TCVN TIEU CHUAN QUOC GIA TCVN 8128:2015 ISO 11133:2014 VI SINH VAT TRONG THU'C PHAM, THUC AN CHAN NUOI VA NUOC - CHUAN BI, SAN XUAT, BAO QUAN VA THU HIEU NANG CUA MOI TRUONG NUO! CAY Microbiology of food, animal feed and water — Preparation, production, storage and performance testing of culture media HA NOI - 2015TCVN 8128:2015 Myc Ie Loi néi dau LO gidi thigu 1 Pham vi ap dung ..... 2 Tailiguvign dn... 3. Thuat ngt va dinh nghia... 3.1. Thwat nage chung va dinh nghia. 3.2 Thuét ngOr thir higu nang .. 3.3 Thuat ngér v8 méi trvéng nudi cdy 3.4 Thuat ngo vé vi sinh vat thir nghiem 4 Quan ly dim bao chat lueng.. 4.1 He théng tai figu 42 Baoquan.. . 4.3 Chudn bj mdi truéng trong phéng thir nghiém 4.4 Bao quan va han sir dyng mdi trvdng da chudn bi 4.5 Chudn bj 44 sir-dung. 4.8 U dm mai trong dac trong cdc dia Petr... 4.7 Thai bd m6i truéng..... 5. Visinh vat thir nghigm 48 thirhigu nang. 5.1 Yéu chu chun; 5.2. Chon vi sinh vat-thir nghigm. 5.3. Bao quan va duy trl céc vi sinh vat thir nghiém 54 Visinh vat 8 thir higu nang. 6 Kiém soat cht lvgng va thir higu nang cia méi trong nudi cdy. 6.1 Yéu cau chung.. 8.2. Kiém sodt chat lung ly - héa. 6.3 Kiém soat chdt lugng vi sinh vat........... ‘Yeu cau chung di voi phép thir hiu nang vi sinh vat. 85 Banh gia higu nang va din giai cc két qua... 8.6 Méi trang khang dinh va thude thir...TCVN 8128:2015 7 Phuong phdp thir higu nang ca méi trvdng nudi cdy Bac. 7.1 Yéu cau chun 7.2. Phuong phap thé dinh lwong. 7.3. Thirnghiém moi trréng nudi cy si dung cho loc mang... 7.4 Phuong php thir ginn tinh, 3 Phuong phdp the higu n&ng cua moi trvéng nudi ody léng.. BA Yeu cu chung wesc 2 3.2. Phuong phép dinh lung trong éng nghiém 48 thir higu nang cla moi trvéng tng sinh léng {phwong phép pha loging a8 phan biét) 8.3 Phuong phdp dinh tinh trong éng nghiém 48 thér hiéu nang ctla mdi trudng lng chon loc .... 8.4 Phuong php éinh tinh trong éng nghiém don (46 duc) 48 thir higu nang ctia méi trong l6ng....42 9 Phuong phap thi’ higu ning ctia chat pha loding va méi trong van chuyén..... 9.1. Yéu cu chung... 9.2. Phuong phép thir nghigm dich pha loang 9.3 Phuong php thé moi trvéng van chuyén 410 Hé théng tai ligu vé cdc két qua thir nghiém..... 40.1 Théng tin tir nha san vu. 10.2 Truy xudt ngudn QC... 0 Phu luc A (tham khdo) Tan goi ed céc thanh phan méi tréng nudi cdy si dung trong phan tich vi sinh vat thye phdm, thire an chan nudi va nude, Phu luc B (quy inh) Chuan bj gée 461 chieng va giéng lam vie. Phu luc C (quy dinh) So dd cia cdc phurong phdp thi higu nang Phu luc D (tham kno) Vi dy va phigu ghi cdc két qua thir nghiém méi trréng nubi ody .. Phy lye E (quy dinh) Cac vi sinh vat thir nghiém va cdc tiéu chi higu nang d6i voi mél tréng nudi cdy khuan lac duge str dung cho vi sinh vat trong thyc pham. 55 Phu luc F (quy dinh) Cac vi sinh vat thi? nghigm va cdc ti6u chi hidu nang d6i vei moi truréng nudi cay khudn lac duge sir dung cho vi sinh vat trong nvéc..... 78, Phu luc G (quy dinh) St dung c4c biéu dé kiém chirng dé kiém soat phép thir dinh long méi trrding nudi ody dic... 87 Phu iyc H (tham khdo) Bam bdo chit long otla mdi trudng nudi cy — Xt ly sy 6. 98 97 Sy: tye | (tham khdo) Phép thir dinh lugng mal trudng nudi cdy long. a sue Jey dy Re dh ce php nid ning sn vt vn erg moby hn héa...101 ‘Thu myc tai fiéu tham Khdo ... 105TCVN 8128:2015 Léi néi dau TCVN 8128:2015 thay thd TCVN 8128-1:2009 va TCVN 8128-2:2009; TCVN 8128:2015 hoan toan tuong duong voi ISO 11133:2014; TOWN 8128:2015 do Ban ky thuat tidu chudn quéc gia TCVN/TCIF13 Phuong phdp phan tich va ly mau bién soan, Téng cuc Tiéu chudn Bo luéng Chat rong thdm dinh, B6 Khoa hoc va Céng nghé céng bd.TCVN 8128:2015 Ldi gidi thigu Tai tt ca cdc phong thir nghiém thyc hign kiém tra vi sinh, c&c myc tiéu chinh la 48 duy tri, hdi sinh, ‘phat trign, phat hién va/ho’c dinh krgng cdc gidng vi sinh vat. Mai tru’ng nudi cdy du¢c sik dung trong tt 4 cdc ky thuat nudi cy vi sinh vat truyan théng va cling ding cho nhidu ky thuat thay thé khac, Nhidu céng thire mdi truéng nudi cdy 66 ban sn trén thi tredng va durge dling cho cac myc dich tang trudng dc bigt, duce mo ta trong cae tai figu. Nhidu phép thir va nhidu quy trinh phy thuge vao méi truéng nuéi cdy cd thé cho cac két qua hyp Ij va tai lap. Cc yéu cdu v8 méi trong nudi cy cé thé dac this adi véi mau va cac sinh vat c&n phat hién. Do G6, mai trréng nudi cy 4p ung cdc tigu chi quy dinh 1a didu kién tién quyét 461 vai moi céng vige v8 vi sinh. Thir nghiém thich dang phai dugc thyc hign dé chizng minh: a) sy ch4p nhan cila tieng mé méi trvdng: b) mi trudng la “phi hop cho muc dich”, va c)_ méi trréng 6 thé cho cac két qua phis hop. Ba tiéu chi tran la mét phan thiét yéu cla cde quy trinh kiém soat chdt Iugng ndi bé, voi cdc tai Higu thich hp, sé cho phép gidm sat hiéu qué méi truréng nudi cdy va gép phan vao viée cung cp di liéu chinh xdc va @éng tin céy. Béi véi phép phan tich vi sinh vat dng tin cy thi didu eo ban 18 str dung méi trvéng nubi cy ¢6 chat wong 43 duge ching minh. D6i vai tat cd cdc mdi trréng quy dinh trong cae phwong phap chuan, c&n xac dinh cdc tigu chi chp nhan yéu odu téi thiéu 48 dam bao dd tin cay cia m@i trzéng. Khuyén céo rng trong phép xéc éinh céc dc tinh higu sudt clia mai truréng nudi cy cn thyc hign cdc phép thir theo tiéu chuan nay. Vic thiét lap cdc tigu chi higu sudt t6i thidu duge chp nhan rng rai 41 voi méi truéng nudi céy cd thé dln dén-san phdm cé cht lugng phi hgp hon va do 66 lam gidm mire 6 thir nghiém can thiét trong phang thir nghigm. Ngoai ra, céc tiéu chi ch4p nhan bang phurong phap quy dinh trong tidu chudn nay 06 thé duge tat ca cdc phing thir nghiém vi sinh str dung 4@ dénh gia tinh higu qua, chon Ige va/ho&e dae tinh chon loc lia méi trang nudi cy. Phan tich vi sinh vat trong thyc pham, thire an chan nudi va nude, cdc yéu cau cua tiéu chudn nay phai dugc wu tién trong vide danh gia chat lugng ota méi trvéng nudi ody,TIEU CHUAN QUOC GIA TCVN 8128:2015 Vi sinh vat trong thyrc pham, thie 4n chan nuéi va nuée - Chuan bi, san xuat, bao quan va ther hiéu nang cua mdi trvéng nudi cd4y Microbiology of food, animal feeding stuffs and water — Preparation, production, storage and performance testing of culture media 4 Pham vi p dung Tiéu chudn nay dua ra cc dinh nghia lién quan dén viée dam bao chat lurgng cola méi trréng mudi cay va quy dinh cde yéu cau déi véi viée chuan bj cdc mdi truréng nudi cy ding 4é phan tich vi sinh vat trong thy phdm, thire &n chain nudi va cdc mu tir c&c mdi trvéng san xut thye phdm ho&c thie &n chan nudi cting nhu tt 63 cdc loai nuréc sinh hoat ho&c durge sir dyng trong ché bién thyre phdm. Cac yéu cau nay 06 thé 4p dung dé chuin bi tat cd cac loai mdi trréng nudi cAy sir dyng trong cc phéng thir nghigm thyc hign phan tich vi sinh vat Tiéu chudn nay cling dua ra cdc tiéu chi va quy dinh cdc phwong php dé thir higu ndng cla méi trang nudi cy. Tiéu chudn nay 4p dung cho cdc nha san xudt nhu: — Cée td chee thuong mai san xudt va/hodc phan phéi méi trxéng pha ché s&n dé sir dung, méi trxéng hoan chinh khé hodc méi truréng bén hean chinh; — Céc té chive phi thong mai cung c4p mdi trréng cho cdc bén ther ba; = Céc phéng thi nghiém vi sinh tw chudn bi m6i trong nudi cdy 48 str dung. 2. Tal ligu vign dain Céc tai ligu vién dln sau rt cAn thiét cho viée ép dung tigu chudn nay. 86i véi cdc tal ligu vign din ghi nm céng bé thi 4p dung phién ban durgc néu. Béi voi c&c tai ligu vign dln khéng ghi nam céng b6 thi 4p dung phign ban méi nhdt, bao gdm cd cac sira déi, bd sung (néu c6).TCVN 8428:2015 TCVN 6404 (ISO 7218), Vi sinh vat trong the phém va thie &n chan nudi- Yéu cdu chung va hudng in kiém tra vi sinh vat. TCVN 6507-1 (ISO 6887-1), Vi sinh vat trong thye phdm va thie dn chan nudi— Chun bj méu tho, huyén pho ban dau vé c&e dung dich pha loaing thép phan dé kiém tra vi sinh vat — Phan 1: Cac nguyén tc chung dé chudn bj huyén phd ban dau va céc dung dich pha loaing thap phan. TCVN 6507-2 (ISO 6887-2), Vi sinh vat trong the phdm va thc Bn cin nudi — Chudn bj mau thy, huyén pha ban dau va cdc dung dich pha loding thép phén d8 kiém tra vi sinh vat — Phan 2: Cac nguyén tc oy thé d8 chudn bj cdc mu thit va sén phém thit. TCVN 6507-3 (ISO 6887-3), Vi sinh vat trong thye phém va thie dn chan nudi ~ Chuan bj mau thy, huyén phd ban dau va cde dung dich pha fo8ng thép phan o& icém za vi sini vat— Phan 3: Cac nguyén téc cu thé d8 chudn bi cdc mu thuy sén va san phém thuy sén_ TCVN 6507-4 (ISO 6887-4), Vi sinh vat trong thye phdm v2 chic an chan nudi—Chudn bj mdu thi, ‘uyén pha ban dau vé cdc dung dich pha long thép phan o8 xiém tre vi sinh vat — Prin 4: Cc nguyén téc cu thé a8 chudn bf céc san phém khéc v6i sifa va sn pham sira, mit va sin phdm thit thuy sn va san phdm thuy sén. TCVN 6507-5 (ISO 6887-5), Vi sinh vat trong thyre phdm va thie dn chin nubi - Chudn bj mau thi, huyén phu ban dau va dung dich pha lo8ng thap phn dé kiém tra vi sinh v&t — Phan 5: Céc nguyén tc ou thé 08 chudn bi méu sita va san phdm sita. TCVN 6507-6 (ISO 6887-8), Vi sinh vat trong thre phdm va thire &n chin nubi— Chun bj mu thi, huyén pho ban dau va dich pha long thép phan dé kiém ira vi sinh vat— Phan 6: C4c nguyén tac cy thé d& chudn bj mau duoc Hy ti giai doan s4n xudt ban dau, TCVN 9716 (ISO 8199), Chét luong nue - Hutng din chung vé.dém vi sinh vat bang nudi cay. ISO 7704, Water quality — Evaluation of membrane filters used for microbiological analyses (Chat lugng nuée - Dénh gié b6 loc mang duge ste dung trong phan tich vi sinh). 3 Thuat ng@ va dink nghia Trong tiéu chudn nay sty dung céc thuat ngd va dinh nghfa sau day: CHU THICH 1. ©Ru nay cung ep cd dinh nghia chung ldn quan adn dim bio chBt krgng cia mdi sr=ng AUDI chy WA cuNg ‘chp cf thudtngiian quan @&n thye hin thir nghigm, mol trvtmg qubi cy v8 vi sinh vat nghigen, CHU THICH2 Cac Bang E.2 va F.2 diva ra cfc gi thich v8 c&c thuat ngi viet tt tén gol més turemg.TCVN 8128:2015 3.1. Thu§t ngir chung va djnh nghia 344 Kiém soat chat luryng (quality contra!) Mét phan cia quan ly chat Iurgng tép trung vao thy hién céc yau cau chat lvgng. (CHU THICH 1: Xom Tai ig them kh (1 342 Mé méi trv@ng nudi cy (batch of culture medium) L6 méi truéng nudi cdy (lot of culture medium) ‘Don vj mdi truéng déng nhét va hoan toan cé thé truy xudt lién quan dén mét lugng xac dinh cla nguyén ligu, ban thanh ph4m hode thanh phém, déng nhdt vé kigu logi va chat long va duge san xudt trong mét giai doan xc dinh, ¢6 cling 86 ma (hodc sé 16) san xuét. 343 Co chét tao mau (chromogenic substrate) Co chat phat huynh quang (fluorogenic substrate) Co chat 6 chiva m6t nhém ch4t mauwinhém chat huynh quang va co chat dé nudi cy duge bei vi khudn hoge ndm, (CHU THICH 1: Sau khi tBch cor chét tao mauca chat phat huynh quang, chét maufhuynh quang durge gidl phéng va sn phim ‘cub cing cb maufhuih quang 66 th nhin thy duget6é thé duige phat hign bang cach sir dung Gn cir tim (UV). 3.2 Thugt ng@ thir higu nang 324 Higu nang méi trong nudi cdy (performance of culture media) Sy ap ting cia moi trréng nudi cy di v6 cdc vi sinh vat thir nghiém trong cdc didu kién xéc dinh. 322 Vi sinh vit dich (target microorganism) Visinh vat hoc nhém vi sinh vat can durgc phat hién hod dinh Iuvgng. 3.23 Vi sinh vat khéng phal dich (non-target microorganism) Visinh vat bi tee ché béi moi truréng va/hode diéu kién & hode khéng hign thi cde ac trung mong muén coba cdc vi sinh vat dich. 3.24 Higu sudt cda méi trvéng nudi edy (productivity of culture medium) Mée thu hdi vi sinh vat dich tir mdi trvéng nudi cdy trong céc didu kién xd¢ dinh.TCVN 8128:2015 3.2.6 Tinh chon loc cia méi trvgng nudi cdy (selectivity of culture medium) Mire 48 te ché cdc vi sinh vat khéng phai dich trén hodc trong mét mai trréng nuéi cy chon I9c trong cac didu kién quy dinh. 3.26 Tinh de thd cia méi treéng nudi cy (slectivity of culture medium) Tish ic hidu cila méi truréng nudi cdy (specificity of culture medium) rong cdc du kign xdc dinh, cdc vi sinh vat khéng phai dich khong hin thi cdc dc tinh tong tly nhur vi sinh vat dich, 3.3 Thuat ngir vé méi trréng nudi cdy 33t Moitnromg nusi cdy (culture medium) Su tao thanh ciia cdc chat & dang léng, ban d&c ho&c dang ao, c6 chiva cdc thanh phan ty nhién vavhodc ting hep dé gitip cho vige nhan lén (c6 hodc khéng c6 sy We ché mot sé vi sinh vat nhdt dinh), nhan biét hove duy tri sir s4ng cila cac vi sinh vat, CHO THICH Kai euge sir dung cling vbi ce ovm tir kha thi thust ng nay thurn go tk ‘mdi trrdng” (vi dy moi trvémg tng sim). 3.3.2. MOI trwéng nu6i cdy phan Jogi theo thanh phan 33.24 Moi tring xac dint v8 héa hoc (chemically defined medium) Mai trémg nuai cdy chi bao gbm céc thanh phan héa hoc xac dinh 4 biét vé cdu trac phan tiv va do tin Knit. 3.3.22 Méi treéng khéng xac dinh hode méi trur’ng khéng xéc dinh mot phan vé héa hoc (chemically undefined or partially undefined medium) Mai trrémg nudi cdy bao gdm toan bé hoe mét phan cilia nguyén ligu ty nhién, da ché bién ho&c cach hdc, c6 thanh phan héa hoc chura dug xéc dinh hoain toan CHO THICH 1: Cc ten goi 6b ve cde thanh phn khdng xc dinh v8 mat héa hoe duge sir dung trong mei trvémg musi chy ure guy ph rong Phy cA, 33.23 MGi trong nudi cdy sinh mau (chromogenic culture medium) Méi trueng nudi cdy phat huynh quang (fluorogenic culture medium) Méi treéng nudi c4y c6 chira mét hoa nhiéu co chat tao mau/co chat phat huynh quang. 10TCVN 8128:2015 CHU THICH 1: Mél tréng nubi cdy sinh mau tao thudn tign cho viéc nhgn biét cdc vi Khudn hoc ndm boi mau sée xéc Binh hol co dc idm hinh th (mot truémg nub cy phat ibn din inh). Mei tubng nub cy phat hyinh quang yeu edu quan sat ‘dudi d8n UV. Céc sin phim phén img sinh hba on thidt cho vigc anh gié moi tretmg nudl cy sinh mauiphat huynh quang, ‘thuréng 1a két qua hogt dng enzym cila ofc sinh vat nhl dinh, phy thugc rit nhidu vao vibe duy tri chinh xéo c&c didu kign ey thé (vl dy nur nhit 49, 69 pH, nding 6 oo cht, 3.3.3. Mdi truréng nudi c4y dugc phan logi theo trang thai vat ly 3.3.34 Mi trirémg léng (liquid medium) Méi truxéng nudi cdy bao gdm dung dich héa tan cia mét ho&c nhidu thanh phan, nhur nuréc pepton va canh thang dinh duémg, CHU THICH 1: Trang mt s6 még hop, cc hat rin uve cho vo ma trémg nubl ely dng, nhur rune ti. (CHU THICH2 : Mb trate dang Wing dung tong cfc dng, binh hog chai thurdng duge gail "canh thang”. 3.3.3.2 Mai trurémg dic (solid medium) Mi trrémg ban dc (semi-solid medium) Moi trréng King chira cdc ch&t lam déng d&c (vi dy: thach, gelatin) & cdc ndng a6 kteic nha. CHU THICH 1 : Do mol trxdmg agar-agar dong dic ouge sir dung rong r8i, nén thult not rot ngn “ager” Che) thrémg durge ‘str dung ding nghifa voi mdi trréng dc va lin quan dén “thach dém ota", CHU THICH 2 : Mol rubng dic cuge r6t vao céc dla Pet thrdmg durge gol “hach dm dia". Moi trutmg Bc dugc r6t vao cae ¢6ng hod chal nhd éhrge git theo vj trl nghiéng 48 cho mdi tring ding dc lsi Guge gol la "thech nghiéng” hoe "abe". Néu mdi trudng durge phan phbl vio day vat chira, tao thanh “hach dimg”. 3.3.4 Mél trurgng nudi cdy dye phan logl theo myc dich si? dung 3344 Mél-trvémg van chuyén (transport medium) Méitrréng dure thiét ké a8 bao thn va duy tri-kha nding sng ola vi sinh vat trong khiwan-gidnrrthige thoi gian tinh tir kie thu thap mu dén khi x fy mau trong phang thi nghiém. ‘VIDY: Ml truémg vén chuyén Stuart hode Amies. 3342 MOI truéng bio quan (preservation medium) Méitruéng due thiét ké 48 bao tn va duy tri sy tin tal cia céc vi sinh vat trong mét thei gian cai, nh bao vé c&c vi sinh vat chéng lai nhtmg anh hung bat I¢i 6 thé xay ra trong thé gian bao quan dai va cho phép phyc hdi sau giai dogn bao quan. ‘Vi DY: Mol tring tring Dorset, thach dinh duémg nghiéng. 1TCVN 8128:2015 3.3.4.3 MOi triréng pha loang (diluent medium) MGi truréng huydn phil (suspension medium) Méi truéng dug thiét ké d& tach cdc vi sinh vat tly sn phdm thir nghiém dang ac thanh dang lng vathode lam gidm néng 46 bing céch pha lofing ma khéng lam tang hoae fam dec ché vi sinh vat trong thoi gian tiép xuc. ‘VIDU: Dung dich mudi pepton. 3.3.44 Méi trong phye hdi (resuscitation medium) Moi truéng tao didu kign cho céc vi sinh vét bi te ché va bj hur hdng o6 thé phyc hdl va phat trién binh ‘thurdng ma khéng en thiét phai nhan lan. VIDU: Nude dem pepton . ‘CHU THICH 1: MBitrvéng phyc hi cing 06 né dure sk dung kam ma tréng tng sinh so BG, vi du: nade d8m pepton. 3.3.4.5 Mi trong tang sinh so’ bé (pre-enrichment medium) Méi trong ting sinh (enrichment medium) MGi trvéng dang lang, o6 thanh phan tao didu kign thugn Ioi cho vie nha [én cdc vi sinh vat. ‘VIDY: Canh thang trypton agu tong, 33.454 MGi trvéng ting sinh chon Igc (selective enrichment medium) Mi trurding tng sinh cho phép mot sé vi sinh vat nhdn Ién trong khi lai ec ché mot phan hog hoan toan sy ting truéng ciia cde vi sinh vat khée. VIDY: Mobi trong Rappaport: Vassitads pepton dau twong (RVS). 3.3.4.6.2 Méi tréng tang sinh khéng chon I9c (non-selective enrichment medium) Méi trréng tang sinh cho phép nhibu loi vi sinh vat phat trién. ‘VIDY: Canh thang tim no. 3.3.46 Mi truéng phan lp (isolation medium) Mai truréng daic hoac ban dac cho phép cde vi sinh vat phat trién. 33.464 Méi trixéng phan lap chon Igc (selective isolation medium) Méi truréng phan lap cho phép phat trién cc vi sinh vat dich cu thé, trong Khi lai tre ché hoan toan ho&e ‘mot phan céc vi sinh vat khac. ‘VIDY: Thach deoxycholat cefoperazon than edi bidn (Thach mCCD). 12TCVN 8128:2015 3.3.4.6.2 Mai trzémg phan lp khéng chon Igc (non-selective Isolation medium) 'MOi trdng phan lap khéng te ché chon loc cdc vi sinh vat. VIDU: Thach dnh dim. 3.3.46.3 Mdi trvémg nudi cdy chon Igc sinh mau (chromogenic selective culture medium) Ml tring nudi cdy chon Ioc phat huynh quang (fluorogenic selective culture medium) Méi trvéng nuéi cdy sinh mau/phat huynh quang ofing chtra cdc cht chon lec a8 tec ché hoan toan hose mét phan di kém véi hé vi sinh vat cd trong mau thi, do dé hé tre cho viéc phat hién chinh x4¢ cdc vi sinh vat dich. Vi DY: Thech TBX, mb trong MUGS. 3347 Mi trréng phan bigt (differenaal medium) Moi truréng dc trumg (characterization medium) Méi truéng cho phép thir nghigm mot hogc nhiéu dc tinh sinh Iy/sinh héa ca cdc vi sinh vat dé nhan biét ching, VIDY: Thach TRX, thech Lactose wii tepid 7a TTC. CHU THICH 1 : Ce méi zing phan bit 5 th8 die sir dung nhur mol trvémg phan Hep éurge gol l& mel frving phan lep/phsn Dit, v du nh thech deoxychoiat Wsin xylose (XLD),thach TTC lactose. 