SYED MUHAMMAD RAZA

SYED MUHAMMAD RAZA

[email protected]

Pir Mehr Ali Shah - Arid Agriculture University Rawalpindi (PMAS-AAUR)

Formalin and fumes can damage blood smear, therefore do not ship blood smears with formalinfixed tissue
samples.

Crenation, Punched-out cells, and Smudge cells are usually artifacts caused by errors during preparation of blood
films.

Bovine RBCs are smaller and take longer to pack down, so it is necessary to run the centrifuge longer for cattle
than for cats.

Lipemia is lipedemic serum (milk-colored, fatty) that usually happens after a recent meal. It affects many
biochemical tests.

Hyperlipidemia (high blood lipids) can be caused by sudden metabolic changes (pregnancy, stress) or decrease
in feed quality in horses. If the serum is left sitting too long, then the RBC bicarbonate and glucose can decrease,
and phosphorus and potassium can increase.

Packed cell volume (PCV): is another name for a hematocrit. Is given in percentage. The hemoglobin (Hb, g/dl)
correspond to 1/3 of the PCV. So, if an animal has 30% PCV, it has 10g/dl of Hb.

Red Blood Cells (RBC): Hemosiderin (and ferritin) are a storage form of iron.

Tubes Chemical Important points

Purple top EDTA It lyses RBC of birds and reptiles. EDTA is a calcium total chelate, therefore it
inhibits Ca from activating coagulation factors and platelet aggregation. It
does not stain the cells and doesn’t interfere with blood cell morphology.
Green Top heparin Used for RBC of birds and reptiles. Inhibits thrombin that converts fibrinogen
in fibrin.
It binds to antithrombin which subsequently accelerates the interaction
between antithrombin and activated factor X.
Blue top Sodium Citrate Reversible calcium chelate. PT & APTT
Red Top coagulation Serum chemistry
accelerator
Yellow top separating gel
gray top sodium fluoride Inhibit glucose breakdown, so it is used to measure glucose and lactate.

BLOOD SMEAR
Body Cells are densely packed. It is the are closest to where blood drop is placed.
Counting area Morphologically evaluated & counted
Feathered edge Examine large abnormal cells, platelet clump, microfilaria

TOTAL BLOOD VOLUME

Animals 7-9 %
Horse 8%
Cats 6.5 %
Life Spans
Red Blood Cells (RBCs)

• Dog 110 Days

• Horse 145
• Cat 70 Days
• Cow 160 Days
• Pig 86 Days
• sheep 150 Days

Neutrophils

Lifespan in all species is less than 24 hours.

• Dog 5-9 hours

• Cow 16 hours
• Horse 10-11 hours

Platelets

5-7 days circulating life span

Birds and reptiles have nucleated RBC.

Camelids (llamas, camels, and alpacas) have ellipsoid RBC without a central parlor.

Increase the angle between slides of anemic animals.

Romanowsky stains include Wright’s and DiffQuik that are used to stain blood smears.

Reticulocytes (retics): In case of anemia small animals will release reticulocyte and metarubricyte. Ruminants
rarely release reticulocyte. Horses NEVER release reticulocyte and metarubricyte. Reticulocytes can be punctate
(mature, single dot) or aggregate (immature strand). In felines punctate retics can last for 7-10 days, therefore
we only count aggregate retics in felines. In other species, you count both the punctate and aggregate. Use
Methylene blue to look for reticulocyte. If you use Wright’s stain the reticulocytes will appear polychromatic.

New Methylene blue (NMB) is used to stain blood smear to look for reticulocytes.

Anisocytosis: Different sizes in between RBC of the same blood smear. Microcytosis (small cells) is sometimes
seen in dogs with congenital liver problems like portosystemic shunts.

Polychromasia: different color in between RBC of the same blood smear. It indicates regenerative anemia.

Poikilocytosis: difference in shape and size between RBC that is physiological in goats.

RBC index are blood tests that provide information about the hemoglobin content and size of red blood cells.
They are calculated from RBC measures like erythrocyte count, packed cell volume and hemoglobin
concentrations. MCHC is the most accurate RBC index because is calculated based on packet cell volume. MCH
is the least accurate because it is calculated based on RBC count.

MCHC = hb/PCV MCH = hb/ RBC MCV = PCV/ RBC

Mean Corpuscular Hemoglobin Concentration (MCHC): color of the RBC and is recorded in g/dL. Reticulocytes
have 40% less hemoglobin and so they are less pigmented.

Mean Corpuscular Volume (MCV): size of the RBC that is recorded in femtoliters (fL). Reticulocytes are bigger;
therefore, a macrocytic anemia is regenerative. Normocytic tends to be non-regenerative. Microcytic is a non-
regenerative anemia.

Anemia: 3 causes: Loss, destruction or decrease production, so Hemorrhage, Hemolysis and Decrease in
production. After a severe hemorrhage, you can detect bone marrow response in 2 to 3 days, but maximum
response takes around 1 week. Lack of iron and complex B vitamins will result in a microcytic and hypochromic
anemia that is nonregenerative.

