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MLT-Culture Media

Microbiology

QuestionAnswer
The purpose of culture media? Growth Isolation Identification Enumerating viable cells Keeping culture
Growth requirements in Bacteria? Water: 80% Dry weigth: 20% protein, lipid, carbohydrate
Usual components of culture media? Water Agar for support Peptone to provide nitrogen Meat extracts to provide aa, vitamins Yeast extracts to provide nucleotides Mineral salts Carbohydrates: carbon and energy sources
Culture media can be in the forms of? 1.Solid(1.2% agar): for observing colonies 2.Semi-solid(0.2% agar): for observing motility, or biochemical reactions 3.Liquid: for bacterial count
Categories of culture media? Basic media Enriched media Differential media Selective media Enrichment media Transport media
What is basic media? Simple media, e.g. nutrient agar, nutrient broth
What is enriched media? With additional nutrients to support growth of pathogens, eg: Blood, serum, extra peptones, vitamins etc.
What is blood agar? Agar base (Columbia agar) + blood (5-7%) Supports growth of most organisms Differentiates haemolysis Made selective by adding antibiotics
What is chocolate agar? Agar base Columbia agar base Heated blood More nutritious than blood agar For e.g. H. influenzae
What is selective media? Contains inhibitory substances prevents or slows down growth of unwanted bacterial species. For isolation of target bacterial species from non sterile specimens
Example of selective media? Deoxycholate citrate agar (DCA) isolate Salmonella species from faeces. MacConkey agar (Mac) isolate Gram negative bacterial species from wounds etc.
What is differential media? Contains carbohydrate, amino acid or other substrates, indicators or dyes that differentiate bacterial species.
Example of differential media? E.g. DCA, MacConkey Contains lactose & neutral red (pH indicator) Differentiate lactose fermenting from non lactose fermenting bacteria
What is MacConkey agar? Selective and differential Selective agent: bile salt & crystal violet Isolate Gram negative bacterial species Differential agents: lactose and neutral red Supports growth of all Enterobacteriaceae Useful for stool, wound swab & pus
What is Deoxycholate citrate agar (DCA)? Selective & differential for isolation of Salmonella & Shigella Selective agents: Deoxycholate inhibits growth of GP organisms,Sodium citrate toxic for intestinal organisms Differential agents:Lactose & neutral red, Sodium thiosulphate & Ferric citrate
What is Xylose lysine deoxycholate (XLD) agar? For isolation of Salmonella & Shigella Selective agents: deoxycholate inhibits growth of GP organisms Differential agents:Xylose, lactose, sucrose & phenol red;Lysine;Sodium thiosulphate & ferric ammonium sulphate
What is TCBS (Thiosulphate citrate bile salt sucrose agar)? Selective & differential For isolation of Vibrio species Selective agents: Thiosulphate , citrate, bile salt Differential agents:Sucrose & bromo thymol blue
What is Cysteine Lactose Electrolyte Deficient (CLED) agar? Differential:Lactose & bromo thymol blue Isolation and differentiation of urinary microbes
What is enrichment media? Usually fluid medium Contains enrichment factor or inhibitory substances E.g. selenite F broth, alkaline peptone water
What is Alkaline peptone water? Enrichment medium for Vibrio species Alkaline pH (pH 8.4) Subculture after 6 hours
What is Robertson's cooked meat broth? Meat contains reducing substances anaerobic
What is Transport media? Ensure viability of aerobic and anaerobic pathogens Stuart Transport medium, Amies Transport medium, Cary Blair Transport medium
Difference between blood agar and chocolate agar? blood agar: X factor CHO agar: X and V factor Haemophilus grows on CHO only
Principle of MacConkey agar? lactose fermenter: breakdown lactose and produce acid, pH indicator turns red at acidic pH Non lactose fermenter: cannot breakdown lactose and use protein, alkaline amines are produced, pH indicator remains no change or yellow
Example of XLD agar? E.coli: yellow colony Shigella: colorless/red colony Salmonella: black centre colony
Purpose of TCBS agar? Differentiate between different Vibrio species V. cholerae (yellow) V. parahaemolyticus (pale crystal green)
Created by: kencho
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