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Culture of normally sterile fluids

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Microbiology is essentially hidden knowledge to rule out disease,however together we can

Microbiology is essentially hidden knowledge to rule out disease,however together we can

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  • 1. CULTURE OF NORMALLY STERILE FLUIDS
    • Lusubilo Malakbungu(BMLS)
    Lusu2012
  • 2. Normally sterile Fluids
    • These are fluids that do not contain commensals. Any marked presence of commensals can lead to progression of disease in these fluid and site in particular.
    Lusu2012
  • 3. Some diseases occuring in normal sterile sites
    • Amnionitis -inflamation of the amnion(fetal sac).Due to contamination of vaginal flora or instrumentation during antenatal medical procedures.
    • Pericaditis -inflamation of the pericardium(membrane enveloping the heart).
    • Peritonitis -inflamation of the peritoneum.
    • Empyema -collection of pus in pleural cavity
    Lusu2012
  • 4. .
    • Septic athritis- pyogenic infection of a joint. Spread by haematogeneous spread.
    • Bursitis- inflamation of bursa(small fluid filled sac of fibrous tissues lined with synovial membrane)
    • Meningitis- inflamation of the meninges in the brain. Agents can be Haemophilus influenza, Streptococcus pneumoniae.Neisseria meningitidis.
    Lusu2012
  • 5. .
    • CULTURING PROCEDURES FOR SOME NORMAL STERILE FLUIDS
    Lusu2012
  • 6. 1.Cerebral Spinal Fluid
    • Collection should be done by experienced personnel.
    • 5-10ml of CSF should be collected in two sterile tubes.
    • Disinfection of the skin is mandatory.
    • Cells of CSF disintergrate rapidly,so they should be processed immediately without delay.
    • Lumbar puncture done between L3 and L4
    Lusu2012
  • 7. Lusu2012
  • 8. .
    • If transport to laboratory is not possible,inoculate CSF aseptically into Trans-Isolate (T-I)Medium and held overnight at 35C
    • MACROSCOPIC
    • Cloudy appearance give a clue of pyogenic infections.
    • Gram staining
    • CSF should be centrifuged and the sediments smeared for staining.
    Lusu2012
  • 9. Lusu2012
  • 10. CULTURE
    • The drop of sediments after centrifugation is used for culture.
    • MEDIA:Blood agar,Chocolate Agar(Tryptone Soy Agar) .Suspected organisms for this culture include N.meningitidis,H.influenza,S.pneumoniae.
    • -SDA-used for any clue of yeast cells in gram stain.
    Lusu2012
  • 11. .
    • MCA-when neonatal meningitis is suspected,for isolating Escherichia coli also Listeria monocytogenes
    • RESULTS
    • C olonies could be of:Neisseria meningitidis(in Blood Agar & chocolate agar)
    • S.pneumoniae,H.influenza(CA),Cryptococcus neoformans(SDA).
    Lusu2012
  • 12. Lusu2012
  • 13. Confirmation of colonies
    • H.Influenzae will grow on CA and satellite colonies in the vicinity of staphylococcus streak.
    • H.Influenza type b-antiserum slide agglutination.
    • On CA rapid oxidase positive is suggested to be meningococci.
    • N.Meningitidis antisera(A,B,C) in slide agglutination
    Lusu2012
  • 14. Lusu2012
  • 15. OTHER STERILE SITES
    • Pericardial effusion
    • This infections pericaditis can be:-
    • Purulent caused by bacteria.
    • Benign can be due to viruses
    • Granulomatous which are caused by mycobacterium tuberculosis.
    • Wide range of bacteria isolated for purulent include Streptococcus spp,Neisseria spp,Haemophilus.
    Lusu2012
  • 16. BONE MARROW
    • Should be inoculated into commercially available Nutrient broth(e.g Brain-Heart infusion broth or TSB)
    • Common isolate is S.aureus.
    Lusu2012
  • 17. OTHER EFFUSIONS
    • Synovial
    • Pleural
    • Ascitic (peritoneal)
    Lusu2012
  • 18. Collection
    • Is done by a well trained medical officer.
    • After aspiration ,aseptically dispense the fluid as follows;
    • 2-3ml into a dry,sterile,screw-cap tube or bottle to observe for clotting.
    • 9 ml into a screw-cap tube or bottle which contains 1ml of sterile tri-sodium citrate, mix well for anticoagulation.
    Lusu2012
  • 19. Microscopic Examination
    • Gram smear-look for pus cells and bacteria.
    • Ziehl-Neelsen
    • Look for AFB
    • Wet preparation for crystals:
    Lusu2012
  • 20. CULTURE
    • Blood agar (incubate aerobically)
    • Chocolate agar(incubate in C02
    • MacConkey agar
    • Incubate aerobically.
    • For suspected Tuberculosis-required facilities of tuberculosis should be used.
    Lusu2012
  • 21. Report cultures
    • Look particularly for S.aureus,s.pyogenes,s.pneumoniae,H.influenzae,Neisseria spp,Enterobacteria.
    Lusu2012
  • 22. Lusu2012
  • 23. Lusu2012
  • 24. references
    • Isolation of Agents from Normally Sterile Sites ;CDC covers 2003.
    • Investigations of Fluid from Normally sterile sites; Issued by Standards Unit, Department for Evaluations, Standards and Training, Centre for Infections.
    • Cheesebrough M.District Laboratory Practice in Tropical Countries, Part 2 Second Edition
    Lusu2012