Horses that suffered acute blood loss do not have an alteration in PCV and protein in the first 12 hours. Shifts of
extracellular fluid usually takes 12 to 24 hours, and then you have a decrease in PCV and plasma protein.

Polycythemia: Increase in the RBC. Secondary polycythemia is when RBC are increased due to an underlying
cause such as dehydration and splenic contraction. Polycythemia Vera is when you have a disease that makes
the bone marrow to produce more RBC.

Rouleau Formation: piling of erythrocytes that is physiological in felines and horses. It is present during
inflammatory circumstances.

Target Cell: Bad distribution of hemoglobin in RBC due to hepatopathy, hemolytic anemia and iron deficiency.

Spherocyte: is a ball-shaped RBC that was damaged, but it can be caused by auto-hemolytic anemia.

Metarubricyte: nucleated RBCs

Schistocytes: are produced by mechanical fragmentation of erythrocytes on intravascular fibrin strands. Most
commonly a result of DIC, heartworm, Hemangiosarcoma and glomerulonephritis.

Basophilic stippling is the aggregation of ribosomes in the cytoplasm of a RBC, giving it a basophil look. You can
see in animals suffering from lead poisoning.

Acanthocytes: RBC with membrane abnormality that causes numerous, irregularly spaced projections. They
occur due to fragmentation injury, liver disease, DIC or congenital defects. It is a normal finding in young
ruminants (calves and kids with less than 3 months of age). However, it should be differentiated from crenation.

Crenation: is an artifact during making of blood smear that results in RBCs with rounded regularly placed
projections around cell membrane. It occurs if the film is dried too slowly or at high temperature.

Smudge cells are cells that have been damaged when excessive pressure is used during the process of creating
a blood smear.

Inclusion Bodies
Howell Jolly Heinz bodies
indicative of regenerative anemia, common in can be seen in ghost cells after the damaged Hb is
felines. denatured. It is also evidence of oxidant induced
hemolytic anemia caused by acetaminophen in cats
and red maple in horses. Can be seen in hepatic
disease.

Iron Deficiency: Microcytic Hypochromic. Non regenerative Anemia

Non-Regenerative Anemia: Reticulocytes count below 60000/ul

Regenerative Anemia in Equines:

The only accurate way to gauge response to anemia in or horse is with Bone marrow examination or Serial CBC
taken daily fox many days.

Hypercellular bone marrow with a low myloid /erythroid ratio ↑ MCV is consistent in horses with regeneration
anemia.

Most types of anemia in horses are, Normocytic, Normochromic

Echinocyte: Rattlesnake Bite, Hemangiosarcoma, Live diseases

Regenerative anemia will be seen with both Heinz body and immune mediated hemolytic anemia.
Leukocytosis means an increase in WBC number.
Leukopenia means decrease in number of WBC
(leukocytes).

Polymorphonuclear cells Family (PMNs): include

the eosinophils, neutrophils, and basophils. All WBC
are phagocytic, except thrombocytes (platelets) and
lymphocytes.

Neutrophils: Neutrophilia is the increase in neutrophils due to bacterial infection (commonly), recent systemic
inflammatory response, tissue necrosis, autoimmune disease, and corticoid usage for long period of time.
Neutropenia means low levels of neutrophils. Healthy neutrophils may have barr bodies.

Eosinophils: stain red. Eosinophilia is the increase in eosinophils and happens due to tissue lesion (fleas and
ticks), allergic disorders and parasitism. Eosinopenia means low levels of eosinophils.

Basophils: stain dark blue. It has a similar function to mast cells.

Monocytes: when they migrate to the tissue, they are called macrophages. Play a significant role in intracellular
pathogens such as fungi, virus, and some bacteria (brucellosis and mycobacterium). Monocytosis is an increase
in the number of monocytes and monocytopenia is a decrease in monocytes. Macrophages in the liver are
known as Kupffer cells, in the bones are osteoclasts, in the connective tissue histiocytes and in the nervous tissue
microglial cells.

Lymphocytes: responsible for immunologic activity. It is not possible to differentiate a T and a B lymphocyte in
a blood smear. Lymphocytosis is a high lymphocyte count and Lymphopenia is low levels of lymphocytes. High
nuclear/ cytoplasm ratio, chromatin is coarse, clumped & dark staining and sky-blue cytoplasm.

Hyper segmented Neutrophil: neutrophils with more than 5 segmentations. It is an indicative of vitamin
deficiency (B9 and B12), chronic inflammation and megaloblastic anemia. Exogenous or endogenous steroid can
cause hyper segmented neutrophils.

Left shift: indicates that immature (no segmented) neutrophils have been released from the bone marrow into
circulation prior to maturation to segmented cells. These immature neutrophils are also known as band cells.
(U- shaped)

Toxic Change: are indicative of inflammation. They will include Dohle bodies (pale blue irregularly shaped
cytoplasmic inclusions), increased cytoplasmic basophilia, and foamy/vacuolated cytoplasm. Toxic change
indicates accelerated production of neutrophils by the bone marrow.

