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Culture of normally sterile fluids


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Culture of normally sterile fluids

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