e1826e9530a95a5133f01efac9ee1350 animals_a2025v15a762iENG.pdf 949decf9a97223647c13b78838730923544656f8 animals_a2025v15a762iENG.pdf 67983e8e0e5c4a0c0ec82946b963d356d8efb12d1c2c7bb966ae12213167837c animals_a2025v15a762iENG.pdf Title: A New Research Tool for Use in Sharks and Rays: Relevance of Reproductive Hormone Levels in the Skin of Small-Spotted Catshark (Scyliorhinus canicula) Subject: The present study marks a significant step forward in validating the use of skin biopsies for measuring reproductive steroid hormones in sharks. Given the conservation concerns surrounding many chondrichthyan species and the growing emphasis on adhering to the 3R principles in wildlife research, we should prioritize the acquisition of data through non-destructive or minimally invasive sampling techniques collected remotely. Dart skin biopsy samples can be collected remotely, without the need to chase, capture, and restrain the animal. Accordingly, the present study aimed to (i) develop a suitable extraction and analytical technique for reproductive hormone determination in shark skin biopsies and (ii) evaluate whether sex steroid hormones detected in shark skin differ between males and females and depending on males’ maturity. The results of assay validation demonstrated that the commercial enzyme immunoassay used can provide reliable measures of progesterone, 17-estradiol, and testosterone measured in shark skin extracts. After ensuring that reproductive hormones are accurately measured in this sample type, we compared hormone levels between males (mature and immature) and females to evaluate the variation in reproductive hormones relative to sex and males’ maturity stage. We detected significant differences in skin hormone levels between sexes, suggesting that this technique may be a promising alternative approach for obtaining relevant biological data in free-ranging sharks. By employing this novel technique, we expect to gain a deeper understanding of the reproductive processes of living chondrichthyans, which is essential for formulating effective science-based conservation and management strategies. Keywords: non-invasive sample; reproduction; testosterone; progesterone; 17-estradiol; chondrichthyans; conservation Author: Annaïs Carbajal, Isabel González Lobato, Clara Agustí, Marta Muñoz-Baquero, Paula Serres-Corral and Manel López-Béjar Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Thu Mar 6 16:12:34 2025 CET ModDate: Thu Mar 6 16:14:13 2025 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 11 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1203236 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- CAMXTK+EURM10 Type 1 Builtin yes yes yes 10 0 SSXPJJ+VnURWPalladioL Type 1 Custom yes yes yes 15 0 DZEQWQ+URWPalladioL-Roma Type 1 Custom yes yes yes 21 0 EAFJTB+URWPalladioL-Bold Type 1 Custom yes yes yes 27 0 RBDQNQ+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 LSAMXT+URWPalladioL-BoldItal Type 1 Custom yes yes yes 37 0 YFFKLH+CMSY10 Type 1 Builtin yes yes yes 66 0 CIDFont+F3 CID TrueType Identity-H yes no yes 82 0 CIDFont+F1 CID TrueType Identity-H yes no yes 90 0 CIDFont+F2 CID TrueType Identity-H yes no yes 98 0 CIDFont+F3 CID TrueType Identity-H yes no yes 110 0 CIDFont+F1 CID TrueType Identity-H yes no yes 118 0 CIDFont+F2 CID TrueType Identity-H yes no yes 126 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2025-04-04 02:42:28 CEST RepresentationInformation: animals_a2025v15a762iENG.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2025-04-03 20:22:31 CEST Size: 1203236 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 314 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: Animals and Sampling Destination: subsection.2.1 Item: Title: Hormone Extraction Destination: subsection.2.2 Item: Title: Hormone Analysis and Assay Validation Destination: subsection.2.3 Item: Title: Statistical Analysis Destination: subsection.2.4 Item: Title: Results Destination: section.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.6 Info: Title: A New Research Tool for Use in Sharks and Rays: Relevance of Reproductive Hormone Levels in the Skin of Small-Spotted Catshark (Scyliorhinus canicula) Author: Annaïs Carbajal, Isabel González Lobato, Clara Agustí, Marta Muñoz-Baquero, Paula Serres-Corral and Manel López-Béjar Subject: The present study marks a significant step forward in validating the use of skin biopsies for measuring reproductive steroid hormones in sharks. Given the conservation concerns surrounding many chondrichthyan species and the growing emphasis on adhering to the 3R principles in wildlife research, we should prioritize the acquisition of data through non-destructive or minimally invasive sampling techniques collected remotely. Dart skin biopsy samples can be collected remotely, without the need to chase, capture, and restrain the animal. Accordingly, the present study aimed to (i) develop a suitable extraction and analytical technique for reproductive hormone determination in shark skin biopsies and (ii) evaluate whether sex steroid hormones detected in shark skin differ between males and females and depending on males’ maturity. The results of assay validation demonstrated that the commercial enzyme immunoassay used can provide reliable measures of progesterone, 17-estradiol, and testosterone measured in shark skin extracts. After ensuring that reproductive hormones are accurately measured in this sample type, we compared hormone levels between males (mature and immature) and females to evaluate the variation in reproductive hormones relative to sex and males’ maturity stage. We detected significant differences in skin hormone levels between sexes, suggesting that this technique may be a promising alternative approach for obtaining relevant biological data in free-ranging sharks. By employing this novel technique, we expect to gain a deeper understanding of the reproductive processes of living chondrichthyans, which is essential for formulating effective science-based conservation and management strategies. Keywords: non-invasive sample; reproduction; testosterone; progesterone; 17-estradiol; chondrichthyans; conservation Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Thu Mar 06 16:12:34 CET 2025 ModDate: Thu Mar 06 16:14:13 CET 2025 ID: 0xde42fee2818cabda4f334446b981d8b0, 0xde42fee2818cabda4f334446b981d8b0 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: RBDQNQ+URWPalladioL-Ital FontSubset: true FirstChar: 44 LastChar: 122 FontDescriptor: FontName: RBDQNQ+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true ToUnicode: true Font: BaseFont: YFFKLH+CMSY10 FontSubset: true FirstChar: 0 LastChar: 48 FontDescriptor: FontName: YFFKLH+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true ToUnicode: true Font: BaseFont: LSAMXT+URWPalladioL-BoldItal FontSubset: true FirstChar: 75 LastChar: 121 FontDescriptor: FontName: LSAMXT+URWPalladioL-BoldItal Flags: Symbolic FontBBox: -170, 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