Leukocyte and thrombocyte reference values for channel catfish (Ictalurus punctatus Raf), with an assessment of morphologic, cytochemical, and ultrastructural features

BACKGROUND: Hematology tests are useful to evaluate physiologic disturbances in fish and can provide important information for the diagnosis and prognosis of disease. OBJECTIVES: The primary purpose of this study was to define reference intervals for thrombocytes and leukocytes in healthy channel catfish (Ictalurus punctatus). In addition, the morphologic, cytochemical, and ultrastructural features of blood cells were assessed. METHODS: Blood samples (0.5 mL) were collected into EDTA from 40 clinically healthy catfish on a commercial fish farm in Jaboticabal, Brazil. Thrombocyte, total WBC, and differential WBC counts were determined and reference intervals were calculated as the 25-95th percentiles of data. Thrombocyte and leukocyte morphology was assessed in blood smears stained with May Grünwald-Giemsa-Wright and ultrastructurally by transmission electron microscopy. Cytochemical staining patterns were described using periodic acid-Schiff (PAS), peroxidase, nonspecific esterase, alkaline phosphatase, and toluidine blue. RESULTS: Reference intervals were as follows: thrombocytes 58,802-99,569/microL; total WBCs 27,460-41,523/microL; lymphocytes 5380-11,581/microL; monocytes 2949-7459/microL; neutrophils 12,529-22,748/microL, and basophils 736-2003/microL. Neutrophils were positive for peroxidase and PAS; monocytes were positive for nonspecific esterase; and basophils were positive with toluidine blue. CONCLUSION: The morphologic and staining features of neutrophils and monocytes of channel catfish are similar to those of mammals, and the presence of basophils in this species was verified. These reference intervals and morphologic findings provide a foundation for future investigations on the functions and alterations of blood cells in channel catfish.     

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.     

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.     

Hematologic values in calves during the first 6 months of life

BACKGROUND: Age-related changes in hematologic values are known to occur in many species. Few published studies include repeated measurements of hematologic parameters in calves during the first months of life. OBJECTIVE: The aim of the present study was to monitor hematologic values by sequential measurements from birth to 6 months of age in 15 healthy calves of the Norwegian Red breed, and compare the results to reference intervals for adult, lactating dairy cows. METHODS: Fifteen clinically healthy calves were sampled every week during the first 5 weeks of life and every month thereafter until 6 months of age. Hematologic values were measured using the ADVIA 120 hematology system. Reference intervals were determined for 75 healthy adult cows of the same breed. RESULTS: Compared with adult reference intervals, the MCV was lower and the RBC count was higher in calves throughout the investigation period. Hemoglobin concentration stayed largely within the adult reference interval. Mean MCHC was lower than adult values for 5 weeks, then increased and reached adult values by weeks 10-12. The mean lymphocyte count for calves reached adult reference values at weeks 6-8, and the mean monocyte count increased steadily until weeks 14-16. For most leukocytes, interindividual variation was larger during the first 5-8 weeks of life. The mean platelet count for calves was higher than the adult reference interval until weeks 19-21 of age. CONCLUSIONS: Age-specific reference intervals for calves from birth to 6 month of age are needed for RBC count, MCV, MCHC, red cell distribution width, and platelet and lymphocyte counts.     

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.     

