Diagnostic sampling and laboratory tests

Nicole Stacy; Darryl Heard; Jim Wellehan

doi:10.22233/9781905319794.8

British Small Animal Veterinary Association , 115 (2019); https://doi.org/10.22233/9781905319794.8

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Diagnostic sampling and laboratory tests

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Authors: Nicole Stacy, Darryl Heard and Jim Wellehan
From: BSAVA Manual of Reptiles
Item: Chapter 8, pp 115 - 133
DOI: 10.22233/9781905319794.8
Copyright: © 2019 British Small Animal Veterinary Association
Publication Date: March 2019

Abstract

Accurate collection, handling, processing and submission of appropriate diagnostic specimens are powerful diagnostic tools and essential components of reptile medicine. This chapter describes techniques for taking blood, tissue, faecal and urine samples, and for performing laboratory tests in reptiles. Sampling for toxicological analysis and infectious agents is also covered.

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Figures

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8.1

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8.1 (a) Location of the jugular vein in a Chinese box turtle. (b) Venepuncture of the jugular vein in an Eastern box turtle. (Courtesy of Paul Raiti)

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8.2

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8.2 Venepuncture of the occipital sinus in an Aldabran tortoise. (Courtesy of Paul Raiti)

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8.3

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8.3 Venepuncture of the supravertebral (subcarapacial) vein of a leopard tortoise for propofol administration. (Courtesy of Paul Raiti)

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8.4

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8.4 Venepuncture of the brachial vein in an African spurred tortoise. (Courtesy of Paul Raiti)

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8.5

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8.5 Venepuncture of the dorsal coccygeal vein in a common snapping turtle. (Courtesy of Paul Raiti)

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8.6

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8.6 Cardiocentesis in a tortoise. The head is to the left. (Courtesy of Paul Raiti)

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8.7

Venepuncture of the ventral coccygeal vein in a green iguana: (a) ventral approach; (b) lateral approach. The syringe contains propofol. (Courtesy of Paul Raiti) © 2019 British Small Animal Veterinary Association

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8.7 Venepuncture of the ventral coccygeal vein in a green iguana: (a) ventral approach; (b) lateral approach. The syringe contains propofol. (Courtesy of Paul Raiti)

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8.8

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8.8 Venepuncture of the brachial plexus in a green iguana. (Courtesy of Paul Raiti)

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8.9

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8.9 Use of a blood-flow Doppler probe to locate the heart in a leaf-tailed gecko. (Courtesy of Paul Raiti)

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8.10

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8.10 Venepuncture of the ventral coccygeal vein in a Honduran milk snake. (Courtesy of Paul Raiti)

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8.11

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8.11 Palpation of the heart in a tranquilized Burmese python. The head is to the left. (Courtesy of Paul Raiti)

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8.12

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8.12 Cardiocentesis in a Burmese python. (Courtesy of Paul Raiti)

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8.13

Mature erythrocytes (some with small distinct basophilic inclusions or clear punctate vacuoles) in a blood film from a ball python. H = heterophil; L = lymphocyte; T = thrombocyte. (Wright–Giemsa stain; original magnification X100) © 2019 British Small Animal Veterinary Association

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8.13 Mature erythrocytes (some with small distinct basophilic inclusions or clear punctate vacuoles) in a blood film from a ball python. H = heterophil; L = lymphocyte; T = thrombocyte. (Wright–Giemsa stain; original magnification X100)

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8.14

Blood film from a blood python: mature erythrocytes. A = azurophil; B = basophil; H = heterophil; L = lymphocyte; P = polychromatophil; T = thrombocyte. (Wright–Giemsa stain; original magnification X100) © 2019 British Small Animal Veterinary Association

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8.14 Blood film from a blood python: mature erythrocytes. A = azurophil; B = basophil; H = heterophil; L = lymphocyte; P = polychromatophil; T = thrombocyte. (Wright–Giemsa stain; original magnification X100)

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8.15

(a) Blood film from an anaemic green turtle (PCV 12%) with synchronous erythroid regeneration. H = heterophil; L = lymphocyte; M = monocyte; P = polychromatophil; R = rebricyte (increased polychromasia, basophilic stippling). (Wright–Giemsa stain; original magnification X200). (b) Blood film from a Kemp’s ridley sea turtle with anaemia (PCV 17%) and erythroid regeneration (polychromasia, basophilic stippling and mitotic figure). (Wright–Giemsa stain; original magnification X500) © 2019 British Small Animal Veterinary Association