3.3.4.8 Méi trréng nhgn biét (identification medium) M@i trvéng dug thiét ké 68 tao ra phan tmg nhdn biét cy thé ma thuéng khéng yéu cau thém bat ky phép thir khang dinh nao. VIDU: Thach mgt azid aescutn, 3.3.49 Méi trong dinh hegng (enumeration medium) Méi trudng nudi cdy chon Ipc ho&e khong chon lec, cho phép dinh igng cc vi sinh vat. ViDY: Thach Baird Parker, chat chidt nim men. CHO THIGH 1: Mei trrémg cinh krong o6 thB bao gbm cic huge tinh cia mai tung tng sinh vB/mod phyc hl. 3.3.4.10 Méi truéng khang djnh (confirmation medium) Méi trvéng gép phan vao viéc nhn biét hodc xc dinh dc trung cilia vi sinh vat sau khi phuc hdi so bé vvasho§e ting sinh vafhodc phan tap. Vi DY: Thach sBt Kiger, 13TCVN 8128:2015 33.441 Moi trig c6 chira céc chit trung héa (medium containing neutralisers) Mai truémg van chuyén, méi truéng pha long ho&c méi trvréng nubi cdy cd chia cdc chit trung hoa 48 lam bat hoat chat tdy riva/ chat khir trung hodc cae chat diét sinh vat khdc. 33412 Méi trdng sir dyng cho nhiéu myc dich (medium having multiple uses) Méi tredng dure phan vao mét sé myc khac nhau. “VI DU: Thach mau la m6i wwéng phyc hdi theo 3.3.4.4, mdi tring phn Ip theo 3.4.4.6 va méi tratmg phan bigt theo 3.3.4.7 ir dung 68 phat hign tan mau. Nuée idm papon 18 chét pha long theo 3.8.4.3 vB mei tina tng sinh so bb theo 3.3.4.5. 3.3.4.13 Méi trréng adi cheng (reference medium) Mai trudng, thurémg 1a khong chon loc, 48 danh gia so sdnh va higu sudt dc lap cla méi zuéng can ther ghiém va digs ching minh fa phi hep cho viée st dung a8 kiém sot. ‘ViDU: Thach typton dau tweng (TSA). 3.3.5 Méi truréng nudi cdy duge phan toai theo phurong phép chuan bj 3.3.51 Méi trong dé sir dung ngay (ready-to-use medium) Mai trang Keng, @&c hoge ban dc due cung ofp trong cde dia, chai lo, éng nghiém hoe vat chira khdc, & dang 68 str dung ngay hodc 8 sir dung sau khi lam tan chay ho&e 68 sir dung ngay sau khi lam tan chay va bé sung. 3.3.6.14 Méi trong nudi cdy hoan chinh (finished culture medium) Méi trrdng & s&n sang dé nudi ody. 3.3.61.2 Moi trong sir dung ngay sau khi lam tan chay Iai (ready-to-use medium after remelting) Méi treving cn lam tan chay lai, vi du 8 si dung trong kf thuat d8 dia ho&c dé rét vao cdc dia Pati. 3.3.6.1.3 Méi trang sir dung ngay sau khi lam tan chay Iai va bé sung (ready-to-use medium after remetting ‘and supplementing) ‘Méi trrémg can lam tan chy lai, bd sung va phan phéi treéc khi str dung (truc 66 mdi truéng chua sin ‘sng dé sir dung). \ViDY: Thach sult xycioserin tryptose (TSC), thach Baird. Parker hois thach Frinagen huyét trong ths (RPP). 14TCVN 8128:2015 3.3.52 Mai trong diye chudn bj tie cac ché phdm khé ban sn (medium prepared from commercially dehydrated formulations) Mai tru’ng & dang khé yu cau bi nude va xtrIy true khi sir dung, & mét trong hai dang sau: — mdi trréing hoan chinh; = méi tréng khéng hoan chinh can bb sung truéc khi sir dung. VIDU: Dang bét, dang hat, sain phdim Ging khd. 3.3.6.3 MQi trong duge chudn bj tir cac thanh phan riéng 18 (medium prepared from individual component) Moi trrémg duge sn xudt béi mot phéng thir nghigm vi sinh hon toan tir cdc thanh phan riéng [6 34 Thuat ngi vd vi sinh vat thir nghigm 3.44 Sinh vat thir nghigm (test organism) Visinh vat thudng durgc st? dung 48 thir higu nang clia moi trvdng nudi cy. CHO THICH 1: Sinh vot thir nghigm durge xf dinh ip tho ngubn cba ching (xem 3.4.2 dén 3.4.7, 342 Chiing 441 cheng (reference strain) Visinh vat thu duc tryc tigp ttr mot bd suru tép chiing d6i chirng, tir bo suru t&p chiing oda Lién Goan thé giéi cdc Bao tang giéng vi sinh vat (WFC) hoc T8 chirc Bao tang giéng vi sinh vat chau Au (ECCO) va due xéc dinh it nh&t én mite chi va loai, durge phan logi va mé ta theo dc diém cilia ching va tét nhat 1a c6 ngudn géc tir thye phdm, thére &n ch&n nuéi ho&e mdi trvéng sn xudt thyre phaim hod thie an chain nudi hoa née. 3.4.3 Géc di ching (reference stock) Cac gidng riéng biét gidng hét nhau thu duge bang cdy chuy&n riéng ré tir ching d6i chéng trong phong thir nghigm hod c6 duge tir mét nha cung cp. 344 Giéng géc (stock culture) Cy chuyan tir gde adi chieng. 3.4.5 Gidéng fam vide (working culture) Cy chuyan tir abe d6i ching hode gidng géc hod chat abi ching, 48 durge chi¢ng nhan hoc khéng.TCVN 8128:2015 3.46 Ghat chudn (reference materia!) RM Vat lidu ¢6 chira mét long vi sinh vat 06 thd s6ng lai, dng nhat va én dinh aéi vai sé Ivgng vi sinh vat séng, 64 duge thiét lap 1a 48 phis hop véi mye dich st dung trong qué trinh dinh iugng. (CHU THICH 1 : Xem Tai ibu tham Kho [3 347 Chat chudn a diye chimg nhn (certified reference material) cRM Chat chudin duge dinh 13 dc dim bang quy trinn c6 higu Ive v8 do luring dé dinh long cac vi sinh vat s6ng, di kem gidy ching nan ghi ro sé trong quy dinh cia céc vi sinh vat s6ng, voi dO khong dam bao do va lién két chudn do hrémg. CHU THICH 1: Xem Tai ligu thar kngo TS 4 Quan ly dim bao chat hreng 4.1 H@ théng tai ligu 414 Tai ligu ti nha sin xudt hoge ngudi san xudt Nha sn xuat (cc t8 chirc thurong mai hoae phi thireng mai cung c&p méi trréng cho bén thir ba) cin 06 sn céc tai ligu sau day: —_ t8n méi truéng, cdc thanh phan riéng Id va cdc thanh phan bé sung bat ky, néu 66 va cdc ma sé s4n pham; bang dé ligu ky thuat, vi du: cong thire, myc dich sir dung, lung 48 day, néu 66; = dirligu an toan vaéJhoac méi nguy khi can, — sémé; pH dich oda m6i trudng hoan chinh; — théng tin bao quan va han str dung, — han sir dung dy dinh; ~ gidy ching nhan kiém soat chat hrgng, cdc wi sinh vat thr nghiém durge sir dung va cdc két qua phép thir higu nding theo céc tiéu chi chp nhan. 44.2 Chép nhgn giao san phim ‘B6i voi mBi m8 s4n pham (thanh phan hoae mdi trong nudi c4y), kim tra nhu sau: 18TCVN 8128:2015 nbn biét sn phdm; — tinh nguyén ven cla bao géi; = han dung cia san phim; = tai ligu dure cung cp. — 86 lugng don vj nhan duge Ghi lai ngay nhan san pham. 4.2 Bao quan 4.2.4 Yéu clu chung Trong mgi trvéng hyp, phai tuan thi cdc huéng dln cla nha san xudt, 4.2.2, Quan ly chdt lugng va kiém soat san phdm méi tréng khé va cdc thanh phan bé sung Mi truréng dang bot khd hoe dang hat dung trong vat chira cé np day kin. Cac chét bd sung chon gc khdc nhau ho&e cdc ch&t chdn doan dure cung cp 6 trang thai léng, bot ho&c déng khé. Vike dat mua méi truéng cn lap ké hoach quay vong dinh ky trong kho (nghTa la vao trréc-ra trudeo). Khi ma vat chira méi cn: ~ kim tra niém phong; — ghi ngay mé [an dau tién; - nhin dé danh gia lugng chira trong hop 44 mé. ‘Sau khi mé hép chtra méi, chét lurgng cia méi trréng cé thé phy thudc vao didu kién bdo quan. Chat legng méi trrdng khé bj gidm khi thy cé sy thay ddi dac tinh chay cla bét, tinh ddng nhét, von cyc, abi mau v.v... M6i trrémg khé a4 hit Am hodc thay thay @3i nhidu vé hinh thirc bén ngoai thi phai loai bé. Khi més chai dymng mdi truréing kh6, ghi lai ngay thang va néu ré thai gian bdo quan t6i da. 4.3 Chuan bj mdi treéng trong phéng thir nghigm 43.1 Yeu cdu chung Chuan bi chinh xéc méi trong nudi cdy ld mét trong c4c ure co ban 48 dam bao tinh toan dign trong kidm tra vi sinh vat va due chi ¥ aac biét. Chu y vé thyc hanh tét phdng thiy nghiém va céc huéng din cia nha san xuSt lin quan dén xir ty moi truéng khé va cdc thanh phan khdc, dae bigt cc thanh phAn ¢6 chiva cc vat liu déc hai, nghfa Ia muéi mat, natri azid, chat khang sinh ho&c cdc chat chon loc khdc.TCVN 8128:2015 Khi mdi :ruéng dugc chudn bj tir c&c ché phdm khd o6 ban sn, thi tudn thi chinh xe cdc huéng dan ca nha san xuat. Ghi lai t&t c& cc dé liéu lién quan, nhur khéi lurgng/thé tich, ma san phdm, sé higu !d hang, pH, naay chudn bi, digu kign khir tring, ngu6’ thye hign. ‘D4i voi méi trur’ng durge chuan bi tir cdc thanh phan riéng 18, thyc hign chinh xéc theo céng thie. Ghi lai t€t c& moi chi tiét va nhan biét day du (nghia la ma san phdm, sé hiéu 16 hang va han sir dung, néu c6 sn) clia tt cd cdc thanh phan duge st dung. Phu lye D néu vi dy vé théng tin nay. 4.3.2 Chat lugng cila cdc thanh phan méi trueng co ban Céng thire cia cae thanh phan méi trong co ban durge quy dinh trong cdc tiéu chudn cy thé (xem ‘Thu mye tai figu tham khdo). Khi s&n 6, néu r6 khél lweng phan tir va s6 CAS” cla héa chat trong cng thie. Trong mat s6 trréng hop, thanh phan cy thé (vi dy néu duei day) duge quy dinh trong céng thite cn dugc didu chinh dé 66 dugc méi trang én dinh va phi hop. pton va nuéc thit ho&e chat chiét nam men c6 thé thay di cde dac tinh dinh du6ng cia chung; thach cé thé thay d&I ste déng; cdc chat dem; mudi mat, chat chiét mat va deoxycolat, chét mau khang khudn, phy thugc vao cdc dac tinh chon loc. cia ching; = cht chi thi mau; — ec chat khéng sinh, phy thugc vao hoat tinh va twong tae voi cdc thanh phan khdc. CHU THICH: © quy mé céng nghiép, céc nha sin wut thirong dura ra obng thie da duge tbi wy héa dé dap Ung cae eu cht ‘ge hin. Théng thudng tre itn chon cfc thanh phn, sau d0 dibu chinh néng 40 gira cdc sén phim 48 thu duge eng higu néng v2 glém thidu bidn dng gitra cdc mé. 43.3 Nuvo BS chudn bi méi truéng nudi cy, chi st’ dung nude tinh khiét nhu nude cAt, d khi khodng, da loai ion hoac dug san xudt bang thém théu nguge hoc nude ob chét lugng tweng dueng khong chia céec chat gay tre ché hode anh hung dén sy phat tridn cla vi sinh vat trong cdc dig kin thir nghiém, Vi du: cdc vét clo, vét amoniac va cac vét ion kim loai. $6.CAS 18 Gon vi nhan dang Gon nn&t cla T8 chic Chemical Abstracts Service (CAS) d6i véi céc nguyén t6, oc hyp chét hoa hoe, palyme, cfc trinh ty sinh hee, c&c hn hyp va cic bye kim. 18TGVN 8128:2015 Nuc tinh khiét phai durge bao quén trong vat chira c6 nép day kin lam bang vat ligu trer (thy tinh ‘trung tinh, polyetylen v.v...) khéng chira ce chét gay we ché. Tuy nhién, sir dung nue mdi duo chung ct, $6 Iugng vi khuan khéng duoc vuret qué 10° don vj hinh thanh khuan lac (cfuy/ml va tét nat la dudi 10? cfu/ml, Sy nhiém khuan c&n dug kiém soat dinh ky theo ISO 6222", b Am & 22°C + 1 °C trong 68h 4h hode sir dung phwong phap turong deong. CHU THICH Nuée Joel in (68 Kh Khong) 06 thé cna krgng wi sinh vat rt cao, do d6, chi si dung toa! zo nay Khi ibm ta ham lngng vi sinh vt. Tham khdo ty via ota aha sn sauSt 68 tim Bign pha 2 nndtnhdengiden tid nhibm khudn. Nude 48 kniy khodng bi nhidm bln cao, tham ch 68 Khe tring bing cch lo, thi v8n of thé obn chéra cfc chét ma ob thd gay ‘re ché sy phat ttidn cia cac wi sinh vat nhdt dinh. D6 din dign cla nu6e duge sir dung trong phng thir nghgm khong duge lén hon 25 wS.em™ (tong duong véi dign tra > 0,04 MQ.cm) va tét nhat 8 derci 5 pS.cr” (nude dat chat Iureng loai 3 quy dinh trong TCVN 4854 (ISO 3696) ") & 25 °C, trir khi oé-quy dinh khac. £6 dln dién cla nuée can duge kiém tra trudec khi sir dyng. 43.4 Can va hoan nuoe ‘Tuan thi cdc canh bao va an toan, can clin than mot krong cAn thiét cla méi trréng khé ho&c c&c thanh phan ring Ié va khudy tron voi m6t lugng nude can thiét dé tranh vén cyc. Dung can chinh xéc 66 sai 36 t6i da cho phép nhd hon hoac bing 1% nu trong TCVN 6404 (ISO 7218) va TCVN 9716 (ISO 8199). Cac thanh phan phai dugc cho vo mét lvgng purée quy dinh, triv khi 6 quy dinh khac. 43.8 Hoa tan va phan tan ‘MOI trréng kno clin duoc phan tén nhanh bang cach khudy ngay va khudy lién tue sau 46 dun néng d& hoa tan, néu cn. Méi tréng chira thach cin durgc ngam trong nurée vai philt truréc khi lam néng, Khudy 48 hoa tan va saud6. phan phéi vao vat chiza_néu cdn, tnuréc-khi hp Ap lyre. Trénh 68 moi trdéng bj qué nhiét. 4.3.8 80 va chinh pH Dung may do pH 48 do dé pH va chinh truéec khi kh? tring, néu can, sao cho sau Khi khir tring va lam nguéi dén 25 °C, mdi trong 66 pH dat yéu cau + 0,2 don vi pH, trir khi o6 quy dinh khde. Théng thong sir dung dung dich natri hydroxit khoang 40 gil [o(NaOH) = 1 mol/l} hodc axit clohydric loang Khoang 36,5 gl [c(HCI) = 1 mol/l] 48 didu chinh pH. Vide dau chinn nay duye thye hién sau khi khiy tring, str dung dung dich 3 khir tring. Vigc chinh pH durgc néu trong TCVN 6404 (ISO 7218) va TCVN 9716 (ISO 8199). CHU THICH Moi tréng 06 ban sn thuémg cho théy thay 06% pH trrdc v8 sau khi hdp ap ire khde nhau G&ng k8. Tuy nin, bu sir dung nurbe cAt hoje nurée di khit khong, th vigc chinh pH tnréc ki hdp dp tye ka khong cd thigt, 19TCVN 8128:2015 43.7 Phan phéi Phan phéi mai trurbng vao cdc vat chia thich hep, dm bao cé khodng tréng phfa trén dé tranh qua soi trong qua trinh lam nguéi sau Khi xtr ly nhiét bang hp dp ie hoe lam tan chay lai hod lam tran sau khi thém cae thanh phan khdc. CHO THICH: Khodna tring nay 06 thd khong cB tht nbu ap vc trong nai hap dure duy trong subt qu ria lm nau 43.8. Khir tring 4.3.8.1 Yéu clu chung Khir tring méi truéng nubi cdy da chudn bj trong ngay sir dung. Khir tring méi trvéng nudi cdy va thuéc thir bing khiy tring wét bang nhiét (4.3.7.2) ho&e bang cach loc (4.3.7.3). Mét 6 méi truéng nhat dinh khéng can hdp ap Ive nhung ca thé dug sty dung sau khi dun s6i. Vi du: trréng Enterobacieriaceae chira xanh briliant thurong 6c biét nhay véi nhigt va anh sang va can ugc lam nguéi nhanh sau khi dun s6i va bao vé trdnh anh sang manh. Cling tuong ty, mét sé thude thir c6 thé duge siv dung ma khong cn phai khir tring. Trong moi trrgng hop, xem tiéu chuan thich hyp cy thé ho&e huéng dan cla nha san xuat. 4.3.8.2 Khir trong bang nhigt dm Vige khir tring bang nhiét 4m duc the hién trong ndi hap ap lc hoc may chun bj méi trvéng. D6i v6i cdc vat chira mol trréng voi thé tich lon hon 1 000 mi, chdp nhan chu ky Khir tring cia ndi hap 4p lye 48 dam bao xir ly nhiét day 42, Trong moi truréng hop, tuan thi cdc huong dan néu trong tiéu chudn cy thé hodic theo huéng dan cia nha san xudt. CHU THICH vige qué nhigt eye bp o thé xudt hign ki hp 6p lye moi trxdng v6i thé tien lm (> 1 000 mi. ‘Sau khi gia nhigt cn lam nguéi moi trvdng tranh dé trdo. Didu nay dc biét quan trong déi voi mdi trudng cé thé tich Ion va d6i vei cdc méi trvdng c6 cdc thanh phan nhay cam voi nhiét, vi du: mdi trudng chiva lye sing, ‘Théng tin bé sung vé Khir triing bang nhigt 4m duge néu trong TCVN 6404 (ISO 7218) [11]. \Viée khir trung bang nhiét can duoc danh gié sir dung eae gid trj Fo, 06 tinh dén vige xis ly nhiét trong subt qué trinh gia nhiét va lam nguéi. Vie xir iy nhiét c&n dure xéo dinh cho ting mé cy thé, d& dam bao xir If thich hyp c&c vat chira & bat ky vi tri nao trong ndi hap dp Ive. 20TCVN 8128:2015 4.3.8.3. Khir tring bang céch loc Khér tring bling cach Igc cé thé duge thyc hign trong cdc didu kign chan khéng hodc gidm dp sudt. Sir dung thiét bi va cdc mang loc vé triing 66 o& 18 0,2 um. Khir tring cdc b§ phan kha nhau cila thiét bi loc theo TCVN 6404 (ISO 7218) ho&c TCVN 9716 (ISO 8199) hoc sir dung dung cy 48 khi trang tude, ‘Mét sé mang loc 06 thé gil’ Iai c&c protein hoac c&c chdt khde (nhu khang sinh). © thu duc ndng 46 ding, agueti sir dying cdn chen kiéu mang lgc thich hyp, vi dy: mang lién két protein th4p va bé loc 44 Serge lam wit so bd. 4.3.84 Chuan bj cac chdt bé sung CANH BAO— Céc chit bé sung 6 chia céc cht déc, dic bigt Id céc chdt khang sinh, cdn duge aor ly cn than dé tranh phan tan vao bét cé thd gay dj dong hoge c4c phan tng cho nhan vién phdng thir nghigm. Thy hign cdc bign phép phang ngiva thich hyp va tuan thd céc huéng din = cirarnbdesarrxudt khi pha ché-céc dung dich. Khing sir dyng cdc ch&t bd sung durge pha ché a8 hét han str dung, d6i voi cc dung dich lam vic ia cc chat khang sinh chi sir dung trong ngay chudin bi. Trong nhtng tinh huéng nhét dinh, cc dung {ich Ichéng sinh 6 thd duge bdo quan dng lanh véi cc lugng thich hyp nhung khéng 4p déng lai ‘sau-khi.r& d6ng. Kha nang giém hoat tinh do dong lanh cdn duge nha san xudt thiét lap hose duge ngudi sir dung kiém tra. 44 Bao quan va han st? dyng méi trréng da chudn bj 444 Méltnréng sir dung ngay ¢é ban san tén thi trong Tuan thd cdc huréng dln cla nha san xudt vd cdc didu kién bao quan, han sir dung va cach sir dung. 44.2 M6ltreéng duge-chudn bj trong phdng thir nghigm 44.24 Yéu cdu chung Nhan biét tt cd cdc mdi trong 68 dam bao truy xu&t durge ngudn gébc. (Cac méi tring 06 thdi han sir dyng [a khde nhau. Trong cc tiéu chudn riéng 88 quy dinh cdc didu én cu thé varthei han si dung cia tong loai moi trong nhung phang thir nghiém phai thm ainh cac idu kign 66. Phong thir nghiém phai quy dinh tan sudt kidm tra x4c nhan. Bao quan méi turéng & cac didu kién khéng lam thay déi thanh phan mdi trvéng, trénh anh sang va ‘rénh bj khé. Bao quan & 5 °C + 3 °C, néu khong st dung ngay hoc theo quy dinh khéc trong tidu chudn cy thé. 21TCVN 8128:2015 Néu duigc bdo quan lanh, thi khong nén bao quan qué 2 tuan dén 4 tuan di véi dia va déi voi cae ng hode lo nhé thi Khéng qua 3 théng dén 6 thang, trix khi <6 quy dinh trong cao tiau chudn cu thé ho&c cc két qua kiém tra xc nhan thdi han sé dung ola phang thir nghiém cho théy lau hon. Théng tin 0d sung vé thei han bao quan téi da déi voi méi trréng pha ché san dugc néu trong TCVN 9716 (ISO 8199), Tai ligu tham khdo [17] va [21] Khuyén cdo, mai truéng dug bd sung c&e thanh phn khéng én dinh thi can dugc str dung trong ngay, trir khi c6 quy dinh trong cc tiéu chudn cy thd hoc cdc két qua kiém tra xdc nhan théi han sir ‘dung cia phéng thir nghiém cho thdy au hom (4.4.2.2), Mai trréng dac 6 chiva cdc chat khong 6n dinh vafhoac cc chat hoat hoa dé tan chay lai khong nén bao quan voi krgng lon ‘Trude khi st? dung hoa tréc khi lam néng tiép, nén can bang nhiét 68 mdi truréng nudi cdy dén nhiét 46 mai trréng xung quanh, 4.4.2.2. Danh gia han si? dung Han si? dung 461 véi mdi trréng nudi cdy 44 durge bao quiin phai durye thiét lap bang cach kiém tra mi truéng sau khong théi gian bo quan xéc dinh vé cdc 6c tinh vat ly, hda hgc va vi sinh vat theo ‘quy dinh trong tiéu chuan nay. Phong thir nghiém phai quy dinh tan xuat kiém tra x4e nhan. Kiém tra sy thay di bat ky vd mau so, déu higu bay hoifmat nude, thay d3i pH ho&c higu sudt hoac tinh chon loc va tinh dc higu khi cé thé. Han str dung phai dya vao théi gian bdo quan ma mdi trong vin gitr duge cae tinh ning quy dinh CHU THICH: Vige kiém tra nay cfing thich hop khi thir kiém tra cdc moi truéng ban s3n. 4.4.2.3. Bao quan méi truréng trong dia Petri ‘Sir dung ngay mdi trong déng ac hodc bao quan trong eéc didu kién khéng lam thanh phan cia mdi truéng bj bién déi va mat nude, nghia la 48 o-nettérvamnose-trong tir ianh & § °C + 3 °C. Dan nhan trén day hodc mat bén clia dia dién ngay thang chudn bi vayhoac han sir dung va cach nh4n biét. Cach khae, c6 thé str dung cdc hé théng ma hoa dap tng diroe cae yeu cdu nay. Han sir dung cla céc dia thach sé tang lén néu duge b3o quan trong cdc tii bang chat déo hose cellophan han kin. B& gidm thiéu ngung ty nue, thi can lam mat cdc dia truée khi cho vao tii. Khong lam khé bé mat céc dia thach truéc khi bao quan lanh. 45 Chudn bj dé sir dung 4.5.1 Lam tan chay méi treéng thach nudi cdy Lam tan chay mai truéng nudi cAy bing céch a8t trong ndi cach thuy hodc bang cach khac cho két qua twong ty [vi dy: dong hoi qua hap ép lye nhur quy dinh trong TCVN 6404 (ISO 7218) ho&c TCVN 9716 22TCVN 8128:2015 (ISO 8189)], Méi trrdng tnrdc dé da durge hdp 4p Ie cdn duge lam néng lal trong mét thdi gian t6i thidu a8 duy tri cht lugng méi trudng. Trénh qua nhiét va khi d& tan chay thi ly mi trwong ra. BE & nhiét d6 phéng trong khoang thai gian ngén, vi dy: 2 min, tre khi dat vdo ndi cach thy 48 am nguéi, khéng lam thiyy tinh bj v&. CAn ni léng nap day truéc khi gia nhiét va van chat tric khi dua ra khdi ngudn nhiét. Lam ngudi moi trrémg tan chy duy tri tir 47 °C dén 50 °C trong nbi céch thuy kiém soat duoc nhiet a9. Thoi gian thy 18 on thiét 48 dat dug 47 °C dén 50 °C phy thugc vao loai mdi truéng, thé tich va sé lueng don vi 48 trong nbi c&ch thuy. Méi trréng & tan chay thi si dung cang sém cang tét, nhung khéng nén 48 qué 4h. Trrdng hyp mai trang dc biét nhay cém, thi thai gian gidr méi trréng tan chay phai ng&n hon, theo quy éinh trong tiéu chun cy thd lién quan. Méi truréng tan chay chua sir dung khéng durge lam é6ng dc lai 48 sir dung cho Ian sau. ‘Thiét 1p va ghi thanh van ban ché d9 didu chinh thach bang cach cai dat nhiét ké trong méi truéng thach trong hOp chtra riéng tuong ty nhu dugs sir dung cho mdi trréng thir nghiém. Biéu nay phy thude vao sé lvong va kich thuéc vat chira dat vao nbi cach thy. CHU THICH: Mbitrréng éizge sit dung trong phuong phép 48 dia, duryc bb sung vo mau cin durge chinh dén 44 °C dn 147 °C, hobe theo guy dinh trong tiéu chun Hen quan. Sir dung ndi c&ch thiy cdi d&t & 44 °C dén 47 °C. Thing tin BB sung vb str dng va kidm tra nbi cach thay dura nu trong TCVN 6404 (ISO 7218). 