Urine Examination

&
Analysis

Assigned by: Dr.Javeria Khan

Presented by: Dr.Noor-ul-Ain Sarwar
 Contents
1. Collection of sample and preservation.
2. Gross Examination.
3. Determination of specific gravity.
4. Biochemical Analysis.
5. Microscopic Examination.
6. Disease Interpretations
 Introduction
• Urinalysis a very useful tool to evaluate healthy and
diseased animals. It provides valuable information about
the urinary system. There are so many examples of
diseases in which specific urine picture can be seen. for
example:
A. Kidney diseases:
 Abnormal specific gravity Proteinurea.
 Cast
 Leukocyte
 Erythrocytes
B. Bladder infection:
 Proteinuria
 Leukocytes
 Bacteria
C. Neoplasia:
 Exfoliated neoplastic cells
 Hematuria
D. Liver diseases:
 Billirubinuria
 Altered urobilinogen
 Bilirubin crystals
E. Hemolysis:
 Post parturient hemoglobinuria
 Bovine bacillary hemoglobinuria
 Anthrax
 Increased urobilinogen
F. Diabetes mellitus:
 Glycosuria
 Increased volume, increased specific gravity
 Ketonurea
G. Diabetes insipidus:
 Decreased specific gravity
H. Acidosis
I. Alkalosis
Collection of Urine
 1. Collection of urine sample
Collection can be done either by clean catch
method,catheterization or through cystocentesis.
Sample size: 15- 20mL
Best time for analysis:
A fresh urine sample is preferred for
analysis.Ideally urinalysis should be performed
within 30 min of sample collection.If delay of
examination then following changes takes place:
Urea is converted into ammonia that makes the
sample alkaline.
Formed elements(cells,casts)are dissolved.
Precautions while collection of urine:
 Morning samples are most likely to contain
constituents of diagnostic significance.
 Fluid consumption during the day dilutes the urine
resulting in decreased specific gravity.
 Collect mid stream urine.
 In case of diabetes mellitus, sample should be
collected 2 hours after feeding and fasting.
 For nephritis,use only morning samples.
 Direct collection is the preferable method in large
animals.although catheterization and cystocentesis
provide high quality of uncontaminated sample
but are associated with tissue trauma of varying
degree.
Preservation:
Store samples in refrigerator at 8 degree C.(warm
at room sample before analysis).
Precautions regarding refrigeration:
Maximum upto 12 hours
It slightly increases specific gravity.
It also interferes with tests using enzymes for
reaction.
 Chemical preservation
Certain chemicals are used with limitations.
a. Toluene:
Quantity: 2ml/100ml of urine for 24 hours.
Only cover urine surface, don't dissolve in urine.
Limitation :Interferes with ketone bodies
determination.
b. Thymol:
Quantity: a small lump can preserve for several
days.
Limitation: it gives false positive protein reaction.
c. Formaline:
Quantity: 1drop of 40% formalin for 30ml urine for
24 hours.
Limitation: It interferes with glucose reaction.
c. Metaphosphoric acid:
When ascorbic acid is to be determined from the
urine. It is added with a ratio of 1:5 ( 1 part 10%
aqueous solution of metaphosphoric acid and 5
parts of urine sample.
Collection
 2. Gross Examination

a) Volume:
Normal values
species in liters
Horse 4.7
Cattle 14.2
Sheep, Goat 0.9
and Dog
 Interpretation
i. Increased volume-Polyurea:
Physiological:
 Increased water consumption
 Diuretics
 Parenteral fluid therapy
Pathological:
 Chronic progressive renal failure
 Diabetes mellitus
 Diabetes insipidus
 Chronic pylonephritis
  Pyometra
ii. Decreased volume-oligouria:
Physiological:
 Less water intake.
 High environmental temperature.
 Panting
 Dehydration
Pathological:
 Acute renal disease
 Urolithiases
 fever
 Shock
 Severe nephritis
 Edema
 b. Color:

