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Venipuncture and Clinical Pathology for Reptiles. Introduction. Reptiles stoic evolved to mask illness Treat n’ run? Diagnostic Tests to identify physiologic anomalies. Weight the reptile first. Blood Collection. Blood volume: 5-8% of body weight reptiles Tolerate 1% loss
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Introduction • Reptiles • stoic • evolved to mask illness • Treat n’ run? • Diagnostic Tests • to identify physiologic anomalies
Blood Collection • Blood volume: • 5-8% of body weight reptiles • Tolerate 1% loss • General rule of thumb: • 0.5-0.8 ml/100 g (reptiles) • To heparinize or not to heparinize? • Consider time to collect • LIMIT VOLUME OF HEPARIN • May affect cell staining
Venipuncture sites: Lizards • Ventral coccygeal/tail vein • Ventral abdominal vein • Jugular • Cardiac- euthanasia • Toe nail
Venipuncture sites: Snakes • Cardiac • Ventral tail vein
Venipuncture sites: Chelonians • Jugular • Dorsal coccygeal • Brachial • Subcarapacial
Venipuncture sites: Crocodilians • Ventral tail • Ventral abdominal • Cardiac • Supravertebral
Blood collection tubes: Ethylene diamine tetra acetic acid • CBC and plasma chemistry • use microtainers • less likely to dilute sample • do not overfill- microclots • disadvantages: • hemolyzes RBC’s: especially tortoises • elevated potassium* • decreased calcium*
Blood collection tubes: Lithium heparin • CBC or plasma chemistries • less damaging to cells • centrifuge immediately • cells will consume glucose • potassium and phosphorus increase from leakage
Blood collection tubes: No anticoagulant • Red top tube • centrifuge to collect serum • serum may form gelatinous clot • may lose sample • continue to centrifuge, may help • more common in glass tubes • microtainer: plastic
Hematology • Evaluating the packed cell volume • Reference ranges: • 20-40% (reptile) • 40-55% (avian) • 30-55% (mammalian; ferret 45-58%) • Buffy coat: • anecdotal used to estimate WBC (mammals) • limited samples suggest not useful • Lawton and Divers, 1999 • Serum • appearance
Hematology: • Total Red Blood Cell Count • Manual techniques: • Erythrocyte Unopette system • Erythrocytes • mature cells • ellipsoid with central nucleus • cytoplasm- orange-pink • immature cells • small, round cells • basophilic cytoplasm
Hematology • Erythrocytes • abnormal shapes and sizes • regeneration • basophilic stippling • anemia, iron deficiency • hypochromatosis- • iron deficiency anemia, chronic blood loss
Hematology • Blood parasites • common wild caught specimens • evaluate overall health • anemia? • regenerative
Hematology: Leukocytes • Circulating pool very large • reference ranges • 5.0-15.0 x 103
Hematology: Granulocytes • Heterophils • fusiform granules • large round, acentric nucleus • function- • phagocytosis • heterophilia • inflammation, stress • seasonal • increase summer
Hematology: Granulocytes • Toxic heterophils • suggestive of infectious cause • indicative of severity • changes • degranulation • cell swelling • cytoplasmic vacuolization
Hematology : Granulocytes • Eosinophil • large, round cells • spherical granules • nucleus centrally located • function- • Parasitemia • Allergy • Inflammation • seasonal variation • increased winter
Hematology : Granulocytes • Basophils • small, spherical cells • basophilic meta-chromatic granules • function • histamine release
Hematology: Lymphocyte • Lymphocytes • small, basophilic staining • acentric nucleus • function • B cells- Ab production • T cells moderate immune function • seasonal variation • decreased in winter
Hematology: Monocyte • Largest WBC • cytoplasm- blue-gray • function • granuloma formation
Hematology • Thrombocytes • central nucleus • confused with lymphocytes • functions- thrombus formation, clot, wound healing • >200,000 cells/ml
Injections • Page 110 L, 159 S, 177 T • Demonstration
Fluid Support • Ideal fluid is hypotonic, non-lactated, balanced electrolyte (e.g. Normosol R) • Maintenance fluids can be calculated at 20 ml.kg.day.
Fluid Support • SQ, ICe, IV, IO, PO are all effective, choice of route depends on severity of illness and logistics of individual animals • Always inject between scales • Liquid enteral nutrition (e.g. Ensure, Sustacal, useful for recovery from long-term anorexia, may be mixed with vegetables or fruits for herbivores) • Critical Care Herbivore Diet by Oxbow Hay Co. or enterals from Walkabout Farms
Fluid Support Pharyngostomy tubes may be needed for some chelonians Force feeding, assist feeding
Radiology • Lateral, DV, AP • Horizontal beam if possible • Consider mammography or dental film • GI contrast • IVP- recently described, usefulness needs to be determined. • Page 110 L, 150 S, 175 T