Stress Leukogram: is characterized by neutrophilia, lymphopenia, eosinopenia, and potentially Monocytosis. It

occurs primarily in dogs in response to stress, bacterial infection and hyperadrenocorticism. The term stress
denotes the presence of increased cortisol released from the adrenal gland secondary to severe disease (e.g.,
diabetic ketoacidosis, renal failure), high body temperature, pain, dehydration, or hyperadrenocorticism.

Inflammatory Leukogram: may be characterized by neutrophilia, or in some instances neutropenia, with or

without a left shift or toxic change.

Fibrinogen is markedly increased (hyperfibrinogenemia) in most cases of chronic inflammation.

Physiological leukogram: it is from an epinephrine mediated excitation response and is commonly seen in cats
who get excited upon entering a veterinary clinic. Mild neutrophilia, normal to increased lymphocytes, no
monocytosis.
Platelet plug formation occurs within 5 minutes.

Platelets are produced by megakaryocytes. 1/3 of platelets are stored in the spleen, therefore animals that had
a splenectomy will have higher concentration of platelets in the blood.

Coagulation cascade: The extrinsic pathway (factor 7), intrinsic pathway (factor 12, 11, 9 and 8) and common
pathway (factor 10, 5, 2 and 1) together create thrombin that transform fibrinogen into fibrin.

Feline platelets clump easily, so take that in consideration while counting.

Thrombocytosis: increase in number of platelets. Usually due to dehydration, but it can be pathological such as
in FeLV cases.

Thrombocytopenia: decrease in platelets number. Occurs due to: Destruction, Consumption and decrease
production. Consumption can happen due to DIC (Disseminated Intravascular Coagulation), neoplasia or
splenomegaly. Destruction can be due to parasites (Anaplasma) and immune mediated. Decrease platelet
function happens due to uremia, hepatic disease (no production of coagulating factors), antibiotics such as
penicillin and cefazolin, NAIES and calcium blocking drugs (some cardiac drugs and barbiturates).

Dogs and cats usually do not have spontaneous bleeding until number of platelet count is below 30000/ul.

Buccal mucosal bleeding time (BMBT): Evaluate primary hemostasis, therefore it only evaluates platelet plug
formation and NOT fibrin formation.

Prothrombin time (PT): measures the overall speed of the extrinsic and common pathways.

Activated Partial thromboplastin time (aPTT): measures the overall speed at which blood clots by means of two
consecutive series of biochemical reactions known as the intrinsic (contact activation pathway) and common
coagulation pathways. It is used in conjunction with another measure of how quickly blood clotting takes place
called the prothrombin time (PT).

Thrombin time (TT): is a blood test that measures the time it takes for a clot to form in the plasma of a blood
sample containing anticoagulant, after an excess of thrombin has been added. It evaluates how long it takes to
convert fibrinogen to fibrin.

Fibrin degradation products (FDP): It evaluates how long it takes to dissolve the clot into plasmin.

Protein Induced by Vitamin K Antagonist (PIVKA): It measures the precursors to the coagulation proteins that
accumulate when vitamin K is absent. Is the most sensitive test for anticoagulant rodenticide toxicity.

DIC: think diseases with H: Heartworm, Heart failure, Hemolytic anemia, Hemangiosarcoma, Hemorrhagic
gastroenteritis and Hepatic disease. Other causes include: Gastric Dilatation Volvulus and pancreatitis.

Blood Transfusion
Major crossmatch Minor crossmatch
determines the presence of preexisting isoantibodies determines the presence of preexisting isoantibodies
in recipient plasma that could produce adverse in donor plasma. This test is important for animals
reactions. that have naturally occurring isoantibodies such as
cats and some dog breeds (German shepherd), in
patients that have already had a transfusion and in
multiparous mares.

Hemophilia A

It is transmitted by an X-linked pattern of inheritance (recessive trait) so females Less likely affected.

Females with the gene of disease are usually unaffected carriers unless they Leve the offspring of an affected
male and a carrier female.

Affected animals usually have less than lot of factor VIII s prolonged ACT & APIT. Treatment of choice to stop a
bleeding event is fresh plasma or fresh frozen plasma.

Von willebrand’s

(73 %) Doberman. Autosomal trait.

Rx: Desmopressin acetate (DDAVP)

The enzymes AST, ALT, SDH and LDH are indicative of hepatocellular leakage.

AST (aspartate aminotransferase): indicative of rhabdomyolysis (skeletal muscle of large animals) or

hepatocellular leakage (small animals). Great marker for bird hepatic disease.

ALT (Alanine transaminase): only exists inside of hepatocytes and is an indicator of liver damage in small
animals.

ALP or AP (Alkaline Phosphate): It has 5 isoenzymes in dogs. Increase with excess osteoblastic activity, therefore
may be increased in growing puppies. It also increased in pregnant animals, high stress levels, cholestasis,
intestinal disease, and steroid therapy.

