thng l vi dng pentamer (penta = 5) hoc hexamer (hexa = 6). Khi lng phn t ca n kh ln, xp x 900 kDa. Chui J thng thy gn vi nhiu pentamer, trong khi cc hexamer li khng cha chui J do cu trc khng gian khng ph hp. Do mi monomer c hai v tr gn khng nguyn, mt pentamer IgM c 10 v tr gn khng nguyn, tuy vy n khng th gn cng lc 10 antigen v chng cn tr ln nhau. V l mt phn t ln, IgM khng c kh nng xuyn thm, n ch tn ti vi lng rt nh trong dch k. IgM ch yu trong huyt tng, chui J rt cn cho dng xut tit. Nh tnh cht polymer, IgM rt "hu" khng nguyn v rt hiu qu trong vic hot ha b th. N cn c gi l cc "khng th t nhin" v lu hnh trong mu ngay c khi khng c bng chng v s tip xc vi khng nguyn. cc t bo dng mm, segment gene m ha vng hng nh ca chui nng c gii m trc cc segment khc. Do , IgM l immunoglobulin u tin c sn xut bi t bo B trng thnh. IgE IgE l loi immunoglobulin monomer trong carbonhydrate chim t l kh ln. Khi lng phn t ca IgE l 190 kDa. IgE c trn mng bo tng ca bch cu i kim v t bo mast m lin kt. IgE gi mt vai tr trong phn ng qu mn cp cng nh trong c
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ch min dch chng k sinh trng. Khng th loi IgE cng c trong cc dch tit, khng hot ha b th v l loi immunoglobulin d b hy bi nhit. IgD IgD l loi immunoglobulin monomer chim cha y 1% trn mng t bo lympho B. Chc nng ca IgD cha c hiu bit y , n thng biu hin ng thi vi IgM v c xem nh mt ch du (marker) ca t bo B trng thnh nhng cha tip xc khng nguyn. C l n tham gia vo c ch bit ha ca t bo B thnh tng bo v t bo B ghi nh. 2.2. C ch tc ng ca khng nguyn- khng th Trong mt p ng min dch, khng th c 3 chc nng chnh: gn vi khng nguyn, kch hot h thng b th v huy ng cc t bo min dch. Cc immunoglobulin c kh nng nhn din v gn mt cch c hiu vi 1 khng nguyn tng ng nh cc domain bin thin. Mt th d miu t li ch ca khng th l trong phn ng chng c t vi khun. Khng th gn vi v qua trung ha c t, ngn nga s bm dnh ca cc c t trn ln cc th th t bo. Nh vy, cc t bo c th trnh c cc ri lon do cc c t gy ra (hnh 12.4(a) v 12.4(b). Tng t nh vy, nhiu virus v vi khun ch gy bnh khi bm c vo cc t bo c th. Vi khun s dng cc phn t bm dnh l adhesine, cn virus s hu cc protein c nh trn lp v ngoi. Cc khng th khng-adhesine v khng-proteine capside virus s ngn chn cc vi sinh vt ny gn vo cc t bo ch ca chng.
(a) (b) Hnh 12.4 (a) : Cc c t ca vi khun bn cnh mt t bo c th. Hnh 12. 4 (b): Cc c t trn b trung ha bi khng th. Hot ha b th Mt trong nhng c ch bo v c th ca khng th l vic hot ha dng thc b th. B th l tp hp cc protein huyt tng khi c hot ha s tiu dit cc vi khun xm hi bng cch (1) c cc l thng trn vi khun, (2) to iu kin cho hin tng thc bo, (3) thanh lc cc phc hp min dch v (4) phng thch cc phn t ha hng ng. Hot ha cc t bo min dch Sau khi gn vo khng nguyn u bin thin (Fab), khng th c th lin kt vi cc t bo min dch u hng nh (Fc). Nhng tng tc ny c tm quan trng c bit trong p ng min dch. Nh vy, cc khng th gn vi mt vi khun c th lin kt vi mt
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i thc bo v khi ng hin tng thc bo. Cc t bo lympho NK (Natural Killer) c th thc hin chc nng c t bo v ly gii cc vi khun b opsonine ha bi cc khng th. Khng th n dng v a dng Khng th n dng Cc khng th n dng ch nhn bit mt epitope trn mt khng nguyn cho sn (hnh 12.5). Theo nh ngha, tt c cc khng th n dng cng mt dng th ging ht nhau v c sn xut bi cng mt dng tng bo. Khng th n dng c s dng rng ri trong sinh hc v y hc, chng va l phng tin chn on, va l cng c iu tr. Th d, chng c ng dng trong mt phng php pht hin c thai c s dng ph bin hin nay. Trc y, vic sn xut khng th n dng in vitro rt kh khn do i sng ngn ngi ca cc tng bo. Khng th ch thu c in vivo bng cch tim mt khng nguyn c th vo mt ng vt ri chit ly khng th trong mu. Phng php ny rt tn km nhng ch thu c lng khng th rt t, khng thun nht v b nhim. Mt tin b to ln t c cui nhng nm 1970 bi Cesar Milstein v Georges Khler vi k thut hybridoma (t bo lai gia 1 lympho B c kh nng sn xut khng th vi 1 t bo ung th c i sng kh di). Mt tc nhn gy bnh (vi khun, virus, v.v.) c h min dch nhn din nh l cc khng nguyn. Thng thng, mt khng nguyn c th gm nhiu epitope khc nhau.
Hnh 12.5: Khng th n dng, lin kt vi mt epitope c hiu (tri). Hnh 12.6: Cc khng th a dng, mi khng th lin kt vi mt epitope khc nhau (phi). Khng th a dng Cc khng th a dng l mt tp hp cc khng th c hiu vi cc epitope khc nhau trn mt khng nguyn cho trc (xem Hnh 12.6). Trong p ng min dch, c th tng hp nhiu khng th tng ng vi cc epitope ca cng mt khng nguyn: p ng nh vy gi l a dng. 2.3. Cc hnh thc tc ng ca khng th Rt a dng, c th l: Trung ho c t do lng kt.
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Dnh kt cc vi khun hay cc t bo khc. Lm tan cc vi khun khi c mt ca b th trong huyt thanh bnh thng. Dn d v giao np cc vi khun cho qu trnh thc bo. 2.4. Vai tr ca khng th Trong mt p ng min dch, khng th c 3 chc nng chnh: gn vi khng nguyn, kch hot h thng b th v huy ng cc t bo min dch. Khng th xanthones, thn dc: Nc mng ct thng phm ny l nc gii kht c tc dng tng cng h thng min nhim. Tuy nhin, i vi ngi VN t lu tng s dng v mng ct : - Chng cc bnh truyn nhim - Ci tin ln da - Ngn nga cc bin chng ca i mt - Gip ming l mau lnh - Gip ngn nga sn thn - Ci tin h thng tiu ha
3. B th
B th (complement) l mt nhm protein huyt thanh c c im ho hc v min dch hc khc nhau, c kh nng phn ng vi nhau, vi khng th v vi mng sinh cht ca t bo. H thng b th bao gm 9 protein c k hiu l C (t C1 n C9), cc yu t B, D v propecdin. Cc b th mun hot ng phi c hot ho. S hot ho ny theo kiu phn ng dy chuyn. Mt b th sau khi c hot ho s kch thch hot ho b th tip theo. C hai cch hot ho b th. Cch th nht pht hin sm hn gi l hot ho theo con ng c in. Cch th hai pht hin sau gi l hot ho theo con ng nhnh hay con ng propecdin. Hai con ng ny ch khc nhau giai on u, phn cht li ging nhau. Hot ho theo con ng c in cn c s tham gia ca cc b th C1, C4, C2 v khng th. C th chia ra lm su bc. 1. To phc h khng nguyn- khng th hot ho C1.C1 gm 3 protein hp thnh. C1q, C1r v C1s. Khi C1 gn vo phn ca FC ca 2IgG (thuc loi IgG1, IgG2, IgG3 thuc IgM) nm k nhau, th C1q c hot ho. Tiu phn ny sau khi c hot ho s kch thch C1r, C1r c hot ho s kch thch C1S, C1S hot ho s tham gia hot ho C4 v C2. 2. C1S hot ho s xc tc tch i C4 thnh C4a v C4b. Mnh C4b s gn vo mng nguyn sinh cht ca t bo v vo C2 to thnh C4bC2, C1S hot ho, tip tc tch i C2 thnh C2a v C2b. C4b kt hp vi C2a thnh C4b2a, chnh l convertase C3 (enzym b C3).
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3. Di tc dng ca C4b2a, C3 b tch thnh 2 mnh l C3a v C3b. C3b gn vi C4b2a thnh C4b2a3b chnh l convertase C5 (enzym b C5). 4. Di tc dng ca C4b2a3b, C5 b tch thnh 2 mnh l C5a v C5b. 5. C5b gn vo t bo ch v vo C6 to phc hp C5b6 sau gn vi C7 to thnh C5b67. Phc hp ny bm vo lp lipid kp ca mng sinh cht. 6. C5b67 bm vo C8 sau bm tip vo C9 to thnh phc hp tn cng mng C5b6789 khoan l thng, lm thot ni cht v git cht t bo. Hot ho theo con ng nhnh khng cn c s tham gia ca khng th, nhng cn n yu t B, D v propecdin thay cho cc b th C1, C2, C4 dn n to thnh enzym hot ho C3.
Hnh 12.7. Hot ho v iu ho theo con ng c in C hai con ng u to thnh convertase C3. C4b2a( con ng c in) v C3bBb (con ng nhnh). Cc enzym ny gn vi C3b to thnh covertase C5. C4b2a3b (con ng c in) C3bBb3b (con ng nhnh). Yu t B gn vi C3b to phc hp C3bB. Khi gn vi C3b, yu t B l v tr chu s tc ng ca yu t D v b tch ra thnh Ba v Bb. Mnh Bb c gi li to thnh phc hp C3bBb, chnh l convertase C3 dng b C3. C3bBb gn thm C3b to thnh C3bBb3b chnh l convertase C5. T y s hot ho b th s din ra ging nh con ng c in. S hot ho b th c iu ho bi cc cht c ch. Cht bt hot C1 l C1-INH, cn cht bt hot C3b l yu t I, yu t H cnh tranh vi yu t B gn vi C3b ngn cn s to thnh C3bBb.
Hnh 12.8. S hnh thnh phc h tn cng mng
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S hot ho b th dn n nhiu hu qu sinh hc, trc ht l lm tan t bo nh phc h tn cng mng (C5b6789). C3b va gn vi th th ca vi khun, virus va gn vi i thc bo, do lm tng kh nng thc bo. C3b bao ph quanh t bo ch s lm tng kh nng tiu dit t bo ch ca t bo K trong hiu ng ADCC (gy c t bo ph thuc khng th). (Phn Fab ca khng th c hiu bm vo t bo ch, cn phn FC bm vo t bo K, gip cho t bo K dit t bo ch). C3a v C5a l c t gy phn v, chng tham gia vo s gii phng histamin khi t bo Mast. C3a v C5a cng l cht ho ng ng, chng li ko bch cu trung tnh vo ni tp trung khng nguyn l lm nhim v thc bo.
III. VACCINE V HUYT THANH MIN DCH

1. Vaccine
1.1. nh ngha Vaccine l ch phm c tnh khng nguyn dng to min dch c hiu ch ng, nhm tng sc khng ca c th i vi mt (s) tc nhn gy bnh c th. Thut ng vaccine xut pht t vaccinia, loi virus gy bnh u b nhng khi em chng cho ngi li gip nga c bnh u ma (ting Latinh vacca ngha l "con b ci"). Vic dng vaccine phng bnh gi chung l chng nga hay tim phng hoc tim chng, mc d vaccine khng nhng c cy (chng), tim m cn c th c a vo c th qua ng ming. 1.2. Cc loi vaccine a. Cc vaccine cht Vaccine t, thng hn, ho g (VK) Cm, di, bi lit (virus) Cc vaccine lao (BCG), cm, si, bi lit Ch : Hiu lc cao vaccine cht Nhc: Mt s khi tim vo c th vn xut hin nguy c l mt s t bo c th tr li dng c ban u. Bch hu un vn c t do chng tit ra l tc nhn gy bnh, khi dng ho cht ph hu c tnh nhng vn gi nguyn tnh khng nguyn ca c t
b. Cc vaccine sng
c. Cc vaccine gii c t -
1.3. Nguyn l s dng vaccine S dng vaccine l to min dch ch ng nhn to.
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Tim chng trn phm vi hp l, t t l cao. Tim chng ng i tng. Bt u tim chng ng lc; bo m ng khong cch gia cc ln tim chng; tim chng nhc li ng thi gian. Tim chng ng ng v ng liu lng. Nm vng phng php phng v x tr cc phn ng khng mong mun do tim chng. Bo qun vaccine ng qui nh.
1.4. Mt s loi vaccine mi ang nghin cu a. S dng cc ph gia (adjuvant) mi, nhm gy ra loi p ng min dch mong mun. Th d, cht nhm phosphate v cc oligonucleotide cha CpG demethyl ha a vo vaccine khin p ng min dch pht trin theo hng dch th (to khng th) thay v t bo. b. Vaccine khm: s dng mt sinh th quen bit hn ch hin tng "phn tc dng", th d dng virus vaccinia mang mt s yu t ca virus vim gan B hay virus di. c. Vaccine polypeptide: tng cng tnh sinh min dch nh lin kt tt hn vi cc phn t MHC: peptide nhn to 1/2 ging virus, 1/2 kia gn MHC; on peptide m phng 1 quyt nh khng nguyn (epitope). d. Anti-idiotype: idiotype l cu trc khng gian ca khng th ti v tr gn khng nguyn, c hiu vi khng nguyn tng ng. Anti-idiotype l cc khng th c hiu i vi idiotype, do anti-idiotype xt v mt c hiu li tng t vi khng nguyn. Vy, thay v dng khng nguyn X lm vaccine, ngi ta dng idiotype anti-anti-X. g.Vaccine DNA: DNA ca tc nhn gy bnh s c biu hin bi t bo ngi c chng nga. Li th ca DNA l r, bn, d sn xut ra s lng ln nn thch hp cho nhng chng trnh tim chng rng ri. Ngoi ra, vaccine DNA cn gip nh hng p ng min dch: tc nhn gy bnh ngoi bo c trnh din qua MHC loi II, dn n p ng CD4 (p ng min dch dch th). Khi khng nguyn ca tc nhn c chnh c th ngi biu hin, n s c trnh din qua MHC loi I, lc ny p ng min dch t bo qua CD8 c kch thch. Tuy nhin phng php ny l con dao hai li bi l t bo mang DNA l c nguy c b nhn din l "khng ta", sinh ra bnh t min. h. S dng vector ti t hp dng cc vi khun thun tnh hoc cc t bo trnh din khng nguyn nh t bo tua c chuyn gen biu hin khng nguyn mong mun.
2. Huyt thanh
2.1. Nguyn l s dng huyt thanh S dng huyt thanh l a vo c th khng th c ngun gc t ngi hoc ng vt, gip cho c th c ngay khng th c hiu chng li tc nhn gy bnh. 2.2. Nguyn tc s dng Cc nguyn tc c bn phi thc hin khi s dng huyt thanh l:
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ng i tng ng liu lng ng ng phng phn ng Phi hp s dng vaccine 2.3. ng a huyt thanh vo c th Huyt thanh thng c a vo c th bng ng tim bp. i vi nhng loi huyt thanh c tinh ch t tiu chun cao, c th tim tnh mch nhng cng rt nn hn ch. Tuyt i khng tim tnh mch nhng huyt thanh c ngun gc t ng vt (d c tinh ch!) hoc huyt thanh ngi cha t tinh ch cao. Ngi t nn mng cho vic s dng huyt thanh cha bnh l E. Behring (1854-1917) bng cc cng trnh nghin cu v vic dng khng c t cha bnh bch hu v un vn vo nm 1890. Vi nhng cng trnh nghin cu ny, ng c nhn gii thng Nobel vo nm 1901.
IV. MIN DCH

1. nh ngha:
Min dch (MD-immunity) l trng thi bo v c bit ca c th chng li cc tc nhn gy bnh (vi sinh vt v cc c t ca chng, cc phn t l...) khi chng xm nhp vo c th.
2. C ch bo v khng c hiu
L min dch t nhin mang tnh cht bm sinh, bao gm cc tuyn phng th t ngoi vo trong ngn chn khng cho vi sinh vt (VSV) xm nhp vo c th. - Hng ro vt l. Da v nim mc ngn cch ni mi ca c th vi ngoi mi xung quanh. Trn mt biu b cha keratin khng tan trong nc, khng b VSV phn gii, do khng cho VSV thm qua. Pha ngoi, biu b ch yu gm t bo cht nn virus khng nhn ln c. Nim mc bao ph b mt trong c th nh ng h hp, tiu ho v sinh dc. Cht nhy ca nim mc l ci by bt gi VSV, h thng nhu mao chuyn ng ht VSV ra bn ngoi qua ng mi hoc ming. Lng mi, phn x ho, ht hi, nn cng gip ngn chn VSV xm nhp vo c th. * Dch c th nh nc mt, nc bt, nc tiu...cng gip ra tri VSV khi chng bm trn b mt cc m, y ht nh ta tm ra lm tri bi bm. - Hng ro VSV bao gm cc vi khun bao ph b mt hoc khu tr bn trong c th, to nn khu h VSV bnh thng. y l mt khu h VSV phc tp, c s lng gp 9 ln tng s t bo ca c th. Chng chim trc cc v tr m VSV gy bnh n, cnh tranh thc n v tit ra cht ho hc tiu dit VSV gy bnh. Ngoi ra nhiu vi khun sng trong ng tiu ho cn tit ra biotin, riboflavin v cc vitamin khc cung cp cho c th. Khi ph v s cn bng trong khu h, mt s s c th tr thnh VSV gy bnh c hi.
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- Hng ro ho hc. trong dch tit ca cc m trong mu, trong dch lympho u cha cc nhn t c ch hoc tiu dit VSV gy bnh. * pH acid c ch s sinh trng ca VSV. VSV phn gii lipid thnh acid bo lm tng acid ca da. Vi khun lactic ln men glycogen to acid lactic trong m o. Dch v do tuyn trong d dy tit ra cha acid clohydric c pH 1-2. Sn phm ln men ca cc vi khun thng tr trong ng tiu ho l acid lactic v axetic. Ngoi ra VSV cn tit ra bacterioxin l cht dit khun mnh. * Lysozym l enzym c trong nc mt, nc mi, nc bt, dy tai, m hi, sa, dch m o...v.v.., c kh nng dit vi khun Gram dng nh c ch phn gii thnh t bo, do c coi l mt loi cht khng sinh. Protein lin kt vi st nh lactoferrin c trong tinh dch, nc mt, sa m, mt, nc mi, dch nhy rut...v.v.., transferrin c trong gian bao cc m, trong huyt thanh lm gim trng lng st t do gy tnh trng thiu st cn thit cho VSV sinh trng, do ngn cn c nhim trng. Inteferon (IFN) l glycoprotein do cc t bo m nhim virus hoc vi khun (IFN v IFN-) hoc do cc t bo lympho T sinh ra (IFN-). IFN c ch s nhn ln ca virus trong t bo bng cch hnh thnh protein cm ng ngn cn qu trnh dch m v phn hu RNA thng tin ca virus. Ngoi vai tr bo v, IFN cn lm nhim v iu ho mng li bo v chng nhim trng v s nhn ln ca cc t bo ung th. B th (Complement) l glycoprotein c kh nng lm tan t bo v tng cng hin tng thc bo. - Hin tng thc bo. Cc bch cu tham gia vo c ch bo v khng c hiu bao gm. Bch cu trung tnh cha cc ht khng bt mu thuc nhum, c tnh ho ng ng bt gi v tiu dit cc vt l, k c VSV v cc mnh t bo phn hu, do vy ng vai tr quan trng trong chng nhim trng. Bch cu a kim cha cc ht bt mu xanh methylen, cha cc hot cht nhu histamin, heparin, serotonin. Bch cu a acid cha cc ht bt mu eosin, cha enzym phn gii histamin v heparin, do vy iu ho hot ng ca bch cu kim. Bch cu ny c kh nng gy c cc u trng giun. i thc bo ng vai tr quan trng trong chng nhim trng, nht l virus. i thc bo thu tm VSV nh gi tc, to khng bo (phagosom). Cc khng bo dung hp vi lysosom to phagolysosom. Cc VSV trong phagolysosom b tiu dit do nhiu nguyn nhn. - Cc enzym ph thuc oxy chuyn ho oxy phn t thnh peoxit gy c cho VSV. - S thay i kiu h hp, to acid lactic lm gim pH, pH thp lm tng hot tnh nhiu enzym trong lysosom. - C ch thc bo khng ph thuc oxy bao gm hot ng ca cc enzym phn gii ca lysosom nh lyzozim, photpholipase, ribonuclease, deoxyribonuclease..v.v.. St l c ch bo v t nhin, lm tng phn ng enzym phn hu VSV, lm tng hot ng ca IFN v lm gim nng st t do trong mu, do gp phn tiu dit VSV.
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Vim khng c hiu c tc dng khu tr VSV vo mt ni khng cho chng lan rng thm, 4 tnh cht ca vim l. , au, sng, nng l do gin mch, tng dng mu, lm thot bradykinin v prostaglandin, bch cu trung tnh, i thc bo vo vim.
3. Cht sinh min dch (immunogen)

