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Methods to collect, preserve, and prepare elasmobranch tissues for stable isotope analysis

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Abstract

Stable isotope analysis has the potential to expand our understanding of elasmobranch ecology. However, elasmobranchs share unique traits (i.e., retention of urea, lack of adipose tissue, cartilaginous skeletons) that require modified preparation techniques. Alternative tissue collection and preservation methods would allow sampling from ichthyology collections and at remote locations. We compared different collection, preservation, and preparation techniques to identify treatments that yielded robust isotopic data. Blood components collected in tubes coated with lithium heparin (an anti-coagulant) were not isotopically distinct from blood collected in no-additive tubes. Compared to frozen muscle, ethanol-treated muscle had altered δ13C values, but similar δ15N values. Finally, we removed lipids and urea with petroleum ether and deionized water, respectively. Although untreated and treated muscle had similar amino acid compositions, treated muscle preferentially lost 14N and had greater C:N ratios. These results indicate that urea affects isotope ratios and that water treatment removes urea without altering muscle protein composition. Although not exhaustive, our study begins to address the need for elasmobranch-specific methods.

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Acknowledgements

We thank L. Kroll, S. Perry, S. Rumbolt, A. Sjostrom, A. Thell, and C. Spencer for aquarium and sampling assistance, F. Batista for amino acid analysis. We also thank L. Roland, D. Shizuka, and two anonymous reviews for their comments and critical reviews. D. Casper, D.V.M. at Long Marine Lab (UCSC) facilitated the successful rearing of leopard sharks in captivity. All sampling methods for leopard sharks followed the Guide to the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee at UCSC. The skates were caught by the Pacific Shark Research Center at MLML during regular survey trawls. Financial support provided by the Institute of Geophysics and Planetary Physics at UCSC and the Myers Ocean Conservation Grant.

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  1. Sora L. Kim

    Present address: Department of Geology and Geophysics, University of Wyoming, 1000 University Ave. #3006, Laramie, WY, 82071, USA

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  1. Department of Earth and Planetary Sciences, University of California, Santa Cruz, 1156 High St, Santa Cruz, CA, 95064, USA

    Sora L. Kim & Paul L. Koch

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  1. Sora L. Kim
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Correspondence to Sora L. Kim.

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ESM 1

The mole percents of individual AAs in plasma, RBC, and muscle with treatment are listed. (DOC 17 kb)

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Kim, S.L., Koch, P.L. Methods to collect, preserve, and prepare elasmobranch tissues for stable isotope analysis. Environ Biol Fish 95, 53–63 (2012). https://doi.org/10.1007/s10641-011-9860-9

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  • DOI: https://doi.org/10.1007/s10641-011-9860-9

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