Liver-ALP
Placenta-ALP
Bone-ALP
Leukocyte-ALP
Corticosteroid-ALP (Occurs in dogs only)

GGT: inducible enzyme that increase in cholestasis but is present in other tissues. High in neonates due to
colostrum absorption and in animals that are treat with anticonvulsants and glucocorticoids. Go to enzyme for
hepatic disease in horses. However, in large animals is also present in the liver, pancreas, and kidneys.

SDH (Sorbital dehydrogenase): very useful for evaluation of acute hepatitis in large animals.

CK (Creatine Kinase): indicative of rhabdomyolysis. It will also be increased in horses with ionophore toxicity.

LDH (Lactate dehydrogenase): indicated hepatocellular leakage enzyme evaluates in large animals but can be
increased in muscle injury.

Bile acids: to evaluate liver function and possible shunt.

Bilirubin increases in hemolytic diseases. Cholestasis may increase bilirubin.

Serum Albumin: more than half of total blood protein. Increase value may be due to dehydration. Decrease
values can be by decrease production (hepatopathy), low dietary protein and loss of protein (enteropathy or
renal disease).

Potassium (K): hypokalemic myopathy caused paralysis and the neck has a ventroflexion and is stiff.
Hyperkalemia is cardiotoxic and causes bradycardia, large, spiked T waves, flat or absent P waves, wider QRS
complex and S-T segments depression.

Rx: Ca gluconate, Insulin, Dextrose, Sodium bicarbonate

Calcium (Ca): Hypercalcemia is usually idiopathic. Because of renal disease (hypocalcemia), eclampsia
(hypocalcemia), phosphate enemas (hypocalcemia). Hypercalcemia in Neoplasia (lymphoma).

Pancreatic Specific lipase: great to diagnose pancreatitis, but it can give a false positive with renal lesions.

Amylase: Hyperamylasemia is often seen in dogs with exocrine pancreatitis.

Trypsin-like immunoreactivity: tests pancreatic function.

Chylothorax is an obstruction of the thoracic duct that leads to accumulation of chyle in the thorax. It is a
modified transudate, white, pink in colour typically opaque. It is the accumulation of chyle (triglyceride) fluid
resulting from lymphatic drainage of GIT into pleural cavity. Normally chyle returns to the cranial vena cava via
thoracic duct and it can be secondary to thoracic duct trauma, CHF, or a cranial mediastinal mass or idiopathic.
Anion Gap (AG): Positive cations (Na++K+) minus negative anions (Total CO2 (HCO3) - + Cl-). Total CO2 - can be
replaced by bicarbonate (HCO3 -). Anion gap increases with diabetic ketoacidosis, renal insufficiency, Ethylene
glycol toxicity, grain overload and strenuous exercise.

Metabolic acidosis: caused by decreased concentration of bicarbonate (HCO3 -). It can happen due to loss of
bicarbonate by diarrhea or saliva loss (ruminants) or Titration of Bicarbonate during grain overload, diarrhea,
ketosis, diabetic ketoacidosis, renal disease, and some toxins such as ethylene glycol.

Total bicarbonate deficit is calculated: Total bicarbonate deficit (in mEq) = base deficit ×0.3×Body weight (in
kg). Typically, 1/3 to ½ of the calculated deficit is administered over 5-10 min and then the remaining is
administered over following 12-24 hours.

Normal base deficit of dog = -5 to 0 mmol/L.

Metabolic Alkalosis: caused by decrease concentration of HCl. (Vomiting, displaced abomasum)

Respiratory alkalosis and acidosis are caused by changes in pCO2. Hyperventilation and reduction in pCO2
caused respiratory alkalosis, while hypoventilation causes an increase in pCO2 and respiratory acidosis.

Urinalysis should be done as soon as possible. Use a drop or 3 of re-suspended sediment after removal of
supernatant from centrifuged urine for examination. Perform tests within 30 minutes.

Herbivores have alkaline urine and carnivores have normally acidic urine. Anorexic animals have acidic urine.
Cats & goats have pungent urine. Horses have cloudy urine.

Cystocentesis is contraindicated in animals with bleeding disorders, ascites, peritonitis, dyspnea, or pain that
would cause distress in dorsal recumbency.

Glomerular filtration rate (GFR) is measured ml/min/kg or by ml/min/square meter. Azotemia: excess of
nitrogenous compounds in the blood. Pre-renal azotemia signs include increase BUN and dehydration. Uremia
is the clinical manifestation of azotemia. Urinary specific gravity (SG) evaluates the kidney capacity of
concentrating urine. Classified in hyposthenuria (less or equal to 1.007), isosthenuria (1.008-1.012) and
hypersthenuria (>1.013). Isosthenuria means the urine osmolality is the same as that of plasma. Bilirubin gives
the urine a brownish-greenish color and it usually precedes icteric signs.

Blood Urea Nitrogen (BUN): suffers influence of hepatic production, diet, gastrointestinal hemorrhage and is
reabsorbed by renal tubules.

Creatinine: constant daily production, suffers small dietary influence, excreted only by glomerular filtration, and
does not suffer reabsorption or secretion. However, it can be bigger in animals with high muscle mass. Creatinine
is more sensitive indicator for acute renal failure than BUN in large animals.

