Know your culture – a quick reference guide

Culture is a technique where we grow microorganisms in a nutritional liquid or on a solid medium in order to increase numbers and simplify identification. It is also important as a first step in several laboratory techniques and for testing of antimicrobial sensitivity.

There are many types of medium available for culturing microorganisms, and so it is important to know which is best to use – especially for organisms that are difficult to grow.

Culture table

Many of these media are available commercially, but for some it can be more cost effective to prepare the media yourself. For completeness, recipes for all media above are listed in alphabetical order. Adjustment of these recipes may be necessary for some cultures, but these will provide a good starting point. Most of the recipes include ‘agar’, which can be left out to make liquid cultures, but some rely on the use of nutrient agars which will require replacing with another nutrient source to make a liquid medium for culture.

 


 

Contents:

 


 

Bacteroides Bile-Esculin

Bacteroides bile-esculin with gentamycin and bile

Bacteroides bile-esculin (with gentamicin and bile) agar:

  • 45g/l Tryptic Soy Agar
  • 20g/l Ox bile
  • 1 g/l Esculin
  • 5g/l Ferric Ammonium Citrate
  • 12mg/l Hemin
  • 100mg/l Gentamicin

Bordet-Gengou agar with methicillin or cephalexin:

  • 15g/l Agar
  • 5g/l Potato Infusion
  • 5g/l Sodium Chloride
  • 3g/l Meat Peptone
  • 2g/l Casein Peptone
  • 10ml/l Glycerol
  • 15% Sheep’s blood
  • Methicillin or Cephalexin as appropriate to your sample

 

bordetella-pertussis-charcoal-agar-plate CROPPED

Buffered Charcoal Yeast Extract Agar

Buffered Charcoal Yeast Extract agar:

  • 17g/l Agar
  • 5g/l Yeast Extract
  • 10g/l ACES Buffer
  • 3g/l Activated Charcoal
  • 5g/l Potassium Hydroxide
  • 1g/l alpha-Ketoglutarate
  • 4g/l L-Cysteine
  • 25g/l Ferric Pyrophosphate

Campylobacter-selective agar (for example):

  • 5g/l Agar
  • 12g/l Pancreatic Digest of Casein
  • 5g/l Peptic Digest of Animal Tissue
  • 5g/l Sodium Chloride
  • 4g/l Activated Charcoal
  • 3g/ Yeast Extract
  • 3g/l Beef Extract
  • 1 g/l Corn Starch
  • 1g/l Cycloheximide
  • 32mg/l Hemin
  • 32mg/l Cefoperazone
  • 20mg/l Vancomycin

 

CIN_Agar_800X600

Cefsulodin-Irgasan-Novobiocin agar

Cefsulodin-Irgasan-Novobiocin agar:

  • 20g/l Mannitol
  • 17g/l Peptone
  • 5g/l Agar
  • 3g/l Proteose Peptone
  • 2g/l Yeast Extract
  • 2g/l Sodium Pyruvate
  • 1g/l Sodium Chloride
  • 5g/l Sodium Deoxycholate
  • 5g/l Sodium Cholate
  • 30mg/l Neutral Red
  • 10mg/l Magnesium Sulfate
  • 4mg/l Irgasan®
  • 4mg/l Cefsulodin
  • 5mg/l Novobiocin
  • 1mg/l Crystal Violet

Cystine Heart agar:

  • 15g/l Agar
  • 10g/l Beef Heart (Infusion from 500g)
  • 10g/l Proteose Peptone
  • 10g/l Dextrose
  • 5g/l Sodium Chloride
  • 1g/l L-Cystine

 

gram -ve bacilli blood agar.jpg

Cystine-Tellurite Blood agar

Cystine-Tellurite Blood agar:

  • 15g/l Agar
  • 13g/l Casein Peptone
  • 5g/l Sodium Chloride
  • 5g/l Yeast Extract
  • 2g/l Beef Heart (Infusion from 500g)
  • 44mg/l L-Cystine
  • 50ml/l 1% Potassium Tellurite
  • 5% Sheep’s Blood

Fletcher’s medium with rabbit serum:

