Validation of an Enzyme-Linked Immunosorbent Assay and Establishment of a Reference Interval for Androstenedione in Texas Tortoises (Gopherus berlandieri)

Garrett C. Crooks, Jacquelyn K. Grace, Jianhua Guo, Alice Blue-McLendon, J. Jill Heatley
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Abstract

This study aimed to validate an enzyme-linked immunosorbent assay (ELISA) and create a reference interval for androstenedione from captive Texas tortoises (Gopherus berlandieri). Reference interval creation for androstenedione could allow comparison of an individual's values to others of the species in an effort to evaluate excess or deficiency and/or provide the basis for diagnosis of reproductive health or capability. In other species, androstenedione is associated with puberty onset and specific reproductive diseases; however, values and function of this hormone remain poorly studied in reptiles. The 25 adult male tortoises sampled were maintained at Texas A&M University's Winnie Carter Wildlife and Exotic Animal Center. Serum was assayed via a validated androstenedione ELISA. Male Texas tortoise serum androstenedione concentrations were found to be 5.40 ± 5.16 ng/ml (mean ± SD), with establishment of a reference interval of 0.57–18.58 ng/ml. Relatively increased androstenedione values were associated with increased carapace length, carapace height, and tortoise weight but not gular horn length of male Texas tortoises. This information may benefit the care, management, and reproduction of captive tortoises. Extrapolation should be considered for evaluation of other species until their own intervals are established. Further research should include determination of baseline values of androstenedione for both healthy and reproductively challenged female tortoises.
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酶联免疫吸附法的验证及德州象雄烯二酮参考区间的建立
本研究旨在验证酶联免疫吸附试验(ELISA),并建立圈养德克萨斯象龟雄烯二酮的参考区间。建立雄烯二酮的参考区间可以将个体值与该物种的其他个体值进行比较,以评估过量或不足和/或为诊断生殖健康或能力提供基础。在其他物种中,雄烯二酮与青春期发病和特定生殖疾病有关;然而,这种激素的价值和功能在爬行动物中的研究仍然很少。这25只成年雄性陆龟被保存在德克萨斯农工大学的温妮·卡特野生动物和外来动物中心。经验证的雄烯二酮酶联免疫吸附试验检测血清。雄性德州龟血清雄烯二酮浓度为5.40±5.16 ng/ml (mean±SD),建立了0.57 ~ 18.58 ng/ml的参考区间。雄甾二酮值的相对增加与雄性德州象甲壳长度、甲壳高度和龟重的增加有关,但与正常角长度无关。这些信息可能有利于圈养陆龟的护理、管理和繁殖。对其他物种的评价应考虑外推,直到它们自己的间隔确定为止。进一步的研究应包括确定健康和生殖障碍雌龟雄烯二酮的基线值。
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