This document lists various infectious and non-infectious diseases along with information about sample collection and diagnosis for many of them. It provides details on diseases like leptospirosis, rabies, babesiosis, theileriosis, trypanosomiasis, demodicosis, scabies, canine distemper, pyometra, fascioliasis, milk fever, ketosis, hypomagnesemia tetany, and more. For each disease, it specifies the appropriate sample to collect (e.g. blood, skin scraping) and describes clinical pathology tests and methods used to diagnose the condition.

1. ICAR- INDIAN VETERINARY RESEARCH INSTITUTE
IZATNAGAR- 243122

2.  Leptospirosis
 Rabies
 Babesiosis
 Theileriosis
 Trypanosomiasis
 Demodicosis
 Scabies
 Canine Distemper
 Pyometra In Bitch
 Fasicioliasis
 Milk Fever In Bovine
 Ketosis In Bovine
 Hypomagnesium Tetany
 Acidic Indigestion In Ruminents
 Alkaline Indigestion In Ruminents
 Cyanide Poisoning In Ruminents
 Nitrate/Nitrite Poisoning In
Ruminents
 Diabetes Mellitus
 Nephritis
 Traumatic Reticulo-peritonitis (TRP)
Infectious And Non Infectious Diseases

3. Leptospirosis In Dogs
 SAMPLE Collected
 whole blood/serum for serogical
tests(blood must be collected during
early febrile stage)
 Mid stream urine for dark field
examination.
 CSF
 Preferred collection container:
 3 mL lavender-top (K2 EDTA) tube
 Sample required:
 1 mL EDTA whole blood
(preferred)
 or 5 mL urine

4. Commont Test and Conditions
 CBC : lymphocytosis, anemia, neutrophilia, and
thrombocytopenia
 Liver function test: Elevated liver enzymes (e.g., alanine
aminotransferase, aspartate aminotransferase, alkaline
phosphatase) and bilirubin , SGPT, SGOT.
 Kidney function test : elevated BUN and creatine
 Other findings can include electrolyte abnormalities (e.g.,
hyponatremia, hypokalemia, hypochloremia, and/or
hyperphosphatemia).
 Urinalysis : shows protenuria, pyruia, and often microscopic
hematuria.
 Hyaline and granular casts may also be present dring the first
week of illness.

5. Specific Test For Diagnosis Leptospirosis
 Dark Field Microscopy (DFM) : Visualize Leptospirosis
 Microscopic Agglutination Test (MAT) : Gold Standard
 Polymerase Chain Reaction (PCR) : Successful in detecting
Leptospira DNA in serum and urine samples of patients

8. RABIES
Etiology : family – Rhabdoviridae, genus- Lyssa virus
Sample collected :
 saliva and ocular discharge in sterile vial on ice
 Corneal impression smears.

9. Corneal test or saliva test
• This is based on the demonstration of rabies virus antigen in the
corneal epithelium and the saliva of the patient.
• This is the only test that can be done prior to death.
Procedure :
 (A) By Rai Choudhary And Thomas 1971
1. Hold a clean greese free microscopic slide by the one hand.
2. Retract both the eyelids by the thumb and index finger of the
other hand.
3. Pass one end of the microscopic slide on the cornea firmly and
remove it.
4. Repeat this process for several times till a definite smear is
obtained.
5. Prepare at least four corneal smears from each eye.
6. Make the smears air dry.

10. (B). Identification of rabies antigen in smears of saliva by
fluorscent antibody test.
 Two smears are to be made at the two end of the slides with a
sterile cotton swab soaked in saliva from the under surface of
patient’s tongue.
 At least 4 slides are to be made and air dried before sending to
laboratory to carry the test.
 Fluorescent antibody technique (FAT) : presence of rabies
antigen by fat in corneal smears, smears from saliva confirm the
diagnosis of rabies.