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8.15 (a) Blood film from an anaemic green turtle (PCV 12%) with synchronous erythroid regeneration. H = heterophil; L = lymphocyte; M = monocyte; P = polychromatophil; R = rebricyte (increased polychromasia, basophilic stippling). (Wright–Giemsa stain; original magnification X200). (b) Blood film from a Kemp’s ridley sea turtle with anaemia (PCV 17%) and erythroid regeneration (polychromasia, basophilic stippling and mitotic figure). (Wright–Giemsa stain; original magnification X500)

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8.16

(a) Blood film from a peninsula ribbon snake with squamate erythrocytic virus (SEV) infection (crystalline inclusions and purple granular aggregates within erythrocytes). (Wright–Giemsa stain; original magnification X500). (b) Blood film from an indigo snake with intraerythrocytic haemogregarines (Hepatozoon sp.). (Wright–Giemsa stain; original magnification X200). (c) A filarial worm in the blood of a monitor lizard. Possible genera include Foleyella, Piratuboides, Oswaldo, McDonadus. (Wright–Giemsa stain; original magnification X1000) © 2019 British Small Animal Veterinary Association

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8.16 (a) Blood film from a peninsula ribbon snake with squamate erythrocytic virus (SEV) infection (crystalline inclusions and purple granular aggregates within erythrocytes). (Wright–Giemsa stain; original magnification X500). (b) Blood film from an indigo snake with intraerythrocytic haemogregarines (Hepatozoon sp.). (Wright–Giemsa stain; original magnification X200). (c) A filarial worm in the blood of a monitor lizard. Possible genera include Foleyella, Piratuboides, Oswaldo, McDonadus. (Wright–Giemsa stain; original magnification X1000)

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8.17

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8.17 Blood film from a Kemp’s ridley sea turtle with thrombocyte clump (T) and lymphocytes (L). (Wright–Giemsa stain; original magnification X200)

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8.18

(a) Blood film from a green iguana with moderately toxic heterophils. (Wright–Giemsa stain; original magnification X200). (b) Blood film from a green iguana with two immature, mildly toxic heterophils and a monocyte. (Wright–Giemsa stain; original magnification X500). (c) Blood film from a green turtle with four immature, toxic heterophils. (Wright–Giemsa stain; original magnification X500). (d) Blood film from a caiman lizard with a markedly toxic heterophil. (Wright–Giemsa stain; original magnification X1000) © 2019 British Small Animal Veterinary Association

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8.18 (a) Blood film from a green iguana with moderately toxic heterophils. (Wright–Giemsa stain; original magnification X200). (b) Blood film from a green iguana with two immature, mildly toxic heterophils and a monocyte. (Wright–Giemsa stain; original magnification X500). (c) Blood film from a green turtle with four immature, toxic heterophils. (Wright–Giemsa stain; original magnification X500). (d) Blood film from a caiman lizard with a markedly toxic heterophil. (Wright–Giemsa stain; original magnification X1000)

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8.19

(a) Green turtle eosinophil. (Wright–Giemsa stain; original magnification X1000). (b) Blood film from a green iguana: E = green eosinophil; H = left-shifted heterophil; M = azurophilic monocyte. (Wright–Giemsa stain; original magnification X200) © 2019 British Small Animal Veterinary Association

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8.19 (a) Green turtle eosinophil. (Wright–Giemsa stain; original magnification X1000). (b) Blood film from a green iguana: E = green eosinophil; H = left-shifted heterophil; M = azurophilic monocyte. (Wright–Giemsa stain; original magnification X200)

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8.20

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8.20 Blood film from a rainbow boa: B = basophil; E = eosinophil; T = thrombocyte. (Wright–Giemsa stain; original magnification X500)

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8.21

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8.21 Blood film from a gopher tortoise: plasma cell. (Wright–Giemsa stain; original magnification X1000)

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8.22

Blood film from a pine snake: monocyte with two phagocytosed erythrocytes and a small amount of globular dark basophilic–black pigment suggestive of haemosiderin. (Wright–Giemsa stain; original magnification X200) © 2019 British Small Animal Veterinary Association

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8.22 Blood film from a pine snake: monocyte with two phagocytosed erythrocytes and a small amount of globular dark basophilic–black pigment suggestive of haemosiderin. (Wright–Giemsa stain; original magnification X200)