Morphologic characterization of specific granules in Greyhound eosinophils

BACKGROUND: "Vacuolated" eosinophils (ie, eosinophils with empty, nonstaining granules) have been described previously in normal Greyhounds. However, to our knowledge, detailed studies of granules in vacuolated and normal eosinophils in this breed have not been performed. OBJECTIVE: The objective of this prospective study was to characterize some of the morphologic, ultrastructural, and cytochemical staining features of specific (primary) granules in both normal and vacuolated eosinophils in Greyhound blood. METHODS: Morphologic features of eosinophils in Wright's- and Diff-Quik-stained peripheral blood smears from 49 Greyhounds were compared with 200 blood smears from non-Greyhound dogs. Transmission electron microscopy was done on blood from 3 Greyhounds with vacuolated eosinophils and 3 with normal eosinophil granules. Blood smears from 4 of these dogs also were stained cytochemically with alkaline phosphatase (AP), chloracetate esterase (CAE), and alpha naphthyl butyrate esterase (ANBE). The morphologic features and tinctorial properties of vacuolated and normal eosinophils were compared. RESULTS: Twenty-six Greyhounds (53%) had vacuolated eosinophils and 23 (47%) had normal granulated eosinophils in smears stained with Wright's stain. Only 1% of eosinophils were vacuolated in non-Greyhound dogs. Twenty of the 23 (85%) Greyhounds with normal granulated eosinophils on Wright's-stained smears had vacuolated eosinophils in smears stained with Diff-Quik. Ultrastructurally, no morphologic differences were observed between granules of vacuolated and normal eosinophils. Both vacuolated and normal eosinophils in Greyhounds were positive for AP and negative for CAE and ANBE, as expected for normal dogs. CONCLUSION: Vacuolated eosinophils in Greyhounds likely reflect, at least in part, differential staining properties of the specific granules with different hematologic stains. Ultrastuctural and cytochemical features of eosinophil granules were similar in normal and vacuolated eosinophils from Greyhounds.     

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.     

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.     

Flow cytometric patterns in blood from dogs with non-neoplastic and neoplastic hematologic diseases using double labeling for CD18 and CD45

BACKGROUND: In dogs, flow cytometry is used in the phenotyping of immunologic cells and in the diagnosis of hemic neoplasia. However, the paucity of specific antibodies for myeloid cells and B lymphocytes and of labeled antibodies for multicolor techniques limits the ability to detect all leukocyte subpopulations. This is especially true for neoplastic and precursor cells. CD18 and CD45 are expressed on all leukocytes and are involved in cell activation, and together could be useful in helping determine cell lineage. OBJECTIVES: The purpose of this study was to double label canine blood for CD18 and CD45 and to use the differential expression of antigens to identify leukocyte populations in dogs with non-neoplastic and neoplastic hematologic diseases. METHODS: A template was developed using blood samples from 10 clinically healthy dogs and a back-gating technique. Differential leukocyte counts obtained with the template were compared with those obtained by manual and automated methods on blood samples from 17 additional healthy dogs. Blood samples obtained from 9 dogs with non-neoplastic (reactive) hematologic diseases and 27 dogs with hemic neoplasia were double stained for CD18 and CD45 using mouse anticanine CD18 monoclonal antibody (mAb) plus phycoerythrin-conjugated rat anticanine CD45 mAb and fluorescein isothiocyanate-conjugated rabbit antimouse IgG. Hemic neoplasms were diagnosed by cell morphology, and immunophenotypic and cytochemical markers. RESULTS: With the double label, neutrophils, eosinophils, monocytes, and T- and B-lymphocytes were identified. In reactive disorders, a population of activated neutrophils with high CD45 and CD18 expression was detected. In hemic neoplasia, cell lineage was easily determined, even in acute leukemia. CONCLUSIONS: Double labeling for CD18/CD45 may be useful as a screening method to evaluate hematologic diseases and help determine cell lineage, and to aid in the selection of a panel of antibodies that would be useful for further analysis.     

