Validation of an enzyme immunoassay for measurement of fecal dehydroepiandrosterone sulfate in gibbons and siamangs
Validation of an enzyme immunoassay for measurement of fecal dehydroepiandrosterone sulfate in gibbons and siamangs
复制标题
长臂猿和合趾猴粪便硫酸脱氢表雄酮测量酶免疫分析的验证
DOI：
10.1002/zoo.21687
复制
发表时间：
2022
期刊：
影响因子：
1.3
通讯作者：
Takeshita, Rafaela S.
中科院分区：
文献类型：
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作者：
Takeshita, Rafaela S.
监测野生动物的压力水平对于确保它们在圈养或野外的生活质量至关重要。测定糖皮质激素(GC)和脱氢表雄酮硫酸盐(DHEAS)是评估应激反应的一种有前景的方法，这两种肾上腺激素参与应激反应的调节。虽然非侵入性方法测量GC已经在几个物种中得到验证，但只有几项研究验证了DHEAS分析。这项研究的目的是(1)描述一种测定DHEAS水平的酶免疫测定法，(2)对长臂猿和暹曼的粪便样本进行验证，以及(3)测试一次冻融循环后以及在两种冷冻温度(−20°C和−80°C)下的激素稳定性。研究对象包括来自美国动物园的32只长臂猿和暹罗猿猴。EIA通过并行性和准确性测试进行分析验证，并通过三个个体在应激事件(事故、疫苗接种或交通)后1-2天确认DHEAS反应进行生物学验证。此外，怀孕女性的粪便DHEAS水平高于非妊娠/非哺乳期水平，并在分娩后逐渐下降。激素稳定性实验显示，在一个冻融周期后，粪便DHEAS水平没有明显变化。无论储存温度如何，粪便提取物中的激素水平在2个月内都是稳定的，在−20℃和−80℃条件下没有显著差异。该方法灵敏度高，适用于长臂猿、暹罗等动物粪便中DHEAS的测定，具有推广应用的潜力。
Monitoring wildlife stress levels is essential to ensure their quality of life in captivity or in the wild. One promising method to assess the stress response is the comeasurement of glucocorticoids (GC) and dehydroepiandrosterone sulfate (DHEAS), adrenal hormones involved in the modulation of the stress response. Although noninvasive methods to measure GCs have been validated in several species, only a few studies have validated DHEAS assays. The aims of this study were (1) to describe an enzyme immunoassay (EIA) to measure DHEAS levels, (2) to validate this assay for fecal samples in gibbons and siamangs, and (3) to test hormonal stability after one freeze‐thaw cycle and over time at two freezer temperatures (−20°C and −80°C). Subjects included 32 gibbons and siamangs from U.S. zoological parks. The EIA was validated analytically by parallelism and accuracy tests, and biologically by confirming a DHEAS response 1–2 days after a stressful event (accident, vaccination, or transportation) in three individuals. In addition, fecal DHEAS levels in a pregnant female were above nonpregnant/nonlactating levels and declined progressively the following parturition. The hormonal stability experiments revealed no significant changes in fecal DHEAS levels after one freeze‐thaw cycle. Hormonal levels in fecal extracts were stable for 2 months, regardless of the storage temperature, with no significant differences between −20°C and −80°C conditions. The EIA described has high sensitivity and it is suitable for fecal DHEAS measurement in gibbons and siamangs, with a potential to be applied to other species.