Large carnivores play critical roles in the maintenance and function of natural ecosystems; however, the populations of many of these species are in decline across the globe. Therefore, there is an urgent need to develop novel techniques that can be used as sensitive conservation tools to detect new threats to the health of individual animals well in advance of population-level effects. Our study aimed to determine the expression of proteins related to energetics, reproduction and stress in the skin of grizzly bears (Ursus arctos) using a liquid chromatography and multiple reaction monitoring mass spectrometry assay. We hypothesized that a suite of target proteins could be measured using this technique and that the expression of these proteins would be associated with biological (sex, age, sample location on body) and environmental (geographic area, season, sample year) variables. Small skin biopsies were collected from free-ranging grizzly bears in Alberta, Canada, from 2013 to 2019 (n = 136 samples from 111 individuals). Over 700 proteins were detected in the skin of grizzly bears, 19 of which were chosen as targets because of their established roles in physiological function. Generalized linear mixed model analysis was used for each target protein. Results indicate that sample year influenced the majority of proteins, suggesting that physiological changes may be driven in part by responses to changes in the environment. Season influenced the expression of proteins related to energetics, reproduction and stress, all of which were lower during fall compared to early spring. The expression of proteins related to energetics and stress varied by geographic area, while the majority of proteins that were affected by biological attributes (age class, sex and age class by sex interaction) were related to reproduction and stress. This study provides a novel method by which scientists and managers can further assess and monitor physiological function in wildlife.

中文翻译：

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灰熊健康蛋白质生物标志物的开发和验证。

大型食肉动物在自然生态系统的维护和功能中起着至关重要的作用。但是，全球许多此类物种的数量都在下降。因此，迫切需要开发可用作敏感保护工具的新技术，以在种群影响之前就检测出对个体动物健康的新威胁。我们的研究旨在通过液相色谱和多反应监测质谱法确定与灰熊（Ursus arctos）皮肤能量，繁殖和应激相关的蛋白质表达。我们假设可以使用此技术测量一组目标蛋白，并且这些蛋白的表达与生物学（性别，年龄，样本在人体上的位置）和环境（地理区域，季节，样本年）变量。2013年至2019年，从加拿大艾伯塔省的自由放养的灰熊收集了小型皮肤活检样本（n =来自111个个体的136个样本）。在灰熊的皮肤中检测到700多种蛋白质，由于它们在生理功能中已确立的作用，因此选择了19种作为目标。每种靶蛋白均使用广义线性混合模型分析。结果表明样本年份影响了大多数蛋白质，表明生理变化可能部分是由对环境变化的响应所驱动。季节影响了与能量，生殖和压力有关的蛋白质的表达，与早期的春季相比，秋季的所有这些蛋白质含量都较低。与能量和压力有关的蛋白质表达随地理区域而异，而受生物学特性（年龄，性别和受性别相互作用影响的年龄类别）影响的大多数蛋白质与生殖和压力有关。这项研究提供了一种新颖的方法，科学家和管理人员可以通过该方法进一步评估和监测野生动植物的生理功能。