11. Babesiosis In Bovine
Etiology : Babesia bigemina
Sample collected :
 Peripheral Blood Smears (ear vein)
 Citrated blood
coffee color urine

13. Common method of diagnosis

15. Babesia bigemina in a Giemsa stained blood smear from a bovine

16. Trypanosomiasis / Surra
Sample collected :
 Peripheral Blood Smears (ear vein)
 Citrated blood
Common method of diagnosis
• Giemsa staining/

17. Theileriosis
Etiology : Theileria annulata
Sample collected :
 Peripheral Blood Smears (ear vein)
 Citrated blood
 Smear from enlarged lymphnode
Common method of diagnosis
• Giemsa staining/

20. A dog with severe demodectic mange
Demodicosis
Etiology : Demodex canis
Sample collected : small pinch of skin with
hair
Skin scraping Procedure:
1. Select the most active lesion i.e. periphery
lesion.
2. Clean the lesion with spirit.
3. Take a scalp and dip its blade in mineral oil i,e.
liquid paraffin.
4. Press the lesion between thumb and index
finger.
5. Hold the scalp blade with thumb and finger
firmly and scrape the skin deeply till little blood
oozes out.
6. Collect the scraping in petri dish / watch glass.

22. Deep Skin Scraping

23. Cigar shape Demodex canis under microscope

24. Scabies
Scabies in Dogs
Etiology : Sarcoptes scabiei mite
Sample collected : small pinch of skin with hair

27. Canine Distemper
 Sample Collected
 Blood
 Nasal Discharge Or Occular Discharge
 Conjunctival Swab
 Feaces
 Urine
 Diagnosis/Clinical Pathology
1. The TLC Will Vary, Depending Upon The Stage Of Disease
 In The Early Acute Stage, There Is A Leukopenia.
 Leukocytosis Is Present In Later Stages In About 20% Of The
Cases.
2. A Shift To The Left Of Neutrophilia Is Associated With The
Leukocytosis And Sometimes With Leukopenia.
3. Moderate Anemia May Be Seen In The Terminal Stages.

28. Other Methods
1. Canine Distemper Test Kid

29. Pyometra In Bitch
Definition : Accumulation of pus in uterus.
Sample collected
 Blood
 Vaginal discharge
Diagnosis / cinical pathology
 There is usually marked leukocytosis (20,000 to 100,000 or
more),with the average TLC being 50,000/ mocroliter or
over.
 There is marked is a marked neutrophilic shift to left.
a. The immature neutrophils often have a light blue staining
cytoplasm with toxic granulationand may contain Dohle bodies.
 The PCV is usually within normal limits, but
ocacasionally there may be a nonregenerative anemia or a
hemoconcentration.

32. Fasicioliasis
Sample collected
 Blood
 Feaces
 Diagnosis / clinical pathology
 In acute fasicioliasis there is a severe normochromic anaemia,
esinophilia, and severe hypoalbuminemia, serum enzyme are
elevated , increase in aspartate aminotransferase can results
(wyekoff and bradley, 1985)
 In subacute and chronic disease weight loss is associated with
a severe hypochromc, macrocytic anaemia, hypoalbuminemia
and hyperglobulinemia.

33. Milk Fever In Bovine
Definition : It is a metabolic disease of adult females
occurring most commonly about the time of parturition and
characterized by hypocalcemia, severe muscular weakness,
sternal and lateral recumbency , circulatory collapse and
depression of consciouness.
Sample collected
 Blood, milk
1. Haematology :
I. Increased PCV and Hb – due to dehydration
II. DLC count is suggestive of neutrophilia, eosinopenia and
lymphopenia which ocurs due to stress resulting from
adrenal cortical hyperactivity.

34. 2. Blood Biochemistry:
I. Serum Ca level is decreases to 5 mg % (Normal: 8-12
mg%).
II. Serum Pi level are usually decreased to 2-3 mg% (Normal:
4-6 mg%).
III. Serum Mg level are usually increased to 4-5 mg%
(Normal 2-3 mg%).
IV. Serum glucose level are usually decreased (Normal : 40-50
Mg %).
V. Ca : P ratio decreases to 1 : 1 (Normal : 2:1).
VI. Ca : Mg ratio decreases to 2 : 1 (Normal : 4 : 1).
3. Urine Analysis:
 Sulkowitch test is Negative.