4.6.2, Logi khi mél truéng nudi cdy D8 cb ham lugng chinh xac khéng khi/oxy, néu cn, ngay tneéc khi stv dung, lam néng mdi trrdng trong ndi cach thuy dun séi ho&c duéi dong hoi nurée 15 min, noi lng np; sau khi lam néng, van chat ndp va lam ngudi nhanh dén nhiét 46 lam viée. 48.3 Thém chat bé sung Cac thanh phar khéng ban nhiét cdn duge b3 sung vao méi trvdng sau Khi 34 lam ngudi dén dudi 50 °C. Néu méi trirémg chiza thach, 48 cho chat bé sung v6 tring cén bling dén nhiét 6 phéng trréc hi b3 sung vao méi trréng thach. Viée thém cdc chdt bd sung lanh dang ling 6 thé lam cho thach dong lai hoc tao thanh cdc lép vay trong sudt lam cho phan bé khéng dung, Gan thyc hién theo ding huéng dan ca nha san xudt. Nhe nhang trén ky tdt cd cdc thanh phan vao méi tréng va phan phéi vao cdc hép chira cudi cing cang nhanh cang tét. 48.4 Chudn bj méi tnréng diac trong cae dia Petri R6t mai trrdng thach nudi cdy 43 tan chdy vao cdc dia Petri sao cho thu dug mot lop day it nhét ta 3mm (di véi cdc dia c6 dudng kinh 90 mrh, 18 ml dén 20 mi thach thurng 4 4) hoc theo quy dinh trong tiu chudn lién quan. Néu cdc dia duge bao quan hode néu G 4m qua 72 h, hoae nhiét do a tren 40°C, thi cin dén mét lung mdi truéng nuéi cdy lon hon, 88 thach ngudi va déng dc lai bing céch dat céc dia cling vi nap trén mat phing n&ém ngang, & noi mat.TCVN 8128:2015 Cac dia méi trvéng nudi cdy 8 chuan bi sfin 48 sir dung ngay cdn duge bao quan va sir dung theo hung dln oa nha sdn xudt. 4.5.5 Chudn bj méi trang 4d dia dé nudi cay 8 nudi cdy bs mat méi trzdng dc, thi cAn lam khd nhanh cdc dia ngay tree khi sir dyng, cho dn khi khong cén céc giot née trén b8 mat mdi trvéng, Khéng lam qué khé bé mat thach, ‘Dé iam khé cae ala, cdc diém sau day rdt quan trong: = 88 dm ofa méi trang nudi cdy 1a quan trong vi viéc phat trién t6i uu vi khudn phy thude vao 4c. Giéu kién am trong va trén méi trudng. Vi du viée that thot qué nhidu nude ¢6 thé dan aén tang néng 8 cdc chdt-gay We ché trong mi trvéng nudi c4y chon loc va giam hoat 46 nude trén be mat moi ‘trueng. = hi vi khudn durge nuéi cy khong lan nhanh va cae dia khé do thd’ tiét thi khéng can phai lam khé. Trong trréng hep nay, cé thé bé qua quy trinh sy vi chi lam tang kha nang nhiém ban va that thoat muroe khéng can thiét, — Chon nhiét 46 va thoi gian lam khé sao cho méi truéng cang it bj nhiém ban cang tét va vige gia nhiét khéng nh hung d&n chét lugng méi trxong nudi cdy. Thoi gian lam kh8 phy thuge vio mirc 6} ngung ty nuréc trong dia Petri, nhung phai cang ngan cang tét. — 88 trénh nhim ban va néu cc dia khéng lim khé trong ti sy thi céc dia phai duge lam khé bang cach Up nguge dia voi b& mat méi truéng nudi cay at Up. Trong thyc t8, cac dia cé thé dugc lam khé bang céch dat dia voi bé mat thach lat dp va md n&p mot nik va dé trong td & nhiét dO tir 25 °C dén 50 °C. Lam khé cAc dia cho dén khi hét han cdc gigt nude trén b& m&t nap. Khdng lam khé thém. Cac dia thach efing c6 thé dug lam khd voi bé mat thach huéng-lerrtrén dé trong tl céy ( nhigt-66 phdng) tir 30 min Gén 60 min hoc day ndp dé qua d8m & nhiét 6 phang. 4.6 U 4m méi trvréng dac trong cac dia Petri Trong qua tinh & 4m, méi truéng thach sé mdt nude. Trong mét sé trudng hgp, diéu nay ¢6 thé anh huéng dén su phat trién cia cdc vi sinh vat. Cac yéu té dnh huréng dén sy mat nuée Ia thanh phan “néi tniréing. iomg méi trudng trong da, kiéu ti 4m ¢6 quat hoc kiéu khdc, 46 Am ciia khong Khi trong ta 4m, vj tri va s6 lugng dia trong ti 4m va nhiét 46 G. Co thé gidm thidu vige mat nude bang cach chéng sau die dat vao tui chat déo mé miéng phia trén (4 tranh ngung ty nhiéu nude). Cach khao, 6 thé lam tang 6é am cia khéng khi trong ti 4m bang cach dat hép dng nuéc & phia duoi. Thuong xuyén thay nude dé trénh nhigm ndm méc. 24TCVN 8128:2015 4.7 Thai bé méi trong ‘Thai bd méi trong bj nhiém ban va khdng str dyng theo cach an toan va dap teng duge cdc quy dinh hign hanh. 5 Visinh vat thi nghiém dé thir higu nang 5.4 Yéucdu chung Cac téu chudn méi ho&e G8 duvge soat xét pha quy dinh r& phép thir higu nang cia méi trréng nusi ody, bao gém quy dinh v8 cdc ching kiém ching va cac tiéu chi chap nhan, theo Phy hye J. 5.2 Chon visinh vat thie nghigm 86 visinh vat thir nghiém c&n c6 cac vi sinh vat 6 cdc dc tinh én dinh dai dién cho cc lai va o6 thé tin cay 66i véi viée thd hign higu ning 161 uu eda méi trrdng duoc ohong thir nahiém chudrrbi cy thé. Cac vi sinh vat thir nghiém gdm cdc chiing cé sn trong cdc b@ suru ip giag Goi chung, hung cling 6 thé bao gdm cae chiing dc trung do phang tht nghiém phan lap. Cac dac trumg nudi cay fén quan cia chiing géc di chtrng cn durgc kiém tra va durge phéng thir nghiém ghi lai hodc chon ching moi néu xuat hién cdc diac trung khéng didn hinh. Tét nhdt la slr dyng ode ching od ngudn géc tir hye phdm hod tiv nude cho du khéng phai tét cd cdc bé suru tap céc ching nudi ody du cung cp cdc dr Iigu 46 v8 ngudn gée xudt x cba chting. Cac dac trung v8 giéng cla ge déi chimg lién quan phal duge phéng thir nghiém kiém tra va ghi lal. Néu 06 sv bién déi vé chiing, tht nghién ctu cde kha nang anh hudng cia méi trong nudi cdy bang cdch thu thap cling mt méi trrgng tly cdc nha sdn xudt khdc nhau va thu Wy c&c gidng déi-chimg bd ‘sung tir b6 suu tap giéng. LUU ¥: Ngui sir dung can phan hdi théng tin lién quan dén cac bién abi vé ching va higu nang. Cae vi sinh vat thir nghiém 46i voi mi méi trrong 06 thé bao gdm: = cde ching durong tinh manh véi céc dc rung dién hinh ciia vi sinh vat dich; cc ching durong tinh yéu; = cde chiing &m tinh khéng cho théy céc dc tinh mong di alla vi sinh vat dich (ede de trung &m tinh); = céc chiing bj irc ché mat phan hodic hoan toan. Phy luc E dua ra cdc vi sinh vat thir nghigm Gurgc str dyng trong céc fiéu chun vi sinh vat trong thyre pham va Phy luc F dua ra céc vi sinh vat thir nghiém due sir dung trong cc titu chun vi sinh vat trong nude, 25TCVN 8128:2015 CHO THICH Mot vai han ché oie quée gia va mét wai huéng dn yeu cdu si dung cdo typ huyAt thanh khac voi cac tye truyét thanh quy dinh trong oc Béng nay. Néu 1 cic yu cdu ca quéc gia lian quan dn vige chon céc typ huyét than Salmonola 5.3 Bao quan va duy tri cac vi sinh vt thir nghiém 6.3.1. Yéu cdu chung Hign nay c6 sn mét vai phuong phép béo quan va duy tri thanh c6ng tt ¢& céc vi sinh vat lidn quan én vi sinh vat trong thye ph4m va trong nue, vi dy: déng khd, bao quan cac hat & U 70 °C, hoac sir dung nito lang. Mét phuong phap 6 thé khéng thich hyp cho tat ca cdc ching. Cac phuong phap bd sung 48 bao quan cc vi sinh vet néu trong [14], [15], {36], [37] va [38]. $4 long vi sinh vat thir nghiém duge c4y chuy&n c&n duge lap thanh van ban dé ngan ngtva cdy chuyén qué nhidu lam t&ng nguy co bién ddi kiéu hin, Mot lin cdy chuy&n dug xéc dinh la chuyén tle ging vi sinh vat séng sang mdi trong méi voi sy phat trién olia cdc vi sinh vat. Moi kiéu cdy chuyén dugc coi la dang cAy chuy&n/chuyén sang, Thong tin bd sung o6 trong [27], (28), [35], (38). Xem so 6d trong B.1, B.2 va Phu luc B vd thong tin bd sung 48 duy tri va chuan bj. 5.3.2 Vi sinh vét thir nghiém tir cdc nguén thurong mai Néu cc vi sin vat thir nghiém thu dugc tir cdc bd suu tap chudn hodic cae nha cung c&p duge cp chimng cht ISO 9001[2] hoc chieng nhgn thich hep khdc va duge gift trong cae hp dyeng géc, thi phai sir dung va nudi ody theo huéng dan cia nha san xual, Phang thir nghiém c&n che chan raing ching dugc cung cap la ching déi chieng ho&c géc déi cheng va bao nhiéu lan cy chuyén 68 dug thy hién trvée khi nhan va tai igu vé théng tin lién quan. Phdng thir nghiém can kiém tra sy c6 mat cdc dae tinh mong di. 5.3.3 Ching géc 461 ching dure chudn bj trong phdng thir nghigm Cae giéng géc cla cdc ching a6i chieng (xem Hinh B.1) diing cho cdc myc dich thir higu nang phai urge duy tri va xir I theo céch sao cho gidm thiéu kha néing nhi&m chéo, bién adi hoc thay déi cae tinh chat dién hinh. Cac géc di chteng can duge bao quan véi céc iugng nhd, thudng duge duy tri & nhigt d@ dong tanh sau (vi du, du6i — 70 °C) hodc déng khd. 0 nhiét d> cao hon thi théi gian séng c6 thé gidm va oo sy bién didi vé di truyén. (Cac d&c trung phat trién ciia chung can duge lap thanh van ban day di cho tirng mai truéng ma trénitrong 46 cdc ching nay dug sir dung lm sinh vat thir nghiémn, Cac abe dbi chieng khéng du¢c ding 48 chudn bi cdc ching aéi ching. 26TCVN 8128:2015 5.3.4 Gidng géc Cy gidng gb thuéng duge chulin bi tir gbc déi ching dong sau ho&e Géng khé (xem Hinh B.2). Cac liwgng nhd phai durge xir ly theo cdch sao cho tranh duge nhi8m b&n chéo géc abi ching vashoge suy gidm cht lugng cila ching. Cac giéng géc cin dirgc chun bj bang cach hoa lai mét lugng géc 461 cchizng trong hode trén mei truréng phat trién khéng chon loc va U 48 tao ra giéng pha tinh. Xem 5.3.2 vicacySe cau bao quan va hd so uu tri 186i v6i cdc he théng bao quan cé ban sn, can tuén thi nghiém nga cac huréng dan ota nha edn xudt C4c giéng gc khong duoc ding 68 chudn bi cdc ching a6i ching hoac géc 464i chirng. 5.3.5 Giéngtemvige Céc ging kam viéc phai-durre chuan bi tir giéng géc ho&e abc a4 ching va duge st dung a8 chuan bj gi6ng thir nginieen. Cc gidng kim vigc knrong curve dimg 48 chuan bj cde chiing 66i ching, g6c 461 chteng hode giéng gbc hoac tao ra giéng kant wigc fiép theo. 5.4. Visinh vot ad thir higu nang 6.4.1 Yéucdu chang (Cc vi sinh vat thich hop 48 thir higu nding théng thuréng dug néu trong cdc Phy luc E va F. (Cc thé tich dich ody va sé rong vi sinh vat dug sir dung, xem 6.4.2.4 va 5.4.2.5, Huong din duéi day ta mét vi dy vé quy trinh thich hop 48 tao ra dich ody tiéu chudn 4 kim soat chét lvong mdi trurémg. Cac auy trinh nay dec ép dung trong trréng.hop chung nhung mét sb.sinh vat 06 thé yéu cau cac didu kign chudin bj dac biét, vi dy: cdc sinh vat ki khi, va man, ua thdm théu va cdc vi sinh vat 66 yéu clu phat triért hoc dinh dung dic biét. 8.4.2 Chun bj 5.4.2.1 Chuan bj giéng géc Khi yéu cau cdy gée a6i ching vao méi trréng diac, vi dy thach trypton dau tueng (TSA) hoc TSA mau, cy sao cho thu duge céc khudin lac moc don |8. U trong cae didu kién thich hyp, vi dy: ode vi khudn hiéu khi & 37 °C trong khoang air 78 h dén 24 h. Kiém tra giéng géc trén méi truréng 4c vd 6 thuan khiét va sir dung trong khodng thdi gian quy éinh (vi dy: 14 ngay & nhiét 4 thich hop 68 trénh lam thay d3I ding ké vi sinh vat). a7‘TCVN 8128:2045 5.4.2.2 Chuan bj giéng lam vige Céc giéng lam vie phai durge chudn bj tir gc déi ching (ho&e khi cn thi tir giéng géc) nh ‘gidng pha ‘inh thuan kiét trong mi trréng canh thang khong chon loc. Oi véi nhieu vi khudn hidu khi thyrng thy duge bang cach ti Am 18 h dén 24 h. Gidng lam vige 06 thé dug chuan ®/ tir chdt chudn thurong mai RM hac CRM hod dugc chuan bj trong phéng thir nghiém. Nang d@ tia huyén phd dug chuan bj phai én dinh va dong nhét trong suét qué trinh si dung (7), (10), [11], [21], [29] va [30} C6 thé sir dung cdc kj thuat khac nhau, nhung phai dm bdo d6 thudn olla dich cy, cing nhu suv chudn héa 68 cho phép sir dung vé sau. Tay thude vao kich thuée khuan lac, dy mét dén hai khudn lac tir mdi trut’ng giéng g6c bang vong cy. Nén siv dung ving cy 1 ul dé tranh ly qué nhidu dich cay. Chuyén dich ey ln méi tru’ng long khéng chon Igo, vi dy: canh thang trypton Sau trong (TSB) va tron kj trén may ten vortex. U Am trong céc digu kién va thai gian thich hep (vi dy: 6 véi phan Ién cc vi khudn hiéu khi thudng o 6 37 °C trong 18 h dén 24h). Sir dung gidng lam vige nay trong khong thoi gian quy inh (vi dy: téi da ba ngay & nhigt 46 thich hop 8 tranh lam thay déi dang ké vi sinh vat). ‘Chuan bi va bao quan cde bao tir ndm va vi Khudn ding lm giéng lam viée. xem {10}, £11], [24], [25] va (30), 5.4.2.3 Chuan bj céc huyén pha (dich cdy) 48 thir nghigm Chun bj cde day dung dich pha long trong cdc dich pha loding thich hgp (vi dy: dung:dich Ringer ndng a9 %, dung dich mudi pepton) va st dung buéc pha loang thich hyp nhat 46i voi sé fureng vi sinh ‘vat mong muén (cfu) trong mét thé tich quy dinh. Dung dich pha loding thich hop dug sir dung lam dich ey thir nghiém cn dure xée dinh tir cdc phép thir true 66 trong cdc didu kién chudn héa nghiém ngat d6i voi tét c& cdc bude. ‘Sv dung cc huy&n phil (dich céy) trong théi gian quy dinh (vi dy: d&n 2h & nhiét 33 phdng hoa Tang 24 h néu duge bao quan & § °C + 3 °C hodc thdi gian bao quan lau hon néu 8 dupe Banh gié hi Ive [10], (21). Co thé si dung dich cdy dong lanh néu cho thay vi sinh vat cé thé s6ng trong thdi gian dure chon. 28‘TOVN 8128:2015 6.4.2.4 Thé tich dich cdy ‘Thé tich dich cy dure str dung cho phép thi higu nang dinh lwong phdi phan dnh dug cdc dich céy si? dung trong cde didu kign thir nghiém é6i voi méi trréng [én quan. ‘Ai voi dich pha long va mol trréng léng duoc sir dung cho phép thir dinh Iugng, thi thé tich dich cy phai 6 cling ty I véi thé tich dich c4y durge sir dung trong tiéu chuan li8n quan, thudng [a 10 % mai trudng can thi, 8.4.2.6 Mirc dich cay 5.4.2.6.1 Mie dich cdy 48 thir higu sudt 8.4.2.6.1.1 Phép thir dink lugng ‘B6i voi phép thi dinh ivgng, thi mirc & khoding 10? cfu la cn thiét d8 dat duge do chym day da (em Bang 1). Phép thir nay cé thé can si dung nhidu hon mét dia kép. Can sir dung dai thy t8 tir 60 cfu dén 120 cfu tran dia voi s6 dém t6i thidu 50 cfu trén dia. Ddi voi c&e b6 loc, 36 dém cfu turong tur can dén vie sir dung méthode nhiéu mang 9c. Bang 1 cho thay misc tin cay 95 % lién quan dén sé aém khuan lac, ‘B6i v6i cdc phép thir dinh Iuong cia dich pha lodng va méi truéng van chuyén lng, thi mize dich cy & khodng 10° an 10‘cfu la cdn thidt a8 dat dure dich cdy & khong 100 cfu trong thé tich duegc dan ‘trén dia. Bang 1 - Mirc tin cay 95 % d6i voi 36 lweng khudn lac gid djnh théng nhdt véi phan bé Polsson™1) 24 $6 lugng khuan ge dém dure Gidi han 49 chy GiGi han tin edy xdp x1 85 % (chinh xae dén phan tram) % 600 a8 465 dén 545 400 £10 360 dén 440 320 an 284 oén 356 200 #14 172 66n 228 100 £20 80.aén 120 80 £22 62 dén 98 Gd £28 36 dén 64 30 £37 19.4641 20 447 11 aén 29 16 £50 aén24 10 £60 406n16 6 £83 108011 29TCVN 8128:2015 6.4.2.5.1.2. Phép thir dinh tinh ‘Thé tich duge sir dung cho phép thr can chira: — 10° cfu dén 10* cfu cho cae phép thir dinh tinh moi trudng a8 dia; — $100 cfu cho cdc phép thir higu sudt cia céc méi tréng tang sinh va tang sinh so’ bd; = 10 cfu dén 10° cfu cho cdc phép thir dinh tinh mdi truxgng van chuyén ac. 5.4.2,5.2 Mirc vi sinh vat trong dich cay 46i voi phép thi? tinh chon loc ‘D6i voi cc phép thé tinh chon I9c eda méi truéng nudi cdy, nudi cy huyén phil vi sinh vat khéng phai dich c6 chira tir 10* ciu dén 10° cfu lén dia thach hoe vao éng dyng mai trveng. 5.4.2.5.3 Mire vi sinh vattrong dich cdy d6i voi phép thir tinh dac higu ‘6i v6i cdc phép thi dinh tinh cia méi trong 68 dia, vé tinh dac higu cn dén 10° cfu dén 10° cfu. 5.4.26 U U mét trong aa cdy thea cae didu kién quy dinh trong cc tigu chudn lién quan. Phy lye E néu cdc didu kién U due stv dyng trong cac tisu chudn quy dinh déi véi vi sinh vat trong thye phd va Phu luc F néu cc Gidu kign U durgc sir dung trong cdc tiéu chudn quy dinh a6i véi vi sinh vat trong nuée, Tranh th&t thoat née trong méi trréng thach trong qua trinh U, xem 4.6. Cn st? dung théi gian & ngdn nhét trong tiéu chuan duge diing d6i véi vi sinh vat dich, trong khi thoi gian Gi cho phép lau nh&t c&n duge sir dung khi x4c dinh tinh chon loc [10]. 6 Kiém sodt chéthrong-va-thir- higu nang cla méi trong nudi cdy 6.1 Yéu cau chung Cac diu sau day mé ta cdc yéu cau 46i véi tét cd cc mdi trudng nudi edy. Cac yéu cdu nay c6 thé ap dyng cho moi c& mé méi truréng nusi edy. Thue té, cc mau 06 thé chira cac vi sinh vat bj te ché. Tinh phi hgp cia mai truréng lién quan dén do, phyc héi cla cact&-bao bi re ché. cn deere tinh dén [21], [31], [32], [33]. Chat Ivong clia méi tring nuéi cdy phy thude vao chat lugng cla cdc thanh phan co ban, céng thir ding, cht lvong olla quy trinh chudn bi, vide loai trir nhiém khudn va cdc didu kién bao géi va bao quan. 30TGVN 8128:2015 Kiém soat chét long méi trxéng nudi cy phai his hop voi vige sir dung ciia mai trxeng (vi dy: dinh tinh hoge dinh lugng). Truée khi str dung, higu nang cla ting mé moi trong nudi cdy phai durgc thir nghiém phan loai méi trréng trong 6.4. Néu khéng thé thye hién phép thir true khi sir dung do tinh khéng én dinh cia méi trvréng hode bd sung, thi phai thyrc hign phép thir higu nang song song véi viée thir nghiém m&u. 6.2 Kidm soat chit lung ly -héa Moi trurbng nudi cy hoan chinh phai phi hop va céc tinh chat Iy-héa nh quy dinh trong tiéu chadn tuong img. Ngoai ra, viée danh gid chat iugng bang kiém tra cam quan phai dam bao ting méi trréng nudi cdy du 44p-tng durge cae yéu cau quy dinh, vi du: lwgng phan phéi vashode 4 day; — bd ngoai, mau sdc va tinh dng nhdt; 6 dong ac: = 49 dm; Ngoai ra, phai xc dinh tr sé pH. Céc than phan ring 16 va moi thanh phn bé sung chen loc hod thanh phan dinh duéng phai duee anh gia chdt lvgng bang cdc quy trinh thich hop. 6.3 Kiém soat cht luyng vi sinh vat 6.3.4 Yéucdu chung Viéc thyc hién c4c phép thir hiéu nang vé vi sinh phai duge tién hanh trén mu dai dién cia mé san phdm cudi cing (6}, [8, (91, [21] 6.3.2 Méi trurng a6i ching D8 c6 duge dé tin cay cia cdc két quad phép thir higu nang, thi phai sir dung mi trong déi ching cb chét lvgng phi hyp cao. Cac vi dy can duge ngudi sv dung xem xét nhu sau: — sit dyng RM dinh long (xem 3.4.6) c6 chia mat lugng xéc dinh céc vi sinh vat khi danh gid mal trong a6i ching; — sit dung qua trinh sn xudt xéc inh ké ca viée lam tan chay, néu cd thé: — st dung cling mét co s& san xudtinguén géc moi truémg/cdc thanh phan; 31TCVN 8128:2015 — st dung mét lugng Ién vi sinh vat khi thir kidm tra (88 bao trum dai vi sinh vat cn tim); — chon “mai trzéng dbi ching’ cho myc dich d&nh gid; = cde quy trinh thich hep 48 dm bao chAt long khi sir dung méi trong abi ching. Cé thé khéng cAn thiét phai bao gdm tét cd cdc ndi dung trén khi danh gia sy phi hop cia méi trréng bi chizng. Phang this nghiém phai ty chon quy trinh phit hgp cho minh. Céc vi sinh vat thi nghiém phil hyp, phuong phap kiém soat va cdc tiéu chi ch4p nhan 461 voi mdi trwéng di ching thach trypton dau tong (TSA) dure néu trong céc Phy lye E va F. Cé thé sir dung cdc méi tréng 46i ching khong chon loc khac néu Gap tng duge cc tiéu chi trén. 6.3.3 Kiém tra mire 46 nhiém vi khudn (kiém tra 46 vé tring) Can phai kiém tra mirc 49 nhi&m vi khuan (46 v6 triing) ctia mdt Ivgng médi trudng nudi c4y thich hop, tiy thuge vao o& cilia mé bang cach d 4m & cae didu kién thich hop. Cac mau thir nghiém phai it nhdt 1 mot dia hoac mot dng cho cde mé nhé (< 100 don vi). DSi voi cae mé Ién hon, ngudi sn xudt phai ldy theo quy dinh, vi du: dia vao cdc thanh phan méi trvdng, thong 96 qué trinh, céc giéi han va kidu bao géi, siy dung céc giéi han chét Ing diye chp nhan. Thong tin b} sung duge néu trong TCVN 7790-1 (ISO 2859-1) [6] ngoai cc tidu chudn quéc gia khac va c&c ngudn khéc {9}, {21} Céc tigu chi chp nhan phai duge thiét lap va diu chinh 46i v6i tizng mi trvdng. 