Bi tiu lun

TRNG I HC DUY TNKHOA DC

BO CO THC HNHMN : VI SINH CN BNM MN HC : MIB 251

Gio vin hng dn :L Th MaiLp mn hc:MIB251F2Nhm:3Nhm trng : - Phan Vn SThnh vin : - Vn B Bnh Nguyn Nguyn Th Thng Thng Nguyn Th Tnh Nguyn Th Ngc

Nng,thng 5, nm 2015.Bi 1. CC LOI HNH TH CA VI KHUN.Theo hnh thi b ngoi,vi khun thng c chia thnh 3 loi hnh th chnh: cu khun,trc khun v xon khun.1. Cu khun :(Coccus - t ting Hy Lp Kokkos - ht qu): l loi vi khun c hnh cu. Nhng c nhiu loi khng hn hnh cu th d nh hnh ngn nn nh ph cu khun - Diplococcus pneumoniae hoc ht c ph (lu cu khun Neisseria gonorrhoeae).Kch thc ca vi khun thng thay i trong khong 0,5x1,2m (1 = 10-3 mm). Tu theo tng loi m chng c nhng dng khc nhau.c tnh chung ca cu khun:- T bo hnh cu c th ng ring r hay lin kt vi nhau.- C nhiu loi c kh nng gy bnh cho ngi v gia sc- Khng c c quan di ng.- Khng to thnh bo t.

a. Ging Monococcus (n cu)Thng ng ring l tng t bo mt, a s chng thuc loi hoi sinh. Thng thy chng sng trong t, nc v trong khng kh (Th d nh Micrococcus agilis, M. roseus, M. luteus)

b. Ging Diplococcus (song cu) :(T ting Hy Lp - Diplos - thnh i) phn cch theo mt mt phng xc nh v dnh nhau thnh tng i mt. Trong s ny c mt s c kh nng gy bnh nh ging Neisseria - Lu cu khun N meningitidis gonorrhoeae - No m cu khun).

c. Ging Tetracoccus (T cu):Thng lin kt vi nhau thnh tng nhm 4 t bo mt. Chng thng gy bnh cho ngi v mt s c th gy bnh cho ng vt.

d. Ging Streptococcus(Lin cu)T ting Hy Lp (Streptos - chui) chng phn cch theo mt mt phng xc nh v dnh vi nhau thnh tng chui mt di. Th d nh Streptococcus lactic. Strep-pyogenes.

e. Ging SarcinaT ting Hy Lp Saricio - gi hng. Phn cch theo 3 mt phng trc giao vi nhau, to thnh nhng khi t 8 - 16 t bo (hoc nhiu hn na). Trong khng kh chng ta thng gp mt s loi nh Sarcinalutea, Sarcina auratiaca. Chng thng nhim vo cc mi trng trong phng th nghim v to thnh mu vng.

f. Ging Staphilococcus (T cu)

T ting Hy Lp Staphile - chm nho. Thng chng lin kt vi nhau thnh nhng m trng nh chm nho. Chng phn cch theo mt mt phng bt k v sau dnh li vi nhau thnh tng m nh hnh chm nho. Bn cnh cc loi hoi sinh cn c mt s loi gy bnh ngi v ng vt (Staph. Curcreus, Staph. Emidermidis ...)2. Trc khun : L tn chung ch tt c cc vi khun c hnh que. Kch thc ca chng thng t 0,5 - 1,0 x 1 - 4 m.Thng gp cc loi trc khun sau y:a. Bacillus (Vit tt l Bac)

trc khun gram dng, sinh bo t. Chiu ngang ca bo t khng vt qu chiu ngang ca t bo. V th khi to thnh bo t t bo khng thay i hnh dng chng thng thuc loi hiu kh hoc k kh khng bt buc.

b. Bacterium (vit tt l Bact)Trc khun gram m khng sinh bo t. Thng c tin mao mc xung quanh t bo ngi ta gi l chu mao. Cc ging Salmonella, Shigella, Erwina, Serratia u c hnh thi ging Bacterium.

