THC HNH VI SINH I CNG

BO CO THC HNH MT S PHN NG SINH HA

1. KH NNG CHU MN Nhm xc nh kh nng chu mn ca vi sinh vt. Mi trng: NA: Nutrient + Agar + Nacl 8.5% Kt qu: nhng vi sinh vt c kh nng chu mn s tn ti trong mi trng, ngc li s cht. 2. TH NGHIM BNG UREASE NGUYN TC Xc nh kh nng ca vi sinh vt tng hp enzyme urease xc tc s thy phn urea to ra 2 nguyn t NH3 v CO2 lm tng pH ca mi trng v c th c theo di qua s thay i mu ca cht ch th pH. Th nghim urease l c trng cho cc loi Proteus spp. V thng c dng phn bit cc dng Proteus vi cc thnh vin khc ca Enterobacteriaceae. C s sinh ha: (NH2)2CO + H2O 2NH3 + CO2 tng pH mi trng phenol. MI TRNG Th nghim Urease c thc hin trn 2 mi trng: Mi trng urea lng Rustigian-Stuarts Urease Broth Mi trng Christensen Urea (mi trng thch nghing) KT QU: Mi trng urea lng Rustigian-Stuarts Urease Broth: Dng tnh (+): Mu tm trong khp mi trng m tnh (-) : Mi trng khng c s i mu (mu vng cam) Mi trng Christensen Urea: Dng tnh (+): Mu tm trn b mt mi trng thch nghing + + + + : ton b thch i mu + + + : ch mt thch i mu
1 TRN THANH TNG MSSV: 10243221| Thy: TRNG V ANH DNG

THC HNH VI SINH I CNG

+ : mt thch nh i mu, phn mi trng cn li khng i mu

(+) nhanh: mu i trong vng 1-6 gi (+) chm : mu i trong vng 24 gi n 6 ngy hoc lu hn. 3. TH NGHIM GELATINASE MC CH Th nghim kh nng phn gii gelatin bi gelatinase C S SINH HA
gelatinase

Gelatine

polypeptide + acid amin

Gelatine trong mi trng dinh dng mi trng ng c VSV phn hy gelatine mi trng lng. i chng dng: Aeromonas hydrophila i chng m: E.Coli Mi trng s dng: Nutrient Gelatine Dng ng nghim thch su Cy vi sinh vt v nhit phng. KT QU: c kt qu (+): mi trng tan chy (-) : mi trng khng tan chy

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

THC HNH VI SINH I CNG

4. TH NGHIM AMYLASE MC CH Pht hin cc vi sinh vt c h enzyme amylase Dng thuc th I2 nhn bit. Nu trong mi trng th nghim, vi sinh vt c h enzyme amylase, amylase s thy phn tinh bt thnh nhng cu t nh hn, khi cho I2 vo, dung dch c mu tm xanh (+). 5. TH NGHIM KIA/TSI MC CH Pht hin kh nng: s dng cc ngun cacbonhydrate sinh H2S to hi (gas) MI TRNG: KIA: Kligler iron agar Pepton Lactose Glucose NaCl Feric ammonium citrate Sodium thiosulphate Agar Phenol red Nc ct pH 7,40,2 Mi trng KIA ch c hai loi ng 0.1% glucose, 1% lactose. Mi trng TSI c thnh phn ging KIA, c b sung thm 1% sucrose. 20g 20g 1g 5g 0,5g 0,5g 15g 0,025g 1 lt

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

THC HNH VI SINH I CNG

KT QU:

370C/24h

Quan st: Phn nghing / phn su / hi / H2S

C ch: C ba trng hp xy ra: Ch s dng glucose, sau 18-24h nui cy phn nghing (b mt) tr nn c pH kim v phn ng (phn su trong ng nghim) c pH acid. Do Glucose trn b mt ca mi trng c vi sinh vt oxy ha hon ton thnh CO2 v nc thu ly nng lng, p ng nhu cu nng lng trong tng trng. Vi sinh vt tip tc d ha peptone qua gii phng NH3 lm phn b mt ca mi trng c pH kim. Trong khi , phn sau trong mi trng c iu kin oxy khng y , glucose c ln men k kh sinh cc acid hu c lm pH mi trng gim. S dng c glucose v Lactose, sau 18-24h, ton b mi trng u tr nn c PH acid v s bin dng ng thi c hai loi ng gip vi sinh vt nng lng tng trng m cha cn s dng n peptone. Nu ko di thi gian nui cy qu 24h b mt pH s tr nn kim do ht ngun cacbon v vi sinh vt pha s dng n peptone. Khng s dng glucose, lactose: vi sinh vt s bin dng peptone thu ly nng lng v vt cht cho s tng trng. tuy nhin do peptone ch c bin dng trong iu kin hiu kh nn hin tng kim ha ca mi trng ch din ra trn b mt mi trng. Kh nng sinh H2S do trong mi trng sodium thiosunfat, vi sinh vt kh sunface c th kh cht ny, nh c enzyme thiosunfat reductase gii phng H2S, H2S s phn ng vi Ion Fe2+ ca ch th amonium citrate to kt ta mu en F2S.

