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Validating the underpinnings of water corticosterone measurement for aquatic amphibians

Smith, Tessa E.
; 
Holmes, Andrew M
; 
Emmans, Christopher J
; 
Coleman, Robert
; 
Hosie, Charlotte A.
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Affiliation
University of Chester
EPub Date
Publication Date
2025-01-09
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Biological Sciences
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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 Xenopus, including the influence of circadian rhythm, sex and snout-vent length on waterborne corticosterone levels, since fundamental knowledge of a species’ biology is essential for designing robust experiments and in the interpretation of the results. 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.
Citation
Smith, T. E., Holmes, A. M., Emmans, C. J., Coleman, R., & Hosie, C. A. (2025). Validating the underpinnings of water corticosterone measurement for aquatic amphibians. F1000Research, 14, 53. https://doi.org/10.12688/f1000research.157055.1
Publisher
Taylor and Francis
Journal
F1000Research
Research Unit
URI
http://hdl.handle.net/10034/629216
DOI
10.12688/f1000research.157055.1
PubMed ID
PubMed Central ID
Type
Article
Language
en
Description
Series/Report no.
ISSN
2046-1402
EISSN
2046-1402
ISBN
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National Centre for the Replacement Refinement and Reduction of Animals in Research
Additional Links
https://f1000research.com/articles/14-53/v1