L nhng cht khi a vo c th iu kin thch hp c kh nng gy mt p ng min dch (UMD), cn khng nguyn (KN-antigen) l nhng cht c kh nng lin kt vi khng th hoc th th c hiu ca t bo limpho T (TCR). Tt c cc cht sinh min dch u l khng nguyn, nhng khng phi tt c khng nguyn u l cht sinh min dch. V d. Hapten l khng nguyn khng trn vn, c th gn vi khng th nhng khng kch thch to khng th. Ba iu kin cn ca mt cht sinh min dch l. - Tnh l. C th khng p ng bo v vi khng nguyn bn thn, do vy cht cng l vi c th, kh nng gy p ng min dch cng cao. - Trng lng phn t ln. Khng nguyn thng c trng lng phn t trn 10.000 dalton. Nu thp hn, kh nng sinh min dch yu hoc khng c v kh b i thc bo bt gi v x l. - Cu trc phn t phc tp. nhiu cht c trng lng phn t ln nh polylisin, polisaccharid khng gy hoc gy p ng min dch yu v cu trc n gin do lp i lp li, trong khi hapten c trng lng phn t thp nhng gn vi protein th li tr thnh cht gy p ng min dch.
4. Tnh c hiu ca khng nguyn (KN)

Khng phi ton b khng nguyn tham gia vo kch thch h thng min dch m ch c mt phn nht nh ca khng nguyn gi l quyt nh khng nguyn hay epitop, mi lin kt vi khng th hoc t bo lympho. Phn tng ng vi quyt nh khng nguyn nm trn mi khng th gi l v tr kt hp khng nguyn hay paratop cn phn tng ng nm trn mt t bo lympho l th th, v d th t bo T (TCR T-cellreceptor). S lin kt gia epitop vi paratop mang tnh c hiu cao, tng ng nh gia enzym v c cht hay gia kho vi cha.
5. Cc c quan v t bo tham gia vo h thng min dch

Nhiu c quan v t bo nm ri rc khp c th hp tc vi nhau nhn din v phn ng vi khng nguyn theo nhiu cch dn n p ng min dch cui cng. 5.1. cc c quan lympho L ni hun luyn v tng tr cc t bo lympho, bao gm c quan lympho trung tm v c quan lympho ngoi vi. a. C quan lympho trung tm gm tuyn c v ti fabricius Cc t bo gc hay cn gi l t bo ngun i t tu xng vo tuyn c. Ti phn v ca tuyn c, tin t bo T c phn chia nhiu ln v bit ho thnh t bo T chn vi cc th th khc nhau trn b mt. Cc gen kim tra s hnh thnh cc th th ny xut hin
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do ti t hp DNAca t bo T non khi cn tuyn c. Phn ln cc t bo T c th th nhn din c khng nguyn bn thn s b loi tr v cht. Cc t bo cn li di c vo phn tu ca tuyn c tip tc bit ho to cc dng t bo T phn lp, sau cc t bo ny theo mu ri tuyn c vo c quan lympho ngoi vi. Ti Fabricus l c quan lympho trung tm ch c gia cm, nm gn hu mn v l ni bit ho t bo B. ng vt c v khng c ti Fabricus nn s bit ho t bo B xy ra ngay trong tu xng v cc c quan tng ng, nh c quan lympho ca h tiu ho. Cc t bo B chn ri ti n c quan lympho ngoi vi. Khi tip xc vi khng nguyn s c kch thch, bit ho tr thnh t bo plasma (tng bo) sn xut khng th. V th t bo plasma c coi l nh my sn xut khng th. b. Cc c quan lympho ngoi vi Lch l c quan lympho ngoi vi ln nht, l ni by khng nguyn vo bng ng tnh mch v l c quan chnh to khng th. Trong lch c vng cha t bo T v vng cha t bo B. Trung tm mm nm trong nang lympho vng t bo B. Trong trung tm mm cha y t bo B v t bo plasma, mt s i thc bo v t bo tua. Hch lympho nm ri rc khp c th v hot ng nh mt h thng lc. Khng nguyn di chuyn chm theo cc mch hp v gp khc tng s tip xc vi i thc bo v t bo lympho. Phn v gm vng v nng c nhiu nang cha t bo B. Khi c khng nguyn kch thch, nang s ni rng ra to trung tm mm, cha ch yu t bo B chun b phn chia. Vng v su cha ch yu l t bo T v mt s i thc bo. Phn ty c nhiu xoang cha dch lympho. Cc t bo plasma i t vng v sang vng tu. Khi c khng nguyn xm nhp, i thc bo bt gi, x l, hp tc vi cc t bo B v T sn xut khng th. 5.2. Cc t bo tham gia v p ng min dch. T ngun gc chung l t bo tu xng s c bit ho to thnh cc dng t bo khc nhau. (Hnh 12.9) - Dng to mu bit ho thnh cc t bo dng tu bao gm t bo mono (tin thn ca i thc bo) v cc t bo a nhn (bch cu trung tnh, bch cu kim v bch cu acid). Dng hng cu to t bo hng cu v dng t bo khng l to tiu cu. - Dng lympho i vo cc c quan trung tm. Nu vo ti Fabricius, s bit ho thnh t bo B (t ch Bursa Fabricius) khi c khng nguyn kch thch t bo B s bit ho thnh t bo plasma sn xut khng th v t bo B nh p ng vi khng nguyn vo ln hai. Nu t bo ngun i vo tuyn c s c bit ho thnh cc t bo lympho T (t ch Thymus-tuyn c). Cc t bo T c phn bit bi cc th th b mt thnh cc dng t bo T4 v T8. Dng t bo T4 bao gm. - Lympho T h tr (TH) c nhim v kch thch t bo B bit ho thnh t bo plasma sn xut khng th.
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Hnh 12.9. Ngun gc cc t bo to mu v t bo tham gia vo h min dch - Lympho T cm ng (Ti) tit ra intlkin-2 kch thch s bit ho v hot ng ca cc t bo T khc. - Lympho T gy qu mm mun (TDTH) tit lymphokin hot ho i thc bo v cc bch cu khc gy qu mm mun. - Lympho T iu ho ngc (TFR) c tc dng hot ho lympho T c ch. Dng t bo T8 bao gm. - Lympho T c (TC) tn cng trc tip cc t bo c khng nguyn l (t bo ung th, t bo nhim virus). - Lympho T c ch (TS) iu ho p ng min dch bng cch c ch s hot ng ca cc loi lympho bo khc. i thc bo ng vai tr nh l t bo khi mo cho mt p ng min dch. Trc ht chng lm nhim v ch bin khng nguyn bng cch bt gi, nut v tit enzym phn gii bc l quyt nh khng nguyn, sau y khng nguyn ra b mt tip cn vi t bo B v t bo T. Nh ng tc ny m i thc bo c gi l t bo trnh din khng nguyn (APC-antigen presenting cell). i thc bo cn cha cc th th dnh cho phn Fc ca IgG v b th C 3, do lm tng kh nng thc bo.
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6. Min dch bnh l

6.1. Qu mn L nhng tn thng bnh l xy ra do mt p ng min dch qu mc v khng hp l, gy hu hoi m. Qu mn khng biu hin ngay sau khi tip xc vi khng nguyn ln u, m ch nhng ln tip xc sau. Gell v Combs chia qu mn ra l 4 loi. * Qu mn tip I hay qu mn tc th c c trng bi cc phn ng d ng, xy ra nhanh (<30pht) ngay sau khi tip xc vi khng nguyn vo ln hai, nguyn do l IgE c hnh thnh gn vo th th FC ca t bo mast, phn Fab ca hai IgE nm k nhau gn cho vi khng nguyn, dn n lm thay i cu trc khng gian ca th th, gy bin i ti ch mng, lm thot cc bng ri t bng gii phng cc cht trung gian hot mch nh histamin (gy co tht c trn), serotonin (tng tnh thm thnh mch), heparin (km hm ng mch, ECF-A-yu t ho hng ng bch cu trung tnh). S hot ho t bo Mast cng dn n vic tng hp cc cht mi nh phn ng chm (SRS-A lm tng tnh thm thnh mch) v yu t hot ho tiu cu (PAF, gy ngng kt tiu cu v gii phng histamin). Qu mn tp I c th biu hin ton thn nh sc phn v, sc do d ng vi thuc, sc do dng huyt thanh iu tr hoc bnh cht trong ni (khng th chng protein c trong sa b). Qu mn loi ny cng c th biu hin cc b nh hen ph qun v vim mi d ng). * Qu mn tp II hay qu mn gy c t bo xy ra khi khng th gn trc tip vi khng nguyn trn b mt t bo cng vi s tham gia ca b th v cc t bo hiu ng (t bo K, tiu cu, bch cu trung tnh, bch cu n nhn hay i thc bo) dn ti lm tan t bo ch, chng hn t bo ch gn vi khng th c hiu, hot ho b th to phc hp C5b-9 tn cng mng t bo ch, ng thi mnh C3b c th bm quanh t bo ch v bm vo i thc bo thc hin opsonin ho, gip i thc bo dit t bo ch. Qu mn tp II th hin rt r trong cc trng hp sau. Khi truyn mu khng ph hp, chng hn truyn mu nhm A cho ngi c nhm mu B th s lm tan mu ca khng th (IgM) ca nhm A s khng li khng nguyn B ngi nhn. Phn ng khng nguyn-khng th s hot ho b th gy hin tng tan hng cu ca ngi nhn mu, gy tn thng thn do tc nghn bi mt lng ln mng hng cu v gy c do gii phng phc hp hem (C34H23O4N4Fe). Bng 12.10. S nhm mu Nhm Khng nguyn AB A B O A&B A B Khng c Khng th Khng c khng th Khng th khng B Khng th khng A Nhm c th thn AB, O, A, B (nhn ton nng) A, O B, O
Khng th khng A, O (cho ton nng) khng B
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Nhm mu Rh. khi b mang mu Rh+ v m mang mu Rh- th 50% a tr cho i mang Rh+. Nu thai nhi mang Rh+ th hng cu c th truyn sang m, kch thch to khng th khng Rh. Khng th ny c th truyn cho con qua nhau thai gy tan hng cu ca thai nhi. trnh tai ho cho ln c thai sau, ngi ta phi tim cho m khng th khng Rh (RhoGAM) loi b khng nguyn Rh c th m. Bnh ban xut huyt xy ra khi phn t thuc (hapten) gn xung quanh tiu cu, lc chng tr thnh khng nguyn mnh. Khng th chng phc hp ny gy hu tiu cu khi c mt b th. V tiu cu cn cho s ng mu nn nu thiu s gy xut huyt trn da. * Qu mn tp III hay phc hp min dch khc vi qun mu tp II ch khng th chng li vi khng nguyn ho tan phn b rng ri trong mu. Khi tim khng nguyn vo c th s kt hp vi khng th kt ta c sn to thnh phc hp khng nguyn- khng th ri lng ng vo gia cc t bo ni m v mng y mao mch. Hin tng ny ko theo s hot ho dng b th to C3a v C5a (cc cht gy phn v) lm tng tnh thm thnh mch (gay ph n). Cc thnh phn khc l cc cht ho ng ng, thu ht bch cu trung tnh. Cng vi tiu cu, cc t bo ny tp trung gy ghn mao mch. Bch cu trung tnh hot ho s tit enzym protease, colagenase v cc cht hot mch gy vim lot. Kt qu l mao mch t, xut huyt dn n hoi t cc b. Hin tng ny gi l phn ng Arthus. Bnh vim cu thn l mt v d ca qu mn tp III. Phc hp khng nguyn-khng th ng li trong gian bo ni m v mng y vch mao mch gy vim. Khng nguyn c th l vi khun (sau nhim lin cu), k sinh (vin thn sau st rt c tnh ) protein l (vim thn trong bnh huyt thanh). Bnh huyt thanh. vic s dng khng huyt thanh trong iu tr mang li kt qu ln (v nh virrut di), song mt s trng hp c th gy tai bin. Nguyn nhn l do phc hp min dch lng ng trong thnh mch mu. Bnh phc hp min dch lin quan n nhim trng (Rheumatic fever). khi b vim hng do Streptococcus gy ra, khng nguyn thnh t bo cha protein M kch thch to khng th. Ti mng tim, thn, khp cng c protein ny, do vy khng th khng vi khun gy phn ng cho vi cc thnh phn ca c th gy vim (mng tim, thn, khp). V d. vim cu thn * Qu mn tp IV hay qu mn mun l tn thng do s tng tc gia khng nguyn vi cc lympho T mn cm vi cc khng nguyn gy ra. Phn ng xy ra mun, sau 24 gi hoc hn. Cc thng tn c th do t bo TC ph hu t bo ch c cc
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khng nguyn c hiu bm trn b mt hoc do tc ng ca lymphokin do lympho T tit ra, c tc dng lm cho i thc bo tp hp qu nhiu, nhp li thnh t bo khng l (u ht). Qu mn do tip xc l mt v d. Mt s cht nh niken, cromat, cc ho cht trong cao su, ho, m phm...c trong lng phn t (<1kDa) nn l hapten. Khi thm qua biu b, gp protein s tr thnh cht cng hp hapten protein thi (khng nguyn) kch thch lympho T, c bit l TC. Mi tng tc gia lympho T ny vi hapten bm trn da gy phn ng vim da. 6.2. Bnh t min Bnh thng c th khng sinh p ng min dch chng li khng nguyn bn thn. Khi tnh t dung np b mt v c th khng cn kh nng phn bit khng nguyn bn thn vi khng nguyn l th s gy p ng min dch chng li khng nguyn bn thn v dn n bnh t min. Bnh t min c th thuc qu mn tp II, III hoc IV. Sau y l mt s v d. + Bnh Grave (Tp II) l do khng th gn vo th th ca t bo tuyn gip dnh cho hocmon kch thch tuyn gip. Hocmon ny do tuyn yn sinh ra. Kt qu l tuyn gip b kch thch sn ra mt lng ngy cng ln hocmon tuyn gip dn n bu c do tuyn gip phng to. + Bnh nhc c (Tp II) lm cho c th suy yu. y l do khng th gn vo th th dnh cho axetylcholin ti im ni xung thn kinh c. Do khng tip nhn xung thn kinh do phn t axetylcholin, c s b yu, h hp dng, dn n t vong. + Luput ban h thng (SLE) (tp III) l bnh to trn vng ban hnh bm hay mt si. Nguyn nhn do to thnh khng th chng DNA dng xon kp hoc si n (gi chung l khng th khng nhn). Phc hp min dch lng ng cu thn, hot ho b th, thu ht bch cu ht gy vim cu thn. + Bnh tiu ng ph thuc insulin (Tp IV) do khng th c hiu tiu dit t bo tuyn tu sn sinh insulin. Vic thiu insulin lm cho lng ng trong mu tng dn n m lo v hoi th. Suy gim min dch. Suy gim min dch l s p ng min dch khng y . Nguyn nhn c th do s khim khuyt h thng min dch mang tnh bm sinh, do sai st hoc thiu cc gen ca h thng min dch mang tnh bm sinh, do sai st hoc thiu cc gen ca h thng min dch nhn t cha m hoc c th mc sau khi dng thuc, b ung th hoc sau khi nhim trng. V d. bnh Hodgkin (ung th) lm gim p ng min dch qua trung gian t bo hoc nhiu loi virus git cc t bo ca h thng min dch (HIV). Hi chng suy gim min dch mc phi (AIDS) Virus HIV l virus Retro mang genom RNA mt si v enzym phin m ngc. Bao bc capsit l v ngoi liporotein. Trn mt v ngoi cha cc gai glycoprotein vi trng lng phn t 120 kDa nn gi tt l gp 120. Cc gai ny gn vo th th CD4 nm trn mt t bo TCD4 v i thc bo l t bo ch chnh ca HIV.
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Sau khi xm nhp vo t bo, virus ci b ngoi nh enzym tiu ho ca t bo sau si RNA virus c sao thnh DNA n nh enzym sao m ngc. Si DNA mi tng hp c dng lm khun tng hp si DNAmi theo c ch b sung. Hai si xon vo nhau to si DNAxon kp. DNA kp ny gn xen vo nhim sc th ca t bo ch nh enzym integrase v nm di dng porvirus. Mt bn sao DNA c th nm im trong t bo vi thng n vi nm. Gen HIV c biu hin khi c s kch thch ca yu t phin m ca t bo ch gn vo 2 u lp li di ca provirus. S kch thch ny dn n to thnh cc RNA khc nhau, mt s l RNA genom ca virus, mt s l RNA thng tin dng tng hp protein v capsid. V capsid kt hp vi RNA genom to thnh ht virus (virion) chui ra ngoi lp li vng i mi t bo CD4 khc. Ch mt thi gian ngn sau khi nhim c 1-100 t bo cha HIV. Lc u c ch bo v ca c th lm gim tc nhn ln ca virus nhng sau chung vt qua v nhim ngy cng nhiu vo cc t bo T. Da vo s lng t bo T trong mu xc nh mc nhim. Nu lng t bo T4 trong mu xung di 600/, bnh nhn bt u mt p ng min dch qua trung gian t bo v dn n nhim bnh c hi. Khi mi bt u nhim nhiu dng t bo B c hnh thnh v trong huyt thanh cha mt lng ln IgG, IgM v IgA nhng giai on sau ca AIDS, lng khng th gim t ngt v cc khng th khng gp 120 cng gim theo. Liu php thnh cng nht chng HIV l dng Dideoxynucleozit, ch yu l azidotimidin hoc zidovudin. Cc cht ny c ch enzym phin m ngc ca HIV v do cn tr s tng hp DNA ca virus. Cho n nay mi loi thuc cng ch lm gim s tin trin thnh AIDS. Cch tt nht vn l phng chng, ngn s ly lan theo cc con ng quan h tnh dc, tim chch ma tu v truyn mu nhim.
V. CC CHT C HOT TNH SINH HC

