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Venipuncture and Venipuncture and Clinical Pathology for Clinical Pathology for
ReptilesReptiles
IntroductionIntroduction
► ReptilesReptiles stoicstoic evolved to mask evolved to mask
illnessillness
► Treat n’ run?Treat n’ run?►Diagnostic TestsDiagnostic Tests
to identify to identify physiologic physiologic anomaliesanomalies
Weight the reptile first Weight the reptile first
Blood CollectionBlood Collection
►Blood volume:Blood volume: 5-8% of body weight reptiles5-8% of body weight reptiles
►Tolerate 1% lossTolerate 1% loss
►General rule of thumb: General rule of thumb: 0.5-0.8 ml/100 g (reptiles)0.5-0.8 ml/100 g (reptiles)
►To heparinize or not to heparinize?To heparinize or not to heparinize? Consider time to collectConsider time to collect LIMIT VOLUME OF HEPARINLIMIT VOLUME OF HEPARIN May affect cell stainingMay affect cell staining
Venipuncture sites: LizardsVenipuncture sites: Lizards
► Ventral coccygeal/tail Ventral coccygeal/tail veinvein
► Ventral abdominal veinVentral abdominal vein
► JugularJugular
► Cardiac- euthanasiaCardiac- euthanasia
► Toe nailToe nail
Venipuncture sites: SnakesVenipuncture sites: Snakes
► CardiacCardiac
► Ventral tail veinVentral tail vein
Venipuncture sites: Venipuncture sites: CheloniansChelonians
► JugularJugular
►Dorsal coccygealDorsal coccygeal
► BrachialBrachial
► SubcarapacialSubcarapacial
SubcarapacialSubcarapacial
Venipuncture sites: Venipuncture sites: CrocodiliansCrocodilians
► Ventral tailVentral tail
► Ventral abdominalVentral abdominal
► CardiacCardiac
► SupravertebralSupravertebral
Blood collection tubes: Blood collection tubes:
Ethylene diamine tetra acetic acidEthylene diamine tetra acetic acid ►CBC and plasma CBC and plasma
chemistrychemistry►use microtainersuse microtainers
less likely to dilute less likely to dilute samplesample
do not overfill- do not overfill- microclotsmicroclots
►disadvantages:disadvantages: hemolyzes RBC’s: hemolyzes RBC’s:
especially tortoisesespecially tortoises elevated elevated
potassium*potassium* decreased calcium*decreased calcium*
Blood collection tubes: Blood collection tubes: Lithium heparinLithium heparin
► CBC or plasma CBC or plasma chemistrieschemistries less damaging to less damaging to
cellscells centrifuge centrifuge
immediately immediately ►cells will consume cells will consume
glucoseglucose►potassium and potassium and
phosphorus increase phosphorus increase from leakagefrom leakage
Blood collection tubes: Blood collection tubes: No anticoagulantNo anticoagulant
►Red top tubeRed top tube centrifuge to collect centrifuge to collect
serumserum serum may form serum may form
gelatinous clotgelatinous clot►may lose samplemay lose sample►continue to centrifuge, continue to centrifuge,
may helpmay help►more common in glass more common in glass
tubestubes microtainer: plasticmicrotainer: plastic
HematologyHematology
►Evaluating the packed cell volumeEvaluating the packed cell volume►Reference ranges: Reference ranges:
20-40% (reptile)20-40% (reptile) 40-55% (avian)40-55% (avian) 30-55% (mammalian; ferret 45-58%)30-55% (mammalian; ferret 45-58%)
►Buffy coat:Buffy coat: anecdotal used to estimate WBC (mammals)anecdotal used to estimate WBC (mammals) limited samples suggest not usefullimited samples suggest not useful
►Lawton and Divers, 1999Lawton and Divers, 1999
►SerumSerum►appearanceappearance
Hematology:Hematology:
► Total Red Blood Cell CountTotal Red Blood Cell Count Manual techniques: Manual techniques:
►Erythrocyte Unopette Erythrocyte Unopette systemsystem
► ErythrocytesErythrocytes mature cellsmature cells
►ellipsoid with central ellipsoid with central nucleusnucleus
►cytoplasm- orange-cytoplasm- orange-pinkpink
immature cells immature cells ►small, round cellssmall, round cells►basophilic cytoplasmbasophilic cytoplasm
HematologyHematology
►ErythrocytesErythrocytes abnormal shapes and sizesabnormal shapes and sizes
►regenerationregeneration
basophilic stipplingbasophilic stippling►anemia, iron deficiencyanemia, iron deficiency
hypochromatosis-hypochromatosis-►iron deficiency anemia, chronic blood lossiron deficiency anemia, chronic blood loss
HematologyHematology
► Blood parasitesBlood parasites common wild caught common wild caught
