As studies quantifying steroid hormones in marine mammal blubber progress, methodological refinements may improve the utility and consistency of blubber hormone measurements. This study advances blubber extraction methodologies by testing a simplified extraction protocol that reduces time and complexity compared to a protocol widely used in cetacean blubber studies. Using blubber samples archived from remote biopsy (n = 21 live whales) and necropsy collection (n = 7 dead whales) of North Atlantic right whales (NARW; Eubalaena glacialis) of known life history states, we performed analytical and biological validations to assess the feasibility of measuring reproductive (testosterone, progesterone) and glucocorticoid (cortisol) hormones in blubber via enzyme immunoassay following the simplified extraction. Analytical validations (parallelism, accuracy, extraction efficiency, repeatability) showed the simplified extraction produced similar results to the extended protocol, offering a more efficient and consistent technique. In live, apparently healthy whales, blubber testosterone concentrations (mean ± SE) were significantly higher in males (2.02 ± 0.36 ng/g) compared to females (0.81 ± 0.15 ng/g). Blubber progesterone was highest in a confirmed pregnant female (60.3 ng/g), which was 12-fold greater than the mean concentration of non-pregnant females (4.56 ± 0.88 ng/g). Blubber cortisol concentrations in whales that died from anthropogenic causes averaged 5.31 ± 2.28 ng/g, whereas most live, healthy whales had cortisol values below 1 ng/g. Among living whales, a whale actively entangled in fishing gear had the highest blubber cortisol measurement (3.51 ng/g), exhibiting levels similar to whales that died from acute entanglement (2.88 ± 0.42 ng/g). Overall, the highest blubber cortisol concentration (18.0 ng/g) was measured in a dead whale with a severe chronic entanglement, approximately 30-fold greater than mean blubber cortisol of apparently healthy whales (0.58 ± 0.11 ng/g). The methodological approach presented here provides a reference for researchers interested in an alternative, streamlined technique for hormone extraction of cetacean blubber and contributes to the diverse tool set for stress and reproductive assessments of endangered NARWs.

中文翻译：

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使用简化的激素提取方案在北大西洋右鲸脂中检测到压力和生殖事件

随着定量研究海洋哺乳动物油脂中类固醇激素的研究进展，方法的改进可能会改善油脂激素测量的效用和一致性。这项研究通过测试简化的提取协议来改进油脂提取方法，该协议相比于鲸类鲸脂研究中广泛使用的协议减少了时间和复杂性。使用从远程活检（n  = 21的活鲸）和尸检收集（n  = 7死的鲸）收集的北大西洋右鲸（NARW; Eubalaena glacialis）的脂肪样本）的已知生命史状态，我们进行了分析和生物学验证，以评估简化提取后通过酶免疫法测定啤酒脂中生殖（睾丸激素，孕酮）和糖皮质激素（皮质醇）激素的可行性。分析验证（并行性，准确性，提取效率，可重复性）表明，简化的提取产生了与扩展方案相似的结果，从而提供了一种更有效，更一致的技术。在活的，显然健康的鲸鱼中，雄性（2.02±0.36 ng / g）的雄性比雄性（0.81±0.15 ng / g）的雄性睾丸激素浓度显着更高。确诊的孕妇中的黄体孕酮最高（60.3 ng / g），比未怀孕女性的平均浓度（4.56±0.88 ng / g）高12倍。死于人为原因的鲸鱼中的皮质醇平均含量为5.31±2.28 ng / g，而大多数活的健康鲸鱼的皮质醇值均低于1 ng / g。在活的鲸鱼中，活跃地缠在渔具中的鲸脂的鲸脂皮质醇测量值最高（3.51 ng / g），表现出与因急性缠结而死的鲸鱼相似的水平（2.88±0.42 ng / g）。总的来说，在有严重慢性缠结的死鲸中，最高脂糖皮质醇浓度（18.0 ng / g）被测量，比明显健康的鲸鱼的平均脂糖皮质醇（0.58±0.11 ng / g）高约30倍。本文介绍的方法论方法为感兴趣的替代方法的研究人员提供了参考，