microbiology practical revision

8/10/2019 Microbiology Practical Revision

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Microbiology practicalrevision Dr. TanmayMehta

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CULTURE MEDIA



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RED

L!!D A"ARRI"#T RED!$A%UE

MACC!&'E()* A"AR$I&' REDTRA&*$ARE&T



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loo+ agar



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M/A

LF dry LF mucoid LNF



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C,ocolateC#!C!LATE A"AR$!TA**IUM TELLURITE L!!DA"AR

$!TA**IUM TELLURITE L!!DA"AR

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"reen

TC *TC * -IT# (ELL!-C!L!&IE* !.C#!LERAE



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&0trient agar

Lemon yellow golden yellow white

Swarming - proteus



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ILE 1TAUR!C#!LATE2 R!T#ADULT 3 $EDIATRIC !TTLE



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RCM

Proteolytic sacharolytic control



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Dorset egg 3 L4



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Lo5er)s ser0m slope



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*DA



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&&&



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*ensitivity +isc



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-ilson blair



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#o6 to +escribe colony onsoli+ me+i0m 7

1. !i"e2. !ha#e $circ%lar/o&al'3. !%rface $(mooth/ro%gh'$(hiny/non(hiny'4. Margin( $reg%lar/irreg%lar'5. )on(i(tency $friable/b%t%ro%(/&i(ci*'6. !tr%ct%re $tran(#arent/tran(l%cent/o#a+%e'7. )olo%r8. !ateletti(m/(,arming9. Me*i%m change

$#igment/hemoly(i(/blackening'



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#o6 to +escribe colony onsoli+ me+i0m 7

T%rbi*ity-ran%lar *e#o(it(

!%rface gro,th #ellicleo*or



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I!C#EMICAL

REACTI!&*



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Catalase test

3% Hydrogen peroxide in TT

8# 8 ! 8 9 8# 8 ! : ! 8 1gas b0bble2

Catalase from bacteria i( an en"yme that *ecom#o(e(hy*rogen #ero i*e$ 20 2 ' into ,ater an* o ygen o(iti&e

!ta#hylococc%( a%re%(egati&e )ontrol!tre#tococc%( (#ecie(



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Coag0lase test

!ta#hylococcal coag%la(e i( a #rotein that ha(#rothrombin like acti&ity ,hich can con&ert;brinogen into ;brin . visible clot ,ill

re(%lt.o0n+ Coag0lase< l(o kno,n a( cl0mpingfactor ,hich i( attache* to bacterial cell ,all andnot present in cultural fltrates *etecte* by

sli+e test.ree Coag0lase< t i( a thrombin like (%b(tance#re(ent in c0lt0ral ;ltrate . t form( a clot byreacting ,ith coag%la(e reacting factor 1CR 2#re(ent in (er%m. t i( *etecte* by t0be test.



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I M E.coli



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In+ole test

Inoculate and incu ate !%

acteria that #o((e( the en"yme tryptop,anase are ca#able ofhy*roly"ing an* *eaminating tryptop,an ,ith the #ro*%ction ofin+ole #yr%&ic aci* an* ammonia.

- o(iti&e controlEsc,eric,ia coli

rote%( &%lgari()itrobacter *i&er(%(*,ar*(iella tar*a

egati&e control'lebsiella pne0moniae



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Met,yl re+ test

Inoculate " Incu ate the #$ &P roth with a pure culture

The # at ,hich methyl re* *etect( aci* i( con(i*erably lo,er thanthe # for other in*icator(. Th%( to #ro*%ce a colo%r change the te(t

organi(m m%(t #ro*%ce large +%antitie( of (trong aci* from thecarbohy*rate (%b(trate

- o(iti&e controlEsc,eric,ia coli

egati&e controlnterobacter aerogene(.



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oges proska0er test

Inoculate & Incubate the MR/VP broth with a pure culture -1 mle=pose t,eme+i0m toatmosp,eric! 8 >? to >@min.

#o(iti&enterobacter aerogene(.

- egati&eEsc,eric,ia coli

-l%co(e $yr0vic aci+ (ome bacteria metaboli(e to acetoin$acetyl methyl carbinol' n the #re(ence of atmospheric O 2 &

40% KOH acetoin i( con&erte* to *imethyl carbinol :

naphthol re* col%or



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Citrate 0tili ation test

;o(er


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!=i+ase test

Electron donor bacteria (

(e%*omona( aer%gino(aeromona( (#ecie(ei((eria (#ecie()am#ylobacter (#ecie(?ibrio (#ecie(




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Urease test

tuart!" urea broth and #hri"ten"en!" urea agar

;leb(iella (#ecie(rote%( (#ecie()itrobacter(#ecie(Morganella(#ecie(




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$,enylalanine +eaminase test1$$A test2

PP' agar

rote%( (#ecie(Morganella(#ecie(ro&i*encia(#ecie(




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T*I agar

Me*i%mgl%co(e $ .1@' lactose 1> 2 an+ s0crose 1> 2henol re* in*icator

ferro%( $iron' (%lfateno fermentation no change > Ae*/Ae* (ello6 color occ%r( *%e to fermentation ofcarbo,y+ratesf gas i( #ro*%ce* the agar may be lifte+ o thebottom of the te(t t%be an* the agar may al(obreak apart.# 8 * #ro*%ction from the metaboli(m of ferro%((%lfate. i( in*icate* by blackening of the me*i%m.



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Re+/yello6

f !&L( "LUC!*E i( fermente* then thebottom of the (lant $calle* the b0ttF2,ill t%rn yello6.

o,e&er there i( not enough glucose inthe me*i%m to turn the top of the slant

yellow . Chen the gl%co(e i( %(e* %# peptone $,hich i( al(o #re(ent in the me*i%m' i( be 0tili e+ $aerobically' forming an alkaline byGpro+0ct . !o the to# of the (lant $,hereo ygen i( #re(ent' may t%rn +eeper re+.



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(ello6/yello6

f -l%co(e an* acto(e or -l%co(e an* !%cro(e are fermente*

then the entire (lant ,ill t%rn yello,.



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Slant Butt Gas H 2S OrganismAlkaline

(Red) NR - - Pseudomonas

aeruginosa

Acid

(Yellow)

Acid

(Yellow)

+ - E. coli

Alkaline( Red)

Acid(Yellow)

+ Salmonella typhiCitrobacter freundiiProteus mirabilis

Alkaline( Red)

Acid(Yellow)

- - Shigella



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*pecimen



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Ro0n+ 6orm in bottle



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$arasitology stool



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Trop,o oite G giar+ia



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Cysts

E.#. "IARDIA



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E""

#.&A&A TAE&IA



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E. ermic0laris egg



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Instr0ments



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$etri+is,



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-ire loop



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Urine container



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*tool container



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#I TriG+ot



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Imm0nocomb



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Can+le ;lter



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$# strips



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microbiology practical revision

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