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QU TRNH LN MEN???
1 Ln men l mt qu trnh sinh hc, m phn ng oxy ha kh xy ra trong t bo vsv, trong iu kin khng c cht oxy ha no. Mt phn cht hu c b phn gii thnh cht kh, phn cn li b phn gii thnh cht oxy ha 2 Nghin cu qu trnh ln men l NC c im sinh l-ha sinh v hot ng sng ca mt chng VSV 3
Ty thuc vo c im sinh l ca VSV i vi oxy, ta coi qu trnh l hiu kh hay k kh khng bt buc
Ln men hiu kh c thc hin nh hai phng php c bn: nui cy b mt v nui cy b su (nui cy chm)
u im
Cc thit b ln men chm d c kh ho, t ng ho
Nhc im
i hi trang b k thut cao, d b nhim trng ton b Trong ln men chm cn phi khuy v sc kh lin tc
LN MEN GIN ON
VSV sinh trng n chng no mt thnh phn ch yu ca mi trng dinh dng b gii hn
C IM
L qu trnh nui cy VSV m mi trng dinh dng ch c cho vo lc u v khng thm cht dinh dng cho n khi kt thc qu trnh nui cy
LN MEN GIN ON
GIAI ON TIM PHT
Trong qu trnh nui cy, VSV sinh trng v pht trin theo mt quy lut gm 4 giai on
GIAI ON LOGARIT
GIAI ON N NH
GIAI ON T VONG
CC YU T NH HNG
Lng ging cy: ni chung lng ging cy nhiu th pha lag ngn v ngc li. Trong cng nghip ln men t l cy ging thng mc 1/10
Thnh phn mi trng: mi trng c thnh phn dinh dng phong ph th pha lag ngn.
(a )- nh hng ca s hn ch cht dinh dng i vi sn lng chung ca vi sinh vt. Lc nng cao th sn lng chung s t ti n nh. (b)- nh hng ca s hn ch cht dinh dng ti tc sinh trng.
C IM
1
NGUYN NHN
Hn ch cht dinh dng. Vsv hiu kh thng b hn ch bi nng oxygen. Tch ly cc sn phm trao i cht c hi.
Nhng bin i
V nhng bin i khc Sinh ra protein i
S cht ca t bo c th nhanh hay chm, c lin quan n s t phn hay khng t phn
NGUYN NHN
iu kin bt li ca mi trng: nng cht dinh dng thp di mc cn thit cho hu qu lm gim trao i cht , phn hy dn dn cc cht d tr v cui cng dn n s cht hng lot ca t bo Qu trnh bo qun cc chng VSV
100 120
5 6
25 = 32 26 = 64
32 64
1,505 1,806
Logarit 2 v:
Tc sinh trng
Xc nh thi gian th h
S hin din ca VSV c th c nh lng bng nhiu phng php nh: m s lng t bo trc tip trn bung m hng cu bng knh hin vi quang hc hoc bng knh hin vi hunh quang.
M TRC TIP BNG BUNG Nhc im: M Khng xc nh c t bo cn sng hay HNG cht, d lm t bo VSV vi cc ht vt th CU khc c trong mi trng v dch huyn ph VSV phi c mt t bo phi ln.
m trc tip bng bung m hng cu Pha long mu cn m sao cho trong mi nh ca bung m c khong 5 - 10 t bo VSV. Sau nh 1 git mu ln bung knh v y li bng l knh, quan st di lnh hin vi.
N=
Trong :
.. (T bo/ml)
A - s t bo trung bnh trong 1 vung ln. 1000 - h s chuyn i t mm3 sang ml (1 ml = 1 cm3 = 103 mm3) F - h s pha long ca mu trc khi m V - Th tch ca 1 vung ln
Nhum VSV bng cc cht pht hunh quang nh: acridin cam (AODC), 4,6-dianidino-2-phenyl indol (DAPI), flourescent isothiocyanate (FITC). u im: Kt qu phn nh ng vi sinh khi.
My m khun lc t ng
Cc loi khun lc mc trn mng lc. Theo sch ca Prescott,Harley v Klein (2005) (a)- Tng s vi khun mc trn mi trng tiu chun, Dng ch th mu nhum khun lc cho d im; (b)- Dng mi trng thch hp kim tra nhm vi khun coliform c ngun gc t phn (khun lc bt mu xanh); (c)- Dng mi trng thch m-Endo xc nh vi khun E.coli v cc Coliform khc- khun lc c mu lc; (d)- Nm si v nm men mc trn mi trng Thch - Mch nha.
XC NH KHI LNG T BO
XC NH TRNG LNG KH CA T BO
ra t bo ri lm kh
Cn trng lng kh
O MT QUANG (OD)
O C
c ca dch t bo c th m bng quang ph k v c biu din bng n v hp th. S lng t bo lin quan mt thit vi c ca dch vi khun. T bo phi c khuy trn k trc khi a vo quang ph k o c ca dch treo t bo. y l pp rt thun li. trong thc ta thng o mt quang hc ca dch treo (dch huyn ph). o s lng vi sinh vt bng phng php o c.
O MT QUANG (OD)
O MT QUANG (OD)
Thng qua vic o hp th nh sng c th xc nh c sinh khi vi sinh vt
Khi
Lng t bo tng
Dn n
c tng
Mc tn x nh sng nhiu hn Tr s hp th nh sng tng
Lm cho
o c
O MT QUANG (OD)
Cng thc:
A = ln (I0 / I)
Vi: A : l mt quang ti bc sng . I0 : l cng sng trc khi qua mu. I : l cng sng sau khi qua mu.
XC NH NNG T BO
Cng thc:
C(mg/ml)= A* k * n
Vi: C: nng t bo n: h s pha long k: h s chuyn i bng 5x108