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INTRODUCTION TO

CLINICAL HEMATOLOGY
Vet . Heba Yasser

BLOOD

Blood is the biological fluid most routinely


analyzed in the laboratory.

BLOOD

Blood is a fluid tissue that circulates through


the vascular channels carrying the necessities for
life to all cells of the body, and it receives the
waste products of metabolism for transport to the
organs of excretion.

BLOOD COMPOSITION
The cellular
portion
is assessed in
the clinical
hematology
laboratory

The liquid
portion
evaluated in the
clinical
biochemistry
laboratory.

AIM OF BLOOD EXAMINATION:


1.
2.
3.
4.
5.

Screening procedure for general


health.
Diagnosis of different diseases.
To asses the body's ability to fight
infection (cellular immunity).
Evaluation of certain disease
condition.
Follow up the response to certain
therapy.

PHYSIOLOGICAL CONSIDERATIONS IN
INTERPRETATION OF BLOOD VALUES
1. Emotional stress: facing different environment in
the veterinary clinic, Blood should be collected when
the animal is at rest.

2. Exercise: especially in animals which have a


responsive spleen, like horse.

PHYSIOLOGICAL CONSIDERATIONS
IN INTERPRETATION OF BLOOD
VALUES
3. Diet: Samples shouldn't be collected soon or

shortly after feeding. Animal is preferred to be


fasted(8-10 hours) before sampling to avoid
processing problems associated with
postprandial lipemia.
4. Drugs
5. Age, sex and breed

Emotional
stress:
Exercise:

Prolonged
exercise:
Diet: after
meals
Starvation:

Physiological function

Altered test

Hyperglycemia
Lecucytosis
Lymphocytosis in
cats
Hypoxia
Release of blood
induce spleenic cells into
contraction
circulation
increases activity of muscular
enzymes
lipemic

Blood sugar test


T.L.C.
D.L.C

response
Epinephrine

Drugs:
Glucocorticoids

PCV
TLC
CK, AST, LDH

Blood sugar test


Lipids test
Decreased BUN &
serum albumin
TLC, DLC,BUN
Liver enz. Activity ALP, ALT
alter tests to detect immune-mediated
diseases.

Physiological
Drugs:
exogenous
insulin
Antibiotics :
Cephalosporins
Age, sex and
breed

response
decreases
serum glucose

function

Altered test
Blood glucose
test and serum
potassium

increased
creatinine
concentration
Significantly influences some of the blood values.

COLLECTION AND HANDLING THE


BLOOD SPECIMEN
The selection of the proper container and the
proper anticoagulant must be made prior to
specimen collection.
1. Containers (equipment):
Syringes and Vaccutainers
Are of variable sizes.
Should be chemically clean and dry.
Glass vials fitted with rubber or polyethylene
stoppers are recommended.

2. Anticoagulants

SYRINGES : PRECAUTIONS
The

blood must be transferred quickly into


the anticoagulant to prevent initiation of
the clotting mechanism.

Using

calibrated syringe to ensure the


collection of an exact volume of blood
suitable to the amount of the
anticoagulant contained in the sample
vial.

Old metallic syringe

Plastic disposable syringe

VACUTAINERS (VACUUM TUBES)


Are plain, anticoagulated or containing clotting
accelerator.
The stoppers' color indicates the contained
anticoagulant and use.

TECHNIQUE AND PRECAUTIONS


OF BLOOD COLLECTION
I. Precautions before blood collection:
1.
2.
3.

4.

Select the site of sampling; free from cyanosis,


inflammation, or edema.
Shave the site of sampling; especially in longhaired animals.
Sterilize the site of sampling by 70 % alcohol.
(Rubbing with alcohol makes the vein more
clearly outlined).
Anesthesia and skin incision may be required; in
the rat, guinea pig, and rabbit.

II. AMOUNT OF BLOOD


REQUIRED:
Depends

on the requested test(s), and the


method used for conducting the test.
As a general role; it is usually safe to take
about 0.5 ml blood/kg body weight in all
species.
5 ml in large animals and 2 ml of blood in
small animals are sufficient for routine blood
study.

1. SMALL AMOUNT OF BLOOD:


Site:
Capillary bed of the skin,
Clipping the toe nail, or
Bricking the marginal ear vein.
Aim:
Small amount of blood is collected if only RBCs count,
hemoglobin concentration, PCV, leucocytes count, or
blood smear examinations are needed.
Technique:
Sudden sharp stab using an automatic lancet, an
ordinary lancet, or sterile needle.
Exclude the first exuding blood drop (contaminated with
the tissues) and collect the second.

Lancets

2. LARGE AMOUNT OF BLOOD


Large

blood samples are drown from major

veins.
Occlude the vein using digital pressure or
tourniquet.
Stretch the skin across the vein.
Insert the needle, while the bevel is directed
upward.
Apply gentile traction or suction of blood to
avoid collapse of the vein.
Release the digital pressure, or remove the
tourniquet.
Gently withdraw the blood by the plungers.