6.4 Yéu cau chung 46i vi phép thir higu nang vi sinh vat 6.4.1 Yéu cau chung é anh gid mé méi truéng nudi c4y hoan chinh, thanh phan dinh duéng hodc cac chat bd sung, sy phat trién phai dug d4nh gia thich hyp bling c4c phuong phdp dinh tinh ho&e éinh Ivong néu trong tiéu chudn nay. ‘Si dung thiét bj thich hop va thy hign ky thuat nudi cy néu trong céc tiéu chudn lién quan, nudi cdy cdc méi truéng nudi cdy long ho&c ban dc vei mét lugng thich hgp (5.4.2.4) gidng lam viéc cla méi vi sinh vat thie nghigm xéc dinh, xem Phy tuo E va Phy lyc F. Cc vi dy vé phuong ph4p thé dinh tinh va dinh lugng déi vai méi trvdng nudi cy dc va Iéng duge néu trong tiéu chun nay. C6 thé chon phurong phdp bat ky khdc va khdng phai sir dyng t&t cd cdc phuong phap. Khi méi trong nudi cdy due sir dung 48 aém vi sinh vat, phai thre hign cdc phurong phép thir dinh lugng. 32‘TCVN 8128:2015 Khi c&n anh gié méi truéng nudi céy méi hoe nha sn xuat méi, nén thc hién cdc phyong phap thir Ginh long 48 cung cp théng tin bd sung v8 sy thay adi. Trong méi trudng léng, cdc tong tac din dn sy phat trién cdc vi sinh vat rét phirc tap, do d6, céc phuong phép thir higu nang cling phic tap hon di voi phép tht? nghiém cdc hén hyp méi trvdng dc bang cdc bd loc mang, sir dung ISO 7704. ‘Théng thirdng céc ky thuat vi sinh i gid dinh va-do 46, cdc phurong phdp khong diva ra chi tiét ey thé. Cac vi sinh vat thir nghiém thich hgp, cdc phurong phap kiém sodt va céc tiéu chi chp nhan duc ligt ké trong Phy luc E va Phy lye F. Tan sudt thir nghiém phai do ngubisir dung diéu chinh, 6 tinh één pham vi chudn bj trong phong thi ghifm cia ngwai si dung cubi cling va mirc dam bao chat ivong tai dé 6.4.2 Méi trudng sir dyng ngay Cac nha san xudt méi trvéng str dung ngay cé ban sin tn thi trong, dac bigt néu g4 duge ching nbn phi hop voi TCVN ISO 9001, phi od sn chereng trinh chat lugng va ¢é thé kam theo ching chi va chat lugng cia mdi trréng ma ho cung ep. Trong cdc didu kién nay, nga sth dyng cé thé khdng cAn phai thir nghiém thém vé mdi truéng 46 nhung cAn dim bdo ring cdc diéu kién bao quan duge duy trl theo huéng d&n ca nha san xudt. Déi véi méi truéng hoan chinh siv dyng ngay ¢6 cht bs sung, chét lvgng 44 durge nha san xudt kiém ‘so4t phu hgp vai tiéu chudn quy dinh, thi it nh&t nén c6 phép thir éinh tinh. Ngwi sir dung phai chéc chan rang nha san xudt cdc méi trréng dé sir dyng ngay c6 ban sn trén thi trong déu c6 chong trinh chét lugng d6i véi ce san phdm nay va cdc ching chi kiém soat chdt long dap Ug cc yéu edu oiia tiéu chudn nay, thda. man cdc két. qua dy kién va két qua-thu duge. Phong thir nghiém sir dung cling phai kiém tra cdc tai figu bling chimg a4 dam bao rang cdc tiéu chi ch4p nhan cia nha sin xuat vé phép thir higu ning dap img-cac yeu chu ndi bd cia phéng thir nghigm. Phai dinh ky kidm tra d8 ching minh chat lugng cia méi trvéng dug duy trl trong qué trinh van chuyén. Cting can phai kiém tra sau bdo quan va xi ly tiép trong phong thir nghiém, vi dy: sy tan chay cia moi truéng. Bigu chinh tan xudt kiém tra. 6 v6i mai tru’ng Khong hoan chinh cn duge phong thir nghiém thém céc chét bb sung (xem 3.3.5.1), thi kiém tra vige bé sung bang cdc hd so hoa thy hién cde phép thir dinh tinh 8 6m béo rang chat bd sung dug thém dung. 33‘TCVN 8128:2015 6.4.3. Méi trvéng due chudn bj tir cac céng thirc thanh phan khé ban san trén thj trudng ‘6i voi méi trvdng 6é dém vi sinh vat, thi phai thu hién phép thir dinh lugng, O6i voi cdc mai trong Khe thi chi can phép thir dinh tinh la da. Cac phép thi dinh lugng cho 4 dam bao chat lugng mdi trudng cao hon. ‘B6i voi cdc mai trong khéng quy dinh trong Phy lye E va Phy luc F thi viéc kiém soat chat long cin quy dinh theo huéng dn cla nha san xuat. 61 v6i cde méi truréng khéng chiva cdc chét chi thj hog cdc chat chon lgc, thl sv dung mét sé lugng han ché chiing thir nghiém. Bi v6i cdc méi trvdng 66 chiza cdc chdt chi thj ho&e ch4t chon Ic, thi sir dyng cdc chiing chi thi hoe chon Ioc. B4i véi méi trréng phire hyp, nghfa la cé cht bd sung thi mé cdn duge kiém tra véi cdc chiing 06 cdc dac tung duge liét ké trong 5.2. 6.4.4 Mdi trrdng dug chudn bj ti” céc thanh phan co’ ban ring ré Ngoai cae yéu cdu néu trong 6.4.3, thi can tién hanh phép thir dinh long dé kiém tra chidu hudng chét lugng cia vat ligu co ban, higu sudt cla méi trvdng va cdc quy trinh chudn bj sn xudt trong hong thir nghiém. 6.5 Danh gid higu nang va didn giai cdc két qua M@ méi truéng nudi cdy cho th4y d4p Gng néu tit ca cdc vi sinh vat thir nghiém dug sir dung thyc hign theo cdc quy dinh ky thuat. Méi trvéng duge ch4p nhan néu ca tiéu chi chung lan cac tiéu chi chat lugng dugc dap tng. Néu khong dap tng yéu cdu vé higu nang, xem Phy luc H. 6.6 Méi truéng khang djnh va thudc the 6.6.1. Méi tréng khiing djnh Higu nang cla méi trréng nudi cdy du¢e six dung cho cae phép thir khang dinh phai dugc x4c nhan truéc khi sir dung. Cac vi sinh vat thir nghiém am tinh va duong tinh thich hgp phai dug sir dung 48 x4¢ nh§n theo cach tong ty trong cdc tiéu chun cy the [9], [16]. 6.6.2 Thudc thir khang dinh Higu nang ota cée dung dich nhudm Gram, cée thuéc thir nhur Kovac, VP, nitrit, oxydase, catalase va cae thude thir khac duge sir dung dé ching minh cdc dc tinh sinh héa phai dug kiém tra xac nhan higu nding truéc khi str dung. Cac chiing 4m tinh, dong tinh phai duge sir dung 48 éanh gid xée nhan va han st dyng phai duge thiét lap. Can sir dung céc thudc thi thudc loai tinh khiét phan tich cho caTCVN 8128:2015 phép thir khang dinh. Néu sir dung cc loai thuéc thir thong mai, thi tuan thii céc huréng d&in cia nha san xuét va bo quan va siz dung [18], [19]. 7 Phuong phdp thir higu nang cla méi trrdng nudi cdy dic 74 Yéucdu chung ‘idu nay m6 td phép thiy higu nding dinh tinh va dinh long ai voi mdi trux’mg nubi ody de urge quy inn trong cdc tigu chudn cy thé dp dung cho thye phdm va nude. Day la cde phyong phap chung déi voi hau hét cdc méi truéng nudi cy. Cac phuong phdp nay cé thé khéng thich hep cho phép thir mot 86 kidu mdi trudng vé do thu hdi nfm méc. So ad tom tét ca phurong phap 6 trong Phy lye C, 7.2 Phweng phap thir dinh lvgng 7.24 Phuong phép thir djnh lryng — Cac djnh nghia 7.244 Higu sudt Higu suat phai dat dure giéi han x4c dinh t6i thiéu (xem tigu chudn cy thé lién quan hoge 4c Phy Ic E va Phy luc F). Xem Phu luc G vé sir dung so dé kiém soat 48 thir hiéu nang cia mi trurémg nudi cdy dc bang quy trink quy dinh dudi day. ‘D6i voi cdc phurong phép dinh long, hé sé higu sudt P_[21] dug xéc dinh theo Céng thie (1): oM w Trong 8 NN, 1a s6 dém khudn Igc ting s6 thu durge trén hoge trong mai trvéng nudi cdy can thi, nghia la sé ‘4ém khudn lac trén cdc Ba; No la s6 6m khudn lac ting sé thu dug trén méi truéng nuéi cy déi chtrng thu dugc tir mot hose nhiéu dia va phai 2 100 cfu, xem 5.4.2.5.1. Dién gidi cdc két qua, xem 7.2.2.1.2. 7.2412 Tinh chon ioc Méi tng nudi cdy chon Igc va méi trudng déi ching khéng chon loc duge cay cling cdc vi sinh vat khong phai dich & cc d pha loang khée nhau.TCVN 8128:2015 H@ sé chon loc, S 4, duge tinh bling Céng thire (2): Sp=Do- Ds @ Trong 66 Do 18.46 pha long cao nhat cé vi sinh vat phdt trién trén méi trurémg d6i chirng khong chon loc; Ds 18 46 pha long cao nhat cé vi sinh vat phat tin tat trén mdi truréng thoy nghiém chon loc; Sr, Dy va D, duge bibu thi bang don vi logro. CHU THICH: VI dy, néu Do10™ = logios,0 va Ds 10 = logre3.0 thi hg sé chon loc Sr = 1,0. Dién giai két qua, xem 7.2.2.1.2. 7.2.2. Phwong phap thi? dinh lugng dét véi méi trxéng mudi cdy dic 7.2.24 Yeu cdu chung ‘Quy trinh nay edn siv dyng huyén phir vi knudn 43 duce dinh long (66 thé 18 huyén phd mau tho/vat ligu chuan éinh lung duc) cé ndng 46 thich hep voi chiing dich. 86 thu hdi cia mé méi tru’ng nudi cdy mai duge so snh voi d6 thu hdi cia mdi trréng nudi ey khong chon loc (mai trong adi ching) hoc trong cdc trvdng hep dac biét, dugc so santrvéi mé di duge ch4p nhan trudc 46 cla thanh phan moi tring turong ty 7.2.2.4.4 Cach tién hanh a) Str dung céc gidng lam viée va dich cy e4 ning 6 thich hop biét trurée ciia chiing dich va ching ‘cdy khéng phai dich trong 5.3.2 hoe RM thich hop. b) Can siv dung mét ho&c nhiéu dia cho méi vi sinh vat. Sé 6ém duge sir dung phy thugc vao c& cla mé, 46 tin c&y trong quy trinh s2n xudtwtam-bdo chét trong-46 tin cay va mic vi sinh vat trong huyén phi mau thi. ¢) Bam bao rang bé mat cdc dia 1a kn6 hoan toan, xem 4.5.5, d) Nudicdy bang ech dan dich cy trén méi trdng hode baing phurong php loc mang dé cho cac sb dém nam trong cac gidi han khuyén c4o néu trong 5.4.2.5.1 46i véi phép thir dinh Iuong. Phuong phép nhé giot bé mat Miles-Misra cai bién, céc hé théng nhé giot khdc ho’ie d6 dia xoan cling 66 thé duge sir dung 48 66 durgc cac khudin lac 06 thé dém duce trén dia. ~ B61 voi cdc mai trréng nusi cdy thudng dug sir dyng 8 dinh lugng theo c&ch nay thi phai sir dung phuong phap dé dia. — Nudi cdy mdi tréng di chi¢g hoe cdc dia tir c&c mé duge chdp nhan ruse dé theo cdch twong ty. 36TCVN 8128:2015 — U 4m cde ia véi cdc aiéu kién quy dinh trong cdc tigu chudn cy tha. = Bhim cdc khudn lac 06 mat trén méi dia, Danh gid kich thuée va hinh thai oda cdc khudn lac trén ho&c trong mai trizéng thir nghiém bang cach so sénh véi a} thu hdi tran méi trang nudi ody khong cchgn Igc (mi truréng a6i chieng) ho&c m8 44 dugc chdp nhan trrde dé ca thanh phan méi trong tuong ty. 7.2.24.2 Tinh két qua va dién gial két qua Oi voi phép thir dinh lugng, khodng 100 cfu 1a can thiét dé dam bao 4 chym (xem Bang 1), C6 thé can phai si? dung nhiéu dia cho méi lan lap fai, Cac két qua duge chép nhan cé higu lye néu dp teng cdc didu kign sau day: = MBi lap lai phi cho két qué durong tinh (co phat trién vi khudn dich); - MBi két qua b4o c4o riéng 18 bao gdm trong dai phan tich chudn (an 100 khudn lac adi voi cdc phyong phap loc va dén 150 khudn lac di voi cac phuong phap bé mat). a) Pp phi 2 0,50 48 so sanh méi trong chon loc v6i méi truéng 661 chimg khéng chon lec quy dinh trong Phy yc E va Phy luc F, Pp phai 2 0,70 dé so sénh méi trrdng khéng chon Igc voi méi trréng a6 chimg knéng chon loc hoe nh quy dinh trong Phu Wye E va Phy Iyc F. Bidu nay o6 thé 4p dung cho ac ttudng hop dic biét khi so sénh voi mé trade 46, b)Néu Pe vuot qué 1,4 thi phai tim ra nguyén nhan. ¢) Spcda cdc vi sinh vat khéng phai dich It nhdt phai bang 2. Dbl voi cdc trréng hyp dc biét, xem céc Phu luc S, F va |. Cac tiéu chi nay 06 thé khéng thich hop cho dc mi trréng khong dug quy dinh trong cc Phy yc E va F vi dy nhu céc tiéu chi nay quy dinh trong cac tiéu chudn cu thé, 7.2.22 Sip dung hé sé thu héi tir chét chudn Nguyén tac nay sir dyng cdc chat chudn RM, CRM hodc céc CRM néi b6 dé cung ep huyén phi vi Khudn cé chtra mét lugng don vi hinh thanh khudn lac dich 4 biét ho&c céc ching khéng mong muén. He sé thu hbi tty mé mdi cia mdi trréng nudi cdy 66 thé dure so sdnh vai sé Ireng cfu dy kién ty RM, GRM hoac RM ni bé. C6 thé stv dung chénh léch toi han 8 tinh giéi han dung sai (xem TCVN 6910-6 (ISO 6725-6) (34). Xem Bang 1 37TCVN 8128:2015 ‘Déi voi viée chudn bi va danh gid cdc RM néi bé, xem [21] va [29]. Chat Iugng etia RM phai dug kiém tra x4c nh&n trén méi trudng déi chimg, 7.3. Thir nghiém méi trong nudi cdy stv dung cho loc mang Chat luong ciia cdc bé loc mang duce sir dung phai durgc danh gid truréc dé 48 chtmng minh tinh phis gp cila ching vé vige sir dyng, xem ISO 7704, é thir higu n&ng clia mdi truréng nudi cdy str dung trong loc mang, sir dung giéng lam vige va dich cay trong 6.4.2, Nudi cAy méi tréng huy€n phi, vi dy: chat ldng pha loang, née v6 tring véi mie dich cdy thich hop néu trong 5.4.2.5. Loc chat léng theo cac yéu cu cia tiéu chuan cy thé. Dat mang loc lén ba mat thach cn thir. Nudi cy mét Iveng dil céc mang/dia 48 thu duge khoang 100 cfu cho thir nghiém higu suat. Lap lai voi mang loc méi va dat mang tht hai {én b& mat méi truéng déi ching, st dyng cd¢ dé pha long néu can cho phép thir tinh chon Igc. Nudi 4m céc dia theo tiéu chun cu thé. Lap Iai quy trinh méi lan khi thay 661 mé mang oc méi cing nhu mé méi trvéng mdi. Néu cn, anh gid anh huéng ctia mang loc dén két qua sao cho dan éau dich ey thir nghiém lén moi trudéng thir nghiém va mai trréng déi ching ma khong cd mang I9c. 7.4 Phwong phép thir dinh tinh 7.4.1 Phuong php cdy vach djnh tinh 7.4.1.4 Cach tién hanh ‘Sir dung cdc giéng lam viée va dich cy trong 5.4.2. ‘B6i voi cdc phép thir higu sudt va tinh dac higu, sir dung dia méi trudng thir nghiém va cdy vach tung ‘vi sinh vat thi nghigm sao cho thu duge cac khudn lac moc riéng ré. Béi voi cdc phép thir tinh chon I9c, stv dung da méi truéng thir nghiém va cdy vach ting vi sinh vat thir nghiém thanh ting vach thang sir dung vong cy 1 pl lan b& mat mdi trudng thir nghiém. Vai vi sinh vat thir nghigm c6 thé duoc cy vach trén cing mét da thanh céc duréng song song khéng giao nhaw, c&c vach can phan biét ré dé cho phép quan sét hinh thai dién hinh. Co thé str dung cdc phuong phap edy vach @& chuan hoa khac, U 4m cdc dia & cae diéu kién xde dinh trong cdc tiéu chudn cy thé. 38TCVN 8128:2015 TA1.2 Dién giai cdc két qua ‘Banh gid sy phat trién trén cdc dia thach sau khi G nh sau: = Otuong dng voi khong phat trién; — 1 tuong tng véi phat trién yéu; — 2 twong Ung voi phat tridn tét; Céc vi sinh vat dich phai bling 2 va c6 vé bé ngoai, kich thuée va hinh thal cla khuan tac dién hinh. ‘Déi véi cdc phép thé? tinh chon loc, mic 48 We ché phy thude vao kidu moi trréng. Sy phat trién cba c&e vi sinh vat khéng phai dich phai bj tre ché toan phan hoge mét pnan. 7.4.2. Xée dinh tinh afc higu Binh nghfa vé tinh dc higu duge néu trong 3.2.6. Tinh dae nigerede mei trong audi ody 1a chi thi ve cdc Bic tinh vat Iy 48 phan biét cdc vi sinh vat lién quan dén sy od mat, khéng cb mat vashoge cp a> thé hign cdc phan tng sinh héa va kich thuée khudn lac va hinh thai khudn lac. B6i voi cdc yéu cdu cilia giéng lam vic va dich cdy, xem 5.4.2. 7.4.3 Cac phyong phép djnh tinh khac déi vei mél trrng aac C6 thé si? dung cde phuong phap dinh tinh khdc [9], [21]. 8 Phwong phap thir higu nang cia mai trong nudi cdy ting 8.1 Yéu cau chung Didu nay mé td cdc phuong php dinh tinh va dinh lweng dé thir hiéu ning cla méi trréng-nudi c4y Jang. So 4d tém tat cho ting phuong phdp dirgc.néu trong Phutuc.C. 8.2. Phuong phap djnh lrgng trong éng nghigm dé thir higu nang cia méi tnrdng ting sinh léng (phwong phap pha loang dé phan bigt) 8.2.1 Yéu cdu chung Phuong php nay la phurong php chung cé thé duvoc sir dung 46i voi méi truéng léng chon i9c ho&e khéng chon loc. Phuong phap nay cling cé thé thich hop 66 thir hiéu-nding cla méi.triréng léng ding 48 dinh lweng, vi du: trong cac phuong phap dém sé c6 xac sudt lon nhat. 8.2.2. Chudn bj day pha lodng = Chon sé éng dai dién, xem 6.3.1. 39TCVN 8128:2015 — Chuan bj cdc day pha loang thich hgp tir giéng lam viéc cia vi sinh vat dich hoe khéng phai dich trong cc dich pha long quy dinh trong TCVN 6507-1 (ISO 6887-1) va TCVN 9716 (ISO 8199) sao cho Kh6ng cé mat cdc vi sinh vat trong dung dich pha lofing nhét, vi du: tir 10 dén 107°, Thuong si dung ‘4c day pha long thap phan, nhung cdc &@ pha long 1/5 hoc 1/2 cling thich hop. = Sd’ dung cdc day pha long trong khodng thé’ gian quy dinh, xem 5.4.2.3, = Tai thdi diém sir dung, chuyén mét long a biét, vi dy: 0,1 ml mdi dung dich pha long sang bé mat cila dla thach mai rvdng khdng chon Igc va dan déu - U 4m trong cde didu kign thich hop déi v6i vi sinh vat lién quan. — Bém sé khudn lac trén cc dia thach & néng dé pha loang thdp nhat o6 chiva dén 150 khudn lac va sé khuan lac c6 46 pha long cao hon 4@ pha loaing nay va ghi lai két qua. 8.2.3 Quy trinh thir nghiém mél tredng long — Chen sé iwang éng méi trong cn thir twong ting véi s6 éng trong day pha lodng. ~ Sir dung cae dung dich pha long 4a chudn bj theo 8.2.2 va bat dau v6i dung dich pha long nhét, nubi cAy mt long 8 biét ciia huy4n phi vi sinh vat thir nghiém, vi dy: 0,1 ml vao éng méi trxdng trang ing. = U dm cdc éng thir nghiém trong cac didu kign quy éinh trong tiéu chudn lién quan, xem 5.4.2.6. — Sau khi 4m, sir dyng vang ody riéng ré 10 pl cho mdi dng mei trudng 4a udm a8 cdy chuyén sang mi truéng thach khéng chon Igc. ~ U Am cde dia 43 cdy 6 cdc éidu kign thich hop a6i véi vi sinh vat. ~ Sau khi d 4m, kiém tra tirng dia vé sy phat trién hoc khéng phat trién cia vi sinh vat. CHU THICH: B6i vdi vi sinh vat dich, thung sit dung céc dung dich pha loling tir 10% an 10°. BSi voi cdc vi sinh vat khéng phai dich, thing sir dung cdc dung dich pha lo&ng tir 10" dén 10. 