Chapter # 5

Urine Examination
5.1. Objective
Examination of urine is indicated in certain renal diseases and diseases
of blood. Sometimes we examine the urine for the presence of certain
parasites, their larvae or eggs in the urine e.g.,
a) Eggs of Schistosoma haematobium, Dioctophyoma renale (kidney worm
of dog), Capillaria plica (bladder worm of dog), Stephanurus dentatus
(kidney worm of pig).
b) Trophozoites of Trichomonas vaginalis.
c) Microfilariae of Wuchereria bancrofti and Onchocerca volvulus.
In areas where schistosoma infection is endemic, the first indirect evidence of
infection is hematuria or proteinuria. Gross hematuria indicates heavy
infection.

5.2. Collection of urine

The urine samples can be collected by one of the following methods.

a) Polythene bags

In large male animals the polythene bags can be applied over urethra and tie
them with the help of rope over the back. When animal urinates the urine is
collected in the polythene bags. It is the method of choice for quantitative
examination of the urine sample.

b) Urethral stimulation

In cows, buffaloes, sheep and goat, one can stimulate the urethra through
vulva, which allows the animal to urinate. Collect the urine sample in clean
and dry container.

c) Per rectum pressure

In large animals, urine can also be collected by applying pressure on the

urinary bladder per rectum.
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d) Through syringe

In small animals, urine can be collected directly from the bladder through
syringe and needle using aseptic conditions.

e) Catheterization

This method is used in small animals.

➢ Clean the genitalia with soap and water and applying antiseptic solution.
➢ Dilate the vagina by using vaginal speculum with left hand and insert the
sterile catheter gently with right hand.
➢ When the catheter is inserted, slight pressure is applied on the abdomen
to press the urinary bladder and urine is collected through catheter.

A fresh or midway urine is preferred for Schistosoma haematobium while

Onchocerca volvulus is usually seen following treatment with
Diethylcarbamizine. Trichomonas vaginalis may be recognized in a fresh
specimen of urine by its jerky motility.

5.3. Preservation of Urine

Normally, the urine sample is examined just after collection. All attempts
should be made to examine the urine sample as soon as after its collection for
proper interpretation. However, when delay is expected, we can use following
methods for preservations:

1) Refrigeration at 4C.
1) Toluene can be added to form a layer over urine.
2) One drop of 40% formaline can be used.
3) Thymol 0.1 gm/10ml urine.
4) One drop of phenol / 10ml of urine.
5) 1ml undiluted formaline (37% formaldehyde) for 100ml urine.
6) 2ml ordinary household bleach for 100ml urine.

5.4. Examination of urine

Two methods are used for urine examination.

a. Sedimentation technique.
b. Syringe filtration technique.

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a. Sedimentation technique
Principle

The parasites being heavier are settled in the sediment by centrifugation and
can be easily isolated.

Materials

➢ Centrifuge machine & centrifuge tubes

➢ Cover slips
➢ Conical flask
➢ Microscopic slides
➢ Pen or marker for labeling
➢ Pipettes

Procedure

➢ Shake the urine well and pour into the conical flask.
➢ Allow the urine to sediment for 1h.
➢ Withdraw the supernatant, transfer the sediment into the centrifuge tube.
➢ Centrifuge at 2000g for 2 minutes.
➢ Place a drop of sediment on a clean glass slide.
➢ Cover it with a cover slip and examine it for the presence of ova.

b. Syringe filtration method

Principle

The eggs of parasites being larger in size than the pores of the filter are
isolated from the sample and are examined after staining.

Materials

➢ Filter holder 13mm or 16mm in diameter

➢ Forceps
➢ Cover slips
➢ Plastic syringe 10ml
➢ Membrane filter12 or 20micron meter, nylon or paper filter.
➢ Microscopic slides
➢ Lugol’s iodine (stock 5% solution)

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Procedure

➢ Place a polycarbonate membrane or nylon filter in a filter holder.

➢ Agitate the urine sample by shaking it gently or by filling and emptying
the syringe twice.
➢ Draw 10ml of the urine into the syringe and attach the filter holder to the
bottom of the syringe.
➢ Keeping the unit level, expel the urine from the syringe into the filter
holder over a bucket or sink.
➢ Carefully unscrew the filter holder.
➢ Draw air into the syringe, reattach the syringe to the holder and expel the
air. This helps to remove excessive urine and also makes sure the eggs if
present are attached to the filter.
➢ Unscrew the filter holder.
➢ Remove the filter with the help of forceps and place it on the microscope
slide with top side up.
➢ Add one drop of lugol’s iodine and wait for 15seconds for the stain to
penetrate the eggs.
➢ Examine the whole filter under the microscope.
➢ Infection loads are recorded as the number of eggs per 10ml of urine. To
estimate the intensity of infection of the sample, divide the no. of eggs
counted by 10. If, less than 10ml was examined then use the following
equation:

No. of eggs / 10ml of the sample = No of eggs counted x 10

X
Where X = no of ml of filtered urine examined

Figure 1. Draw 10ml of urine in a syringe Figure 2. Filter the urine

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Figure 3. Unscrew the filter holder Figure 4. Expel the air through the filter

Figure 5. Stain the filter and examine under microscope

Reuse of filter
➢ Remove the plastic filter immediately after use and soak it overnight in a
1% hypochlorite solution (domestic bleach).
➢ Wash thoroughly with detergent solution and then several tines with
clean water.
➢ Check the filter microscopically to ensure that it is free of parasites
before being reused.

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Precautions

➢ During catheterization, too much pressure should not be applied if any

obstruction is felt.
➢ Do not increase the centrifugation time as this may rupture the ova and
release miracidia.
➢ Process as soon as possible.
➢ Shake the container before pouring in the centrifuge tubes for
sedimentation.
➢ Label the slides or tubes or papers carefully.
➢ Examine the whole specimen as the ova may be very scanty.
➢ Certain crystals, amorphous materials, epithelial or pus cells should not
be confused with parasitic eggs.

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