Bricker Nelson K, Raskin Rose E, Densmore Christine L
Department of Comparative Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA.
Vet Clin Pathol. 2012 Sep;41(3):353-61. doi: 10.1111/j.1939-165X.2012.00452.x. Epub 2012 Aug 1.
Mechanisms of amphibian diseases are not characterized as well as those in domestic mammalian species. Antemortem laboratory testing is limited in frogs, presenting a diagnostic challenge to zoos, laboratories, and exotic veterinarians.
This study aimed to characterize blood cells and splenic cells from 2 anuran species based on characteristics identified by Wright staining, cytochemical staining, and immunochemical analysis and on histologic examination of spleens.
Blood specimens and spleens were obtained from 2 species of frog, the American bullfrog (Rana [Aquarana] catesbeiana) and the African clawed frog (Xenopus laevis). Blood smears were evaluated after Wright staining and cytochemical staining for α-naphthyl butyrate esterase (NBE), chloroacetate esterase (CAE), myeloperoxidase (PER), Sudan black B (SBB), and leukocyte alkaline phosphatase (LAP) reactions and for immunoreactivity for antibodies against CD3ε, CD79a, and BLA.36 antigens. Histologic sections of spleen were evaluated after staining with H&E and for immunoreactivity for CD3ε, CD79a, and BLA.36 antigens.
In bullfrogs, neutrophils, eosinophils, and monocytes were positive for some or all of the following: NBE, CAE, PER, and SBB; lymphocytes occasionally were positive for CAE. In clawed frogs, neutrophils, basophils, and monocytes were positive for some or all of the following: NBE, CAE, PER, and SBB; eosinophils occasionally were positive for CAE and PER, and lymphocytes were negative for all cytochemical stains. LAP was not a useful marker for any leukocyte type. In both species, peripheral blood lymphocytes were strongly immunoreactive for CD3ε, CD79a, and BLA.36. In splenic tissue, histologic patterns varied and there was diffuse immunoreactivity for CD79a and BLA.36 with focal reactivity for CD3ε, but with different distribution patterns in each species.
Cytochemical and immunochemical analysis of cells may be helpful in identification and characterization of amphibian blood cells and splenic cells for evaluation of the health of these animals.
两栖动物疾病的机制不像家养哺乳动物那样被充分了解。蛙类的生前实验室检测受限，这给动物园、实验室和外来动物兽医带来了诊断挑战。
本研究旨在根据瑞氏染色、细胞化学染色、免疫化学分析所确定的特征以及脾脏的组织学检查，对两种无尾两栖类动物的血细胞和脾细胞进行特征描述。
从两种蛙类，即美国牛蛙（牛蛙属[水生牛蛙亚属]牛蛙）和非洲爪蟾（非洲爪蟾）获取血液标本和脾脏。对血液涂片进行瑞氏染色和细胞化学染色，检测α-萘丁酸酯酶（NBE）、氯乙酸酯酶（CAE）、髓过氧化物酶（PER）、苏丹黑B（SBB）和白细胞碱性磷酸酶（LAP）反应，以及针对CD3ε、CD79a和BLA.36抗原的抗体免疫反应性。脾脏组织切片经苏木精-伊红染色后进行评估，并检测CD3ε、CD79a和BLA.36抗原的免疫反应性。
在牛蛙中，中性粒细胞、嗜酸性粒细胞和单核细胞对以下部分或全部检测呈阳性：NBE、CAE、PER和SBB；淋巴细胞偶尔对CAE呈阳性。在爪蟾中，中性粒细胞、嗜碱性粒细胞和单核细胞对以下部分或全部检测呈阳性：NBE、CAE、PER和SBB；嗜酸性粒细胞偶尔对CAE和PER呈阳性，淋巴细胞对所有细胞化学染色均呈阴性。LAP对任何白细胞类型都不是一个有用的标志物。在这两个物种中，外周血淋巴细胞对CD3ε、CD79a和BLA.36均有强烈的免疫反应性。在脾脏组织中，组织学模式各不相同，CD79a和BLA.36呈弥漫性免疫反应性，CD3ε呈局灶性反应性，但在每个物种中的分布模式不同。
细胞的细胞化学和免疫化学分析可能有助于识别和描述两栖动物的血细胞和脾细胞，以评估这些动物的健康状况。
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