Professional Documents
Culture Documents
BI BO CO
AN GIANG, 2013
Mc lc
Ni dung
Trang
Mc lc ................................................................................................................ i
Danh sch bng .................................................................................................. ii
Danh sch hnh .................................................................................................. iii
Chng 1 t vn .......................................................................................... 1
Chng 2 Ni dung ............................................................................................ 2
2.1. nh ngha .................................................................................................................... 2
2.2. nh lng Coliforms, Coliforms chu nhit, Coliforms phn bng phng
php MPN ............................................................................................................................. 3
2.2.1. Khi nim ................................................................................................. 3
2.2.2. Nguyn tc ............................................................................................... 3
2.2.3. Mi trng v ha cht ............................................................................ 4
2.2.4. Quy trnh phn tch................................................................................... 4
2.2.5. Cch c kt qu ...................................................................................... 5
2.3. nh lng Coliforms, Coliforms phn bng phng php m khun lc 7
2.3.1. Khi nim ................................................................................................. 7
2.3.2. Nguyn tc ............................................................................................... 7
2.3.3. Mi trng v ha cht ............................................................................ 7
2.3.4. Quy trnh phn tch................................................................................... 7
2.3.5. Cch tnh kt qu ...................................................................................... 9
Chng 3 Kt lun v kin ngh ..................................................................... 11
3.1. Kt lun ....................................................................................................................... 11
3.2. Kin ngh..................................................................................................................... 11
Ti liu tham kho ............................................................................................ 12
Ph chng....................................................................................................... 13
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Chng 1
t vn
Ngy nay, khi cuc sng con ngi ngy cng c nng cao th iu m h
quan tm l v sinh n ung. Hng nm c hng trm ca ng c thc phm
phi nhp vin, gy hu qu nghim trng n sc khe cng ng. iu ny
khng ch ph bin Vit Nam m ngay c cc nc khc trn th gii cng
vy. Mt trong nhng nguyn nhn gy ra tnh trng ny l s hin din qu
mc cho php cc vi sinh vt gy hi trong thc phm.
Kim sot cht lng v sinh v an ton thc phm lun l mi quan tm ca
mi x hi, mi thi i, c bit l hin nay, khi cuc sng ca con ngi
ngy cng cao, cht lng mi trng sinh quyn ngy cng thp, ngha l
him ha t cc tc nhn l, ha v nht l sinh hc t mi trng vo thc
phm ang tr nn ngy mt ln. Trong xu th hi nhp hin nay, cc x
nghip ch bin thc phm nu mun sn phm ca mnh sc cnh tranh
khng ch trn th trng ni a m c trn cc th trng nhp khu ln vi
yu cu v sinh ngy cng mt khc khe. Do vy buc cc nh sn xut phi
c nhng qui trnh kim tra v sinh tht khc khe t c cc tiu chun
quc t, mt trong nhng ch tiu m h quan tm n l s lng
Coliforms trong cc sn phm ca mnh.
Coliforms l sinh vt ph bin trong ng tiu ha ca con ngi v ng
vt. hu ht chng tn ti mt cch t nhin khng gy hi cho con ngi. Tuy
nhin, trong mt s trng hp, nht l khi sinh l c th thay i, stress th
mt s dng Coliforms mang gen gy c c th gy bnh trn ngi v mt
s loi ng vt. Coliforms l mt ch tiu thng dng c dng nh gi
mc an ton trong v sinh thc phm. S hin din mt s lng nht nh
Coliforms trong thc phm l nh gi thc phm khng an ton cho ngi
s dng.
Chng 2
Ni dung
2.1. nh ngha
Coliforms l nhng trc khun gram m, khng sinh bo t, hiu kh hoc k
kh ty , c kh nng ln men lactose sinh acid l sinh hi 370C trong 24
48 gi. Trong thc t phn tch, coliforms cn c nh ngha l cc vi khun
c kh nng ln men sinh hi trong khong 48 gi khi c 370C trong
mi trng canh Lauryl Sulphate v canh Brilliant Green Lactose Bile Salt.
Nhm Coliforms hin din rng ri trong t nhin, trong rut ngi, ng vt.