c. Pseudomonas (vit tt l Ps)Trc khun gram m, khng sinh bo t, c mt tin mao (hoc mt chm tin mao) mt u. Chng thng sinh ra sc t. Cc ging Xanthomonas. Photobacterium, Azotomonas, Aeromonas, Zymononas, Protaminobacter, Alginomonas, Mycoplazma, Halobacterium, Methanomonas, Hydroginomonas, Carloxydomonas, Acetobater, Nitrosomonas, Nitrobacter u c hnh thi ging Pseudomonas.d. CorynebacteriumKhng sinh bo t, hnh dng v kch thc thay i kh nhiu. Khi nhum mu t bo thng to thnh cc on nh bt mu khc nhau. Trc khun bch cu (Corynebacterium diphtheriae) c bt mu hai u lm t bo c hnh dng ging qu t. Mt s khc c hnh thi ging Corynebacterium gm c Listeria, Erysipelothric, Microbacterium, Cellulomonas, Arthrobacter.

e. Clostridium (Vit tt l Cl, ting Hy Lp Kloster - con thoi)Thng l trc khun gram dng. Kch thc thng vo khong 0,4 - 1 x3- 8Sinh bo t, chiu ngang ca bo t thng ln hn chiu ngang ca t bo, do lm t bo c hnh thoi hay hnh di trng.Chng thng thuc loi k kh bt buc, c nhiu loi c ch. Th d nh cc loi c nh nit. Mt s loi khc gy bnh. Th d vi khun un vn ... Cl. Botulium3. Vi khun hnh xon: gm 3 dng sau :a. Phy khun: L tn chung ch cc vi khun hnh que un cong ging nh du phy. Ging in hnh l ging Vibro (T ch La tinh Vibrare - dao ng nhanh). Mt s ging phy khun c kh nng phn gii xenluloza (Cellvibrio, Cellfalcicula) hoc c kh nng kh sunfat (Desulfovibrio).b. Xon khun:Gm cc vi khun di,mnh,c nhiu vng xon.Mt s c kh nng gy bnh cho ngi nh xon khun giang mai(Treponema pallidum),xon khun Leptospira,xon khun hi quy(Borrelia).

c. Spirillum - T ch Spira - Hnh cong, xon gm tt c cc vi khun c hai vng xon tr ln. L loi gram dng, di ng c nh c mt hay nhiu tin mao mc nh.a s chng thuc loi hoi sinh, mt s rt t c kh nng gy bnh (SP. Minus) c kch thc thay i 0,5 - 3,0 - 5 - 40.Hnh nh quan st thc nghim :Mu ly t cc mng bm trn rng.

Bi 2 : PHNG PHP NHUM VI SINH VT*NGUYN TC:Nhum mu l qu trnh lm cho t bo hoc cc thnh phn ca t bo vi sinh vt c mu di tc dng ca cc loi thuc nhum.*MC CH Gip vic quan st hnh dng t bo, cc thnh phn cu trc t bo ca vi sinh vt c d dng. Gip phn bit cc chng vi sinh vt vi nhau do vic bt mu khc nhau i vi cc loi thuc nhum to iu kin cho vic phn loi, nh dng vi sinh vt .*PHN LOINhum mu vi sinh vt

