PLATING MEDIA FOR ROUTINE BACTERIOLOGY

(Bailey and Scott 12th ed and Mahon)

MEDIUM COMPONENTS PRIMARY PURPOSE
Alkaline Peptone
0.5-1.0% NaCl For Vibrio spp.
Water
Potato-glycerol-based medium with 15-20%
Bordet-Gengou
defibrinated blood. Contaminants inhibited by For Bordetella pertussis
agar
methicillin
Bismuth sulfite and brilliant green inhibits gram
Bismuth sulfite positive bacteria, most lactose-fermenting For isolation of Salmonella
agar (BSA) intestinal normal flora and Shigella spp.
Ferrous sulfate detects H2S production
Contains Brain and beef hear which provides For cultivation of non-
Brain Heart
nutrients; peptones, glucose, NaCl and buffers are fastidious and moderately
Infusion (BHI)
also added fastidious microorganisms
Yeast extract, agar, -ketoglutarate, and salts
Buffered charcoal- supplemented with L-cysteine enhance recovery
yeast extract of Legionella; activated charcoal absorbs toxic For Legionella spp.
(BCYE) agar compounds; ferric pyrophosphate which provides
iron; HCl and ACES buffer are also added
Casein medium Contains casein For Actinomycetes
Cetrimide (cetyltrimethyl ammonium
Selects Pseudomonas
Cetrimide agar bromide/CTAB)
aeruginosa in specimens
(pseudosel agar) Magnesium chloride and potassium sulfate
with mixed flora
stimulate pyocyanin production
Brucella Agar-based since it contains Na
bisulfite enhances recovery of microaerophiles by
lowering redox potential;
Contains 10% sheep blood plus antibiotics:
vancomycin to inhibit gram positive cocci,
Campy-blood agar trimethoprim to inhibit swarming strains of For Campylobacter spp.
Proteus,
polymyxin B to inhibit gram negative bacilli,
amphotericin B to inhibit fungi and yeasts,
cefoperazone (cephalothin is previously used)
inhibits Pseudomonas and Enterobacteriaceae
Cefoperazone, Blood-supplemented enrichment medium plus
Vancomycin, antibiotics
For Campylobacter spp.
Amphotericin Cefoperazone (inhibit growth of most gram-
(CVA) medium negative bacteria)
 Vancomycin (inhibits gram-positive bacteria)
Amphotericin (inhibits yeast)
For Yersinia spp.;
Cefsulodin- Peptone base with yeast extract, mannitol, and
Yersinia enterocolitica show
Irgasan- bile salts
bulls eye colonies;
Novobiocin (CIN) Antibiotics: Cefsulodin, Irgasan and Novobiocin
may be useful for isolation of
agar Indicators: Neutral red and Crystal violet
Aeromonas pp.
For fastidious organisms
Peptone base, enriched with solution of 2%
Chocolate agar such as Haemophilus and
hemoglobin or isovitalex
pathogenic Neisseria spp.
Columbia Colistin-
Columbia agar base with 10 mg colistin per liter,
Nalidixic acid For gram-positive cocci
15 mg nalidixic acid per liter, and 5% sheep blood
(CNA) agar
Cycloserine and cefoxitin inhibit the growth of
intestinal normal flora;
Fructose to check for fermentation; and neutral
Cycloserine-
red as pH indicator For Clostridium difficile
Cefoxitine-
VARIATION: uses mannitol instead of fructose and yellow colonies
Fructose Agar
bromthymol blue instead of neutral red; egg yolk
suspension can also be added to detect lecithinase
and lipase activity
Infusion agar with 5% sheep blood. Reduction of
Cystine-tellurite For C. diphtheriae (black
potassium tellurite by C.diphtheriae produces
blood agar colonies with brown halos)
black colonies
Sodium citrate and Sodium desoxycholate inhibits
most non pathogenic enteric bacilli and gram
positive organism;
Desoxycholate For isolation of enteric
Lactose to detect fermentation
Citrate Agar pathogens
Ferric ammonium citrate to detect H2S
production
SHOULD NOT BE AUTOCLAVED
For Isolation and
Eosin methylene
Peptone base with lactose and sucrose. Eosin and differentiation of lactose-
blue (EMB) agar
methylene blue as indicators fermenting and non-lactose
(Levine)
fermenting enteric bacilli
Fletcher Semi-
Rabbit serum and hemoglobin For Leptospira
solid medium
Peptone base broth with glucose and mannitol. Selective (enrichment)
Gram-negative
Sodium citrate and sodium desoxycholate act as medium for enteric
broth (GN)
inhib pathogens
 itory agents
Differential, selective
Peptone base agar with bile salts, lactose, sucrose
medium for isolation and
Hektoen enteric and salicin, and ferric ammonium citrate.