1. Enzym
1.1. Khi nim chung Enzym l cht xc tc, chng thc y nhanh cc phn ng m khng tham gia vo thnh phn phm cui cng. Ton b nhng biu hin hot ng sng ca VSV l kt qu tc ng ca cc cht xc tc hu c (enzym) Nm 1926, I.B.Sumner l ngi u tin tch c enzym urease dng tinh th thun khit. l bc m u cho vic nghin cu bn cht ca enzym. 1930, J.H.Northrop thu c dng tinh th cc enzym nh pepsin, trypsin v chemotrypsin. Bn cht protein ca enzym mi c xc nh. c tnh quan trng nht ca enzym l tc dng c hiu ca n, mi loi enzym tc dng ln mt loi c cht nht nh. Tnh c hiu c khi l tuyt i, khi enzym tc ng ln mt c cht (urease), hoc l tng i (nhm) khi enzym tc dng ln mt loi hp cht c nhng nhm nguyn t nht nh (lipase).
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Hot tnh enzym trong t bo c iu chnh nghim ngt, s tng hp sinh hc cc enzym chu s kim tra ca cc gen. T nm 1926 nhiu loi enzym c thu nhn di dng tinh th nh urease, pepxin v tripxin t vi khun. Ngy nay theo thnh phn ha hc, cc enzym c chia thnh 2 nhm ln: -Cc enzym mt thnh phn: enzym ch l phn t protein to nn, enzym 2 thnh phn ngoi protein ra enzym cn c nhm phiprotein, gi l nhm coenzym. Nhm coenzym thng gp l vitamin, ion kim loi, nucleotit, focfirin... Cu trc ca enzym c 2 thnh phn: phn mang enzym v tm khot ng. Tm hot ng l ni thc hin c ch xc tc ca enzym. Cu trc tm hot ng ph hp vi c cht n xc tc. Tm hot ng c chc nng tip xc vi c cht, nh hng xc tc v tin hnh xc tc. Cc enzym thng gn cht vi t bo VSV, nh nhiu loi enzym oxi ha kh v ch ra ngoi khi t bo b ph hy, ta gi l cc endofecmen. Cn cc enzin d c t bo tit ra mi trng xung quanh nh hydrolase gi l exofecmen. Theo c ch tc dng v bn cht ha hc ca cc enzym m ngi ta chia chng thnh 6 lp, mi lp li chia thnh lp ph v di lp ph (xem gio trnh sinh ha). Hin nay cng ngh sinh hc sn xut c cc ch phm enzym. Trong s 50 enzym m th gii ang tin hnh sn xut, nc ta sn xut c 3 ch phm enzym l amylaza, proteaza th v pectinaza dng kt ta ca protein. Hng nghin cu enzym trong tng lai l tp trung vo nhng vn ch yu nh: nghin cu cu trc ha hc ca cc enzym quan trng v thng dng, tim hiu nhng vn c bn v c ch tc ng ca mt s enzym, tng hp nh hng cc enzym trong t bo VSV. Hin nay enzym c s dng rng ri trong cng nghip thc phm, dc phm, nng nghip v y hc. Nh s gia tng s dng enzym m qui m sn xut c m rng, cc loi ch phm a dng phong ph hn, cht lng ch phm ngy cng c ci tin. 1.2. Cu to ha hc ca enzym. -Bn cht protein ca enzym -Trung tm hot ng ca enzym 1.3. Tnh cht ca enzym -Xc tc -c hiu (kiu phn ng, c cht, nhm) 1.4. C ch tc dng ca enzym To phc enzym- c cht. 1.5.Cc yu t nh hng n vn tc phn ng enzym -Nng enzym -Nng c cht -Cc cht km hm
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-Cc cht hot ha -Nhit v pH mi trng. 1.6.Cch gi tn v phn loi enzym Da vo tnh c hiu phn ng enzym, cc nh khoa hc chia enzym thnh 6 lp: Oxydoreductase, transpherase, hydrolase, liase, isomerase v ligase. 1.7. Cc phn ng enzym ph bin trong sn xut thc phm -Phn ng thy phn -Phn ng oxygen ha- kh sinh hc.
2. Cht khng sinh

2.1. nh ngha Theo Vin s Iu.A.Ovtchinnikov (1987) th cht khng sinh l cc cht c ngun gc thin nhin v cc sn phm ci bin chng bng con ng ho hc c kh nng c ch s pht trin ca vi sinh vt, virus hay t bo ung th ngay nng thp (10 -3- 10-2 microgam/ml). 2.2. Bn cht ha hc Khng sinh l mt nhm cht rt a dng v mt ha hc, trng lcng phn t bin ng t 150- 5000. Trong mt s cht, phn t ca chng ch cha C v H hoc cha C,H,O v N. .. cho n nay ngi ta mi bit c cc khng sinh u th rn. 2.3. Tc ng ca khng sinh 2.3.1. Hot ph khng sinh: l phm vi hot ng ca khng sinh i vi cc VSV m n tc dng. Cht khng sinh c hot ph rng: c tc dng i vi nhiu loi vi khun khc nhau (Gram m v c vi khun Gram dng).V d: Chloramphenicol, tetracyclin, ampicillin. Cht khng sinh c hot ph hp: ch tc ng chuyn bit v d: Isoniasit, ethambutol, PAS, INH tc dng ln vi khun lao. 2.3.2. Khng sinh : Khng sinh l k thut tm hiu kh nng tc dng ca cht khng sinh i vi tng loi vi khun kim nh. C hai phng php lm khng sinh : - Pha long khng sinh trong mi trng lng vi nhng nng khc nhau. Theo di kh nng pht trin (lm c mi trng) ca tng loi vi khun kim nh. - Phng php dng a khng sinh: cho vi khun pht trin trn thch a. Sau cho khng sinh khuch tn trn b mt a thch, o vng v khun. 2.4. C ch tc dng ca khng sinh 2.4.1. c ch tng hp vch t bo Khc vi t bo ng vt, vi khun c vch t bo. Loi b vch hoc ngn cn s to thnh vch u lm tan t bo vi khun. Vch t bo vi khun cha mucopeptit cn gi l murein hay peptidoglycan. Cc penicilline v cephalosporin tc ng ln cc phn t protein
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chn lc c hiu nn c ch s lin kt ngang cui cng ca cu trc mucopeptit ca vch t bo, vi khun d dng b tan. Cc khng sinh khc nh bacitracin, vancomycin, novobiocin, D-cycloserin cng c ch s to vch ca vi khun bng c ch s hnh thnh mucopeptit ca vch . 2.4.2. c ch chc nng ca mng sinh cht Mngtsinh cht hot ng nh mt mng bn thm. Mng ny thm thu cc cht chn lc v nh vy kim sot c cc thnh phn bn trong ca cc t bo. Nu s ton vn ca mng nguyn tng b ph v th cc thnh phn ni bo thot ra khi t bo v t bo cht. Mng t bo ca mt s vi khun v nm d b ph v bi mt s thuc khng sinh hn t bo ng vt do hot tnh tc dng chn lc ca cc khng sinh ny ln cc nhm cu trc ha hc c bit c mng ca vi khun hoc ca nm m khng c t bo ng vt. Polymycin c tc ng mng sinh cht ca vi khun gram m, polyen c tc ng mng nguyn tng ca nm. 2.4.3. c ch tng hp protein Nhiu thuc khng sinh c ch tng hp protein ca vi khun: 2.4.3.1.Chloramphenicol: chloramphenicol v cc dn cht thuc nhm ny ngn cn s kt hp axit amin vo chui peptit mi sinh n v 50S ca ribosome vi khun qua vic cn tr tc ng ca enzym peptidyl-transferase. 2.4.3.2. Cc tetracyclin: tetracyclin v cc thuc khng sinh nhm ny c ch s tng hp protein ca vi khun bng cch ngn cn s gn lin ca amino acyl-RNA vn chuyn vo n v 30S ca ribosome . 2.4.3.3. Cc macrolid v lincomycin: Cc khng sinh nhm ny ngn cn s tng hp protein bng cnh tranh v tr kt hp ca axit amin ribosome v bng cch phong b phn ng dy chuyn aminoacyl. 2.4.3.4. Cc aminoglycoside: cc thuc khng sinh nhm aminoglycoside c ch tng hp protein bng cch gn vo protein tip nhn trn n v 30S ca ribosome lm c sai thng tin ca RNA thng tin lm hnh thnh cc protein khng c hat tnh, ngoi ra cn lm tch cc ribosome trng thi polymer thnh monomer 2.4.4. Tc dng c ch s hnh thnh acid nucleic 2.4.4.1. Cc sulfonamid v trimethoprim: i vi nhiu vi sinh vt, axit paraaminobenzoic (vit tt l PABA) l mt cht chuyn ha cn thit trong qu trnh tng hp axit folic cn thit tng hp purin v DNA. Sulfonamid do c cu trc tng t nh PABA nn c th i vo phn ng thay cho PABA, hu qu l mt cht tng t nh axit folic nhng khng c hat tnh c to thnh v ngn cn s pht trin ca vi khun. Trimethoprim c ch enzym dihydro folic reductase, enzym ny bin i axit dihydro folic thnh axit tetrahydro folic, mt giai an trong chui phn ng tng hp purin v DNA. S phi hp mt trong cc sulfonamid vi trimethoprim, hai cht tc ng hai khu khc nhau ca mt qu trnh tng hp lm tng r hat tnh ca thuc, cotrimoxazol (sulfamethoxazol v trimethoprim) l ch phm phi hp s dng hiu qu trong iu tr cc bnh nhim trng
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2.4.4.2. Rfampin: Rifampin v cc dn cht kt hp vi RNA polymerase ph thuc DNA v nh th c ch s tng hp RNA vi khun. 2.4.4.3.Cc quinolone: Cac quinolon v cacboxy fluoroquinolon kt hp vo DNA gyrase nn c ch tng hp DNA. 2.5. Cc nhm thuc khng sinh 2.5.1. Sulfonamid : L nhng hp cht thay th nhm amit (SO2NH2) hoc nhm amin (NH2) ca axit sulfanilamic. Sulfonamid c tc dng cn khun i vi vi khun gram m v vi khun gram dng. Chng c ch nhiu vi khun gram dng v vi khun ng rut bao gm E.coli, Klebsiella v Proteus. Cc sulfonamid c s dng rng ri nh sulfaguanidin, sulfadiazim, sulfisoxazol, sulfamethoxazol, sulfadimethoxin.... Tr ngi chnh ca cc thuc nhm sulfamid l vi khun tr nn khng vi thuc nhanh chng, thm vo nhiu sulfamid c th gy phn ng ph nghim trng. Nn hin nay cc thuc nhm sulfamid ch c dng hn ch. Ch phm sulfamid ang c dng rng ri hin nay l cotrimoxazol l phi hp ca sulfamethoxazol v trimethoprim. Thuc ny c dng trong iu tr cc nhim trng ng tiu ha, h hp v tit niu. Dng phi hp khc l fansidar bao gm sulfadoxin v pyrimethamin c hiu qu trong iu tr st rt, t hiu qu i vi bnh nhim trng . 2.5.2. Cc beta- lactamin Cc thuc ny trong cng thc ha hc c cha vng beta-lactam bao gm cc penicillin v cc cephalosporin. 2.5.2.1 Cc penicillin: Cc penicillin c nhn chung l axit 6 amino-penicillanic hp bi mt vng thiazolidin v mt vng -lactam cc penicillin ny khc vi penicillin khc ch gc R gn vo nhn chung. Cc penicillin dng trong iu tr thuc 3 nhm chnh : - Cc penicillin c hot tnh cao i vi vi khun gram dng, b ph hy bi enzym penicillinase: penicillin G, penicillin V (phenoxy methylpenicillin), benzathin penicillin. - Cc penicillin c hot tnh i vi vi khun gram dng, khng i vi enzym penicillinase: methicillin, nafcillin, oxacillin, dicloxacillin . - Cc penicillin c hot tnh rng i vi c vi khun gram dung v vi khun gram m, b ph hy bi enzym penicillinase. Cc penicillin nhm ny bao gm: + Cc aminopenicillin: ampicillin, amoxicillin, becampicillin, metampicillin. + Cc carboxy penicillin: carbenicillin, ticarcillin. + Cc ureidopenicillin: azlocillin, mezlocillin, piperacillin.
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Ngoi 3 nhm chnh trn hin nay ngi ta cn c cc penicillin phi hp trong ngi ta kt hp cc penicillin khc nhau vi nhng cht c ch enzym lactamase dng iu tr cc nhim trng do vi khun khng sn xut enzym lactamase. Hai cht c ch enzym ny c dng phi hp vi cc penicillin l axit clavulanic v sulbactam: cc thuc ca s kt hp ny nh: timentin gm ticarcillin v axit lavulanic, augmentin (amoxicillin v a.clavulanic), unasyn (ampicillin & sulbactam). 2.5.2.2 Cc cephalosporin:
Cc cephalosporin c mt nhn chung l axit faminocephalo-sporanic hnh thnh