Nhum khng c nhNhum c nh

Nhum nNhum kpNhum tin maoNhum bo tNhum Ziehl - NeelsenNhum Gram

bi ny s tm hiu phng php nhum n v nhum Gram thuc phng php nhum c nh.A. DNG C, MI TRNG V HA CHTCc dung dch nhum gm: Dung dch xanh methylene. Dung dch tm kt tinh ( Crystal violet ). Dung dch lugol. Dung dch ty mu. Dung dch nhum b sung ( Safranin O hoc Fuchsin ).B. CCH LM TIU BN 1. Cch ly vi sinh vt lm tiu bn:Mun lm bt k mt loi tiu bn no v vi sinh vt cng phi thc hin ccthao tc ly mu vi sinh vt lm vt bi trn tiu bn. Cc thao tc nh sau:- t n cn.- t ng nghim mu hay a Petri c vi sinh vt vo gia ngn ci v ngn tr ca bn tay tri,lng bn tay nga ra. ng nghim hi nghing nhng khng c canhtrng vi sinh vt chm vo nt bng i vi ng nghim.- Tay phi dng que cy v v trng bng cch h trn ngn la n cn. que cy thng ng trn ngn la n cn cho n khi u que cy nng ri t tt v di chuyn que cy theo chiu nm ngang trn ngn la.- Kp nt bng vo gia ngn t v lng bn tay phi, xoay nh nt bng 1vng v ko nt ra. Gi nguyn nt bng nh vy cho n khi y nt vo.- t ming ng nghim trn ngn n cn.- Kho lo a que cy ( ngui) vo ng ging ly mu v nh nhnga que cy ra.+ Nu ng mu l mi trng lng ch cn nhng u que cy vocanh trng ri rt ra.+ Nu ng ging l mi trng c th dng que cy ly mt cht sinhkhi vi sinh vt trn mt thch(nu quan st mt loi vi khun th nn ly trong mt khun lc) v ho u vo git nc ct v trng trn phinknh. Ch thao tc ht sc nh nhng ly mu m khng cy mt thch ln.- Rt que cy ra, t ming ng nghim, y ng nghim nghim li v ng vo gi.- a git canh trng (hoc sinh khi) vi sinh vt u que cy t vogia phin knh lm vt bi.- Kh trng li que cy trn ngn n cn ri ct vo gi.2. Lm vt bi:t mt git nc sch vo gia phin knh, sau dng que cy khtrng ly mt vng que cy vi sinh vt cho vo git nc v dn u ra. Trng hpnui cy trn mi trng lng th dng pipet ly dung dch nghin cu. H khphin knh c vi sinh vt trn ngn la n cn hoc kh t nhin ta c mtvt m trn phin knh gi l vt bi.Ch :- Lng vi sinh vt ly va phi.- Vt bi trn, gn, tht mng.- Cc t bo vi sinh vt c dn u, d quan st.C nh vt bi:- Vic c nh vt bi nhm cc mc ch sau:+ Git cht vi sinh vt chng d bt mu v an ton khi tip xc (nu l visinh vt gy bnh).+ Gn cht vi sinh vo phin knh lc nhum khng b ra tri.- Cc cch c nh:+ C nh bng nhit: dng kp g kp phin knh, h mt di caphin knh qua li trn ngn la n cn 2 3 ln (chng vi ba giy), trnh khng tiu bn qu nng.+ C nh bng ho cht:* Nhng vt bi vo cn 95o khong 10 15 pht ri sy kh.* Ngm trong axeton 5 pht.* Sy bng hi formol 10 15 giy.C. PHNG PHP NHUM:1. Phng php nhum n:Nhum n l dng 1 loi thuc nhum nhum. Cc loi thuc nhumthng dng nhum n l:- Fuchsin kim.- Xanh metylen.- Tm gentian.- trung tnh.Phng php nhum: t tiu bn c nh ln gi ri nh vi git thucnhum ph kn vt bi. Sau 1 2 pht, thuc nhum i v ra nh bng ncsch ti khi no thy nc ra chy qua vt bi khng cn mu na l c. tiubn kh dn hoc thm kh bng giy thm. Khi tiu bn kh hn em quan stdi knh hin vi.2. Phng php nhum Gram:y l phng php thng c dng trong thc nghim vi sinh vt hc.Phng php nhum kp do nh bc hc ngi an mch Christian Gram vnhng ngi cng tc a ra nm 1884. Vi cch nhum ny ta cth phn bit vi sinh vt thnh 2 nhm: Gram dng (G+) v Gram m (G-).Vi sinh vt bt mu xanh tm sau khi nhum gi l vi sinh vt Gram dng (baogm hu ht x khun, trc khun sinh bo t hiu kh, nm mc, nm men, cukhun). Cn nhng vi sinh vt bt mu hng gi l Gram m nh trc khun sinhbo t k kh, mt s cu khun nh lu cu khun, no m cu khun,cc vi khun ng rut)a. Nguyn tc: Da trn kh nng bt mu ca t bo cht v mng t bovi thuc nhum tm kt tinh v iot m hnh thnh nn hai loi phc cht khcnhau:- Loi phc cht th nht vn gi nguyn mu ca thuc nhum nn khngb ra tri khi x l bng cn. Vi sinh vt c phc cht ny thuc loi gram dng.- Loi phc cht th hai khng gi c mu ca thuc nhum nn mt mukhi b x l bng cn v bt mu ca thuc nhum b sung. Vi sinh vt c phccht ny thuc loi gram m.b. Cch tin hnh:Vt bi