35. Ketosis In Bovine
 Definitions: It is metabolic disease of high yielding animals
characterised by – hypglycaemia, ketonaemia and ketonuria.
 Sample collected
• Urine, Blood, Milk
 Diagnosis/ Clinical pathology
 Hypoglycaemia (< 40 Mg %) ,Ketonemia (>20 Mg% ) And Ketonuria.
I. Haematology
 DLC indicates neutropenia (10 %)
 Lymphocytosis (60-80%)
 Esinophilia (15-40%).
II. Urinanlysis
 urine sample is positive for ketones bodies.
III. Milk examination
 Milk Sample Is Positive For Ketone Bodies.

36. Rothera Test ( For Ketone Bodies)
Principle
Acetoacetic acid and acetone react with alkaline solution of sodium
nitroprusside to form a purple colored complex. This method can
detect above 1-5 mg/dl of acetoacetic acid and 10-20 mg/dl of
acetone. Beta-hydroxybutyrate is not detected.
Procedure
2 ml urine + Rothera’s reagent – deep purple color ring formed.
Rothera reagent – ammonium sulphate + sodium nitroprusside
(100:1).
Observations and Results
Immediate formation of purple permanganate colored ring at the
interface: Ketone bodies present (Positive)
No formation of purple permanganate colored ring at the interface:
ketone bodies absent (Negative)
Grade the result according to intensity of the formation of colored
ring as Trace, +, ++, +++ or ++++.

38. Hypomagnesium Tetany
Definition : it is highly fatal metabolic disease of
ruminents characterized by Hypomagnesia.
Sample collection
 Blood or serum
Diagnosis/ Clinical Pathology
 Serum magnesium : reduced to 0.5 mg % ( normal 1.7 – 3
mg %)
 Serum calcium : reduced to 5-8 mg% ( normal 8-12 mg %).
 Serum P : may or may not be reduced.
 Serum K : high (hyperkalaemia)

39. Acidic Indigestion In Ruminents
Definition : it is an acute ruminal dysfunction caused by
ingestion of large amounts of CHO rich feeds and clinically
characterized by severe toxaemia, dehydration, ruminal
stasis, weakness, recumbency and high mortality.
Sample collected
 Ruminal Fluid
 Urine
Serum
Diagnosis/clinical pathology
a) Ruminal fluid :
 pH – 5.5 to 6.5 mild acidosis
4.5 to 5.5 moderate acidosis
4.0 to 4.5 severe acidosis

40.  Rumimal fluid : colour – milky grey
odour – sour
 Absence of ruminal protozoa
 Bacteria : gram negative replaced by gram positive bacteria.
(normally Gram negative are predominent)
b. Urine analysis :
 ph – acidic/lowered(5.0)
 Proteinuria
c. Haematology – high PCV (increased 50-60%)
d. Serum biochemistry : increase BUN due to renal failure.
 Increased blood lactate
 Decreased bicarbonate.
 Mild hypocalcaemia – due to temporary malabsortion.

41. Alkaline Indigestion In Ruminents
 Definition : it is caused by ingestion of protein rich feeds and
characterized by anorexia, depression, alkaline rumen pH and
atony of rumen.
 Sample collected
 Ruminal Fluid
 Urine, Serum
 Diagnosis/clinical pathology
 Rumen Fluid
 Ph > 7.5 (alkaline)
 Colour – dark brown
 Odour - Ammoniacal
 Blood – increase BUN (>24 mg /dl)

42. Cyanide/Hcn Poisoning In Ruminents
 Sample collected :
 Oxalated blood
 Urine
 Suspected feed and water
 Rumen fluid
 Diagnostic by Picrate Paper Test
Reagent : sodium bicarbonate 5 g + picric acid 0.5 g + DW to
make 100 ml. Dip Paper Strips And Dry In Cool Place.
 Procedure
1. Put few drops of rumen fluid on picrate paper.
2. Change of colour from yellow to brown / red indicates
presence of cyanide.

43. Nitrate / Nitrite Poisoning in Ruminents
 Sample collected :
 Oxalated blood
 Urine
 Suspected feed and water
 Rumen fluid
 Diagnostic by
a. Clinical Pathology - Total Hb and erythrocyte concentrations
increase in proportion to blood methmoglobin levels.
b. Diphenylamine Blue Test
DPB Reagent : 0.5 g Diphenylamine +20 ml DW + Conc. H2SO4
to make up 100 ml.
Procedure
1. Put few drops of plasma/urine/rumen fluid on glass slide.
2. Put 1-2 drop of DPB reagent by side of same and allow to mix.
3. Develpoment of blue colour indicates positive test.

