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Clinical Pathology Considerations for Wildlife

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Title: Clinical Pathology Considerations for Wildlife


1
Clinical Pathology Considerations for Wildlife
  • Mark A. Mitchell DVM, MS, PhD
  • University of Illinois
  • College of Veterinary Medicine

2
Introduction
  • Wildlife
  • stoic
  • evolved to mask illness
  • Treat n run?
  • Diagnostic Tests
  • to identify physiologic anomalies

3
How much blood can you collect???
4
Blood Collection
  • Blood volume
  • 8-10 of body weight of birds
  • General rule of thumb
  • 0.8-1.0 ml/100 g (avian and mammal)
  • 0.5-0.8 ml/100 g (reptiles)

Where do you collect it from????
5
Venipuncture sites
6
Venipuncture sites
7
Venipuncture sites
8
Venipuncture Sites Lizards
Ventral tail vein
9
Venipuncture sites Lizards
Jugular vein
  • Ventral coccygeal/tail vein
  • Ventral abdominal vein
  • Cardiac- euthanasia
  • Toe nail- never

10
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11
Ventral abdominal vein
12
Venipuncture sites Snakes
Venipuncture Sites Snakes
  • Cardiac
  • Ventral tail vein
  • Palatine
  • Jugular

Cardiac
13
Ventral tail vein
14
Jugular vein
15
Palatine vein
16
Venipuncture sites Chelonians
  • Jugular
  • Dorsal coccygeal
  • Brachial
  • Subcarapacial

17
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19
Blood collection tubes
  • CBC
  • EDTA or lithium heparin
  • Plasma chemistry
  • Lithium heparin

EDTA
LH
20
Hematology
  • Evaluating the packed cell volume
  • Reference ranges
  • Birds40-55
  • Mammals 30-55
  • Reptiles 20-40
  • Low values?
  • Chronic inflammation
  • Trauma blood loss
  • Immune mediated
  • Not making
  • When are you concerned?
  • Transfusion??

21
Hematology
  • Evaluating the packed cell volume
  • Buffy coat
  • anecdotal used to estimate WBC (mammals)
  • limited samples suggest not useful
  • Lawton and Divers, 1999
  • Serum
  • appearance

22
Hematology Slide Preparation
  • Techniques
  • Standard slide technique
  • cell destruction in avian and reptile patients
  • 1 drop 22 bovine albumin 5 drops blood

23
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24
Hematology Slide Preparation
25
Hematology Evaluating the erythron
  • Total Red Blood Cell Count
  • Manual techniques
  • Erythrocyte Unopette system
  • Natt and Herricks solution

26
Hematology
  • Erythrocytes
  • mature cells
  • ellipsoid with central nucleus
  • cytoplasm- orange-pink
  • immature cells
  • small, round cells increased NC ratio
  • basophilic cytoplasm

Mature
Immature
27
Hematology
  • Erythrocytes
  • abnormal shapes and sizes
  • regeneration
  • basophilic stippling
  • anemia, iron deficiency
  • hypochromatosis-
  • iron deficiency anemia, chronic blood loss

28
Hematology
  • Blood parasites
  • common wild caught specimens
  • evaluate overall health
  • evaluate erythron
  • anemia?
  • regenerative

29
Hematology Leukocytes
  • Birds and Reptiles
  • Manual counts necessary because of nucleated RBCs
    and thrombocytes
  • Natt Herricks technique
  • Stains blue
  • Small lymphocytes and thrombocytes appear similar
  • Phloxine B solution
  • Stains heterophils and eosinophils red
  • Total WBC count determined after differential

30
Hematology Leukocytes
  • Counting white blood cells
  • Hemocytometer
  • mammals recommend count all 64 squares
  • Doxley, 1989
  • avian count 4 large squares
  • Hawkey and Samour, 1988

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32
Hematology Leukocytes
  • Counting white blood cells
  • Hemocytometer
  • mammals recommend count all 64 squares
  • Doxley, 1989
  • avian count 4 large squares
  • Hawkey and Samour, 1988
  • reptiles count 9 large squares
  • Campbell, 1996
  • reptiles no difference between counting 10 or 40
    large squares
  • Lawton and Divers, 1999

33
Complete blood counts
  • Cell types
  • Heterophils-Neutrophils
  • Eosinophils
  • Basophils
  • Lymphocytes
  • Monocytes
  • Measures current status
  • Elevated?
  • Time frame?
  • Iguana study

34
Hematology Leukocytes
  • Circulating pool very large
  • Reptile study
  • Salmonella SQ
  • 12 hr 45.0-76.0 x 103
  • 24-48 h returned to normal

35
Clinical pathology
  • Complete blood count
  • Methods
  • Eosinophil unopette estimate and differential
  • Misclassification
  • Sample quality
  • Volume
  • Transport
  • Lab/Technician
  • Sensitivity and specificity variable with
    application

36
Effect of sampling site??
37
Clinical pathology
  • Normal ranges?????
  • WBC
  • Mean 4.8 x 103
  • SD 2.5
  • gt 2 SD WBC0?
  • Best measure central tendency?

