Cattet Marc, Stenhouse Gordon B, Janz David M, Kapronczai Luciene, Anne Erlenbach Joy, Jansen Heiko T, Nelson O Lynne, Robbins Charles T, Boulanger John
RGL Recovery Wildlife Health & Veterinary Services, 415 Mount Allison Crescent, Saskatoon, Saskatchewan, CanadaS7H 4A6.
Department of Veterinary Pathology, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan, CanadaS7N 5B4.
Conserv Physiol. 2017 Jun 1;5(1):cox032. doi: 10.1093/conphys/cox032. eCollection 2017.
Recognizing the potential value of steroid hormone measurements to augment non-invasive genetic sampling, we developed procedures based on enzyme-linked immunoassays to quantify reproductive steroid hormone concentrations in brown bear () hair. Then, using 94 hair samples collected from eight captive adult bears over a 2-year period, we evaluated (i) associations between hair concentrations of testosterone, progesterone, estradiol and cortisol; (ii) the effect of collecting by shaving vs. plucking; and (iii) the utility of reproductive hormone profiles to differentiate sex and reproductive state. Sample requirements (125 mg of guard hair) to assay all hormones exceeded amounts typically obtained by non-invasive sampling. Thus, broad application of this approach will require modification of non-invasive techniques to collect larger samples, use of mixed (guard and undercoat) hair samples and/or application of more sensitive laboratory procedures. Concentrations of hormones were highly correlated suggesting their sequestration in hair reflects underlying physiological processes. Marked changes in hair hormone levels during the quiescent phase of the hair cycle, coupled with the finding that progesterone concentrations, and their association with testosterone levels, differed markedly between plucked and shaved hair samples, suggests steroids sequestered in hair were likely derived from various sources, including skin. Changes in hair hormone concentrations over time, and in conjunction with key reproductive events, were similar to what has been reported concerning hormonal changes in the blood serum of brown bears. Thus, potential for the measurement of hair reproductive hormone levels to augment non-invasive genetic sampling appears compelling. Nonetheless, we are conducting additional validation studies on hair collected from free-ranging bears, representative of all sex, age and reproductive classes, to fully evaluate the utility of this approach for brown bear conservation and research.
认识到类固醇激素测量对于增强非侵入性基因采样的潜在价值,我们开发了基于酶联免疫测定的程序,以量化棕熊( )毛发中的生殖类固醇激素浓度。然后,我们使用在两年时间内从八只圈养成年熊身上采集的94份毛发样本,评估了(i)睾酮、孕酮、雌二醇和皮质醇的毛发浓度之间的关联;(ii)剃毛与拔毛采集方式的影响;以及(iii)生殖激素谱在区分性别和生殖状态方面的效用。检测所有激素所需的样本量(125毫克保护毛)超过了非侵入性采样通常获得的量。因此,要广泛应用这种方法,需要改进非侵入性技术以采集更大的样本,使用混合(保护毛和绒毛)毛发样本和/或应用更灵敏的实验室程序。激素浓度高度相关,表明它们在毛发中的封存反映了潜在的生理过程。毛发周期静止期毛发激素水平的显著变化,再加上孕酮浓度及其与睾酮水平的关联在拔毛和剃毛样本之间存在显著差异,表明封存于毛发中的类固醇可能来自包括皮肤在内的各种来源。毛发激素浓度随时间的变化以及与关键生殖事件的关联,与有关棕熊血清激素变化的报道相似。因此,测量毛发生殖激素水平以增强非侵入性基因采样的潜力似乎很有吸引力。尽管如此,我们正在对从自由放养的熊身上采集的毛发进行额外的验证研究,这些熊代表了所有性别、年龄和生殖类别,以全面评估这种方法在棕熊保护和研究中的效用。