Culture of normally sterile fluids

24
CULTURE OF NORMALLY STERIL FLUIDS Lusubilo Malakbungu(BMLS) Lusu2012

description

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

Transcript of Culture of normally sterile fluids

Page 1: Culture of normally sterile fluids

CULTURE OF NORMALLY STERILE FLUIDS

Lusubilo Malakbungu(BMLS)Lusu2012

Page 2: Culture of normally sterile fluids

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

Page 3: Culture of normally sterile fluids

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

Page 4: Culture of normally sterile fluids

.

• 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

Page 5: Culture of normally sterile fluids

.

CULTURING PROCEDURES FOR

SOME NORMAL STERILE FLUIDS

Lusu2012

Page 6: Culture of normally sterile fluids

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

Page 7: Culture of normally sterile fluids

Lusu2012

Page 8: Culture of normally sterile fluids

. 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

Page 9: Culture of normally sterile fluids

Lusu2012

Page 10: Culture of normally sterile fluids

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

Page 11: Culture of normally sterile fluids

. MCA-when neonatal meningitis is

suspected,for isolating Escherichia coli also Listeria monocytogenes

RESULTS

Colonies could be of:Neisseria meningitidis(in Blood Agar & chocolate agar)

S.pneumoniae,H.influenza(CA),Cryptococcus neoformans(SDA).

Lusu2012

Page 12: Culture of normally sterile fluids

Lusu2012

Page 13: Culture of normally sterile fluids

Confirmation of coloniesH.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

Page 14: Culture of normally sterile fluids

Lusu2012

Page 15: Culture of normally sterile fluids

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

Page 16: Culture of normally sterile fluids

BONE MARROW

Should be inoculated into commercially available Nutrient broth(e.g Brain-Heart infusion broth or TSB)

Common isolate is S.aureus.

Lusu2012

Page 17: Culture of normally sterile fluids

OTHER EFFUSIONS

Synovial Pleural Ascitic (peritoneal)

Lusu2012

Page 18: Culture of normally sterile fluids

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

Page 19: Culture of normally sterile fluids

Microscopic Examination

Gram smear-look for pus cells and bacteria.

Ziehl-Neelsen

Look for AFB

Wet preparation for crystals:

Lusu2012

Page 20: Culture of normally sterile fluids

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

Page 21: Culture of normally sterile fluids

Report culturesLook particularly for

S.aureus,s.pyogenes,s.pneumoniae,H.influenzae,Neisseria spp,Enterobacteria.

Lusu2012

Page 22: Culture of normally sterile fluids

Lusu2012

Page 23: Culture of normally sterile fluids

Lusu2012

Page 24: Culture of normally sterile fluids

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