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8.24

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8.24 Fine-needle aspiration for cytology and bacterial culture/sensitivity testing in a day gecko. (Courtesy of Paul Raiti)

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8.25

Loggerhead sea turtle: direct smear of coelomic fluid with proteinaceous background, total protein of 3.5 g/dl, 35,000/μl WBC estimate (heterophils) consistent with exudate; infectious agents were absent. (Wright–Giemsa stain; original magnification X100) © 2019 British Small Animal Veterinary Association

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8.25 Loggerhead sea turtle: direct smear of coelomic fluid with proteinaceous background, total protein of 3.5 g/dl, 35,000/μl WBC estimate (heterophils) consistent with exudate; infectious agents were absent. (Wright–Giemsa stain; original magnification X100)

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8.26

Performing a lung wash in a green tree python with pneumonia. A sterile flexible catheter is placed through the glottis and into the distal trachea. (Courtesy of Paul Raiti) © 2019 British Small Animal Veterinary Association

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8.26 Performing a lung wash in a green tree python with pneumonia. A sterile flexible catheter is placed through the glottis and into the distal trachea. (Courtesy of Paul Raiti)

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8.27

(a) Crested gecko: Entamoeba invadens cyst in a faecal wet mount. (b) Crested gecko: cyst in a faecal wet mount stained with iodine. (c) Crested gecko: cyst in a faecal direct smear (trichrome stain). (Courtesy of Dr Heather Walden and Toni McIntosh, University of Florida, USA) © 2019 British Small Animal Veterinary Association

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8.27 (a) Crested gecko: Entamoeba invadens cyst in a faecal wet mount. (b) Crested gecko: cyst in a faecal wet mount stained with iodine. (c) Crested gecko: cyst in a faecal direct smear (trichrome stain). (Courtesy of Dr Heather Walden and Toni McIntosh, University of Florida, USA)

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8.28

(a) Gopher tortoise, swab of nasal discharge: heterophils and mononuclear inflammatory cells with phagocytosed bacilli. (Wright–Giemsa stain; original magnification X1000) (b) Gram-stained slide of the same preparation as in Figure 8.25 with presence of Gram-negative bacilli. (Original magnification X1000) (c) Loggerhead sea turtle: lung lesion imprint with histiocytic and heterophilic inflammation and branching septate hyphae. (Wright–Giemsa stain; original magnification X100) (d) Preparation of Figure 8.28c with GMS (Gomori’s methenamine silver) stain. (e) Gopher tortoise, nasal discharge direct smear: heterophilic inflammation and spirilliform bacteria. The bacteria stained positive with Warthin–Starry and were identified as Helicobacter sp. by PCR. (Wright–Giemsa stain; original magnification X100) (f) Bearded dragon: anaplastic sarcoma with mild heterophilic inflammation on eyelid. (Wright–Giemsa stain; original magnification X100) © 2019 British Small Animal Veterinary Association

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8.28 (a) Gopher tortoise, swab of nasal discharge: heterophils and mononuclear inflammatory cells with phagocytosed bacilli. (Wright–Giemsa stain; original magnification X1000) (b) Gram-stained slide of the same preparation as in Figure 8.25 with presence of Gram-negative bacilli. (Original magnification X1000) (c) Loggerhead sea turtle: lung lesion imprint with histiocytic and heterophilic inflammation and branching septate hyphae. (Wright–Giemsa stain; original magnification X100) (d) Preparation of Figure 8.28c with GMS (Gomori’s methenamine silver) stain. (e) Gopher tortoise, nasal discharge direct smear: heterophilic inflammation and spirilliform bacteria. The bacteria stained positive with Warthin–Starry and were identified as Helicobacter sp. by PCR. (Wright–Giemsa stain; original magnification X100) (f) Bearded dragon: anaplastic sarcoma with mild heterophilic inflammation on eyelid. (Wright–Giemsa stain; original magnification X100)

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8.29

Collecting a swab sample for bacterial culture and sensitivity testing from the capsule of a granuloma, after debridement, in a green iguana. (Courtesy of Paul Raiti) © 2019 British Small Animal Veterinary Association

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8.29 Collecting a swab sample for bacterial culture and sensitivity testing from the capsule of a granuloma, after debridement, in a green iguana. (Courtesy of Paul Raiti)