44. Diabetes mellitus in Dogs
Definition – it is a chronic complex metabolic diorder
caused by insulin insufficiency / deficiency or impaired
insulin action and characterized by glucose intolerance,
persistent hyperglycemia and glycosuria
Sample collected
• Blood (in sodium fluoride and thymol 10:1)
• Urine
Diagnosis
 Fructosamine
 is a compound that is formed when glucose combines with protein. This
test measures the total amount of fructosamine (glycated protein) in the
blood. Glucose molecules will permanently combine with proteins in the
blood in a process called glycation.

45.  Glycosylated haemoglobin
 Glycosylated haemoglobin is produced by the non-enzymatic,
irreversible binding of glucose to haemoglobin in erythrocytes.
 As the blood glucose concentration increases, the rate of
hemoglobin glycosylation also increases.
 The specific fraction of glycosylated hemoglobin that is
measured is HbA1c.
 Glycosylated (glycated) hemoglobin concentration can be used
as a screening test for diabetes mellitus, as well as for the
monitoring of glycemic control in treated diabetic animals.
 Glycosylated hemoglobin determination also is useful in long
term monitoring of diabetic patients over the previous 2-3
months.

46.  Blood Glucose Monitoring
 Collecting a drop of blood from the ear (pinna) or a carpal pad or a
footpad and analyzing this using a hand-held blood glucose meter.
 small test strip with the drop of blood on it is by inserted into a small
machine (hand-held blood glucose meter), which reads the strip and
shows the blood glucose level in a digital display window.
 he drop of blood can be produced using a sterile needle or a special
lancet (razor sharp device to puncture the skin).
Blood biochemistry
 Normal in dogs – 80 to 120 mg/dl.
 Repeated fasting blood glucose > 150 mg/dl
 Post prandial blood glucose >200 mg/dl
 The renal threshold for glucose is a blood glucose concentration of
around: In dogs: 10 mmol/l (180 mg/dl)
 If the blood glucose level exceeds this threshold, glucose is excreted
in the urine.

48. Urine analysis
Benedic test (for Glucose)
Anhydrous sodium carbonate = 100 gm
Sodium citrate – 173 gm
Copper(II) sulfate pentahydrate = 17.3
gm in 1 litre of solution
Procedure
Approximately 1 ml of sample is
placed into a clean test tube.
2 ml (10 drops) of Benedict’s
reagent (CuSO4) is placed in the
test tube.
The solution is then heated in a
boiling water bath for 3-5
minutes.
Observe for color change in the
solution of test tubes or
precipitate formation.
 Positive Benedict’s
Test: Formation of a reddish
precipitate within three
minutes. Reducing sugars
present. Example: Glucose

Negative Benedict’s Test: No
color change (Remains Blue).
Reducing sugars absent.

49. Nephritis
Definition : it means inflammation of kidney.
 Sample collected
 Blood
 Urine
 Diagnosis/clinical pathology
 Blood biochemistry
 BUN : increase (normal : 6-27 mg/dl)
 Serum creatinine - increase ( normal : 1-2 mg/dl)
 Serum total protein – decrease ( 5.7 - 8.1 gm/dl)
 Urinalysis :
 Acute – high specific gravity and presence of RBC , WBC, or
epithelial casts on microscopic examination.
 Chronic – low specific gravity with less cellular deposits.
 Proteinnuria – it is cardinal sign of glomerulonephritis. (Robert
test positive).

50. Microscopic Examination Of Urine

51. Traumatic Reticulo-Peritonitis (TRP)
Definition : it is caused by sharp pointed foreign bodies and
characterized by anorexia, mild fever, ruminal stasis and
subacute abdominal pain.
Sample collected
 Blood
Diagnosis / clinical pathology
 The TLC is usually increased early in the course of the
disease but later may drop rapidly.
 A neutrophilia with a marked shift to left is characterstic.
 High eosinophil counts are occasionally seen in some
chronic cases.

52. Neutrophils Shift To Left