Leukocytes: less than 5 leukocytes per high power field in the urine is normal, more than that is a sign of urinary
tract infection and can be classified as pyuria. Degenerated neutrophils in the urine are known as pyocytes.
 URINARY CRYSTALS

Struvite crystals: These are magnesium ammonium phosphate crystals, also known as triple phosphate crystals
or coffin lids. Often found in alkaline urine.

Calcium oxalate Dihydrate crystals: look like square diamonds and can be found in normal urine.

Calcium oxalate Monohydrate crystals: look like lozenges and sometimes can be seen in antifreeze toxicity
(ethylene glycol)

Calcium carbonate: wheel-like shape and often appears in healthy horse urine.

Makes urine of horse and rabbit cloudy.

Uric Acid crystals: found in Dalmatians as they lack a liver enzyme.

Promoted by high acidic diet & high purines diet which are metabolized to uric acid. Soy is high in purines.

Alkalizing and low purine diet is recommended.

Allopurinol therapy is done. It acts by inhibiting the enzyme xanthine oxidase, which metabolizes xanthine. It
works by stopping purine metabolism pathway at this point so uric acid will not be formed in high quantities.

It can cause xanthine crystals formation.

Amorphous phosphate: granular precipitate, found in alkaline urine.

Ammonium biurate crystals: Found in animals with portosystemic vascular anomaly (shunts)

Cystine: Form as a result of a genetic defect of the renal tubules and are almost exclusively seen only in males.
↓ Protein, Sodium restricted. Urine alkalizing diet & 2-MPG

Calcium oxalate
Most difficult and unlikely to dissolve.

Require Surgery, voiding hydropulsion, Lithotripsy

Dietary management: Avoid excess protein, calcium oxalate, vit-1), Sodium Provide adequate P. Mg. Vit. B6
Alkalizing diet to prevent new stones

Key Points
o Most common in Dogs, Cats → Struvite, Calcium oxalate
o All types of stones can be dissolved medically except Calcium oxalate.
o Use Alkalizing diet → Cystine, Urate crystals
o Use Acidifying diet → Struvite crystals
o Radiolucent (not visible on X-ray) → Cystine, Urate

Decrease protein diet for management of cystine, Struvite, urate, Canine struvite & to lesser degree calcium
oxalate.

Urine Alkalizer: Potassium citrate (preferred) & Oral Sodium bicarbonate

Casts: made of Tamm-Horsfall proteins! It may indicate tubular damage.

Hyaline casts if clear or semitransparent they are an indicative of mild glomerular leakage.
Cellular casts like sloughed epithelial cells or RBC with renal hemorrhage or WBC with
pyelonephritis.
Waxy casts Older version of granular casts and may indicate severe renal tubular
degeneration. They are wide, blunt, or squared ended and opaque.
Round cell tumors include Mast cell, Histiocytoma, Lymphoma, Plasmacytoma, Melanoma, basocellular tumors
and TVT.

Mast Cell Tumor think cytoplasmic granules

Histiocytoma occurs only in the dog and is the most common skin tumor in
this species. Occurs in young animals (50% are less than 2
years old). Rapidly growing, dome shaped, often ulcerated
lesions approximately 2 cm in diameter. Skin of the head is the
most common site. Do not metastasize and may
spontaneously regress.
Plasmacytoma Round, eccentric nuclei with dense chromatin; bi- and
trinucleate cells are common and characteristic. Image
bellow. Marked pleomorphism.

Transmissible Abundant cytoplasm contains distinct clear vacuoles.

Venereal Tumor
(TVT)
Can be cured in 90% or more cases with chemotherapy alone.

Lymphoma Originates in solid tissue (spleen, lymph nodes and thymus) Lymphoid
and bone marrow
Leukemia Originates from blood circulation and bone marrow. They Lymphoid + Myeloid
arise from myeloid cells (monocytes, basophils, eosinophils,
megakaryocytes and erythrocytic precursor cells).
Daily testing for the LH surge is the most accurate method of ovulation timing as LH surge becomes day 0.

Anestrus has a
predominance of
parabasal and
intermediate cells with
only a few neutrophils and
bacteria. The cellularity is
low (A).

Proestrus (early estrogen

influence): From early to
late proestrus, a gradual
shift from parabasal and
intermediate cells (small
then larger) and finally
superficial cells occurs (B).
Typically, red blood cells
are present in large
numbers (C)

Estrus think about corn

flakes! Look for more than
90% cornified superficial
cells with angular, sharp
edges and tiny pyknotic
nuclei or no nuclei (D, E).
Full cornification usually
coincides with receptivity.
The image in the side
shows cornification of cells
which is a sign that
estrogen is decreasing, and LH surge is imminent or just happened.

Diestrus (the luteal phase): Onset of diestrus is marked by a precipitous decline in the number of superficial cells
and reappearance of intermediate and parabasal cells within 1 to 2 days. Neutrophils are commonly observed
(F), and large numbers of bacteria are also often present (G).
Gram positive: Blue/purple. Has teichoic acids.

