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Predictable weathering of puparial hydrocarbons of necrophagous flies for determining the postmortem interval: a field experiment using Chrysomya rufifacies


Preadult development of necrophagous flies is commonly recognized as an accurate method for estimating the minimum postmortem interval (PMImin). However, once the PMImin exceeds the duration of preadult development, the method is less accurate. Recently, fly puparial hydrocarbons were found to significantly change with weathering time in the field, indicating their potential use for PMImin estimates. However, additional studies are required to demonstrate how the weathering varies among species. In this study, the puparia of Chrysomya rufifacies were placed in the field to experience natural weathering to characterize hydrocarbon composition change over time. We found that weathering of the puparial hydrocarbons was regular and highly predictable in the field. For most of the hydrocarbons, the abundance decreased significantly and could be modeled using a modified exponent function. In addition, the weathering rate was significantly correlated with the hydrocarbon classes. The weathering rate of 2-methyl alkanes was significantly lower than that of alkenes and internal methyl alkanes, and alkenes were higher than the other two classes. For mono-methyl alkanes, the rate was significantly and positively associated with carbon chain length and branch position. These results indicate that puparial hydrocarbon weathering is highly predictable and can be used for estimating long-term PMImin.

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  1. Chisum WJ, Turvey BE (2011) Chapter 8 - Methods of crime reconstruction. In: Crime Reconstruction, 2nd edn. Academic Press, San Diego, pp 179–209

  2. DuPre DMP (2013) Chapter 2 - Basic investigation protocols. In: Homicide Investigation Field Guide. Academic Press, San Diego, pp 5–24

  3. Henssge C, Madea B (2007) Estimation of the time since death. Forensic Sci Int 165:182–184

    Article  PubMed  Google Scholar 

  4. Henssge C, Madea B (2004) Estimation of the time since death in the early post-mortem period. Forensic Sci Int 144:167–175

    CAS  Article  PubMed  Google Scholar 

  5. Tomberlin JK, Benbow ME (eds) (2015) Forensic Entomology: International Dimensions and Frontiers (Contemporary Topics in Entomology). CRC Press, Boca Raton, pp 422

  6. Anderson GS (2010) Chapter 5. Factors that influence insect succession on carrion. In: Byrd JH, Castner JL (eds) Forensic Entomology: the Utility of Arthropods in Legal Investigations, 2nd edn. CRC Press, Boca Raton, pp 201–250

  7. Amendt J, Krettek R, Zehner R (2004) Forensic entomology. Naturwissenschaften 91:51–65

    CAS  Article  PubMed  Google Scholar 

  8. Catts EP, Goff ML (1992) Forensic entomology in criminal investigations. Annu Rev Entomol 37:253–272

    CAS  Article  PubMed  Google Scholar 

  9. Tomberlin JK, Mohr R, Benbow ME et al (2011) A roadmap for bridging basic and applied research in forensic entomology. Annu Rev Entomol 56:401–421

    CAS  Article  PubMed  Google Scholar 

  10. Lewis AJ, Benbow ME (2011) When entomological evidence crawls away: Phormia regina en masse larval dispersal. J Med Entomol 48:1112–1119

    CAS  Article  PubMed  Google Scholar 

  11. Zhu GH, Xu XH, Yu XJ et al (2007) Puparial case hydrocarbons of Chrysomya megacephala as an indicator of the postmortem interval. Forensic Sci Int 169:1–5

    CAS  Article  PubMed  Google Scholar 

  12. Zhu GH, Yu XJ, Xie LX et al (2013) Time of death revealed by hydrocarbons of empty puparia of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae): a field experiment. PLoS One 8:e73043

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. Archer MS, Elgar MA (2003) Yearly activity patterns in southern Victoria (Australia) of seasonally active carrion insects. Forensic Sci Int 132:173–176

    CAS  Article  PubMed  Google Scholar 

  14. Braga MV, Pinto ZT, Queiroz MMD et al (2013) Cuticular hydrocarbons as a tool for the identification of insect species: puparial cases from Sarcophagidae. Acta Trop 128:479–485

    CAS  Article  PubMed  Google Scholar 

  15. Goodrich BS (1970) Cuticular lipids of adults and puparia of the Australian sheep blowfly Lucilia cuprina (Wied.). J Lipid Res 11:1–6

    CAS  PubMed  Google Scholar 

  16. Ye GY, Li K, Zhu JY et al (2007) Cuticular hydrocarbon composition in pupal exuviae for taxonomic differentiation of six necrophagous flies. J Med Entomol 44:450–456

    CAS  Article  PubMed  Google Scholar 

  17. Musah RA, Espinoza EO, Cody RB et al (2015) A high throughput ambient mass spectrometric approach to species identification and classification from chemical fingerprint signatures. Sci Rep 5:11520

    Article  PubMed  PubMed Central  Google Scholar 

  18. Frere B, Suchaud F, Bernier G et al (2014) GC-MS analysis of cuticular lipids in recent and older scavenger insect puparia. An approach to estimate the postmortem interval (PMI). Anal Bioanal Chem 406:1081–1088

    CAS  Article  PubMed  Google Scholar 

  19. Drijfhout FP (2010) Chapter 10. Cuticular hydrocarbons: a new tool in forensic entomology? In: Amendt J, Goff ML, Campobasso CP, Grassberger M (eds) Current Concepts in Forensic Entomology. Springer Science + Business Media, New York, pp 179–203

  20. Bacosa HP, Liu ZF, Erdner DL (2015) Natural sunlight shapes crude oil-degrading bacterial communities in northern Gulf of Mexico waters. Front Microbiol 6:1325

    Article  PubMed  PubMed Central  Google Scholar 

  21. Sharma P, Schiewer S (2016) Assessment of crude oil biodegradation in arctic seashore sediments: effects of temperature, salinity, and crude oil concentration. Environ Sci Pollut R 23:14881–14888

    CAS  Article  Google Scholar 

  22. Blomquist GJ, Nelson DR, Renobales MD (1987) Chemistry, biochemistry, and physiology of insect cuticular lipids. Arch Insect Biochem Physiol 6:227–265

    CAS  Article  Google Scholar 

  23. Gibbs AG (1998) Water-proofing properties of cuticular lipids. Am Zool 38:471–482

    CAS  Article  Google Scholar 

  24. Lockey KH (1988) Lipids of the insect cuticle: origin, composition and function. Comp Biochem Physiol 89B:595–645

    CAS  Google Scholar 

  25. Lockey KH (1991) Insect hydrocarbon classes: implications for chemotaxonomy. Insect Biochem 21:91–97

    CAS  Article  Google Scholar 

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This study was funded by grants from the National Natural Science Foundation of China ( (0700967), Natural Science Foundation of Guangdong, China ( (06301086), and China Postdoctoral Science Foundation ( (2004035616). The time for MEB was supported by Michigan State University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Correspondence to Guang-Hui Zhu.

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Zhu, GH., Jia, ZJ., Yu, XJ. et al. Predictable weathering of puparial hydrocarbons of necrophagous flies for determining the postmortem interval: a field experiment using Chrysomya rufifacies . Int J Legal Med 131, 885–894 (2017).

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  • Chrysomya rufifacies
  • Puparial hydrocarbons
  • Weathering time
  • Postmortem interval
  • Forensic entomology