CULTURE MEDIA

Peptone water (peptone+NaCl+dist. Water) Nutrient broth ( peptone water+NaCl+meat extract) Nutrient agar plate (broth + 2% agar) Nutrient agar slope Basal media Autoclave at 110C for 10 minutes Base for all sugar media for nonfastidious bacteria. Base for most media used for organisms growth.

Allows separate colony formation Colonial morphology identification Quantitation: no. & relative proportion of different bacterial species originally present in specimen can be estimated. Pure cultures can be obtained

Deep agar tube Soft agar tube (in small tube) *both with cotton-wool stoppers. Basal media Autoclave at 110C for 10 minutes

to test oxygen requirement of bacteria

For keeping stock cultures by stab inoculation. To demonstrate motility of organism. For growth of anaerobic bacteria: Clostridium botulinum, clostridium tetani, Anaerobic streptococci.

Cooked meat medium (Robertsons medium)

Reducing agent: haem, unsaturated fatty acid.

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Paranitro benzoic acid(PBNA) Medium Enriched media Blood agar (nutrient agar+ sterile defibrinated blood) Chocolote agar (nutrient agar+ sterile defibrinated blood+ heated to 80C for 10 min) Loefflers Serum Slope (3 pt sterile serum+ 1 pt glucose broth) Screw capped bottles or tubes

Growth of atypical mycobacteria. Growing delicate or fastidious bacteria Differential medium: differentiating org. According to their haemolytic action on blood. Culture of Haemophilus & Neisseria

Autoclave at 120 for 20 minutes

Enriched media Autoclave at 120 for 20 minutes Enriched media Solidified by inspissations ( 70-80C for 2 hrs) Prepared under sterile conditions Enriched Media Solidified by inspissations ( 70-80C for 2 hrs) Prepared under sterile conditions Enrichment media

Morphology of C. Diphtheria

Egg Saline Medium (2.5 pt egg yolk+ 1 pt saline) Screw-capped bottles

Culture of tubercle bacilli

Selenite F broth Tetrathionate broth

Favour growth of typhoid & parathyphoid organisms. (shigella, salmonella) Do not favours growth of E. Coli

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 Mac Conkeys medium (peptone water+agar+bile salt+lactose+neutral red(phenol red).

Selective media

To grow gram ve bacteria.

Steaming for 30-60 min (autoclave)

Sodium taurocholate(bile salt) *selective agent or inhibitory substance: - growth of gram +ve bacteria

Sugar lactose-fermented by E. Coli

Indicator-phenol red *pinkfermenting colonies *yellownon-fermenting colonies TCBS Agar (sodium thiosulphate, sod. Citrate, bile salt, sucrose) Selective media

Indicatorbromothymol blue pH8.0-8.6(alkaline) Steaming for 30-60 min (autoclave)

For selective isolation of vibrio cholera. Colonies appear yellow against green medium.

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Mannitol Salt Agar Indicatorphenol red Sugarmannitol

Selective media

NaClinhibitory substance, allows growth of staphylococci, inhibits other organisms.

Mannitol fermentation of staphylococcicolonies surrounded by yellow haloes.

Lowensteins Jensen Medium

(beatean eggs, mineral salts, malachite green)

Solidified by inspissation Thayer Martin medium

Selective media Sel. Agent malachite green - Growth of other bacteria other than TB. Glycerolenhance growth of human type. Pyruvateenhance growth of bovine type.

Cultivation of tubercle bacilli

For isolation of Neisseria

Selective medium Colistin, nystatin, vancomycin

Blood tellurite medium (Mcleods medium) (blood agar+ potassium

tellurite)

Selective medium: K tellurite Sugar media

Allows growth of C. Diphtheria as black colonies.

Sugar media( peptone

water, 0.5-1.0% sugar:)

Glucose

Indicatorsphenol red,

Placed in tubes containing: inverted Durhams tubes to trap air bubbles in case

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Maltose Mannitol Lactose Sucrose

Change their color w change of pH Yellowacid pH Redalkaline pH Tyndallisation for 30 min for 3 successive days. Transport medium Semi-solid Non-nutrient agar with thioglycollic acid(reducing agent) + electrolytes

of fermentation with gas. An indicator media to differentiate bacteria according to fermentation of sugars.

Transport medium: Stuarts transport medium

Preservation of delicate pathogens during transit to lab. Originally for Neisseria Gonorrhea.

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