Gram Negative: Red. Has an outer membrane and LPS (endotoxemia).

Acid-fast staining is done primarily to detect mycobacteria, which causes bovine tuberculosis (mycobacterium
bovis) and Johne’s disease (Mycobacterium avium subspecies paratuberculosis).

Catalase test that evaluates the presence of catalase enzyme that breaks down hydrogen peroxide producing
water and oxygen.

Coombs test: Used for IMHA and neonatal isoerythrolysis. Direct Coombs test is used to detect the antibodies
or complement proteins that are bound to the surface of red blood cells and is used for autoimmune hemolytic
anemia. A blood sample is taken, and the RBCs are washed (removing the patient's own plasma) and then
incubated with anti-human globulin (Coombs reagent). If this produces agglutination of RBCs, the direct Coombs
test is positive. Indirect Coombs test detects antibodies against RBCs that are present unbound in the patient's
serum. In this case, serum is extracted from the blood sample taken from the patient. Then, the serum is
incubated with RBCs of known antigenicity and anti-human globulin is added. If agglutination occurs, the indirect
Coombs test is positive.

Immune mediated hemolytic anemia (IMHA)

Cocker Spaniels only breed, genetically predisposed to IMHA

Normocytic, Normochromic Regenerative Anemia, Agglutination, Spherocytes

Mild to moderate Bilirubinemia

Rx: immunosuppressive therapy (Prednisone, cyclosporine)

Blood transfusion if PCV is below 20%

Immune mediated Thrombocytopenia

Rx: immunosuppressive doses of Corticosteroids, Vincristine

Cyclophosphamide help to induce remission

Azathioprine helps to maintain → maintain remission, can be given long-term with less side effects

Coffin's Test:
Diagnostic test to confirm EIA. This test is an afar immunodiffusion to detect Serum antibodies against the
retrovirus causing ETA.

Effective at diagnosing infection in chronic asymptomatic carriers

In acute phase of infection, not enough antibodies are made

In foals, false-positive occur due to acquisition of antibodies in clostrum.

 Bacterial culture media examples
G+ Phenyl ethyl alcohol agar
G- MacConkey agar
Anaerobic Brucella blood agar
Salmonella Selenite broth
Staphylococcus Mannitol salt agar
Enterobacteriaceae Citrate test media
Show how an organism use Glucose, Sucrose, Lactose and Triple sugar ion slant agar
whether it produces Hydrogen sulfide

Dermatophyte test media (DTM) is used to identify fungal pathogens like ringworm. If it is a positive fungal
culture, the DTM will change color. Another technique is the toothbrush, used specially in cats, as you scrub the
hair and then tap it into the DTM media.

Direct smear technique is preferred for detecting some types of protozoa in feces because they do not survive
other techniques.

PCR: can not distinguish between dead & live organisms. It makes millions of copies of specific DNA section. It is
extremely sensitive & rapid (4 hours).

ELISA: is used in bench top SNAP test. Antigen of interest bind to specific antibody then a substrate is added to
see colour change.

Immunohistochemistry: is similar to ELIS but can be performed on tissue samples not the fluids.

Fungal culture media: DTM, Sabouraud

Fungal staining: PAS, gomori’s methenamine silver and Gridley’s

Rivalta’s test: distilled water mixed with a drop of acetic acid in a clear tube. Then a drop of effusion fluid is
carefully layered on top. If the effusion fluid does not dissolve into the solution or form into a ball shape you
have a positive reaction for Feline Infectious Peritonitis (FIP).

Antimullerian hormone assay: is used to diagnose ovarian remnant. It is also used to diagnose a cryptorchid
male.

Helicobacter spp is a spirochete bacteria present in almost 100% of dogs and cats, but Helicobacter pylori is
rarely found in cats and was not reported in dogs.

Proteus bacteria: does not form discrete colonies when grown on blood agar and tends to swarm, spreading all
over the culture media which may obscure or inhibit other colonies. Use a selective media, like phenylethyl
alcohol, to minimize the effect.

Salmonellosis: Confirmed by isolate from enlarged mesenteric lymph nodes.

Staphylococcus aureus: gram-positive, catalase positive, coccus. Part of the normal flora, especially in birds and
humans. S. pseudointermedius is main in dog.

Streptococci: gram-positive, catalase-negative, non-motile, round and ferment sugars.

Leucocytozoonosis: infect the lymphocytes of birds. Signs of the disease are nonspecific such as lethargy,
decreased production, and death. Treatment is clopidol in feed.

Treponema paraluiscuniculi: causes rabbit syphilis (vent disease) Venereal disease. Similar to hutch burn.
Neorickettsia risticii: causes equine monocytic eherlichiosis or Potomac horse fever. It is transmitted by
trematodes (flukes).

Babesia bovis is a red blood cell protozoon of cattle.

Anaplasma spp. is an obligate intraerythrocytic bacteria, a tick-borne in the order of rickettsials.

