1	AVIAN HEMATOLOGY Sarah E. Snead, LVT The Wildlife Center of Virginia
2	Sampling Maximum safe volume is 1% of body weight (10% of blood volume) Use TB syringe and 25g x 5/8” needle for most species (22g x 1” for large birds) Coat syringe with Heparin Sodium (1000u/ml)
3	Complete Blood Count (CBC) Packed cell volume (PCV) from hematocrit Total plasma protein (TP) Total leukocyte count (WBC) Differential Glucose
4	Venipuncture Sites Jugular vein Best site in small birds Right usually larger than left Basilic vein Best site in larger raptors Hematoma common Medial metatarsal vein Best site in waterfowl, vultures Hematoma uncommon
5	Processing Sample Make smear immediately Always fill 2 hematocrit tubes Total leukocyte count uses eosinophil Unopette method Glucose may be performed on hand held glucometer (Accu-Chek Compact)
6	Blood Smear Standard wedge technique can be used Tends to cause marked smudging of cells Preferred method is slide and long coverglass Minimizes cellular damage Drop of blood on slide, drop coverglass, pull apart horizontally, air dry, stain 2 coverglass technique possible Technique same as above Beware of breakage Stain with 3-step quick stain (Diff-Quik)
7	Hematocrit Easy, practical, very useful 35-55% normal for most species Total plasma protein obtained by breaking tube and placing drop of plasma on refractometer 3.5-5.5 g/dl normal for most species
8	Total Leukocyte Count An indirect method Uses eosinophil Unopette and hemacytometer with improved Neubauer ruling
9	Total Granulocyte Count Fill Unopette with blood and mix well Load both sides of hemocytometer ASAP Let stand 5 to 10 minutes Count all dark pink, refractile cells in all 9 primary squares on both sides of hemocytometer Multiply total cells counted x 1.1 x 16 = total granulocyte count Total leukocyte count calculated after differential
10	Total Leukocyte Count Use granulocyte count and differential to calculate % mononuclear cells ÷ % granulocytes x total # granulocytes (#cells counted on hemocytometer x 1.1 x 16) + total #granulocutes = total # leucocytes
11	Erythrocytes Oval cell with central oval nucleus Nuclear chromatin is uniformly clumped Cytoplasm stains orange-pink Nucleus stains dark purple Polychromasia denotes immature RBC 1-5% normal for avians > 5% may indicate regenerative anemia May see rubricyte or earlier Mitotic figures possible
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16	Leukocytes Granulocytes Heterophil- avian equivalent of neutrophil Eosinophil Basophil Mononuclear cells Lymphocyte- may be small, medium, large Monocyte Thrombocytes
17	Heterophil Most common cell in many species Round cell, colorless cytoplasm, rod-shaped brick red granules Mature cells have lobed nucleus Nucleus may be partially hidden by cytoplasmic granules Granules may be round in some species
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24	Heterophil Abnormalities Degranulation may be seen May indicate toxicity May be staining artifact Band cells common in ill birds Toxic changes may be seen Degranulation Vacuolization Dark granules present
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30	Eosinophil Round cell, pale blue cytoplasm, round red granules Lobed nucleus Nucleus often more clear and bluer than heterophil Granules vary in size and coloration between species Granules may be rod-shaped in some species
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36	Basophil Often seen in some avian species Round cell, eccentric or central nucleus Round dark blue/black granules with Wright’s and other fine stains Granules frequently do not stain with quick stains (Diff-Quik)
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40	Lymphocyte Most common cell in some species Round cell , round central nucleus May be irregularly shaped from molding to adjacent cells Nuclear chromatin densely clumped Amount of cytoplasm varies according to size Usually a high nuclear : cytoplasm ratio Cytoplasm pale blue, homogeneous
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45	Lymphocyte Abnormalities Reactive lymphocyte Deeply basophilic cytoplasm Lymphocyte with azurophilic granules High numbers may be abnormal Lymphocytes with scalloped borders High numbers may be abnormal Plasma cell Large lymphocytes with eccentric nuclei and prominent Golgi apparatus (clear area next to nucleus) Immature lymphocytes
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50	Monocyte Large, sometimes irregularly shaped cell Nucleus may be round to dented (kidney bean shape) Nuclear chromatin finely granular Cytoplasm blue gray, dust like granulation Vacuoles may be present More cytoplasm than nucleus (low nuclear : cytoplasm ratio)
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54	Thrombocyte Avian equivalent of platelet Small oval cell Dense, darkly staining nuclear material Clear cytoplasm may appear reticulated or contain reddish granules or vacuoles Tend to clump
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59	Hemoparasites Common in wild birds Plasmodium- avian malaria, can be pathogenic, parasite of erythrocytes Hemoproteus- low pathogenicity in most species, parasite of erythrocytes Leukocytozoan- may be pathogenic in waterfowl, turkeys, and young raptors, distorts host cell, thought to be parasite of erythrocytes Atoxoplasma- most often found in passerines, parasite of mononuclear cells
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66	QUIZ
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81	References Campbell, Terry W. , Avian Hematology and Cytology, Iowa State University Press, 1988. Dein, F. Joshua, Laboratory Manual of Avian Hematology, Association of Avian Veterinarians, 1984. Fudge, Alan M., Laboratory Medicine Avian and Exotic Pets, W. B. Saunders Company, 2000. Hawkey, C. M. and T. B. Dennett, Comparative Veterinary Hematology, Iowa State University Press, 1989. Jones, Michael P., Avian Clinical Pathology, in The Veterinary Clinics of North America, Exotic Animal Practice, Volume 2, Number 3, W. B. Saunders Company, September 1999.