Hematology of the sandbar shark, Carcharhinus plumbeus: standardization of complete blood count techniques for elasmobranchs

BACKGROUND: Standardized hematologic methods and reference intervals have not been established for cartilaginous fishes (sharks, skates, and rays) despite the large number of animals displayed in zoos and aquariums worldwide. OBJECTIVE: The focus of this study was to validate CBC methods for sandbar shark (Carcharhinus plumbeus) blood, based on criteria established in human medicine, for the following tests: RBC count, total WBC count, PCV, hemoglobin (Hgb) concentration, and WBC differential percentages. METHODS: Replicate CBCs were performed using blood samples from 5 captive sandbar sharks. Three protocols for RBC and total WBC counts were compared, as were different centrifugation times for PCV determination, and 2 methods for Hgb concentration. Means, minimum and maximum values, and CVs were compared to CAP and CLIA performance guidelines for human tests. RESULTS: Total WBC counts in a diluent modified for elasmobranch blood, Hgb concentration by the cyanmethemoglobin method after removal of nuclei, and WBC differential percentages showed acceptable performance. PCV results were acceptable when tubes were centrifuged for at least 5 minutes. Total RBC counts by all 3 methods exceeded the acceptable error for manual counts of human cells. CONCLUSIONS: Standardized CBC tests can be used as health assessment tools for elasmobranchs. Total RBC counts should be viewed as estimates.     

Grey eosinophils in a Miniature Schnauzer with a poorly differentiated mast cell tumor

A 10‐year‐old female spayed Miniature Schnauzer was presented for investigation of an intra‐nasal mass. The mass was diagnosed by histopathologic examination as an undifferentiated round cell neoplasm with an infiltrate of segmented leukocytes, interpreted as neutrophilic inflammation. The mass was treated with palliative radiation and systemic chemotherapy due to the presence of regional lymph node metastasis. During subsequent monitoring over several months, the peripheral leukocyte concentration was repeatedly within reference intervals to slightly increased with low numbers of toxic neutrophils. Four months after the initial diagnosis, there was a significant leukocytosis of 66 100 cells/μL, and 39 700 cells/μL of the leukocytes had variably mature, lobulated, and hypolobulated nuclei, and grey cytoplasm with clear vacuoles, resembling grey eosinophils. To further characterize these cells, peripheral blood smears from the patient and a canine control with eosinophilia were stained for alkaline phosphatase (AP), peroxidase, and esterase activities, and with Luxol fast blue (LFB). Histopathologic sections of the nasal mass were stained with LFB and immunohistochemically for tryptase. On blood smears, the cytoplasm of the suspected grey eosinophils stained for AP and granules stained with LFB confirmed that there was an eosinophilic lineage. Peroxidase staining was weak, and esterase staining was absent. On histopathologic sections from the nasal mass, the segmented leukocytes contained LFB‐staining granules, indicating an eosinophilic infiltrate was present. Neoplastic cells expressed tryptase, which confirms a mast cell lineage. Our findings suggest that grey eosinophils might be under‐recognized and interpreted incorrectly as toxic neutrophils. This report expands the canine breeds in which these eosinophils have been identified.     

Hematologic, cytochemical, ultrastructural, and molecular findings of Hepatozoon-infected flat-headed cats (Prionailurus planiceps)

BACKGROUND: The flat-headed cat (Prionailurus planiceps) is a small wild cat of Southeast Asia and is considered extremely endangered. Little is known about the hematologic values, blood cell morphology, or hemoparasites of this species in relation to other Felidae. OBJECTIVES: The objective of this study was to report basic hematologic values and describe the light microscopic, cytochemical, and ultrastructural characteristics of blood cells in 2 wild-caught flat-headed cats. In addition, molecular analysis was done of a Hepatozoon organism found in the neutrophils of both cats. METHODS: Blood samples were collected into EDTA from the cephalic vein. A CBC, manual differential count, manual reticulocyte count, cytochemical stains (Sudan black B [SBB], alpha-naphthyl acetate esterase [ANAE], and beta-glucuronidase), and scanning and transmission electron microscopy were done using standard methods. RESULTS: HCT was slightly lower and reticulocyte counts and red cell distribution width were higher than the expected values for other species of cats. Hepatozoon organisms were found in the cytoplasm of neutrophils in both cats, but the number of infected neutrophils was very low (1%-2%). Neutrophils stained strongly positive for SBB, but were negative for ANAE and beta-glucuronidase. Hepatozoon-infected neutrophils were negative for SBB, but focally positive for ANAE and beta-glucuronidase. By transmission electron microscopy, gamonts of Hepatozoon sp were observed in neutrophils, and rarely free in plasma. Infected neutrophils had fewer specific granules and more mitochondria compared with noninfected neutrophils. PCR products of partial 18S rRNA revealed that the isolate of Hepatozoon in the flat-headed cats was closely related to that of the frog Hepatozoon sp. CONCLUSIONS: These results add to our understanding of hematologic values and blood cell morphology in Hepatozoon-infected flat-headed cats as well as the molecular analysis of the Hepatozoon organism, and may be useful for the health management and evaluation of hemoparasitic disease in this species.     