38
Clinical pathology
  • Data presentation Percentages
  • White blood cell count 4.8-15.6 x 103
  • Heterophils 48-75
  • 2.3 x 103 (48 of 4.8)
  • 11.7 x 103 (75 of 15.6)
  • Prefer to use absolutes

39
Mitchells Rules
  • WBC 5-15,000 cells/ml
  • Heterophil Lymphocyte ratio
  • 11-31
  • Raptors- heterophil predominant cell
  • Lymphocytes
  • lt1,500 cells/ml absolute lymphopenia
  • Monocytes
  • lt 500 cells/ml normal
  • 500-1,000 cells/ml stress
  • gt1,500 cells/ml chronic inflammation
  • Eosinophils and basophils
  • lt 500 cells/ml

40
Hematology Granulocytes
  • Heterophils
  • fusiform granules
  • large round, acentric nucleus
  • function-
  • phagocytosis
  • heterophilia
  • inflammation, stress
  • seasonal
  • increase summer

41
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42
Hematology Granulocytes
  • Toxic heterophils
  • suggestive of infectious cause
  • indicative of severity
  • changes
  • degranulation
  • cell swelling
  • cytoplasmic vacuolization

43
Hematology Granulocytes
  • Eosinophil
  • large, round cells
  • spherical granules
  • nucleus centrally located
  • function-
  • parasitemia
  • seasonal variation
  • increased winter

44
Hematology Granulocytes
45
Hematology Granulocytes
  • Basophils
  • small, spherical cells
  • basophilic meta-chromatic granules
  • function
  • histamine release

46
Hematology Lymphocyte
  • Lymphocytes
  • small, basophilic staining
  • acentric nucleus
  • function
  • B cells- Ab production
  • T cells moderate immune function
  • seasonal variation
  • decreased in winter

47
Hematology Monocyte
  • Largest WBC
  • cytoplasm- blue-gray
  • function
  • granuloma formation
  • antigenic processing
  • chronic inflammation

48
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49
Hematology
  • Thrombocytes
  • central nucleus
  • confused with lymphocytes
  • functions- thrombus formation, clot, wound
    healing
  • gt200,000 cells/ml

50
Plasma Chemistries
  • Reference material limited
  • Affected by seasonal, environmental, gender and
    nutritional changes
  • Sample size considerations

51
Equipment
  • Abaxis VetScan
  • Small sample volumes required
  • Whole blood or plasma
  • Portable
  • Rapid
  • No calibration time

52
Plasma Chemistries
  • Glucose
  • reference range highly variable
  • avian 100-400 mg/dl
  • hypoglycemia
  • starvation, septicemia, hepatopathies, high
    protein diets,
  • hyperglycemia
  • rare

53
Plasma Chemistries
  • Protein
  • avian 2.5-6.0 mg/dl
  • Mammal and reptile 4-8 mg/dl
  • Hyperproteinemia
  • dehydration
  • elevated globulins- serum electrophoresis
  • alpha 1, alpha 2, beta 1, beta 2, gamma
  • Hypoproteinemia
  • malnutrition, malabsorption, blood loss, hepatic
    or renal failure

54
Plasma Chemistries
  • Uric Acid
  • primary catabolic end-product of protein
  • reference range lt 10-15 mg/dl
  • hyperuricemia
  • renal disease, dehydration, gout
  • affected by diet carnivores
  • not specific to renal function

55
Plasma Chemistries
  • Sodium (140-160 mg/dl)
  • hypernatremia- dehydration, renal disease,
    re-feeding syndrome
  • hyponatremia- diarrhea, diet
  • Chloride (90-120 mg/dl)
  • hyperchloremia- dehydration, renal disease
  • Potassium (2-6 mg/dl)
  • hypokalemia diet, diarrhea
  • re-feeding syndrome- shift extra- to intracellular

56
Plasma Chemistries
  • Aspartate aminotransferase (AST)
  • present in all body tissues
  • elevated levels in liver and muscle
  • Alanine aminotransferase (ALT)
  • Hepatocytes- mammals
  • Unreliable- birds and reptiles
  • Creatine kinase (CK)
  • skeletal muscle
  • rule-out liver disease with AST

57
Plasma Chemistries
  • Calcium
  • measures total calcium
  • ionized biologically active
  • unionized bound to protein or complexed
  • reduced pH increases ionic form
  • e.g., apnea- anesthesia
  • increased likelihood for calcification

58
Plasma Chemistries
  • Calcium
  • reference ranges 8-12 mg/dl
  • calcium to phosphorus ratio
  • calcium-phosphorus product

59
Plasma Chemistries
  • Calcium
  • hypocalcemia
  • renal compromise
  • nutritional imbalance
  • certain speciesAfrican Grey
  • hypercalcemia
  • primary hyperparathyroidism
  • folliculogenesis

60
Plasma Chemistries
  • Phosphorus
  • reference range 2-6 mg/dl
  • hyperphosphatemia
  • renal disease, hypervitaminosis D, excessive
    dietary phosphorus, folliculogenesis
  • hypophosphatemia
  • starvation (re-feeding syndrome), dietary
    imbalance

61
Plasma Chemistries
  • Bile Acids
  • sensitive indicator of liver function in birds
  • elevations in bile acid- liver biopsy
  • Pre-prandial and post-prandial levels identified
    in some species of psittacines

62
Questions?
Questions??
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