XC NH TNG S NM MEN V NM MC TRONG THC PHM BNG PHNG PHP A (PDA) THEO TCVN 4884:2005 Nguyn tc: Tng

g s nm men v nm mc c m bng cch v trong iu kin hiu kh. I. Chun b: - Dng c: ng nghim, bao P.E v trng, kp, mung, n cn,micro pipet, que cy, bnh tam gic - Thit b: cn, my ng nht, my rung, t II. Ch : -Tt c nhng dng c tip xc vi mu, mi trng u phi c tit trng. - Thit b v b mt ni thc hin thao tc phi c kh trng bng n cn, gn gng, sch s thun tin cho vic thc hin thao tc. - Tt c cc thao tc phi c thc hin bn ngn la n cn. - Cc dng c cha mi trng v dch mu phi c gi kn, trnh tip xc vi khng kh qu lu. III. Cch tin hnh: Bc 1: ng nht mu * Chun b mu: + i vi mu rn: Cn 10g mu (tht) trong ti P.E v trng. + i vi mu lng: dng ng ht ht 10ml dch mu cho vo bnh tam gic v trng Sau cho 90ml nc mui sinh l v trng vo mu c cn, ong. * ng nht mu + mu lng: lc k. + Mu rn: ng nht bng my Stomacher 30. Ta c dch mu c nng 10-1.

Bc 2: Pha long mu - Dng micropipet v trng ht 1ml dch mu c nng 10 -1 cho vo ng nghim cha 9ml nc mui sinh l v trng c dch mu c nng 10-2. Lm tng t pha long vi cc nng tip theo theo dy thp phn 10-3, 10-4. - Pha long mu thnh cc nng khc nhau tu theo lng VSV c trong mu. * ch : - s dng 1 micropipette v trng/1 nng pha long. - Rung lc u dch mu trc khi pha long nng k tip bng my rung. Bc 3: Nui cy * C 2 cch nui cy: a v cy trang. - Phng php a: + Dng micropipet ht 1ml mu pha long cho vo a petri d v trng ( mi nng 2 a ), lm lin tip 3 nng . + Sau 10-15ml mi trng PDA nu vo mi a sao cho va ,lc xoay u d ngui , lt ngc a v em i . - Phng php cy trang: 10-15ml mi trng PDA nu vo 6 a petri v i mi trng ng li mi trang tri iu dch mu trn mi trng. + Dng micropipet v trng ht 0,1 ml cho vo a mi trng, sau xung b mt phng xoay a cng v ngc kim ng h, sau khi trang tri u dch mu, kh b mt mi trng 15-20 ri mi em i 37oC/24h. Ch : + phi lt ngc a trc khi em i nhit 40-45oC vn cn hin tng bc hi nc trn b mt a nu khng lt ngc a nu khng lt p a th hi nc s ngng t v rt trn b mt mi trng gy hin tng mc long.

+ mi trng va , khng qu nhiu gy tn km, khng qu mng gy nn hin tng mc chng cho ln nhau. Bc 4: m khun lc: - im tt c khun lc mc trn mi trng nu nhiu th m bng my m khun lc -Khun lc mc loang tnh l mt khun lc -Chn a c s khun lc l 25-250 hoc 30-300 Bc 5: Tnh kt qu: Tng s nm men v nm mc: A= (CFU/ml) Vi: A: tng s nm men v nm mc m c 2 nng lin tip n: s a ti mi nng V:th tch dch mu nui cy vo a d:nng d pha long Kt qu: kt qu Nng 10-8 10-9 a 1 848 752 a 2 876 804 Tng s nm men, mc : A= =1,49.1011 (cfu/ml) KT LUN: Mu tht trn khng t tiu chun vi sinh trong thc phm (tng s nm men v nm mc: 1,49.1011cfu/ml) XC NH TNG S VI SINH VT HIU KH BNG PETRIFILM I)Dng c: -ng ngim -tm petrifilm -micropipet 1ml v 10ml -n cn II)Ha cht: -mu sa chua -nc mui sinh l III)Tin hnh:

-Dng micropipet ht 1ml dung dch mu cn phn tch vo ng nghim c cha 9ml nc mui sinh l trong iu kin v trng. -Lc u ng nghim trong 1 pht -t tm petrifilm ln mt bn phng -Dng kp inox lt mng film nilon ph b mt a petrifilm -Dng micropipep ht 1ml dung dch mu v nh ln gia mng mi trng trong tm petrifilm -Lt cun mng nilon cn thn tr li trnh bt kh -Dng tm nha chn c mt phng bn di dn mng dung dch u khp mng mi trng trong vng cy. Dng mt lc n nh ln tm nha tri u trn vng cy v ly tm nha chn ra khi -Cn thn em petrifilm 35OC trong 24h -m s khun lc v tnh kt qu KT QU: S khun lc cc nng lin tip: nng 10-9 :48 10-10:52 KT LUN: mu sa chua khng t tiu chun theo quy nh