bi mt vng lactam kt hp vi mt vng dihydrotiazin thay th hydro cu nhm amin bng cc gc R khc nhau th thu c cc cephalosporin bn tng hp . Cc cephaloporin c hot tnh vi c vi khun gram dng v vi c vi khun gram m . Da vo tnh khng khun cc cephalosporin c chia cc th h: - Cc cephalosporin th h th nht: Bao gm cephalothin, cephazolin, cephalein, cefaclor. Cc thuc ny c hot tnh i vi t cu lin cu A, ph cu, E. coli, Klebsiella, Proteus.. - Cephalosporin th h th hai: Cc thuc nhm ny c hot tnh tt hn cc thuc cephalosporin th h th nht chng li E. coli, Klebsiella, Proteus, Citrobacter, Enterobacter, H.influenzae, N.gonorrhoe, N. meningitidis. Cc thuc nhm ny gm: cefamandol, cefuroxime, cefonicid, cefoxitin... - Cephalosporin th h th ba: C ph tc dng khng khun bao trm ton b vi khun ng tiu ha, tuy nhin c tc dng km vi vi khun gram dng, thuc nhm ny c th chia 02 nhm, nhm c tc dng chng Pseudononas aeruginosa gm ceftazidime v cefoperazone, v cc nhm khng c tc dng chng P. aeruginosa bao gm cefotaxim, ceftazidime, ceftizoxime, moxalactam. Nhiu thuc nhm ny thm vo mng no tt nn c dng iu tr vim mng no do vi khun gram m. 2.5.2.3. Monobactam v Carbapenem: Hai nhm thuc ny khc vi cc beta-lactam v cu to ha hc nhng c ch tc dng l c ch tng hp vch tng t nh cch tc dng ca cc beta lactam, aztreonam l thuc thuc nhm monobactam c sn hin nay, ph khng khun gii hn trn vi khun gram m tng t nh cc aminoglycoside. Imipenem l khng sinh carbapenem c sn hin nay, ph khng khun ca thuc ny c xem l rng nht trong cc thuc khng sinh hin nay bao trm ln cc vi khun gram dng gm c lin cu D, c tc dng chng li hu ht cc vi khun gram m k c P.aeruginosa, v c tc dng chng li vi khun k kh. 2.5.3. Cc khng sinh aminoglycoside Gm streptomycin, neomycin, gentamycin, tobramycin, amikacin, kanamycin
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Cc khng sinh nhm ny c tc dng dit khun bao gm c vi khun gram dng v nhiu vi khun gram m. gentamycin, tobramycin, neomycin c tc dng chng P.aeruginosa, tri li streptomycin c tc dng vi vi khun lao nn l thuc dnh ring iu tr lao. 2.5.4. Cc thuc tetracyclin C cng thc ha hc tng t nhau. Cc thuc khc nhau ch cc gc R khc nhau. Cc thuc ny bao gm tetracyclin, chlotetracyclin, oxytetracyclin, doxycyclin, minocyclin, thuc nhm ny c tc dng cn khun vi vi khun gram dng v gram m, cc tetracyclin c hiu qu vi Rickettsia, Chlamydia, Mycoplasma pneumoniae v Leptospira. 2.5.5. Chloramphenicol Lc u chit xut t Streptomyces venezuelae nhng hin nay c tng hp. Thuc ny c tc dng cn khun v ph khng khun rng. N c hiu qu trong bnh thng hn v cc nhim trng do Rickettsia. 2.5.6. Macrolit v cc thuc tng t Cc thuc khng sinh macrolit gm erythromycin, oleandomycin, spiramycin. Cc khng sinh thuc nhm ny c tc dng cn khun, chng tc dng tt trn vi khun gram dng, mt s vi khun gram m nh phy khun t. Erythromycin l thuc an ton nn c dng lm thuc thay th penicillin trong cc nhim trng do vi khun gram dng bnh nhn d ng vi penicillin. Lincomycin v clindamycin c c ch tc dng v hat ph tng t nh erythromycin. 2.5.7. Polypeptid Khng sinh nhm polypeptit ch yu l polymycin, ph tc dng ch yu dit khun vi vi khun gram m k c Pseudomonas. Polymycin c vi thn kinh v thn, hin nay t c dng ng ton thn iu tr cc nhim trng gram m. Cac thuc c polymycin thng c dng iu tr nhim trng ti ch. 2.5.8. Rifamycin L khng sinh chit sut t Streptomyces mediterranei, cht dn xut bn tng hp l rifampin, hai thuc ny c hat ph i vi mt s vi khun gram m v gram dng, vi khun ng rut, vi khun khng acid cn. Rifamycin do vy l thuc s dng ch yu iu tr bnh lao hin nay. 2.5.9. Cc quinolon v cacboxy - fluoroquinolon Hp cht quinolon tng hp u tin l axit nalidixic ch yu dng iu tr nhim trng ng tit niu. Cc cacboxy-fluoroquinolon l cc dn cht t axit nalidixic, cc thuc ny c tc dng dit khun i nhiu vi khun gram dng v vi khun gram m k c t cu khng vi methicillin v P.aeruginosa, chng cn cn c tc dng chng Mycoplasma v Chlamydia. Cc fluoroquinolon c s dng trong nhiu nhim trng cc c quan. Mt s fluoroquinolon thng dng hin nay gm ciprofloxacin, enoxacin, norfloxacin, pefloxacin.
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2.6. S khng ca vi khun i vi khng sinh Theo di s khng ca vi khun i vi khng sinh bng khng sinh ngi ta nhn thy khi mi s dng cht khng sinh, ngi ta t gp cc chng khng thuc. Nhng dn d khng sinh c s dng lu di v ph bin. Ngi ta nhn thy c nhiu chng vi khun chng li c khng sinh, thm ch c loi vi khun ph thuc vo khng sinh sinh trng v pht trin. V d: T cu vng, vi khun l i vi khng sinh Streptomycin. Hin nay ngi ta pht hin c hai c ch khng cht khng sinh: 2.6.1. S khng cht khng sinh c ngun gc t nhim sc th: Ngi ta phn lp c cc chng khng thuc t qun th nhy cm bng cch phun huyn dch vi khun ln trn b mt a thch c ha tan mt nng khng sinh sc c ch vi khun, nhng sau 24 gi nui cy c th tm thy mt vi khun lc mc ln c. Ngi ta cho rng, nhng khun lc xut hin ny y l mt bin c him, xut hin vi tn s 10-9. Bin c ny c tnh cht bn vng v truyn li c cho th h sau. S t bin ca vi khun khng vi khng sinh ny khng lm thay i xc sut ca vi khun i vi s t bin vi khun khng vi khng sinh khc. Do , xc sut vi khun mt lc khng hai loi khng sinh bng tch s xc sut ca mi t bin. V d: T cu khng penicillin vi xc sut 10-9, khng Streptomycin-10-9. Mun c t cu khng c penicillin v streptomycin tn s xut hin l 10-18. Do , ta c th ni rng hin tng khng cht khng sinh khng phi do cht khng sinh cm ng v gy nn, khng phi l hin tng thch ng ca vi khun i vi mi trng. Trn thc t, khng sinh ch tc ng nh mt nhn t chn lc git cht i ng o vi khun nhy cm, ch li nhng chng khng khng sinh sinh trng v pht trin thun li hn. 2.6.2. S khng cht khng sinh c ngun gc ngoi nhim sc th : Bn cnh ADN trong nhim sc th gi vai tr ch yu trong s di truyn. Tnh khng cht khng sinh vi khun cn c yu t ADN ngoi NST gi l plasmit, nhng plasmit ny c th ci xen vo NST hay nh km NST ca vi khun ch v c sao chp cng vi NST. Khi dng rng ri khng sinh trong iu tr s lm tng s chn lc cc vi khun khng li khng sinh hay s ly chuyn cho ca cc vi khun khng cht khng sinh thng qua mt trong 3 c ch: bin np, ti np v tip hp. Cc phng tin lm ly lan nhanh tnh khng thuc - Plasmid: thm vo cc gen khng thuc, trn cc plasmid cn mang cc gen thc y s trao i gen gia cc vi khun. - Do transposon: y l nhng gen c kh nng di chuyn. Cc transposon c mang nhng gen khng khng sinh c th nhy t plasmid ny n mt plasmid khc hoc t
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plasmid qua nhim sc th. Mt s transposon khng thuc tm thy vi khun Gram m cn mang gen thc y s ly truyn. - Do cc integron y l vector c th mang gen khng khng sinh, integron c th chn vo cc v tr nht nh trn nhim sc th 2.7. Qui tc s dng khng sinh: Ch dng khng sinh khi cn thit, nn chn khng sinh thch hp cn c theo khng sinh . Phi s dng phi hp cc loi khng sinh trong khi cha bnh, c bit i vi vi khun c t l bin chng cao. Dng khng sinh phi liu lng, thi gian cn thit.
3. Vitamin
3.1. S lc v vitamin Vitamin l mt nhm cht hu c c phn t tng i nh v c bn cht l ha hc rt khc nhau. Nhm cht hu c ny c bit cn thit cho hot ng sng bnh thng ca cc c th sinh vt d dng. Trong nhng nm gn y ngi ta chng minh rng vitamin cn cn thit cho cc c th t dng nh thc vt. Ngi u tin bit s dng thc phm giu vitamine cho thu th l James Cook cho thy th n rau u v choucroute (mt loi da chua lm bng bp ci). Nh vy m h khng mc chng scorbut trong cuc hnh trnh nhiu thng. T th k th XVIII , James Lind, chng minh rng dng nc ct tri chanh hay cam s nga cn bnh tn ph cc thy th m ngy nay hu nh ai cng bit l do thiu vitamin C. Jean Baptiste Dumas, nh ha hc kim chnh tr, ch ra mt loi sa nhn to. ng pha trn cht bo, albumin trong nc ng. Tuy khng ngon nhng v mt dinh dng sa nhn to ny c glucid, lipid v protein. Nhng ch y v mt nng lng m vn cn thiu mt cht cn thit no . Louis Pasteur l ngi u tin bit s dng cc mi trng tng hp nhn to nui cy VSV Khong 1859-1864, Pasteur thm mt cht nc chit ca cc cht hu c t nhin vo th thy nm men pht trin mnh hn ln. Nm 1888 N.I.Lunin nui chut bng sa nhn to c dinh dng cao (lm t cazein tinh khit, lactose, lipid, mui khong) sau mt thi gian ngn chut b cht ngoi cc cht trn trong sa cn c cc cht khc khng th thiu c v mt dinh dng Frederick Gowland Hopkins(1861-1947), sau 6 nm nghin cu (1906-1912) ng d rt ra kt lun: Nu khng b sung cc cht c ngun gc t nhin (sa, nc chit c ci, nc chit nm men...) th khng con chut no sng qa 4 tun.
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Nm 1897 ikman chng minh c nguyn nhn gy bnh n ph, vim dy thn kinh g l do n go xay bng my. Nu dng cm b sung vo khu phn nui chut khc phc c bnh ny. Nm 1911 nh khoa hc M gc Balan tch ra c t cm mt cht kt tinh khng mu. B sung vo t.n nui b cucha bnh vim dy thn kinh, ng t tn cht amin sng hay Vitamin. Nm 1913 Collum v cng s tm thy mt loi VTM tan trong cht bo (du) v t tn l nhn t A, Nm 1916 bt u c gi l vitamin A - Tc dng ca Vitamin A Vitamin A c nhiu trong gan ng vt,lng trng, trong cc loi rau, c, qu mu (gc, ng, , c chua, c rt.. Gi cho da khng b kh, khng ha sng khng vi s xm nhim ca VSV - Vitamin C: Nh khoa hc M gc Hungari gii Nobel 1937 l ngi u tin xc nh cng thc ha hc ca VTM C v chc nng ca n. - Vitamin D c nhiu trong gan ng vt, du c - Vitamin K, c pht hin nm 1929 , C vai tr trong ng mu. Thiu K mu khng ng. 1.2. Cc nhm vitamin -Vitamin tan trong acid bo 1. 2. 3. 4. 5. 6. Vitamin A v caroten Vitamin D (canxipherol) Vitamin E (tocopherol) Vitamin K (philoquinol) Vitamion Q (ubiquinol) Vitamin F (phc hp cc acid bo khng no)
-Vitamin tan trong nc 1. Vitamin B1 (tiamin) 2. Vitamin B2 (riboflavin) 3. Vitamin B6 (piridosin) 4. Vitamin PP (acid nicotinic) 5. Vitamin C (acid ascorbic) 6. Vitamin B12 (xiancobalamin), H(biotin), P(rutin) - Cc vitamin t c nghin cu nh vitamin Bt(carnitin), coli,inosit, B13(acid orotic)
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Cu hi n tp chng 12
1.Th no l bnh truyn nhim ? Ngun gc, c im ca bnh truyn nhim ?2.Vi sinh vt gy bnh c th lan truyn theo nhng con ng no ?3.Cho bit tc nhn gy nn cc bnh thng gp nh: bnh lao, bnh ng c tht, Bnh bi lit, bnh than ? Cho bit cch phng v chng cc bnh trn.4.Phn bit min dch c hiu v khng c hiu, min dch dch th v min dch t bo, Bnh l min dch.5. Hy phn bit min dch c hiu v khng c hiu (kh nng khng). Min dch c hiu gm nhng loi min dch no ? 6. Hy phn bit cht sinh min dch, khng nguyn v hapten. Hy nu tnh cht c bn ca khng nguyn. Quyt nh khng nguyn (epitop) l g ? 7. V s cu to khng th. Vng no ca khng th gi vai tr gn khng nguyn. Cc lp khng th c chc nng g ? 8. K tn cc t bo tham gia vo p ng min dch. Nu s khc nhau c bn gia t bo T v B. 9. Phn t MHC l g ? chng c chc nng g trong trnh din khng nguyn ? 10. Hy trnh by qu trnh trnh din khng nguyn vi s tham gia ca MHC-I v MHC-II. 11. Hy trnh by qu trnh hnh thnh khng th. Ti sao c th c tim nng to ln trong vic hnh thnh khng th chng li mi khng nguyn c th c trong t nhin. 12. B th c hot ho nh th no v ng vai tr g trong p ng min dch.
* Ti liu c thm
* Ti liu tham kho

* Gii thch thut ng

Bch cu, hay bch huyt cu (ngha l "t bo mu trng", cn c gi l t bo min dch), l mt thnh phn ca mu. Chng gip cho c th chng li cc bnh truyn nhim v cc vt th l trong mu. Chng l mt phn ca h min dch, s lng bch cu cc loi trong mt lt mu ngi ln khe mnh dao ng t 4x109 ti 11x109.
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Cc bnh min dch gm ba nhm ln: suy gim min dch, trong mt vi b phn ca h min dch khng to ra c p ng mnh (th d bnh bch cu ht mn tnh); cc bnh t min, khi h min dch tn cng cc khng nguyn ca chnh c th mnh (lupus ban h thng, vim a khp dng thp, bnh Hashimoto bnh nhc c v.v.); v cc phn ng qu mn, trong h min dch p ng khng thch ng vi cc vi cc hp cht v hi (hen ph qun v cc d ng) hoc p ng qu mnh. T bo lympho L cc t bo chuyn bit ca h min dch, cc t bo lympho (lymphocyte) rt ph bin trong h bch huyt. Trong mu c ba loi lymphocyte: t bo B, t bo T v cc t bo git t nhin (natural killer (NK) cell). Cc t bo B sn xut ra khng th lin kt vi tc nhn gy bnh nhm to iu kin c th ph hy chng. Cc t bo T CD4+ (T b tr) phi hp cc phn ng ca h min dch (loi t bo ny b suy gim khi c th b nhim virus HIV). Cc t bo T CD8+ (T gy c) v t bo git t nhin c kh nng git cc t bo ca c th b nhim cc tc nhn gy bnh ni bo. T bo mast: Bit ha t cc bch cu i toan trong mu v ti cc m, t bo mast l loi t bo tun tra khp c th v c trang b nhng v kh li hi: cc ht trong bo tng ca chng cha nhng cht c tng hp sn gm histamine, serotonine, v nhiu enzym. T bo mast sn xut cc eicosanoid gy gin mch v cc PAF (platelet activating factor) kch hot tiu cu. Cc cht trn c tng hp kh nhanh chng t cht bo. T bo mast cn sn xut cc cytokine vim: IL-1, TNF v cc yu t ha hng ng (chemokine). Trn b mt t bo mast c cc th th ca IgE, IgG v cc mu b th, khi pht s phng thch cc thnh phn cha trong cc ht ca chng khi c kch hot. T khng th u th k 20, Paul Ehrlich 1854-1915 a ra khi nim horror autotoxicus (tm dch "tnh t c ng s"), cho rng h min dch khng to ra khng th chng li cc thnh phn ca chnh c th, v iu ny s dn n t hy dit. Quan nim ny c chp nhn rng ri sut gn trn th k cho n khi khi nim "mng li khng th" ra i. u thp nin 1980, ngi ta khm ph ra cc t khng th hnh thnh t pht vi s lng t, thng c hiu vi nhiu khng nguyn ca c th nn gi l a c hiu. Cc t khng th ny kh lnh, khng gy phn ng hy dit nh cc t khng th trong cc bnh t min, khi c ch iu ha min dch b qua mt. H vi sinh vt t nhin da - m thp v pH acid
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H VSV ch yu l cc vi khun Gram dng, hin din ni c m cao nh tuyn m hi, tuyn nhn: Staphylococcus, Coryne-bacterium, Acetinobacter, Pityrosporum, Propionibacterium, Micrococcus H vi sinh vt vng ming - Streptococcus, Lactobacillus, Fusobacterium, Veillonella, Corynebacterium, Neisseria, Actinomyces - Su rng do hot ng ca VSV: + Streptococcus sobrinus v S. mutans hnh thnh lp polysaccharide (glycocalyx) gip gn vo b mt rng to thnh vt tan rng (dental plaque), tng trng lm cn kit oxygen + Vi khun lactic acid to ra acid hu c lm gim pH b mt rng, lm tan thnh phn calcium phosphate ca rng, gy ra su rng. H vi sinh vt ng tiu ha - Lactobacillus, Streptococcus, Bacteroides, Bifidobacterium, Eubacterium, Peptococcus, Peptostreptococcus, Ruminococcus, Clostridium, Escherichia, Klebsiella, Proteus, Enterococcus, Staphylococcus - VSV k kh v k kh ty - VSV c li - VSV ln men thi - VSV c li: cung cp vitamin, h tr bin dng vt ch.
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CHNG 13
SINH THI HC VI SINH VT I. I CNG V SINH THI HC VI SINH VT

1. Cc khi nim c bn
Sinh thi hc (ecology): nghin cu s hnh thnh tn ti v pht trin ca mt h thng cc sinh vt cng vi cc iu kin khng sng nht nh trong mi quan h hu c tc ng qua li vi nhau. Sinh thi hc vi sinh vt: nghin cu vi sinh vt v kha cnh sinh thi hc - Mc tiu ca sinh thi hc vi sinh vt: + Nghin cu s a dng sinh hc (chng loi, s lng) ca vi sinh vt trong t nhin, s tng tc ca cc qun dng khc nhau trong qun x + Nghin cu hot tnh ca vi sinh vt trong t nhin v gim st tc ng ca vi sinh vt ln h sinh thi. ngha ca nghin cu sinh thi hc VSV - Vi sinh vt c vai tr thit yu cho s duy tr pht trin ca cc h sinh thi: + Thu ly nng lng nh sng, c nh m, c nh CO2, to O2,phn hy cht hu c + L tc nhn chnh thc hin cc phn ng trong chu trnh sinh a ha cc nguyn t cn cho s sng + L tc nhn gip phn hy c cht, phc hi mi trng - Sinh thi hc VSV gip hiu c tng tc ca VSV vi nhau v vi mi trng, hiu vai tr ca VSV trong iu kin t nhin ca cc h sinh thi
2. c im ca vi sinh vt trong t nhin

Mi trng sng ca vi sinh vt (Habitat, niche, microenvironment): + Habitat: mi trng sng ni cc qun th, qun dng hnh thnh qun x + Niche: l vi mi trng ti u cho s tng trng ca vi sinh vt - Trong t nhin, vi mi trng (microenvironment) l ni m vi sinh vt thc t sng v bin dng: + Cc iu kin ha l ca vi mi trng bin i rt nhanh theo khng gian v thi gian + Trong mt khng gian vt l hp c s tn ti nhiu vi mi trng khc nhau + Tnh khng ng nht ca vi mi trng quyt nh tnh a dng ca vi sinh vt Mng sinh hc (biofilm)
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- Cc b mt mng c dng lm ni sng ca VSV, c cc cht dinh dng bm vo, cc VSV bm vo v s chuyn ha cc cht tt hn nhiu so vi trong nc t do do hiu qu ca s hp ph. - Cht dinh dng l nhn t hn ch tc tng trng trong hu ht cc mi trng t nhin v c cung cp dng ngu nhin. - Vi sinh vt thng hin din trn b mt mt gi th do nng cht dinh dng gii hn y cao hn to thnh mng sinh hc (biofilm) hoc tp hp cc khun lc ca cc qun th khc nhau. Tng trng ca VSV trong t nhin - Cht dinh dng trong t nhin - ti nguyn (resources) cho VSV khng c cung cp lin tc - VSV tng trng thnh xung theo ngun ti nguyn ca mi trng - Cc cht d tr trong t bo (PHA, PHB, polysaccharide, polyphosphate) c s dng khi ngun ti nguyn b cn kit - c im chung ca tng trng ca VSV trong t nhin: + Tng trng hm m thng ngn + Tc tng trng nh hn rt nhiu so vi trng hp nui cy thun chng trong phng th nghim - Tc tng trng chm do: + Ngun cht dinh dng thp + Phn b cht dinh dng khng ng u + B cnh tranh bi cc qun th khc S hnh thnh qun x vi sinh vt Cc mc t chc ca vi sinh vt trong t nhin + T bo + Qun th (population): tp hp cc t bo cng loi, c hnh thnh do s tng trng ca cc t bo ring bit trong mt vi mi trng nht nh + Qun dng (guild): tp hp cc qun th khc loi c c tnh chung v ngun cht dinh dng, cc yu t ho l trong mt vi mi trng + Qun x, h vi sinh vt (community): tp hp nhiu qun dng cng hin din trong mt iu kin mi trng, tin hnh nhng qu trnh sinh l b tr nhau cng tng trng + H sinh thi (ecosystem): nhiu qun x c hnh thnh c mi quan h vi nhau v nng lng v vt cht
3. Cc phng php nghin cu sinh thi hc vi sinh vt

