Professional Documents
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BPhm
CNG THNG
Nhm 6
2005130382
2005130267
2005130370
2005130263
2005130387
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Nhm 6
Nhm 6
Mssv
2005130382
2005130207
2005130370
2005130263
2005130387
Cng vic
Quy trnh nh tnh Salmonella v hon
thin bi word
c im Salmonella
Tng hp v vit bo co
Cc phng php truyn thng
Cc phng php hin i
MC LC
DANH SCH NHM V BN PHN CNG VIC NHM 4........................1
MC LC.............................................................................................................1
DANH MC HNH NH.....................................................................................1
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Nhm 6
1.1 Mt s nh ngha.....................................................................................1
1.2 C ch qu trnh oxy ha..........................................................................1
1.3 Nguyn nhn qu trnh oxy ha................................................................1
1.3.1 Do ch n ung ca bn.......................................................................1
1.3.2 Do nhim mi trng............................................................................1
1.3.3 Do li sng...............................................................................................1
1.3.4 Do Bc x in t.....................................................................................1
1.3.5 Bnh mn tnh...........................................................................................1
1.4 Hu qu ca cc qu trnh oxy ha...........................................................1
CHNG 2 TC DNG CA THC PHM CHC NNG I VI
VN OXY HA.............................................................................................1
2.1 Tc dng chng lo ha v ko di tui th.............................................1
2.1.1 Khi nim.................................................................................................1
2.1.2 Biu hin ca lo ha...............................................................................1
2.1.3 Tc dng...................................................................................................1
2.2 Tc dng to sc khe sung mn..............................................................1
2.2.1 Khi nim.................................................................................................1
2.2.2 Cc yu t to nn sc khe sung mn.....................................................1
2.2.3 Tc dng...................................................................................................1
2.3 Tc dng tng sc khng, gim nguy c bnh tt................................1
2.3.1 Khi nim.................................................................................................1
2.3.2 Phn loi sc khng.............................................................................1
2.3.3 Tc dng lm tng sc khng, gim nguy c bnh tt........................1
2.4 Tc dng h tr lm p cho con ngi...................................................1
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Nhm 6
DANH MC HNH NH
Hnh 1.1. Cc tc nhn gy ra qu trnh oxy ha..................................................1
Hnh 3.1.Thc phm chc nng Enjuvenate Unicity............................................1
Hnh 3.2. Thc phm chc nng Fine Pure Collagen...........................................1
Hnh 3.3. Thc phm chc nng Ageloc Vitality Nuskin.....................................1
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Nhm 6
DANH MC VIT TT
TPCN: Thc phm chc nng
LI M U
Nhng nm gn y vn v sinh an ton thc phm lun l tm im ca cc
phng tin truyn thng i chng. Ngy cng c nhiu v ng c thc, nht l vn
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Nhm 6
S.pullorum nm 1909. Trc y ngi ta cho rng y l hai loi vi khun gy ra hai
bnh khc nhau ln gi chung l bnh ph thng hn g Typhus avium) v cn
bnh c tn chung l S.gallinarum - pullorum.
Nm l896 C.Archard v Rbensauded phn lp c vi khun Sparatyphi
equi v Sparatyphi bacilus. Ngy nay vi khun ny c gi tn l Sparatyphi B v
n nm 1898, Sparatyphi A tm c do N.Guyn v H Keyser.
Gii:
Bacteria
Ngnh: Proteobacteria
Lp: Gamma
B:
proteobacteria
Enterobacteriales
H:
Enterobacteriaceae
Hin nay theo phn
loi ca h thng Vin
Chi:
Salmonella
Pasteur th gii, Trung
tm Kim sot v ngn
nga dch bnh ca M Loi: Salmonella bongori
CDC (Control and Prevent
Salmonella
enterica
Disease Center), ging
Salmonella c chia lm
hai loi: S. enterica v S. bongori. Trong S. bongori gm tt c cc kiu huyt thanh
(serotype) cua loi ph s V, v S. enterica c chia lm 6 loi ph nh s: I, II. IIIa,
IIIb, IV v VI. Cc loi v loi ph ny c th phn bit c bng phan ng sinh ha.
Trong mi loi ph c nhiu kiu huyt thanh.
Cho n nay xc nh c trn 2463 kiu huyt thanh thuc ging Salmonella.
Theo h thng ca Kauffman White, cc kiu huyt thanh ny c chia da trn
khng nguyn thn O, khng nguyn tim mao H v khng nguyn b mt Vi. Phn
ln cc kiu huyt thanh c t tn theo cng thc khng nguyn, mt s khc c
t tn ring nh Enteritidis (S. enteritidis), Typhi (S. typhi), Paraphi ( S. paraphi),
Typhimurim ( S.typhimurium).