Babesia Anaplasma

Bacterial culture & sensitivity for methicillin resistance

Oxacillin is preferred for this.

Oxacillin resistance implies resistance to all penicillins, cephalosporins, imipenem, B. lactam, B. lactamase
inhibitors & Combinations.

Oxacillin is used because it is more stable a widely used.

Alcohol based Sanitizing Products

Effective at Killing many types of Give a Gave bacteria.

• Methicillin resistant staphylococcus aureus (MRSA)

• Tuberculosis
• E. coli
• HIV
• Influenza
• Hepatitis virus

Rabies virus is exception

Not effective against:

• Non-enveloped viruses (Parvovirus)

• Bacterial spores (Anthrax, C. difficile)
• Protozoon oocytes (Giardia, Isospora, Cryptosporidium)

Maintenance Hosts of Leptospira Serovars

• Canicola → Dogs
• Hardjo → Cattle
• Pomona → Swine, Opossum, Skunk, Racoon
• Icterohemorrhagiae → Rat
• Grippotyphosa → Mice, Squirrels Muskrats
 Tracheal wash with a few neutrophils and macrophages is normal. Too many is
abnormal and an indicative of respiratory infection. In addition, Simonsiella (in the
side image) is bacteria normally inhabit the pharynx and have a large and ladder-like
appearance. Presence of Simonsiella indicates bacterial contamination.

Schirmer’s tear test (STT) strip evaluated the amount of moisture in the eye. A
normal eye should have 18-25mm. Keratoconjunctivitis sicca is one disease that
Simonsiella causes dry eye, in cats STT would be around 10mm and in dogs it would be around
15mm.

Endometrial biopsy predicts the mare’s ability to carry a foal to term. The mare is categorized in I (80-90 %),
IIA (50-80 %), IIB (10-50 %) and III (10 %). III is the worst and the mare should not be used for breeding. Most
mares are category IIA. (Kenny scoring)

The mucin clot test (JOINT FLUID EVALUATIN) is a way to evaluate joint fluid viscosity. Normal joint fluid is
very viscous because it contains mucin. Joint fluid might be evaluated for signs of infectious, inflammatory, or
neoplastic arthritis in cases of lameness, joint swelling. Add equal volumes of 2.5% glacial acetic acid and joint
fluid and watch for formation of a tight white clot in the tube. If no good clot forms (i.e., it is not obvious or
soft), the mucin has been degraded by infectious agents or diluted by effusion in the joint.

Another subjective determination of joint fluid viscosity is the wooden stick applicator test. A wooden
applicator stick is placed into the joint fluid sample and slowly pulled out, forming a strand (normal strand is 2-
3 cm long).

Features of infected joints:

1. Total protein greater than 4.5 g/dl

2. Total nucleated count > 25000/ul
3. Polymorphonuclear cells 20000/ ul

Acid fast staining → Mycobacteria, Nocardia

KOH test → Fungal infections (Dermatophyte)

Congo Red stain → Amyloid fibrils & specific type of shigella

Silver staining → Identify proteins Type II collagen, fungal organisms

Zin Sulfate turbidity test → Failure of passive transfer

Synergistic Hemolysis inhibition test → Corynebacterium Pseudotuberculosis

Pulmonary Artery pressure Measurement to Predict High Mountain disease susceptibility

It is reliable above 6000 ft or animals above 12 months of age. BMPR-2 Gene is involved.

Readings less than 40 mmHg pressure are Satisfactory to purchase so the lowest PAR readings are the least
susceptible animals.

Procedure: Using local anesthesia a catheter is inserted into the jugular vein and Threaded into the Right Heart
and pulmonary artery where the pressure is taken while the animal is restrained in a squeeze chute.
The condition is caused by genetic predisposition to developing pulmonary hypertension a when Or tension is
reduced. The pulmonary hypertension leads to pulmonary artery hypertrophy, cor pulmonale and Right Heart
failure. Distended jugular vein, ventral edema, diarrhea, Pulmonary edema. Pleural effusion

Predisposing factors: Locoweeds Plants, Pregnancy, Cold weather

Fanconi syndrome: inherited disease in Basenjis. Renal tubular defects causing an abnormal loss of
electrolytes and solutes leading to decrease P, K, metabolic acidosis & glucosuria.

Myasthenia gravis: result of autoantibodies binding to the acetylcholine receptors, preventing

neurotransmission. Therefore, the animal cannot walk properly. Diagnosis with an acetylcholine receptor
antibody assay (best available) or a tensilon (edrophonium) test. A muscle biopsy is needed to quantitate
acetylcholine receptors for a definitive diagnosis of congenital myasthenia gravis.

Nosocomial infection: It is a hospital-acquired infection. Examples include parvovirus, kennel cough or

salmonella typhimurium (horses).

Diabetes Mellitus: Clinical finds include high blood sugar and glycosuria. It leads to diabetic cataracts in dogs,
but not in cats! It can also cause pancreatitis (more in dogs than in cats).

