016e34955d91975f43e9759421a4eaba pmc_11083227.pdf 68fb14aa27ad5d482349ec8eb7c2dbe8e95e65d4 pmc_11083227.pdf 79e1aee74a8c35a85d314dae54a583edcb991cb8b0fb5c691c9b383f8af47afb pmc_11083227.pdf Title: Validating a Non-Invasive Method for Assessing Cortisol Concentrations in Scraped Epidermal Skin from Common Bottlenose Dolphins and Belugas Subject: Society is showing a growing concern about the welfare of cetaceans in captivity as well as cetaceans in the wild threatened by anthropogenic disturbances. The study of the physiological stress response is increasingly being used to address cetacean conservation and welfare issues. Within it, a newly described technique of extracting cortisol from epidermal desquamation may serve as a non-invasive, more integrated measure of a cetacean’s stress response and welfare. However, confounding factors are common when measuring glucocorticoid hormones. In this study, we validated a steroid hormone extraction protocol and the use of a commercial enzyme immunoassay (EIA) test to measure cortisol concentrations in common bottlenose dolphin (Tursiops truncatus) and beluga (Delphinapterus leucas) epidermal samples. Moreover, we examined the effect of sample mass and body location on cortisol concentrations. Validation tests (i.e., assay specificity, accuracy, precision, and sensitivity) suggested that the method was suitable for the quantification of cortisol concentrations. Cortisol was extracted from small samples (0.01 g), but the amount of cortisol detected and the variability between duplicate extractions increased as the sample mass decreased. In common bottlenose dolphins, epidermal skin cortisol concentrations did not vary significantly across body locations while there was a significant effect of the individual. Overall, we present a contribution towards advancing and standardizing epidermis hormone assessments in cetaceans. Keywords: cortisol; skin; stratum corneum; keratin; steroid hormones; cetacean; stress;conservation biology Author: Clara Agustí, Xavier Manteca, Daniel García-Párraga and Oriol Tallo-Parra Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Fri May 3 11:34:04 2024 CEST ModDate: Fri May 3 11:39:54 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2128975 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SSNIUI+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 XFKEXT+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 YEISVM+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 DENMTT+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 NBMIUS+VnURWPalladioL Type 1 Custom yes yes yes 32 0 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02:35:40 CEST Size: 2128975 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 721 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Experimental Design Destination: subsection.2.1 Item: Title: Epidermal Sampling Destination: subsection.2.2 Item: Title: Sample Preparation and Storage Destination: subsection.2.3 Item: Title: Epidermis Pool, Effect of Sample Mass, and Sample Processing Error Destination: subsection.2.4 Item: Title: Effect of Body Location Destination: subsection.2.5 Item: Title: Hormone Extraction Destination: subsection.2.6 Item: Title: Hormone Detection and Assay Validation Destination: subsection.2.7 Item: Title: Statistical Analyses Destination: subsection.2.8 Item: Title: Results Destination: section.3 Children: Item: Title: Validation of the Epidermis Collection Methodology and the EIA Destination: subsection.3.1 Item: Title: Effect of Sample Mass Destination: subsection.3.2 Item: Title: Effect of Individual, Body Location, and Sex in Common Bottlenose Dolphins Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Children: Item: Title: Validation of the Epidermis Collection Methodology and the EIA Destination: subsection.4.1 Item: Title: Effect of Sample Mass Destination: subsection.4.2 Item: Title: Effect of Individual, Body Location, and Sex Destination: subsection.4.3 Item: Title: Cetacean Epidermis as a Storage Medium of Steroid Hormones Destination: subsection.4.4 Item: Title: Implications of the Study and Future Research Destination: subsection.4.5 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.6 Info: Title: Validating a Non-Invasive Method for Assessing Cortisol Concentrations in Scraped Epidermal Skin from Common Bottlenose Dolphins and Belugas Author: Clara Agustí, Xavier Manteca, Daniel García-Párraga and Oriol Tallo-Parra Subject: Society is showing a growing concern about the welfare of cetaceans in captivity as well as cetaceans in the wild threatened by anthropogenic disturbances. The study of the physiological stress response is increasingly being used to address cetacean conservation and welfare issues. Within it, a newly described technique of extracting cortisol from epidermal desquamation may serve as a non-invasive, more integrated measure of a cetacean’s stress response and welfare. However, confounding factors are common when measuring glucocorticoid hormones. In this study, we validated a steroid hormone extraction protocol and the use of a commercial enzyme immunoassay (EIA) test to measure cortisol concentrations in common bottlenose dolphin (Tursiops truncatus) and beluga (Delphinapterus leucas) epidermal samples. Moreover, we examined the effect of sample mass and body location on cortisol concentrations. Validation tests (i.e., assay specificity, accuracy, precision, and sensitivity) suggested that the method was suitable for the quantification of cortisol concentrations. Cortisol was extracted from small samples (0.01 g), but the amount of cortisol detected and the variability between duplicate extractions increased as the sample mass decreased. In common bottlenose dolphins, epidermal skin cortisol concentrations did not vary significantly across body locations while there was a significant effect of the individual. Overall, we present a contribution towards advancing and standardizing epidermis hormone assessments in cetaceans. 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Annotation: Subtype: Link Rect: 506, 189, 542, 200 Annotation: Subtype: Link Rect: 176, 167, 213, 177 Annotation: Subtype: Link Rect: 219, 167, 255, 177 Annotation: Subtype: Link Rect: 218, 143, 255, 154 Annotation: Subtype: Link Rect: 261, 143, 297, 154 Annotation: Subtype: Link Rect: 447, 120, 484, 131 Annotation: Subtype: Link Rect: 491, 120, 527, 131 Annotation: Subtype: Link Rect: 261, 97, 298, 108 Page: Sequence: 18 Annotations: Annotation: Subtype: Link Rect: 104, 730, 141, 741 Annotation: Subtype: Link Rect: 147, 730, 183, 741 Annotation: Subtype: Link Rect: 95, 696, 133, 706 Annotation: Subtype: Link Rect: 139, 696, 175, 706 Annotation: Subtype: Link Rect: 437, 683, 474, 694 Annotation: Subtype: Link Rect: 59, 661, 96, 671 Annotation: Subtype: Link Rect: 153, 638, 191, 648 Annotation: Subtype: Link Rect: 440, 614, 478, 625 Annotation: Subtype: Link Rect: 484, 614, 520, 625 Annotation: Subtype: Link Rect: 459, 591, 497, 602 Annotation: Subtype: Link Rect: 503, 591, 539, 602 Annotation: Subtype: Link Rect: 223, 557, 260, 568 Annotation: Subtype: Link Rect: 267, 557, 303, 568 Annotation: Subtype: Link Rect: 471, 534, 509, 545 Annotation: Subtype: Link Rect: 515, 534, 551, 545 Annotation: Subtype: Link Rect: 302, 499, 340, 510 Annotation: Subtype: Link Rect: 346, 499, 382, 510 Annotation: Subtype: Link Rect: 146, 476, 184, 487 Annotation: Subtype: Link Rect: 190, 476, 226, 487 Annotation: Subtype: Link Rect: 486, 465, 524, 476 Annotation: Subtype: Link Rect: 91, 443, 128, 453 Annotation: Subtype: Link Rect: 134, 443, 170, 453 Annotation: Subtype: Link Rect: 153, 420, 191, 430 Annotation: Subtype: Link Rect: 254, 397, 291, 407 Annotation: Subtype: Link Rect: 113, 362, 150, 372 Annotation: Subtype: Link Rect: 227, 339, 265, 349 Annotation: Subtype: Link Rect: 398, 315, 436, 326 Annotation: Subtype: Link Rect: 458, 292, 496, 303 Checksum: f147a7cd Type: CRC32 Checksum: 016e34955d91975f43e9759421a4eaba Type: MD5 Checksum: 68fb14aa27ad5d482349ec8eb7c2dbe8e95e65d4 Type: SHA-1 Checksum: 79e1aee74a8c35a85d314dae54a583edcb991cb8b0fb5c691c9b383f8af47afb Type: SHA-256