Quality Control of Culture media

Dr Mohammad RahbarIranian Reference Health Laboratory

Anaerobic sheep blood and laked blood agar

• Incubation Atmosphere Anaerobic• Length, and Temperature• 24-48 h, 35 °C• Control Organisms(ATCC ) Expected Reults• B. fragilis (25285) :Growth• C. perfringens (13124) : Growth beta

hemolysis• F. nucleatum (25586) : Growth• P. anaerobius (27337): Growth• P. melaninogenica (25845) : Growth

Blood agar—nonselective sheep blood agarmedia

• Incubation Atmosphere,• Aerobic or CO2,• Length, and Temperature; 18-24 h , 35 °C• Control Organisms (ATCC® No.) Expected Results

• S. pyogenes (19615) : Growth beta hemolysis• S. pneumoniae (6305) : Growth alpha hemolysis• S. aureus (25923) : growth• E. coli (25922) : Growth

Blood agar-CAMP test (trypticase soy agar[TSA] with sheep blood only)

• Incubation Atmosphere :Anaerobi• Length, and Temperature :24-48 h, 35 °C• Control Organisms ATCC• S. aureus (33862) or (25923):Growth• S. agalactiae (12386) :Positive reaction

(Arrowhead area of clearing)• S. pyogenes (19615) :Negative reaction (No

arrowhead formation)

Blood agar—Selective sheep blood agar media

(Columbia [CNA] agar,

• Incubation Atmosphere : CO2°• Length, and Temperature :24-48 h, 35 °C• Control Organisms : Expected Results• S. pyogenes (19615) : Growth mbetahemolyric• S. pneumoniae (6305) : Growth alpha

hemolytic• S. aureus (25923) Growth• P. mirabilis (12453) : Growth

Blood agar—Selective sheep blood agar media

(Columbia [CNA] agar,

• Incubation Atmosphere : CO2°• Length, and Temperature :24-48 h, 35 °C• Control Organisms : Expected Results• S. pyogenes (19615) : Growth mbetahemolyric• S. aureus (25923) Growth• P. mirabilis (12453): Inhibition (partial)

Blood culture media.(TSI and BHI)

• Incubation Atmosphere : Anaerobic and aerobic

• Length, and Temperature : 5 days, 35 °C• Control Organisms : Expected Results• B. fragilis (25285): Growth• S. pneumoniae (6305) : Growth

Campylobacter agar (user quality control

required)

• Incubation Atmosphere :Reduced O2 with CO2,

• Length, and Temperature : 48 h, 42 °C• Control Organisms : Expected Results

• C. jejuni (33291) : Growth• E. coli (25922): Inhibition (partial)

Chocolate agar

• Incubation Atmosphere : CO2• Length, and Temperature :24-48 h, 35 °C• Control Organisms ATCC• N. gonorrhoeae (43069) : Growth• H. influenzae (10211) : gROWTH

Cefsulodin irgasan novobiocin (CIN)agar

• Incubation Atmosphere : Aerobic• Length, and Temperature : 24-48 h, 25 oC• Control Organisms ATCC:• Y. enterocolitica (9610) :Growth; deep red center transparent

border (bull’seye)

• E. coli (25922) :Inhibition (partial to complete)

• P. aeruginosa (27853) Inhibition (partial to complete)

• E. faecalis (29212) Inhibition (partial to complete)

•

Buffered charcoal yeast extract(BCYE) (CYE/BCYE) agar

• Incubation Atmosphere,:Aerobic• Length, and Temperature :48-72 h, 35 °C• Control Organisms• L. pneumophila (33152) :Growth; yellow-green

fluorescence under long-wave• UV light• L. bozemanii (33217) :Growth; blue-white

fluorescence• L. micdadei (33204 :Growth

Enrichment broths for enterics(gram-negative [GN] broth, selenite

broths)

• Incubation Atmosphere,:Aerobic• Length, and Temperature :18-24h, 35 °C• Control Organisms• S. typhimurium (14028) : Growth on subculture• S. sonnei (9290) : Growth on subculture (may be

inhibited by selenite• media)• E. coli (25922): Inhibition (partial to complete) on

subculture Growth on subculture from GN broth

Eosin methylene blue media(Levine EMB agar; EMB agar,

modified)

