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RationaleIt is imperative to understand how chemical preservation alters tissue isotopic compositions before using historical samples in ecological studies. Specifically, although compound‐specific isotope analysis of amino acids (CSIA‐AA) is becoming a widely used tool, there is little information on how preservation techniques affect amino acidδ15N values. MethodsWe evaluated the effects of chemical preservatives on bulk tissueδ13C andδ15N and amino acidδ15N values, measured by gas chromatography/isotope ratio mass spectrometry (GC/IRMS), of (a) tuna (Thunnus albacares) and squid (Dosidicus gigas) muscle tissues that were fixed in formaldehyde and stored in ethanol for 2 years and (b) two copepod species,Calanus pacificusandEucalanus californicus, which were preserved in formaldehyde for 24–25 years. ResultsTissues in formaldehyde‐ethanol had higher bulkδ15N values (+1.4,D. gigas; +1.6‰,T. albacares), higherδ13C values forD. gigas(+0.5‰), and lowerδ13C values forT. albacares(−0.8‰) than frozen samples. The bulkδ15N values from copepods were not different those from frozen samples, although theδ13C values from both species were lower (−1.0‰ forE. californicusand −2.2‰ forC. pacificus) than those from frozen samples. The mean amino acidδ15N values from chemically preserved tissues were largely within 1‰ of those of frozen tissues, but the phenylalanineδ15N values were altered to a larger extent (range: 0.5–4.5‰). ConclusionsThe effects of preservation on bulkδ13C values were variable, where the direction and magnitude of change varied among taxa. The changes in bulkδ15N values associated with chemical preservation were mostly minimal, suggesting that storage in formaldehyde or ethanol will not affect the interpretation ofδ15N values used in ecological studies. The preservation effects on amino acidδ15N values were also mostly minimal, mirroring bulkδ15N trends, which is promising for future CSIA‐AA studies of archived specimens. However, there were substantial differences in phenylalanine and valineδ15N values, which we speculate resulted from interference in the chromatographic resolution of unknown compounds rather than alteration of tissue isotopic composition due to chemical preservation.
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The influence of biopsy site and pregnancy on stable isotope ratios in humpback whale skin
Abstract RationaleStable isotope analysis (SIA) of free‐swimming mysticetes using biopsies is often limited in sample size and uses only one sample per individual, failing to capture both intra‐individual variability and the influence of demographic and physiological factors on isotope ratios. MethodsWe applied SIA of δ13C and δ15N to humpback whale (Megaptera novaeangliae) biopsies taken during the foraging season along the western Antarctic Peninsula to quantify intra‐individual variation from repeatedly sampled individuals, as well as to determine the effect of biopsy collection site, sex, and pregnancy on isotope ratios. ResultsThere was substantial variability in δ13C from multiple biopsies taken from the same individuals, though δ15N was much more consistent. Side of the body (left versus right) and biopsy location (dorsal, anterior, ventral, and posterior) did marginally affect the isotopic composition of δ15N but not δ13C. Pregnancy had a significant effect on both δ13C and δ15N, where pregnant females were depleted in both when compared to non‐pregnant females and males. ConclusionsThese results indicate that isotopic signatures are influenced by multiple endogenous and exogenous factors and emphasize value in accounting for intra‐individual variability and pregnancy status within a sampled population. Placed within an ecological context, the endogenous variability in δ13C observed here may be informative for future isotopic analyses.
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- Award ID(s):
- 1927742
- PAR ID:
- 10499437
- Publisher / Repository:
- Wiley Blackwell (John Wiley & Sons)
- Date Published:
- Journal Name:
- Rapid Communications in Mass Spectrometry
- Volume:
- 38
- Issue:
- 11
- ISSN:
- 0951-4198
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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