8/14/2019 Culture Media Note

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CULTURE MEDIA

Peptone water(peptone+NaCl+dist. Water)

Basal media

Autoclave at 110C for 10minutes

Base for all sugar media for non-fastidious bacteria.

Nutrient broth( peptone water+NaCl+meat

extract)

Base for most media used fororganisms growth.

Nutrient agar plate(broth + 2% agar)

Nutrient agar slope

Allows separate colonyformation

Colonial morphologyidentification

Quantitation: no. & relativeproportion of different bacterialspecies originally present inspecimen can be estimated.

Pure cultures can be obtained

Deep agar tube

Basal mediaAutoclave at 110C for 10

minutes

to test oxygen requirement ofbacteria

Soft agar tube(in small tube)

*both with cotton-woolstoppers.

For keeping stock cultures by stabinoculation.

To demonstrate motility oforganism.

Cooked meat medium(Robertsons medium)

Reducing agent:haem, unsaturatedfatty acid.

For growth of anaerobic bacteria:

Clostridium botulinum,clostridium tetani, Anaerobicstreptococci.

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8/14/2019 Culture Media Note

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Paranitro benzoic acid(PBNA)Medium

Blood agar(nutrient agar+ steriledefibrinated blood)

Enriched media

Autoclave at 120 for 20minutes

Growth of atypical mycobacteria.

Growing delicate or fastidiousbacteria

Differential medium:differentiating org. According to

their haemolytic action on blood.

Chocolote agar(nutrient agar+ steriledefibrinated blood+ heated to80C for 10 min)

Enriched mediaAutoclave at 120 for 20

minutes

Culture ofHaemophilus &Neisseria

Loefflers Serum Slope(3 pt sterile serum+ 1 ptglucose broth)

Screw capped bottles or tubes

Enriched mediaSolidified by inspissations

( 70-80C for 2 hrs)

Prepared under sterileconditions

Morphology ofC. Diphtheria

Egg Saline Medium(2.5 pt egg yolk+ 1 pt saline)

Screw-capped bottles

Enriched MediaSolidified by inspissations

( 70-80C for 2 hrs)Prepared under sterile

conditions

Culture of tubercle bacilli

Selenite F broth

Tetrathionate broth

Enrichment media Favour growth of typhoid &

parathyphoid organisms.(shigella, salmonella)

Do not favours growth of E. Coli

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8/14/2019 Culture Media Note

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Mac Conkeys medium(peptone water+agar+bile

salt+lactose+neutralred(phenol red).

Selective media

Steaming for 30-60 min(autoclave)

Sodium taurocholate(bilesalt)

*selective agent or inhibitorysubstance:

- growth of gram +ve bacteria

Sugar lactose-fermented byE. Coli

Indicator-phenol red

*pinkfermenting colonies

*yellownon-fermenting colonies

To grow gram vebacteria.

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 ofvibrio cholera.

Colonies appear yellow

against green medium.

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8/14/2019 Culture Media Note

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Mannitol Salt Agar

Indicatorphenol red Sugarmannitol

Selective media

NaClinhibitory substance,allows growth ofstaphylococci, inhibits otherorganisms.

Mannitol fermentation of

staphylococcicoloniessurrounded by yellowhaloes.

Lowensteins JensenMedium

(beatean eggs, mineralsalts, malachite green)

Solidified byinspissation

Selective media

Sel. Agentmalachitegreen- Growth of other bacteria

other than TB.

Glycerolenhance growth ofhuman type.

Pyruvateenhance growth ofbovine type.

Cultivation oftuberclebacilli

Thayer Martin medium Selective medium

Colistin, nystatin,vancomycin

For isolation ofNeisseria

Blood tellurite medium(Mcleods medium)

(blood agar+ potassiumtellurite)

Selective medium:

K tellurite

Allows growth ofC.Diphtheria as black

colonies.

Sugar media( peptonewater, 0.5-1.0% sugar:)

Glucose

Sugar media

Indicatorsphenol red,

Placed in tubes containing:

inverted Durhams tubes

to trap air bubbles in case

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8/14/2019 Culture Media Note

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Maltose

Mannitol

Lactose

Sucrose

Change their color w changeof pH

Yellowacid pH

Redalkaline pH

Tyndallisation for 30 min for3 successive days.

of fermentation with gas.

An indicator media todifferentiate bacteriaaccording to fermentation ofsugars.

Transport medium: Stuarts transport

medium

Transport medium Semi-solid

Non-nutrient agar withthioglycollic acid(reducingagent)+ electrolytes

Preservation of delicatepathogens during transit tolab.

Originally for NeisseriaGonorrhea.

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