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THC HNH VI SINH I CNG

BO CO THC HNH BI 14 NH LNG TNG VI KHUN HIU KH TRONG THC PHM BNG PHNG PHP M KHUN LC
I. NGUYN TC Tng s vi khun hiu kh l tng s nhng vi khun thuc nhm vi khun hiu kh tn ti trong mi trng. Tng s cc nhm vi khun ny trong thc phm c th xc nh bng phng php nui cy tri ln b mt thch hoc bng phng php to hp . Thng qua s lng khun lc m c trn cc a peptri cho php xc nh c lng vi sinh vt cn c kh nng sinh trng trong mi trng trong mu ban u. m kt qu chnh xc th s vi khun trong a phi trong gii hn 25-250 khun lc. Nu vt qu gii hn th phi tin hnh pha long v chn nhng pha long ln hn. Ch tiu tng s vi sinh vt hiu kh c dng nh gi cht lng ca mu v vi sinh vt, nguy c h hng, thi hn bo qun ca sn phm, mc v sinh trong qu tr n h c h b i n , b o q u n s n p h m . II. DNG C, MI TRNG, HA CHT Mi trng s dng: Plate count agar (PCA), pH = 7.0 0.2 Casein peptone: 5.0g Cao nm men: 2.5g Dextrose: 1.0g Agar: 15.0g Nc ct 1000ml

Mi trng c nu si cho tan agar, phn phi vo bnh tam gic, hp tit trng 121oC/20 pht, m (45 50oC) phn phi vo cc a petri. Nc mui sinh l 0.9%.
1 TRN THANH TNG MSSV: 10243221| GVHD: Thy TRNG V ANH DNG

THC HNH VI SINH I CNG

Nc ct v trng. Cn 96o. Nc ma. III. TIN HNH TH NGHIM 1. LY MU Quy trnh ly mu c nhng yu cu sau: Ly cc mu c tnh cht i din, lng mu va . Dng c ly mu, cha mu phi v trng. Mu ly xong, phi phn tch ngay, khng c qu 24h. Mu ly phi c nhn ghi k hiu ng thi phi ghi vo s nhng c im ca mu v ni thu mu.

2. PHA LONG MU

Hnh 14.1 Pha long mu

TRN THANH TNG MSSV: 10243221| GVHD: Thy TRNG V ANH DNG

THC HNH VI SINH I CNG

3. QUY TRNH
Mu

ng nht

Pha long

Hp

Hp tri

37oC (24 72h)

m khun lc ( 25 n 250 )

Tnh kt qu

TRN THANH TNG MSSV: 10243221| GVHD: Thy TRNG V ANH DNG

THC HNH VI SINH I CNG

Hnh 14.2 Quy trnh to hp 4. CCH M CNG THC TNH 4.1. Cch m

Hnh 14.3 Cc thit b m khun lc


4 TRN THANH TNG MSSV: 10243221| GVHD: Thy TRNG V ANH DNG

THC HNH VI SINH I CNG


Cch m th cng: Ly bt ch k hai ng vung gc y hp petri v nh du th t tng vng I, II, III, IV. m s lng khun lc tng vng. Nh nh du cc khun lc m. 4.2. Cng thc tnh: N(CFU / g hay CFU / ml) = Trong : N: s t bo ( n v hnh thnh khun lc ) vi khun trong 1 g hay 1 ml mu (CFU: colony forming units); C: tng s khun lc m c trn cc hp petri chn (c s khun lc nm trong khong t 25 250 khun lc/a); ni: s hp petri cy ti pha long th I; di: h s pha long tng ng; v: th tch dch mu (ml) cy vo trong mi a. IV. NHNG IM CN LU Khi tin hnh pha long mu, cc dung dch phi c ht ng th tch yu cu. i vi phng php to hp , mi trng nui cy sau khi hp tit trng phi c lm ngui n 15 50oC trc khi vo a petri cha dung dch vi sinh vt. Cc a petri nui cy vi sinh vt phi c lt p trc khi nui trnh cc khun lc b mt mc loang. Trong qu trnh m khun lc, khng c m hp petri. Que tri thy tinh dng tri dung dch vi sinh vt c kh trng bng cch nhng vo cn 96o ri t, khng c h trc tip trn ngn la n cn. V. BO CO THC TP Trnh by phng php nh lng tng vi khun hiu kh trong mu bng phng php m khun lc. CY THEO PHNG PHP TO HP : Pha long dch huyn ph cc nng khc nhau: 10-3, 10-4, 10-5 cy mu. Ghi vo np a petri c mi trng thch cc thng tin: nng pha long, ngy cy.
5 TRN THANH TNG MSSV: 10243221| GVHD: Thy TRNG V ANH DNG

C ( n1vd1 ... ni vd i )

THC HNH VI SINH I CNG


Dng pipette v trng ly 1 ml dch huyn ph cho vo mi a petri ( v trng). Cho khong 15ml mi trng PCA nhit khong 45oC vo hp a petri. Xoay chm cho hn hp trn u. yn cho ngui. Lt ngc cho vo t m. Nui cy 30oC trong 72h. Mi mu cy ba nng . Mi nng cy ba hp petri. Sau ly ra kim tra kt qu. PHNG PHP CY B MT Dch huyn ph c chun b tng t nh phng php to hp . Mi trng PCA sau khi c hp tit trng, c lm ngui n khong 50 60oC sau vo a petri v trng. a petri cha mi trng c lm kh b mt bng cch sy trong t sy nhit 35oC hoc chun b trc 2 ngy. Dng pipette v trng ht chnh xc 0.1ml hoc 0.3 ml nh ln b mt mi trng trong a petri. Tri u dung dch vi sinh vt l b mt mi trng bng que tri tam gic thy tinh. Cc a petri c nhit phng 15-20 pht cho kh b mt. Lt ngc cho vo t m. Nui cy 30oC trong 72h. Mi mu cy 3 nng . Mi nng cy ba hp petri. Sau ly ra kim tra kt qu. So snh u v nhc im ca phng php to hp v phng php cy tri b mt. Phng php cy tri: u im: ng u cao (khun lc mc u) nn d nhn dng cc khun lc c trng Nhc im: o Thi gian tri lu o Th tch mu cy nh o D nhim Phng php to hp : u im: th tch mu cy ln, mt vi sinh vt cao, thao tc nhanh, d dng. Nhc im: o D lm vi sinh vt cht nu mi trng cha h xung nhit 450C.
6 TRN THANH TNG MSSV: 10243221| GVHD: Thy TRNG V ANH DNG

THC HNH VI SINH I CNG


o Khng xc nh c hnh dng khun lc nht nh. o Kh m do khun lc chng lp.

TRN THANH TNG MSSV: 10243221| GVHD: Thy TRNG V ANH DNG

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