Culture MethodsCulture MethodsII MBBSDr Ekta ChourasiaLecturer, Microbiology05.10.08 Dr Ekta, MicrobiologyCultureMethodsCultureMethodsIndications for culture -Isolate bacteria in pure cultures.Deonstrate their properties.!btain sufficient gro"th for preparation of antigens # for other tests.$yping bacterial isolates.%ntibiotic sensiti&ity.Estiate &iable counts.Maintain stock cultures.05.10.08 Dr Ekta, Microbiology$ypes of culture ethods$ypes of culture ethodsStreak culture or surface platingLa"n or carpet cultureStroke cultureStab culture'our plate ethod%naerobic ethods of culturing bacteria05.10.08 Dr Ekta, Microbiology%septic techni(ue%septic techni(ue05.10.08 Dr Ekta, MicrobiologyStreak CultureStreak Culture)outinely eployed for isolation'latinu * +ichroe loopshttp://vudat.msu.edu/ileadmi!/user"upload/vudat/lash/streakplate.s#05.10.08 Dr Ekta, Microbiology05.10.08 Dr Ekta, MicrobiologyLa"n or Carpet CultureLa"n or Carpet Culture,nifor surface gro"thBacteriophage typing%ntibiotic sensiti&ity testing'reparation of bacterial antigens # &accines05.10.08 Dr Ekta, MicrobiologyStroke CultureStroke Culture$ubes containing agar slopes-or slide agglutination # other diagnostic tests.05.10.08 Dr Ekta, MicrobiologyStab CultureStab CultureBy puncturing a suitable ediu "ith a long, straight charged "ire.-or gelatin li(uefaction, stock cultures # otility05.10.08 Dr Ekta, Microbiology'our 'late Method'our 'late Method. l of appropriately diluted inoculu is added to ./ l of olten agar and poured on petridish.Colonies appear through out the depth of ediu.,sed to estiate &iable count, recoended ethod for (uantitati&e urine cultures.05.10.08 Dr Ekta, MicrobiologyBroth CultureBroth CultureInoculated by a charged loop, pipette or syringes.-or blood cultures # sterility testing.05.10.08 Dr Ekta, Microbiology%naerobic Culture Methods%naerobic Culture Methods%naerobic condition can be achie&ed by.. displaceent of !0 "ith other gases.0. absorption of !0 by cheical or biological eans1. reduction of !0

05.10.08 Dr Ekta, Microbiology.. Displaceent Method.. Displaceent MethodDisplaceent of !0 "ith gases like 20 , +0 , 2e or C!0 .)arely produces coplete anaerobiosis. e.g. Candle 3ar 05.10.08 Dr Ekta, Microbiology0. Cheical or Biological Method0. Cheical or Biological Method%lkaline pyrogallol 4 pyrogallic acid in +a!25 absorbs !0.6ello" phosphorous.)osenthal ethod - Mi7ture of chroiu # sulphuric acid.McIntosh - -ildes8 9ar.:aspak05.10.08 Dr Ekta, MicrobiologyMcIntosh - -ildes8 9arMcIntosh - -ildes8 9arMost reliable # "idely used anaerobic ethodCoplete anaerobiosisCatalyst ; palladinised asbestos05.10.08 Dr Ekta, Microbiology:aspak:aspakMethod of choice for preparing anaerobic 3ars.Coercially a&ailable as disposable en&elope, containing cheicals "hich generate 20 , C!0 "ith the addition of "ater.)educed ethylene blue is used as indicator. )eains colorless anaerobically but turns blue on e7posure to !0 05.10.08 Dr Ekta, Microbiology1. )eduction of !7ygen1. )eduction of !7ygen,sing reducing agents like .< glucose, =..< thioglycollate, =..< ascorbic acid.)obertson8s Cooked Meat Mediuost "idely used fluid ediu-at free inced cooked eat in broth, "ith a layer of sterile &aseline o&er it.05.10.08 Dr Ekta, Microbiology%utoated Methods%utoated MethodsBactec - blood culture ethod; $he saple to be tested is inoculated into one or ore &ials "hich are inserted into the BACTEC fluorescent series instruent for incubation and periodic reading. ; Each &ial contains a cheical sensor "hich can detect increases in C!0 produced by the gro"th of icroorganiss. ; $he sensor is onitored by the instruent e&ery ten inutes for an increase in its fluorescence, "hich is proportional to the aount of C!0 present. ; % positi&e reading indicates the presupti&e presence of &iable icroorganiss in the &ial.