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Nhm 6
1.4 c im ca Salmonella
1.4.1 c im hnh thi
Salmonella l trc khun gram (-), hnh que, kch thc trung bnh t 2 3 x
0.5 1m, hiu kh v k kh ty nghi, c tin mao, c th di ng (tr S.gallinarum
v S.pullorum), khng to bo t, sinh acid t glucose v mantinol nhng khng len
mn saccharose, khng sinh indol v khng phn gii ur. Hu ht chng u dinh H 2S
(tr S.typhi), chng pht trin tt nhit 6C - 42C, thch hp nht 35C - 37C,
pH t 6 - 9 v thch hp nht pH = 7,2. nhit t 18C - 40C c th sng n
15 ngy.
Salmonella l vi khun k kh ty nghi, pht trin c trn cc mi trng
nui cy thng thng. Trn mi trng thch hp, vi khun s pht trin sau 24 gi.
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Khng nguyn
K
H
Tc ng
ca alcohol Formol 50%
50%
B ngn tr ngng
Khng
kt
Nhy cm
Khng
Khng
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Nhm 6
Kt qu
(+)
(+)
(+)
S dng Citrate
(+)
Sinh H2S
(+)
S dng Lactose
S dng Saccharose
H enzyme:
+ Catalase
+ Beta -galactosidase
+ Urease
+ Decarboxylase Lysine
(LDC)
Ornithine (ODC)
Arginine dihydrolase
(ADH)
Desaminase Phenylalanine
(-)
(-)
Ghi ch
Km theo s tng hp acid
Tr mt s chng thuc loi s.
ty phi v s. gallinarnm
Tr s. typhi v s. paratyphi A
Tr mt s chng thuc loi s.
paratyphi A v s. choleraesuis
Tr s. anizonae
(+)
(-)
(-)
Tr mt s loi ph
(+)
T r s. paratyphi A
(+/-)
(-)
(-)
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Nhm 6
(-)
(+)
(+)
(-)
(-)
Ghi ch: (+): dng tnh; (-): m tnh; (+/-): c th dng tnh hay m tnh ty
chng.
1.4.5 Yu t c lc
Vi khun Salmonella c th tit ra 2 loi c t: Ngoi c t v ni c t.
Ni c t ca Salmonella rt mnh gm 2 loi: Gy xung huyt v mn lot, c
t rut gy c thn kinh, hn m, co git.
Ngoi c t ch pht hin khi ly vi khun c c tnh cao cho vo ti colodion ri
t vo bng chut lang nui, sau 4 ngy ly ra, ri li cy truyn nh vy t 5
n 10 ln, sau cng em lc, nc lc c kh nng gy bnh cho ng vt th
nghim. Ngoi c t ch hnh thnh trong iu kin invivo v nui cy k kh.
Ngoi c t tc ng vo thn kinh v rut.
4
Ni c t - Endotoxin
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Nhm 6
h tiu ho, h thng min dch; vi cc biu hin bnh l: Tc mch mu, gim
trng lc c thiu oxy m bo, toan huyt, ri lon tiu ho, mt tnh thm n...
Ni c t tc ng trc tip ln h thng min dch ca c th vt ch, kch
thch hnh thnh khng th.
LPS tc ng ln cc t bo tiu cu, gy st ni c t, theo c ch :
Gii phng cc cht hot ng mnh nh: Histamin.
Ngng kt cc tiu cu ng mch.
ng vn, tc mch qun.
LPS tc ng ln qu trnh trao i gluxit: LPS lm tng cng hot lc ca cc
men phn gii glucoz, cc men phn gii glycogen, lm gim hot lc cc men tham
gia qu trnh tng hp glycogen
5
c t ng rut
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Nhm 6
c t t bo
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C ch gy bnh thng hn
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Nhng bin chng khc nh vim phi cp,, vim mng no, vim gan, vim ty
xng cng c th gp.
Nhng bnh nhn qua khi c khong 5 - 10% vn tip tc thi vi khun qua
phn trong qu trnh hi phc v 1- 4% tr thnh ngi mang vi khun lu di do vi
khun vn tn ti trong ti mt. Tnh trng ny c th ko di n nhiu nm v h tr
thnh ngun mang bnh rt nguy him.
8
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Nhm 6
CHNG 2
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Bc tng sinh:
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Cc mi trng tng sinh chn lc dng d pht hin Salmonella trong mu thc
phm l Rappaport Vassiliadis (RV), Selenite Cystein Broth,Tetrathionate Mueler
Kaumanm Broth (TT)...nhng nghin cu gn y cho thy mi trng RV c th thay th
cho cc mi trng khc. Hin nay, ngi ta khuyn khch l nn dng t nht 2 loi mi
trng tng sinh chn lc pht hin tt c Serotype Salmonella nu c hin din trong mu.
