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| Content Provider | Springer Nature Link |
|---|---|
| Author | Sink, Todd David Kumaran, Sathyanand Lochmann, Rebecca T. |
| Copyright Year | 2007 |
| 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. |
| Starting Page | 189 |
| Ending Page | 193 |
| Page Count | 5 |
| File Format | |
| ISSN | 09201742 |
| Journal | Fish Physiology and Biochemistry |
| Volume Number | 33 |
| Issue Number | 3 |
| e-ISSN | 15735168 |
| Language | English |
| Publisher | Kluwer Academic Publishers |
| Publisher Date | 2007-02-22 |
| Publisher Place | Dordrecht |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Baitfish Cortisol Cortisol extraction ELISA Enzyme-linked immunosorbent assay Golden shiners Zoology Animal Biochemistry Animal Anatomy / Morphology / Histology Animal Physiology Hydrobiology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Physiology Medicine Biochemistry Aquatic Science |
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