Research Institute of Wildlife Ecology, Department of Interdisciplinary Life Sciences, University of Veterinary Medicine, Vienna, Vienna, Austria; Unit of Veterinary Public Health and Epidemiology, Institute of Food Safety, Food Technology and Veterinary Public Health, Department for Farm Animals and Veterinary Public Health, University of Veterinary Medicine, Vienna, Vienna, Austria.
Konrad Lorenz Institute of Ethology, Department of Interdisciplinary Life Sciences, University of Veterinary Medicine, Vienna, Vienna, Austria; Department of Evolutionary Zoology and Human Biology, University of Debrecen, Debrecen, Hungary.
Physiol Behav. 2021 Sep 1;238:113488. doi: 10.1016/j.physbeh.2021.113488. Epub 2021 Jun 10.
Despite the widely used application of standardized capture-handling protocols to collect blood and assess the physiological stress response, the actual sampling design (e.g., timing and the number of blood samples) often differs between studies, and the potential implications for the measured physiological endpoints remain understudied. We, therefore experimentally tested the effects of repeated handling and multiple blood sampling on the stress response in wintering free-living great tits (Parus major). We modified a well-established sampling protocol of avian studies by adding either an additional blood sample or a "sham-manipulation" (i.e., handling associated with the blood sampling procedure without venepuncture), to disentangle the effects of handling stress and blood loss. We combined three different stress metrics along the endocrine-immune interface to investigate the acute short-term stress response: total corticosterone concentrations (Cort), the heterophil/lymphocyte ratio (H:L), and the Leucocyte Coping Capacity (LCC). Our study provided three key results: i) no relationship between Cort levels, LCC and H:L, confirming that these three parameters represent different physiological endpoints within the stress response; ii) contrasting dynamics in response to stress by the measured parameters and iii) no difference in physiological stress levels 30 min after capture due to one additional blood sampling or handling event. By optimising the sampling design, our results provide implications for animal welfare and planning experimental procedures on stress physiology in passerine species.
尽管标准化的采集和处理方案被广泛应用于采集血液并评估生理应激反应，但在不同的研究中，实际的采样设计（例如，采样时间和样本数量）往往存在差异，而这对测量生理终点的潜在影响仍未得到充分研究。因此，我们通过实验测试了重复处理和多次采血对冬季自由生活的大山雀（Parus major）应激反应的影响。我们通过添加额外的血液样本或“假操作”（即与采血过程相关但不进行静脉穿刺的操作）来修改鸟类研究中已建立的采样方案，以区分处理应激和失血的影响。我们结合了三个不同的沿内分泌-免疫界面的应激指标来研究急性短期应激反应：总皮质醇浓度（Cort）、嗜中性粒细胞/淋巴细胞比值（H:L）和白细胞应对能力（LCC）。我们的研究提供了三个关键结果：i）皮质醇水平、LCC 和 H:L 之间没有关系，这证实了这三个参数代表应激反应中不同的生理终点；ii）所测量的参数对应激的反应动力学不同；iii）由于增加了一次采血或处理事件，在捕获后 30 分钟，生理应激水平没有差异。通过优化采样设计，我们的结果为鸟类物种的动物福利和应激生理学实验程序规划提供了启示。