11
Bc phn lp
Nhm tch v nhn dng Salmonella khi cc qun th vi sinh vt khc trong
mu. Nhiu loi mi trng rn khc nhau c s dng phn lp Salmonella. Hin
nay, vcc tiu chun v sinh an ton thc phm khuyn khch s dng t nht 2 loi
mi trng phn lp khc nhau tng cng kh nng pht hin tt c cc dng
Salmonella, c bit l mi trng XLD c s dng nhiu nht.
12
Bc khng nh
Bo co kt qu
Qui trnh kim nghim ny ch cho php phn tch nh tnh Salmonella, gip kt
lun c hay khng c Salmonella hin din trong 25 g mu, khng cho php phn bit
c cc dng Salmonella hin din trong mu. khng nh c tn chng
Salmonella phn lp c cn phi tin hnh th nghim ngng kt vi cc loi khng
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Nhm 6
Nguyn tc.
Mi trng thuc th
Mi trng R-V R10 (RAPPAPORT-VASSILIADIS R10 BROTH)
B test nhanh 3MTM PetrifilmTM Salmonella:
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Nhm 6
Thit b ph tr
Thit b trn mu
T m
Thit b m khun lc
Ni hp tit trng
17
Quy trnh
a.
Cc bc thc hin
Bo co kt qu th nghim.
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Nhm 6
Nguyn tc.
y l phng php pht hin nhanh Salmonella trong thc phm, sa, trng,
thc n chn nui v mu mi trng trong vng 37 gi da vo s pht trin v hnh
thnh khun lc c trng trong mi trng chn lc ca Salmonella. Mu s c
tng sinh Salmonella trong mi trng Salmonella Enrichment, sau c cy
chuyn sang mi trng trng IRIS Salmomella Agar 371oC vi thi gian 243
h. Cc khun lc c mu hng sau chnh l khun lc Salmonella.
19
Mi trng thuc th
B kit th nhanh IRIS SalmonellaR:
Salmonella Enrichment
IRIS Salmonella Supplement
IRIS Salmomella Agar
20
Thit b ph tr
Thit b trn mu
T m
Thit b m khun lc
Ni hp tit trng
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Nhm 6
Quy trnh
a
Cc bc thc hin
Bo co kt qu th nghim.
Nguyn tc.
Mi trng thuc th
Nc dng pha long mu
B kit th E.coli - MCC test:
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Nhm 6
Quy trnh
a
Cc bc thc hin
Bo co kt qu th nghim.
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Nhm 6
Nguyn tc
Quy trnh
a
Bc 1:
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Nhm 6
Bc 3: Phn ng PCR
Sn phm PCR sau khi khuch i c phn tch bng in di trn gel 1.5%
agarose trong dung dch m TBE 1X v chp bng my chp hnh gel Biorad UV
2000. Thang chun 100bp ca cng ty Fermentas c s dng c lng kch
thc on sn phm PCR
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Nhm 6
CHNG 3
QUY TRNH NH TNH SALMONELLA TRONG
SA V CC SN PHM SA
3.1 Phm vi p dng
Quy trnh ny tham chiu theo TCVN 6402:2007 dng pht hin Salmonella
trong sa v cc sn phm t sa.
3.2 Nguyn tc
Phng php ny ch dng nh tnh v kt lun pht hin hay khng pht
hin Salmonella trong mu sa v cc sn phm sa ly.
Quy trnh pht hin Salmonella bt buc phi tri qua nm giai on v s dng
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Nhm 6
Mc ch
Tng sinh s b
Phn lp
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Phc hi
Th nghim sinh ha khng
nh Salmonella
Nhm 6
Dung dch A
Dung dch B
Dung dch C
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Nhm 6
Mi trng c bn
Trypton
Lactoza
Dinatri hydro phosphat ngm 12 phn t nc (Na2HPO4.12H2O)
Natri hydro selenit
Nc
j.
L-Xystin
Mi trng c bn
Cao tht b
Pepton
Cao men
Dinatri hydrophosphat (Na2HPO4)
Natr dihydrophosphat (NaH2PO4)
Thch
Nc
k.
Lactoza
Sucroza
phenol
Nc
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Nhm 6
Lc sng
Nc
31
Xylose Lysine Desoxycholate (XLD)
Bt cao nm men
Natri clorua (NaCI)
Xyloza
Lactoza
Sucroza
L-Lyzin hydroclorua
Natri thiosulfat
St (III) amoni xitrat
Phenol
Natri deoxycholat
Thch
Nc
32
Tryptone Soy Agar (TSA)
Trypticase peptone
Phytone peptone
NaCI
Agar
Nc ct
33
Lysine Decacboxylase broth (LDC)
L-Lystin monohydroclorua
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Nhm 6
Cao men
Glucoza
Bromocresol ta
Nc
34
Voges-Proskauer (VP)
Pepton
Glucoza
Dinatri hydro phosphat (K2HPO4)
Nc
35
Ure Broth
a
Mi trng c bn
Pepton
Glucoza
Natri clorua
Kali dihydrooctophosphat (KH2PO4)
phenol
Thch
Nc
m. Dung dch ur
Ur
Nc
36
Trypton Water
Tryptone hoc trypicase
Nc ct
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Nhm 6
37
Kligler Iron Agar (KIA) / TSI
Cao tht
Cao men
Pepton
Natri clorua
Lactoza
Sucroza
Glucoza
St (II) xitrat
Natri thiosunfat
phenol
Thch
Nc
Thuc th pht hin -galactosidaza
38
a
n.