Colour of the urine is due to the concentration

of urochromes.Always consider color in
association of volume and specific gravity of
urine.
Normal Colour:
Freshly voided urine is clear and may range in
color from light pale yellow to amber(gold)or
straw Colour except horses which have turbid
color urine due the presence of calcium
carbonate crystals and mucin.
 Interpretation
Various color could be:
i. Less to pale yellow:
 End stage renal disease
 Increased uptake of water
 Diabetes insipidus
 Hyperadrenocorticism
ii. Dark yellow to yellow brown:
 Acute nephritis
 Dehydration
 Bilirubin
iii. Yellowish brown;
 In birds ,yellowish green urates indicates hemolysis
or liver disease.
 Bilirubin
iv. Red:
 Hematuria
 Hemoglobinuria
v. Brown to brownish black:
 Hemoglobin(hemoglobinuria,post parturient
hemoglobinuria,bacillary hemoglobinuria)
 Myoglobin(Monday morning disease)
 Melanin
vi. Green:
 Biliverdin
 Phenol poisoning
 c. Odor Interpretation
 Ammonia-like : Urea-splitting bacteria
 Foul, offensive : Old specimen, pus or
inflammation
 Sweet : Glucose
 Fruity : Ketones
 Maple syrup-like: Maple Syrup Urine Disease
 d. Color:
 Colorless Diluted urine
 Deep Yellow Conc. Urine, Riboflavin.
 Yellow-Green Bilirubin / Biliverdin
 Red Blood / Hemoglobin
 Brownish-red Acidified Blood (Actute GN)
 Brownish-black Homogentisic acid (Melanin)
 3. Specific Gravity

The ability of kidneys to concentrate the urine.

Determination:
It can be determined by the use of refractometer
or urinometer.the steps are as under:
 Temperature of the urine must be 20-25°C.
 cylinder used for floatation of urinometer
should be large enough in diameter so that
urinometer can flow in it.
.
 Place the urinometer in cylinder containing
the urine .Rotate it to prevent its touching to
the sides.
 Read the scale on the bottom of urinometer
and record it in decimals.
Urinometer
Refractometer
Refractometer
Species Specific Gravity
Horse 1.020 – 1.050

Cattle 1.025 – 1.045

Sheep & Goat 1.015 – 1.024

Dog 1.015 – 1.045

Birds 1.005 – 1.020

 Interpretation
i. Increased specific gravity:
 Acute interstitial nephritis
 Cystitis
 Liver failure
 Diabetes mellitus
 Glomerulonephritis
ii. Decreased specific gravity:
 Chronic interstitial nephritis
 Diabetes insipidus
 Pylonephritis
 uremia
Chemical Analysis
 3. Biochemical Analysis
For biochemical analysis urine must be
uncentrifuged.
a. pH:
i. Acidic pH Interpretation:
 Normal in carnivores.
 Nursing calves & foals.
 Excessive diet in protein.
 Hypokalemia.
ii. Alkaline pH Interpretation:

 Normal in herbivores
 Stale urine sample becomes alkaline.
 Cystitis.
 Normal pH
Species pH

Horses 8

Cattle 7.4-8.4

Sheep & Goat 7-8.2

Dog & Cat 5.5-7

Birds 6-8
 b. Protein Determination:
For Protein determination:
a) Reagent strips(dip sticks)
b) Acid prepitation Tests:
i. Nitric Acid Precipitation Test
OR Robert’s Tests:
Principle:
Precipitation of protein occur by concentrated acid
Procedure:

 Take 2ml of Robert’s reagent in a test tube.

 Place 2ml of urine.
 Wait for few minutes.
Result:
A positive test is indicated by a white ring
at the zone of contact of 2 fluids.
Interpretation of Protein determination
 Hemoglobinuria.
 Myoglobinnuria.
 Pylonephritis.
 Cystitis Urolithiases
 Inflammation,
 Hemorrhage,
 Glomerular disease.
 c. Glycosuria determination
Now a days strips and glucometers are available.
Chemical method is Benedict’s test.
Benedict’s test:
Principle:
It depends upon the reducing sugars present in
the urine to react with copper sulphate to
reduce cupric ions to cuprous oxide giving
color.
Composition of Benedict’s Reagent
Copper sulphate 17.3 g

Sodium citrate 173 g

Sodium carbonate 100 g

Distilled water 1000 mL

(To make volume)
Procedure:
 Take 5mL of Benedict’s reagent in a test tube.
 Add 8 drops of urine to the reagent.
 Mix the 2 fluids.
 Heat it with constant shaking till boiling.
• Result:

Positive Blue color

Negative Orange to
brick red or
brown
 Interpretation
 Hyperglycemia
 After general anesthesia
 Chronic liver diseases
 Enterotoxaemia in sheep.
 d. Ketonurea Determination
Ross test:
Principle:
It is based on that the sodium nitroprusside is
decomposed to:
 Sodium ferrocyanide
 Sodium nitrate
 Ferric hydroxide
Results:
Purple coloration
Procedure:

 Place half inch layer of powdered reagent in

test tube.
 Add 5mL of urine.
 Agitate the 2 components in the test tube.’
 Overlay 1-2 ml of ammonium hydroxide over
the mixture.
 Wait for 4-5 minutes.
 Development of purple color indicates the
presence of ketone bodies in urine.
Interpretations

 Diabetes mellitus
 High fat diet
 Starvation
 Impaired liver functions
 After ether chloroform anesthesia
 Milk fever
 e. Hematuria detection
Benzidine test:
 Take 2mL of glacial acetic acid in a test tube.
 Add small amount of Benzidine reagent.
 Add 1 ml of urine
 Add 1 mL of fresh hydrogen per oxide.
 Wait for 5 minutes.
Result:
• Green or blue color development.
Interpretation
• Acute nephritis
• Urolithiases
• Cystitis
• Tumor of the urethra
• Severe infections like, anthrax,
leptospirosis,infectious canine hepatitis.
• Chemicals like copper, mercury or phenol
poisoning.
• Parasites like Dicroflaria immitus,Dictophyma
renale,Capillaria plica.
 f. Billirubinuria determination
Foam test:
Procedure:
 Take 1-2 mL of urine in a test tube.
 Shake it vigorously.
Result:
• Appearance of yellow, greenish yellow or
brown colour foam above the surface of urine
indicates presence of Bilirubin.
Interpretation:

 Infectious canine hepatitis,leptospirosis.

 Neoplasia
 Obstruction of bile duct.
 Jaundice
 Calcium Determination
Sulkowitch test:
Calcium present in urine reacts with sulkowitch
reagent ,ppt in the form of calcium oxalate.
Procedure:
 Take 5mL distilled water & add 5mL urine in
1 test tube as control.
 In another test tube,mix equal amount of urine
& sulkowitch reagent.
Result:
 Compare the 2 test tubes in light after 2-10 min.

Interpretation:
Increased:
 After Ca administration.
 Hyperthyroidism
 Hypervitaminosis
Decreased:
 In bovines, it is not reliable.
 In canines,pre-renal tetany.
 hypothyroidism
 5) Microscopic examination
Purpose:
 Recognition of cells for urinary tract
infections.
 Exfoliative cytology of tumors.
Procedure;
 Centrifuge sample @ 1500 rpm for 2-3 min.
 Pour off the supernatant.
 Place a drop of sediment on slide and cover it with
a cover slip.
 Observe the slide @ 10x and 40x.
Result variations:
i. Voided sample: more cellular, bacterial
contamination.
ii. Catheterized sample: increased transitional cell
content, iatrogenic hemorrhage.
iii. Cystocentesis: least extraneous contamination,
more specific for changes in the tract,
Interpretations:
• Epithelial cells in neoplasia diagnosis.
• more than 5RBCs/HPF indicate Hematuria.
• Leukocytes indicate infection(pyouria).
• More than 5/HPF.
• Elongated structures like casts indicate presence
of Urolithiases.
• 10,000 bacterial rods/ml and >100,000 bacterial
cocci/mL of urine are required to consistently
find bacteria in a urine sample using light
microscopy. and readings are normally below
this.
 Some of the drugs that excreted in urine also
appear in crystals.e.g:
 Sulfonamide crystals spherical with spikes.
 Ampicillin crystals form long needle lik
arrays.
 Calcium oxalate crystals are like colorless
squares indicate:
 Urolithiases
Ethylene glycol toxicosis
Cytological Examination
• Staining:

– Papanicolau
– Wright’s
– Immunoperoxidase
– Immunofluorescence
 Staining:
WRIGHT STAIN PROCEDURE:
 Make a air dried smear.
 Fix it in methanol for 30 sec.
 Take a disposable pipette and flood the Wright Stain
on the appropriately labeled slides.
 Wait for 3 min.
 Place 1ml oxidizing Wright Stain and Wright Stain
Buffer Mixture on Wright stained slides laying on
slide rack (Displacing the Wright Stain off the slides
with the pipette filled with Wright Stain/buffer
mixture and viewing a metallic sheen on the top of
slides.)____
 Staining:
 Wait for 6 min.
 Place slides in Wright Stain Buffer for 1.5
minutes.
 wait for 1.5 minutes.
 Rinse, dry and examine under oil immersion
lens,100x.
Parasites
Capillaria plica
Dioctophyme renale.
Trichuris
Casts
RBCs Cast - Histology
RBCs Cast
WBCs Cast
Tubular Epith. Cast
Tubular Epith. Cast
Granular Cast
Hyaline Cast
Waxy Cast
Fatty Cast
Crystals
Calcium Oxalate Crystals
Calcium Oxalate Crystals
Triple Phosphate Crystals
Urate Crystals
Leucine Crystals
Cystine Crystals
Bilirubin
Ammonium Biurate Crystals
Cholesterol Crystals
 Cytology

