Pus, Ulcer Material, Skin Specimens: Culture and sensitivity test

Pus, Ulcer Material, Skin Specimens Culture and sensitivity test

Introduction of Pus, Ulcer Material, Skin Specimens

Pus, Ulcer Material, Skin Specimens are common samples for microbiological examination in a tertiary care center set up like gram stain, culture and sensitivity.

Sample collection

Abscess

Container: Aerobic swab moistened with Stuart’s or Amie’s medium

Patient preparation: Wipe area with sterile saline or 70% alcohol. Swab along  leading edge of wound.

Transport to the laboratory: Within 24 hours art room temperature.

Attention

Pus from an abscess is best collected at the time the abscess is incised and
drained, or after it has ruptured naturally. When collecting pus from abscesses, wounds, or other sites, special care should be taken to avoid contaminating the specimen with commensal organisms from the skin. As far as possible, a specimen from a wound should be collected before an antiseptic dressing is applied.

Describe Specimen

For additional investigations
Look for granules: When mycetoma or actinomycosis is
suspected

Culture Pus, Ulcer Material, Skin Specimens

Blood agar
Incubate aerobically
MacConkey agar
Incubate aerobically
Cooked meat medium

For anaerobic bacteria
Subculture at 24 hours , 48 hours, and 72 hours as indicated

Optional
Neomycin blood agar when
anaerobic infection is suspected
Incubate anaerobically up to 48 h
Culture for M. tuberculosis or M. ulcerans
Requires facilities of a
Tuberculosis Reference Laboratory

Examine Microscopically of Pus, Ulcer Material, Skin Specimens

Gram smear
For pus cells and bacteria

Optional steps 
Ziehl-Neelsen smear:When tuberculosis or M. ulcerans disease is suspected
KOH preparation:
When a fungal or actinomycete infection is suspected
Giemsa or Wayson’s smear:
When bubonic plague is suspected
Polychrome methylene blue:
When cutaneous anthrax is
suspected
Dark-field microscopy:
To detect treponemes when
yaws or pinta is suspected

Day 2 and Onwards

Examine and Report Cultures of Pus, Ulcer Material, Skin Specimens
Blood agar and MacConkey agar cultures
Look particularly for:
S. aureus
S. pyogenes
P. aeruginosa
Proteus species
E. coli
Enterococcus species
Klebsiella species
Anaerobes:
C. perfringens
Bacteroides fragilis group
Peptostreptococcus species

Biochemical tests

Depending on the nature of organisms

Antimicrobial susceptibility test

Antimicrobial sensitivity pattern also depends on the nature of organism involvement

On 3rd day

Organism isolated

Antimicrobial sensitivity pattern

Sensitive (S)

Resistant (R)

Intermediate (I)

Possible pathogens of pus

Bacteria
Gram positive
Staphylococcus aureus
Streptococcus pyogenes
Enterococcus species
Anaerobic streptococci
Other streptococci
Clostridium perfringens
and other clostridia
Actinomycetes
Actinomyces israeli
Also Mycobacterium tuberculosis
Gram negative
Pseudonomas aeruginosa
Proteus species
Escherichia coli
Bacteriodes species
Klebsiella species
Pasteurella species
Fungi
Histoplasma
Candida albicans,
mycetoma causing fumgi
Parasites
Entamoeba histolytica
(in pus aspirated from an amoebic liver abscess)

Commensals
Any commensal organisms found in pus are usually those that have contaminated the specimen from skin, clothing, soil, or from the air if an open wound.

Ulcer Material and Skin Specimens 
Possible pathogens
Bacteria
Gram positive
Staphylococcus aureus
Streptococcus pyogenes
Enterococcus species
Anaerobic streptococci
Erysipelothrix rhusiopathiae
Bacillus anthracis
Gram negative
Escherichia coli
Proteus
Pseudomonas aeruginosa
Yersinia pestis
Vincent’s organisms
Mycobacterium leprae

Mycobacterium ulcerans,

Treponema carateum,

 Treponema pertenue.
Viruses
Poxviruses

herpes viruses
Fungi
Dermatophytes (ringworm fungi)
Malassezia furfur
Fungi that cause chromoblastomycosis
Candida albicans

Parasites
Leishmania species
Onchocerca volvulus
Dracunculus medinensis

Commensals
Commensal organisms that may be found on the
skin include:
Gram positive
Staphylococci
Micrococci
Anaerobic cocci
Viridans streptococci
Enterococci
Diphtheroids
Propionibacterium acnes
Gram negative
Escherichia coli
and other coliforms

References

  1. Bailey & Scott’s Diagnostic Microbiology. Editors: Bettey A. Forbes, Daniel F. Sahm & Alice S. Weissfeld, 12th ed 2007, Publisher Elsevier.
  2. Clinical Microbiology Procedure Hand book Vol. I & II, Chief in editor H.D. Isenberg, Albert Einstein College of Medicine, New York, Publisher ASM (American Society for Microbiology), Washington DC.
  3. Colour Atlas and Text book of Diagnostic Microbiology. Editors: Koneman E.W., Allen D.D., Dowell V.R. Jr and Sommers H.M.
  4. Jawetz, Melnick and Adelberg’s Medical Microbiology. Editors: Geo. F. Brook, Janet S. Butel & Stephen A. Morse, 21st ed 1998, Publisher Appleton & Lance, Co Stamford Connecticut.
  5. Mackie and Mc Cartney Practical Medical Microbiology. Editors: J.G. Colle, A.G. Fraser, B.P. Marmion, A. Simmous, 4th ed, Publisher Churchill Living Stone, New York, Melborne, Sans Franscisco 1996.
  6.  Manual of Clinical Microbiology. Editors: P.R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover and R. H. Yolken, 7th ed 2005, Publisher ASM, USA
  7.  Text book of Diagnostic Microbiology. Editors: Connie R. Mahon, Donald G. Lehman & George Manuselis, 3rd edition2007, Publisher Elsevier.
  8. Topley & Wilsons Principle of Bacteriology, Virology and immunology Vol I, II, III, IV & V. Editors: M.T. Parker & L.H. Collier, 8th ed 1990, Publisher Edward Arnold publication, London.
  9. Medical Microbiology-The Practice of Medical Microbiology Vol-2-12th Edn. –Robert Cruickshank
  10. District Laboratory Practice in  Tropical Countries  –  Part-2-   Monica Cheesebrough-   2nd Edn Update
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