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Analysis of the 13C isotope ratios of amino acids in the larvae, pupae and adult stages of Calliphora vicina blow flies and their carrion food sources

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Abstract

Adult blow flies are one of the first necrophagous insects to colonize fresh carcasses. The eggs they lay hatch into larvae, which then feed on the decomposing body. Like all organisms, blow flies “are what they eat,” meaning that the isotopic composition of their body tissues reflects their diet. This manuscript combines ecology with a forensic application by using isotope ratio mass spectrometry (IRMS) to understand the relationship between the δ13C of amino acids in different carrion sources and the blow fly that feed on them. We also measure the amino acid-level fractionation that occurs at each major life stage of the blow flies. Adult blow flies from a commercial strain of Calliphora vicina (Robineau-Desvoidy) (Diptera: Calliphoridae) oviposited on raw pork muscle, beef muscle, or chicken liver. Larvae, pupae, and adult blow flies from each carrion were selected for amino acid compound-specific isotope analysis. Canonical discriminant analysis showed that flies were correctly classified to specific carrion types in 100% (original rules) and 96.8% (leave-one-out cross-validation [LOOCV]) of cases. Regarding life stages, we obtained 100% and 71% of correct classification in original rules and LOOCV, respectively. The isotope ratios of most of the essential amino acids did not significantly change between life stages (at 95% CI). However, some non-essential amino acids (Ala, Ser, and Glu) and some conditionally essential amino acids (Gly and Pro) were isotopically depleted in the adult stage. Except for the essential amino acids, the amino acids in larvae and pupae were enriched in 13C, and adult blow flies were depleted in 13C relative to the carrion on which they fed. These results make it possible to exclude potential sources of carrion as larval food. Amino acid-specific IRMS could help inform entomologists whether a fly has just arrived from another location to feed on a corpse or has emerged from a pupa whose feedstock was the corpse. Such insight could enhance the significance of blow flies for post-mortem interval determinations. The analytical ability to link organisms from one trophic level to another through the use of compound-specific isotope analysis of amino acids could have wide-reaching consequences in a variety of disciplines.

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Acknowledgements

We would like to thank Dr. Rita Rio, from the Department of Biology at West Virginia University, for her valuable insights about amino acids and their metabolism in insects. We also appreciate the valuable feedback form the external reviewers.

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Authors and Affiliations

  1. Department of Biology, West Virginia University, Morgantown, WV, 26506-6121, USA

    Mayara P. V. Matos

  2. C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, WV, 26506-6121, USA

    Kateryna I. Konstantynova & Glen P. Jackson

  3. Department of Forensic and Investigative Science, West Virginia University, Morgantown, WV, 26506-6121, USA

    Rachel M. Mohr & Glen P. Jackson

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  1. Mayara P. V. Matos
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  2. Kateryna I. Konstantynova
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  4. Glen P. Jackson
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Correspondence to Glen P. Jackson.

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No human subjects were used in this research, and no live animals were used. Animal flesh (meat) was purchased from grocery stores.

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The authors declare that they have no conflict of interest.

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Matos, M.P.V., Konstantynova, K.I., Mohr, R.M. et al. Analysis of the 13C isotope ratios of amino acids in the larvae, pupae and adult stages of Calliphora vicina blow flies and their carrion food sources. Anal Bioanal Chem 410, 7943–7954 (2018). https://doi.org/10.1007/s00216-018-1416-9

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