Coliforms c xem l nhm vi sinh vt ch th: s lng hin din ca chng
trong thc phm, nc, hay cc loi mu mi trng c dng ch th kh
nng hin din ca cc vi sinh vt gy bnh khc. Nhiu nghin cu cho thy
rng khi s Coliforms ca thc phm cao th kh nng hin din ca cc vi
sinh vt gy bnh khc cng cao. Tuy nhin, mi lin h gia cc vi sinh vt
gy bnh v vi sinh vt ch th ny vn cn nhiu tranh ci. Nhm Coliforms
gm 4 ging l: Escherichia vi mt loi duy nht l E. coli, Citrobacter,
Klebsiella v Enterobacter. Tnh cht sinh ha c trng ca nhm ny c
th hin qua cc th nghim Indol (I), Methyl Red (MR), Voges-Proskauer
(VP) v Citrate (iC) thng c gi tm tt chung l IMViC.
Bng 1: Biu hin sinh ha cc ging ca Coliforms
Voges
Phn ng
Indol
Methyl Red
Citrat
Proskauer
Escherichia
+(-)
+
Citrobacter
-(+)
+
+
Klebsiella
-(+)
+
+
Enterobacter
-(+)
+
+
Ghi ch: + phn ng dng tnh, - phn ng m tnh, +(-): a s phn ng
dng tnh v (+): a s phn ng m tnh.
Coliforms chu nhit l nhng Coliforms c kh nng ln men lactose sinh hi
trong khong 24 gi khi c 440C trong mi trng canh EC. Coliforms
phn (Faecal Coliforms hay E. coli gi nh) l Coliforms chu nhit c kh
nng sinh indole khi c khong 24 gi 44,50C trong canh Trypton.
Coliforms phn l mt thnh phn ca h vi sinh ng rut ngi v cc
ng vt mu nng khc v c s dng ch th mc v sinh trong qu
trnh ch bin, bo qun, vn chuyn thc phm, nc ung cng nh ch
th s nhim phn trong mu mi trng.
Coliforms pht trin tt trn nhiu loi mi trng, nhiu loi thc phm. C
nhng nghin cu cho thy chng c th pht trin nhit thp n -2oC v
cao n 50oC. trong thc phm chng pht trin yu v rt chm 5oC tuy
cng c ti liu ghi nhn s pht trin ca chng 3-6oC.
Ngng pH Coliforms c th pht trin l 4,4 9.
Chng pht trin tt trn mi trng thch thng, cho nhng khun lc thy
sau 12-16 gi 37oC, pht trin tt rt nhiu loi thc phm trong iu kin
thch hp.
Coliforms gm 2 nhm:
- Coliforms c ngun gc t phn pht trin nhanh, khong 16 gi, trong
mi trng dinh dng 44oC, khng mc 4oC trong 30 ngy. L loi vi
khun a nhit, nhit thch hp nht l 41oC.
- Coliforms khng c ngun gc t phn, chng c ngun gc thy sinh
hay t t, mc nhanh 4oC trong 3-4 ngy v 10oC trong 1 ngy. Khng mc
41oC, 44oC c ch hon ton s pht trin ca tt c cc Coliforms khng
c ngun gc t phn
2.2. nh lng Coliforms, Coliforms chu nhit, Coliforms phn bng
phng php MPN
2.2.1. Khi nim
MPN l phng php dng nh gi mt vi sinh vt theo s c xc sut
ln nht ca lng vi sinh vt c trong mt th tch mu, vi chnh xc
tng i cao. Phng php ny cn gi l phng php pha long ti hn
hay phng php chun , chng da trn k thut nui cy vi sinh vt cn
nh lng trong mt mi trng lng thch hp vi mt s ln lp li nht
nh. Pha long mt s ln mu c cha vi sinh vt, sau kim tra xem ti
pha long no cn pht hin s c mt ca vi sinh vt cn kim tra.
2.2.2. Nguyn tc
S lng Coliforms, Coliforms chu nhit, Coliforms phn trong mu nc,
cha mt thp ca nhm vi khun ny c th c xc nh bng phng
php MPN (Most Probable Number). Phng php ny da vo nguyn tc
mu c pha long thnh mt dy thp phn (hai nng k tip nhau khc
nhau 10 ln); 3 hoc 5 mu c pha long thp phn lin tip c trong
ng nghim cha mi trng thch hp c ng by kh Durham. Mi nng
pha long c t 3-5 ng lp li. Theo di s sinh hi v i mu nh
tnh s hin din trong tng ng th nghim, y l cc ng dng tnh. Ghi
nhn s ng nghim cho phn ng dng tnh mi nng pha long v da
vo bng MPN suy ra s lng nhm vi sinh vt tng ng hin din trong
1ml mu nc ban u.
