Broche Nelson, Takeshita Rafaela S C, Mouri Keiko, Bercovitch Fred B, Huffman Michael A
Primate Research Institute, Kyoto University, Inuyama, Aichi, Japan.
Universidade Federal Rural da Amazônia, Belém, Pará, Brazil.
Primates. 2019 Nov;60(6):547-558. doi: 10.1007/s10329-019-00757-6. Epub 2019 Sep 20.
Salivary alpha-amylase (sAA) enzyme functions as a digestive enzyme in many species that consume starch in their diet. Human studies have also revealed that sAA enzyme activity levels are positively correlated with the release of the stress hormone norepinephrine, allowing sAA to act as a biomarker for sympathetic nervous system activity. Recent non-human primate studies have incorporated sAA as a physiological stress marker. However, no published reports have investigated the time course of sAA from a stressful event to return to baseline levels in non-human primates. Furthermore, no validation of sAA as a stress biomarker has been reported for Japanese macaques (Macaca fuscata). This study had two primary aims: (1) to develop a systematic method for non-invasive saliva collection and, (2) to investigate sAA as a biomarker of acute stress in M. fuscata in order to better understand its acute stress-related characteristics. We developed a non-invasive method for cooperative saliva collection using positive reinforcement training (PRT) and tracked individual progress over 595 trials in ten individually housed Japanese macaques. We detected sAA enzyme in M. fuscata via kinetic reaction assay, then performed 22 acute stress tests. Four tests met conditions for interpreting sAA in response to an acute stressor and these results show that on average sAA activity rapidly increased post-stressor (mean ± SD = 4.2 ± 0.9 min) and returned to baseline shortly thereafter (10.4 ± 0.6 min). Our report reveals for the first time the temporal dynamics of sAA when applying acute stress to Japanese macaques and could be a useful tool for assessing animal welfare.
唾液α-淀粉酶（sAA）在许多以淀粉为食的物种中作为消化酶发挥作用。人体研究还表明，sAA酶活性水平与应激激素去甲肾上腺素的释放呈正相关，使得sAA能够作为交感神经系统活动的生物标志物。最近的非人灵长类动物研究已将sAA纳入生理应激标志物。然而，尚无已发表的报告研究过在非人灵长类动物中sAA从应激事件到恢复至基线水平的时间进程。此外，对于日本猕猴（Macaca fuscata），尚无关于sAA作为应激生物标志物的验证报告。本研究有两个主要目的：（1）开发一种用于非侵入性唾液采集的系统方法，以及（2）研究sAA作为日本猕猴急性应激的生物标志物，以便更好地了解其与急性应激相关的特征。我们开发了一种使用正强化训练（PRT）进行合作性唾液采集的非侵入性方法，并在十只单独饲养的日本猕猴的595次试验中跟踪个体进展。我们通过动力学反应测定法在日本猕猴中检测到了sAA酶，然后进行了22次急性应激测试。四项测试符合解释sAA对应激源反应的条件，这些结果表明，平均而言，sAA活性在应激源出现后迅速增加（平均值±标准差=4.2±0.9分钟），并在随后不久恢复至基线水平（10.4±0.6分钟）。我们的报告首次揭示了对日本猕猴施加急性应激时sAA的时间动态，并且可能成为评估动物福利的有用工具。
