1. Venipuncture and Clinical Pathology for Reptiles

2. Introduction ► Reptiles  stoic  evolved to mask illness ► Treat n’ run? ► Diagnostic Tests  to identify physiologic anomalies

3. Weight the reptile first

4. 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

5. Venipuncture sites: Lizards ► Ventral coccygeal/tail vein ► Ventral abdominal vein ► Jugular ► Cardiac- euthanasia ► Toe nail

6. Venipuncture sites: Snakes ► Cardiac ► Ventral tail vein

7. Venipuncture sites: Chelonians ► Jugular ► Dorsal coccygeal ► Brachial ► Subcarapacial

8. Subcarapacial

9. Venipuncture sites: Crocodilians ► Ventral tail ► Ventral abdominal ► Cardiac ► Supravertebral

10. 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*

11. 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

12. 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

13. 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

14. 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

15. Hematology ► Erythrocytes  abnormal shapes and sizes ► regeneration  basophilic stippling ► anemia, iron deficiency  hypochromatosis- ► iron deficiency anemia, chronic blood loss

16. Hematology ► Blood parasites  common wild caught specimens  evaluate overall health ► anemia? ► regenerative

17. Hematology: Leukocytes ► Circulating pool very large  reference ranges ► 5.0-15.0 x 10 3

18. Hematology: Granulocytes ► Heterophils  fusiform granules  large round, acentric nucleus  function- ► phagocytosis  heterophilia ► inflammation, stress ► seasonal  increase summer

19. Hematology: Granulocytes ► Toxic heterophils  suggestive of infectious cause  indicative of severity  changes ► degranulation ► cell swelling ► cytoplasmic vacuolization

20. Hematology : Granulocytes ► Eosinophil  large, round cells  spherical granules  nucleus centrally located  function- ► Parasitemia ► Allergy ► Inflammation  seasonal variation ► increased winter

21. Hematology : Granulocytes ► Basophils  small, spherical cells  basophilic meta- chromatic granules  function ► histamine release

22. Hematology: Lymphocyte ► Lymphocytes  small, basophilic staining  acentric nucleus  function ► B cells- Ab production ► T cells moderate immune function  seasonal variation ► decreased in winter

23. Hematology: Monocyte ► Largest WBC ► cytoplasm- blue-gray  function ► granuloma formation

24. Hematology ► Thrombocytes  central nucleus  confused with lymphocytes  functions- thrombus formation, clot, wound healing  >200,000 cells/ml

25. Injections ► Page 110 L, 159 S, 177 T ► Demonstration

26. Fluid Support ► Ideal fluid is hypotonic, non- lactated, balanced electrolyte (e.g. Normosol R) ► Maintenance fluids can be calculated at 20 ml.kg.day.

27. 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 Walkabout Farms Walkabout Farms

28. Fluid Support Pharyngostomy tubes may be needed for some chelonians Force feeding, assist feeding

29. ► Force feeding demo.

30. 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

31. Chelonians Whole-body Craniocaudal view

33. Venomous snake

34. Questions?