8.2.4 Tinh va dién giai két qua Higu sudt ctia méi trréng tang sinh lang chon loc durec dap ting néu o6 phat trién tot (it nhat 10 cfu tie vong Ay dy 10 1) cia vi sinh vat dich thu duge tir dung dich pha long tao ra it hon 100 cfu (trong 0,1 mi) trén Sia ‘4i v6i mai trvéng ldng chon Ic, xe dinh hé s6 chon loc Sr, tir dung dich pha loaing nhét ctia giéng lam viéc cho thay phat trién tét (it nh&t 10 cfu) trén dia thach va dung dich pha long nhat cia méi truréng Idng chon loc cho th4y khong phat trién (hodc it hon 10 cfu) clia vi sinh vat khéng phdi dich trén Gia thach khéng chon igc. Hé sé Sr it nhdt phai bang 2. CHU THICH: Cée phone phip bé sung cho phép thir cinh hucng cia mi trubng l8ng ding 48 Binh gid méi trréng dang ‘fuge nghién cit hose trong ec nghin cd so sdnh due nu trong Phy ue! 40TOVN 8128:2015 8.3 Phuong phép djnh tinh trong dng nghigm dé thir higu ning cia mdi trréng long chon loc 8.3.4 Yéucdu chung Phuong phap nay sir dung vi sinh vat dich, khong phai dich hogc hn hop vi sinh vat dich va khong hai dich trong cling éng nghi¢m. 8.3.2 Cach tién hanh = Chon mét long cac éng, mbi éng chiva 10 mi mai trvéing ho&c cdc phan 10 ml cia mdi mé can thir nghigm (xem 3.1.2 va 6.3.1), Tién hanh tiép theo mé td sau day theo cdc yeu cau quy dinh trong cc Phy luc E va Phy lye F. —- Chudin bj dich cy: xem 5.4.2.3, ~ Nudicdy-cac vi sinh vat dich: Nudi c4y mét éng canh thang thir nghiém bing cach cho mét lveng dich c4y chira < 100 cfu vi sinh vat dich réi tron. - Nudi cdy c&c vi sinh vat khéng phai dich: Nuéi cdy mét dng canh thang thir nghiém cla mét vi sinh vat bang cach cho mot lugng dich cdy chira lugng vi sinh vat cao hon (> 1000 cfu) rai trén. — Nudi edy cde vi-sinh vét dich va khéng phai dich trong ciing éng nghigm khi yéu cu trong Phy luc Eva Phy Iyc F ho&c khi dénh gid moi truréng méi ho&c dénh gia nha san xudt méi. Nudi cdy mot éng canh thang chira < 100 t8 bao vi sinh vat dich va mét lung Ién hon céc vi sinh vat khong phai dich (> 1 000 té bao cho mi éng) trong cing éng nghiém va tron. — 04m cde dng nghiém & cdc didu kién quy dinh trong tiéu chudn cy thé, xem 5.4.2.6. — LAy mét vang oy (10 pi) tir dng nghiém chia vi sinh vat dich va c4y vach Ién dia méi truéng khong chon Igc (vi du: TSA). — Néu sir dung hén hop méi treéng nudi cAy vi sinh vat dich va khong phdi dich thi lay mét ving cdy (10 pl) va.cdy vach lan dia mdi trading xée dinh d6i véi vi sinh vat dich. — Lay m@t vang cy (10 ju) tte gidng vi sinh vat khOng phai dich va cy vach én dfa mdi truréng chon loc (vi dy: XLD). ~ U dm cdc fa 64 cdy & cdc didu kién quy dinh trong tiéu chuan cy thé. Néu st? dung thé tich mai trréng lén hon thi nguai sir dung 6 thé chon cach 8 chinh ty Ié dich cdy ‘sa0 cho thu duge céc két qua trong dong. atTCVN 8128:2015 8.3.3 Tinh va dién giai két qua Higu sudt clla canh thang thir nghiém léng la dép dng néu o6 phat trién t6t (it nhat 10 cfu hodc dudng cy phat trién tét) cia vi sinh vat dich thu duge tran méi trurding dae tha déi véi vi sinh vat 46. Tinh chon Ipc cuia canh thang thir nghiém duoc dap ting néu khéng cé siz phat trién (hodc it hon 10 cfu) ca vi sinh vat khong phai trén mdi truéng thach khéng chon loc. 8.4 Phuong phap éinh tinh trong éng nghiém don (49 dyc) 68 thir higu nang cia méi trréng }éng 8.4.4 Yéu cdu chung Phuong phép nay thich hep dé thir higu ning cla méi tnaéng mudi cdy Ing khdng chon loc va méi truzng chon loc 6 thir khang dinh, vi dy canh thang lactose mat luc sng (BGBLB) [41]. Phuong phap nay chi dinh tinh va két qua chi 1a chi thi Méi trurémg duc chi c6 thé duge thir nghiém bang phuong phap nay néu dug cdy chuyén sang méi trrdng dic cho thdy co phat ‘Béi vOi cdc mdi trudng trong, dénh gid nhur sau: = Otuong ting vai khong dye; = 1 tuong ing voi hoi duc; = 2twong ting voi khé duc; 8.4.2 Cach tién hanh 8.4.2.4 Méi trong tang sinh so bd = Chon mét 86 dng, méi éng chiza 10 mi méi trong hode cdc phan 10 mi cia méi mé can thir nghiém (xem 3.1.2va6.3.1). ~ B6i véi phép thir higu n&ng cia mal trréng tng sinh sorb6; vi dy-muée dém pepton (BPW), nudi cAy méi truéng véi mét lung dich cAy thich hep (xem tigu chuan cy thé) ¢6 chira s 100 cfu trye tiép vao méi truténg ther nghigm. — Chuan bj dich cy: xem 5.4.2.3, — Am cae éng trong cée didu kién quy dinh trong tiéu chudn cu thé, xem 5.4.2.6. — Kiém tra mdi treéng vé sy phat trién, 8.4.2.2 Méi trréng khang djnh — Béi véi phép thir higu nang cia méi tru’ng khang dinh dang King, nudi cdy méi trrdng thir nghiém voi huyén phu giéng lam vide (c6 chera > 10° cfu/ml) sir dung vong cy 1 pl. 42TCVN 8128:2015 — U dm éng nghiém & cdc didu kign quy dinh trong tiéu chudn cu thé, xem 5.4.2.6, = Néu m6itruéng chura nudi cy bj duc thi c4y chuyén sang mdi truéng dc, G dm cde dia & cdc didu kién quy dinh trong cdc tiéu chudn cy thé va kiém tra sy phat trién, 8.4.3 Tinh va dién gial két qua Thy hign anh gia dinh tinh tryc quan bang cdch quan sét 46 dyc (nghfa la 2) cho thay phat tridn tt, xem 8.4.1, Banh gia dinh tinh méi truréng me dyc ld chT thj cb sy phat trién tren mdi trrdng dac. CHU THICH 1: 6% khi ay phét tin cla vi sinh vat ob thé quan s&t duge khi cb ngung kéviéng & day éng nghiém hoe chai. ‘Trong wong hgp nay, [de c&n thi thi 6 th8 cai thign vige dnh gia va gi thich Kt qua. CHO THICH 2: Céc dic tinh Khao, nbur sith Kh va @8i mau etng c6 thd dah gid bling phurong php nay. 9 Phuong phdp thir higu nang cua chat pha loang va méi trvéng van chuyén 9.1 ‘Yéu cu chung Céc phép thi higu nang vi sinh vat phai duoc thu hign trén mu dal dign cho mé san phdm cudi cing. xem 6.3.1. 9.2 Phuong phap thir nghiém dich pha long 9.2.1 Phuong phap thi? dinh lugng dich pha loang 9.2.4.1 Yéu cdu chung Phuong php xac dinh higu nang cia dich pha long 48 duy tri sy s6ng etia oéc vi sinh vat ma khong 96 sy tang ho&e gid qué mirc vi sinh vat trong sudt qué trinh tiép xic truée khi 43 thach [én dla hodc nudi cdy trong méi tréng léng. 9.2.4.2 Gach tién hanh Nuéi cy phan thir nghiém (vi du 9 ml) dich pha long véi 1 mI huy&n phil vi sinh vat chira khong 10* cfu/ml va tron, 48 chudn’bi dich cdy, xem 5.4.2. Lay ngay 0,1 mi dich da cdy va dan déu trén bé mat thach khong chon Igc (mdi tring a6i ching) nh TSA (t). Gir dich nudi cdy & nhigt d@ phong trong Khong thoi gian quy éinh trong phan thich hop cia bo TCVN 6507 (ISO 6887) hoc TCVN 9716 (ISO 8199) tinh tir théi diém két thc chudn bj huyén phi ban dau dén khi lay dich cy ra cho tiép xiic véi mai tru’ng nubi cdy (thudng 14 45 min). Tron va lay ra mét long tong ty (0,1 ml) va cdy dia Iai trén mdi tréng 441 chiang (4). U dm méi trudng a6i ching & nhigt 46 va thai gian thich hgp, vi du: 30 °C/72 h. 43TCVN 8128:2015 9.2.1.3 Doc va dién gidi két qua ‘Sau khi U 4m, 44m cdc khudn lac trén céc dia f va b. ‘$6 lugng vi sinh vat, fy, sau khi G Am dich pha loang 44 cAy phai trong pham viz 30 % s6 aém ban dau (ts). 9.3 Prong phap thir noi trong van chuyén 9.3.4 Yeu cdu chung Phuong phap xde dink higu nang cia méi truéng van chuy&n a8 duy tri sy séng cla cdc vi sinh vat trong qué tinh van chuyén ma khéng fam ting hod gidm mirc vi sinh vat da cy. Néu cdc ha théng van chuyén phi hyp véi thidt bi |4y mau, thi phai duge sir dung dé cdy méi trudng van.chuyén.-Mat khdc, vide nudi cy phai dug thy hign trong cdc didu kién tuong tng véi cdc dibu ign thyre té U dm méi trréng van chuyén a8 cdy & nhiét 6 va thai gian thich hgp theo thye té hodc néu quy dinh trong fiéu chudin oy thé [12], (13}. VIDU: Gaene théng van chuyén 48 van chuyén mau trong cic diéu kign lanh duge thiy nghlém & mhigt a 5 °C + 3.°C abi \v6i qua trinh van chuyén thong theéng, vi dy: 24h trvéc khi lp ka a8 da 9.3.2. Phuong phap thir dinh lugng mdi trong vgn chuyén long 9.3.2.1 Cach tién hanh Nuai cy phan mu thir (vi du 10 ml) m6i trvdng van chuyén Ing vei mét Ivgng vi sinh vat thir nghi¢m thich hop cn str dung, Str dung mirc dich cdy chiva khang 10° dén 10° 8 bao cha méi éng 10 mi, 48 chudn.bidich-cdy, xem 5.4.2. Lay ngay 0,1 ml mdi trrémg d& cy va dan déu trén b& mat thach khong chon loc (mai trxéng déi chieng) nhu TSA (). Dé nubi cdy trong cdc hé théng van chuyén bing cdc dung oy lay mu, dat dung cy ly mau trong mot Iurong dung dich giéng tam viéc thich hop (vi dy 0,1 ml déi véi tm béng) (chtra khoding 10° dén 10° 18 bo) trong khoang 10 s, sau d6 nudi cdy méi truéng van chuyén bang dung oy dy mau, Lay ngay 0,1 mi mai trréng van chuyén léng 6a cy va dan déu trén bé mat thach khéng chon loc (méi truéng déi cheng) nhu TSA (t). U 4m moi trréng van chuyén 8 cAy & nhiét 46 va thoi gian thich hop theo thyc 8 hoc néu trong tidu chuan cy th, vi du: 25 °C/5 ngay déi voi méi truéng van chuyan va 30 °C/3 ngay a4) véi mbi trang d6i cheng, sau d6 lap lai viée 63 dia trén m6i trvéng déi ching (t,).TCVN 8128:2015 9.3.2.2 Doc va dién giai két qua ‘Sau khi & 4m, dém cdc khudn lac trén cdc dia fy va h. 86 lugng vi sinh vat, f,, sau khi U 4m mdi trveng van chuy&n phai trong pham vi + 30 % sé dém ban dau (4). 9.3.3 Phuong phép thir dinh tinh mai truvéng van chuyén ran 9.3.3.1 Cach tién hanh Nui cy phan mau thir (vi dy 10 ml) mai truéng van chuyén rn voi mét krgng vi sinh vat thir nghiém thich hop edn str dung. St dung mire dich ey chira khoaing 10* dén 10°, 48 chudn bj dich cdy, xem 5.4.2 Dé nudi cdy cdc hé théng van chuyén bang c&¢ dung cy lay mau, dat dung oy ly mau trong mét Iveng dung dich giéng lam viéc thich hep (vi dy 100 pl déi véi tm béng) (chika khoang 10* 6&n 10° té bdo) trong khong 10 s, sau d6 nu6i cy mdi truéng van chuyén bang dung oy lay mau, U 4m moi trong van chuyén 44 nudi cay & nhiét 46 va thai gian thich hgp theo thire té hode néu trong tigu chudn quy dinh ey thé. Céy chuyan lén mdi truéng thach khéng chon Ic (méi trrémg ai chimg), nhu thach TSA va d 4m theo thyc té hoae néu trong tiéu chun cy thé, 9.3.3.2 Doc va glai thich két qua ‘Sau khi d 4m, kiém tra sw phat trin cia khudn lac trén thach khéng chon Igc. Phai quan sat thy sip phat trién cia cdc vi sinh vat sau khi U dm. 10 H@ thdng tai ligu va cdc két qua thir nghiém 10.1 Théng tin tie nha san xudt Nha san xuat ho&c nha cung cdp méi trveng nudi cdy phai cung cdp, khi co yéu cau, ve céc dc tinh phat trién lia vi sinh vat cu thé va cac théng tin chung lién quan dén mé mai trong nudi cAy cu thé. 10.2. Truy xudt ngudn géc Tat c& cdc sé ligu tir phép thir higu nang théng thudng can dug lap van ban theo cach thich hyp va “Gu¢e git trong khodng thai gian thich hyp theo hé théng chat lugng duge sir dung. Can-sirdyng cdc biéu mu kiém soat d6i véi tai liéu va bidu mu 48 @énh gid cdc két qué thi nghiém (xem Phy luc D). 45TCVN 8128:2015 Phu luc A (Tham khdo) Tén goi cdc cdc thanh phan moi truéng nui cdy sir dyng trong phan tich vi sinh vat thye phdm, thire an chin nudi va nue A‘ Yéu cau chung Céc tén goi sau day 64 dug théng nhdt 48 hai hda tén goi cla cdc thanh phan khde nhau cla moi truéng nudi cy trong ¢ac tiéu chudn phuong phdp phan tich vi sinh vat. A2 Pepton = San phdm thay phan casein bang enzym; CHU THICH 1: San phdm nay bao gdm cd sn phdm thiy phan casein bing chit phan hiy tuyén tuy hode pepxin, edn phdm ‘thay phan casein va trypton bang trypsin. - San phdm thiy phan dau twong hod bét dau trong bang enzym; = San phdm thiy phan md déng vét bang enzym; CHU THICH 2: San phdm nay bao gdm c8 pepton tht, s8n phdm thy phan thit bing pepxin, san phdm thy phan thit bing ‘chit phn hly tuyén tuy. — San phdm thily phan tim bang enzym; — San phdm thily phan gelatin bing enzym; = San phdm thiy phan md dong vat va thye vat bang enzym; CHU THICH 3: Sn phim nay bao gdm cd typton ~ San pham thy phan casein bang axit. AS Cac chét chiét va chat dé pha — Ch&t chiét thit va canh thang thit; = Chat chiét tim-ndo va canh thang tim-ndo; = Chat chiét nm men, 46TCVN 8128:2015 A4 Thach Thach dung cho vi khuan, AS Cac loai khac = Nhti trong !ong 63 trig; = Sia bot gay: — Huyét, huyét 48 tach fibrin hoe huyét 48 dung gidi, bot mau, plasma, soi huyét, haemin tir cae G6ng vat xée dinh; — Mat bd ding cho vi khuan; = CA mudi mat, CHU THICH: Cac logi nay-bao gém.cic. mudi mat sd 3. 47TCVN 8128:2015 Phy luc B (Quy dinh) Chuan bj géc déi chieng va giéng lam viéc B.A Chuan bj gée déi ching tir ching 461 chieng abi chong nhan dug Kim tra va luu tri tai figu Hoan nguyén theo huéng dan lia nha san xudt® Phin igp vén tach mau cin 5 %, | TSA hoi m6 tutmg rn thich hgp| Kiém tra 66 tinh khidt va cac tinh ‘ch&t dic tung cia ching ® KHONG Pha hop co ¥ ‘Thu sinh knéiva chudn bj huyén ‘his trong mei truémeg thich hop Phan phdi vo cc dng cryo * va ding anh ® i * hin chung, noan nguyén trong eanh theng cinh duréng va chn thoi gian a8 h&i pyc. > idm tra xée nhgn hinh thé cde Khuen le va nhum Gram hoc nin bit sir dung phép thi sinh hoa © Vi du mi trutig bio v8 eryo, nhu TSB due b8 sung ti 10% glyxerol dn 15 % ghncerol (thd ch). (Cac dng exyo 0 thd chia cdc het * Lam aéng lanh & nhigt dp cri - 70 °C 06 thé Kéo dai thoi gian bo quén. Nén han ché thai ian bo quan & mie & cao hom (36) ' Cothé str dung trvc tgp giéng lam viée, Hinh B.1 - So dd chudn bj géc 461 chieng tir chiing 6i chieng 48TCVN 8128:2015 B.2_ Chudn bj giéng lam vige tir ching déi ching Ra Gong nhanh hoae hoan guyén theo huréng d&n cila nha cung cp —— 1 Tay , | Cay len mdi tudng dc thich hop Céy len moi trudng Bac thich hop | Tay chon 1) = Gidng lam vige chon 2° I I Kiém tra 06 tinh Khiét va hinh thai idm tra 06 tinh Khiét va hinh thai (va cée dc trung néu tt» ngudn (va.céc dc tng néu ty nguén thyong mal) } thong mai) mane KHONG co Cy lén mdi trréng thich hop * ogc dng kh huyn phis® Gidng géc Phan ap tren mi trong thich hop Gidng lam vigc Kid tra 46 tinh hit ° Kid tra cdc ta ligu, XB ca lién kBt dfn ching d6i ching va cde dc tinh lién quan, néu gée d6i ching thu durge tir ngubn kha, ° Quy trinh nay ld thich hyp. © uy trinh nay 6 thé cd thi i voi mot vai ching, vi dy: cho ede phép dinh lxgmg. Mol gai doan phai rec ghi * Vi dy: nubi ey tn thach nghiéng TSA hoe TSA huyét clu hog mal truéng thich hop khée, & Am 24 h va bio quan & nhigt a thich hop (18 °C dn 25 °C ho8e2 °C etn 8 °C ty thube vio vi sinh v@l) dn 4 tudn [38]. * Vi dy mei tardmg bao vB cryo, nh TSB due BB sung tl 10% glyxoraldén 16 % glyxerol ind tch). Lam déng lan & nit 5 duti ~ 70 °C cé thé kéo dai théi gian bdo quan. Nén han ché thoi gian bao quan & nhigt do cao hon [36]. Hinh B.2 - So dé chuan bj gidéng lam viée tir géc 441 cheng 49TCVN 8128:2015 Phy luc © (Quy dinh) So 48 cita cde phwong php thir higu nang C1 Yeu cu chung Xem idu 7. €.2 Phuong phap dinh lweng dét v6i mai trvéng age: higu suat va tinh chon loc (xem 7.2.2 va Hinh C.1). = { chmdr'binéng 68 den oy thichhop, xem 5425.1 | | ogc vat feu chun thich hop Cy trai moi trong thir va mol truing 66i ching howe lke mang 48 c6 $6 dém ‘nam trong dai Khuyéin cdo theo §.4.2.6.1 (Px) hoiic 6.4.2.5.2 (Sr) x. 1 vei tho’ gian va nhigt do quy dinh trong figu chun ign quan ‘ “im khudn lac tr6n cd hai mbitruéng va tinh Pp theo 7.2.1.1 ‘vaKnithich hop tinh S; theo 7.2.1.2 CHO DAN Pa He 56 higu sub Sr Hé-86 chon loc. He s6 higu aut Pa 06 thé dupe dng 68 so sinh: 13} mi trutng khéng chon loc voi mbi tnxng Ai ching khong chon lec. bb) m6i treéng chon loc wi moi trerong di chréng khéng chon Ke. 0) méi trurdng chon ioc voi mbi trrdng adi chimg chon loc. 26 d dm (nop tno ba cia Hinn ndy), xem 5.4.26. Hinh C.1 - So dé phép thir dinb hrgng méi treéng nudi cdy dac 50TCVN 8128:2015 ©.3 Phong phap dinh lrgng trong éng nghigm 48 thir higu nang cua mél truéng ting sinh dng (xem 8.2 va Hinh C.2) ‘Chun b| dy dung dich pha lotng thich hop ti giéng lam vige cola vi sinh vat dich va khong phai dich, vi dy tir 10-"aén 107° ¥ (Cy bng mol turéng thir va edy ala mdi trrdng ai ching ‘voi mdi d& pha logng 2 vor thet gian va nhist 60 avy dinh ‘tong iéu chudin fen quan ¥ Ly ra m6t long, vi dy 10 pati m&i dng da 0 ‘va cy thém vao mdi triréng khéng chgn lec a ‘O.voi thot gian va nhiet 69 quy din trong teu chun ién quan + -Bém khudn lec, tinh higu sudt va 4 chon Igo. theo 8.24 CHU THICH: ©4 & 4m (hop thir nam cia hinh nay), xem 6.4.2.6. Hinh C.2- So 4d phép thir higu nng moi tru’ng tng sinh lng (phwong php 46 pha loang giam dan) 61TCVN 8128:2015 C4 Phuong phép djnh tinh trong éng nghiém don déi véi méi truéng ting sinh léng chon loc (véi vi sinh vat dich, khéng phai dich, hogc hén hep cia vi sinh vat dich va khéng phai dich trong cling m6t dng) (xem 8.3 va Hinh C.3) pIcH KHONG PHAIBICH ‘DICH VA KHONG PHAI DICH Ging lam vige ofa vi sinh vat (Ging tam vige cla visinh vat ‘Giéng lam vigc ala vi sinh vat eich dich trong pha tinh ‘e35nq phai dich trong pha tinh valving phi dich tong pha Soh oa Cy mot éng canh thang thir ‘Cy mét bng canh thang thir mot ng canh thang thi a : iighigen v6i's 100 18 bao vi sinh nghiém véis 100 16 bao ighigm véiz 10° th bao vat eich va = 10°18 bao bao vi sinh vat khong pha dich x + | Uvéithdi gian va nhist ap Uvei thor gian va nhigt 60, U voi thoy gian va nhiet 6 ‘quy dinh trong tiéu chuan ‘quy Ginh trong tigu chun quy @inh trong fiéu chun lign quan lien quan ten quan I ] J Ly 10 ulva chy vach vBo da Ly 10 pl va ody vach vao cia Uy 10 pI va cy vach vao da ‘moi truémg khong chon loc ‘mdi truong khéng chon toc moi trating chon foc 46i vot vi sinh vat dich ¥ ¥ ¥ 1U.v6% thar gian va niet ad voi thoy gian va niet 2 U voitho’ gian va nhigt 06 ‘quy dinh trong tigu chun uy dinh trong tdu chuan uy dinh trong tiéu chun Jen quan fién quan lien quan J J ‘Sy phat tién oda vi sinh vat dich ‘Sy phat tin eda visinh vat ‘Su phat tién cia vi sinh vat dich) ‘An dot it nhdt 10 khudn lac khong phai dich cn bj ire che cn dat it nh&t 10 khudin fac va _ hoan toan ho&e mot phan ‘sy phat trién cla vi sinh vat (it nhdt 10 khudin lac) khéng phai dich cn bj (rc ch& hhoan ton ho&e mét psn (itrihdt 10 khudn lac) CHU THICH: 88 0 dm (hop thir ndm cia mdi oft trong hinh nay), xem 5.4.2.6. Hinh C.3— So 43 cia phurong phap dinh tinh trong éng nghigm don 461 v6i mdi trvéng tang sinh lang chon locTCVN 8128:2015 ©.5 Phuong phSp dinh tinh trong éng nghigm don d6i voi méi trxdng khéng chen lec va moi trurémg léng chon Ig: BS duc (xem 8.4 va Hinh C.4) Gibng am vig trong pha tinh J Céy canh thang thir nghiee O41 pl ging | Wve thei glan va niet quy éinh trong teu chudn | en quan | aM Nhin va danh gid 86 dye cia moi trvong mudi cy ling. DSi \v6i moi trong 08 s8n 6 diyc thi cdy them vao moi trong 8c va Kiém tra su phat tin iia khudn tac Nhin chung, cAn kiém tra cc dc tinh phat trién vi khuan 464i véi ting méi truvng léng, vi dy: sinh khi, 441 mau, néu c6 thé, CHU THICH: 88 G 4m (hOp tha ba cia hinh nay), xem 5.4.2.6. Hinh ©.4~ So dé thir dinh tinh mdi trzeng long trong éng nghigm don (49 dyc) 53TCVN 8128:2015 Phy tye D (Tham khdo) Vidy v8 phiéu ghi céc két qua thi nghigm méi trong nudi cay Bang D.1 - Vidy vé phiéu Phidu kiém soat thir nghigm chat lwgng ngi bd cia mdi trvng nudi cdy ‘Moi trudng nudi cdy ‘Luong dugc: Ngay rot ‘$6 md ngi bd chun bi ‘Moi trutng kho (va ma $6): Nguoi cung cdp | Me Lirgng: Ngayicho ky | ‘Chat bd sung Noveiccung ep | ME Luong: Ngayicht ky Chi tit quy tinh: Kiém soat chat lugng vat ly ‘Ve bén ngodi dy Kién cia moi | Mau binh thutng, | Chatlvgng dye | Cac Khuyéttat: | Ngay/cht ky trang nudi cy khd khong bi chay | khing éinh: _ co Khong: Gia ti pH dy Kid: pH do dug: ‘Chat iugng vec | Cacknuyéttat. | Ngay/cht ky hing dint C6: Khong: Livong dign day vashoac 66 | Quan satduge: | Chat luong dre | Cackhuyét tat | Naay/chor ky day die kién knding dink: C6:__Khéng: ‘Mau sic diy kién Quan sat Guge: | Chdt long duoc | Cac khuyét tat | Ngay/chr ky khang gin C6:__Khong: ‘BO nétisy co mat cla cdc vat | Quan sat dug: Chat lugng duge | Cac khuyét tat: Ngay/cht ky phat quang dy kién khang dinh: C6:__Khéng: ‘DO dn dinh geVGp dcidg. 