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Development of a whole-body cortisol extraction procedure for determination of stress in golden shiners, Notemigonus crysoleucas

Fish Physiology and Biochemistry volume 33, pages 189–193 (2007)Cite this article

Abstract

Baitfish such as golden shiners are subjected to stress during harvesting, grading, and transport. Their small size makes it difficult to measure the stress response with the biological indicator cortisol using conventional assay methods for plasma. This paper examines the development and validation of methods for whole-body cortisol extraction from individual baitfish. Three types of extracts were tested: (1) an ethyl ether unaltered extract (UA); (2) an extract reconstituted in phosphate buffered saline (PBS); (3) an extract that had been increased in volume by the addition of food-grade vegetable oil (VO). These extracts were evaluated using validation tests with radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA). The UA extract produced inadequate volumes of extract for multiple assays and could not be used for the determination of cortisol in a single fish. The PBS reconstitution method failed the precision recovery of serial dilutions (62.3%), linearity (R 2: 0.7864), and parallelism validation tests. The VO volume-boosting method passed all validation tests [intra-assay coefficent of variation (%CV): 16.3 for ELISA and 5.9 for RIA; inter-assay %CV: 10.3; spiked recovery: 102.0%; dilution recovery: 93.0%; linearity R 2: 0.9435; log of serial dilutions was parallel] and provided enough extract for multiple assays from an individual baitfish. Based on these results, we conclude that the VO volume-boosting method presents a means for determining cortisol from individual baitfish using either RIA or ELISA assays.

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Acknowledgements

The authors thank Kenneth Davis and Matthew McEntire of the United States Department of Agriculture’s Agriculture Research Service for their support in bringing this line of research to our attention.

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Authors and Affiliations

  1. Department of Aquaculture and Fisheries, University of Arkansas at Pine Bluff, 1200 North University Drive, mail slot 4912, Pine Bluff, AR, 71601, USA

    Todd David Sink, Sathyanand Kumaran & Rebecca T. Lochmann

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  1. Todd David Sink
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  2. Sathyanand Kumaran
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  3. Rebecca T. Lochmann
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Correspondence to Todd David Sink.

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Sink, T.D., Kumaran, S. & Lochmann, R.T. Development of a whole-body cortisol extraction procedure for determination of stress in golden shiners, Notemigonus crysoleucas . Fish Physiol Biochem 33, 189–193 (2007). https://doi.org/10.1007/s10695-007-9130-0

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  • DOI: https://doi.org/10.1007/s10695-007-9130-0

Keywords

  • Baitfish
  • Cortisol
  • Cortisol extraction
  • ELISA
  • Enzyme-linked immunosorbent assay
  • Golden shiners

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