Jugular vein

SITES OF BLOOD COLLECTION


Horse, Sheep, Goat and Camel:
Jugular vein (middle third; more superficially
exposed).
Cow:
Jugular vein (middle third; more superficially
exposed), Tail venipuncture (coccygeal vein), or the
mammary vein.
Dog:
Cephalic (most commonly), jugular (occasionally),
saphenous, or tibial veins.
Cat:
Cephalic (most commonly), jugular (occasionally), or
femoral veins, or clipping of the toe nail.

Sampling from cephalic vein


in cats

Jugular vein in dog

Sampling from
coccogyeal artery
in cow

Jugular vein in
lamb

Jugular vein in horse

SITES OF BLOOD COLLECTION


Rabbit, Guinea pig:
Heart, ear vein, or jugular vein (occasionally).
Pig:
Anterior vena cava, ear vein, or jugular vein
(occasionally).
Rat, Mouse:
Tail amputation (the commonest), retro orbital
venous plexus, or the heart.
Birds:
Wing vein, cutaneous ulnar vein, or brachial veins,
or the heart.

Ear vein in rabbit

Wing vein in bird

Sampling in mice

HANDLING OF THE COLLECTED


BLOOD SAMPLE

Blood samples should be processed as soon as


possible after collection.

On standing; the cells settle down and separate


from the plasma,
so it is necessary to mix the sample thoroughly
each time a portion is removed for a test.

HANDLING OF THE COLLECTED


BLOOD SAMPLE
The blood film is best made immediately from
fresh blood, 15 minute.
OR as soon as possible from anticoagulated blood,
preferably within an hour of sampling, as WBCs
undergo degenerative changes due to aging of the
cells.

HANDLING OF THE COLLECTED


BLOOD SAMPLE
Blood can be kept at room temperature for 1-2
hours.
If blood examination is to be postponed for
several hours or overnight; make blood films
immediately, perform ESR, and then refrigerate
the sample. The cell remains stable at 4o C for 24
hours.
An ice box or cold packs should be used to
transport blood samples to a distant laboratory.

IDENTIFICATION OF THE COLLECTED


BLOOD
SAMPLE
Specimens
to be sent to the laboratory should be
completely identified.

A complete history should be included with each


sample.

ANTICOAGULANTS
Hematological examination requires blood in
liquid form.
Immediately after withdrawal; blood must be
thoroughly mixed with the anticoagulant to
prevent clotting.
The common used Anticoagulants are: Heparin,
EDTA, Sodium fluoride, Sodium citrate and
double oxalates.

Mode of
action

Heparine
EDTA
Sod.Citrate
Sod.flouride
Double
oxalate

Dose

Recommended
uses

Advantages

Disadvantages

CAUSES OF SPECIMEN SPOILAGE


1. Hemolysis: Breakdown of the cellular elements
of the blood.
Effect of hemolysis on lab. Results:
Hemolysis interferes with the laboratory
investigations which are measured
colorimetrically; either by:
a) changing the optical density
b)releasing intracellular components. causing false
increase or decrease in the concentration of the
analytes

CAUSES OF HEMOLYSIS
A) Before withdrawal:
Using wet syringe or needle.
Using needle of small gauge.
B) During withdrawal:
Rapid withdrawal of the blood.
Moving the needle inside the vein.
Placing the blood into the vacuum tubes too
quickly.

C) After withdrawal:
o

Dispensing the blood without removing the needle.

Incorrect dose of the anticoagulant.

Vigorous mixing of the blood with the anticoagulant.


(Rough handling of specimen).

Centrifuging blood for longer time and at higher speed.

Exposing blood to extreme heat or cold.

Storing whole blood samples in freezing temperature.

Bacterial or chemical contamination.

CAUSES OF SPECIMEN SPOILAGE


2. Lipemia: The presence of high concentration of
lipids in the blood.
Effects of lipemia on laboratory tests:
Lipemia enhances hemolysis.
Lipemia produce false-high values of Hb, and so
incorrect MCH and MCHC.
Turbidity of the serum impair end-point
spectrophotometric determinations.

LIPEMIA
Lipemia falsely decrease serum sodium and
potassium measurements by flame photometry.
Lipemia falsely increase plasma proteins
measurement by refracto- metry.
Lipemia causes false decrease in cholesterol in
cats.

CAUSES OF SPECIMEN SPOILAGE


3. Clotting: The blood loses its liquid form and
becomes clotted.
Causes of clotting:
Taking too much time in obtaining the blood; so
the blood has already started to clot before being
mixed with the anticoagulant.
Using no anticoagulant.
Failure to mix the blood with the anticoagulant.
Using inadequate dose of the anticoagulant.

CAUSES OF SPECIMEN SPOILAGE


4. Decomposition: due to overgrowth of bacteria
or mould. The blood become unfit for
examination.
Causes of decomposition:
High temperature.
Bacterial contamination.
Contamination with the soil or fecal materials.

CAUSES OF SPECIMEN SPOILAGE


5. Desiccation (Drying):
Drying of blood sample.
Causes of sample drying:
Failure to back the specimen properly.
The principal packing faults are:
a) too small sample in a too large container.
b) leaving the container opened to air.