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

THC HNH VI SINH I CNG

Mt s trng hp c th xy ra khi dng mi trng KIA/TSI

10

11

ngha: nh danh cc loi vi khun c kh nng: s dng cc ngun cacbon khc nhau v c kh nng sinh H2S, kh nng sinh hi. Thng c dng nh danh cc vi khun gram (-), vi sinh ng rut. 6. TH NGHIM KH NNG SINH H2S MC CH: Pht hin kh nng sinh H2S ca vi sinh vt. C S SINH HA:
desulfohydrase

Acid amin cha S

Thiosulfate reductase

H2 S

Thiosulfate

H2 S

H2S sinh ra c nhn bit bi ion st, ch to kt ta mu en (FeS, PbS). MI TRNG: KIA, TSA (thch nghing) SIM, PIA (thch su) BSA (thch a) Cy vi sinh vt ln mi trng, 37oC, 24 - 48h
TRN THANH TNG MSSV: 10243221| Thy: TRNG V ANH DNG

THC HNH VI SINH I CNG

KT QU: Xut hin mu en trong mi trng (+) Khng xut hin mu en trong mi trng (-)

7. TH NGHIM CATALASE MC CH: Pht hin vi sinh vt c h enzyme catalase C S SINH HA: Catalase hin din cc VSV hiu kh v k kh ty
Catalase

H2 O2 (hydrogen peroxide) THC HIN

H2O + O2 (bt kh)

VSV ly t mi trng nui cy (lng, rn) t VSV ln lam knh sch Nh H2O2 30% Quan st sau 1-2 giy Phn ng (+): c bt kh xut hin

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

THC HNH VI SINH I CNG

Phn ng (-): khng c bt kh xut hin

Th nghim catalase trn a petri: s dng H2O2 30% 8. KH NNG LN MEN CC LOI NG
7 TRN THANH TNG MSSV: 10243221| Thy: TRNG V ANH DNG

THC HNH VI SINH I CNG

NGUYN TC: Xc nh kh nng ca mt s vi sinh vt ln men (phn gii) mt carbohydrate c hiu c trong mi trng c bn tng trng. Khi s dng cc ngun carbohydrate ln men, ty theo phng thc ln men cc sn phm c to ra s khc nhau: ru, cc acid hu c, CO2 Trong cc trng hp, cc sn phm to thnh u lm gim pH ca mi trng. CO2 c to thnh s c by li to thnh bt kh trong ng chung Durham lm ni ng chung ny (trng hp s dng mi trng lng) hoc lm v thch khi cy trong mi trng thch su (trng hp s dng mi trng rn). Vic gim pH th hin qua s thay i mu ca cht ch th pH c trong mi trng. Th nghim kh nng ln men thng c dng nh danh cc vi sinh vt ng rut (Enterobacteriaceae c kh nng ln men glucose, E.coli, Klebsiella, Enterobacter ln men c gluocose v lactose) v phn bit mt s ging vi sinh vt.

MI TRNG: Phenol Red Broth Base THC HIN: Cy vi khun mi hot ho vo cc ng nghim, t 36 0C, theo di hin tng sinh acid sau 1-3 ngy. - Lc v trng mi trng qua mng lc c kch thc l 0,45m hoc 0,2m. - Phn phi vo cc ng nghim . Hp 1210C trong 15 pht - Tin hnh cy chng vo cc ng mi trng bng que cy vng. - cc ng mi trng 370C trong 18 20 gi. Trong trng hp khng nh kt qu m tnh, cn dng paraffin bt kn nt bng v theo di thi gian c th ko di n trong 14-30 ngy.

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

THC HNH VI SINH I CNG

KT QU:

Phn ng (+): mi trng chuyn mu vng Phn ng (-): mi trng c mu

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

THC HNH VI SINH I CNG

10

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