4m | Quan sat duec: | Chat lugng dues | Céckhuyéttat | Nady/cht ky khang dir C6. Khong: Nhiém khudn ‘$6 iugng dia hoac éng duge =| Két qua Chat ugng dug | Sé lurgng dia’hoae | Ngay/cho ky thir nghigm: khang dinh: }ng bj nhi&m: vam C6: Khong: Phat trién vi khudn — Higu sudt Phuong phap kiém chirng: Binh krgng: Binh tinh: ‘Ching Tiéu chi: Két qua: ‘Chit lugng duge | Ngay/chtr ky Ode khang dinh: Mbi trréng ai cheng: co: Khong Phat trién vi khudn ~ Tinh chon loc Phuong phép kidm sat: Binh lugng: Binh tinh Ching Tru chi: Két qua: Chat lung duoc | Ngay/cht ky Uém: khang dinh: Moi trong 66i ching. C6: _ Khong: Phat trién vi khudn — Tinh dc higu Phwong phap kiém chéing: Binh Ivgng:_ Binh tinh: Ching Tieu chi Kétqua: Chat ugng duge | Nagaylcher ky U dm: "khang dinh: Méi trrémg abi ching Co: Khong: Banh gid mé mol trang Chi fidt Bao quan: Bignlaiciamé hang: co:__khong: _| Noay/chor kj 54TCVN 8128:2015 Phy lye E (Quy dinh) Cac vi sinh vat thir nghigm va céc tigu chi higu nang déi voi moi throng nudi cay khudn lac dug sir dung cho vi sinh vat trong thye pham Phy luc nay cung cp théng tin v8 méi trudng nui cdy, céc didu kién nudi edy, cdc vi sinh vat thir ghiém, $6 9 suu tap gidng vi sinh vat thir nghiém va cdc phan ting dy kién khi thye hién thir nghiém mdi truréng nudi cy. ‘Cc ching dc thi: due chon dé thir nghigm nhdm dam bao sy nhdt quan gitra cée phdng thir nghiém va & ching minh sw khéc nhau gitra cdc mai trdng (gitra cdc mé, gira cde nha sén xudl). Cac chiing ny 48 duoc dénh gid Gay aii dé dam bao su phi hgp va théng nhét vé higu nang. Khi cé nhigu hon mot ching duge lit ké cho mi inh vyc thir higu n&ng (higu sudt, tinh chgn lec, tinh 8c higu), thi it nhdt ec chiing duge sir dung phai duoc viét r6 bang chir b. Cée nha cung op moi ‘truréng thurong mai hea phi thuong mai dy kién sir dung cc ching b} sung, vi dy: cde ching néu ‘trong Bang E.1 8 dm bao thém v8 chat luvong cia méi trurtng nudi cy ma ho ap dung. Bang E.1 due thiét Idp o6 tinh dén cdc ching kiém soat dugc sir dung trong Durge dién chau Au (EP) va cdc khuyén nghj d6i véi mdi trurémg nudi ody vi sinh vat trong thyre pham tir Nhém lam viée cia Uy ban quéc t6 v8 Vi sinh vat thye phdm va Vé sinh thyc phim (ICFMH). Céc tiéu chi nay phai durge dura ‘vo trong ede tiéu chun cu thé khi durye xay dyeng va soat xét. Mé mi trréng nudi cy da durge danh 918 xéc nh§n | mé mdi tnrdng dép tng dug higu nang. $6 lvgng ching égc quy dinh trong Bang £.1 la s6 Kegng tir bang ké cita cdc b} dinh danh ching phd bién do Trung tam Dé ligu Thé gidi vé Vi sinh vat (WDCM) cung cép." Bang ké nay ¢6 chia chi tiét cdc ching d6i chting dure thé hign bang -méi sé WDCM-va céc chitiét lién lac cla céc bd suru tap giéng. Tat cd.cac moi trréngtrich din dure quy inh trong cac tiéu chudin quéc t6 (ISO) va tiéu chudn quéc gia (TCVN). Néu gp phai sy bién d6i v8 ching thi nghién ecu anh huréng cla moi tréng nusi ey (vi dy: thu [ay cling méi trréng tir cdc nha san xudt khéc nhau), thu ldy giéng a4i chieng bé sung tir bé su tap giéng géc. Nguoi sir dyng phai phan hdi vé sy bién adi va ching cho WG5 M6j tnrdng nudi céy cla ISOMTC34/SC9.TCVN 8128:2015 Céc chii thich dg si dung trong Bang E.1 nhu sau a Tén goi dy di cla cdc thuat ng mai trréng viét t&t néu trong Bang E.2. b Cac ching duge si* dung it nhat; © Tham khdo dén bang ké ching d6i ching ca s8n irén httz:ftwww.wfee info v8 théng tin s6 lugng ching cia b6 suru tap giéng va cae chi tiét lién lac. d_ Ching duge chon ty do, mat trong cde ching duge sir dung it nhat. @ L: méi trvdng léng; S: méi trvéng Gac; SS: moi trong ban Gc Phat triénido @uc dug phan loai nhu sau: 0 — khéng phat inénvkhong duc; 1 ~ phat trién yéuthai Gyc va 2 - phat trinfcd dyc (xem 7.4.2.1, 8.4.1). g Escherichia colf WDCM 00013 duc néu trong tiéu chudn cu thé. h Escherichia coli WOCM 00013 Id chat san xudt ra B-d-gluconidase manh va WDCM 00202 chat san xuat ra B-d-gluconidase yéu. i Mét sé quy dinh va han ché ctia quéc gia v8 str dung typ huyét thanh khac. Xem cdc yéu cdu hign hanh vé viée chon typ huyét thanh Salmonetta. J Trong trwéng hop sir dung phép tha’ éinh tinh va dinh long déi voi méi truéng thi chi cin cde két qua dinh lvgng (xem Bang E.1}. k Cac chi tiét v8 kiém sodt chat iveng mdi trong MSRV ké cd nbng 6 cudi cling cila dich cdy va cdc tidu chi dugc néu trong TCVN 4829 (180 6579) | Néu méi trréng dinh dudng duge sir dung cho-hai-hoge ba tng dung khac nhau: it nhat 14 thy hin phép tht phat trién Salmonella (néu phéng thir nghiém thirc hign thir vi sinh vat nay). 1m Néu méi trréng BPW dugc sir dung cho hai hoc ba tmg dung khac nhau: it nhét 1a thee hién phép thir phat trién Salmonella (néu phdng thir nghiém thyc hién ther vi sinh vat nay). 1 Chon cde chting theo phuong phap sir dung trong 46 TSA durgc dung lam mai trvéng a6i ching. 56us Bang E.1 — Cac vi sinh vat thir nghigm va céc tiéu chi higu nang cia mai trréng nudi cay thwéng ding trong vi sinh vat thc pham ‘Mol énrtmg chon ige 6b Gb vi sinh vat Meitwing*| Typ |. Welahvat Tie] choe o Ching dBiehong [86 WOGMc] _Wol_[ Phuong | Chudn cir] Glo phin img ae chin | nang hap ‘mg asi | kid chimg | chang ‘Thach Listeria! S Listeria TCVN ‘Hibu Listeria 00021 * Binh theo Ottaviani monoeytognes | 7700-2 | subt smonocytognos 4a wg ‘iuih jac miu xml: va Agost TSA PRzos ace 18509 Listeria o109 Fem ob quing due monocytognes 1/2 Tinh (44240 Escherichia coi } 00012 hoae Jehenioc} (ars 1)°c 00013 echt = | Binh toh | ton phd - Enterococus 00009 hoac o faecalis * 00087 (DO dae Listeria innocua 00017, - - hud lac mau xenh ty nib Pe ey tam ob qulng ve Baird-Parker | S | Staphylococci dong} TCVN Higu | (24% 2)hdén Staphytococus 0034 ‘TSA ‘Binh Pr>0,5 | Khuda lacmau den Tinh eoagulase | 4830-1 | sudt | @o+2) nv ‘aureus 00032 wong ode xém 8 quang 50 arene trong (phan img arn 8888-1) LL ‘rong long 48 trimg) Tin | aszay E cot 00012 rcchd wan . chonkee| 7 4)°%C ots | 7 [PARMA adn BO dte | (@4+2)hdén | Staphyfococus 00189" Khun tae mau den niu | “(age 2ynv | saprophyticus sisi hobo xdm kheng ob GTEC | staphytocoous | 00038 | fn | | phdndmgamtrong epidermidis tong 68 trina BGBLE L Coliform Tews | He call 00012" ve (2) val 4082 ft 0013 ‘kh ; aso 4831)| - Binh tinh trong dng C6 sinh khi va 65 dyc_ (@222)nébn | Ctrebector found | 00008 Durham (4842) = 0 dc eens mies | ©*9°S | Errorocaccus | 90000 inn tn | 9 oh - faecalis” 00087, 7 khong sinh | oa 8718 NADL ‘Sh02:es Bang E.1 (tiép thoo) ‘Pseudomonas spp. ore Ten | Hig Pseudomonas omh 7138 sudt fluorescens ‘TSA t - Par Os Fs so ete Psoudomanas fragt | _001 41 ven, | 19720) Tic Ecol ‘0012 pian dn | Ee _ chgn loc 00013 - phan (0) D618 Nadmmenvandm | TOVN | Hig Saccharomyces | 00086" mde 9275-2 | sudt corevisioe Reudnigctenda (so Wattemia sebi | oore2® Binh tnung theo timg loa oe he et son | yen, | Pez0.6 [dic tag theo ting sngayt *%P0rzlsrestictus| oor8s Tim | gseiyec |_Eutotinm rubrum | _o0te4 ‘chon loc ‘E.coli 90012 hose 000137 hong: = Bint tin = Baoiius subtiés | 0003 rene) phat én ‘subsp. spzizoni RBC Ndmmenvandm [TOWN | Higu ‘Saccharamyoas | 00058" enbe 92751 | sult corevisige soci (SO |Aspergiius brasensis| 00053" | goa | Dith -| pos gclchd 7 loai 218271) soc chiens: |° Gots long Bc tneng theo ting lod @sen'c |Mucorracemesus | 0181 Tinh Ecol (06042 hase choniec| 00013 Jen ton . Baciflus subtiis 00003 pr subsp. spiizenit EC E. coh TCVN Higa 0012" BS dyc (2) 6846 (ISO| sudt | (24+ 2)haén E. col 00013. Ci Binh tinh |va khi trong| C6 sinh khi va c6 dc 7251) (aa + 2) [6g urna) Tinh | @441)°C | Pseudomonas | 00025 ih tin] _Krng _ chon loc: aeruginosa = phat tien IS(TS) Virnudn igo suit | TCVN | Higu TSA hope 7902 ISO] suat ” |méi tong] 16219) 242 9)nabn Jkrena chor], Clostridium 0007" ‘Binh (48.42) he loc khac Pr20,5 | Khudn tac mau den @7£1)°C Ls ns pone 881 voi mol werd ki wiktuan hi ik ~ Tak cheng E.callt Sora | -— ainnimn| = | Khuda tac khéng 08 'e2b8 NADL $102!6s Bang E.1 (tiép theo) ru dan a ome eee) co | exe ae 4831) (@4z2ynadn 00013 = | Binh tinh temo bng Cé-sinh khi va 06 duc 822 | compacer tours! | coos Cutan Tint Entorococcus | 00009 inh tin | chen ioe foccatid! coos7 | phat 2 E.coli Town | Higu , BO die (2)' 00 ceuetso] sult | oxsanadn | cot | OB | Lop ann YE | co kb (48+ 2) bi Ourham: @7e"C Tinh ‘nterooosus [00008 | Lamang | Khong _ chan ioe faecal! 00087 phat tide ‘mCCDA Campyobactor | Town | Higu Campytobactor | 00158 Khun tae mau xm 7718.2 | sult sejun? 0005} Tech | Dinh | oe 20.5 | nhal, hing va dm, di . ' o, een us Compylabacter cof 00004 mee Ichi 06 nh mau kim foal in| Séhaeng moor2net] Tos aa) ee E.cort nies in tnt Staphylococcus in} - jes cose | = foinn tion ‘MRS: ‘Vi khuan sinh anit TCVN Hig Lactobacilus sake: | 00015” tacfic 7906 so] suse , bmn 15214) ‘Lactococcus lactis | 00016" = Binh Khun lac dae tung Pediococcus 00158 | MRS GB) ong | PR2O7 theo limg loi Gees | Peniosaoous sinh gi (304 1)°C = loweatia| ccaten l9¢ a 7 : « 0013 ta nn Oe . . Bacituacarous | ooo | | ‘MYP Bacillus cereus TCVN Hibu | (244 3)hadn 492080] sult | “C4241 | saciscerous | oon | tsa | OM | rao [Muha lgemauning cd 7932) @o=n%c Tinh 00012 hase We chb toan| chentae] aan Ear cots | [DMR dn) - ne pO es peas] Bacitis abtis [00003 | [anntinn| | knudnige mau vang 8712 NADL sLoz:09 Bang E.1 (tiép theo) Taba ‘Subse. spizizei Thang qutng Ka RPFA [Staphylococst dong] TOVN | Hieu | (@ee22)nabn . Khu fe mau don tinn coaguiase | 4830-2 | sult | cape ane | Stephvlococaus | COOBP | sas | Binh | 2.205 |hosexém ed quing mo| aso B74)°C iaiseus: 00032 wong aye 6880-2 7 ? tinh | aae2yer Ecol’ [oooranode| | nnn | eee . jchon Igc| (37+ 1)°C 00013 phan (0) cg @442habn Sepliiosone on169? Khun lac mau den, feat ‘Staphy = Binh tinh - paceman aang epidermidis oat PPA Pseudomonas epp. | TOWN | Haku Psoudomanes | ors anca | suét fluorescens inh TSA 0s asomrs 48420 Pseudomonas lugng ~ “11082 (25+1)°C aeruginosa 0026 Tinh ‘0072 hod rete en tea oe E.col* Os = {Plattner we Tex E cob dvong tinh | TCVN | Higu ‘00124 O-aucwrondase | 7624-1 | subt col? oootat | tsa | 2 | arog | Studminc maa xann (iso 00202" wena. 16849-) |“Tinh | (@raayw | Entoracoccus | 90008 eon an YATCVN |onontc] (44s 1)°C taecats* 0087 Binh tinh |" ptda ©) = Bo die Citrobacter round | 00006" (so ud ae mau trdng ree49.2) | ide Proucomonas | sosog inh nn] =| a TSC (SC) Clostridium TCVN ‘Higu TSA hog] pertingans 4901 | subt m6i (USO 7937)] ‘trong i. ‘khong Clostridium 00007 Binh perfng chon lc e205 | Khudn ieemau den @tsan "gens 00080 | knaca8i | " (87 £1)°C moi hai Ansong ki khi endo hi Tink 00012 howe roche ean chon loc caf 00013 = Phan Gon; phn (0) - Psausomenas | aoo25 aeruginosa ‘8ZL8 NADL ‘sb02:19 Bang E.1 (tiép theo) vrac |S | Entorobaciedacoae | TOWN | Hee 7 oont2" ssta-2 | suit ot 00013 Kkhudn lac mau hing so Salmon: ‘Binh dén 4, 06 hodc khong 21828-2) ceszzyw | typhymaumt’ | cooar | TS* | wong | #295 | co quingkéttiaben B72 1)°C ‘Salmonetia ‘00030 ngoai Entarticist’ . Tinh Enterococcus | 00000 Sinn inn [Deen __|etoa te] tacelst ous phd ©) VRBL s Coliform TCVN: ‘Higu Khudin igc mau 46 tla, 048,80] subt cot 20012" | rsa | FP | zs | edhodctntng ob 4832) oes oe qudng ket toa. Tin | Gornee | Eneroccecue | oala ~ Tejon wn Cechb oi . chonlgc| faocalis? 0087 pin ©) DO dee ‘Pseudomonas z Khun igekreng mau no 02s Binttion| = a eeaneny Mol ervng khong chen Ipc 4 abm vi sinh vit woltnzong | Typ | Viainn vat Tew | Chie u Ching 61 chimg [88 WOCMe Pol irvine] [chudn c] Cac phan img aac chen | Soa . fsa eaca| an PCA s ‘Bém khudin ige | [$0 4833 | Higu Bacifus subtiis ‘oooog” MPCA subt subsp. spizizoni Gaon Ecol ooo | tsa | OFM | por - Qo#1)"C ‘00013 lugng S. aurous 0024 Mol trrémp ting einh chen loe Méitrxéng | Typ ‘Visinh vee Tu | Choe a ‘Chiing él chimg |$S6 WOCMc [Mai Borg | ‘Chudn o&] Cae phan umg dic . chuan | nang adi ching cs ‘rung chimg se Boron | L | Campytobacter | Tovw | Hou | (enw | Gampylobacter | 000166 77151 | sudt | (72 1)°C sau Jejunf! au hose 00005 A taki hadrian dai soe So cat | coorz | > [BiREaH| ein”) eat dng ve dno gee | pata mCCDA {kh cd anh mau kim log wi +Proteus mirabits | 00023 Tinh E. col” ‘oon12 Ucodinan 0013 = __| Binh tn pre ten : ‘8718 NADL ‘S102:zw Bang E.1 (tiép theo) [aron Too ‘Profous mirabilis | 00025 Ts EE Enterobacierocese | TOVN | Higu E. cof ‘001 sste4 | sult 0013 (so + Enterococcus | 00009 hoa 218264 faecal e008? Satmoneta [00081 hode: >to xnuin| fades mau ning ‘Typhimurium’ 00030 - ‘Binh tinh | lac trén ‘ob quing kat tia bn ‘Selmonelta ‘VREG is saan ee goa a Gran’c ‘+ Enterococcus | 00009 hogc faccecal! 0087 Tinh Enterococcus {00009 ho¥e Ueenb een te teccecafs? 90087 sn tan | 088 PRE, = [in tin) eae - TSA Fraser Listeria TovN | Hieu ‘storia ‘ooo2t™ monocytogenes 7700-1 sudt monocytogenes 4b aioe) +E col — | 00012 hode Khudn tac mau xanh hic on const >10 khuan' lam cé quang mau dyc + Enterocoseus ose cen faecal! 0087 on storia core | = | einntinn | Listoia monecytogenes 1/20 ‘hoo +E. col | 00012 hose Suniel 00013 va Agosti (4822) @res%c | *Exterecoscus | 00009 hot faecalis” 00087 Tint Ecol [00012 hwse Vechs enon oe 0013. toan phi - Binh tinh (0) tren - | TSA Enterococcus | 00008 hede 100 faecalis! 087 wud ~ | Bint th | Tan tsa, - 'B2L8 NADL ‘$102:Bang E.1 (tiép theo) Gilt Canton | Stophylocoedl avon TON Hew ‘aureus ‘oo034* Khuain lac de trang >10 kh] tinh coaguiase | 4830.3 | sult _ | theo img moi trvsng sO e4s2nain | — TE cm | 00012 not tectrtn | "teem TON 4830-1 6888-3) (48:2 2)h/ inh tion] 39k | dso 6888-1yabi vei erence aureus (00032, rake, | Baird Parker va TON +6. Col? — | 000%2 Hote Ror, | 4820-21180 6888-2) 0013, bi voi RPFA) Tinh | (@B22)hy Eco” [00012 hobs techs oan chontoc} (37 1)°C ‘0013 ‘Binh tint | phd ©) - teaTSA HallFraser Listeria town | Higu | (2422)n/ |L monocytogenes db] 00021 monocytogenes | 7700-1 | suit | @oxs)%c +E. col — | oo0t2 hae aso 00013) ds 1129041) >10 ku ++ Enterococcus | 00009 hoBe lectrén Hecat S087 . Aaat | khubn lac mau xanh we| 7 monocytogenes | 00108 Binh ih) Lista |e Soden mau due ta theo Ottaviani +6.coi | ovo12 hoe arate va Agostt + Enterococcus | 00009 nose toocats! ‘00087 Tink Ecol (00012 noe (ecb tata chon te ‘0013, inh tinh | phn 0) - trén TSA ++Enlerocogcus | 00008 hode <100 teocais* 00087 ih th | knusn tac - ‘von TSA Tre Yersinia Town | via | aazayns Yersinia 0088? onterocoltica 12700] suit | @sat)%c | entorocottca > 10 khan} Khudn ac da tung 10273) +E, Co’ | 00012 hobe tinn | '#etrén | theo ting moi trong 00013 Binh tint | iN ode | (vem TCVN 8127 ((SO +Pseudomonas | 00025 sa0c: 10278)) ‘soruginosa Tinh Pseudomonas | 00025 eend toa chon oe| aeruginosa inh tinh | phd O) - Proteus mirabtis_ | 00023 ben TSA 8218 NADL ‘sLoz:Bang E.1 (tiép theo) mets] ‘Satmoneta | TOWN | Hu | (axajn/ ‘Saimoneta . 4820180} su&t | a7 s4)°c Enteritidis** 20030 |» 10 khudn| 6579) Saimonote,, | 0003) fectrén | Khubn tac de tung “Typhi tainn umn] LP Rabe | theo img motte +E.co | o0012hoae ‘ '0| rang | [xem TEWN dzy ISO conta chon toc 6579)) +Psoudomones | coo2s its aeruginosa i _ Tinh cof [ovotaneac oehe ang chon i 0013 phan < 100) Bin th adin lac - | ___| tren TA Frierococcus | 00009 hate 10 Khuda faecalis? 00087 Binh tinh | lac én - TSA SRY 3s ‘Saimoneta: TOVN | Hidu | 2x(@24ta)hf ‘Setmonetia ‘00030 Gigt miu 4029080] sult | ais41)%c | Enters" augeies 8579) ving bao veh mau ‘ring xém,| Kha néng cao. khudn inh tinh yc sau 26h] tac ac tung sau kh Satmonelia 00081 ‘cbr an | mudi chy al tren XLD* ‘Typhimurium*! ‘ ving 2 mau dye 38 teairong nto ol Tinh Eco” [oooizhose (cot pa chon ioe ‘0013 ibn vi siot mu Binh toh aoc, - sang of ing bi sue ‘Enterococcus | 00008 hose Knog faocale! 00087 Binh nh paige ~ wws | t | E-corarongtinn | TOWN | Hu | 24aan7 Ecol 0012" . ‘nh ext | Kod lao thay abi B-D-glucuronidess | 7924-3 | sus} 7a ty°c 20013, Biphinh | Sinhaxit | thanh mau vaog 080 Tia Enterococcus | 00008 hose inn uen| Ong . 18648-3) | chon igc! faecalis? 00087 " phat trién “8218 NA’ g10z:$9 Bang E.1 (tip theo) PSE Yersinia TEN | Higa Yersinia ‘00038" enterocolitica 8127 (IS0| sudt enterocolitica 10273) +E Co — | o0012h08e ‘0013 + Pseudomonas | 00025 > 40 knuén| Khudin lac dc tumg senuginosa — — Jejnhtinn| $96t8n_ | theo ting méi tarbng 3 ngay ain 5 Yersinia ‘00160 GIN hodc | xem TCVN 8127 (ISO rgty@4+3)hv | enterocottica ss0c 40273), @541)°C +E.Colf | 00012ho8e 0013, + Pseudomonas | 00025 aeruginosa Tinh Pseudomonas 00025, Ure. ché toén| chon oo} sonuginosa = | Bint tian | prdn 0) = Proteus mirabiis | 00023 enTSA RVS ‘Salmonella Town | Hitu | 2423 ‘Salmonetia ‘00030 4829180] sult | (41,54 1)%C | Entertiio abiasnal 6578) ‘Salmonella | 0031 lactrén | Khudn lec dc tung Typhimariun’ Bint tinh | XLD hod | theo timg mér tredng - 8" | mai trong] [xem TCVN 4829 (150 +E cof ono hota cron oc, ‘6570)) +Psoudomonss | 00025 seruginosa Tinh Ecol | o0oiz hose Ucerbting chon tee 0013 = [en nn i, 10 . teen TSA Enterococcus | 00008 nose |< 10 khudn| faecalis 00087 = [inh tinh | tac tren - TSA SPB Bacitus cereus | TCVN | Higu | essay | Bocifus cereus | 00001 [> tO Khuda] Khudn lec a8e tng rear] | Bate - Jone) Sa | as hog MYP. 21871) Tinh Ecol 00012 hoe |e ch toa! chon ine ‘0013 - pe tan] pn - 8218 NADL ‘g102:99 Bang E.1 (tiép theo) Mi traéng tong Khéng chen lee moi trueng" | Tye"] Visinh vat Tiéu | Chae u Chang déichimg [SS wDcM| Mai | Phuong Che phan ong dae chain | ning teva | PhEP | Cnsin oie trung él kid wan chimg | soit BHI L_ | Staphylococci duran |Tcwn4830-1 Higu | (2442)h/ | Staphylococcus | 00004 inh coagulase jasoesss-t) suat | (37+ 1)°c. aureus —— Jejoninn| 8442 [FCNN 4890-2 aay - es0.6386-2) Bruceta | L | Campylobacter | Tovn | teu | 2ngayains | Campylobacter | 000156 me (S0} suat naiyl | dejan? ogc 00905 Bd due sae cat mbitnine tou] CAmmvobacter cot”) anos | | PinMtinh) “Gay - phan) whi “ Pha ioing voi | L |Ehdtiing pha tong] TCVN | Pha | 45min—1 hy Eco | o00i2hage a mye dich Be seo 180] ane | 20 'C in 0013 fe bigt. vi dy 6887) 25) (> Binh BPWod aes Stapicnceus | ooose | TSA | iy | aco - bromo-cresol- phan) bias so % st oe mn) Ringer daim@t| L | Chdtténg phaleang | TCVN | Pha | 45min—1h7 Ecol? ‘00012 hoge phan tu 6507 180] losing | 20° ¢&n 00013 sa0% ung dich 6887) (at ‘ Pepton ae wee | oe | com = . Musi pepton $word | cx 30% s6| Dung dich aem| ‘adm géc) phosphat Thioglycotat | L ‘Clostridium TevN | Hitu | @teayny Clostridium 00007 Bbaye perfringens aa ('s0| sudt_ | @741)°C ortingens = | Binmtinn | 8 SNe - 7837) sac TSYEB L Usterta Tew | Hieu | @ieayny Listeria ‘aoozt® ‘monocytogenes | 7700-1 | suat | (@5+1)°c | monocytogenes 4b so - Listeria 0109 . 86 due 11200) - ‘ - aoe ‘ronccytogones 2a Binn tion] phan) ‘8218 NADL Sh02:29 Bang E.1 (tiép theo) ‘Mol trrémg phan lap chon loc Mel trading Tye" | __ Vieinh vat Tiu | Chiro 0 ‘Chang dl chang [SS woen"] Moi | Phuong | Chudn cir] Cac phan img de chuin | ning trudma | php trang est | kiém chimg_| chong “Thach Listeria |S “isteria Tow | Higa | caseayns Listeria ‘oozt® theo Ottaviani monocytogenes |7700(80| suét | (721)°C | monocytognes 4b pin tinn| Phat | Kru toc xanh We tam v8 Agost! 11290) Una ‘cis |} = 18 ‘co qulng dye sronocyiognes V2 | Tinh Escherichia co’ | 00012 hofe a eons ene coos |= [initia | toa: pdr - Entorococus — |oo000 hse ‘© toccats’ —__|oo0e7 _| Lo 4 BO abe storia nocua | 00011 ‘Dinh cnn | Phat tn | Khwdn Iec xan yc kam | hiew a nn tinh | tt khong 8 quing aye mCcoal s Campylobacter TCVN | Higu @atayne ‘Campylobacter 00158 >10khudn | Khudn tae mau xm 7716 0 sudt | 41,541)°C Jelunt ‘0008 ~ | Binh tinh} lac tr | nhet, phdng va Am aot ‘ara, al hrina | compyiosecteroot| 004 mCCDA | hi cd anh mau kim lal phir) [Tinh Ecol? [00012 hobe Coat To chon ie| i toan phan | Khudn lc Khong de cots |= inh inh) scam ‘nag phan (0-1), ‘Staphylococcus | 00034 een’ aureus = | Bint tinh | toa pdr A ©) ersmac |S | Eschercnincot | Tov | Hou | @teayns | E.conorsr7 | oot ‘Khun lec trong subt, 0187 7ee6 80] subt | 721°C (chu — [enn nn] Pratt | dang ote mau ndu ving 16654) khSng sinh t@ va dung kinh abe 18) 1mm Tinh ‘Staphylococcus | 00032 hoe Ueché chon io| ‘aureus! 