Diabetes insipidus: to differentiate diabetes Insipidus (DI) from psychogenic polydipsia (PPD) you need to do a
Water deprivation test and ADH response test.

Water deprivation test: due to medullary washout (loss of the gradient between renal medullary intersitium and
urine) you must deprive the animal from water before ADH test.

ADH response: to differentiate Central DI (lack of ADH due to a problem in the posterior pituitary gland or the
hypothalamus) or Nephrogenic DI (kidney not responding to ADH).

If USG after ONLY the urine concentration test is great than 1.025-30 think PPD or atypical hyperadrenocorticism.

If USG is less than 1.025, perform the ADH test. If the urine concentrates to greater than 1.025-30 think central
diabetes Insipidus. If not concentrate after BOTH tests, think nephrogenic diabetes insipidus.

Cushing’s (Hyperadrenocorticism): In dogs, usually toy breeds, clinical signs include obesity, increased
cholesterol, comedones, panting, increased ALP and sometimes you can see a mild increase in glucose and
decrease in BUN. In cats you see hyperglycemia, concurrent diabetes mellitus (80% of cases), increase
cholesterol and increase ALT. Diagnostic test include Low-dose dexamethasone suppression test (LDDST) and
ACTH stimulation test. LDDST is more sensitive (few false neg), while ACTH stimulation test can identify
iatrogenic causes. When you hear the 4 Ps (Panting, Potbelly, PU/PD, and polyphagia) think Cushing’s. Can use
mitotane as treatment.

Addison’s (Hypoadrenocorticism): Evident when Na:K ratio is less than 27! Look for low sodium, high potassium,
bradycardia, low Cl, high Ca, azotemia, increased PCV and anemia. There is a high incident of Addison’s in
Standard Poodles. Keep in mind that in felines a low Na:K ratio may be associated with other diseases and
definitive diagnosis is an ACTH stimulation test.

Hypothyroidism: can cause facial nerve paralysis. More common in dogs, usually large breeds, and leads to tail
alopecia and tragic facial expression. Other areas of bilateral alopecia are common and include the lateral trunk,
caudal thighs, ventral neck, and dorsum of the nose. Treatment includes thyroxin supplementation.

Hyperthyroidism it is the most common feline endocrine and geriatric disorder usually caused by a functional
thyroid adenoma (adenomatous hyperplasia). In roughly 70% of cases, both thyroids’ lobes are enlarged. Thyroid
carcinoma is the primary cause of hyperthyroidism in dogs, but it is rare in cats (1-2% of cases). Clinical sings
include thin, big appetite and hyperactive cat. Lab finding include high AP. Diagnostic test of choice is serum
total T4 (TT4). T3 suppression and TRH stimulation tests are useful to diagnose mild hyperthyroidism. If left
untreated hyperthyroidism can lead to hypertension. Palliative treatment includes Methimazole (block thyroid
hormone synthesis). Curative treatment includes surgical removal of the affected gland or radioactive iodine
therapy to destroy the gland.

Total body water: is 60% of body weight. Is divided into intracellular fluid (inside of cells), extracellular fluid
(outside of the cell) and third space fluid (pericardial, pleural, and peritoneal fluid). The extracellular fluid
compose has 1/3 of the body water and is divided into interstitial tissue fluid (16% of Body weight) and plasma
(4% of body weight). The intracellular fluid has 2/3 of the body water and compose around 40% of body weight.
Keep in mind that a 10% dehydration means 10% decrease of body weight in a non-obese animal.

Sensible losses are the fluid the animal loses by urination. (Hint: sensible losses make sense). Insensible losses
are fluid the animal loses that are hard to measure, like vomiting, diarrhea, and breathing.

Shock the most common type of shock in veterinary medicine is hypovolemic shock. Hypovolemic shock
happens when the total circulating blood volume decreases via loss of fluid such as hemorrhage, vomiting,
diarrhea, extreme sweating (horses) and more. Obstructive shock occurs when cardiac output is mechanically
impaired, like in GDV, cardiac tamponade and more. Cardiogenic shock is seen when the heart fails. Distributive
shock develops when blood volume is pooled in the peripheral circulation like in sepsis, anaphylaxis, heatstroke,
and snakebite.

Hypobiosis: is a stage of parasitic life with arrested development. (T. canis, Cyathostomes, Ostertagia)

Hyperthyroidism Mild increase in ALP & ALT in felines

Stress & Glucocorticoids Increased ALP in dogs only
Feline idiopathic hepatic lipidosis Increase bilirubin & ALP but normal GGT
Cholangiohepatic syndrome Increased GGT
Extra hepatic bile duct obstruction
Hyperadrenocorticism Increased cholesterol & ALP
Horse with acute colitis & Diarrhea Leukopenia and Hypoalbuminemia
Displaced abomasum Metabolic alkalosis
Multiple myeloma Hyperglobulinemia

Ruminal PH
Normal 6
High grain diet 5.5
High roughage diet 6.5
Anorexia 7.5
Prolonged anorexia 7-7.5
Carbohydrate overload 4.5
Urea overfeeding 8.5