Blood

Examination in
animals
Prepared by
Dr.Sherzad Ibrahim

Blood
Blood is a tissue consisting of cells within a fluid matrix. Blood creates an internal
environment which directly or indirectly baths all cells of the body and protects it from the
external environment.
Blood contains red blood cells (erythrocytes), white blood cells (leukocytes) and platelets
(thrombocytes). Figure 2.3 on page 10 of Voigt illustrates the cellular components of
blood. Red blood cells contain hemoglobin and are responsible for carrying oxygen from the
lungs to the cells throughout the body, as well as carbon dioxide from the tissues to the lungs
for excretion. White blood cells are either granulocytes (contain granules in the cytoplasm)
or agranulocytes (do not contain granules in the cytoplasm.) Granular leukocytes include
neutrophils, eosinophil's and basophils. Agranulocytes include lymphocytes and
monocytes. WBCs are a critical component of the immune system. Platelets are cell fragments
from large multinucleated cells (megakaryocytes). Platelets are important in blood clotting or
hemostasis.
The clear to pale-yellow fluid portion of blood is called plasma. Five to ten percent of the
plasma consists of proteins. The majority of the proteins are albumin, globulins and
fibrinogen. Albumins transport numerous substances in the blood and are the main
determinant of the osmotic pressure. Globulins (alpha, beta and gamma) are important in
transport and immunity. Fibrinogen is important in blood clotting and the inflammatory
cascade. If blood is allowed to clot, the clotting factors are removed from the plasma and
the remaining fluid portion of the blood is called serum.

the blood separates, under the influence of

gravity, into three layers). The bottom layer is
deep red in colour and is composed of red cells.
The top layer is clear and pale yellow. It is called
plasma and is composed of various salts and
proteins dissolved in water. In between is a
narrow layer called the buffy coat because of its
buff or yellowish white colour. The buffy coat is
composed mainly of cells of a variety of types,
collectively known as white cells. In addition
there are small cellular fragments, called
platelets, which have a role in blood clotting.

blood collection
The blood is collected from animal through puncture
of jugular vein in horse ,cattle,camel,sheep and goat
&from cephalic vein (recurrent tarsal vein)in dog and
cat
And from ear vein for isolation of blood parasite
with exception of tryponosoma(from J.V)

Functions of blood
The three main functions of blood are transportation, regulation and defense.
Many of these functions will be covered in detail in other units.
Oxygen and nutrients to cells in the body

Transports

Carbon dioxide and waste materials from cells in body

Hormones from glands to target organs

Body temperature

Regulates

Water balance
pH
Electrolytes
Phagocytosis of foreign invaders

Defense

Involved with immunity

Blood clotting

Organs associated with circulatory system

Many organs are associated with the circulatory system. They include

Heart:

Pumps blood throughout the body

Lungs:

Gas exchange: Oxygen and carbon dioxide

Liver:

Produces clotting factors and albumin

Removes waste material from blood

Spleen:

Blood storage
Removal of dead and damaged RBCs
WBC production in the lymphoid tissues of the
spleen

Bone
marrow:

RBC and WBC production

Blood Examination
A-Whole blood sample:
The blood sample is mixed with anticoagulant such as:heparine or E.D.T.A
It is used for hematological examination such as:
1. R.B.Cs parameter consist of : Red blood cell (erythrocyte)count ( RBC )
Packed Cell Volume ( P.C.V ) is the percentage of packed red cells in whole blood
Hematocrit (HCt) it is calculated from the RBC and MCV
Hemoglobin (Hb)
Mean Cell Volume (MCV) This is a measure of RBC size it represents the average volume of a red
cell and is expressed in femtolitres (fl). (10-15l).
Normal MCV = normocytosis means normal size
Increased MCV = macrocytosis indicates larger size and is usually caused by vit B12 and folic acid deficiencies
Decreased MCV = microcytosis i indicates smaller size due to iron deficiency etc and thus lower hemoglobin
concentration which may result from chronic haemorrhage or less commonly from dietary deficiency in young animals

Mean Corpuscular Haemoglobin (MCH) associated with hemoglobin concentration

Decreased MCH causes hypochromic anemia because of a deficiency of iron in the diet
Increased MCH may be caused during hemolysis where extracellular hemoglobin level increases

Mean Corpuscular Haemoglobin Concentration (MCHC) it describes the average

concentration of Haemoglobin per red cell
Normal MCHC = Normochromasia Lipaemia or icterus
Reduced MCHC = Hypochromasia Regenerative Anaemia and Iron deficiency or inefficient iron use

Blood Examination
2. W.B.Cs (leukocytic )count
3. differential leukocytic count
4. blood film

B-Serum sample:
The blood sample is collected without addition of anticoagulant
and then left to clot
Then centrifuged at 3000 r.p.m for 20 minutes
Take the clear serum and separate it in a clean plastic container
for biochemical examination
C. Blood film
a drop of fresh blood is placed in one corner end of slide and
spreader as smear with the help of another slide using it is thin
edger are an angle of 45(thin blood film)
b-dry the smear in air
c-fix the smear in methanol for 4-5 minutes or absolute methyl
alcohol for 1-2 minutes the process know fixation
d-dry in air
e-stain the smear with gimsa stain diluted (1:10)in distilled water
for 5 minutes
f-wash the slide with water
g-dry in air
h-examine under oil immersion lens
After staining, the structure and colour of red cells is enhanced
and the white cells and platelets to be recognized.