• Incubation Atmosphere,: Aerobic• Length, and Temperature : 18-24 h, 35 °C• Control Organisms:• S. typhimurium (14028) : Growth, colorless to

amber colonies• E. coli (25922): Growth, blue-black colonies with

green metallic• sheen• E. faecalis (29212): Inhibition (partial)

Hektoen enteric (HEK) agar

• Incubation Atmosphere, :Aerobic• Length, and Temperature :18-24 h, 35 °C• Control Organism:• S. typhimurium (14028): Growth, colonies blue to

green-blue with black• centers• S. flexneri (12022) :Growth, colonies green to blue-

green• E. faecalis (29212):Inhibition (partial); colonies yellow• E. coli (25922) :Inhibition (partial to complete);

colonies yellow to salmon colored

MacConkey agar

• Incubation Atmosphere, :Aerobic• Length, and Temperature :18-24 h, 35 °C• Control Organism

E. coli (25922) : Growth, pink colonies• P. mirabilis (12453) : Growth, colorless colonies,

partial inhibition of swarming• S. typhimurium (14028): Growth, colorless

colonies• E. faecalis (29212) : Inhibition (partial)

Mannitol salt agar

• Incubation Atmosphere, :Aerobic• Length, and Temperature :24-48h, 35 °C• Control Organism;• S. aureus (25923) : Growth, colonies have

yellow zones at 48 h• S. epidermidis (12228): Growth, colonies have

red zones at 48 h• P. mirabilis (12453) : Inhibition (partial)

Salmonella-Shigella (SS) agar

• Incubation Atmosphere, :Aerobic• Length, and Temperature :24 h, 35 °C• Control Organism• S. typhimurium (14028) : Growth, colonies

colorless with or without black centers• S. flexneri (12022) : Growth, colorless colonies• E. faecalis (29212) : Inhibition (complete)• E. coli (25922) : Inhibition (partial to complete;

colonies pink to rose-red with precipitate)

Selective media for pathogenic Neisseria spp.(TM)

• Incubation Atmosphere, :Co2• Length, and Temperature :24 -48h, 35 °C• Control Organism:• N. gonorrhoeae (43069) :Growth• N. meningitidis (13090) :Growth• P. mirabilis (43071) :Inhibition (partial) use only for media• containing trimethoprim• E. coli (25922) :Inhibition (partial)• N. sicca (9913) :Inhibition (partial)• C. albicans (60193) :Inhibition (partial)• S. epidermidis (12228) :Inhibition (partial)

Selective media for enterococci, with azide

• Incubation Atmosphere, :Aerobic• Length, and Temperature :24 -48h, 35 °C• Control Organism:• E. faecalis (29212) :Growth, blackening around

colonies• S. pyogenes (19615):Inhibition (partial to

complete)• E. coli (25922):Inhibition (partial) - Colorless

colonies on bile esculin agar

Selective media for enterococci, without azide

• Incubation Atmosphere, :Aerobic• Length, and Temperature :24 -48h, 35 °C• Control Organism:• E. faecalis (29212) :Growth, blackening

around colonies• S. pyogenes (19615):Inhibition (partial to

complete)

Thioglycolate broth, with or without indicator

• Incubation Atmosphere, :Aerobic, 48 h (tightened , cap

• Length, and Temperature : 48h, 35 °C• Control Organism:• B. fragilis (25285) : Growth• S. aureus (25923): Growth

Thioglycolate broth, enriched with vitamin Kand hemin

• Incubation Atmosphere, :Aerobic, 48 h (tightened , cap

• Length, and Temperature : 48h, 35 °C• Control Organism:• P. anaerobius (27337) : Growth• B. vulgatus (8482): Growth• C. perfringens (13124) : Growth

Tubed media (brain heart infusion and tryptic

soy broth)

• Incubation Atmosphere, :Aerobic• Length, and Temperature :18-24, 35 °C• Control Organism:• E. coli (25922) : Growth• S. aureus (25923) : Growth

Xylose lysine desoxycholate (XLD) agar

• Incubation Atmosphere, :Aerobic• Length, and Temperature : 24, 35 °C• Control Organism:• S. typhimurium (14028) :Growth - colonies red

with black centers• S. flexneri (12022) :Growth - colonies red• E. faecalis (29212) :Inhibition partial• E. coli (25922) :Inhibition (partial to complete;

colonies yellow to yellow-red)