ArticleF1000Research2025
Validating the underpinnings of water corticosterone measurement for aquatic amphibians.
Article in F1000Research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Authors and funding
5 authors.
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Abstract
Background: Good animal welfare is important ethically but also to ensure animals provide valid scientific models. Despite thousands of amphibians in research laboratories there is minimal quantitative evidence pertaining to their management and welfare. This study validated methods to non-invasively measure corticosterone, the amphibian 'stress' hormone, from tank water to provide a robust and reliable welfare assessment tool. Methods: We report experiments (A) that evaluate parameters linked to the performance of our biochemical extraction methods for waterborne corticosterone and, importantly, associated sampling procedures. We evaluate appropriate sampling water type, sampling vessel, filtration methods, potential degradation of waterborne corticosterone over time and the impact of sampling procedures on animal corticosterone levels. We wanted to determine sampling parameters that yielded the least background corticosterone and had minimum negative impact on the animals. The second series of experiments (B) evaluated parameters linked to the biology of Results: We propose collecting corticosterone samples in deionised water in either plastic or glass containers. The filtering process does not impact the amount of corticosterone measured in the water sample. Levels of corticosterone collected in the water change over a 48-hr period so we advocate standardising time from hormone collection to storage at - 20 °C. Repeated transfer of frogs to sampling containers does not increase corticosterone, suggesting our methods are not cumulatively stressful. Corticosterone levels were not impacted by circadian phase, sex or snout-vent length. Conclusion: We have developed and validated robust methods to quantify waterborne corticosterone. We hope they provide a template for researchers wishing to develop methods to measure waterborne corticosterone in aquatic amphibians.
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