C M AC M A Cooperative Medical Assistance (CMA) BLOOD SMEAR PREPARATION AND EVALUATION

C M AC M A Overview Blood Smear Preparation Blood Smear Evaluation

C M AC M A Blood Smear Preparation Materials and Supplies Sample (EDTA) Slides Fixative, Buffer and Stain Diff-Quik Water

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

C M AC M A A spreader slide is used to feather the drop of blood.

C M AC M A 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

C M AC M A Staining

C M AC M A Microscopic Evaluation 10X objective

C M AC M A Erythrocyte (RBC) RBC estimate Cell morphology

C M AC M A 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

C M AC M A Thrombocyte Platelets Small cell fragments that are responsible for blood clotting

C M AC M A Total Platelets Species Range Horse 100,000 600,000 Cattle 100,000 800,000 Sheep 250,000 750,000 Goat 300,000 600,000

C M AC M A RBC Morphology Morphology of Erythrocytes can be categorized according to: Cell arrangements Size Color Shape Presence of structures in or on the RBC

C M AC M A MCH and MCHC Normochromic describe the erythrocyte as normal range Hypochromic below the range

C M AC M A Anisocytosis -Variation in size of RBCs

C M AC M A Acanthocytes Abnormal blunt projections on the cell wall Burr or spur cells Alteration in the cell membrane

C M AC M A Shistocyte Fragmented RBCs

C M AC M A Rouleaux Formation where RBCs stack Commonly seen in horses, rarely seen in ruminants May increase with inflammatory disease

C M AC M A Poikilocytosis Abnormal shape of RBCs Variation in shape of the RBCs May represent formational defects or mechanical or toxic damage

C M AC M A Crenation Mostly an artifact of preparation and not due to cellular changes Slow drying of blood film Cell shrinkage

C M AC M A 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

C M AC M A Leukocyte (WBC) WBC estimate WBC differential WBC morphology

C M AC M A 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

C M AC M A 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 (%)

C M AC M A 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

C M AC M A WBCs Granulocytes Eosinophils Neutrophils Basophils Agranulocytes Lymphocytes Monocytes

C M AC M A Eosinophils Help in controlling allergic and or anaphylactic reactions Distinct red staining granules Segmented nucleus

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

C M AC M A Basophils Segmented nucleus Purple to blue-black granules

C M AC M A Lymphocytes Small to medium size mononuclear cells Slight cytoplasm, stains blue

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

C M AC M A Nucleated RBCs Inclusion Bodies Parasites Bacterias

C M AC M A Nucleated RBC Represent early release of immature cells during anemia.

C M AC M A Babesia Large teardrop-shaped intracellular protozoan Frequently seen in pairs

C M AC M A Rickettsial Parasite found in RBCs

C M AC M A Trypanosomes

C M AC M A Anthrax

C M AC M A ANAPLASMOSIS Rickettsial disease of ruminants