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Cooperative Medical Assistance
(CMA)
BLOOD SMEAR
PREPARATION AND EVALUATION

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Overview
• Blood Smear Preparation
• Blood Smear Evaluation

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Blood Smear Preparation
• Materials and Supplies
– Sample (EDTA)
– Slides
– Fixative, Buffer and Stain
• Diff-Quik
• Water

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A drop of blood is placed on a clean
microscope slide.
Push-smear Technique

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A spreader slide is used to “feather” the drop
of blood.

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Diff-Quik Technique
FIXATIVE About 5 minutes
Solution 1 8-10 dips
(1 second each)
Solution 2 6-8 dips
(1 second each)
Water Rinse till clean

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Staining

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Microscopic Evaluation
10X objective

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Erythrocyte (RBC)
• RBC estimate
• Cell morphology

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Total RBCs
Species Average (x106) Range (x106)
Horse 8.0 6 - 12
Cattle 7.0 5 – 10
Sheep 12.0 8 – 15
Goat 13.0 8 – 17

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Thrombocyte
• Platelets
• Small cell fragments that are responsible
for blood clotting

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Total Platelets
Species Range
Horse 100,000 – 600,000
Cattle 100,000 – 800,000
Sheep 250,000 – 750,000
Goat 300,000 – 600,000

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RBC Morphology
• Morphology of Erythrocytes can be
categorized according to:
– Cell arrangements
– Size
– Color
– Shape
– Presence of structures in or on the RBC

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MCH and MCHC
• Normochromic
– describe the
erythrocyte as
normal range
• Hypochromic
– below the range

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Anisocytosis
-Variation in size of RBC’s

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Acanthocytes
• Abnormal blunt projections on the cell wall
• Burr or spur cells
• Alteration in the cell membrane

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Shistocyte
• Fragmented RBC’s

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Rouleaux
• Formation where
RBCs stack
• Commonly seen in
horses, rarely seen in
ruminants
• May increase with
inflammatory disease

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Poikilocytosis
• Abnormal shape of
RBC’s
• Variation in shape of
the RBCs
• May represent
formational defects or
mechanical or toxic
damage

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Crenation
• Mostly an artifact of
preparation and not
due to cellular
changes
• Slow drying of blood
film
• Cell shrinkage

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Target Cells
• Appearance of a
“target” or thicker,
dark-staining center
surrounded by a
light stained area
and dark periphery
• Most common with
chronic disease

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Leukocyte (WBC)
• WBC estimate
• WBC differential
• WBC morphology

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Total WBC
Species Average Range (cells/cu mm)
Horse 9,250 6,000 – 14,000
Cattle 8,000 4,000 – 12,000
Sheep 9,000 4,000 – 12,000
Goat 12,000 6,000 – 13,000

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WBC Differential
Band Segmented
Neutrophils Neutrophils Lymphocyte
Horse 0 – 1 30 – 75 25 – 60
Cow 0 – 2 15 – 45 45 – 75
Sheep Rare 10 – 50 40 – 75
Goat Rare 30 – 48 50 – 70
Differential range (%)

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WBC Differential
Average differential range (%)
Monocyte Eosinophil Basophil
Horse 1 – 8 5 – 10 0 – 3
Cow 2 – 7 2 – 20 0 – 2
Sheep 0 – 6 0 – 10 0 – 3
Goat 0 – 4 1 – 8 0 – 1

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WBCs
• Granulocytes
– Eosinophils
– Neutrophils
– Basophils
• Agranulocytes
– Lymphocytes
– Monocytes

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Eosinophils
• Help in controlling
allergic and or
anaphylactic
reactions
• Distinct red staining
granules
• Segmented nucleus

28. C M AC M A Neutrophils
• Predominant
WBC
• Phagocytic and
bactericidal
• 10 hour life span
in circulating
blood

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Basophils
• Segmented nucleus
• Purple to blue-black granules

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Lymphocytes
• Small to medium size mononuclear cells
• Slight cytoplasm, stains blue

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Monocytes
• Derived from bone marrow
• Largest circulating cell
• Found only briefly in circulation before entering
tissues and becoming a macrophage

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• Nucleated RBCs
• Inclusion Bodies
• Parasites
• Bacterias

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Nucleated RBC
• Represent early release of immature cells
during anemia.

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Babesia
– Large teardrop-shaped intracellular
protozoan
– Frequently seen in pairs

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Rickettsial
Parasite found in RBCs

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Trypanosomes

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Anthrax

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ANAPLASMOSIS
Rickettsial disease of ruminants