11/24/2009

Wound Cultures

Selection of Culture Media

Wound culture specimens are cultured for the isolation of common and potential pathogens

Wound Cultures Culture Setup

UNMC Clinical Laboratory Science Program
Linsey Donner, MT (ASCP)CM

Selection of media is based on potential pathogens

Must evaluate specimen type and collection Must evaluate specimen Gram stain to determine if additional media is needed in addition to routine wound culture media

Wound Cultures

Wound Cultures

Routine media to inoculate

Routine media to inoculate

Aerobic media (inoculated with aerobic swab)

Sheep Blood Agar M C k agar MacConkey Chocolate Agar If indicated: PEA or CAN Thioglycollate broth (THIO)

Anaerobic media (inoculated with anaerobic swab)

K Kanamycin, i Vancomycin V i (KV) agar Supplemented blood agar (anaerobic BAP) Ana PEA Thioglycollate broth (THIO)

Common Pathogens in Wound Infections

Common Pathogens in Wound Infections

Eye Bacterial Culture

Pathogens
Staphylococcus aureus Streptococcus pneumoniae Beta hemolytic Streptococcus species Moraxella catarrhalis Moraxela lacunata

Eye Bacterial Culture

Media to Use Pathogens
Haemophilus species Neisseria gonorrheae

Media to Use Chocolate Agar

5% Sheep Blood Agar

Enteric gram-negative rods Pseudomonas species

MacConkey Agar

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Normal Flora in Wound Infections

Common Pathogens in Wound Infections

Ear Bacterial Culture Eye Bacterial Culture

Normal Flora of the Eye includes:

Coagulase negative Staphylococcus species N h Non-hemolytic l ti and d alpha l h h hemolytic l ti streptococci t t i Diptheroids Propionibacterium species

Pathogens
Staphylococcus aureus Streptococcus pneumoniae Beta hemolyitic Streptococcus species Moraxella catarrhalis

Media to Use

5% Sheep Blood Agar

Remember, normal flora will grow on routine culture media for eye cultures

Common Pathogens in Wound Infections

Normal Flora in Wound Infections

Ear Bacterial Culture Ear Bacterial Culture

Pathogens
Haemophilus species

Media to Use
Chocolate Agar

Normal Flora of the Ear includes:

Coagulase negative Staphylococcus species N h Non-hemolytic l ti and d alpha l h h hemolytic l ti streptococci t t i Diptheroids Propionibacterium species

Enteric gram-negative rods Pseudomonas species

MacConkey Agar

Remember, normal flora will grow on routine culture media for ear cultures

Anaerobes

Anaerobic BAP, KV agar, and/or Ana PEA agar

Common Pathogens in Wound Infections

Common Pathogens in Wound Infections

Wound Bacterial Culture

Pathogens
Staphylococcus aureus Group A Streptococcus Group D Streptococcus Enterococcus species

Wound Bacterial Culture

Media to Use Pathogens
Enteric gram-negative rods Pseudomonas species

Media to Use MacConkey Agar

5% Sheep Blood Agar

Anaerobes Anaerobic BAP, KV agar, and/or Ana PEA agar

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Normal Flora in Wound Infections

Wound Cultures

Wound Bacterial Culture

Incubation of Media

Temperature
35 degrees Celsius

No normal flora present but may include skin contaminants Normal Flora of the Skin includes:
Staphylococcus species and Micrococcus species Bacillus species Streptococcus species viridans group Diptheroids Propionibacterium species Yeast

Atmosphere
Ambient air: MAC and THIO CO2 : BAP, CHOC, PEA Anerobically: Supplemented BAP, KV agar, Ana PEA

Time
Aerobically incubated media: 18 24 hours Anaerobically incubated media: 48 hours

Wound Cultures

Wound Cultures

Review

References
Mahon, C.R., Lehman, D.C. & Manuselis, G., Textbook of Diagnostic Microbiology, 3 d Ed 3rd Ed., S Saunders d El Elsevier, i 2007 2007. Wound CultureGeneral Procedure, UNMC CLS Program, CLS 418 & 419

Selection of Culture Media

Based on common pathogens and gram stain

Common Pathogens Normal Flora or Skin Contaminants Incubation of Media

Temperature, atmosphere, time