Unit 5-Urology

A. Objectives
 To be familiar with the way of Collection, Preservation and Dispatch of urine specimen.

 To be familiar with culture procedure of urine sample.

 To perform the sensitivity examination of urine.

 To identify microbes sensitive to particular disease.

B. Content elaboration

5.1 Urine sample collection

While collecting the urine sample, if the animal is urinating the middle stream of the urine
should be collected, if the animal is not urinating we can give pressure on urinary bladder to
make it urine forcefully. We can also use catheter to collect the urine.

5.2 General properties of urine

We cannot store urine at room temperature for a long interval of time because the physical and
chemical properties of urine may alter. If there is the presence of bacteria in urine their number
may rise dramatically. If the bacteria is capable to breakdown the urea PH of urine may rise. If
the urine is alkaline the cast cells may dissolve and may not be visible while analyzing the urine
so fresh urine at morning time should be collected and analyzed.

1.3 Routine examination of urine

Examination of urine can be classified according to
1. Physical examination
2. Chemical examination
3. Microscopic examination
1. Physical examination: on the basis of physical examination, volume of urine, color, smell,
reaction, PH, specific gravity and transparency are examined
 Volume: Volume of urine is influenced by various factors such as; physical
condition of animal, water intake, environmental factors, temperature, feed etc.
volume of urine of different animals are as follows.
Species of animal Volume of urine/day in litre
1. Horse,mule 2-11 (Average 4.7)
2. Cattle/buffalo 8.8-22.6 (Average 14.2)
3. Sheep/goat 0.5-2 (Average 14.2)
4. dog 0.5-2 (Average 14.2)
5. pig 2-6 (Average 14.2)

 Color:

Yellow color of urine is due to presence of urochrome. Higher the urochrome concentration the
color of urine becomes dark yellow and lower the urochrome concentration the color of urine
becomes faint yellow. Most yellow color is suspected due to diabetes, pyometra, jaundice or
kidney malfunctions. Color of urine is also related to specific gravity and volume of urine.

Various color of urine is detected in laboratory. The common color of urine are listed below

 colorless
 yellow
 dark yellow
 pale yellow
 yellowish brown
 greenish yellow
 greenish
 red and milky color
- Dark yellow color is seen in acute nephritis, dehydration and fever.
- Coffee color is seen in case of babesiosis, hypo-phosphotaemia
- Red color is seen due to presence of hemoglobin, phenothiazine,
phenophtalin, azusulamide etc.
- Milky white is due to presence of pus and fat in urine.
- Green color is seen due to degradation of urine sample, phenol poisoning and
intravenous application of methylene blue.
- Orange color of urine is seen in the case of jaundice where urobilinogen is present.
 Smell/Odor:
 Normally urine is light aromatic.
 Pungent odor is received due to the presence of ammonia forming bacteria.
 Putrid sample of urine is received in the case of urinary system infection.
 Fruit smell in urine is received in the case of ketosis.
 PH:
 Urine PH helps to understand the condition of kidneys. PH is analyzed by using
litmus paper, PH meter and reagent strip. If the blue litmus paper turns red the urine
is acidic and if it turns blue the urine sample is alkaline. PH of urine differs according
to
 animal species, feed and water supplement. PH of herbivorous animals is alkaline
whereas carnivorous animal is acidic.

Species PH value
Horse Alkaline 8.0
Cattle/Buffalo Alkaline (7.4-8.4)
Sheep/Goat Alkaline (7.2-8.4)
Pig Alkaline or Acidic depends upon the feed intake
Dog Acidic( 6-7)
Man Acidic (4.6-7) Average 6

 Specific gravity:

Specific gravity is detected to know the filtration capacity of kidney. Specific gravity is analyzed by using
urinometer, refractometer and reagent strip.