Phng php nghin cu tnh a dng
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Phng php nui cy - Nui tch ly (enrichment culture): dng mi trng chn lc v iu kin tt u lm tng t l hin din tng i ca vi sinh vt mc tiu t mt ngun t nhin - Phn lp v lm thun vi sinh vt mc tiu Nui tch ly - Nui tch ly = nui lm giu - Nui tch ly trong h kn: + Mi trng chn lc lng, iu kin ha l chn lc, ngun phn lp thch hp + Ch thu c cc qun th tng trng nhanh ca mt qun dng - Nhc im ca phng php nui tch ly trong phn tch tnh a dng: qun th thu c trong nui tch ly khng chc l qun th chim u th trong mu Nui tch ly bng ct Winogradski - Ct Winogradski: l mt h sinh thi k kh nh xu c dng nh mt ngun sng lu di u vit i vi tt c cc sinh vt nhn s tham gia vo vng tun hon cc cht dinh dng + Bn, nc ao h c b sung cht hu c quan tm + Thu c nhiu qun th a dng ca nhiu qun dng. Nui tch ly bng h n ha - H n ha (chemostat): + Nui cy lin tc trong mi trng lng cha ngun carbon quan tm + Lm giu v phn lp qun th bin dng c cht - Phn lp v lm thun + Hp ria (streak plate): phn lp cc chng hiu kh hoc k kh khng nghim ngt + Pha long ti hn (extincting dilution): phn lp cc chng hiu kh + Pha long trong ng thch mm (agar shake tube method): phn lp cc chng k kh - Cc phng php pha long c th c dng xc nh qun th chim u th trong mu nh danh chng thun - Phng php sinh ha, min dch - Phng php gii trnh t SSU-rRNA (small subunit ribosomal RNA) - Phng php lai phn t vi mu d chuyn bit - Phng php vi mu d pht sinh loi (phylogenic probe) - Phng php FISH (fluorescent in situ hybridization) - K thut FISH: lai phn t trn mu t nhin bng mu d c nh du hunh quang - Mu d:
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+ Mu d pht sinh loi: cc trnh t nhn din vi sinh vt cc mc phn loi khc nhau + Mu d chuyn bit i vi mt gen mc tiu - S dng ng thi nhiu mu d pht sinh loi c nh du hynh quang khc nhau. ng dng ca FISH trong nghin cu sinh thi hc vi sinh vt - Phn lp vi sinh vt (k thut Nhp quang hc) - nh tnh v nh lng vi sinh vt - Nghin thnh phn v tng tc ca vi sinh vt: + S dng ng thi nhiu mu d pht sinh loi c nh du hynh quang khc nhau: quan st v c trng ha thnh phn ca vi sinh vt + Kt hp FISH vi confocal laser microscope: quan st v c trng ha thnh phn, s tng tc ca cc nhm vi sinh vt khc nhau trong mu t nhin + i vi cc vi sinh vt cha th nui cy: m t hnh thi, mt v tng tc vi cc sinh vt khc - Nghin cu hot tnh ca vi sinh vt: k thut ISRT Phn lp bng k thut nhp quang hc - Nhp quang hc (Optical tweezer): + Knh hin vi o c trang b mt ngun laser hng ngoi mnh v b vi thao tc cha ng mao qun + T bo mc tiu c nh du hunh quang s c pht hin, tch khi cc t bo khc trong ng mao qun bng lc ca chm nh sng laser + Ct ng mao qun v thu nhn t bo mc tiu - ng dng: + Phn lp cc chng vi sinh vt c tc tng trng chm + Phn lp cc vi sinh vt thiu s trong qun x + ng dng khc: phn lp, tch nhm cc phn t mc tiu, nh v trng v tinh trng trong th tinh in vitro... Pht hin vi sinh vt bng k thut Nhum nhim sc th - Nhum NST (Chromosome painting): + Gen, mt nhm cc gen, mt b gen c ct thnh nhng an mu d 50 200bp v nh du hunh quang + Lai in situ v dng k thut knh hin vi hunh quang pht hin vi sinh vt mc tiu - ng dng: + Tm s hin din ca mt nhm sinh l (qun dng) trong mi trng (vi sinh vt c mang mt gen, mt nhm gen nht nh: nitrogenase, cc thnh phn ca h quang...)
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+ nh lng nhm sinh l Xc nh thnh phn ca qun x bng gii trnh t SSU-rRNA - K thut gii trnh t rRNA c dng trong sinh thi hc vi sinh vt phn tch thnh phn cc qun x vi sinh vt m khng cn phn lp, nui cy chng vi sinh vt - Qui trnh: + Thu nhn DNA tng + Dng cc cp mi chuyn bit (Bacteria, Archea, Eukarya, hoc ca cc mc phn loi thp hn) khuch i SSU-rRNA + Lp ngn hng cc dng SSU-rRNA + Tuyn chn cc dng mc tiu bng lai in situ vi mu d pht sinh loi + Gii trnh t SSU-rRNA + Lp cy pht sinh loi nh lng bng phng php truyn thng - nh lng mt nhm vi sinh vt chn lc - Nui cy trn mi trng chn lc v m khun lc - Phng php MPN: + Pha long ti hn mu t nhin (pha long bc 10) + Lp cc dy ng th nghim c lp li 3 hoc 5 ng vi 3 hoc 5 pha long bc 10 + nh tnh s hin din ca nhm vi sinh vt mc tiu da trn cc tn hiu d quan st + Tra bng MPN nh lng bng phng php nhum hunh quang - Dng phm nhum hynh quang v knh hin vi hunh quang - Tn hiu nn thp cc mu t nhin - nh lng tng t bo: + Nhum hynh quang v m di knh hin vi hynh quang: acridine orange, DAPI (4',6-diamido-2-phenylindole) nhum nucleic acid + Khng phn bit t bo sng - cht nh lng bng khng th hunh quang - nh lng bng khng th pht hunh quang: + Dng khng th c nh du hunh quang + Pht hin vi sinh vt mc tiu, tnh chuyn bit cao - nh lng bng k thut FISH nh lng bng mu d pht sinh loi hunh quang Phng php nghin cu hot tnh vi sinh vt trong t nhin
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Cc phng php truyn thng - Thng thng l cc phng php ex situ: mu c thu, bo qun v xc nh hot tnh ti phng th nghim - o hot tnh tng ca vi sinh vt da trn mc tng trng: + Tng O2 tiu th, tng CO2 phng thch + Tng ATP - o tng hot tnh cc enzym ch yu trong cc chuyn ha vt cht do mt hoc mt s qun dng Cc phng php in situ - Phng php in situ: o t hot tnh ca vi sinh vt ngay trong mi trng v vi mi trng - Phng php o in situ tng hot tnh da trn h hp, ATP, o hot tnh chuyn ha - Phng php ng v phng xa: o hot tnh chuyn ha vi nhy cao + Hot tnh ring (specific activity): t l hot tnh trn c cht ng v so vi hot tnh tng + Loi b hot tnh do phn ng phi sinh vt: i chng t bo cht (killed cell control) - Phng php vi in cc (microelectrode): theo di bin i v thnh phn ha hc trong vi mi trng + Cc vi in cc pH, oxygen, N2O, CO2, H2, H2S + Di chuyn theo cc bc 1mm bng micromanipulator + Dng nghin cu cc chuyn ha trong cc li sinh khi (micromat) ng dng FISH nghin cu hot tnh ca qun dng trong k thut ISRT - ISRT (In situ Reverse transcriptase): + Lai in situ mRNA bng primer chuyn bit + Thc hin phin m ngc to cDNA v khuch i bng PCR + Lai vi mu d nh du hunh quang - ng dng: + Nghin cu xc nh vi sinh vt c biu hin gen trong qun x mt thi im nht nh + Xc nh qun th c hot tnh trong qun dng v qun x + Nghin cu nh hng ca cc yu t mi trng ln s biu hin ca gen trong qun x X nh hot tnh vi sinh vt bng ng v bn - ng v bn (stable isotope): ng v bn mang nhiu neutron hn dng bnh thng
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- Hiu ng phn an ng v (isotope fractionation): phn ng sinh ha c xu hng dng ng v nh hn ng v nng, lm gim t l tng i ca ng v nng so vi ng v nh trong sn phm - T s ng v - ng dng: + Xc nh thi im xut hin s sng da vo t s ng v C trong c + Xc nh thi im mi trng tri t t trng thi kh sang trng thi oxygen ha (o/oo): t s ng v nng so vi ng v nh trong vt cht - Dng ng v bn dng trong nghin cu sinh thi hc vi sinh vt: 13C, 34S, 18O
4. Hot ng v vai tr ca vi sinh vt trong cc h sinh thi

4.1. Vi sinh vt trong cc mi trng trn cn - Vi sinh vt tng trng ch yu trn b mt cc ht t - a dng cao v tn ti nhiu vi mi trng khc nhau - Cc phng php nghin cu: + Tm s hin din ca vi sinh vt: nhum bng cam acridine, DAPI, quan st bng knh hin vi hynh quang + Tm mt vi sinh vt mc tiu: dng k thut min dch hunh quang hoc FISH + Quan st trng thi hin din trn b mt gi th: knh hin vi in t qut SEM + Cc phng php xc nh hot tnh ca vi sinh vt 4.2. Yu t nh hng n hot tnh vi sinh vt trong mi trng t - m: nh hng mnh n hot tnh vi sinh vt + Thay i vi phm vi rng trong mi trng nc + nh hng n oxygen trong t: t ro nc c nng oxygen khuch tn cao, t nhiu nc thng lm mi trng k kh - Cht dinh dng: nh hng mnh n mt v hot tnh vi sinh vt + Cht dinh dng hu c tp trung b mt cc ht t v vng quanh r cy + Cht dinh dng gii hn thng l cc cht dinh dng v c, c bit l P, N 4.3. Vi sinh vt cc tng su - Mu t su n 300m c s a dng cao ca cc vi sinh vt k kh, k kh ty v hiu kh: + Cht dinh dng t nc ngm thm qua + Hot tnh bin dng thp hn rt nhiu so vi vi sinh vt trn mt t - Mu t su 1.500m c s a dng ca vi khun k kh: + Chng minh c hot ng ca cc dng ha nng v c (kh sulfate, sinh methane, sinh Acetate ng hnh) bng phn tch t s ng v trong CH4 + Cht cho in t l H2, cht nhn in t l CO2, SO42-
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+ Ngun to cht cho in t phn ng ha hc gia H2O v FeO - ngha: + V c ha hp cht hu c to thnh phn ha hc ca nc ngm + In situ bioremediation cc cht nhim 4.4. Vi sinh vt trong h sinh thi ao h - Thnh phn v hot tnh vi sinh vt thay i theo tnh cht ha l ca mi trng nc: - Hu ht cc vi sinh vt tham gia vo cc chu trnh sinh a ha cc nguyn t u hin din - Vi sinh vt l thnh phn quang nng quan trng nht: + Quang hp sinh oxygen: to lam, to + Quang hp khng sinh oxygen: vi khun quang dng - Hnh thnh mt chui thc phm trong nng sut sinh hc ph thuc vo tc ca sn xut s cp - Cc yu t quan trng nh hng n thnh phn v hot tnh vi sinh vt: + O2 tan lm thay i mi trng v vn tc chuyn ha cht hu c + Cc cht v c (NO3-, PO43-) to s pht trin bng n ca to, nh hng n O2 tan 4.5. nh hng ca oxygen tan v cht hu c ln vi sinh vt trong mi trng nc - Oxygen v chu trnh carbon c quan h cht ch vi nhau: nng oxygen tan t l nghch vi nng cht hu c tan - BOD (biochemical oxygen demand) biu th lng cht hu c tan - Nng oxygen tan trong nc cn chu nh hng ca cc yu t thy triu, tc dng chy, nhit theo ma - Oxygen tan lm mi trng nc tr nn hiu kh hay ym kh, nh hng n thnh phn v hot tnh ca vi sinh vt, thnh vt thc, ng vt thy sinh v cht lng mi trng nc 4.6. Vi sinh vt trong bin su - c im ha l ca mi trng bin su: + su trn 1000m, chim hn 75% tng dung tch nc ca i dng + t cht dinh dng, khng c nh sng, lnh (2 - 30C), p sut thy tnh cao (+1atm/10m su) - c im ca vi sinh vt: + a hn (psychrophilic) v a hn cc oan (extreme psychrophilic) + Chu p (barotolerant): tng trng c p sut bnh thng cho n 300atm + a p (barophilic): tng trng ti u 400atm
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+ a p cc oan (extreme barophilic): ch tng trng c p lc tn 300atm, ti u 700 - 800atm - ngha ca nghin cu vi sinh vt trong bin su - Barophile l m hnh nghin cu nh hng ca p sut ln s biu hin ca gen, hot ng ca protein v hot ng ca mng - p sut cao: + Lm gim hot tnh ca enzym do lm gim i lc vi c cht + Lm tng thnh phn axt bo khng bo ha + Cm ng tng hp porin Omp H trn mng ngoi - Gen m ha Omp H t mt barophlie c cm ng biu hin mc phin m trong E. coli p sut 200atm: + Omp H l mt porin khc vi cc porin c tng hp 1atm + C ch cm ng s biu hin ca gen: bt hot repressor hoc hot ha actitvator bi p sut cao? 4.7. Vi sinh vt ti cc khe thy nhit - Khe thy nhit (hydrothermal vent): dng phun nc nng mang nhiu cht khong c to ra ti cc vt nt trong lng bin su: + Khe m: dng phun tc 0,2 - 5cm/giy c nhit 6 - 230C (so vi 20C) + Khe nng: dng phun tc 1 - 2m/giy mu en do cha nhiu khong cht, nhit 270 - 3800C - Vi sinh vt ti khe thy nhit: sinh vt sn xut bng ha nng v c + Mt cao ca vi khun ha nng v c oxygen ha H2, cc hp cht kh ca lu hunh, nitrogen, carbon, st + C kh nng c nh CO2 - S cng sinh ca vi khun Thiovulum trong cc dng th (trophosome) ca trn khng l Pogonophora: trn thu thp, cung cp O2, cc khong cht cho vi khun, vi khun cung cp cc dng cht cho trn - Chui thc phm ti cc khe thy nhit: - Chui thc phm hai thnh phn: + Sinh vt sn xut ha nng v c + Sinh vt tiu th: ng vt cng sinh vi sinh vt sn xut
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Hnh 13.1.Khe thy nhit
5. Vai tr ca vi sinh vt trong cc chu trnh sinh i ha - cc nguyn t cn cho s sng

5.1. Chu trnh carbon - Chu trnh C c quan h cht ch vi chu trnh O thng qua hat tnh b tr ca cc sinh vt t dng (c nh CO2 to O2) v sinh vt d dng (phng thch CO2, tiu th O2) - Cc d tr C trong t nhin: kh quyn, t, i dng, trm tch, v sinh khi - Tc lu chuyn C qua cc d tr rt khc nhau, tc cao nht l gia kh quyn v sinh khi do cc phng thc c nh CO2 ca t dng v h hp hu c hiu kh ca d dng
Hnh 13.2. Chu trnh carbon - S chuyn ha C qua th kh khc nhau CH4, (CH2O)n, CO2 trong iu kin c v khng c oxygen u c s tham gia ca vi sinh vt
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Hnh 13.3. Cc chuyn ha C trong iu kin c v khng c oxygen Cc chuyn ha c ngha sinh thi trong chu trnh C - C nh CO2 vi sinh vt quang nng v vi sinh vt ha nng v c l bc chuyn ha quan trng nht: + Hnh thnh cht hu c mi + Lun chuyn C t khu vc phi sinh hc (kh quyn) sang khu vc sinh hc (sinh khi) + Sinh quyn c hnh thnh trn c s hai chuyn ha C ngc nhau l quang hp v h hp, trong cn cn quang hp nng hn h hp - S phn hy sinh khi, cht hu c bi vi sinh vt thnh CO2: + Phn hy k kh cht hu c thnh CH4 v CO2 + Oxygen ha CH4 thnh CO2 + CO2 do s phn hy bi vi sinh vt chim phn quan trng nht trong s phng thch CO2 vo kh quyn Phn hy k kh cht hu c v vai tr ca s cng dng
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Hnh 13.4. Phn hy hp cht hu c trong iu kin k kh - Phn hy hp cht hu c trong iu kin k kh: + Thy phn i phn t thnh n phn + Ln men s cp to H2, CO2, cn v axt bo + Tiu th H2, CO2 v chuyn ha cng dng cn, axt bo + To CH4 - Cnh tranh tiu th H2 trong iu kin k kh: + To CH4 (methanogenesis), to Acetate (acetogenesis) v to H2S (sulfidogenesis) + Trong mi trng khng c sulfate: cnh tranh gia sinh methane v sinh Acetate (Methanogen chim u th trong d dy ng vt nhai li; Homoacetogen chim u th trong rut mi) + Trong mi trng c sulfate: vi khun kh sulfate chim u th - S cng dng (syntrophism): + C ngha quan trng trong chuyn ha k kh tip tc cc cht hu c l sn phm ca ln men s cp + Cng dng gia syntroph v cc loi methanogen tiu th H2
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5.2. Chu trnh nitrogen - Nitrogen l mt nguyn t thit yu trong sinh cht - Trong t nhin hin din cc th kh khc nhau: NH3, NH2-, N, N2O, NO, NO2-, NO3-. D tr nitrogen quan trng nht l N2 trong kh quyn - S chuyn ha qua li gia cc dng ny cn c vai tr ca vi sinh vt
Hnh 13.5. Chu trnh Nitrogen
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- C nh nitrogen: + c thc hin bi mt s vi sinh vt c nh m t do v cng sinh + Chuyn N t dng khng th s dng bi a s sinh vt thnh dng c th s dng c + Dng c nh NH3 c chuyn t khu vc phi sinh vt vo sinh khi (NH2-) bi vi sinh vt, thc vt - Ammn ha: NH3 c to ra do s phn hy N hu c + NH3 c ti s dng bi thc vt, vi sinh vt, tn ti bn dng NH4+ trong iu kin k kh hp ph mnh bi cc ht t, t tan trong nc - Nitrate ha NH3: NH3 b oxygen ha bi vi khun nitrite ha v nitrate ha thnh NO2 v NO3-
+ NO3- tan tt trong nc v d b ra tri, lm tht that m trong t - Phn nitrate ha: + NO3- b kh bi vi sinh vt k kh thnh N2 lm tht that m trong t + Gim NO3- trong nc, cn tr hin tng n hoa ca to lm nhim nc 5.3. Chu trnh lu hynh - Lu hunh tn ti ch yu ba dng th kh S2-, S0 v S+6 - D tr ch yu dng CaSO4 v FeS2 - Cc phn ng chuyn ha cc dng ca S l do vi sinh vt v cc phn ng ha cht - H2S: + c to ra bi s kh sulfate v bi s phn hy cc amino acid cha -SH: + Mi trng k kh c s hin din ca sulfate v cc cht hu c + C c tnh i vi sinh vt hiu kh - Oxygen ha cc hp cht sulfide: + H2S b oxygen ha ha hc trong iu kin c oxygen + Trong mi trng vi hiu kh b oxygen ha bi vi sinh vt + Trong mi trng c nh sng, khng c oxygen, b oxygen ha k kh bi vi khun quang hp - Oxygen ha lu hunh bi vi sinh vt thnh SO42- v H+ lm gim pH - Chuyn ha ca lu hunh hu c (dimethyl sulfide) bi vi sinh vt: + Cht cho in t to lc kh vi khun ta + Cht cho in t thu nng lng + Cht nhn in t trong h hp k kh + C cht ln men sinh methane
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Hnh 13.6. Chu trnh lu hunh
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5.4. Chu trnh st - Hai dng th kh chnh ca st trong t nhin l Fe2+ v Fe3+ ph thuc vo pH v O2 - Fe3+: ch tan trong nc pH axt hoc dng phc hp vi cc hp cht hu c; b kh thnh Fe2+ bng phn ng ha hc hoc bi vi sinh vt - Fe2+ b oxygen ha bi O2 thnh Fe3+ + Bn trong iu kin khng c O2 hoc trong mi trng c O2 pH axt + Trong khng kh pH axt, Fe2+ l cht cho in t ca vi sinh vt (Thiobacillus ferrooxidans) to Fe3+ Chu trnh st v hin tng nc r cha axt cc m khong + Pyrite FeS2 hin din trong qung khong b oxygen ha thnh Fe2+ v H2SO4 + Fe2+ b oxygen ha bi Thiobacillus ferrooxidans thnh Fe3+ + Fe3+ phn ng ha hc vi FeS2 thnh Fe2+ v H2SO4 + nhim mi trng xt v Fe2+ r r vo nc
Hnh 13.7. Hin tng nc r cha acid v chu trnh st 5.5. Lm giu qung bng vi sinh vt - ng dng trong lm giu qung khong ng, uranium) bng vi sinh vt: + Qung b x ha bi Fe3+ trong iu kin k kh thnh dng ion tan + Ion kim loai b kh thnh kim loai bi Fe to Fe2+ + Fe2+ b oxygen ha bi Thiobacillus ferrooxidans thnh Fe3+ Chuyn ha Hg - Hg c phng thch nhiu vo mi trng t cng nghip nng dc, in t, ha cht... - Cc dng lc kh khc nhau ca Hg: Hg0, Hg2+, CH3Hg+, CH3HgCH3, HgS, trong CH3Hg+ c c tnh cao nht v c tnh t trong chui thc phm
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Hnh 13.8. Chuyn ha Hg Cc chuyn ha: + Oxygen ha Hg0 thnh Hg2+ do phn ng quang ha + Methyl ha Hg2+ thnh CH3Hg+ v CH3HgCH3 do vi sinh vt + Sulfide ha Hg2+ thnh HgS bi vi khun kh sulfate + Kh CH3Hg+ thnh Hg0 v CH4 bi vi khun sinh methane 5.6. C ch khng c tnh thy ngn vi sinh vt - Pseudomonas aeruginosa: + Plasmid cha operon mer + Mt trong sn phm l Hg2+-reductase kh Hg2+ thnh Hg0 + Hg0 thng hoa khi t bo
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Hnh 13.9. C ch khng c tnh thy ngn vi sinh vt 5.7. Cc cht d sinh - Cht d sinh (xenobiotics) l cc hp cht do tng hp ha hc c phng thch vi s lng ln vo mi trng - Thuc tr su, thuc dit c
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Chuyn ha cc cht d sinh cha chlor - Kh chlor k kh (reductive dechlorination) cc thuc tr su cha chlor bi vi sinh vt : + Cht cho in t: Acetate, formate hay H2 + Cht nhn in t: thuc tr su cha chlor - Kh chlor hiu kh (aerobic dechlorination) bi vi sinh vt: sau bc kh chlor l bc phn hy vng benzene bng tc dng ca cc oxygenase cui cng thnh cc axt bo bc thp - Trong t nhin kh chlor k kh c vai tr quan trng hn do xu th hnh thnh mi trng khng c oxygen Cht do tng hp b phn hy sinh hc
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Hnh 13.10. Cht do tng hp b phn hy sinh hc
II. S PHN B CA VI SINH VT TRONG T NHIN

Vi sinh vt phn b v cng rng ri trong t nhin. Trn mt hoc nhiu khi bn trong tt c cc vt th sng hoc khng sng trong t nhin u c nhiu t vi sinh vt. Trong phn ny ta ch nghin cu v s phn b ca vi sinh vt trong khng kh, t v nc.
1. Vi sinh vt trong khng kh