Ph tm kt tinh ( Crystal violet )15sMu nhumRa ncRa ncTy cnRa ncPh lugolRa nc

1pht

1pht

15s

15s

15s

Ph safranin hoc Fuchsin

1pht

15s

D. Kt qu th nghim quan st vi khun bng phng php nhum Gram i vi 3 chng vi khun mu: Ecoli,Bacillus,Pseudomonas.a. Chng E.coli bt mu Gram m.

b. Chng Bacillus bt mu Gram dng (khng r lm do vi khun cn non)

c. Chng Pseudomonas bt mu Gram m:

Bi 3 : PHN NG SINH HAMC TIU:Nm c mc ch kho st nhng tnh cht sinh ha ca vi khun.Trnh by c nguyn tc ca nhng phn ng sinh ha thng thng.Quan st cc phn ng sinh ha v nhn xt kt qu.THC HNH:Nhng phn ng lin h n ngun thc n c cha Nit.Phn ng Indol:Nguyn tc:Pht hin kh nng oxi ha tryptophan thnh cc dng ca indol: Indole, Skatole (methyl indole) v indole-acetate.Mt s vi khun nh Ecoli c enzyme Tryptophanase chuyn Tryptophan thnh Indol.Mi trng:Tryptophan hoc peptone broth.Ta nui cy vi khun mi trng nc pepton khng cha ng trong 24-48h trong t 37oCThuc th:KovacssKt qu :

Dng tnh: xut hin vng mu trn b mt mi trng sau 5 pht. m tnh: mu vng (mu ca thuc th Kovacss)Kh nng to thnh H2S :Nguyn tc : Nhiu vi khun s dng nhng hp cht cha Sodium thiosunfuate th c th to thnh H2S.

Mi trng : Mi trng KIA gm glucose v lactose (c th ln men carbohydrat), phenol (ch th pH), pepton (ngun carbon/nitrogen) v mui st cng vi thiosulfate natri.Kt qu :Dng tnh khi lm cho mi trng thch thnh mu en.

Phn ng phn gii Ur :Nguyn tc:S dng pht hin kh nng phn ct urea thnh ammonia do hot tnh ca urease t vi sinh vt v kt qu l mi trng b kim ha do NH3 c to ra.

Mi trng: Urea broth (hoc Urea agar)Kt qu : Dng tnh: mi trng c mu hng m tnh: mi trng khng i mu (vng cam)

Nhng phn ng lin quan ti ngun thc n c Carbon :Kh nng ln men ng:Phn ng MR (methyl red)Mc ch: xc nh vi sinh vt sn xut v duy tr cc acid bn trong qu trnh ln men glucose.Mi trng MR-VPChng vi sinh vt

Kt qu, mu (Dng tnh), mu vng: (m tnh)

Methyl Red

24h, 37oC

Chng Vi Sinh Vt

Mi Trng

2-4 ngy

Dng tnh

m tnh

C ch:C s sinh ha:Di 4,45,0 5,8Trn 6,0

Cht ch th pH: methyl red

MR (+) cng ko di thi gian nui cy mi trng cng acidMR (-) cng ko di thi gian nui cy cc cht c tnh acid b chuyn ha mi trng dn trung tnh Thi gian 2 5 ngy 37oCC ch: mt s vi sinh vt c kh nng chuyn ha Glucose qua qu trnh ng phn to thnh acid pyruvic, ri t Acid pyruvic chuyn ha thnh acetyl coenzyme A, i qua chu trnh Kreps to ra cc acid: acid citric, acid succinic, acid fomic, a cid malic, a cid oxalic hoc t Acid pyruvic chuyn trc tip thnh cc acid lactic, acid fomic,Trong mi trng MR-VP c PH= 6.90.2 c K2HPO4 (m phosphat) gip n nh PH, khi c mt cc acid hu c mnh s tn cng v ph v th PH n nh lm cho PH gim mnh, PH