Morphologic and cytochemical characteristics of blood cells from the desert tortoise (Gopherus agassizii).

Morphologic and cytochemical staining characteristics of erythrocytes, leukocytes, and thrombocytes of the desert tortoise (Gopherus agassizii) were evaluated, using blood smears prepared from 23 healthy tortoises of Kern County, Calif. Special emphasis was placed on differentiating features of the various leukocytes and thrombocytes. A variety of cytochemical stains, including benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff, and toluidine blue were used. Heterophils had a characteristic, large, focal area of positive staining with chloroacetate esterase, alpha-naphthyl butyrate esterase, and acid phosphatase. Eosinophils stained diffusely positive with benzidine peroxidase, allowing differentiation of this leukocyte from heterophils. Thrombocytes stained focally positive with periodic acid-Schiff, allowing differentiation of these cells from lymphocytes, which stained uniformly negative. An intracytoplasmic body, commonly observed within erythrocytes, was considered ultrastructurally to represent a degenerate organelle.     

Myelogenous leukemia in a bearded dragon (Acanthodraco vitticeps).

A 3-yr-old bearded dragon (Acanthodraco vitticeps) presented with lethargy, a swollen right elbow joint, inability to move its rear limbs normally, and marked leukocytosis. The majority of leukocytes were an abnormal mononuclear lymphoid-type cell with a high nuclear to cytoplasmic ratio, a slightly blue cytoplasm, nuclei with coarsely granular chromatin, and some nuclear clefts. Acute leukemia of lymphoid or myeloid origin was tentatively diagnosed. The abnormal mononuclear leukocyte cell population stained positively for the myeloid cytochemical stains: peroxidase, chloroacetate esterase, and L1-calprotectin. The abnormal cell population of the peripheral blood did not stain with the lymphoid cytochemical stains: alpha-naphthyl butyrate esterase, CD3, and CD79a.     

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.     

Stomatocytosis in 7 related Standard Schnauzers

BACKGROUND: Hereditary canine stomatocytosis has been described in purebred Alaskan Malamutes, Drentse Patrijshonds, and Miniature Schnauzers. In humans, hereditary stomatocytosis is a heterogeneous group of congenital disorders characterized by the presence of stomatocytes in blood, increased osmotic fragility, and frequently, hemolytic anemia. OBJECTIVE: Our objective was to describe hematologic findings and RBC characteristics in 7 closely related Standard Schnauzers with stomatocytosis. METHODS: The following parameters were measured using an automated analyzer: HCT, RBC, hemoglobin (Hb) concentration, MCV, MCH, MCHC, red cell distribution width (RDW), WBC, platelet count, mean platelet volume (MPV), thrombocrit (PCT), and platelet distribution width (PDW). Differential leukocyte count, platelet estimate, reticulocyte count, and the percentage of stomatocytes in blood films were microscopically evaluated. An osmotic fragility test of RBCs and measurement of intracellular Na+, K+, and 2,3-diphosphoglycerate (2,3-DPG) concentrations were also performed. RESULTS: The affected dogs had macrocytosis (80.0 +/- 4.2 fL, reference interval 60-76 fL), decreased MCHC (29.3 +/- 0.8 g/dL, reference interval 32-39 g/dL), slightly increased RDW (17.3 +/- 0.4%, reference interval 12-16%), and an increased reticulocyte count (1.55 +/- 0.77%, reference interval <1%). The percentage of stomatocytes in blood films varied from 0.6 to 18.9% of all RBCs. Erythrocyte osmotic fragility and intracellular Na+ (138.1 +/- 3.2 mmol/L; controls 99 +/- 6.1 mmol/L), K+ (8.1 +/- 0.8 mmol/L; controls 6.1 +/- 0.5 mmol/L), and 2,3-DPG (21.9 +/- 2.0 micromol/g Hb; controls: 14.6 +/- 3.3 micromol/g Hb) concentrations were increased in dogs with stomatocytosis. CONCLUSIONS: Hematologic findings and the metabolic defects in RBCs in these Standard Schnauzers were consistent with a diagnosis of stomatocytosis. Parentage analysis suggests that stomatocytosis in Standard Schnauzers may have a hereditary component.     