N.B:
*in case of trypanosomamake a thick blood film
***indication of blood film:
For detection of the presence of blood parasite such
as:babesia,thileria,anaplasma,tryponosoma and
filaria
For differential leukocytic count

RBC Erythrocytes

Erythrocytes from all mammals are anucleated, and most are in the shape of biconcave discs
called discocytes," The biconcave shape results in the central
pallor of erythrocytes observed in stained blood films.

Red cells are produced in the bone marrow and usually lose their nuclei when they are released
into the blood stream.

Color of RBC The presence of bluish-red erythrocytes in stained blood films is called
polychromasia. Polychromatophilic erythrocytes are reticulocytes that stain bluish red due to the
combined presence of hemoglobin (red staining) and individual ribosomes and polyribosomes
(blue staining).

WBC Leukocytes

Have nucleus, mitochondria, & amoeboid ability

Can squeeze through capillary walls (diapedesis)
Granular leukocytes help detoxify foreign substances & release heparin
Include eosinophils, basophils, & neutrophils

A Granular leukocytes are phagocytic & produce antibodies

Include lymphocytes & monocytes

Neutrophils (PMNs)

Multi-lobed nucleus with fine granules

Act as phagocytes at active sites of infection

Inflammation: Left Shift

increase in the numbers of immature neutrophils in the circulation
Eosinophil's

Large brick-red cytoplasmic granules

Found in response to allergies and parasitic worms

Causes of Eosinophilia
Hypersensitivity (allergy) e.g.. Flea bite hypersensitivity, asthma, pulmonary
infiltration with eosinophilia (PIE)
Parasitism (doesnt consistently cause an eosinophilia) for example intestinal
parasitism, aelurostrongylus, protozoa
Idiopathic eosinophilic diseases e.g. eosinophilic gastroenteritis, eosinophilic
myositis, idiopathic hypereosinophilic syndrome of cats, panosteitis
Infectious diseases e.g.. FIP
Neoplasia mast cell tumours, paraneoplastic eosinophilia, eosinophilic
leukaemia (rare)
Hypoadrenocortcism*

Eosinopenia
the eosinophil count falls within the normal range
Causes of Eosinopenia
Infection/inflammation
Adrenaline response acute stress, pain, excitement , exercise
Steroid response- chronic stress/illness, glucocorticoid administration
Basophils
Have histamine-containing granules
allergic and inflammatory responses and in the control of parasitic infections

Monocytes

Largest of the white blood cells

Function as macrophages
Important in fighting chronic infection

Causes of Monocytosis
Inflammation especially chronic or pyogranulomatous inflammation (TB/FIP)
Steroid response- chronic stress/illness, glucocorticoid administration,
hyperadrenocorticism
Monocytic or myelomonocytic leukaemia (monocytosis likely to be extreme)

Lymphocytes

Nucleus fills most of the cell

Play an important role in the immune response

Provide a specific immune response to infectious diseases.
There are 2 types: T-cells attacks cells bearing foreign antigens and antibody-coated cells; can help or suppress B cells
(part of cell-mediated immunity)
B-cells matures into a plasma cell, which secretes antibodies (humoral immunity)
They produce antibodies.

Causes of Lymphocytosis

Adrenaline response (as above more marked in cats)

Chronic inflammation infection or immune stimulation ( including post vaccinal)
Lymphoid leukaemia the lymphocytosis can be extreme especially with chronic
lymphocytic leukaemia
Hypoadrenocorticism (mild)

Causes of Lymphopenia

Acute viral infections

Steroid response (Lymphopenia is the most consistent of all the steroid induced
changes in leucocyte numbers)
Depletion of lymph e.g.. Loss into body cavity -chylothorax, or loss of afferent lymph
enteric pathology such as lymphoma or lymphangiectasia
Lymphoid hypoplasia e.g.. combined immunodeficiency in Basset Hounds,
chemotherapy, destruction of lymphoid tissues multicentric lymphoma

Platelets

Also called thrombocytes.

Derived from ruptured multinucleate cells (megakaryocytes)
Smallest of formed elements.

Are fragments of megakaryocytes.

Lack nuclei.
Have amoeboid movement.
Important in blood clotting:
Constitute most of the mass of the clot.

Release serotonin to reduce blood flow to area.

Secrete growth factors

Maintain the integrity of blood vessel wall.

Serum Examination
Tests Included in a Basic Test Panel

Test

What the Results may Mean

Total Protein

Increases due to dehydration or inflammation; may decrease

due to bleeding, malnutrition, or congestive heart failure

Albumin

Increases due to dehydration; may decrease due to bleeding,

congestive heart failure, or liver failure

Globulin

Calculated as the difference between total protein and albumin

Urea

Increases due to certain dietary excesses or deficiencies,

congestive heart failure, kidney failure, or a ruptured bladder;
decreases may be due to liver failure or low levels of dietary
protein

Creatinine

Glucose

ALT and ALP

Increases may be due to kidney disorders, muscle damage, or a

ruptured bladder
Increases may be caused by diabetes or short-term stress;
decreases may be found in cases of neurologic disease or
malnutrition

Increases in these enzymes may indicate liver damage, muscle

damage, or increased thyroid gland activity