carcinoma
Cytology: Polyoma (Decoy Cell)
Cytology: Squamous Cell Ca.
Cytology: Renal Cell Ca.
Cytology: Prostatic Carcinoma
Cytology
WBCs
Cytology: Normal
Cytology: Normal
Cytology: Reactive
Cytology: Reactive
Tubular Epithelial Cells
WBCs
RBCs
Cocci
Hematuria
Transitional Cells
Oval Fat Body
Transitional Cells
LE Cell
Squamous cell
Cytomegalovirus
Yeasts
Yeasts
Bacteria
Amorphous Substance
Bacilli
Mucous
 Interpretations of Urine Analysis
 Proteinuria
 Casts & cells
 Hematuria
 Hemoglobinuria
 Myoglobinuria
 Pyuria
 Bacteriuria
 Crystalluria
 Glycosuria
 Ketonuria
 Parasites
6.Interpretation Of
Diseases of
Urinary System
Common Findings in:
Acute Tubular Necrosis
Microscopic: Glucose

Renal tubular Bilirubin

Ketones
 epithelial cells
Decreased S.G.
Pathological casts. +/- Blood

pH

+/- Protein

Urobilinogen
Common Findings in:
Acute Glomerulonephritis
Microscopic: Glucose
Glucose

Erythrocytes (dysmorphic) Bilirubin

Bilirubin
Ketones
Erythrocyte casts Specific Gravity
Gravity
Mixed cellular casts Blood
Blood Increased
pH
Protein Increased
Urobilinogen
Urobilinogen
Nitrite
Leukocyte
Leukocyte Esterase
Esterase
Common Findings in:
Chronic Glomerulonephritis
Microscopic: Glucose
Glucose

Pathological casts Bilirubin

Ketones
Ketones
(broad waxy casts, RBCs)
Specific
Specific Gravity
GravityDecreased
Blood Increased

pH
pH
Protein
Protein Increased

Urobilinogen
Nitrite
Nitrite
Leukocyte Esterase
Esterase
Common Findings in:
Acute Pyelonephritis
Microscopic: Glucose
Glucose

Bacteria Bilirubin
Ketones
Ketones
Leukocytes Specific
Specific Gravity
Gravity
Leukocyte, granular, and Blood

waxy casts pH
pH
Protein
Protein Trace
Renal tubular epithelial
Urobilinogen
cell casts Nitrite
Nitrite Positive
Leukocyte Esterase
Esterase
Common Findings in:
Nephrotic Syndrome
Microscopic: Glucose
Glucose

Oval fat bodies Bilirubin

Ketones
Ketones
Fatty casts Specific
Specific Gravity
Gravity
Waxy casts Blood
pH
pH
Protein
Protein ++++

Urobilinogen
Nitrite
Nitrite
Leukocyte Esterase
Esterase
Common Findings in:
Eosinophilic Cystitis
Microscopic: Glucose
Glucose

 Numerous eosinophils Bilirubin

Ketones
Ketones
(Hansel’s stain)
Specific
Specific Gravity
Gravity
 NO significant casts. Blood +

pH
pH
Protein
Protein
Urobilinogen
Nitrite
Nitrite
Leukocyte Esterase
Esterase
Common Findings in:
Urothelial Carcinoma
Microscopic: Glucose
Glucose
 Malignant cells on Bilirubin

urine cytology Ketones

Ketones
Specific
Specific Gravity
Gravity
(urine sample should Blood +
be submitted pH
pH
separately to Protein
Protein

cytology, Urobilinogen
Nitrite
Nitrite
void or 24 hrs.) Leukocyte Esterase
Esterase
 Bacterial Cystitis:

 Urinalysis often shows increased protein and

hemoglobin
 Increased numbers of WBC, RBC, and/or
bacteria are consistent with cystitis.
Urine Culture
 Urine Culture
Purpose:
 To identify the specific infectious agent.
 Antibiotic sensitivity test.
 Urine Culture
 Media Descriptions:
 C.L.E.D. (Cystine Lactose Electrolyte
Deficient Agar) is a non-selective medium that
supports the growth of Gram (+) and Gram (-)
species, specifically for enumeration of
bacteria in urine.
URINALYSIS REPORT: