Hematologic reference intervals for koi (Cyprinus carpio), including blood cell morphology, cytochemistry, and ultrastructure

BACKGROUND: Hematologic data are used routinely in the health care of humans and domestic mammals. Similar data for fish are largely fragmentary or have not been collected. OBJECTIVES: The primary purpose of this study was to determine hematologic reference intervals for koi, an ornamental strain of the common carp (Cyprinus carpio). Secondarily, the morphology, cytochemical reactions, and ultrastructure of koi blood cells were characterized. METHODS: A CBC was performed manually on heparin-anticoagulated blood specimens using Natt and Herrick's diluent and a Neubauer-ruled hemacytometer. Leukocyte differential counts were done on Wright-Leishman- and Diff-Quik-stained blood smears. Cytochemical reactions of koi leukocytes were determined using commercial kits. Transmission electron microscopy was performed to characterize the ultrastructural features of koi blood cells. RESULTS: Hematologic reference intervals were established for healthy koi for PCV (30-34%), hemoglobin concentration (6.3-7.6 g/dL), RBC count (1.7-1.9 X 10(6)/ microL), WBC count (19.8-28.1 X 10(3)/ microL), RBC indices, and differential leukocyte counts. Lymphocytes were the predominant leukocyte (accounting for up to 80% of all leukocytes), whereas eosinophils were rare. Basophils were positive with PAS staining. Naphthol AS-D chloroacetate esterase activity was observed only in eosinophils. alpha-Naphthyl butyrate esterase and beta-glucuronidase activities were positive in monocytes. Some lymphocytes were reactive for alpha-naphthyl butyrate esterase and acid phosphatase activity. Ultrastructurally, leukocytes, erythrocytes, and thrombocytes were identified on the basis of cytoplasmic organelles and granule appearance. CONCLUSION: Hematologic reference intervals and knowledge of the cytochemical reactions and ultrastructural characteristics of koi leukocytes will help standardize hematologic studies in this species.     

Morphologic and cytochemical characteristics of blood cells of juvenile loggerhead sea turtles (Caretta caretta)

A morphologic classification based on the cytochemical characteristics of blood cells of 35 juvenile loggerhead sea turtles (Caretta caretta) is described. Cytochemical stains included benzidine peroxidase, chloroacetate esterase, alpha-naphthyl butyrate esterase (with and without sodium fluoride), acid phosphatase (with and without tartaric acid), Sudan black B, periodic acid-Schiff, and toluidine blue. The morphologic characteristics of erythrocytes were similar to those reported in green turtles. Six types of white blood cells were identified: heterophils, eosinophils, basophils, lymphocytes, monocytes and thrombocytes. Except for the basophils, the rest of the white blood cells from loggerhead turtles had different cytochemical characteristics compared to blood cells from other sea turtle species. The leukocyte differential count was different from that reported for other sea turtle species. Heterophils were the most numerous leukocytes from these loggerhead turtles, followed by lymphocytes, eosinophils, monocytes and basophils. This paper provides a morphologic classification of blood cells of loggerhead sea turtles that is useful for veterinary surgeons involved in sea turtle conservation.     

Hematology and plasma chemistry reference intervals for cultured shortnose sturgeon (Acipenser brevirostrum)

BACKGROUND: The shortnose sturgeon, Acipenser brevirostrum, is an imperiled species distributed along the Atlantic coast of North America. Interest in replenishing wild stocks with hatchery-reared fish has created a need for accurate hematologic and biochemical reference intervals to evaluate the health of both fish raised in aquaculture systems and fish in the wild. OBJECTIVES: The objective of this study was to generate hematologic and biochemistry reference intervals for healthy shortnose sturgeon. METHODS: Blood samples were collected in heparinized tubes from 77 shortnose sturgeon raised in flow-through aquaculture systems. Whole blood and plasma samples were analyzed for hematologic and biochemical variables using standard techniques. Reference intervals were calculated as the central 95% (percentile) of data. RESULTS: Hematologic reference intervals (n = 46) were as follows: PCV 26-46%, hemoglobin 5.7-8.7 g/dL, MCV 307-520 fL, MCH 65.9-107.1 pg, MCHC 15-30 g/dL, plasma proteins (refractometry) 2.8-6.0 g/dL, RBC count 0.65-1.09 x 10(6)/microL, total WBC count 28,376-90,789/microL, small lymphocytes 9063-56,656/microL, large lymphocytes 2122-10,435/microL, neutrophils 3758-33,592/microL, monocytes 0-7137/microL, eosinophils 0-1544/microL, thrombocyte-like cells 6863-23,046/microL, thrombocytes 32,205-122,179/microL, and neutrophil:lymphocyte ratio 0.068-1.026. Plasma chemistry reference intervals (n = 77) were as follows: total protein 2.7-5.3 g/dL, albumin 0.8-1.7 g/dL, globulins 1.8-3.7 mg/dL, creatinine 0-1.4 mg/dL, total bilirubin 0-0.1 mg/dL, alkaline phosphatase 47-497 U/L, aspartate aminotransferase 90-311 U/L, sodium 124-141 mmol/L, potassium 2.9-3.7 mmol/L, chloride 106-121 mmol/L, calcium 6.6-12.1 mg/dL, magnesium 1.6-2.3 mg/dL, phosphorus 5.1-8.1 mg/dL, glucose 37-74 mg/dL, cholesterol 42-133 mg/dL, and osmolality 232-289 mOsm/kg. CONCLUSION: Reference values reported here will be useful for the early detection, identification, and monitoring of disease and sublethal conditions in cultured shortnose sturgeon.     

A light and electron microscopic study of the leukocytes of channel catfish (Ictalurus punctatus)

A light and electron microscopic study of channel catfish (Ictalurus punctatus) white blood cells was carried out. The white blood cells of channel catfish consist of lymphocytes, neutrophils, thrombocytes and monocytes. A fifth cell type is illustrated and described which could not be characterized. These leukocytes are characterized and compared with their mammalian counterparts.     

Morphological and Cytochemical Studies of Peripheral Blood Cells of Schizothorax prenanti

The morphological and cytochemical studies of peripheral blood cells of Schizothorax prenanti were studied by light and electron microscopy. Erythrocytes, thrombocytes and three types of leucocytes, lymphocytes, neutrophils and monocytes, were distinguished and characterized. In addition to mature erythrocytes, immature and dividing erythrocytes were observed. A few organelles such as mitochondria were distributed in the cytoplasm of erythrocytes. Lymphocytes with heavily clumped heterochromatic nucleus and minimal cytoplasm were classified into small and large lymphocytes. Three different populations of granules, with distinctive ultrastructural aspect, were observed in neutrophils. Monocytes were the fewest leucocytes possessing rich organelles, phagocytized materials and vacuoles. Thrombocytes with various types were the most abundant blood cells among leucocytes and contained a prominent nucleus with dense bands of heterochromatin and many cytoplasmic vacuoles. Periodic acid‐Schiff staining was positive in neutrophils, monocytes, lymphocytes and thrombocytes, but not in erythrocytes. Peroxidase‐positive staining was observed in neutrophils and monocytes, but not in erythrocytes, lymphocytes and thrombocytes. Only neutrophils were positive for oil red O. Except for erythrocytes, the other blood cells stained positively for acid phosphatase. Only neutrophils and monocytes were positive for α‐naphthyl acetate esterase. None of the cells studied were positive for alkaline phosphatase. The morphologic and cytochemical features of blood cells of S. prenanti are similar to those of other fish. This investigation may be helpful as a tool to monitor the health status of cultured S. prenanti and will grant early detection of clinical pathology.     

Hematology and clinical chemistry of adult yellow-headed temple turtles (Hieremys annandalii) in Thailand

Background: Yellow‐headed temple turtles (YHT), Hieremys annandalii, native to Thailand, are protected from exploitation under the Wild Animal Reservation and Protection Act, also listed under Appendix II of the Convention on International Trade of Endangered Species and the International Union for the Conservation of Nature red list. Objectives: The objectives of this study were to describe quantitative, morphologic, and cytochemical features of blood cells and plasma biochemical analytes of clinically healthy YHT. Methods: Blood samples were collected from 40 adult YHT from October 2007 to February 2008. Hematologic and biochemical analyses, cytochemical staining, and ultrastructural evaluation were performed using standard methods. Results: Hematologic results (mean ± SD) included: RBC count, 0.275 ± .094 × 10⁶ cells/μL; WBC count, 11.7 ± 6.6 × 10³ cells/μL; heterophils, 29.4 ± 6.9%; eosinophils, 23.7 ± 5.3%; basophils, 21.2 ± 1.9%; lymphocytes, 14.8 ± 5.9%; and azurophils, 10.7 ± 5.3%. Erythrocytes stained dark red with peroxidase‐staining. Periodic acid‐Schiff stain could not differentiate between thrombocytes and lymphocytes. Thrombocytes contained cytoplasmic vacuoles, similar to mammalian platelets and those of birds and snakes. Heterophils and eosinophils were similar in structure and cytochemical staining characteristics to those of other turtles and reptiles. Structure of basophils was similar to avian basophils. Lymphocytes and azurophils had similar cytochemical staining compared with mammalian lymphocytes and monocytes. Mean MCHC, WBC counts, absolute azurophil counts, and plasma alanine aminotransferase activity were higher in male turtles than in females. Conclusion: Blood characteristics of YHT are species‐specific, and this study can be served as a reference for future clinical studies and medical care of YHT.     

Haematological values of post-laying Arrau turtle (Podocnemis expansa) in the Orinoco River, Venezuela

The Arrau turtle (Podocnemis expansa) is an endangered species, as a result of long-lasting, unsustainable exploitation. To obtain reference haematological values from the wild Podocnemis expansa during post-laying, 20 turtles were captured in the Orinoco River. Blood was obtained from the dorsal cervical sinus in lithium heparin tubes. Red blood cells (RBC), white blood cells (WBC), thrombocytes (TC), packed cell volume (PCV), plasmatic protein (PP), haemoglobin (Hgb), mean corpuscular volume (MCV) and differential leukocyte count were determined. Haematological values were: RBC 0.9×10(9)/L, WBC 5.7×10(9)/L, TC 5.4×10(9)/L, PCV 35.6%, PP 4.2g/dL, Hgb 11.8g/dL, MCV 411fL. The differential leukocyte count comprised: 71% heterophils, 23% lymphocytes, 3% eosinophils, 1.6% basophils, and 1% monocytes. The reports of reference haematology values for the wild P. expansa are limited; therefore, the results presented herein contrast with those values obtained in captivity. This study represents a contribution to the referential haematological values of the wild P. expansa.     

Characterization of blood cells in the Leopard Cat (Prionailurus bengalensis)

Background: The Leopard Cat (Prionailurus bengalensis) is the most frequently encountered wild cat in most of Southeast Asia. Limited hematologic investigation exists for this species. Objectives: The objectives of this study were to assess routine hematologic measurements and parameters and characterize the morphology, cytochemical staining, and ultrastructural features of blood cells in Leopard Cats. Methods: Blood samples were collected from 12 adult healthy captive Leopard Cats (7 males and 5 females). Complete blood counts were performed using an automated hematology analyzer and manual differential counts. Cytochemical staining (Sudan black B [SBB], peroxidase [PO], periodic acid‐Schiff [PAS], α‐naphthyl acetate esterase [ANAE], and β‐glucuronidase [BG]) and scanning and transmission electron microscopy were performed using standard methods. Results: Median (range) hematologic results were as follows: PCV 0.46 L/L (0.30–0.55 L/L), hemoglobin 136.5 g/L (100–183 g/L), WBC 9.0 × 10⁹/L (6.9–15.2 × 10⁹/L), band neutrophils 0.07 × 10⁹/L (0–0.30 × 10⁹/L), segmented neutrophils 2.9 × 10⁹/L (1.2–6.34 × 10⁹/L), lymphocytes 5.3 × 10⁹/L (2.7–8.1 × 10⁹/L), eosinophils 0.14 × 10⁹/L (0–0.73 × 10⁹/L), basophils 0/L (0–0.22 × 10⁹/L), and monocytes 0.08 × 10⁹/L (0–0.30 × 10⁹/L). Neutrophils stained strongly positive for SBB, PO, and PAS; lymphocytes had fine granular positivity for ANAE and BG; monocytes were weakly positive for ANAE and BG; and basophils were strongly positive for BG. Ultrastructurally, eosinophils contained many large rod‐shaped granules with prominent crystalloid core structures, ribosomes, and mitochondria. Basophils contained many round to oval specific granules with homogeneous contents. Low number of basophils contained a few small vacuoles that usually were not detected by light microscopy. Conclusion: These findings will facilitate interpretation of hematologic results for future investigative and diagnostic studies of this species.     

Ultrastructural and Cytochemical Properties of Peripheral Blood Cells of Piebald Naked Carp (Gymnocypris eckloni)

The ultrastructural and cytochemical properties of peripheral blood cells of Gymnocypris eckloni were investigated by transmission electron microscopy and a range of cytochemical techniques to provide clear insight into the structure and function of blood cells from this fish. Ultrastructurally, erythrocytes, leucocytes (neutrophils, eosinophils, lymphocytes, monocytes), thrombocytes and plasma cells were identified in the peripheral blood of G. eckloni. The most special ultrastructural characteristics of blood cells in this fish were that neutrophils exhibited only one type of cytoplasmic granules containing an eccentric, spherical or oval electron‐dense core, and eosinophils presented two types of granules with non‐uniform electronic density and without crystalloids in their cytoplasm. Neutrophils, eosinophils, lymphocytes, monocytes and thrombocytes were positive for periodic acid–Schiff and α‐naphthyl acetate esterase staining. Intense peroxidase positive staining was observed in neutrophils and monocytes, but not in eosinophils, lymphocytes and thrombocytes. Neutrophils, eosinophils and monocytes were stained positively for acid phosphatase, whereas lymphocytes and thrombocytes did not stain. Leucocytes and thrombocytes were negative for alkaline phosphatase and Sudan black B staining. Erythrocytes were negative for all cytochemical staining. The cytochemical and ultrastructural features of peripheral blood cells of G. eckloni were similar to those of other fish species. However, some important differences were identified in G. eckloni.     

Changes in Peripheral Blood Leukocyte Percentages and Function of Neutrophils in Stressed Channel Catfish

Abstract

Channel catfish Ictalurus punctatus were subjected to a simulated handling (including bleeding) and transport stress. Using flow cytometry in conjunction with monoclonal antibodies to channel catfish B and T lymphocytes (minimal cross-reaction with thrombocytes) and neutrophils coupled with routine hematological methods, we demonstrated significant decreases in the percentage of B lymphocytes and increases in T lymphocytes and neutrophils of transported fish, yet there was no change in neutrophil phagocytic function. These data suggest that the length of the stressing event or the duration of exposure of the neutrophils to cortisol could determine if immune responses become suppressed. These data prompted us to expose channel catfish in vivo, and isolated neutrophils in vitro, to cortisol at various concentrations and for various time periods. Exposure of neutrophils to cortisol concentrations of 100 μg/dL for 2 h did not suppress phagocytosis; however, significant decreases in the percent phagocytosis and bacterial killing and increases in total number of neutrophils isolated were obtained after repeated in vivo administration of cortisol. The results of these experiments indicate (1) that bleeding, handling, and transport of channel catfish induce a characteristic relative lymphopenia and neutrophilia, (2) that cortisol does not act alone to induce suppression of phagocytic function (based on the in vitro administration of cortisol), and (3) that high physiological concentration of cortisol in vivo can initiate phagocytic suppression. In general, the intraperitoneal cortisol injection experiments suggested that stress on channel catfish that produces a concomitant high serum concentration of cortisol can suppress neutrophil phagocytic function, perhaps providing an avenue for the onset of infection.     

Differential cytochemical staining characteristics of channel catfish leukocytes identify cell populations in lymphoid organs

This is one of the first characterizations of channel catfish (Ictalurus punctatus) leukocytes by enzyme cytochemistry. Leukocytes demonstrated cytoplasmic staining patterns very similar to mammalian leukocytes when stained with acid phosphatase, alpha-naphthyl butyrate esterase, beta-glucuronidase, alpha-naphthyl acetate esterase, Sudan Black B and anti-immunoglobulin specific immunohistochemistry. Lymphocytes, monocytes, macrophages, neutrophils, and surface immunoglobulin positive (surface Ig+) cells were present in channel catfish renal hematopoietic tissue and spleen and demonstrated distinctive cytoplasmic foci staining patterns, cytoplasmic blushing or cell membrane staining. Monocytes, macrophages, lymphocytes and surface Ig+ cells were present in the thymus. Thymic and splenic cellular organization appeared very similar to these same mammalian tissues. In the thymus, acid phosphatase positive cells were distributed throughout the parenchyma, while alpha-naphthyl butyrate esterase and beta-glucuronidase positive cells were concentrated in the cortex and the medulla, respectively. Surface immunoglobulin positive cells occurred in the cortex. In the spleen, acid phosphatase positive cells were scattered throughout the parenchyma, while alpha-naphthyl butyrate esterase positive cells were scattered throughout the parenchyma and adjacent to splenic arterioles. Beta-glucuronidase and surface immunoglobulin positive cells were restricted to immediately adjacent to splenic arterioles. Sudan Black B positive cells were scattered throughout the parenchyma, while alpha-naphthyl acetate esterase positive cells occurred adjacent to peri-arteriole lymphoid sheaths and appear very similar to mammalian metallophils.     

Hematologic characteristics of captive western barred bandicoots (Perameles bougainville) from Western Australia

BACKGROUND: The western barred bandicoot (Perameles bougainville) is an Australian marsupial species now considered endangered as a consequence of habitat destruction and predation. A recently discovered papillomatosis syndrome is hindering efforts to repopulate this species. Hematology reference intervals have been lacking for P bougainville, preventing optimal interpretation of hematology results from wart-affected and clinically normal animals. OBJECTIVES: The purpose of this study was to establish hematology reference values and describe morphologic characteristics of blood cells of healthy western barred bandicoots. METHODS: Fifty-nine whole blood samples were collected by jugular venipuncture into EDTA from 47 clinically healthy captive western barred bandicoots at 3 locations on the Western Australian mainland. A CBC was performed using an ADVIA-120 analyzer. Data were compared on the basis of geographic location, sex, age, and lactation status, and reference intervals were calculated. Blood cell morphology was evaluated using light microscopy, and transmission and scanning electron microscopy. RESULTS: Significant differences were found based on sex (RBC indices, fibrinogen), age (% polychromatophilic RBCs), and geographic location (RBC, neutrophil, and lymphocyte counts, MCHC, % polychromatophilic RBCs, fibrinogen). Combined reference intervals were calculated for hemoglobin concentration (122-165 g/L), HCT (0.36-0.49 L/L), and total WBC (2.9-14.9 x 10(9)/L), monocyte (0-0.6 x 10(9)/L), eosinophil (0-0.9 x 10(9)/L), and total plasma protein (47-63 g/L) concentrations. Leukocyte, erythrocyte, and platelet morphology were similar to those of other marsupial peramelid species. Nuclei in neutrophils, monocytes, and eosinophils occasionally had an annular configuration. CONCLUSIONS: Reference intervals and blood cell morphology obtained in this study will be useful for the evaluation of laboratory data from ill animals and assist with population health monitoring of western barred bandicoots.     

Evaluation of white blood cell concentration,plasma fibrinogen concentration,and an agar gel immunodiffusion test for early identification of foals with Rhodococcus equi pneumonia

OBJECTIVE: To evaluate WBC concentration, plasma fibrinogen concentration, and an agar gel immunodiffusion (AGID) test for early identification of Rhodococcus equi-infected foals. DESIGN: Prospective study. ANIMALS: 162 foals from a farm with enzootic R equi infection. PROCEDURE: Blood samples were obtained from each foal at 4-week intervals for measurement of WBC and plasma fibrinogen concentrations and at 2-week intervals for detection of anti-R equi antibody by an AGID assay. Diagnostic performance of WBC and fibrinogen concentrations was assessed by use of receiver operating characteristic curve analysis. For each assay, sensitivity, specificity, and predictive values were calculated at various cutoff points; bacteriologic culture of R equi from a tracheobronchial aspirate was used as the reference standard test. RESULTS: Diagnostic performance of WBC concentration was significantly higher than that of fibrinogen concentration. Sensitivity and specificity of measurement of WBC concentration at a cutoff of 13,000 cells/microL were 95.2 and 61.2%, respectively; at a cutoff of 15,000 cells/microL, sensitivity was 78.6% and specificity was 90.8%. When a positive test result was used as the cutoff, sensitivity of the AGID assay was 62.5% and specificity was 53.8%. CONCLUSION AND CLINICAL RELEVANCE: Monitoring WBC concentration is a useful approach for early detection of infected foals on farms with a high prevalence of R equi pneumonia. In contrast, serologic surveillance by use of an AGID assay is of little benefit for that purpose.     

Morphology, cytochemical staining, and ultrastructural characteristics of the blood cells of the giant lizard of El Hierro (Gallotia simonyi)

The object of this study was to examine the erythrocytes, leukocytes and thrombocytes of the giant lizard of El Hierro (Gallotia simonyi) by light and electron (TEM) microscopy, and cytochemical staining. Smears were prepared from blood from the ventral coccygeal vein of 10 healthy adult lizards (five males and five females) from the Giant Lizard of El Hierro Reproduction and Research Centre, Canary Islands, Spain. The cytochemical stains used were: benzidine peroxidase (BP), chloroacetate esterase (CAE), alpha-naphthyl acetate esterase (ANAE), acid phosphatase (AP), periodic acid-Schiff (PAS), toluidine blue (TB) and May-Grünwald-Giemsa (MGG). Electron microscopy was also performed on all samples. Heterophils had granules that were heterogeneous in both size and electron density, and stained with BP, PAS and ANAE. Eosinophil granules were homogeneously electron-dense and stained for AP, CAE and ANAE. Basophils had both highly and moderately electron-dense granules, and stained with TB and ANAE. Azurophil granules were of low electron-density and stained for AP, CAE and ANAE. Azurophil cytoplasm was vacuolated on TEM. The cytoplasm of lymphocytes contained many ribosomes and was positive for AP. Monocytes had a large nucleus and a vacuolated cytoplasm but did not stain by any of the cytochemical methods used. Thrombocytes had a relatively large nucleus but little cytoplasm; they did not stain cytochemically. The blood cells of the giant lizards of El Hierro differ from those of other members of the Order Squamata both morphologically and cytochemically. The variation in cytochemical responses in the blood of reptiles makes it necessary to study species individually if meaningful clinical decisions are to be made.     

Immune Response and Resistance of Channel Catfish to Edwardsiella ictaluri Challenge when Fed Various Dietary Levels of Zinc Methionine and Zinc Sulfate

Abstract

The effects of dietary levels of zinc from zinc methionine (ZnM) and zinc sulfate ZnSO₄ on nonspecific immune responses and disease resistance of juvenile channel catfish Ictalurus punctatus (mean initial weight of 3.3 g) were determined. Fish in triplicate aquaria were fed to satiation twice daily for 16 weeks with an egg white basal diet without zinc supplementation or supplemented with 5, 20, and 60 mg zinc/kg as ZnM or 20 and 60 mg zinc/kg as ZnSO₄. Relative percentages of peripheral blood lymphocytes, thrombocytes, and macrophages were not affected by dietary zinc. The relative percentages of neutrophils significantly increased in fish fed 20 and 60 mg Zn/kg as ZnM. Chemotactic responses of macrophages were significantly higher for fish fed the three ZnM-supplemented diets and the 60-mg ZnSO₄. Dietary zinc, however, had no influence on the phagocytic activity of macrophages for Edwardsiella ictaluri. Likewise, neither the source nor the level of dietary zinc provided protection to channel catfish against E. ictaluri. The severity of E. ictaluri infection (number of colony-forming units/g trunk kidney) and the percentage offish that were culture negative for E. ictaluri (15 d postchallenge) were also unaffected by dietary zinc.     

Morphology,cytochemical staining and ultrastructural characteristics of reindeer (Rangifer tarandus) leukocytes

Peripheral blood smears from four adult reindeer (Rangifer tarandus) were examined after staining with Romanowsky's stain and cytochemical stains, including alpha-napthyl butyrate esterase (alpha-NBE), Sudan black B (SBB), chloroacetate esterase (CAE) and alkaline phosphatase (ALP). Romanowsky-stained eosinophils, neutrophils, lymphocytes and monocytes resembled those of cattle, sheep and goats. Basophils had two different staining patterns with Romanowsky's stain. Basophils that we termed "grey basophils" were similar in appearance to grey eosinophils in Greyhound dogs, with medium blue-grey to lavender-grey cytoplasm containing varying numbers of clear vacuoles or granules and variable numbers of small, intensely basophilic, perinuclear granules. The second basophil staining pattern was more typical of ruminant basophils, with uniform, pale to dark basophilic cytoplasmic granules. Basophils stained positive for alpha-NBE, SBB, CAE, and ALP. Eosinophils stained positive for SBB, and were negative for alpha-NBE, CAE, and ALP. Neutrophils were negative for SBB, CAE, and ALP. Monocytes stained positive for alpha-NBE, were rarely positive for CAE and SBB, and were negative for ALP. Transmission electron microscopy revealed matrix within all granulocytes granules, including those of basophils.     

Hematology, leukocyte cytochemical analysis, plasma biochemistry, and plasma electrophoresis of wild-caught and captive-bred Gila monsters (Heloderma suspectum)

Background: The Gila monster (Heloderma suspectum), one of several venomous lizard species in the world, is found within the United States and Mexico and is recognized as an iconic symbol of the American Southwest. Thus, Gila monsters are of growing interest in the captive reptile trade and within zoological and educational institutions. Objectives: The aims of this study were to determine results for CBCs, describe cytochemical reactions in WBCs, and obtain plasma biochemical and protein electrophoresis results from wild‐caught and captive‐bred H. suspectum. Methods: Ventral tail vein blood samples were obtained from 16 captive (14 wild‐caught and 2 captive‐bred) Gila monsters. CBCs, RBC morphometric analysis, plasma biochemical analysis, and protein electrophoresis were performed. Leukocytes were stained for peroxidase, Sudan black B (SBB), chloroacetate esterase, napthyl butyrate esterase, and leukocyte alkaline phosphatase, and blood smears were examined for the presence of hemoparasites. Results: The median (range) PCV was 37% (22–50%) and WBC count was 4.6 × 10³/μL (3.3–6.4 × 10³/μL) with approximately 50% heterophils and fewer lymphocytes, basophils, azurophils, and monocytes in decreasing order. Cytochemical reactions were unique among reptiles with strong staining for peroxidase and SBB in monocytes/azurophils. Biochemical results were similar to those of earlier reports with slight increases in uric acid and urea concentrations. Plasma electrophoretic results indicated that albumin was approximately equal to the combined globulin fractions. Conclusions: Results of blood analysis in healthy wild‐caught and captive‐bred H. suspectum may be useful for monitoring health status in this species.     

Morphologic, cytochemical staining, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes (Crotalus adamanteus)

OBJECTIVE: To evaluate light microscopic, cytochemical, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes. ANIMALS: 10 healthy snakes. PROCEDURE: Various stains, including Wright-Giemsa, benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff with diastase, and toluidine blue, were used to stain leukocytes differentially on multiple blood smears. Electron microscopy also was performed. RESULTS: Lymphocytes were the most commonly observed leukocyte and could be distinguished from thrombocytes, using periodic acid-Schiff stain with diastase. Azurophils also were commonly observed; their granules stained with peroxidase. Eosinophils were not identified; however, 2 morphologic variations of heterophils were seen in the blood of all snakes and were considered the same cell type at different stages of cytoplasmic granule development. Heterophil granules were better preserved, using a one-step Wright-Giemsa method that did not require alcohol fixation prior to staining. Degranulated heterophils were observed in all preparations. CONCLUSIONS: Most leukocytes of eastern diamondback rattlesnakes can be identified easily on Wright-Giemsa-stained preparations. However, hematologic stains that do not require alcohol fixing prior to staining may be preferred for leukocyte evaluation in certain reptiles. A limited degree of heterophil maturation may continue in the blood of healthy snakes. This, along with degranulation of heterophils, may result in a variable staining pattern in this cell type, regardless of the stain used. CLINICAL RELEVANCE: Results provide baseline data for use in hematologic testing in diagnosis of disease and monitoring of treatment of sick or injured snakes.     

Chemical reactions in cells from leukemic cats

Cytochemical staining for leukocyte alkaline phosphatase(LAP), nonspecific esterase (NSE), nonspecific esterase with fluoride inhibition (NSE-F), periodic acid Schiff (PAS) reactivity, and peroxidase (PO) was valuable in identification of the neoplastic cell type in 10 leukemic cats. Staining both blood and bone marrow smears was often necessary for making the correct diagnosis. Cytochemical staining resulted in changing the morphologic diagnosis of leukemia in two of the 10 cats. Also, increased LAP activity, probably a marker for myelocytic leukemia in the cat, was observed in bone marrow cells from three nonleukemic, FeLV-positive cats.