PHNG PHP XC NH TNG S VI SINH VT HIU

KH

- Tng s vi sinh vt hiu kh: l tng s tt c cc loi vi khun a kh ( iu kin

c oxy t do ) c trong mu thc phm.
- Ch s ny cho ta bit s b mc nhim khun ni chung ca cc mu thc
phm, khng c gi tr quyt nh kt lun mu t hay khng t tiu chun vi
sinh.
- Mun nh gi mu t hay khng t tiu chun th cn phi thm ch tiu vi
sinh ch danh v vi sinh gy bnh c trong mu.

I. NGUYN TC
S dng k thut a, m s khun lc mc trn mi trng thch dinh
dng ssau khi hiu kh nhit 30 37 oC trong thi gian t 24 48 gi. S
lng vi khun hiu kh trong 1ml mu thc phm kim nghim c tnh t s
khun lc m c trong cc hp nui cy theo cc nng pha long .
II. DNG C V MI TRNG
- a petri c ng knh t 90 100mm.
- ng nghim.
- Pipet 1ml, 10ml.
- Erlen 250ml, 100ml, cc thy tinh 100ml.
- Mi trng (Nutrien agar, plate count agar).
- Pepton m.
- Nc pepton.
III. CHUN B MI TRNG
- Dng c c ra sch, sy kh v tit trng vi ni hp Autoclave p sut
1,5at/ 1h.
- Chun b mi trng nui cy.
Mi trng thch thng glucoza :
Pepton 10g.
NaCl 5g.
Cao tht 5g.
Glucoza 1g.
Thch 15 20g.
 Nc ct 1000ml.
Mi trng thch pepton glucoza :
Trypton 5g.
Cao men 2,5 g.
Cao tht 5g.
Glucoza 1g.
Thch 15 20g.
Nc ct 1000ml.
- C th s dng cc mi trng c pha ch sn ca hng Sanofi.
- Cch pha ch : ha tan bt mi trng vo nc, un nh la, khuy u ha
tan cc cht n khi si. ngui 70 -800C cho mi trng vo ng nghim
c sy kh (khong 15ml). Hp tit khun 1at trong 20 pht.
ngui cho thch ng t nhin.
Bo qun lnh trong mi trng t lnh (5 10 0C).
- Tin hnh nui cy :
K thut ly mu :
Mu c ng nht v pha long.
Dng pipet 1ml v khun ht 1ml sa ti t mu c lc u, cho
vo ng nghim c cha sn 5 ml pepton m.
Pha long mu c nng nh cy sao cho sau khi nui cy trn
cc hp thch c s khun lc t 30 300 khun lc. Mi mu nui cy 2
3 nng lin tip (ty theo mc nhim ca mu, mi nng pha
long cy 2 a petri). Thi gian t khi pha long mu n khi thch
vo trn khng qu 20 pht.
Cy mu
Dng pipet 1ml v trng ht 1 ml mu cc nng pha long 1/
10; 1/ 100; 1/ 1000 cho vo gia a petri (ch m np h khng c m
ht).
thch dinh dng hp v trng ngui 45 -50 0C vo tng
a petri, nu dng thch bo qun lnh th phi un cch thy cho thch
tan ht, ngui 45 500C ri cho vo a petri c sn 1 ml mu.
 Trn u mi trng v mu bng cch xoay xui, xoay ngc nhiu
ln nh nhng (khng lm qu mnh thch b sng ra ngoi hoc dnh ln
np hp) ri cc a thch kh t nhin.
p ngc a petri thch li t m 30 370C (24 48 gi).
Sau 224 -48 gi, c kt qu : m s khun lc mc trn cc hp
thch cy bng nh sng t nhin hoc qua knh phng i c nh
sng n.
Chn cc hp c s khun lc t 15 300 khun lc m. 2 m
pha long lin tip, nu khun lc nhiu qu c th chia hp thnh 2
4 phn bng nhau m.
Kt qu:
Tnh s N khun lc vi khun hiu kh cho 1 ml sn phm bng
cch tnh cng thc trung bnh cng tng s khun lc ca cc
hp trn theo cng thc sau :

C
N =
(n1 + 0,1 n2). D

Trong : C l s khun lc m c trn cc a chn.

n1, n2 l s a 2 pha long lin tip chn
c.
d l h s pha long ca m pha long.
Lm trn kt qu c c, ch gi li 2 ch s c ngha v biu th
kt qu di dng thp phn gia 1,0 v 9,9 nhn vi 10n (n l s m).
 S M TNG S VI SINH VT HIU KH
1.Chun b ng nht mu:

9 ml mu sa + 10 ml nc pepton m

2. Pha long mu:

1ml 1ml 1ml

9ml nc pepton

1: 100 1: 1000 1: 100.000

3. Cy 1 ml vo a petri:

1ml 1ml 1ml

1ml 1ml 1ml

4. Nutrien Agar 15ml vo mi hp

5. 35oC trong 24 48 gi, lt p a li
6. m nhng a c s khun lc 30-300, ly trung bnh cng ca 2 a nhn vi nng
pha long.