Khng kh khng phi l mi trng thun li cho vi sinh vt pht trin. Tuy nhin trong khng kh vn c vi sinh vt do cun theo bi t v s h hp v bi tit ca ngi v ng vt. a s vi sinh vt trong khng kh l loi hoi sinh nh cu khun, trc khun c bo t, nm mc, nm men. Nhiu khi trong khng kh cng c vi sinh vt gy bnh nh trc khun lao (Mycobacterium tuberculosis), trc khun bch hu (Corynebacterium diphtheria), lin cu khun tan mu, t cu khun gy bnh, trc khun ho g, virus cm, si, quai b... t bnh nhn hay ngi lnh mang mm bnh tit ra. S lng vi sinh vt trong khng kh thay i ty a im, dn s, ma trong nm. Trn ni cao s lng t hn di thp. Ni c tuyt ph 1m3 khng kh c 4- 5 vi khun. Khng kh thnh th c nhiu vi sinh vt hn nng thn. Nhng ni ng dn c, nhng cng cng, rp ht, cu lc b...thng c nhiu vi khun, virus do ngi bi tit ra lm nhim bu khng kh trong mt thi gian nht nh. Nhng nh lp xp, m thp cng c nhiu vi sinh vt. Khng kh quanh bnh nhn c nhiu vi sinh vt hn ngi lnh. Mc d vi sinh vt trong khng kh khng nhiu lm, khng sinh sn v pht trin, nhng cng cn ht sc ch , v khng kh l phng tin truyn vi sinh vt t ch ny sang ch khc. Khng kh cng l ngun nhim bn thc phm v gy nn mt s bnh truyn nhim.
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Sau nhng trn ma ln, khng kh tr nn trong sch. Cc bin php trng cy xanh, xy dng nh ca cao ro thong kh, hng ngy m ca phng , phng lm vic cho khng kh lu thng, nh sng di vo u c tc dng tt trong vic lm sch khng kh. Trong mt s trng hp kh trng khng kh, ngi ta thng dng mt vi cht: acid lactic, formol, trietylenglycol, ozone, gn y ngi ta dng n pht tia t ngoi st trng khng kh cc phng m, phng sn xut v bo qun lnh trong cng nghip thc phm.
2. Vi sinh vt t
t l mi trng thun li cho vi sinh vt sng v pht trin (thc n, m, nhit pH...). S lng v kt cu khu h vi sinh vt thng xuyn thay i ty theo loi t, khu vc a l, tng t, thi v v ch canh tc. Thnh phn vi sinh vt t rt phc tp bao gm: vi khun, x khun, vi nm, vi to, ng vt nguyn sinh. Ring vi khun rt phong ph, bao gm vi khun hiu kh, k kh, t dng, d dng, vi khun c nh m... Trong t canh tc, c nhiu cht hu c, thong kh, s lng vi sinh vt ln ti hng triu c th/trong 1gam t, t hoang ha, t sa mc c s lng t hn. S lng vi sinh vt thay i theo su ca t: trn b mt c nh sng mt tri, kh ro nn c t vi sinh vt, su 0,5- 25cm, c s lng nhiu, 100- 200cm s lng bt u gim, su vi mt, vi sinh vt t tn ti v thiu cht hu c v oxy. Cc nhm vi sinh vt trong t thng xuyn c lin quan vi nhau, c khi tc ng tng h ln nhau nh Nitrosomonas v Nitrobacter, c khi chng i nhau nh cc loi vi khun, vi nm, x khun pht sinh ra cc cht khng sinh c ch s pht trin ca cc loi vi sinh vt khc. Vy h vi sinh t chnh l ngun ly nhim cho nc, khng kh. Ri chng theo nc v khng kh lan truyn khp mi ni. Cn phng vi sinh vt t lm h hng lng thc thc phm, v c th gy mt s bnh him ngho nh : bnh than, bnh un vn, hoi th sinh hi, ng c tht... Mt khc i vi vi sinh vt t c tm quan trng c bit trong s hnh thnh cht mn. Trong t cn c nhng vi khun c nh N2 sng t do hoc cng sinh lm giu thm nit cho t. Nhng ngc li trong t cng c nhng vi khun phn nitrat ha, phn sunfat ha..., mc ph nhiu ca t ph thuc vo s hot ng ca chng. Do ta phi khng ch hot ng ca chng pht huy nh hng tt, h thp nh hng xu, a nng sut ma mng ln cao. t cng l ngun tt phn lp VSV hu ch.
3. Vi sinh vt nc
Nc l mt vn quan trng trong i sng cng nh trong cng nghip thc phm, dc phm... hiu r vn ny ta tm hiu s phn b vi sinh vt trong nc, vn lm sch nc v vn nc thi.
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Cng nh t, nc t nhin l mt mi trng rt thun li cho vi sinh vt tn ti v pht trin. S pht trin ca vi sinh vt trong nc ph thuc vo mt s yu t nh nhit , pH, nng O2..., trong quan trng hn c l thc n trong nc. Nc cng b bn do cc b hu c- ngha l cng nhiu thc n- th cng cha nhiu vi sinh vt. iu ny ta s thy r khi nghin cu tng loi nc. -Nc ngm (nc mch) c t vi sinh vt l do nc c thm qua cc lp t dy, vi sinh vt v cc thc n hu c c gi li trong lp t ny. Nc ngm cng su th cng sch. Cc ging mch nu tm c ngun mch tt, su v c bo v trnh nhim bn b th s cho ta nc rt tt cho vic n ung. - Nc ao, h trng hp b nhim phn, rc ri c nhiu cht hu c th s lng chng loi vi sinh vt tng nhiu. - Nc trong nhng h, bin ln bi b lng chm nn s lng vi sinh vt t hn. - Nc nhng vng sng, ngi gn dn c th s lng vi sinh vt nhiu hn vng xa dn c. Nc trong t nhin c kh nng t lm sch do tc dng ca nh sng mt tri v do s cnh tranh sinh tn m hy dit ln nhau, hoc l do cht khng sinh ca cc thc vt thy sinh tit ra. Nc cng l ngun truyn bnh nguy him nh vi khun thng hn (Salmonella typhi, trc khun l (Shigella spp.), phy khun t (Vibrio cholerae)..., ngoi nhng vi sinh vt sng trong nc, cn c nhng vi sinh vt gy bnh do ngi v ng vt lm nhim. Cc vi sinh vt gy bnh ny ch tn ti trong nc mt thi gian nht nh. Chng tn ti trong nc lu hay mau ty theo ngun nc, tnh cht, nhit , pH... ca nc.
III. MT S BIN PHP VI SINH GP PHN BO V MI TRNG

1. Nguyn nhn gy nhim mi trng
1.1. Ph rng: C th xem ph rng l nguyn nhn c bn, l ngun gc ca nhiu nguyn nhn khc lm mt cn bng h sinh thi v gy nhim mi trng. 1.2. S pht trin cc ngnh cng nghip: L nguyn nhn gy nhim mi trng do cc nh my thi ra cc cht thi dng kh, dng lng hay dng rn, u l nhng cht gy nhim mi trng. 1.3. Thuc tr su, phn ha hc: Cng l nguyn nhn quan trng gy nhim mi trng khi s dng qu nhiu, s dng ba bi thiu cn i. 1.4. Hot ng ca con ngi: Gp phn lm nhim mi trng bi cht thi sinh hot, bi cc hot ng sn xut v i sng ca con ngi.
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Ngoi ra cn nhiu nguyn nhn khc cng gy nhim mi trng, c nguyn nhn thng xuyn, c nguyn nhn xy ra t xut. V d: chy rng, m tu ch du, tai nn nh my ht nhn, th v kh ht nhn, v kh ha hc, chin tranh
2. Mt s bin php vi sinh gp phn bo v mi trng

Vic bo v mi trng c th thc hin theo cc phng php c bn l: - Trit tiu nguyn nhn hay hn ch nguyn nhn gy nhim mi trng. - X l hu qu m cc nguyn nhn gy ra s nhim. Vi nhng phng hng , CNVS c th gp phn bo v mi trng theo cc bin php sau: 2.1. Trng cy xanh: Trng cy xanh ph xanh t trng i trc, ci to t, ci to mi sinh, ci to h sinh thi. . . l bin php hu hiu hn ch s nhim mi trng do rng b tn ph gy ra. 2.2. Dng phn sinh hc thay phn ha hc Hn ch s dng phn ha hc nh CNSH to ra cc dng phn sinh hc nh Nitragin, azotobacterin, phn ln vi sinh. . . Nh gim thiu s nhim do phn hc hc gy ra. 2.3. Dng phng php sinh hc bo v thc vt thay cho thuc tr su - Dng VSV dit cn trng gy hi - Dng ng vt dit cn trng gy hi - Cng ngh gen to ging cy chu su bnh 2.4. X l cht thi bng CNVS
3. Mt s bin php hin ang c p dng trong nc v trn th gii

