Collection and Processing of Body Fluids

Collection and transport of effusions

Series of synovial, pleural, pericardial, peritoneal, or hydrocele fluid is achieved by way of a scientific officer.

In a hospital with a microbiology laboratory

After aspiration, aseptically dispense the fluid as follows:

• Take 2–3 ml into a dry, sterile, screw-cap tube or bottle to look at for clotting.
• Take 9 ml right into a screw-cap tube or bottle which carries 1 ml of sterile tri-sodium citrate blend the fluid with the anticoagulant.

Remember:Tri-sodium citrate prevents clotting, specifically of exudates. The sterile citrated pattern may be used to estimate cell numbers, protein awareness, and for microscopy and subculture.

Label, and as soon as viable supply the samples with a finished request form to the laboratory.

In a health post for dispatch to the laboratory

After aspiration, aseptically dispense the fluid as follows:

• Take five ml into a bottle of sterile thioglycollate broth and blend.
• Take nine ml right into a screw-cap tube or bottle which includes 1 ml of sterile tri-sodium citrate.
• Mix the fluid with the anticoagulant,If any fluid stays, dispense into a dry, sterile, screw-cap tube or bottle, and take a look at for clotting.
• Then,label every box with the date and the affected person’s name, range, and fitness center.
• And transport the samples with a finished request form to attain the microbiology laboratory inside some hours. The inoculated thioglycollate broth must be kept in a heated environment, but now not over 37 _C or in direct daylight.

Lab exam of effusions

First day

Describe the type of the specimen

Document:

• Shade of the effusion
• Whether it is clean, cloudy, or purulent (like pus)
• Whether or not it contains blood
• Whether or not it's miles clotted (sample without anti-coagulant)

Purulent effusion

While the specimen is pus or markedly cloudy, take a look at and document a Gram-stained smear as soon as viable. Proceed to examine the specimen as for blood-stained effusion.culture the specimen and observe a Gram-stained smear word.When the specimen isn't pus or a bloodstained effusion, transfer about 1 ml of the well-mixed citrated sample to a separate tube or bottle .Use this to estimate cell numbers and protein attention . This will avoid contaminating the rest of the sample which can be required for culture. Whilst the non-citrated sample does now not include clots, this must be used for the cellular rely upon and protein in preference to the citrated sample.

Have a look at the fluid for cells

Estimate the number of white cells inside the fluid the usage of the technique described for c.s.f. Report whether the cells are mainly polymorphonuclear neutrophils (pus cells) or lymphocytes. Tuberculous effusions comprise specially lymphocytes and frequently plasma cells (it's miles rare to find AFB in the fluid).

Be aware: A transudate may additionally incorporate a few cells, while an exudate typically incorporates many cells.

Estimate the protein

Dilute the fluid 1 in one hundred in physiological saline (0.1 ml effusion mixed with nine.9 ml saline). Estimate the general protein using the approach defined for measuring general protein in c.s.f. Multiply the end result with the aid of a hundred.A transudate typically consists of much less than 30 g/l (3 g/dl) of protein whereas an exudate includes more than 30 g/l.

Observe: An exudative pleural effusion containing lymphocytes without organisms visible within the Gram smear is located with tuberculosis. The subsequent desk summarizes the results of a few exams which may be finished in district laboratories to differentiate transudates from exudates.

Culture the specimen

Subculture the fluid when it incorporates a variety of white cells and extra than 30 g/l of protein, or when it seems blood-stained . Centrifuge the citrated pattern in a sterile tube at excessive pace for approximately 20 mins to sediment the microorganism. Do away with the supernatant fluid (do now not discard) and resuspend the sediment. subculture the sediment as follows:

On Chocolate agar, blood agar, and MacConkey agar

• Inoculate the sediment on chocolate (heated blood) agar, blood agar, and MacConkey agar.
• Incubate the chocolate agar plate in a carbon dioxide enriched ecosystem at 35–37 _C for up to forty-eight hours, checking for the increase after overnight incubation. Incubate the blood agar plate and MacConkey agar plate aerobically at 35–37 _C for up to 72 hours, examining for the increase after in a single day incubation.

After two days or extra

Take a look at and document the cultures of Chocolate agar, blood agar, and MacConkey agar cultures

appearance mainly for colonies that could be:

• Staphylococcus aureus,
• Streptococcus pyogenes
• Streptococcus pneumoniae
• Haemophilus influenzae
• Enterobacteria
• Pseudomonas aeruginosa
• Neisseria species

Something more

Moist instruction to hit upon crystals when gout or pseudogout is suspected (generally in males) .Whilst the fluid is from a joint, transfer a drop of the sediment from the centrifuged fluid (citrated pattern) to a slide, and cowl with a cowl glass. study the guidance using the 10_ and 40_ targets with the condenser iris closed sufficiently to offer maximum contrast. search for colorless, extracellular and intracellular (interior white cells) crystals:

• Monosodium urate crystals are needle-like in shape and measure 8–10 _m in length. They may be found in effusions from sufferers with gout.
• Calcium pyrophosphate crystals measure up to twenty-five _m in duration, are rod-fashioned, and might have a line running through them. They can be found in effusions from sufferers with pseudogout.

Gout: High serum urate ranges are commonly found in sufferers with gout. everyday levels are observed in sufferers with pseudogout.

Cytology smear whilst malignancy is suspected

Make two thin smears of effusion sediment and at the same time as nonetheless moist, restoration the smears in a box of 95% v/v ethanol for 20 minutes. send the smears to a Cytology Laboratory for unique staining and exam for malignant cells.

Possible pathogens

Synovial fluid

(Synovitis and Infective arthritis)

Gram-positive

Staphylococcus aureus

Streptococcus pyogenes

Streptococcus pneumoniae

Anaerobic streptococci

also Mycobacterium tuberculosis.

Gram negative

Neisseria gonorrhoeae

Neisseria meningitidis

Haemophilus influenzae

Brucella species

Pleural and pericardial fluids

(Empyema and Purulent Pericarditis)

Gram-positive

Staphylococcus aureus

Streptococcus pneumonia

Streptococcus pyogenes

Actinomycetes Klebsiella traces

Gram negative

Haemophilus influenzae

Bacteroides

Pseudomonas aeruginosa

also, Mycobacterium tuberculosis, fungi, and viruses particularly coxsackie B virus.

Ascitic fluid

(Ascites and Peritonitis)

Gram-positive

Enterococcus species

Streptococcus pneumonia

Staphylococcus aureus

Streptococcus pyogenes

Streptococcus agalactiae

Gram negative

Escherichia coli

Klebsiella Species

Bacteroides

Pseudomonas aeruginosa

Different enterobacteria

also Mycobacterium tuberculosis and Candida species.

Hydrocele fluid

On occasion Wuchereria bancrofti ,microfilariae and not often Brugia species can be observed in hydrocele fluid.

References:

D greenwood, Slack RCB and J Peutherer.Medical microbiology.2001.

JG College, AG Fraser and BP Marmion.Practical Medical microbiology.Fourteenth Edition. Churchill Livingstone, 1996.

JP Micheal, ECS Chan and NR Krieg.Microbiology.Fifth Edition. Delhi: Mcgraw-hill, 1993.

M Cheesbrugh.Medical laboratory manual for tropical countries.London, 2007.