0034 | | pian tinh | to8n phan = © cot 100012 hose | Ue! ad 00013 ting phn | 14gt 56 khudn tac nds = [Penton TO ng phat tin 8218 NADL S102:Bang E.1 (tiép theo) IN Yersinia Town | wigu | @1e3)n7 Yersinia 0038 Khuain lac de tung ssc enterocoftica |8127 ISO] sust_ | (30 1)%c | enterocoftica | go1eo ‘Binh tinn| Phst tiBn | theo ting mi rx 10273) let tin |" 16 2)_| rem TOV 8127 (SO 10273)| Tinh Ecol” [onot2 node Uecnd chen ge] 00013 inn inn | f027 Phan | Khudn lac khéng sec a hoge ting rung phan (0-1) ‘Staphylococcus 00034 Ue one ‘aureus! Dinh tnt | toan phn - © GPC Vibrio spp. ngcaitr | Tovn | Hitu | (@axayns | Vibriovutniieus | o01a7® ‘Khun lee mau ving a Vibrio 7908-2 | suit | (721% inn tin| Phsttién | dure bao quanh do parahwomolyticusy | (ISOS i 16) | nhugm mau vang moi V. cholerae | 21872-2) trutmg Vibrio cholerae non-| 00203 hat wibn | Khun ac 06 tla de ‘t%non-0138 Bin tnn | PA) | B90 quanh do nbuom mau 46 tia mei teva Tinh Ecol 00012 hose Ue chs cian toe} 00018 hod ‘Bin tinh | toan phd - ‘0080 (0) uve! Bacilus cowus | TCVN | Hu | @iza}naén | Bacitus cereus | 00001 inn tinn | Phattein | Khudin ige mau néng od 7903 ISO} sudt_]48 Wy 0+ 1)°C. t6t (2) quang két ida, 2187) Tan Ecol? 90012 node Uc che enone aa ayy 00013 Pen en an grin : @041)%C Bacillus subtilis ‘00003 Binh tinh | Khudn ige mau vang subsp. spizizani = __|rhong of quaing kat tia PEMBA Cactus cereus | TCVN | Fit | @tza)ndén | Bactuscomus | oocoy rs eee 7903 (180) sult | “(aseayny inh tint eat (2) | tam ob quing K&L the 21871) rN Tinh E. col® 90012 hoge Uc ché chon te| 0013 Binh tinh | ten phan a 44h @ Bodie | @721)°C | Bacius subts | o0003 gu subsp. spleteent inhi} =, | Khun lac mau ing ong 6 quan két tia 8ZE8 NADL shor:69 Bang E.1 (tiép theo) S08 Vibrio spp. ngoal ir | TCVN | Higa Vibrio veiniicus | 00187" ~Tonat idn | ,Khudn tac mau xan Vibrio 7905-2 sult Binh tinh | Tbe @ Iycldd tia ob quang két parahaemolyticus’ | (ISO/TS toa V.choterae a1623) Vibrio cholerae non-| 00203" Phat tnién |Khudn lac mau vang 06 Se 3he |_otmenarse Binh ti | 18t@) udng ket ta Tinh Ecol” |oootzhoae een een ee locors Binh tin | toan phan - occao" © Tex E.coiduongiim | TOW | Heo | isan E colt 001 | B-D-glucuronidase | 7024-3 | subt | (4441)°C ooo13" Binh tink Poa na Khun es a anh «so 16649-3) 00202" + Tinh Enterococcus 00009 Phat trién hon le teecai’ —— |o4¢00087 Bin tnt | et) = BO ate Gitrobactortroundii | 00008” Khun ge mau teéng eu Pseudomonas | 00025 Binh tinh] = | Sen anh tyesxanh De vRBG! Enterobacteriacess | TCVN | Higu | (2422) Ecol 00012” seit | mut | aren’ 0013 Khuda ge mau hog 215281 ‘Salmonolia, | 00031 inh tine | PRs tbn | an 8, 5 node Khong ‘Typhimurium’ hose inhtinh | 46 @) | 06 quang két tda ban Salmonella 002 goa Typhimariom! Tink Enterococcus | 00008 o ca ma [ee — [omental xLD ‘Salmonelia Town | Higu | @atayne ‘Salmonella. (00031 Khudn lac 6 tam mau 4820 (1S0] sudt | @741)°c Typhimurium ea den va vidn trong, mau ears) ‘Selmonetla 00090 eee i et my haya Enteritis iu cla moi trong Tink E.col | 00012 hoe Petite ae coors inn ton | Seer, | Khuda emu ang phan (1) Entorococeus | 00008 hoe ecb wan] ‘eocati! 00087 Dinh tinh phn ©) : 87-8 NADL s10z:OL Bang E.1 (tiép theo) Mal trrbng phan ap khong chon Moitvong® [Tye | Vien vet Tieu | Choe 0 ‘Ching déiching |S8WOCM] Mai [Phuong |Chudn eG] Cie phn img dhe ehuin | nang truéng | phép rung él | kim chime | chimg Thech dinn |S | Entemmbacteraceae | TOVN | Higu | (2aeayn/ E ooh ooo dutng . sete aso} sult | @7+4)°%C ‘oon: 21528 (tt phan) ‘Saimonotia TOVN Baa phi ‘Salmonetta ‘00030 _ pina tin | Phatién . 4820 (ISO @724)°C | Typhimurium 00031 164 (2) 8579) Salmonala Entertidis® Yersinia TOVN qe Qne Yorsinta ‘enterocolitica | 8127 (ISO! BIEN S enterocolitica +0273) | TSYEA s Listeria TCVN Higu 14 Hhs Usteria monocyiogenes | 7700 | sult | Grate | monocytogenes 4? eral 180, storia o01ue - Binh tinh | PALE on +1290) monocytogenes 12a sot (2) (Sted cdo phn) Mal trvémg ding cho nhidu mye dich Meitruéng® [Tye] Visinh vat Tidy "| chic u ‘Ching déi ching [séwoow® | mei] Phong Che phan img de chun | ning twang | php fs rng ast | tam |Chudn ctr shimg | _chimg BPW” L_ | Pha loting a8 dém tt) TCVN Pha | 45mindén 1h E. cof 0012" cdvisinh vet 6507 ISO} long | 20°C dén 00013 6287) (at 25°C _ cd che 0034! 20% phan) aureus conn |, tbe TSA laclTa 30) = TCVN Wwond | % sb aém 8507-5 goo) so 6287-5) 828 NADL sL0z:a Bang E.1 (tiép theo) Bw Pha loang a8 afm | TOVN | Pha | (@naS miny/ Utes 00a 200% Listeria 700-2 | toing | (2022)°C | manocytogones inn | Kruée monocytogenes | (ISO ‘asain: coco | 7 | | lectTo - 1290-2) monocytogenes 1i2al PE | (2 30% s6| im ge) TTangsinn sovo8 | Tovn | Higu | (esa | Salmonaia | 0003 Phat rign Salmonetta| 482080] sudt | (@729)%C | Typhimurium! jon tnn| POQHE 8579) ‘Salmonla fy [Pinta af - Enteritidis® Tangsithsobpaé | Town | Ha | (esajw . cof 00012" phattién— | s5iet | sult | ra nec 0013 Enterabocter-acaee| (SO ‘sisisins Po due 21828-1) Typhimurium® | oocst hose | > «| Pinhtinh) “ys - ‘Saimoneita 00032 Enteritidis* Mol trimg dl chimg Gti vai 26 dim vi sinh vit Maluvone [Tye] Visinnvat [| Tieu | hae u ‘Ching G61 chimg [58 WDCM| Md! | Phurong [Chain ca] Cac phan Gg dhe ehudin nang ‘truéeng | php treng ‘abi | kid ching | ching Thachméu |S | Campylobacter | TGVN | teu | (ssa) | Campylobacter | 00166 | Mamoi | inh | Peza7 771s2 | suit | @15a1°C Jejunt coves | tudng | long so Campylobacter cou thech (10272-2) — mau da - duge anh gi i sudt_ | trong phuong bits | trong | ivgng phapmaTsa | crcnissiary | °° | texas ge ding am auge meitraeng asi | Ecol C0082 | gant gis chang €, coli O157:H7 00014 Khudn tge age ees theo img lei sects) usa oot monerytogeres 45 Staphylococcus aureus 00034 ‘8Zb8 NADL S102:ee Bang E.1 (két thc) ‘SDA | 5 | Dbmknudn tae — | Higa | Theoquisinn | Saccharomyces | 000s | Méinai | Binh | F207 ult | cme beg. | teog Khyan tactehdiemam Aspargitus 8c rung theo ting tai dugeding im | jmifonals auge geen i turdng 68 énh gid ching |" Tan goi diy i olka cdc mbi turdng vidt tét néu trong Bang E.2. I Cac chiing duoc thi nghiém it nhdt J Tham kndo bang ké ching Abi ching ob sin tren htp:fnww.wfec.nfo we thdng tn 86 wong chving cia BO suv tp gidng va cfc chi tit en ac: |’ Chiing dure chon 4? do, mét trong cac ching duc sir dung it nhét, J* 1: méi trrémg Iéng; S: mai trudng dao; SS: méi tring ban dc | Phat kidnap ayc dupe phn logl nur sau: 0 —khéng phat biérukhéng duc; 1 phat trién yuo’ duc va 2 phat tidnied dc (xem 7.4.2.1, 4.1) |" Escherichia co¥ WOCM 00013 dugc néu trong tidu chudin cy thd. |" Escherichia coli WOCM 00013 ka chat sén xudt ra (-d-gluconidase manh va WOCM 00202 chat sdn xudt ra B-d-gluconidase ybu Mot s6 quy din va han ché ca que gia v8 st dung typ huyétthanb khée. Xem cdc ytu clu hig hanh vb ige chon typ huyétthanh Salmanot |) Trong trutmg hop si dung phdp thir dinh tinh va dinh Iurgng di v6i mai turtmg thi chi cdn céc két qua dinh long (xem Bang E.1). I Gacchi tbe v8 kim soat chat irom més tong MSRV kd c& ning J uSi cing cba chit cy vd cic tau chi de néu trong TCVN 4829 (ISO 6670). | Néu mol irong din duéng duvge sir dung cho hai hod ba img dung khéc nhau: it nhét ia the hign phép thir phat tién Salmonoiia (ndu phong thir nghigm thc ign tr vi sinh vat nay). |" Neu mei trréng BPW durge sir dung cho hai hogcba tg dyng khéc nha: it nh la thie hign phép thir phdt tién Safmanelin(néu phéng thir nghigm thy ign thir vi sinh vat ny). [P_ Chon eée ching theo phirong phép cb sir dung TSA lam mdi truéng abi ching. SL0Z:8218 NADLTCVN 8128:2015 Bang E.2 — Cac thuat ngir viét tat v8 moi trxémg durgc sir dung trong Bang E.1 ‘Thuét ng mol trvéng Ten day di cila moi trudng Tiéu chudn vidt tit Baird-Parker ‘Thach Baird-Parker TCVN 4830-1 ((S0 6888-1) BGBLB anh thang lactose mat luc séng TCVN 4882 (ISO 4831) BHI ‘Canh thang tim no TCVN 4830-1 (ISO 6888-1) va TCYN 4830-3 (ISO 6888-3) Bolton Ganh thang Botton TOWN 7716-1 (180 10272-1) BPW Nude éém pepton TCVN 6507 (ISO 6887) (tt cd cdc phan), TOVN 4829 (180 8579), TOWN 700-2 (ISO 1290-2), TOWN 5618-4 (180 21528-1) Brucella Canh thang Brucella TOWN 7718 (ISO 19272) {tt od ede phn) cre “Thach xetrimil, fuxidin va xephacioridin TOVN 7138 ((SO 13720) GIN Thach cefsulodin, Irgasan novobiocin TCVN 8127 (ISO 10273) crc Thach xenlulose polymixin B colistin TOWN 7908-2 (ISOITS 21872-2) ” eT-smac Thach MacConkey sorbitol telurt xefixim TCVN 7686 (ISO 16654) bere Thach Dichloran glycerol ‘TOVN 8275-2 (ISO 21827-2) DREC Thach Dichloran-rose bengal chloramphenicol | _ TOVN 8276-1 (ISO 21627-1) EC Can thang EC TOVN 6846 (ISO 7251) EE Can thang glucose mat Iye sng, 06 dm. TOWN 5516-1 (I$0.21528-1) Fraser Canh thang Fraser TCVN 7700-1 (180 11290-1) Hatf-Fraser Canh thang niva Fraser TOWN 8127 (ISO 10273) Is cTs*) Thach sit sulft TCVN 7902 (ISO 15213) (Thach trypiose sult) ie anh thang Irgasan, ticarcilin chlorat TOWN 8127 (ISO 10273) Lst anh thang Lauryl sulfa, triptose lauryl TCVN 4882 (ISO 4831) va TOVN 6846 (ISO 7251) mCDA Thach deoxycholat cefoperazon than cai bién TOWN 7716 (ISO 10272) (tat od cde phan) moro Thach colstin B polymyxin xellobiose cai bién | _TCVN 7905-2 (ISO/TS 21872-2) MKTTA Méi truémg novobiocin tetrathionate Muller- TCVN 4829 (IS0 6578) Kauffmann MMS Moi trréng glutamate khoang cai bién TOWN 7924-3 (180 16849-3) MPCA Trach aém ora cé stra ghythach 6ém dia cb stra|___TCVN 4884 (ISO 4833) MRS Moi trang MRS (de Man, Rogosa va Sharpe) TOVN 7505 (180 15214) 73TCVN 8128:2015 Bang E.2 (két thc) Thuat ngtr méi trvéng Tén day du cia mdi trueng Tiéu chudn vidt tat MSRV Moi trréng rappaport-Vassiliadis ban dae ‘TCVN 4828 (ISO 6579) cai big MyP Thach mannitol long 43 trig polymyxin ‘TCVN 4982 (ISO 7832) PCA Thach ¢ém aia TCVN 4884 (ISO 4833) PEMBA Thach bromothymol mannitol long 48 trang TCVN 7903 (ISO 21871) Pyruvat polymyxin PPA ‘Thach penicillin va pimaricin TOVN 8898 (ISO/TS 11059) PSB Cann thang sorbitol va pepton mudi mat TOVN 8127 (ISO 10273) RPFA Thach fibrinogen huyét tong thd TOVN 4830-2 (ISO 6888-2) RVS ‘Canh thang Rappapon-Vassliadis pepton TCVN 4829 (ISO 6579) au tong SDA ‘Thach Sabouraud dextrose = sps ‘Thach natri dodecyl sulfat polymyxin sucarose | _ TCVN 7905-2 (ISO/TS 21872-2) sspc ‘Thach Salmonella Shigella deoxycholate va TOVN 8127 (ISO 10273) canxi TeX ‘Thach trypton mat X-glucuronit TOVN 7924 (ISO 16649) (t8t 8 céc phan) TCBS Thach sucarase, mu6i mgt, xirat va thiosulfat | _ TCVN 7905-1 {ISO/TS 21872-1) Thioglycoliate Moi trurémg thioglycolet ling TOVN 4991 (ISO 7937) TSA ‘Thach trypton dau tong - TSCISC | Thach sulft xycloserin‘thech sulft xycloserin ‘TCVN 4991 (ISO 7937) | khong chisa long 46 tring TSP ‘Tach polymyxin trypton dau trong TOWN 7203 (ISO 21871) TSYEA Thach chat chidt nm men trypton dau tong TCYN 7700 (ISO 11280) (tat ed cde phen) TSYEB anh thang cht chit ndm men trypton dau TCVN 700 (ISO 11290) twong (tht cd cae phn) VREG ‘Thach glucose mat a8 tim TCVN 5518 (ISO 21828) (tt cd cac phan) VRBL Thach lactose mat ¢8 tim TCVN 6848 (ISO.4832) XLD ‘Thach deoxycholate lysin xylose TOVN 4828 (1S0 6579) 14TCVN 8128:2015, Phy luc F (Quy dinh) Cac vi sinh vat thir nghigm va cdc tiéu chi higu nang déi voi méi trxeng nuéi c4y khuan lac duge sir dung cho vi sinh vat trong nuéc Cac ching dc thi due chon 48 thir nghiém G8 dam bao sy nhat quan gitra cdc phéng thir nghiém va 8 ching minh sy khac nhau gitva cdc méi trvéng (gitra cc mé, gitka oc nha sn xual). Céc ching néu trong Bang F.1 da duge dénh gid day di dé dam bao sy phi hp va théng nhat vé higu nang. Khi cé ahiduchon-mét ching dug ligt ké cho mdi khia canh cla phép thir higu nang (higu sudt, tinh cchon gg, tinh dc higu) thi it nhdt cde chiing dure str dung phai dee viét rd. Cac nha cung cAp méi truéng thong mat hodc phi thong mai dy kién str dyng cae ching bd sung, vi dy: c&¢ chiing néu trong Bang F.1 48 dam bao thém vé chat long cia mai truéng nudi cdy ma ho 4p dung. Céc tiéu chi nay phai bao gdm trong cac tiéu chudin cy thé khi durec xy dyng va soat xét. Mé méi trudng nubi edy 68 duce dénh gid higu Iyc 1A mé méi trwéng dap Ung durge higu nang. $6 lveng ching dug quy dinh trong Bang F.1 [a sé lugng tir bang ké cila cdc bO dinh danh ching phé bién do Trung tam Dir ligu Thé gidi vé Vi sinh vat (WOCM) cung cp. [20] Bang ké nay cé chira chi tiét cac ching déi ching durge thé hign bang mdi s6 WDCM va cae chi tiét lién lac clia cc bd suru tap giéng. Tt o& cdc mdi trréng trich d&n duge quy dinh trong cdc tidy chudn qudc té (ISO) va tiéu chun quée gia (TOVN). Néu gp phai sy bién d8i vé ching, thi nghién iru dnh hugng cia méi tréng nudi cdy (vi dy: thu lay cling méi trréng tir cdc nha san xudt kh4c nhau), thu My giéng d6i chirng bd sung tir bd suru tap giéng g6c. Nguéi sir dung phai_ phan hei vé su bién ai v8 ching cho WGS Méi trudng audi cdy cla ISOMTC3aISC8, 75TCVN 8428:2015 Céc Chi thich duoc sir dung trong Bang F.1 nhu sau: * — Tén goi day di cla cdc thuat ngt mdi truvdng viét t&t néu trong Bang F.2. Cac ching durge sir dung it nhat; © Tham khdo 6én bang ké ching déi ching o6 s&n trén httov/www.wfes.info vé théng tin sé tong chting cla b6 suu tap giéng va cae chi tiét lién lac. 4 Ching dug chon ty do, mét trong cac ching duge sir dung it nhat. * Li méitruéng ling; S: méi trréng dac; SS: moi trudng ban dac 1 Cae chi tiét vé méi truéng Legionella, ké cd bao quan cac ching déi ching, duge néu trong 180 11731. ® Cac chi tiét vd kiém soat chdt Ivong va cdc tiéu chi chdt lvgng méi trréng MUG/EC dugc néu trong ISO 9308, Phy luc E, cén tinh chen loc khang durge quy dinh trong tiéu chun nay. " Céc chi tiét vé kim soat ch4t lugng va cac tiéu chi chat lugng mdi trryng MUD/EC duge néu trong ISO 7899-1:1998, Phy luc E. ‘Mot s6 quy dinh va han ché cla quéc gia vé sir dung typ huyét thanh khac. Xem cdc yéu cdu hién hanh vé viée chon typ huyét thanh Saimonelia. ) Phat triénld6 duc dug phan loai nhur sau: 0 — khéng phat trirvkhéng duc; 1 — phat trién yéurhoi duc va 2 - phat tridn/cé duc (xem 7.4.2.1, 8.4.1). * — Néu moi trudng dinh duéng dugc sty dung cho hai hoaic ba tng dung khée nhau: it nhat la thy ign phép thir phat trién Salmonella (néu phang thir nghiém thy hign thir vi sinh vat nay). ‘Chon ede chiing theo phuong phap sir dung trong d6 TSA duro ding lam mei trréng ai ching. 76Bang F.1 — Cac vi sinh vat thir nghigm va cac tiéu chi higu nding cia mdi trong nudi cdy thuong ding trong vi sinh vat nude ‘M6l trréng chon ige dé dém vi sinh vit bing cdch so sinh voi mdi trong d6l ching khong chon loc Moitweng? | Tye] Visin vat Tru | chiro a ‘Ching at ching [sé wocw™| Mol [Phuong [Chuan cir] Cac phan ong ate chu’n | nang tradng | phap ‘rung adi kid ching _| ching Collet | t | Evcotwiwnuin | TOWN | Heu | @osaw Ecol coors” | TSA | Binh | F205 | Khudntacmau ving va Coliform ete7-2 | sut | 622°C ooogo wong 6 hugh quang ah voi Jaso 9308- Ecol 2 Kiebsiona ooz0e | TSA | Binh | P2068 | Khudn Igcmau vang ‘pneumoniae tugng khong nhat hon mau cdl miu kien chong 461 voi vikhusn coliform Tink Pseudomonas [0027 no’ | | Bichtinh |Uccrétean] _Khudn ige mau vang ‘chon ee ‘senuginosa® ‘00025; pala @) | nhat hon mau cia miu ibm chang eves [8 Legioneta —[tsori7s1| Higu | 2ngay ains “Legionela oor? | Bcve | Binh | P.205 | Krudntacmau ting. jva11721-2) sult | agayna62)%c | _pneumophita 0180 tugng -xémm-xanh tam-68 ta c& ‘Sngaydén 10 | Legioneta anisa | 00106 iva tron bong dae pty 10822)°C tung Tinh ‘Iroay Enterococcus —_| 00009 hase = | Binh tinh | Ueené toan ~ chong | @6+2)°C faecalis’ ‘0087 phin Pseudomonas | 00028hese| ‘| Binh tinh | Ucchtatn = aeruginosa” 00025, phon ote Ecol” 00012 hose Kimgphin 00013, @ LactoseTIo | S | Ecomiknuin | TovN | Hiu | isan Ecol aoe’ | TSA | Binh | P2058 | Khudnlacmau vang Coliform e197 | sult | casey *c 00012 haga trong mdi xing ob #80 9308- nota mang » Enterococcus | 00175 ‘erogenes Cirobacter reunai | _ 00008 Tinh Enterococeus | o0009heae | ‘| Binh tinh | Uechb etn 7 chon ioe foccats* 00087 phn 0) 80 dee Pseudomonas | oo025nesc| | oinhtinh| ‘| Khun lacmau dé, xenh higu aeruginosa ‘00028 trong mei twdng 8218 NADL ‘102:3 _ Bang F.1 (tiép theo) (Méi trrong* Vi sinh vat Tih Chere u Ching a41 chéng Phuong | Chudn ctr} Cac phan ung age chudn ang phap trong kid _|. chimp mcP \Crostridium perfingans| O8/B/EC | Higu Clostridium ‘Dinh Pa208 | Khudntec mau vang, suét Perfingens lugng dwong tinh voi phép thir phosphatase (2143) (44 1)°C msi ‘teurérng kj Khi Tink Closteium einhtion | —— | Khun taemau xan ‘chon loc bifermentans: Jam, am tinh vai phép: thir phosphatase Bp age Ecol ‘ooo1znosc| | Bintan | echb can = hig 00013, phan (0), Peoudormones Pseudomonas | TOvN | Hisu | (4aanr | Pseudomonas | o0024 | TSA | Binh | P,zos | Khudnigcmaulyclam cN woruginose | 8810] sust | (as+2)"c aeruginosa ‘002s, wong cb huh quang trong 16266) dai UV (360 + 20 nm) coz. Tinh Ecol? ‘cooizhosc| | Binninn | eend cen = chon Ige 00013 hin 0) Fntorocossus | 00008 hase faccalis® 0087 ‘Stanetz va Enterococsi dutng | TCVN Higu (424) hr Enterococcus ‘o000g" TSA Bion | Paz 0,5 | Khudn lacmau dé-nau Bartley ud e102 | suit | (a522)"c recali! ‘c0087 ‘wong sim-néag (80 700 ootre Enterococcus | 00177 facciunt 00178 Tinh E. col” 00012 hose - ‘Binh tinh - - chon loc 00013 Stephyfococcus | 00022 no#e aureus! 0034 8718 NADL ‘9L02:Bang F.1 (tiép theo) Sitsuimt | S | Wikhudniiknikh | TOVN | Higu | casiayny ostium ‘0007 | TsAnose] Sinn | Paz0,5 | Khulnieemaucen ‘Trypose sult ‘sulfil (clostridia) § | 6191-2 | sudt | (a7 £1)"C moi perfingens ‘00080 |mbitrw’ng] twang 5) leso exer, trudng yémm kt tigch mu 2 ge mol ‘wong chan 196 nae ai ve ike iti Tint Eco” [ooot2noge| - [einntnn| — | Khudnige kong od chon toe 00013 mau den, 3c 8. |Costrcnum peringens| 1s0 14188] Higu | @123)h ‘Glosticium 0007 | TsAnaic | inh | Paz05 | Khudnipemau den sudt | (a44-)°C mei | peringens 0080 |miiiyéng} lweng ting kj khi cot7s | tachmau hokeméi trxing chon ioc Jerse i ve} Wisin id i Tinh Bacillus subtilis 00003 - ‘Binh tinh | Uechéteen Pa chon ioc subsp. spizizenii_ phan} M6l trrémg chon Igc dé dim vi sinh vat bing céch 80 s4nh oa mé 4a dure chip nhin truvo a (air dung trong hyp ago bigh, Mottnzong" [Tye] Visinh vat Tidy] chae a ‘Ching ddiching | SéwocM| Mei | Phuong | Chudn co] Cie phan Gng ate ehuin | nang trvdng | phép ‘rung asl | kdb chimg | Colilert t . coliNi Knuan TOWN | Higu (2042)v Ecol ‘001g [ié Collert| Dinh | Pa20,7 [Khudn tac mau vang val ‘color 1a7-2 | suét | (3622)°C ‘00080 auege | heong }<6 hujnh quang 46% v6 laso 9308, inh ola Ecol 2) tnrtic 6 ‘Klebsiella 00208 |Mé Collert| Binh | Faz0,7 | Khudn tac mau ving preumorioe digo | tang {khong nhat hon mau dénh gia cola m&u kiém ching tnrtc a6 i vi vi Kun . . _ color Tinh Pseudomanas | 00027 = [Sinh uns [Veetean| Khudn lac mauvang ‘chon igo] aeruginosa” — | ogc 00125 Frdn() | nhothon may oie my - ching 6z ‘azbe NADL g10z:og Bang F.1 (tip theo} ‘eve! Tegionata[wsott7si] Hieu | 2ngayains Lepioneta DoOF | Meme | Oh ] Paros | Khudn tec mau trang: vatt72i-2] sudt | ngtyrtas:2)°C | aneumaphita aoigo | trwong | wong xémsxanh tama ta ob ‘angeyauita | Loponana ‘on10s | GYPC aa 8 va iron bing dae outst t0 | Lovina (06 core oan anh gi Tink Sngay Enterococcus ‘0009 = | Binhtind | Ge ene - chon ke | @8#2)°C faecalis" | ogc 00087 ‘oan phan @ Pseudomonas 00026 nose] | Binntinn | Uc one = aeruginosa 9025 toan phan Eco 00012 hose otc ting oot phan (0-4) Lactose 776 Ecolmkhén | TCVN | Higu | @1ea)h E col cove | Mémsi | bin | Pe20.7 | Khudnigemauvang Coliform ere | suét | Gei2y%e cootg | trdng | twong trong mei turong cb so 9308. coors | lactose mang 9 Trees Enterococcus | 00175 | quce aerogenes anh gis Citrobacter reunet | _ 0006 Tinh Enterococeus | c0009hoae| —— | Bint tinh | Ure eh = chon bye faecal? ‘00087 toan phan (0). Dodie Pseudomonas [o002shose| | Binhiinh | — Khun ac mau 48, higu jeruginose! Mies xzanh trong moi trzéng mcP \Clostridium perfingens| 9/83EC | Hieu Clostridium: 00007 | Mémdi | Binh | Paz0,7 | Khudn'tac mau vang: sult porfingens cove | tering | tong ‘dung tinh voi phep hosphatase eteqw corza | moP aa ‘thir phosphh (444 1) °C moi enh old Bodie | “MMOH | Crinum | 0007@ | —- | Binhtinn| - | Kudo lacmauxanh hig ‘bitermantans Jam; aim tinh voi phép thie phosphatase Tish E.col# oo01Zhosc] — - | Binhtinn | Ve ché - chon loc 0013, fan phn © $10Z:2Z18 NADLBang F.1 (tiép theo) Pooudomones Pseudomonas | TCVN | Hitu | (aia) | Pseudomonas | 00024 | Momdi | Binh | Paz07 | Khubniacmaulyclam cn soruginasa | 8881 (S0| subt | @e#7°C. ‘oruginesa 000 | tueng | tong 6 hun quang trong 16266) ‘ooss | Pseuso- 10 kun | Kuan tac nhd, phdng suat | (972 1)°C moi iiun?