Toluen
Dung dch m
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Nhm 6
(371)oC/(243)h
3.5 Cc bc
m tnh
3.5.1 Chun b
tin hnh
mu
chun
b dung dch ban u, cho 25
g mu th vo 225 ml mi
trng tin tng sinh, l t l ca mu
th vi mi trng tin tng
sinh c quy nh trong phng php
ny. Nu phn mu th khc vi 25 g, th s dng mt lng mi trng tin tng sinh
cn thit to ra pha long 1/10 (khi lng/th tch).
39
Sn phm sa kh
Chun b mt bnh c np y cha 225 ml mi trng tin tng sinh .
Lactoza
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Nhm 6
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Kh
Nhm 6
trong 18 gi n 24 gi.
Trang 40
Nhm 6
Thch TSI
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Nhm 6
Cy b mt nghing ca thch
Mu vng lactose v/hoc sucrose dng tnh (s dng lactose v/hoc sucrose).
Mu hoc khng i mu lactose v sucrose m tnh (khng s dng lactose
cng nh khng s dng sucrose)
Cc khun lc Salmonella in hnh th hin tnh kim (mu ) trn b mt
nghing ca thch v khi cy m su mang tnh acid (mu vng) c sinh kh (bt kh)
v vi khong 90% trng hp sinh hydrosunfua (thch b en).
Khi Salmonella c lactoza dng tnh c phn lp, mt nghing thch TSI s
c mu vng. Do , vic khng nh s b cc Salmonella khng ch da vo kt qu
th trn thch TSI.
Thch Ure
Nguyn tc: s dng pht hin kh nng phn ct ure thnh ammoniac do
hot tnh ca enzyme urease t vi sinh vt v kt qu l mi trng b kim ha do
NH3 c to ra.
Cch cy mu: cy cy ria trn b mt nghing ca thch. . 37 1oC trong
243h. Nu phn ng dng tnh, vic phn gii ur s gii phng amoniac, lm i
mu phenol sang mu hoa hng v sau di chuyn sang mu tm. Phn ng
thng xy ra sau 2 gi n 4 gi.
1
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Nhm 6
Lysine decarboxylase
Trang 43
Control
Escherichia coli ATCC 25922
Serratia marcescens ATCC 8100
Salmonella Typhimurium ATCC 14028
Nhm 6
37oC v trong 243h, kim tra ng thng xuyn. Mu vng cho thy phn
ng dng tnh. Phn ng thng xut hin sau 20 pht. Nu s dng a giy bn
sn, th theo cc ch dn ca nh sn xut.
Hnh 3.7 So snh gia Salmonella v citrobacter trong phn ng ONPG
50
Nguyn tc: pht hin kh nng oxy ha tryptophan thnh cc dng ca indol:
indol, skatol (methyl indol ) v indol acetate.
Cch cy mu: cy khun lc nghi ng vo ng cha 5ml mi trng tryptone
water. 37 1oC trong 243h. Sau khi , thm 1ml thuc th kovacs, nu phn
Trang 44
Nhm 6
Voges-Proskauer
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Nhm 6
3.6 Kt qu
Pht hin(hay khng pht hin) Salmonella trong 25g mu rn hoc 25ml mu
lng.
3.6.1 Gii thch t cc th nghim sinh ho
Bng 3.1 Gii thch cc kt qu
Php th khng nh
Phn ng m tnh
hoc dng tnh
100
91,9
TSI lactoza
99,2 a
TSI sucroza
99,5
91,6
Phn gii ur
100
L-Lyxin decacboxyl
94,6
Phn ng -galactoxidaza
98,5
100
Phn ng indol
98,9
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Nhm 6
Cc phn ng huyt
thanh
Phn ng in hnh
Khng
Phn ng in hnh
Khng
Tt c phn ng m
tnh
C th l
Salmonella
Phn ng in hnh
Khng th c
C th l
Salmonella
Phn ng khng in
hnh
Khng
C th l
Salmonella
Phn ng khng in
hnh
Khng
Tt c phn ng m
tnh
Khng c coi l
Salmonella
Trang 47
Gii thch
Nhm 6
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