specimensspecimens evaluate overall evaluate overall
healthhealth►anemia?anemia?► regenerativeregenerative
Hematology: LeukocytesHematology: Leukocytes
► Circulating pool Circulating pool very largevery large reference rangesreference ranges
►5.0-15.0 x 105.0-15.0 x 1033
Hematology: GranulocytesHematology: Granulocytes
►HeterophilsHeterophils fusiform granulesfusiform granules large round, acentric large round, acentric
nucleus nucleus function-function-
►phagocytosisphagocytosis
heterophiliaheterophilia► inflammation, stress inflammation, stress ►seasonalseasonal
increase summerincrease summer
Hematology: GranulocytesHematology: Granulocytes
► Toxic heterophilsToxic heterophils suggestive of suggestive of
infectious causeinfectious cause indicative of severityindicative of severity changeschanges
►degranulationdegranulation►cell swellingcell swelling►cytoplasmic cytoplasmic
vacuolizationvacuolization
Hematology : GranulocytesHematology : Granulocytes
► EosinophilEosinophil large, round cellslarge, round cells spherical granulesspherical granules nucleus centrally locatednucleus centrally located function-function-
► ParasitemiaParasitemia► AllergyAllergy► InflammationInflammation
seasonal variationseasonal variation► increased winterincreased winter
Hematology : GranulocytesHematology : Granulocytes
► BasophilsBasophils small, spherical cellssmall, spherical cells basophilic meta-basophilic meta-
chromatic granuleschromatic granules functionfunction
►histamine releasehistamine release
Hematology: LymphocyteHematology: Lymphocyte
► LymphocytesLymphocytes small, basophilic small, basophilic
stainingstaining acentric nucleusacentric nucleus functionfunction
►B cells- Ab productionB cells- Ab production►T cells moderate T cells moderate
immune functionimmune function
seasonal variationseasonal variation►decreased in winterdecreased in winter
Hematology: MonocyteHematology: Monocyte
► Largest WBCLargest WBC►cytoplasm- blue-gray cytoplasm- blue-gray
functionfunction►granuloma formationgranuloma formation
HematologyHematology
► ThrombocytesThrombocytes central nucleuscentral nucleus confused with confused with
lymphocyteslymphocytes functions- thrombus functions- thrombus
formation, clot, formation, clot, wound healingwound healing
>200,000 cells/ml>200,000 cells/ml
Injections Injections
►Page 110 L, 159 S, 177 TPage 110 L, 159 S, 177 T►Demonstration Demonstration
Fluid Support Fluid Support
► Ideal fluid is Ideal fluid is hypotonic, non-hypotonic, non-lactated, lactated, balanced balanced electrolyte (e.g. electrolyte (e.g. Normosol R) Normosol R)
►Maintenance fluids Maintenance fluids can be calculated at can be calculated at 20 ml.kg.day. 20 ml.kg.day.
Fluid SupportFluid Support► SQ, ICe, IV, IO, PO are all SQ, ICe, IV, IO, PO are all
effective, choice of route effective, choice of route depends on severity of depends on severity of illness and logistics of illness and logistics of individual animals individual animals
► Always inject between scales Always inject between scales ► Liquid enteral nutrition (e.g. Liquid enteral nutrition (e.g.
Ensure, Sustacal, useful for Ensure, Sustacal, useful for recovery from long-term recovery from long-term anorexia, may be mixed with anorexia, may be mixed with vegetables or fruits for vegetables or fruits for herbivores) herbivores)
► Critical Care Herbivore Diet Critical Care Herbivore Diet by Oxbow Hay Co. or by Oxbow Hay Co. or enterals from enterals from Walkabout Farms
Fluid SupportFluid Support
Pharyngostomy tubes may be needed for Pharyngostomy tubes may be needed for some chelonians some chelonians Force feeding, assist feedingForce feeding, assist feeding
►Force feeding demo. Force feeding demo.
Radiology Radiology
► Lateral, DV, AP Lateral, DV, AP ► Horizontal beam if Horizontal beam if
possiblepossible ► Consider mammography Consider mammography
or dental film or dental film ► GI contrast GI contrast ► IVP- recently described, IVP- recently described,
usefulness needs to be usefulness needs to be determined. determined.
► Page 110 L, 150 S, 175 TPage 110 L, 150 S, 175 T
CheloniansCheloniansWhole-body Craniocaudal Whole-body Craniocaudal
viewview
Venomous snake Venomous snake
Questions? Questions?