The role of water chemistry in zebrafish welfare and reproducibility of research studies

水化学在斑马鱼福利和研究再现性中的作用

• 批准号: NC/S001123/1
• 负责人: Rod Wilson
• 金额: $ 44.81万
• 依托单位: UNIVERSITY OF EXETER
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FS001123%2F1
• 关键词: role; water; chemistry; zebrafish; welfare

The present proposal will result in improvements relating to the Reduction and Refinement components of the 3Rs: REDUCTION: 1) Zebrafish research facilities will need to produce fewer stock animals to fulfil experimental numbers as the animals produced will be higher quality when water chemistry is optimised.2) Optimal water chemistry is also expected to improve the consistency of the quality of stock fish (i.e. less inter-individual variability). This will also reduce animal use overall because: a) less replication within studies required to achieve the same statistical power in detecting treatment differences, and b) the reproducibility between studies and different facilities will be improved. 3) Optimising water quality will also extend the timeframe of peak embryo production by adults, which in turn will reduce the frequency of producing new broodstock lines. This window currently varies from 4 to 9 months, so consistently working at the upper end of this range could halve the number of new broodstock lines required. REFINEMENT: This study will also minimise pain, suffering, distress or lasting harm associated with sub-optimal water quality. In particular elevated CO2 is known to impair brain function and the ability of fish to respond appropriately to sensory stimuli (olfactory, auditory and visual) that are part of their normal life and social interactions, and impairs learning and cognitive function, and can induce anxiety. The costs of coping with high CO2 and acid-base regulation also detract from energy spent on healthy growth, development, reproduction, immune function and disease resistance. The proposed study will provide evidence to optimise CO2 itself, as well as variables known to limit the ability of fish to achieve acid-base homeostasis associated everyday challenges of digestion/exercise/husbandry stresses and high CO2. The latter includes freshwater Na+ and Cl- levels which limit the rate of acid and base excretion by the gills, and calcium that directly regulates skin and gill permeability to all molecules. Water bicarbonate also determines the tolerance of fish to high CO2, and recovering their acid-base balance. The proposed experiments will therefore offer the opportunity to significantly reduce the detrimental impacts of water sub-optimal quality in zebrafish research facilities throughout the world. METRICS ON POTENTIAL 3Rs IMPACT: Until the research is completed it is difficult to precisely quantify the anticipated reduction of animals used and suffering experienced. However, zebrafish are the number one model species used in fish studies globally, with more than 3,250 institutes in 100 countries estimated to conduct zebrafish research. Zebrafish represent the 2nd most used species of any vertebrate in animal research, with >5 million fish used per annum. Therefore, even if only 1 % of all the global zebrafish research facilities adopted the guidelines we aim to produce, then at least 50,000 fish per year would benefit from improved welfare (which should include physiological homeostasis, growth, behaviour and immune function). Even if this 1 % figure only applied to the UK, then >4,000 zebrafish would still benefit per year. In reality the 1 % figure is likely to be a very conservative estimate of improved welfare for zebrafish globally. This will be due to fewer fish being required per study due to better consistency of production (i.e. lower inter-individual variability), and due to improved reproducibility between studies and facilities. In turn, this will translate into more robust and reliable conclusions regarding the outcomes of research (e.g. efficacy of drug treatments etc.). Regarding the "Refinement" component, if high quality guidelines are produced and disseminated effectively there is potential to reduce animal suffering on a truly global scale and improve the cumulative lifetime experience of millions of animals per year.

本提案将导致与3RS的减少和改进组成部分有关的改进：减少：1）斑马鱼研究设施将需要生产较少的库存动物以实现实验数字，因为当水化学的质量更高时，最佳水化学也将提高库存质量的质量，而不是库存的质量较小。这也将减少动物的总体使用，因为：a）在检测治疗差异时获得相同统计能力所需的研究中的复制减少，b）研究与不同设施之间的可重复性将得到改善。 3）优化水质还将延长成年人峰值产生的峰值胚胎的时间范围，这反过来将降低产生新的繁殖线的频率。目前，此窗口从4到9个月不等，因此在此范围的上端持续工作可能会使所需的新繁殖线的数量减半。 改进：这项研究还将最大程度地减少与亚最佳水质相关的疼痛，痛苦，痛苦或持久伤害。尤其是升高的二氧化碳是损害大脑功能的损害，以及鱼类对感觉刺激（嗅觉，听觉和视觉）适当反应的能力，这是其正常生活和社交互动的一部分，并损害学习和认知功能，并会引起焦虑。应对高二氧化碳和酸碱调节的成本也损害了在健康生长，发育，繁殖，免疫功能和抗病性上花费的能量。拟议的研究将提供证据以优化二氧化碳本身，以及已知的变量，以限制鱼类实现酸碱稳态相关的消化/运动/饲养应力和高二氧化碳的日常挑战的能力。后者包括淡水Na+和Cl-水平，这些水平限制了g的酸和碱排泄的速率，以及直接调节对所有分子的皮肤和g的渗透性的钙。水碳酸氢盐还决定了鱼对高二氧化碳的耐受性，并恢复了其酸碱平衡。因此，拟议的实验将提供机会，可显着减少全球斑马鱼研究设施中水质量的有害影响。潜在3RS影响的指标：在研究完成之前，很难精确地量化预期的动物所经历和苦难的预期减少。但是，斑马鱼是全球鱼类研究中使用的第一种模型物种，估计有100个国家的3,250多家机构进行斑马鱼研究。斑马鱼代表动物研究中任何脊椎动物中使用的第二种最常用的物种，每年使用> 500万只鱼。因此，即使只有所有全球斑马鱼研究设施中只有1％采用了我们旨在生产的准则，那么每年至少有50,000条鱼将受益于改善福利（包括生理体内平衡，成长，行为和免疫功能）。即使这个1％的数字仅适用于英国，那么> 4,000斑马鱼仍将每年受益。实际上，1％的数字可能是全球斑马鱼福利改善的非常保守的估计。这将是由于每项研究所需的鱼类所需的较少，因为生产的一致性更好（即个体间的变异性降低），并且由于研究和设施之间的可重复性提高了。反过来，这将转化为有关研究结果（例如，药物治疗等）的更强大和可靠的结论。关于“改进”组成部分，如果生产高质量的指南并有效地传播，则有可能在真正的全球范围内减少动物苦难并改善每年数百万只动物的累积终身经验。