* (eGo tactzin | hoge i, ba mat béng terding hidu Kni | Campytobactercol | goco4. mecca +£.cok | ovo12hose 00013 + Proteus mirabiis | 00023 Tink E.cof? Seoranoge | — | Binhainh] Woche = chon los toan phn i “re (Oytrén Proteus mirabits | 00023 TSA musi! | L | Ecotvixnuin 160 9908-3) Hew aw Ecof 00179 | Chitiét vd phuong phap kid sodt va fou chi chdt trong caliform suit | 4420.8)°C ‘lla mi trvéng MUGIEG neu tong Pry lye E cba 150 9308-31998. mupis® | L | Enterococcinuet | Town | Hu | ~azayny | Enferococcus | 60176 | Chitiétva phurong php kibm soat va iu chi chat wrong ets | suit | aasosy’c Taocalis cia méi trurbng MUDISF néu trong Phu luc E cla 180 enerococcus tres | go177 TOVN 6189-2:2008 (150 7899-2:2000) 7308-1 » Enterococcus | goi78 _ fagciun | Tinh “Aeroceccus virkdans | 00081 hg ge Lactococcustcits | 00016 Staphylococcus | 90192 Z epidermis 8218 NADL S02:Bang F.1 (tiép theo) RVS: L Salmonets TCVN ‘Higu Q4e 3) ‘Saimoneiia - Binh tinh |> 10 khudin] Khudn tec ac trung 977180} subt | 4152 1)°C Enteritidis® ‘90030 lactrén | theo timng mol trréng 19260) Salmonella 00031 XLD hoe | (xem tiéu chudn) ‘Typhimurium* mi tevng ‘Eco | oootz note ee 0013 + Pseudomonas | 00026 nenuginesa Tinh Ecol” 00012 hase - Binh tinh | Ueehétimg - chon lec 00013 phn s 100 Khun tae wen TSA Enterococcus 00009 hodic Dinh tinh |< 10 khuan - faecalis® 00087 fac tron TSA, ‘Mol dng chit ling khBng chen Igo Wai tevdmg? [Tye] Visinh vit Titu | Chie u Ching détchang [Séwocm"] M6 | Phuong [Chuln ec] Gio phan img df chudn nang thing phap frumg Jadicnimg} ibm ching DRCM Lo | Wikehudin kj ehi khir TCVN ‘Hig Clostridium ‘o0007* - Binhtinn | 86 dyc Khuan lac mau den ‘sult (dostrdia) | 6191-1 | sudt otek ae parfingens 00080. 2) 80 8461-9 age | Cad ebeent Eco — [oooiznege[~ — | Biomuinh| 0 aye | Khudn ec khong 6b » higw 00013 (0-4) mau den Nude mudi | ~L | Cécdichiéng pha | TCVN [Dich pha| Eco? | ooo1zncae - Tong 9716180 | leang 0013 Doh pha long a199) Staphylococcus | 00034 fon aureus £30% a ‘thud ung dich mubi 45min abn 1h qa | Dinh | laciTe selena (20 46m 25) °C wang. | (£30 % 26 ‘Dung dich ‘dém ban Ringer (ndng a6 | ‘dauy 114) Dung dich dm phosphet 8218 NADL SL02:8 Bang F.1 (tiép theo) Mi truéng phan lap chon loc Maitraing® [Tye] Visinh vat Tiu | chite u Ching a6i chimg ] 86 wOCM" | _Méi | Phuong | Chudn air] Cio phan dng de chudn ning: trong phap trung 6ichimg} ibm = ching, mCCDA s Campylobacter |1S0 47995] Higu (aaah) ‘Campylobacter 000156 wae) — | Binhitinh| Phat tide | Khudn lac nhd, phdng suat |(41,52 1) °C moi Jejun 00005 tt (2) hoe bi co bé mat bong] |__| tueng hu ht | Campylobacter cot | 0904 _ Tinh +Ecoi | oootainge | - | einntinn | Ueend | hudn gc kntng abe chon lec. 00013 hogic: todn phan trung 00179 hogc. hoge ting 000e0 | phd (0-1) ‘Staphylococcus | 00032 hatic - Binh tinh | Ucehé oan, - _ aurous 09034 pring xD) 3 Salmonella ron | tigu | @aeanr Samonela ‘0021 = [Binh tion | Phat widn | Khuda ob tam mau e7i7(so| sult | 622)"c typhimuctund® binh | den va vidn trong, maw 19250) Seimoneta a teuesng (2 | d6 nnat do sy thay 68: | Enterta”? baa mau ebamol truer Tin Ecol ‘onizneae | — | Dinh tin| Pratuiin | hull tac mau vang chon lee 00013 hose we end ‘tig phn ev) Enterococcus | cooaa hose | — | bin tint | cchd toa z frocaiie! ‘0087 sida ©) ‘Mol wurbng ding cho nhigu mye dich Meitrudng [Typ] Vieinh vat Tit | Chie 0 ‘Ching adi chimg | S8WOCH®| _Mél_| Phuong | chudn ed] Céc phan tng d8¢ chuin | nang trrgmg | phép trang ast | kiém chimg_| chimg Pw 1 | Phaloang db cémut| TCVN | Pha | 45mindén 1h Ecol” opiznoae| TSA | Bin | 230% = cA vi sinh vat 6507 ISO | loang 20°C dén 00013, tong | khsin tacrTo| 6887) 25°C Staphylococcus | 00034 (30% 98 ee eirban Tang sinh sob a8 TeVN Higu (1842) hd Saimonelta 00031 - ‘Binh tinh | 86 dye - phat hign Salmonelia | 9717080] suat | (3642)°C typhismuriam (tay 18250) Selmoneta 00% Entaridi! ee ‘8718 NAD. g107:Bang F.t (két thie) Mat trrong di chimg 61 véi ab dém vi sinh vat ‘Moi truong™ | Typ" ‘Visinh vt Tieu Chere u Chang 46i chimg | S&WOCM®| Méi Phuong | Chudn cir} Cf phan tng dic chuin | ning trying | phép. trung 4éi | kim ching | chimg ecye | S | Bdmkhuiniac [iso1t731| Higu | 2ngdyin “Legionella oore7® | Mémai [ inh | FR20.7 | Krwdn iacmau tring. va ISO sudt ‘Sngay pneumophila buong egng xam-xanh lam-d6 tia 6) ANN734-2 (362.2)°C BCYE da g@ va tron bong dic age ‘ung nh gis sal S| Bémkhudn igo =| Higu | Theo gut cin E ool? coz | Mémti | Binh | Fez0.7 | Khudn lac dé nung sudt_ | trong phurong 0013 | trong | trong theo tig loa php ma TSA cone | TSAas ‘duge ding lam 00179 duge mdi trong 681 | Crasucium 0007 | ean gia chang pertingens Pseudomonas | cova aerugineso Enterococcus | 00087 eacalis “Ten gol by di cia c&c mel turing viet tt néu trong Bang E.2. (Cae ching duge thir aghigm it nhdt ‘Tham khdo bang K8 ching di ching ob 88ntrén http wwe. wee info ve thdng tin 36 lrgng ching cia bo suru tap giéng va cae chi iét en lac; (Ching dug chon tw do, mét trong cdc ching dug sty dung it hat. LL: bi trwbng ling; S: moi rxéng dBc; SS: moi trong ban dB Phat tiBniad dyc duoc phan lagi nhu saur 0 ~khtng phat tién/kong dc; 1 ~phét tién yéulhoi dye va 2— phat tinlod duc (Kem 7.4.2.1, 84.1) Escherichia colf WOCM 00013 duge néu trong tiéu chudin oy thé. Escherichie cof WDCM 00013 sinh f-d-gluconidase manh va WOCM 00202 sinh j-d-gluconidase you Mot 86 quy dinh va han ché cia quéc gia v8 sir dung typ huyét thanh khéc. Kem ede yu chu hign han v8 viée chon typ huyét thanh Salmon. Phat trldndal dye dupe xbp loa l&: O— khng phat tBn/6@ duc; 1 — phat id yéued dye: 2 — phat trib 4699 duc (xem 7.4.1.2: 8.4.1). ‘Nu mdi tnrting BPW durge sir dung cho hai hoe ba ung dung Khe nhau: It nhSt la thye hin phép thi phat tién Salmons {| Chon cdc ching thee phuong phap od sir dung TSA lam mdi trréng dbl ching. L (edu phiong thir nghigm thy ign thi vi sinh vat nay). $8 S10Z:8718 NADLTCVN 8128:2015 Bang F.2 - Cac thuat ngir viét tat vé méi trrdng dug sty dyng trong Bang F.1 glycin, vancomycin, polymyxin B, eycioheximid Thugt ngte moi trong Ten day di cia méi trong Tieu chudn jet tat BcYE Moi truéng chat chiét ndm men 6ém than ISO 11731 va ISO 11731-2 Botton Canh thang Bolton 180 17985 BPW Nude dem pepton TCVN 6507 (ISO 6887), TCVN 9717 (IS 49250) DROM ‘Moi trudng phan bigt b8 sung clostridium TCVN 6191-1 (ISO 6461-1) Gvec ‘Thach Gem than chat chiét ndm men bd sung 180 11731 va ISO 11731-2 ‘Thach clorua triphenyiltetrazolium lactose: bb sung nati heptadecytsulat TCVN 6187-1 (ISO 9308-1) ‘Thach deoxycholat cefoperazon than cai bién ISO 17995 ‘Theich perfringen Clostridium ding cho phurong phap loc 98/83/EC TOVN 6189-1 (ISO 7889-1) MUDISF ‘Moi trudng 4-metylumbellferyt-a-D glucoside isF MUGIEC Moi truéng 4-metylumbellfery+3-D glucuronide 180 9308-3 rec Preston anh thang Preston 180 17995 Pseudomonas CN ‘Thach axit nalidixic xetrimid Pseudomonas TCVN 8881 (ISO 16265) Rvs Canh thang Rappaport-Vassiliadis pepton cau tuong TCVN 9717 (ISO 19260) Stanetz va Bartley Mol truéng Slanetz va Bartley ‘TCVN 6189-2 (ISO 7899-2) ‘Sat sulfit Thach sét sutfit TCVN 6191-2 (ISO 6461-2) Tryplose sulft (TS) ‘Thach tryptose sulft TCVN 6181-2 (ISO 6461-2) TSA ‘Thach trypton dau twong - Tsc “Thach sulfit xycloserin tryptose (khong chira Iso 14188 long 08 irteng) xto ‘Thach deoxycholat lysin xylose TOWN 9717 (ISO 19250) YEA Chat chidt nim men 180 6222TCVN 8128:2015 Phy luc G (Quy dinh) ‘St dung céc bidu 43 kiém chteng dé kiém soat phép thir dinh lrgng méi trvéng nuéi cdy dac G1 Yéu cau chung Phy luc nay m6 ta vibe str dung bidu ad kiém ching dé theo dai cdc két qua, dc biét fa két qua biéu thi ty I@ higu suat, Pp, nhur chi tiét trong 7.2, khi thir nghiém thach chon lec hodc khéng chon Igc dya vao thach déi chtrng khéng chon gc hoc cdc mé thach chon I9c tuong ty da duge ch4p nhan. Can cha y khi siv dung bigu d& kiém ching a8 thir nghiém gitra céc mé cilia cing loai thach, nhu bat ky ‘sy suy giém chat lung nao olla cae ma lin tiép 66 th8 khéng duge 16 rang, triv khi ¢é¢ dung dich huyén phil thy nghiém urge kiém chteng cn than 48 cho so iwgng sinh vat thich hyp, hodc stk dung RM (xem 3.4.6). Bat ky hé théng kiém soat chat lugng mdi trvéng nao dia vao viée kiém tre gifra cac mé phai duge xéc nhfn la phi hop v6i myc dich truréc khi dura vao sir dung. M&i ngudn gc va mé thach thi nghigm d&u cho thay mirc 49 khdc nhau vé higu sudt va céc phong thd nghiém riéng ré c6 thé su dung cac thach &4i chieng khac nhau dé so sénh vi thach thir nghiém. Do 6, cdc phang thir nghiém ring ré phai thiét lap va ching minh c&e giéi han clia minh vafhoge Kho&ng hé sé higu suét od thé duoc chp nhén déi véi mai loai thach thir nghigm si» dung thudng xuyén trong khoang gid tri hé s6 higu su&t quy dinh trong 7.2.2.1.2. Cac biéu dd kiém ching duge chun bi tir cae d@ ligu danh gia x4c nhén ban dau, thiét lap cdc giéi han chdp nhan sir dung phan tich théng ké va sau 6 cc bidu dd nay duge sir dung 48 theo d6i cde mé thach tiép theo, Thuong xuyén danh gid cdc gidi han (vi dy ct? sau méi 30 phép thit) va didu chinh khi can (théng tin bb sung duge diva ra trong G.2.6). Cc quy trinh yu cu sir dung dung dich huyn phi: vissinh -vat:cé. néng:46.48 biét cla ching dich quy inh cho méi trong thir nghiém néu trong Phy luc E hoc Phu luc F; day 1a gid tri dich d6i voi phép thir, Huy&n pha thir nghiém la RM thuong mai (xem 3.4.6) ho&c duge phéng thir nghiém chun bi tir cc giéng lam viée 4 chudn héa ky cla cdc chting déi ching. Nang 46 vi sinh vat trong huyén phis phdng thir nghiém (gid tri dich) phai cho thy 8n dinh va ddng nhét trong sudt théi gian sir dung. Bé thi wu hoa viéc kiem soat, cdc huyén phi phai chira xp xi 100 cfu (khoang 80 cfu t6i 120 cfu) trong mot thé tich dich cy dure Gira lén dia thach va thyéng thyc hién céc phép ther it nhdt hal lan Kap lai, Bang 1 (xem §.4.2.5.1) cung cép cae gid tri v8 d6 chum & cdc ndng a6 dich cdy khdc nhau cho thdy ‘t&m quan trong ctia viée duy tri dich cdy téi wu nay. a7TCVN 8128:2015 Cae dia thach phai durge cy bang cdc kj thuat dan bé mat, dé dia hoae lec mang, thich hop voi phuong phép chudn ai chiing ma thach duge sir dyng va duge Ui trong cac diu kién quy dinh tai ce tiéu chudn oy thé 46. Cac khudn ige ¢6 mat trén hod trong mdi éta phai dee dém phii hop voi cdc phuong phap chun a6i chting va céc hé sé hiéu suat tinh duge trong 7.2.1.1. G.2 Sir dung biéu dé kiém chieng G.2.4 Yéucdu chung ‘Dé thiét lp mét bidu 48 kiém cht¢ng méi, cn thyc hign it nhat 10 phép thir trén cdc mé khac nhau cla cling loai thach thir nghiém, tét nhdt la thyc hién fp lai hai ln va vao nhiing ngay khac nhau do cdc ngudi khac nhau thyc hién (cdc didu kign t4i lap néi phdng thir nghigm). Sir dung 20 phép thir (nh trong G.2.2) $8 cho gidi han dang tin cay hon so véi biu 43 dug chuan bj tir 10 phép thi ban dau (s6 long téi thidu) phai duge tinh lai Khi 6 s&n 20 k6t qua. Phai duy tri cdc bidu 66 riéng ré theo mét day it nhdt 30 phép thir dugc gitr lai cho méi dénh gid chét lvgng méi trvdng (xem G.2.6). G.2.2. Dyng bidu 43 kidm chéeng Quy tinh nay da trén dich dy chudn c6 chiva (100 + 20) cfu trong 0,1 mi dich cdy duge st dung a6i V6i phép thir sir dung ky thuat dan dla quy trinh nay. CCing thich hop khi sir dung cdc thé tich khéc cla dich cy d6i v6i cdc kj thual khdc nhu &é dia hoae loc mang, voi didu kién la dich cdy cé chia s6 Iwgng khudn lac nam trong dai ch4p nh€n (100 + 20) cfu. ‘Trung binh cilia hai day sé am lap lai trén phép thir thie /a x cfu/0,1 ml ai voi moi truong thir nghiém va y, cfu/0,1 mi déi vei méi trxéng abi chimg. Khi 46, hé s6 higu sudt ca phép ther thir fla xi/ yi = 6. Déi voi cdc day phép thir tir i= 1, 2, ...0, trong dé 7 1a sé t6i thidu 10 va t6t nhat la 20, sir dyng Cong ‘thie (G.1) 48 tinh gia tri trung binh, 7 cia Pr: (G.1) ‘St dung Céng thire (G.2) 48 xéc dinh céc gid tri trung binh, R olla Pr: (G2) “@- adh ~ Trong a6: i 86 phép thir; 1 la tbng 36 cde phép thir, 1 la gid tr Pp thir i, 88TCVN 8128:2015 X€c dinh cdc 46 I8ch chudn (s) nh sau: R =09065xR= Boos 1128 (G.3) CHU THICH: Hing 66 0,8865 (node nghich do cia 1,128) lb chun dug ASTM khuyén cBo a8 xae Ginn a9 Igch chun tir dai trung binh cc gié tr thir nghiém lap tai hal lan. Tinh cdc-gid tri gidi han 95 % (+ 2s) va 99% (43s) cila sy phan bé cdc két qua. Trén truc ¥ ola biéu 4d kiém ching, danh dau cac vj tri cia gid tr] trung binh téng thd Pp, 7, tng tri cla hai gid tri trén (+2s va +3) va hai gia tri durdi (2s va -3s), sau da vé cdc duéng song song voi frye X. Dung cae gid tri Pa thir 1, 2, ... ntrén truc X (xem vi dy). MGi khi chun bj mé méi turéng méi, cn thiy nghiém st? dyng huy&n phi dich cdy chudn va Pr tinh dugc tir ty $6 cita sé cfu dém dugc sau khi ti Am trén méi irréng thir nghiém va mdi trudng a4i ching nu én, Glé-tt nay due vé trén biéu a3 kiém ching va duge kiém tra lai dyra vao cdc giéi han thu duge. G.2.3 Vidy vd dyng bidu ad kiém chieng sir dyng 20 két qua Bang G.1 cho cac két qua tir 20 [An kiém tra hé sé higu sudt lién tiép trén cdc mé cla cuang loai thach thir nghiém khéng chon loc si* dung dich cdy chu&n 110 cfw0,1 mi d&m duge trén méi trrong déi chirng khéng chon loc (s6 dém chinh xéc trén méi trudng di ching $6 khéc so voi sé dém duge trong thye t8, nhung s6 ligu duge ding dé dua ra nguyén tic cla biéu dd kiém ching 4 chi tir cdc hé 56 bigu suat tinh due). Bang G.1 ~ Bao cao kiém tra hé sé higu sudt cia 20 ln lién tidp trén thach khdng chon loc ding 48 xay dung bidu a8 kiém ching (cho cdc s6 dém thye chi trén mél trong thach thir nghigm, cdc sé dém trén méi trudng 4éi ching -y; 1a 110 cfu) Kt qua s6 phép thir nghigm () fs a} 2;sf4{]s]6f]7] 8] s | Shu (thach thirnghigm-x) | 95 | 102 | 94 | o7 | 105 | 62 | 98 | 105 | 103 | 116 h _ o.26 | 0.93 | 0.85 | 088 | 0,95 | 062 | 0,89 | 0.95 | 095 | 4,05 - | 097 | 0,08 | 003] oo7 | - | 0,06 | 0,08 | o01 | 011 is a [12 | 13 | 44 | 1s | te | 17 | 18 | 19 | 20 Cfu(thach thirnghigm-x) | 95 [| 90 | 89 | 116 | 114 | 110 | 114 | 98 | 88 | 102 4 086 | 082 | 081 | 1,05 | 1,04 | 1,00 | 1,04 | 089 | 080 | o93 | 0,19 | 004 | 0.01 | 024 | 9.01 | 0,04 | 0,04 | 0,15 | 0.09 | 0,13 89TCVN 8128:2015 Can twu ¥ rang két qua duéi cla phép thir 6 6 trong vi du nay cla thach khéng chon Igc thép hon khodng cho phép di véi gia tri ch4p nhan Pp tir 0.70 dén 1,40 clla thach khéng chon loc (xem 7.2.2,1,2) va tat ca cdc két qua nhu thé dug coi [a ngoai Ié trong vie thu ldy 46 Iéch chuan va gid tri trung binh d6i véi biéu dd kiém ching. Cae ly do két qua thiy nghigm PR nam ngoai dai cho phép cn duge nghién ctu vi ching thung lién quan dén vige thye hign khong t6t quy trinh thiv nghiém chir khéng phai do chét lugng méi trvdng kém va thuréng tim durge khi dich cdy duge str dung nm ngoai khodng 6 chym quy dinh tiy 80 ofu abn 120 ofu. Gia tri Pa trung binh bang voi 1,110 = (0,86 +0,93 + 085 +... 40.80 + 0.93)/19 = 175/19 = 0.92 (G4) (thé hign bang duéng ké dam trén Hinh G.1) Khoang céc gia tri (R = |r, -1.,|) duroe xde dinh I& chénh lech tuyét 461 cla cdc gia tri lién tiép, ngoai tri? gid ti bj loai (vi dy: phép thir sé 6 trong Bang G.1 & trén), nghia la: 0,93 — 0.86 = 0,07; 0,93 - 0,85 = 0,08; 0,88 — 0,85 = 0,03 wv... ‘Gid tr trung binh bang: = DR Mn-1)= (0.07 +008 +003 0,09 +0,13)/18 = 1.43/18 = 0,08 (G5) Do 68, 49 Iéch chudn, s = 0,8865 x 0,08 = 0.071 Cac gidi han tin cay 95 % (thé hign bang dung ké chdm nhat trong Hinh G.1) Ia: 0,92 £ 2x 0,071 = 0,92 + 0,14 = 0,78 dén 1,06 (G6) Cac gidi han tin cay 99 % (thé hién bang dudng ké chdm dam trong Hinh G.1) 1a: 0,92 + 3x 0,071 = 0,92 + 0,21 = 0,78 dén 1,13 (G7)TCVN 8128:2015 ‘CHU DAN: Y hi s6 higu sub, Pe X 86 phép thir Hinh G.1 — Bidu 48 kiém chieng duge xay dyng tir cdc hé sé higu sudt thu duge ti 20 ln kiém tra dau tién néu trong Bang G.1 (44 loai triv két qua ngoal Ig sé 6) G.2.4 Banh gid higu nang va gidi thich két qua Mé méi truéng nudi cdy sé diye chap nhan néu cae tiéu chi chat lugng vi sinh In tiéu chi chung (xem 6.2) dau dap tng. Mé mdi trong mdi bi loai bé néu cac két qua sau day xudt phat tir cdc phép thir dinh Igng néi trén khéng kiém soat duge nhu sau: — vipham gidi han + 35; ~ hai trong ba quan sét trong hang ngang vugt qua giéi han cnh bdo + 2s; = sAu quan sat trong hang ngang la tang déu hoae giam déu; — chin quan sat trong hang ngang trén cing mét bén clia gia tri trung binh. CHU THICH: Ti chi eda bén quan sat trong hing ngang vet qué mic # 1s cing cho thy ob vin ab. 1TCVN 8128:2015 G.2.5 Cac cach tiép can khac 48 ther higu nang cla méi trong Quy trinh si dung bigu 48 kiém ching 48 ther higu nding otia méi trong néu trén thu duc 48 dung 4 thi cac gia tri Pa ma khéng c&n chuyén logis cla s6 dém khuan lac. Nhing phuong phdp thay duoc ch4p nhn dura trén dé thi tre tiép cdc s6 dém khudn lac hoac s6 dém Khudn lac chuyén vé logio, néu cho thay thich hyp. Céc phuong phép kiém tra xem sy phan bd dir figu 6 dém khudn lac ¢6 phi hgp voi phan b6 chudn hay khdng khi khong chuyén vé logyo Si’ dung phép thiv Kolmogorov-Smimov ho&c phép thir khi-binh phuong, ede vi dy dirge dura ra trong NEN 6603. [39] Tuy nhién, can wu y rang néu cdc s6 dém khéng bao gdm hé sé pha long, khi 66 khéng chac bién 481 vé logie sé 1a thich hop. Viée chuy&n déi tru tiép s6 d&m (vi du 100 cfu/dia) cdn gid dinh phan bd Poisson 8 chuyén adi sé dém x la Vx . Ngoai ra, trong tinh huéng bat ky khi s6 éém khuan lac dugc dung dd thi tryc tiép thi cn dam bdo rang mire dich cy thir nghiém la khong déi gifra cdc phép thts, neu. khong kétqud thu duge sé bi sai G.26 Xem xét dinh ky cdc biéu dé kiém ching Cac bidu 48 kiém ching dau tién va tiép theo déi voi mét méi trweng thir nghiém phai duge xem xét inh ky 48 dam bao rang cAc gidi han thiét Iép van con chdp nhan duoc. Biéu 4d dau tién chira tdi thigu la 20 diém do liéu duoc xem xét dé thidt lap gidi han ban dau va sau 46 cac biéu dd tiép theo c6 thé duoc xem xét tai cdc tn suat du kién cho méi 30 aiém dor ligu bang cac quy trinh dudi day. Mot khi biu a8 kim ching durge hoan tat, tinh lai gia tri trung binh va dé léch chudn s, bd qua moi két qua nim ngoai dai ch4p nhén hodc gidi han thiét lap. Trudng hyp si dung viée chuyén adi vé logso, thi thyc hién tat cd cde phép tinh si dung két qua chuyén ai logo. So snh 66 léch chuan cia bidu dd hign hanh voi 66 Kéch chudn cila t&t c cdc két qua trude 46, Sis, va kim tra bign thign st dung cdc tiéu chi sau day: re Se

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