differentiation of Salmonella
agar (HEA) Indicators include bromthymol blue and acid
and Shigella spp. from other
fuchsin
gram-negative enteric bacilli
Determines whether an
High carbohydrate and Low peptone content
Hugh-Leifson organism is oxidative,
Bromthymol blue as pH indicator
OF Medium fermentative or
(alk = blue; acid = yellow)
biochemically inert
Lactose and glucose (10:1 ratio);
Phenol red as pH indicator; For determination of glucose
Kliglers Iron Agar Peptone as carbon or nitrogen source; or lactose fermentation, gas
Sodium thiosulfate and ferric ammonium sulfate and H2S production
as sulfur source and H2S indicator;
known as Modified Todd-Hewitt broth;
Peptones, yeast extract, dextrose;
Lim Broth For Streptococcus agalactiae
Colistin and Nalidixic acid inhibits gram negative
organisms
Enhances metachromatic
Loeffler High serum content and eggs to detect proteolytic
granule formation of
Coagulated Serum activity
Corynebacterium
Slant
diphtheriae
Potato flour, egg and glycerol as nutrient source
and as detoxifying agent;
Asparagine for maximum production of niacin;
Malachite green inhibits growth of other bacteria
medium should be kept away from light since
Lowenstein-Jensen
malachite green is light sensitive; For Mycobacterium spp.
Medium
GRUFT MODIFICATION: addition of penicillin and
nalidixic acid to make it selective, and RNA to
increase rate of isolation
PETRAN & VERA MODIFICATION: cyclohexamide,
lincomycin and nalidixic acid are added
Peptone base with lactose. Isolation and differentiation
Gram-positive organisms inhibited by crystal of lactose-fermenting and
MacConkey agar
violet and bile salts. non-lactose fermenting
Neutral red as indicator enteric bacilli
Peptone base, mannitol, and phenol red indicator. Selective isolation of
Mannitol salt agar
Salt concentration of 7.5% inhibits most bacteria staphylococci
 Selective isolation of gram-
Phenyl ethyl Nutrient agar base. Phenyl ethanol inhibits
positive cocci and anaerobic
alcohol (PEA) agar growth of gram-negative organisms
gram-negative bacilli
Charcoal agar supplemented with horse blood,
Regan Lowe For Bordetella pertussis
cephalexin, and amphotericin B
Peptone base with lactose, ferric citrate, and
Salmonella- For Salmonella and Shigella
sodium citrate. Neutral red as indicator; inhibition
Shigella (SS) agar spp.
of coliforms by brilliant green and bile salts
Peptone and soy protein base agar with yeast Nonselective medium for the
Schaedler agar extract, dextrose, and buffers. Addition of hemin, recovery of anaerobes and
L-cystine, and 5% blood enriches for anaerobes aerobes
Peptone base broth. Sodium selenite toxic for
Selenite broth For Salmonella spp.
most Enterobacteriaceae
Peptone and soy protein base agar with lysed
horse blood. Vancomycin inhibits gram-positive
Skirrow agar For Campylobacter spp.
organisms; polymyxin B and trimethoprim inhibit
most gram-negative organisms
Streptococcal
Contains crystal violet, colistin, and trimethoprim For S. pyogenes and S.
selective agar
sulfamethoxazole in 5% sheep blood agar base agalactiae
(SSA)
Peptone base broth. Bile salts and sodium
Tetrathionate For Salmonella and Shigella
thiosulfate inhibit gram-positive organisms and
broth spp.
Enterobacteriaceae
Pancreatic digest of casein, soy broth, glucose Supports growth of
Thioglycollate enrich growth of most microorganisms; anaerobes, aerobes,
broth Thioglycollate, cystine and sodium sulfite act as microaerophilic and
reducing agents; fastidious microorganisms
Potassium tellurite For Corynebacterium
Tinsdale Agar Cystine diphtheriae (black colonies
Thiosulfate with brown halo)
Peptone base agar with yeast extract, sucrose (to
detect fermentation);
ferric citrate and sodium thiosulfate to detect H2S
Thiosulfate
production;
citrate-bile salt
Na citrate, bile salts (10% oxgall) and Na For Vibrio sp/
sucrose (TCBS)
thiosulfate inhibit gram positive bacteria and
agar
gram negative rods;
Bromthymol blue acts as indicator (alk = green; ac
= yellow)
 An enrichment broth for streptococci,
Todd-Hewitt broth
supplemented with nalidixic acid and gentamicin For S. agalactiae in female
supplemented
or colistin for greater selectivity thioglycollate and genital specimens
with antibiotics
agar reduce redox potential
Enrichment broth used for
All-purpose enrichment broth that can support
Trypticase soy subculturing various
the growth of many fastidious and non-fastidious
broth bacteria from primary agar
bacteria
plates
Yeast extract agar with lysine, xylose, lactose,
Isolation and differentiation
Xylose lysine sucrose, and ferric ammonium citrate;
of Salmonella and Shigella
desoxycholate Sodium desoxycholate inhibits gram-positive
spp. from other gram-
(XLD) agar organisms
negative enteric bacilli
phenol red as indicator