2.2.3. Mi trng v ha cht
- Mi trng lng Lauryl Sulphate Broth LSB (canh Lauryl Sulphate)
- Mi trng lng Brilliant Green Lactose Bile Salt (canh BGBL)
Cc mi trng lng trn c chun b trong cc ng nghim cha ng
Durham p ngc. Sau khi kh trng, ch s dng cc ng nghim khng c
bt kh bn trong ng Durham.
- Canh Tryptone
- Mi trng rn Simmon Citrate Agar (thch Simmon Citrate) mi trng
c cha sodium citrate l ngun carbon duy nht. Khi vi khun s dng
sodium citrate s kim ha mi trng v mi trng thch s c mu xanh.
- Dung dch nc mui pepton SPW (Saline Pepton Water)
- Thuc th Kovac,s
- Canh MR- VP
- Thuc th Methyl Red
Thuc th -napthol
2.2.4. Quy trnh phn tch
Thng tin v sn phm
Nc ung ng chai AQUA MOON sn xut ti c s An Hnh (a ch:
22D2, H Huy Tp, khm ng Thnh 6, phng M Phc, Long Xuyn, An
Giang).
2.2.4.1. nh lng Coliforms
Chun b mu nc ng chai AQUA MOON, pha long mu, c mu c
pha long 10-1.
Tun t cy 1ml dung dch mu pha long 10-1 vo 3 ng nghim ging
nhau, mi ng cha 10ml canh LSB. Thc hin tng t vi dung dch mu
pha long 10-2 v 10-3 .y l trng hp xc nh MPN bng h 3 dy
nng v 3 ng nghim lp li. Nu nghi ng s lng Coliforms trong mu
qu cao, phi s dng cc mu c bc pha long cao hn. cc ng nghim
37oC trong 48 gi. Ghi nhn s ng c sinh hi. Dng que cy vng cy
chuyn dch mu t cc ng LSB (+) sang cc ng c cha canh BGBL v
37oC trong 48 gi. Ghi nhn s ng cho kt qu (+) (c sinh hi) ng vi mi
pha long.
Cy vo ng canh EC
Cy vo ng canh BGBL
44,5 0,2oC
24 gi
37oC 1oC
48 gi
Th nghim Indol
m s ng canh EC (+) v
indol (+), tra bng MPN
Coliforms
Coliforms phn
Cy vo ng canh BGBL,
37oC, 24 48 gi
Cy vo ng canh EC,
44oC, 24 48 gi
Mt Coliforms
Th nghim Indol
Mt Coliforms phn
Hnh 2: Quy trnh nh lng Coliforms, Coliforms phn
bng phng php m khun lc
2.3.5. Cch tnh kt qu
Da vo s khun lc m c v t l khng nh, tnh mt ca Coliforms
v Coliforms phn theo cng thc sau:
A (CFU/g hay CFU/ml) =
xR
N: tng s khun lc m c
ni: s i c s khun lc c chn ti mi pha long
V: dung tch mu (ml) cy vo mi a
fi: pha long c s khun lc c chn ti cc a m
R: t l khng nh
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Chng 3
Kt lun v kin ngh
3.1. Kt lun
Coliforms l sinh vt ph bin trong ng tiu ha ca con ngi v ng
vt. khi gp iu kin thun li chng s gy hi cho con ngi nht l khi
sinh l c th thay i, stress
Coliforms l mt ch tiu thng dng c dng nh gi mc an ton
trong v sinh thc phm. S hin din mt s lng nht nh Coliforms trong
thc phm l nh gi thc phm khng an ton cho ngi s dng.
Coliforms c xem l nhm vi sinh vt ch th. S lng hin din ca chng
trong thc phm, trong nc c dng ch th cho kh nng hin din ca
cc vi sinh vt gy bnh khc. S lng Coliforms cao th kh nng hin din
ca vi sinh vt gy bnh khc cao. So vi hu ht cc vi khun gy bnh
ng rut, Coliforms c kh nng sng lu hn. Nu trong mt th tch nc
nht nh (100 mL) khng pht hin thy vi khun ch th ng rut ny th
c th dng lm nc ung hoc nc sinh hot.
3.2. Kin ngh
Do Coliforms hin din khp mi ni nn phi kim tra trn nhiu sn phm
khc nhau.
S dng thm nhiu phng php khc kim tra nh lng Coliforms,
Coliforms phn, Coliforms chu nhit.
Nn n ung hp v sinh, chn thc phm an ton trnh cc bnh ly nhim
ng rut.
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Ph chng
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