3.1. Cng ngh c nh nitrogen v sn xut phn bn vi sinh 3.1.1. Cng ngh gen to ging cy c kh nng c nh m C 2 hng gii quyt vn ny ang c thc hin: - S dng cng ngh gen: Chuyn gen sn xut nitrogenaza (gen nif) t vi khun Klebsiella pneumoniae c kh nng c nh nit sang E.coli to ra cho E.coli c kh nng c nh m ri s dng chng nh l ngun cung cp m. - Chuyn gen nif vo cc cy khng c kh nng c nh m lm cho cc cy ny tr thnh cy c kh nng c inh m nn t cung cp c m cho cy. Hy vng rng trong thi gian n, bng k thut gen ngi ta chuyn cc plasmid cha gen nif vo cc cy trng v s chm dt nn i m trin min ca cy trng, lc by gi chc chn s to ra mt bc ngot v i i vi s pht trin nng nghip trn ton th gii. 3.1.2. Cng ngh sinh hc vi sn xut phn vi sinh vt
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Hin nay ngi ta to ra nhng ging cy trng c nng sut cao thng nhu cu i hi v phn bn cng cao. V vy m t khng p ng c nhu cu cho cy nn phi s dng nhiu dng phn bn: Phn v c, phn hu c, phn vi sinh... Phn v c hay cn gi l phn ha hc c u im nh cho kt qu nhanh, hiu qu cao, nhng nu dng nhiu s lm nhim mi trng, lm thay i tnh cht l ha ca t theo hng lm cho t xu i, tch ly nhiu ion c trong t v ngay c trong cc sn phm thu hoch. . . Phn hu c c nhiu loi nh phn xanh, phn bn, phn rc, phn chung. . . nhng hiu qu chm, hiu sut kinh t khng cao do phi s dng lng ln, tn km cng vn chuyn. . . Phn vi sinh c nhiu dng v c hiu qu khc nhau ph hp vi loi cy trng cng nh loi t trng. Ph bin hin nay l phn vi khun nt sn (Nitragin), Azotobacterin, phn ln vi sinh. 3.1.3. Phn vi khun nt sn (Nitragin): Ch phm vi khun nt sn (c tn gi nitragin) ny c sn xut v s dng rng ri M, H Lan, B, Php. . . Ty tng nc m nitragin sn xut ra di nhng hnh thc khc nhau: trn thch, trong dch th, hp th vo than bn, t vn. . .
[1] [2] Hnh 13.11: Ch phm Nitragin [1] v Hiu qu ca phn bn nitragin [2] Qu trnh sn xut nitragin c tin hnh theo cc bc sau: - Phn lp vi khun nt sn t nt r cc cy h u - Tuyn chn cc chng c hot tnh c nh m cao, c kh nng chu ng c cc iu kin bt li ca mi trng v c kh nng cnh tranh c vi cc VSV khc sng xung quanh r cy; c kh nng sng st trn ht khi tm nhim. - Nhn ging ri cho hp th vo cht mang. Theo GS.Nguyn Ln Dng trong hon cnh nc ta hin nay trc ma gieo mt loi u no , cc phng th nghim nh t chc cc tnh s sn xut cc ng nghim thch nghing cy vi khun nt sn v chuyn trc tip xung cc hp tc x, cc nng
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trng, y ging c nhn ln trong cc mi trng n gin (cha nc chit u 5%, vi 1% ng knh). Sau 72 gi nui cy trong mi ml mi trng ny c th t ti 3800.1016 vi khun nt sn in thanh hoc 2800.1016 t bo vi khun nt sn lc. em tm vo ht hoc bn vo t. . . Cn ch trnh s dng ng thi cc loi thuc tr su, thuc dit c c kh nng tiu dit hoc c ch s pht trin ca vi khun nt sn. 3.1.4. Phn Azotobacterin Trong nhiu nm nhiu nc khc nhau, ngi ta sn xut quy m cng nghip hay th cng nhng ch phm t vi khun Azotobacter v gi l Azotobacterin. l nhng dch nui cy Azotobacter c hp th vo than bn hoc cc loi t giu cht hu c trung ha v b sung thm mt t phn photpho kali. . . Azotobacter l loi vi khun sng t do hiu kh, t bo t hnh cu n hnh que. Khi cn non t bo hnh que kch thc khong 2,0-0,7 x 1,0-2,5 m di ng nh chu mao, khi tiu th ht 1g cc hp cht carbon c kh nng c nh c 10mg N 2. pH thch hp 7,28,2, ngoi kh nng c nh m Azotobacter cn c kh nng to ra lng ln cc cht iu ha sinh trng nh auxin, gibberellin. . . 3.1.5. Phn ln sinh hc Trong nhiu nm gn y, tip theo vic nghin cu v s dng phn m sinh hc thay cho phn m ha hc ngi ta cng i su nghin cu vic ch to phn ln sinh hc. phn hy dng ln hu c kh tan, ngi ta s dng nhm VSV c kh nng phn hy ln hu c nh: Bacillus mycoides, B.subtilis, Proteus vulgaris . . . phn hy ln v c, cn s dng VSV nhm: Bacillus megatherium. B.mycoides, B.butyricus, Pseudomonas gracilis, P.herbicola, P.radiobacter. . . Nm mc Aspergillus niger. Qu trnh sn xut phn ln sinh hc tin hnh qua cc bc ch yu sau: - Phn lp v thun khit cc VSV c kh nng phn gii ln - Tuyn chn chng c hot tnh phn gii ln cao, c kh nng thch hp vi cc iu kin sng khc nhau trong mi trng t, c kh nng tn ti trong cht mang khi sn xut ch phm. - Nhn ging VSV gia tng s lng VSV phn gii ln - Ch to ch phm bng cch trn VSV phn gii ln vi cht mang vi t l thch hp. Cht mang thng dng l than bn, t bt apatit, c th b sung thm 1% ng lm cht dinh dng cho vi khun pht trin tt, ch phm c th dng bt hay dng vin. 3.2. Thuc sinh hc bo v thc vt Vic s dng qu nhiu thuc ho hc tr su v dit c di trong nng nghp ngy cng dn n hu qu nhim mi trng v ph hoi h sinh thi t nhin. Thuc sinh hc bo v thc vt da trn quan h i khng gia cc nhm vi sinh vt vi nhau, gia cn trng vi nhau, cng nh gia VSV vi cn trng. Cc quan h i khng ny vn c trong t nhin, n chnh l mt trong cc yu t t iu chnh ca h sinh thi. Khi mt loi no trong h sinh thi pht trin qu nhiu v s lng th c mt loi khc dng n lm
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thc n cng pht trin mnh v tiu dit n. Nh cc quan h i khng nh vy, cc loi trong t nhin lun lun c gi th cn bng. H sinh thi t nhin c bo tn nh nhng quy lut t iu chnh nh vy. Khi s dng qu nhiu thuc tr su v dit c, cc loi i khng vn t li b tiu dit i cng vi cn trng gy hi, v v th yu t iu chnh t nhin ngy cng mt i. Phng php sinh hc bo v thc vt nhm mc ch phc hi nhng loi i khng vi cn trng gy hi, da trn nguyn tc nui nhn to cc loi i khng, th ra thin nhin nhng ni c nhiu loi gy hi. C th s dng bin php cn trng dit cn trng nh nui nhn to ong mt Tricoderma dit tr su xm v mt s loi su khc. C th s dng bin php dit cn trng nh nh nui cy Bacillus thuringiensis gy bnh cho cc loi su hi cy trng. Trong t bo ca loi vi khun B. thuringiensis c nhng ht tinh th hnh qu trm c c tnh i vi cn trng. Nhng vi khun dit cn trng u tin c phn lp ra u tin t cn trng b bnh. Sau c nui cy, nhn ging v cui cng c a tr li thin nhin, gy dch bnh hng lot cho cn trng hi cy. Hin nay c hai nhm bin php c th s dng VSV phng tr bnh cy: dng VSV dit cn trng v dng VSV dit VSV bng kh nng sinh cht khng sinh. 3.2.1. Bin php dng VSV dit cn trng. Bin php VSV dit cn trng da trn c s: c nhiu loi VSV c kh nng gy bnh cho cn trng hi cy. Nhng loi ny ch yu nm trong 3 nhm chnh: vi khun, vi nm v virus. t hu c ch phm dit cn trng phi qua cc bc sau: * Phn lp vi khun B. thuringiensis(BT) : - phn lp vi khun BT ngi ta thu mu l nhng cn trng b bnh ang cn sng. - S dng mt trong cc mi trng c thnh phn nh sau Bng 13.1 Mi trng dng phn lp vi khun B. thuringiensis Nguyn liu (g/l) Mi trng 1 Mi trng 2 Mi trng 3 Mi trng 4 Glucose 15 5 3 Saccaroza 10 Cao ng 4.5 Tinh bt ng 6.0 5 Cao nm men 3.5 2 K2HPO4 1 KH2PO4 1 1 NaOH 0.43 CaCl2 0.1 Pepton 10 30 10 NaCl 2 5 MgSO4.7H2O 0.1 Na2HPO4 5 Cao tht 3 (Dn theo Trn Cm Vn, 1995).
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* Tuyn chn chng thch hp: Cn tuyn chn chng vi khun BT mang cc c im: - C hiu lc dit su cao - C t l sng st cao khi phun mt s lng ln ra ngoi ng rung - C kh nng sinh trng, pht trin, gi hot tnh trong iu kin nui cy nhn ging phng th nghim. - C t l sng st cao trong ch phm. *Ln men to ch phm: sn xut ch phm BT c 3 phng php chnh: Ln men trn mi trng lng (ln men chm), ln men trn mi trng c v ln men trn mi trng bn lng (cn gi l ln men xp). Sau dy chng ti xin gii thiu phng php ln men chm: - Nhn ging cp 1; Mi trng c thnh phn nh MT 1 Bng IV.1 - Nhn ging cp 2: Mi trng c thnh phn (g/l): Glucose -3; Kh lc - 5; Bt c5; Cao ng-5; Du u tng- 2.5ml; nc 1lit. - Ln men : Mi trng ln men chm c thnh phn g/l): Kh lc - 20; Bt c- 5; Cao ng-18; CaCO3- 5; MgSO4.7H2O-0.07; Dextrin- 5; K2HPO4- 0.4; Du u tng- 3ml; nc 1lit. Thnh phm: Ch phm sau khi ln men phi nh gi cht lng da vo 2 ch tiu: s lng vi khun sng trong ch phm t trn 2.109 t bo/g v c tnh ca ch phm phi mnh.
Hnh 13.12 Ch phm Thuc tr su BT v tinh th hnh qu trm ca vi khun B. Thuringienssis c c tnh i vi cn trng. b.Bin php VSV dit VSV gy bnh bng kh nng sinh cht khng sinh. Cy trng khng nhng b ph hoi bi cn trng m cn b thit hi do cc nhm VSV gy bnh cy. Nc ta nm trong vng nhit i nng m, rt thch hp cho cc loi VSV gy bnh cy, c bit l nhm vi nm. V d nh bnh o n do nm mc Piricularia oryzae gy ra, bnh tim la do Heminthosporum oryzae, bnh la von do Fusarium moniliformae gy ra. Cc bnh ny gy thit hi ng k cho sn lng ma mng. Thuc ho hc dng tiu dit cc loi nm bnh rt nhiu, nhng dng cng nhiu liu lng ngy
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cng phi cao ln do hin tng nhn thuc. Liu lng cao ca cc cht ho hc tiu dit c khu h VSV c ch trong t nh VSV c nh nit , VSV phn hu cht hu c..... Bin php s dng VSV dit VSV gy bnh cy bng kh nng sinh cht khng sinh da trn hai mi quan h c bn gia VSV vi nhau: Quan h k sinh v quan h i khng. Quan h k sinh: VSV gy bnh cy sng k sinh trn thc vt, nhng chnh n li b mt loi VSV khc sng k sinh- gi l k sinh bc hai. K sinh bc hai thng l vi khun hay virus c kh nng sng k trn vi nm gy bnh thc vt. Nguyn tc ca bin php ny l tch cc k sinh bc 2 t nm bnh thc vt, nui cy, nhn ging trong iu kin th nghim ri th tr li ti vng c k ch ca n ang pht trin. K sinh bc hai s gy thnh dch bnh lm cht hng lot cc VSV gy bnh cho cy trng. Kh khn ca bin php ny l vic phn lp cng nh nui cy k sinh bc hai. V n l loi sinh vt sng k sinh trn c th sng nn vic phn lp v nui cy bng mi trng nhn to rt kh khn. Quan h i khng: VSV trong qu trnh sng tit ra cc cht i khng c ch hoc tiu dit ln nhau. in hnh l cc VSV sinh cht khng sinh. Vic nghin cu v ng dng bin php ny c tin hnh theo cc bc: -Phn lp chng VSV sinh khng sinh -Chn chng, nng cao hot tnh. -Th hot tnh khng sinh i vi VSV gy bnh cy - Sn xut v th tc dng cc ch phm. Bn cnh vic nhp ging VSV sinh cht khng sinh c hot tnh mnh t nc ngoi, vic phn lp cc chng VSV c kh nng sinh cht khng sinh l rt quan trng v cht khng sinh thuc loi hp cht d b phn hu trong mi trng t, nht l iu kin nng rn. Cc VSV c ngun gc nc ngoi thng khng thch hp vi iu kin t nhin ca nc ta, khi a vo t d b mt hot tnh. Trong khi khu h VSV t ca Vit Nam rt nhiu v s lng v phong ph v chng loi. * To chng VSV sinh cht khng sinh: Trong s cc VSV sinh cht khng sinh ng ch l x khun. phn lp x khun sinh khng sinh thuc chi Streptomyces cn s dng mt trong cc mi trng c thnh phn nh (Bng 13.2): c ch cc loi tp khun, nn b sung vo mi trng phn lp x khun K2Cr2O7 vi t l 0,4g/l. th hot tnh khng sinh ngi ta thng dng phng php khuych tn trn thch. Cc chng x khun dng trong bo v thc vt cn c chn theo nhng tiu chun sau: - Hot lc khng sinh cao - Hot ph khng sinh rng
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- Gi c hot tnh khi a vo ng rung - Khng c i vi cy trng, ngi, ng vt v cc VSV khc. Khng thuc loi khng sinh dng trong y hc. Bng 13.2. Thnh phn mi trng phn lp x khun chi Streptomyces. 1. MT ISP- 4(g/l) : 2. MT Gause- 1(g/l) : Tinh bt tan 10 thnh phn dung dch Tinh bt tan 20 vi lng : K2HPO4.3H2O 0,5 K2HPO4.3H2O 1 FeSO4.7H2O 0,1 MgSO4.7H2O 0,5 MgSO4.7H2O 1 ZnCl2 0,1 NaCl 0,5 NaCl 1 SnSO4 0,1 FeSO4.7H2O 0,01 (NH4)2SO4 2 H2O 100ml KNO3 1 CaCO3 2 Thch 20 Dung dch vi lng1ml Nc ct 1lit Thch 20 pH 7,2- 7,4 Nc ct 1litt pH 7,0 * To ch phm khng sinh th: Hin nay, cc ch phm khng sinh dng trong nng nghip c sn xut dng th. C hai dng ch phm: dng dch th v dng bt. Ch phm dng bt tin li cho qu trnh bo qun v vn chuyn hn ch phm dng nc. Khi s dng, tu theo i tng bnh cy Dng dch th: Ging x khun c cy truyn sang mi trng thch nghing (MT-Gauze1). Tin hnh nhn ging cp 1 trn mi trng c thnh phn (g/l): Bt ng-20; (NH4)2SO4 -5; MgSO4.7H2O-2; K2HPO4-2; NaCl- 2; nc 1lit. Phn phi mi trng vo cc bnh tam gic dung tch 500ml, mi bnh ng 100ml. Mi bnh cy mt ng ging x khun thch nghing (ht 1-2ml mi trng lng cho vo ng ging, dng que cy gt cho bo t ho vo dung dch. dung dch tr li bnh tam gic. Nui cy trn my lc tc 200- 220vng/pht nhit 28- 300C, trong thi gian t 24- 48 gi. Nhn ging cp 2 vi mi trng c thnh phn (g/l): Bt ng- 30; (NH4)2SO4- 4; CaCO3- 4; NaCl- 2; nc 1lit; pH= 6.5- 7.5. Mi trng ln men: c th s dng mt trong cc mi trng sau y (g/l): 1).MT1: Cao ng- 0.5; Glucose- 1; CaCO3- 0.5; NaCl- 0.5; (NH4)2SO4-0.35; Bt khoai ty - 1.5. 2).MT2: Cao ng- 0.3; Bt u tng- 2; Glucose- 3; CaCO3- 0.6; NaCl- 0.2; (NH4)2SO4-0.3; KH2PO4- 0.02. 3).MT3: Bt ng- 2; Nm men kh- 0.5; Glucose- 3; CaCO3- 0.6; NaCl- 0.2; (NH4)2SO4-0.3; KH2PO4- 0.02. Dch nui cy x khun sau khi kt thc giai on nui trong ni ln men c sc kh c th coi nh ch phm khng sinh th. Ch phm ny c th em ly tm loi b sinh khi x
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khun ri em ng chai bo qun nhit thp (nu phun ln l). Cng c th c khi x khun nu mun a x khun vo t. *ng dng ch phm khng sinh th: - Bn trc tip vo t, vo gc cy - Phun dch khng sinh th ln l, ln thn nn loi b sinh khi x khun thay vo l cc cht gip cho thuc bm dnh trn thn l. 3.3. Cng ngh x l rc thi hu c Rc thi l mt nguyn nhn quan trng gy nhim mi trng. Mc sng ca nhn loi ngy cng cao, dn s cng tng th mc nhim mi trng do rc to ra cng ln. Thnh phn rc thi rt a dng, cc rc thi hng ngy c thu gom li v x l bng nhiu phng php khc nhau. Trc y, phn ln rc thi thnh ph, cc khu ng dn c c tp trung li em chn lp hay t i, c hai bin php ny u gy nhim mi trng. - Rc chn xung t s lm nhim ngun nc, nhim vng t xung quanh bi rc thi, cc kh thi c hi do qu trnh phn hy t nhin bay ra lm nhim bu khng kh. - Rc em t s thi vo bu kh quyn mt lng ln cc kh c. khc phc cc nhc im trn, ngi ta dng bin php sinh hc x l rc c hiu qu nht: va khng gy nhim mi trng, li va c hiu qu kinh t cao. 3.3.1. X l rc thi bo v mi trng v tn thu lm phn bn GS.TS. Phm Vn Ty cng mt s nh khoa hc khc kt hp vi cng ty mi trng v th H Ni nghin cu a VSV c kh nng phn gii xelluloza vo x l rc thi. Theo phng php ny, rc c nh thnh ng hnh thang, 2m x 3m x 1,5m, di y ng c ng ng dn kh vo. Bo t nm mc c hot tnh phn gii xelluloza cao c b sung vo theo t l 40-50g/tn rc. Dng my thi kh vo trong sut tun u mi ln 10 pht, mi ln cch nhau 10 pht. Trong tun 2 v 3 mi ln cch nhau 10 pht v mi ln ko di 5 pht. Tun 4 ngng thi kh, sau 40 ngy kim tra, nu rc phn hy tt em sng cn nu cha c th i thm vi ngy. Phng php ny c th phn hy khong 30% rc hu c sau 40-50 ngy. Theo n VIE/86/023 nh my th im x l rc Cu Din c ci to v xy dng theo cng ngh x l rc thi bng phng php ln men, s dng h thng qut cung cp kh cng bc to iu kin rc phn hy thun li. Nhit khi khong 500C v trong trng hp cn thit kt hp o trn n nh nhit , thi gian x l thng thng khong 60 ngy. tng thm thnh phn nit trong phn trn thm ( cn thi bm ht cc nh v sinh t hoi) trong qu trnh ln men, dy chuyn cng ngh nh sau (Hnh 13.13) n quy trnh cng ngh dng VSV phn gii hiu kh rc thi th v rc thi nng nghip nh sau: Rc c cht thnh ng ln men, c sau 10 ngy ng rc li
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c o trn mt ln. Trong thi gian 33 ngy, ng rc c phn hy hon ton v thu c phn hu c c tn gi l CELRICH. CELRICH c mt s u im nh: hn ch c di mc, c cha VSV phn gii ln v VSV c nh m, nng cao nng sut ma mng 1540%; ci thin mu sc, mi v kch thc hoa qu v rau trong iu kin tn t phn bn ha hc.
Hnh 13.10. S cng ngh x l rc thi ti x nghip x l rc thi Cu Din 3.3.2. X l nc thi bng cng ngh sinh hc Nc c dng nhiu trong sinh hot, trong sn xut nng, cng nghip. Nc sau khi s dng c thi ra mi trng. Nc thi cha nhiu cht v c, hu c, nhiu kh c, c bit nc thi cc bnh vin cha nhiu loi VSV gy bnh. Do vic x l nc thi, bin nc nhim bn thnh nc sch tr li cho mi trng, hay s dng li cho sn xut v i sng l vic lm cp thit. Hin nay trn th gii ang s dng cc phng php vt l, ha hc v sinh hc x l nc thi. Trong cc phng php trn, phng php sinh hc ngy cng c s dng rng ri v nhng u im: - Phn hy hot cht hu c trong nc thi nhanh, trit m khng gy nhim mi trng. - To ra c mt s sn phm c ch s dng trong cng nghip v sinh hot nh: bioga, etanol..., trong nng nghip nh phn bn. - Thit b n gin, phng php d lm, chi ph t tn km, hiu qu cao hn cc phng php khc. Phng php sinh hc x l nc thi thng c th bao gm cc gii php chnh l:
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Phng php x l hiu kh ( x l bng bn hot tnh): y l phng php ph bin nht cho x l nc thi hu c, trong phng php ny, cc cht hu c nng cao c trong nc thi c VSV s dng trong iu kin hiu kh. Sinh khi VSV c nui trong thit b sc kh, c th bng kim loi hoc b tng v c lng trong thit b lng tip sau, nc thi sau khi x l c tch khi cn bn nh thit b lng ny. tng cng hiu qu ca b hiu kh v ca VSV, sau khi tch khi nc trong b lng, lng bn nc c hi lu mt phn v b hiu kh. Phng php ny s dng cho cc nc thi c hm lng BOD trung bnh t 150-10.000mg/l. H vi sinh vt: thng gp l: Actinomycetes, Bacillus, bacterrium, Pseudomonas... Phng php mng sinh hc: Trong phng php ny, VSV c c nh trn b mt ca cht mng, nc thi c a vo trn lp vt liu mng. Ti y, cht hu c c hp th v phn gii. Qu trnh phn gii cc HCHC ch yu nh s c mt ca cc VSV hiu kh trn b mt mng v v vy, chng c phn gii ti CO2 v H2O. Ngoi ra trong cc lp su ca mng c cha cc vi khun ym kh, ng vai tr phn gii cc HCHC thnh H2S, NH+4, axit hu c... cc hp cht ny li tip tc c vi khun hiu kh phn hy ti H2SO4, HNO3, CO2 v H2O. Ngoi vi khun, s phn hy cc HCHC cn c s ng gp ca cc loi vi to v cc ng vt nguyn sinh khc. Phng php mng sinh hc nhn chung thng c p dng x l cc nc thi c nhim bn thp, BOD t 100-200mg/l vi hiu sut x l 75-90%BOD. Phng php h sinh hc: Phng php x l kt hp tc dng ca vi khun vi cc sinh vt khc, ch yu l to v cc VNS khc, ng thi kt hp hot ng ca vi khun hiu kh, ym kh nh to ra mc thong kh khc nhau trong h c chiu su khng qu ln, kt hp vi hiu qu chiu sng ca din tch b mt h. Kh nng phn gii khong 35-95% cc hp cht hu c. Phng php x l ym kh: Phng php ny thch hp cho x l cc nc thi c nng nhim bn cao, BOD t 500mg/l hoc ln hn v c kh nng thu hi nng lng, to ra t bn kh nng phn hy ti 80-90% BOD. Trong cch x l ym kh, vi khun ym kh c nui trong cc thit b ym kh c khuy trn. Cht hu c c phn gii ti metan, CO2, cc axit hu c, H2S... Mt lng ln nng lng thu c di dng kh metan. Qu trnh phn hy ym kh cc HCHC c th xy ra nhit cao 53-550C nh h vi khun Themophillus (tc phn hy trong iu kin trn cao hn 2-3 ln so vi tc phn hy nh h mesophillus 34-360C. Phng php t nhin x l rc thi: Vi nhng loi rc thi c nng cht nhim bn khng cao, khng cha nhiu cht c hi cho ngi th thng dng phng php x l t nhin. Nc thi t nh my, khu dn c... cho chy thm qua lp t dy khong 2m, hay cho chy qua lp t xp, ct, si... nc thi i qua cc lp t nh h VSV c sn y cc cht bn s c phn hy lm cho nc tr nn sch hn. y l qu trnh t lm sch
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d lm, t tn km, c th x l nc thi trn quy m ln. Nc sau khi x l c th dng trong nng nghip, dn ra ao h sng... ha vo ngun nc chung ca t nhin.
Cu hi n tp chng 13
Chn p n ng nht cho mi cu hi 1. S phn nitrate ha c lin quan ti:
-
S phn nitrate ha ammoniac thnh N2 S chuyn N2 thnh ammoniac S chuyn nitrite thnh ammoniac S chuyn nitrate thnh nitrite Khng c no trong cc trn M hnh ha mt h sinh thi nc nh mt h nc ngt Sn xut cc enzym cho cc ng dng cng nghip Loi cc ha cht c khi ngun nc Liu php sinh hc t Lng carbon tng s trong h thng Lng oxi b tiu th bi mt h thng nc Lng oxi trong mt h thng nc nhn c cho s s dng sinh hc Lng carbon c th chuyn ha d dng trong mt h thng nc Tt c cc trn
2.Chc nng ca ct Winogradski l g ?
3. BOD l g ?
Chn mt cu tr li ng nht cho mi cu hi 1. .l nhng vi khun lc ca cc t bo vi khun c gn ln mt b mt nh cc polisaccarid dnh do t bo tit ra Qun x FISHs Vi mi trng Mng sinh hc Xc nh qun th vi sinh vt ca rut ngi Xc nh cc trnh t du chun trong mt mi trng t M hnh ha s sn xut methane trong mt ng vt nhai li M hnh ha cc mi trng t v nc nh l cc mi trng tn ti y ca mt ci ao.
2.ng dng no sau y l mt s ng dng in hnh ca ct Winogradski ?
3. Nguyn nhn no sau y s l mt nguyn nhn quan st thy rng VSV pht trin trong t nhin chm hn trong nui cy phng th nghim
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S phn b cht dinh dng t khi ng u khp mt mi trng Cc cht dinh dng thng c cung cp nng thp trong t nhin Vi sinh vt trong t nhin t khi tn ti trong ging thun khit v phi cnh tranh v cc ti nguyn. Tt c iu trn u l nguyn nhn
Tr li cc cu hi sau: 4. Nguyn nhn gy nhim mi trng ? nhng bin php no ca cng ngh vi sinh gp phn chng nhim mi trng ? 5.Cc loi phn bn vi sinh vt ? Trnh by quy trnh sn xut mt loi phn bn vi sinh thng dng hin nay. Cho v d minh ho. 6.Trnh by c s khoa hc ca mi quan h i khng gia cc sinh vt. Qua hy cho bit quy trnh sn xut v ng dng ca mt loi thuc tr su sinh hc. 7. C s khoa hc ca cc bin php x l rc thi hu c ? Trnh by mt quy trnh c th.
* Ti liu c thm
1. Kiu Hu nh, 2006. Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni, 2. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006. Vi sinh vt hc; NXB i hc Hu. 3. Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty. Vi sinh vt hc, 1997, NXBGD H Ni.
* Ti liu tham kho