Species of animals Specific gravity Average

Horse 1.20-1.050 1.035
Cattle/Buffalo 1.015-1.045 1.035
Sheep/Goat 1.015-1.050 1.030
Dog/Cat 1.020-1.045 1.025
pig 1.005-1.025 1.015

 Transparency:
- Cattle and buffalo: Normally in the case of cattle and buffalo fresh urine is clear
and transparent.
- The urine of horse is thick and cloudy due to the presence of calcium carbonate.
2. Chemical examination of urine:
The chemical examination of urine is done to detect the presence of protein, glucose, ketone,
bile, blood etc in urine.
 Protein test:normally the urine does not contain protein. Protein in urine is seen
during, contraction of muscles, consumption of diet rich in protein, emotional stress,
kidney’s failure and fever
Protein test can be done by:
a. Heat and acetic acid test method
b. Robert’s test
c. Sulfosalicylic acid test
d. Reagent strip test
a. Heat and acetic acid test method:
 Add 2/3 part of urine on 5 ml test tube.
 If the urine sample is alkaline, add some drops of 10% glacial acetic acid and make
the urine acidic.
  Heat the upper part of the test tube.
 Add some drops of 10% acetic acid on urine sample. If the urine gains cloudy
color we can understand that the urine sample contains protein, if the urine shows
no change in color absence of protein can be understood.
 In this method protein and phosphate can be examined.
b. Robert’s test:
 In this method ,concentrated acid helps to coagulate the urinary protein, as a result
the presence or absence of protein in the urine can be detected.
 Place 2 ml of robert’s reagent on a clean test tube.
 Add 2 ml of urine sample (drop by drop) with the help of pipette by touching on the
wall of test tube. (if the urine sample is cloudy, it should be centrifuged before
performing the test)
 A ring is formed on the area of mixing of urine and reagent, if the urine contains
protein (albumin) if the urine does not contain protein the ring will not form.
c. Sulfosalicylic acid test method:
 It is a trusted method to detect the presence of protein in urine.
 If the urine is alkaline add some acid to the urine and make acidic.
 Keep 2 ml of urine sample in a test tube ,add 4-5 drops of 20% sulfosalicylic acid
on the test tube with urine sample.
 Heat the test tube, if the urine becomes turbid the sample is declared to be
protein positive.
d. Reagent strip method:
Protein in urine can also be detected with the help of multistik, combistik etc
 Glucose test:
Normally urine does not contain glucose. In case of cattle/ buffalo, there is less
importance of sugar test. Sugar test has great importance in case of dog. Glucose is seen
in urine in the case of diabetes. To detect the glucose in urine Benedict’s test is quiet
helpful. Benedict’s test:
 Benedict’s reagent is easily available at market
 Keep 5 ml of benedict’s reagent on test tube.
 Add 0.5 ml of urine sample on the test tube and heat the solution for 2 minutes
on Bunsen burner or spirit lamp.
 Be careful that the solution my spill out from the test tube while burning
on Bunsen burner.
 Let the test tube to cool, tally the colour obtained by the solution with the
chart provided on the reagent bottle.
 Ketone test:
Normally the urine of cattle/buffalo does not contains ketone bodies. Ketone body
is composed of acetoacitic acid (20%), acetone(2%) and betahydroxy butyric acid
(78%). Ketosis is the diseases caused by the alteration of fat metabolism due to
 which ketone bodies are present in urine. This disease is mainly seen on lactating
cattle/buffaloes. To detect the ketone bodies rothera’s test is quiet helpful.

Rothera’s test:
 Keep 4 mg of rothera’s reagent on a clean dry test tube.
 Add some urine to dissolve the reagent powder.
 Observe it after 1 minute.
 If the reagent color change into purple color then the presence of
ketone bodies in urine is declared.
 Bile salt
The biles present on urine are urobilinogen, bile salt and bile pigments. Bile pigment
is present on the urine while there is malfunctioning of the liver. Bile salt can be tested by
following way.
Gmelin’s test:
 Take 2 ml of urine sample on clean, dry test tube.
 Add 2 ml of concentrated nitric acid on the urine sample.
 If the samples contain bile salt, it changes into greenish blue color.
 Bile salt is present in urine during obstructive jaundice.
 Blood:
 Keep 2 ml of urine on a test tube.
 Add 2 ml of benzidine saturated solution.
 Mix 1 ml of fresh hydrogen peroxide.
 If the urine contains blood, it changes into blue or green color.

Presence of blood in urine is known as hematuria, presence of hemoglobin in urine is

known as hemoglobin urea.

3. Microscopic Examination of Urine:

Sediment of the urine is used to examine the microscopic examination. In microscope
epithelial cell, pus cell, crystal, erythrocyte etc can be examined. Spermatozoa , parasites,
fungus and tumor cells can also be examined by this method.
Reference:

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