Age-related changes in hematology and plasma chemistry values of hybrid striped bass (Morone chrysops X Morone saxatilis)

Hybrid striped bass (Morone chrysops X Morone saxatilis ) are an important aquaculture species yet there are few diagnostic tools available to assess their health. Hematology and clinical chemistry analyses are not used extensively in fish medicine due to the lack of reference intervals for various fish species, and because factors such as age can affect blood values. There is little published information regarding age-related changes in blood values of juvenile fish. It is important to evaluate juvenile fish, as this is the time they are raised in aquaculture settings. Determining age-related changes in the blood values of fishes would further develop clinical pathology as a diagnostic tool, enhancing both fish medicine and the aquaculture industry. The results of standard hematology and clinical chemistry analysis were evaluated in juvenile hybrid striped bass at 4, 6, 9, 15, and 19 months of age. Values for PCV and RBC indices were significantly lower, and plasma protein concentration was significantly higher in younger fish. Total WBC and lymphocyte counts were significantly higher in fish at 6 and 9 months of age, while neutrophil and monocyte counts were higher at 6, 9, and 15 months. Eosinophil counts were significantly higher in 9-month-old fish. The majority of hematologic values fell within previously established reference intervals, indicating that only slight modification to the intervals is necessary for evaluating hematologic results of hybrid striped bass at different ages. The following analytes deviated sufficiently from adult reference intervals to warrant separate reference values: plasma protein concentration at 4 months, WBC and lymphocyte counts at 15 and 19 months, and thrombocyte-like-cells at 9 months of age. Values for most biochemical analytes were significantly different among age groups except for creatinine and potassium concentrations. Comparisons with reference intervals were not made for biochemical analytes, because established reference intervals were not available. Age-related changes in hematologic and biochemical values of striped bass were similar to those reported for rainbow trout and mammals.     

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.     

Hematology of equine fetuses with comparisons to their dams

The Hematologic values of 19 equine fetuses between 202 and 238 days gestation were compared with those of their dams. The red blood cell (RBC) count, hemoglobin concentration, hematocrit, and mean corpuscular hemoglobin concentration were significantly lower in fetal blood, while the mean corpuscular volume, mean corpuscular hemoglobin, and red cell distribution width were significantly higher. Mares had a significantly higher nucleated blood cell count than fetuses, and all nucleated cells were leukocytes (WBC). Most WBC in mare blood were segmented neutrophils and lymphocytes. In contrast, over one-half of the nucleated cells in fetal blood were nucleated RBC, and the majority of WBC in fetal blood were lymphocytes. Mares also had significantly higher plasma protein and fibrinogen concentrations than their fetuses. Mild macrocytosis and mild polychromasia were observed in most fetal blood samples, but not in blood samples from mares. All fetal blood contained reticulocytes, and most samples contained Heinz bodies and Howell-Jolly bodies. The results of this study will contribute to the development of hematologic reference values that may be useful in equine fetal research and, possibly, in the diagnosis of equine fetal disease.     

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.