  • 5g/l Agar
  • 5g/l Sodium Chloride
  • 3g/l Peptone
  • 2g/l Beef Extract
  • 80ml/l Rabbit Serum

Gram-Negative Enrichment Broth:

  • 10g/l Pancreatic Digest of Casein
  • 10g/l Peptic Digest of Animal Tissue
  • 5g/l Sodium Chloride
  • 5g/l Sodium Citrate
  • 4g/l Dipotassium Phosphate
  • 2g/l Mannitol
  • 5g/l Monopotassium Phosphate
  • 1g/l Dextrose
  • 5g/l Sodium Deoxycholate

 

Hektoen Agar CROPPED

Hektoen agar

Hektoen agar:

  • 5g/l Agar
  • 12g/l Peptic Digest of Animal Tissue
  • 12g/l Lactose
  • 12g/l Sucrose
  • 9g/l Bile Salts
  • 5g/l Sodium Chloride
  • 5g/l Sodium Thiosulfate
  • 3g/l Yeast Extract
  • 2g/l Salicin
  • 5g/l Ferric Ammonium Citrate
  • 1g/l Acid Fuchsin
  • 64mg/l Bromothymol Blue

Horse Blood-Charcoal agar:

  • 12g/l Agar
  • 10g/l Beef Extract
  • 10g/l Casein/Meat polypeptone
  • 10g/l Starch
  • 5g/l Sodium Chloride
  • 4g/l Charcoal
  • 1mg/l Nicotinic acid
  • 10% Horse Blood

Kanamycin-Vancomycin Laked Blood agar:

  • 20g/l Peptamin
  • 17g/l Agar
  • 5g/l Sodium Chloride
  • 2g/l Yeast Extract
  • 1g/l Dextrose
  • 1g/l Sodium Bisulfite
  • 5mg/l Hemin
  • 1mg/l Vitamin K
  • 40ml/l Vancomycin
  • 40ml/l Kanamycin
  • 50ml Laked Sheep’s Blood

Leptospira Medium with Bovine Serum Albumin (BSA)-Tween 80:

  • Base medium (900ml)
    • 11g/l Agar
    • 1g/l Disodium Phosphate
    • 1g/l Sodium Chloride
    • 3g/l Monopotassium Phosphate
    • 1ml/l 10% Glycerol
    • 1ml/l 25% Ammonium Chloride solution
    • 1ml/l 10% Sodium Pyruvate solution
    • 1ml/l 0.5% Thiamine solution
  • Albumin Supplement (100ml)
    • 100g/l BSA
    • 125ml/l 10% Tween-80 solution
    • 100ml/l 0.5% Iron Sulfate solution
    • 10ml/l Magnesium Chloride-Calcium Chloride solution (see below)
    • 10ml/l 0.02% Cyanocobalamin solution
    • 10ml/l 0.4% Zinc Sulfate solution
  • Magnesium Chloride-Calcium Chloride solution:
    • 15g/l Calcium Chloride Dihydrate
    • 15g/l Magnesium Chloride Hexahydrate

 

Loeffler's coagulated serum medium

Loeffler’s Coagulated Serum Medium

Löffler’s (also spelled ‘Loeffler’) Coagulated Serum Medium:

  • 5g/l Dextrose
  • 5g/l Gelatin Peptone
  • 5g/l Beef Extract
  • 25g/l Sodium Chloride
  • 750ml/l Bovine Serum

 

MacConkey_agar_with_LF_and_LF_colonies.jpg

MacConkey agar

MacConkey agar:

  • 17g/l Peptone
  • 5g/l Agar
  • 10g/l Lactose
  • 5g/l Sodium Chloride
  • 5g/l Bile Salts
  • 30mg/l Neutral Red
  • 1mg/l Crystal Violet

MacConkey-Sorbitol agar:

  • As above, but including 10g/l sorbitol instead of lactose

Modified Thayer-Martin agar:

  • 23g/l Peptic Digest of Animal Tissue
  • 20g/l Agar
  • 5g/l Sodium Chloride
  • 5g/l Dextrose
  • 1g/l Starch
  • 200ml/l Lysed Blood
  • 5mg/l Colistin Methane Sulphate
  • 3mg/l Vancomycin

New York City agar:

  • 20g/l Agar
  • 15g/l Proteose Peptone
  • 5g/l Glucose
  • 5g/l Sodium Chloride
  • 4g/l Dipotassium Hydrogen Phosphate
  • 1g/l Corn Starch
  • 1g/l Potassium Dihydrogen Phosphate
  • 100ml/l Horse Blood Cells
  • 60ml/l Horse Plasma
  • Yeast Autolysate Supplement (Available commercially, recipe below)
    • 10g/l Yeast Autolysate fractions
    • 1g/l Glucose
    • 15g/l Sodium Bicarbonate
  • NYC Supplement, also known as C.L.A.T. antibiotics (available commercially, recipe below)
    • 5mg/l Colustin Methane Sulphonate
    • 4mg/l Linomycin
    • 1mg/l Amphotericin B
    • 5mg/l Trimethoprim Lactate

Pseudomonas cepacia agar:

  • 15g/l Agar
  • 5g/l Sodium Pyruvate
  • 3g/l Dipotassium Phosphate
  • 1g/l Monopotassium Phosphate
  • 5g/l Bile Extract
  • 1g/l Enzymatic Digest of Animal Tissue
  • 1g/l Ammonium Sulfate
  • 2g/l Magnesium Sulfate
  • 1g/l Ticarcillin
  • 20mg/l Phenol Red
  • 10mg/l Ferrous Ammonium Sulphate
  • 1mg/l Crystal Violet
  • 300 000Units Polymyxin B

Regan-Lowe Cephalixin agar:

 

salmonella-shigella agar.png

SalmonellaShigella agar

SalmonellaShigella agar:

  • 5g/l Agar
  • 10g/l Lactose
  • 5g/l Bile Salts
  • 5g/l Sodium Citrate
  • 5g/l Sodium Thiosulfate
  • 5g/l Beef Extract
  • 5g/l Enzymatic Digest of Casein
  • 5g/l Enzymatic Digest of Animal Tissue
  • 1g/l Ferric Citrate
  • 25mg/l Neutral Red
  • 33mg/l Brilliant Green

Selenite Enrichment Broth:

  • 10g/l Sodium Phosphate
  • 5g/l Pancreatic Digest of Casein
  • 4g/l Lactose
  • 4g/l Sodium Selenite

Stuart Medium with rabbit serum:

  • 10g/l Sodium Glycerophosphate
  • 3g/l Agar
  • 1g/l Sodium Thioglycollate
  • 1g/l Calcium Chloride
  • 2mg/l Methylene Blue
  • 80ml/l Rabbit Serum

 

Thiosulfate-citrate-bile salts-Sucrose agar.png

Thiosulfate-Citrate-Bile salts-Sucrose agar

Thiosulfate-Citrate-Bile Salts-Sucrose agar:

  • 20g/l Sucrose
  • 15g/l Agar
  • 10g/l Dipeptone
  • 10g/l Sodium Citrate
  • 10g/l Sodium Thiosulfate
  • 10g/l Sodium Chloride
  • 5g/l Yeast Extract
  • 5g/l Ox Bile
  • 3g/l Sodium Cholate
  • 1g/l Ferric Citrate
  • 40mg/l Bromothymol Blue
  • 40mg/l Thymol Blue

Tinsdale agar:

  • 20g/l Protease Peptone
  • 15g/l Agar
  • 5g/l Yeast Extract
  • 5g/l Sodium Chloride
  • 425g/l Sodium Thiosulfate
  • 3445g/l Potassium Tellurite
  • 24g/l L-Cystine
  • 100ml Serum

 

xldsalmonella

Xylose-Lysine-Deoxycholate agar

Xylose-Lysine-Deoxycholate agar:

  • 5g/l Agar
  • 5g/l Lactose
  • 5g/l Sucrose
  • 8g/l Sodium Thiosulfate
  • 5g/l L-Lysine HCl
  • 5g/l Sodium Chloride
  • 3g/l Yeast Extract
  • 1g/l Sodium Deoxycholate
  • 8g/l Ferric Ammonium Citrate
  • 80mg/l Phenol Red

 

Robert G Millar (University of Warwick)

Society for Applied Microbiology



Categories: Feature Articles

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