1. Quality Control of Culture media
Dr Mohammad Rahbar
Iranian Reference Health Laboratory

2. 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

3. Blood agar—nonselective sheep blood agar media
Incubation Atmosphere,
Aerobic or CO2,
Length, and Temperature; 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

4. 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)

5. 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

6. 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)

7. 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

8. 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)

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

10. Cefsulodin irgasan novobiocin (CIN) agar
Incubation Atmosphere : Aerobic
Length, and Temperature : 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)

11. 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

12. Incubation Atmosphere,:Aerobic Length, and Temperature :18-24h, 35 °C
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

13. Eosin methylene blue media (Levine EMB agar; EMB agar, modified)
Incubation Atmosphere,: Aerobic
Length, and Temperature : 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)

14. 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

15. 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)

16. 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)

17. 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)

18. 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)

19. 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

20. 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)

21. 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

22. Thioglycolate broth, enriched with vitamin K and 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

23. 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

24. 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)