* Gii thch thut ng

Nc acid thot t khai thc qung : nc acid cha H2SO4, bt ngun t s i ha sunfua st nh vi sinh vt. Mng sinh hc : cc khun lc ca cc t bo VSV c bao bc bi mng nhy v c gn vo mt b mt. Sinh a ha hc : S nghin cu v nhng chuyn ha ha hc xy ra theo con ng sinh hc. FISH(fluorescent in situ hybridization) :S lai hunh quang in situ In situ: trong mi trng Vi mi trng: mi trng trc tip bao quanh mt hay mt nhm t bo vi sinh vt.
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CHNG 14
CU HI N TP I. PHN CU HI
1. in vo ch trng Cu 1: a. Th gii vi sinh vt v cng a dng. nghin cu chng cc nh khoa hc phi da vo cc tiu ch v ...................,........................,......................... sp xp chng vo bc thang phn loi v t tn. b. Cc vi sinh vt c sp xp vo bc thang phn loi t thp n cao:....................................................................................Loi l bc thang phn loi thp nht.........l bc thang phn loi cao nht. Loi c t tn theo h thng..........................theo ting.....................vit nghing. V d vi khun gy bnh than c tn l Bacillus anthracis. Cu 2: Virus l nhng thc th gy nhim.................... m gen nom ca chng ch cha hoc l................ hoc l ............... . ....Chng ch nhn ln trong ........................... bng cch s dng b my...........................v .............................ca t bo tng hp nn cc bn sao ca bn thn mnh nhm truyn .................. ca bn thn chng sang cc t bo khc. Cu 3: Cu trc in hnh ca mt virus n gin nht bao gm mt............... (..............hoc..... ....) v mt .................c gi l ............... Cc virus phc tp hn, c bit l cc virus ng vt cn cha mt lp v bc c to nn bi mt t hp....................,.................v............................ Cu 4: a. DNA si n ca virus va c transcriptase ngc tng hp nn trc ht c chuyn thnh...... ..............v sau c.................................ca t bo ch. b. mt s virus, lp v bc c ph bi cc................ c bn cht l.................hoc..............m vai tr ca chng l nhn bit (v mt ho hc) cc ........ v gn vo t bo m chng gy nhim. Cu 5: a. La tui b nhim HIV cao nht l la tui ......................... b. Bnh vim gan B c truyn ch yu qua ng............ c. Bacteriophage nhn ln nh.................v ph hu n. l qu trnh............, cn cc vi khun cha mt prophage v lu gi n vnh vin trong...........ca mnh c gi l cc vi khun.............. Cu 6: a. Prophage, nm trn .....................ca vi khun, chnh l nguyn liu di truyn ca............................
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b. Cc ht tng t virus song ch cha ARN thun khit c gi l ..........l bn gy bnh thc vt c ngha kinh t nh............................ c. Cc ht protein thun khit gy bnh thn kinh ng vt c tn l........ Cu 7: a. Hin tng mt loi virus ch nhim bnh trn mt vt ch hay nhm vt ch nht nh c gi l .............ca virus. Tnh ny do cc.............................to nn. b. Bnh di l mt bnh ...................cp tnh thng dn n Cu 8: Trong cc cu trc sau y, hai cu trc no khng gp trong mt ht HIV: a) ARN si n; b) cc glicoprotein b mt; c) DNA si kp; transcriptose ngc; e) protein v; f) DNA si n. Cu 9: a. Nhim sc th vi khun khi nm di dng...................... s khc vi dng ................bi mt cu trc m c hn v t cng knh hn. b. Khi c mt ca lisozim v saccarose, mt trc khun Gram m s b bin i thnh mt t bo......................c tn l........................v mt i dng hnh que ca n. Cu 10: Nm l sinh vt thuc dng t bo ............................C th c th n hay a bo dng si, c thnh.......................(mt s t c thnh..................), khng c...............sng d dng theo kiu................. Sinh sn bng .............khng c...............v ............................................... Cu 11: S khc bit c bn ca cc t bo Prokaryote so vi t bo Eukaryote l ch: a. Nguyn liu di truyn ca chng ..... khng c bao bc bi........ b. Thiu cc bo quan nh:....................c bao bc bi mng.. c. Thnh t bo lun lun cha phc h..............hay cn gi l.......... d. Ribosome ca cc t bo prokaryote thuc loi ............................ Cu 12: a. Ni bo t l th ............... ca cc t bo vi khun, khng phi l phng tin sinh sn, khi gp iu kin thun li, bo t s ny mm v mi bo t chi cho ........t bo dinh dng. b.Ni bo t c tnh ........cao, do vy thng gy kh khn trong cng tc bo qun thc phm v trong vic kh trng cc bnh vin. Cu 13: a. Nhn t di truyn ngoi nhn ca vi khun c tn l cc ............., chng khng quyt nh s sng cn ca t bo vi khun song mang mt s gen quan trng nh cc gen.........................., ................................
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b. Cc vi khun lu hunh mu ta v mu lc s dng.............lm cht cho in t trong quang hp, kt qu l to thnh.......................tch lu bn trong t bo hoc c tit ra bn ngoi t bo. Cu 14: a. C nh nitrogen l qu trnh chuyn ho nitrogen ca kh quyn (N2) thnh........... mt cht cn thit cho s sinh tng hp cc amino acid. b. Trong th gii vi sinh vt, ch c mt s loi.................... l c kh nng c nh nitrogen phn t. c. Enzym chu trch nhim i vi s c nh nitrogen phn t c tn l .... d. C......nhm vi khun c nh nitrogen ch yu, l.............. Cu 15: a. Nitrogenase d b lm bt hot bi........, v vy cc t bo vi khun c kh nng c nh nitrogen hoc l phi h hp mnh vng b mt t bo gi cho phn bn trong t bo vng mt oxy (chng hn ...........................). b. Protein c chuyn ho thnh cc peptid bi cc enzym c tn l........, cn cc peptid c phn gii thnh cc aminoacid c tn l................ Cu 16: a. Qu trnh phn nitrat ho l mt qu trnh c hi i vi nng nghip, bi v......... b. S kh nitrogen dng kh thnh amoniac c tn l.................... c. Phc h enzym chu trch nhim i vi s c nh nitrogen cng sinh c tn l.... Cu 17: a. S phn nitrat ho xy ra mnh nhng ni oxy........................... b. Chi vi khun oxy ho amoniac thnh nitrite c tip u ng......, trong khi Chi vi khun oxy ho nitrite thnh nitrate c tip u ng............. c. Mt vi khun t sinh bo t sng k kh c nghin cu nhiu do kh nng c nh nitrogen mnh ca n c tn l.............................. Cu 18: a. Vi khun c nh nitrogen sng cng sinh trong bo hoa du thuc bn ....(1).... sinh oxy v c tip dinh dng l.................. b. Vi khun khng lu hunh mu ta hay gp trong cc ngun nc t nhin thuc bn............. sinh oxy v c tip dinh dng l................ c.Chi no trong cc chi sau y cha cc loi c nh nitrogen sng cng sinh: Rhizobium/ Anabaena/ Clostridium/ Frankia/ Azotobacter. Cu 19: Bt c lc no cng c th tm thy trn c th ca mt ngi trng thnh t nht............. t bo vi sinh vt, a s trong chng l..............v chng c gi l...................ca c th con ngi. Cu 20:
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Mt s vi sinh vt sng trn da, song a s sng trn cc b mt........ca c th, l.............................bao ph bn trong mi, ming, ng h hp trn, ng rut, v ng niu sinh dc. Cu 21: ................khng c coi l mt b phn ca khu h bnh thng. Cu 22: Nhiu c quan v v tr bn trong nh...................................................khng cha vi sinh vt. Cu 23: Nu hin tng ..............cn s c mt ca mm bnh sau s gii phng c t ca mnh vo cc t chc th hin tng.............ch l hu qu ca s tip xc gia c t vi t chc m khng nht thit cn s c mt ca mm bnh. Cu 24: Trng hp ng c thc n c tn l botulism (lit do ngoi c t tc ng ln h thn kinh) khng phi l mt s nhim trng m l mt s................... Cu 25: Mt bnh nhim khun c th c truyn bng ............, tc l s tip xc c hiu qu gia mt c th b nhim v mt c th khe mnh. Khng c nhm ln phng thc truyn bnh ny vi .......hoc ...... l cc phng thc truyn qua mt ng vt trung gian c tn l .............., gia mt c th b nhim khun vi mt c th khe mnh. Cu 26: Nu s...............gy nn mt s tham gia c hiu qu ca c th trong vic hnh thnh khng th, c gi l........................th s..............l mt phng php a vo c th cc khng th c tng hp t trc; l mt s min dch....................Trng hp u l trng hp tim chng .................., cn trng hp sau l trng hp tim chng ....................... Cu 27: Cht khng sinh l cc cht ha hc do VSV tit ra c kh nng......................... hoc ..............................ca cc VSV khc. Cu 28: Penicillin G ch yu hot ng chng li vi khun Gram..............., vi rng .............................Nhc im ch yu ca penicillin G l:................ Cu 29: a. Dng vaccine l a vo c th mt loi.......................ly t vi sinh vt gy bnh. b. Vaccin l vi sinh vt sng c ly t vi sinh vt sng mt...... c. Gii c t c sn xut t................... ca vi sinh vt. d. Mun phng dch c kt qu, vaccin phi c dng cho ....% i tng cm th. h.Vaccine c dng cho nhng ngi............. ca c th khe mnh
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Cu 30: Tr nh v ngi cao tui d mn cm vi cc bnh nhim trng v h hoc l h thng min dch........................hoc l........................ Cu 31: a. Mc ch ca kh trng Tyndal l tiu dit cc vi khun mang................... nhit ...........Nguyn l ca phng php l cho cc ...........ny................thnh...............ri sau un si git cht cc th .................. b.Trong k thut xc nh s lng t bo bng phng php nui trn mi trng rn, ngi ta gi khun lc l........(l ch vit tt ca.....) v rng khng phi bao gi n cng c mc ln t mt t bo....... 2. Hy tr li bng ng () hoc sai (S) Cu 32: Trong nhng s khng nh sau y, u l s khng nh ng: a.Nng lng gii phng trong d ho c s dng trc tip cho s tng hp cc thnh phn t bo. b. ng ho cung cp cc vin gch xy dng tng hp cc thnh phn t bo. c. S tng hp cc thnh phn t bo l mt qu trnh gii phng nng lng. Cu 33: Mt cht khng sinh c hot ph hp l cht khng sinh: a. Ch c tc dng ln cc vi khun Gram m b. Ch c tc dng ln cc vi khun Gram dng c. Ch c tc dng ln cc vi khun Gram dng ln vi khun Gram m d. Ch tc ng ln mt nhm ring bit, thm ch ln mt hoc vi loi vi khun. e. c s dng rng ri hn trong thc tin y hc so vi cc cht khng sinh c hot ph rng. Cu 34: a. Vi khun l nhng t bo b. Tt c cu khun u di ng c. Mi vi khun gy bnh u c th i hnh d. Sau khi b thoi hnh, vi khun c th tr v hnh th bnh thng nu iu kin thch hp. Cu 35: Nhng nhn nh no di y l ng vi t bo vi khun a. Nhn c phn cch vi phn cn li bi mng nhn b. Vt cht di truyn l DNA kt hp vi protein histon c. Khng c mng nhn d. Vt cht di truyn l DNA khng kt hp vi protein histon
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Cu 36: a. Acid teichoic l thnh phn c trng ca t bo vi khun Gram dng. b. S d c tn gi "sao chp bn bo th" v phng thc ny cho php bo ton nguyn vn 50% lng DNA c mt trong t bo m. c. Tc nhn gy bnh giang mai Treponema pallidum chuyn ng rt nhanh nh h thng tin mao bao quanh c th gi l chu mao. d. c im c trng ca mt ni c t l n lin kt vi t bo vi khun v s khng khi no c t bo tit ra. Cu 37: a. H hp k kh l qu trnh oxy ha thu nhn nng lng trong cht nhn in t cui cng l oxy lin kt. b. Cht nhn in t cui cng trong ln men ethanol l acetaldehyte. c. cc vi khun sinh methan thnh t bo cha peptodoglycan. Cu 38: Tm s khng nh sai Phng trnh: Glucose + ADP + Pvc a. Saccharomyces cerevisiae b. Sarcina ventriculi c. Zymomonas mobylis d. Escherichia coli 3. Sp xp s m t bn phi sao cho ph hp vi cc thut ng bn tri Cu 39: Bin np Ti np Tip hp Hfr Alen Cu 40: c tnh (Virulence) Bnh (Disease) a.Kh nng ca mt VSV gy nn mt bnh bnh b.VSV, c th bt u mt bnh mt c th khe mnh a. Kiu chuyn gen trong mt t bo th nhn nhn cc gen t cc phn t DNA t do trong mi trng xung quanh. b. Kiu chuyn gen ph thuc vo s tip xc t bo - t bo. c. Cc gen tn ti cng mt v tr tng ng trn nhim sc th tng ng nhng khc v mt trng thi t bin. d. Kiu chuyn gen trong mt phage c dng lm phng tin mang DNA t vi khun ny sang vi khun khc. e. Mt t bo F( trong plasmid F c gn vo nhim sc th ca t bo vt ch. 2 Ethanol + 2CO2 + ATP l phng trnh biu din tng qut qu trnh ln men ru :
Kh nng gy bnh c.Mc v kh nng gy ra mt bnh mt tc nhn (Patogenicity) gy bnh
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Tc nhn gy bnh (Causative agent)
d. S ph hng cc chc nng ca c th do nhim trng
II. TR LI
1. a. cu to, dinh dng, sinh sn. b. Loi(species) Chi (genus) - H (family) - B (order) - Lp (class) - Ngnh (phylum) - Gii (kingdom) - Siu gii (superkingdom); tn kp, ting Latinh. 2. khng c cu trc t bo; DNA; RNA; cc t bo vt ch; sinh nng lng; sinh tng hp; genom. 3. li acid nucleic; DNA; RNA; v protein; capsid; lipid, hydratcarbon; protein. 4. a. DNA si kp; gn vo genom ca t bo ch b. gai; hydratcarbon; protein; th th (receptor). 5. a. t 13 n 19; b. mu c. vi khun; gy tan; con ci; tim tan. 6. a. Nhim sc th, Bacteriophage; b. viroid, da, khoai ty, c chua.. c.prion. 7. a. chuyn ha; gai nhn bit cc th th ca t bo m chng gy nhim. c. vim no cp tnh; t vong 8. c; f. 9. a. siu xon; gin xon; b. trn; spheroplas. 10. nhn thc; kitin; cellulose; lc lp; hoi sinh, k sinh, cng sinh; bng bo t; lng; roi. 11. a. Cht nhn; mng nhn; b. ti th, li ni cht, b my Golgi c. PG; peptidoglycan; d. 70S. 12. a. ngh; mt; b. khng nhit. 13. a. plasmid, b. khng vi cc cht khng sinh; sn xut cc cht c kh nng gy bnh; sn xut cc cht bacteriocin; to nn mt c tnh trao i cht vi khun. 14. a. amoniac; b. vi sinh vt; c. nitrogenase; d. hai; nhm cng sinh v nhm khng cng sinh. 15. a. oxy; Azotobacter;
Vi sinh vt
320
b. protease; peptidase. 16. a.lm mt nitrogen ca t v do vy, lm gim cht dinh dng cho sinh trng ca thc vt; b.s c nh nit phn t; c.nitrogenase. 17. a. vng mt; b. nitroso; nitro; c. Clostridium pasteurianum. 18. a. (c); quang dng v c; b.khng; quang dng hu c; c. Rhizobium, Anabaena, Frankia. 19. 100 nghn t; vi khun; khu h bnh thng. 20. trong; mng nhy. 21. cc virus k sinh ni bo. 22. dch no ty, mu, bng quang, t cung, vi Falop, tai gia, thn... 23. nhim c; ng c. 24. nhim c. 25. tip xc trc tip; cn, t; vector truyn bnh. 26. tim chng; min dch ch ng nhn to; tr liu huyt thanh hc; th ng nhn to; phng bnh; cha bnh. 27. c ch, tiu dit chn lc; km hm s nhn ln. 28. dng; Khng c tc dng vi vi khun Gram m. 29. a.khng nguyn; b. c tnh; c. ngoi c t; d. 80; h. kho mnh. 30. cha hon chnh; khng cn hiu qu na. 31. a. ni bo t; 1000C; bo t; ny mm; t bo mi; dinh dng. b. n v hnh thnh khun lc; CFU (colony forming unit); n c. 32. b. 33. d. 34. d. 35. c,d. 36. a; d. 37. a. ();b. (); c (S). 38. c. 39. a. bin np; b. tip hp; c. alen; d. ti np; e. Hfr. 40. a. kh nng gy bnh; b. tc nhn gy bnh; c. c tnh; d. bnh.
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TI LIU THAM KHO

I. TI LIU TING VIT
1. 2. 3. 4. 5. 6. 7. 8. 9. Kiu Hu nh. 1999, Gio trnh vi sinh vt hc cng nghip. NXB KHKT. H Ni. Kiu Hu nh, 2006, Gio trnh Vi sinh vt hc (l thuyt v bi tp gii sn) song ng Vit- Anh phn 1 v phn 2, NXB KH&KT H Ni, Nguyn Ln Dng, Nguyn nh Quyn, Pham Vn Ty, Vi sinh vt hc, 1997, NXBGD H Ni. Phm Thnh H. 2005, Nhp mn cng ngh sinh hc, NXBGD. Bin Vn Minh (ch bin), Kiu Hu nh, Phm Ngc Lan, Phm Hng Sn, Phm Vn Ty, Nguyn th Thu Thu, 2006, Vi sinh vt hc, NXB i hc Hu. L Thi Lin Thanh & L Vn Hong, 2002, Cng ngh ch bin sa v cc sn phm t sa. NXB Khoa hc v K thut H Ni. Nguyn nh Thng & Nguyn Thanh Hng, 2000, Cng ngh sn xut v kim tra cn etylic. NXB Khoa hc v K thut H Ni. Lng c Phm. 1998, Cng ngh vi sinh. NXB Nng nghip. H Ni L Xun Phng. 2001, Vi sinh vt cng nghip. NXB Xy dng. H Ni.
10. L Ngc T (Cb), Ha sinh Cng nghip. NXB Khoa hc v K thut H Ni, 2005. 11. Wolfgang Fritche. 1997, C s ho sinh ca vi sinh vt hc cng nghip (Kiu Hu nh v Ng T Thnh dch). NXB KHKT.
II. TI LIU TING NC NGOI

[1] Aiba S., Hemphrey A. E. and Millis F. F.. 1973, Biochemical Engineering. Second Edition. Academic Press. [2] Lansing M. Prescott, john P. Harley, onal A. Klein, 2005, Microbiology. Mc Graw Hill. [3] Schlegel, H,G., 1992, Allgemeine Mikrobiologie, 7. Auflage, Thieme Verlag Stuttgart New York.
III. CC THNG TN T MNG

[1] http://www.google.com.vn/; http://vietsciences.free.fr/design/home.htm ..... [2] http://vietsciences.free.fr v http://vietsciences.net Nguyn Ln Dng [3] http://vietsciences.net v http://vietsciences.free.fr V Th Diu Hng [4] http://vietsciences.free.fr Phm Vn Tun [5] http://www.moet.gov.vn.
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1

BIN VN MINH (CH BIN)

Nicotinamid adenine dinucleotide phosphat dng kh Pentose phosphate Xylulose-5-phosphate

Chng 1: Vai tr ca vi sinh vt trong sinh gii

VAI TR CA VI SINH VT TRONG SINH GII

1. Khi nim chung v vi sinh vt hc v vi sinh vt

2. Cc c im chung ca vi sinh vt.

Hnh 1.1: Cc phng php quan st th gii sng (t nguyn t n t bo)

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Vi khun Escherichia coli

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Vt tch vi khun lam Cyanobacteria cch y 3,5 t nm

Vt tch Gloeodiniopsis cch y 1,5 t nm

Vt tch Palaeolyngbya cch y 950 triu nm

Hnh 1.3. Cc vi sinh vt ho thch

3. Cc nhm i tng vi sinh vt hc

Chng 1: Vai tr ca vi sinh vt trong sinh gii

5. Vai tr ca vi sinh vt.

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Hnh 1.5. ng dng ca vi sinh vt trong cng nghip

II. S LC LCH S PHT TRIN CA VI SINH VT HC

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

III. CCH C (PHT M) CA CH LATINH

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Bng 1.1: Cch c (pht m) ca ch Latinh

injectio in-ch-xi- (thuc tim)

quinque quanh-qu (nm)

amylum a-my-lum (tinh bt) Wolfram vn-ph-ram wedelia u---li-a (cy si t)

Bng 1.2. Quy tc vn phm Latinh bin i t s t sang s nhiu

Chng 1: Vai tr ca vi sinh vt trong sinh gii

IV. H THNG SINH GII V V TR CA CC NHM VI SINH VT

Chng 1: Vai tr ca vi sinh vt trong sinh gii

2. Mt s h thng phn loi

Hnh 1.28. Phn loi th gii sng

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Hnh 1.30: H thng 3 lnh gii ca Carl R. Woese v cng s, 1990

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Siu gii Acaryote Prokaryote Eukaryote

Cc siu gii ny c sp xp theo 3 mc phc tp ca c th sng

Chng 1: Vai tr ca vi sinh vt trong sinh gii

3. Nhng sai khc gia cc t bo Prokaryote v Eukaryote

Chng 1: Vai tr ca vi sinh vt trong sinh gii

V. VAI TR CA VI SINH VT TRONG I SNG

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Chng 1: Vai tr ca vi sinh vt trong sinh gii

VI. CC PHNG PHP V K THUT NGHIN CU

2 3 Hnh 1.36 : Mt s loi knh hin vi

Chng 1: Vai tr ca vi sinh vt trong sinh gii

+Nhum c bit -Nhum m (negative)

Chng 1: Vai tr ca vi sinh vt trong sinh gii

Hnh 1.39. Cc bc tin hnh nhum Gram v v d minh ho kt qu.

Chng 1: Vai tr ca vi sinh vt trong sinh gii

2. Phng php tch ly cc thnh phn ca t bo

3.Phng php nghin cu tnh a dng ca VSV

Chng 1: Vai tr ca vi sinh vt trong sinh gii