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in Journal of the American Veterinary Medical Association

SUMMARY

In these studies, the effects of recombinant human interleukin 2 (rHuil-2) were examined on in vitro tumor cytotoxicity by canine blood lymphocytes obtained from peripheral vessels through use of a chromium release microcytotoxicity assay. Cytotoxic activity by lymphokine-activated killer cells was significantly increased, compared with that by untreated lymphocytes in a dose-dependent manner. The maximal effect was attained with 300,000 IU of rHuil-2/ml. Lymphokine-activated killing also was dependent on the duration of incubation with rHuil-2. After 1 day of rHuil-2 incubation, cytotoxicity was significantly increased, compared with that of untreated lymphocytes. Of the 3 times examined, cytotoxicity peaked after 3 days of rHuil-2 incubation. High levels of cytotoxic activity were still present at 7 days of incubation. Numbers of granular lymphocytes increased over the times examined. These results demonstrate functional and morphologic changes in canine peripheral blood lymphocytes obtained from peripheral vessels after incubation with rHuil-2 in a dose- and time-dependent manner.

Free access
in American Journal of Veterinary Research

Abstract

Objective—To report values for tear production, central corneal touch threshold (CTT), and intraocular pressure (IOP) in healthy guinea pigs and determine results of aerobic bacterial culture and cytologic examination of conjunctival swab specimens.

Design—Cross-sectional study.

Animals—31 healthy guinea pigs (62 eyes) of various ages and breeds.

Procedures—Tear production was measured by the phenol red thread tear test (PRT) and Schirmer tear test (STT) before and after topical anesthetic application, CTT was measured with an esthesiometer, and IOP was measured by applanation tonometry.

Results—Combining data from all eyes, mean ± SD PRT values before and after topical anesthetic administration were 21.26 ± 4.19 mm/15 s and 22.47 ± 3.31 mm/15 s, respectively, and mean IOP was 18.27 ± 4.55 mm Hg. Median STT values before and after topical anesthetic administration were 3 mm/min (range, 0 to 12 mm/min) and 4 mm/min (range, 0 to 11 mm/min), respectively, and median CTT was 2.0 cm (range, 0.5 to 3.0 cm). Values did not differ between eyes for any test, but significant differences were identified for PRT values between males and females and between values obtained before and after topical anesthetic administration. Common bacterial isolates included Corynebacterium spp, Streptococcus spp, and Staphylococcus spp. Cytologic examination of conjunctival swab specimens revealed mainly basal epithelial cells; lymphocytes were common.

Conclusions and Clinical Relevance—Results provided information on values for PRT, STT, CTT, and IOP in healthy guinea pigs and on expected findings for aerobic bacterial culture and cytologic examination of conjunctival swab specimens.

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in Journal of the American Veterinary Medical Association

Abstract

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, α-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 diamond-back 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. (Am J Vet Res 1999;60:507-514)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To identify and characterize blood cells from free-ranging Hawaiian green turtles, Chelonia mydas.

Sample Population

26 green turtles from Puako on the island of Hawaii and Kaneohe Bay on the island of Oahu.

Procedure

Blood was examined, using light and electron microscopy and cytochemical stains that included benzidine peroxidase, chloroacetate esterase, alpha naphthyl butyrate esterase, acid phosphatase, Sudan black B, periodic acid-Schiff, and toluidine blue.

Results

6 types of WBC were identified: lymphocytes, monocytes, thrombocytes, heterophils, basophils, and eosinophils (small and large). Morphologic characteristics of mononuclear cells and most granulocytes were similar to those of cells from other reptiles except that green turtles have both large and small eosinophils.

Conclusions

Our classification of green turtle blood cells clarifies improper nomenclature reported previously and provides a reference for future hematologic studies in this species. (Am J Vet Res 1998;59:1252–1257)

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in American Journal of Veterinary Research

SUMMARY

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, α-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, α-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.

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in American Journal of Veterinary Research

Abstract

Objectives

To determine optimal site for collection of bone marrow from desert tortoises, and to characterize cytologic staining and morphologic features of bone marrow hematopoietic cells.

Animals

16 desert tortoises.

Procedure

Bone marrow was obtained at necropsy from the pelvis, proximal portion of the humerus, femur, and thickened portions of the cranial to craniolateral and caudal to caudolateral margins of the carapace and plastron for histologic and cytologic examinations. Cytocentrifuged preparations of marrow cells were evaluated for reactivity to cytochemical stains.

Results

Histologic sections were adequate for evaluating acidophils, acidophil precursors, and erythrocyte precursors. It was difficult to differentiate among monocytes, lymphocytes, thrombocytes, and blast cells, and eosinophils could not be differentiated from heterophils. Basophils were in rare, small clusters of 3 to 12 cells. A few lymphoid follicles were found in the pelvis and long bones.

Use of cytochemical staining accomplished differentiation between agranular heterophil precursors and granulated heterophils, and between granulated eosinophils and basophils. Monocytes, azurophils, and monoblasts had similar staining features. Staining of erythrocyte precursors with Sudan black B differentiated them from lymphocytes. Only a few small cells with periodic acid-Schiff-positive cytoplasm were identified as thrombocytes. Lymphocytes did not stain with any of the cytochemical stains.

Conclusions

For histologic and cytologic evaluation of bone marrow hematopoietic cells, pelvis, proximal portion of the humerus, femur, and thickened portions of the peripheral cranial and caudal regions of the carapace and plastron are suitable sites to collect specimens. There are distinct cytochemical markers for heterophil, monocyte, and erythrocyte precursors, as well as later stage heterophils, eosinophils, basophils, monocytes, and azurophils. (Am J Vet Res 1996;57:1608–1615)

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in American Journal of Veterinary Research