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Validation of a DNA-based method for identifying Chrysomyinae (Diptera: Calliphoridae) used in a death investigation

  • Jeffrey D. Wells
  • Diana W. Williams
Original Article

Abstract

Many authors have proposed DNA-based methods for identifying an insect specimen associated with human remains. However, almost no attempt has been made to validate these methods using additional observations. We tested a protocol for identifying insects in the blow fly subfamily Chrysomyinae (Diptera: Calliphoridae) often found to be associated with a human corpse in Canada or the USA. This method uses phylogenetic analysis of DNA sequence from a short segment of the mitochondrial gene for cytochrome oxidase one (COI). Test chrysomyine COI sequences were obtained from 245 newly sequenced specimens and 51 specimens from the published literature. Published sequences from representatives of nonchrysomyine genera were also included to check for the possibility of a false positive identification. All of the chrysomyine test haplotypes were correctly identified with strong statistical support, and there were no false positives. This method appears to be an accurate and robust technique for identifying chrysomyine species from a death investigation in this geographic region. The far northern species Protophormia atriceps was not evaluated; therefore, caution is required in applying this method at very high latitudes in North America.

Keywords

Forensic entomology Mitochondrial DNA Cytochrome oxidase Phylogeny Species determination 

Notes

Many friends and colleagues either provided fly specimens or hosted JDW during a collecting trip. These included Whitney Banks (West Virginia U.), Michael Caterino (Santa Barbara Museum of Natural History), members of the Contra Costa, CA, Sheriff’s Department, Patrick Erwin (U. Alabama at Birmingham), Emmanuel Gaudry (French Gendarmerie), M. Lee Goff (Chaminade U.), Jack Hayes (U. Texas at San Antonio), Hiromu Kurahashi (Japanese National Institute of Infectious Disease), Wayne Lord (Federal Bureau of Investigation), Paul Richards (U. Miami), Daniel Rubinoff (U. Hawaii), Nikolaos Schizas (U. Puerto Rico Mayaguez), and Marta Wollf (U. Antioquia). This work was supported in part by the US National Institute of Justice award 99-IJ-0034. The views expressed here are not necessarily those of the US Department of Justice.

References

  1. 1.
    Ash N, Greenberg B (1975) Developmental temperature responses of the sibling species Phaenicia sericata and Phaenicia pallescens. Ann Entomol Soc Am 68:197–200Google Scholar
  2. 2.
    Baumgartner DL, Greenberg B (1984) The genus Chrysomya (Diptera: Calliphoridae) in the New World. J Med Entomol 21:105–113Google Scholar
  3. 3.
    Benecke M, Wells JD (2001) DNA techniques for forensic entomology. In: Byrd JH, Castner JL (eds) Forensic entomology: utility of arthropods in legal investigations. CRC Press, Boca Raton, pp 341–352Google Scholar
  4. 4.
    Bernasconi MV, Valsangiacomo C, Piffaretti JC, Ward PI (2000) Phylogenetic relationships among muscoidea (Diptera: calyptratae) based on mitochondrial DNA sequences. Insect Mol Biol 9:67–74CrossRefPubMedGoogle Scholar
  5. 5.
    Birstein VJ, Doukakis P, DeSalle R (2000) Polyphyly of mtDNA lineages in the Russian sturgeon, Acipenser gueldenstaedtii: forensic and evolutionary implications. Conserv Genet 1:81–88CrossRefGoogle Scholar
  6. 6.
    Bottero MT, Civera T, Anastasio A, Turi RM, Rosati S (2002) Identification of cow’s milk in “buffalo” cheese by duplex polymerase chain reaction. J Food Prot 65:362–366PubMedGoogle Scholar
  7. 7.
    Caterino MS, Cho S, Sperling FAH (2000) The current state of insect molecular systematics: a thriving Tower of Babel. Annu Rev Entomol 45:1–54PubMedCrossRefGoogle Scholar
  8. 8.
    Chen WY, Hung TH, Shiao SF (2004) Molecular identification of forensically important blow fly species (Diptera: Calliphoridae) in Taiwan. J Med Entomol 41:47–57PubMedCrossRefGoogle Scholar
  9. 9.
    Cronin ME, Palmisciano DA, Vyse ER, Cameron DG (1991) Mitochondrial DNA in wildlife forensic science: species identification of tissues. Wildl Soc Bull 19:94–105Google Scholar
  10. 10.
    Grassberger M, Friedrich E, Reiter C (2003) The blowfly Chrysomya albiceps (Wiedemann) (Diptera: Calliphoridae) as a new forensic indicator in central Europe. Int J Legal Med 117:75–81PubMedGoogle Scholar
  11. 11.
    Guimaraes J, do Prado A, Linhares AX (1978) Three newly introduced blowfly species in southern Brazil (Diptera: Calliphoridae). Rev Bras Entomol 22:53–60Google Scholar
  12. 12.
    Hall DG (1948) Blowflies of North America. Thomas Say Foundation, LafayetteGoogle Scholar
  13. 13.
    Hall DG (1965) Calliphoridae. In: Stone A, Sabrosky CW, Wirth WW, Foote RH, Coulson JR (eds) A catalog of the Diptera of America north of Mexico. Smithsonian Institution Press, Washington, pp 922–932Google Scholar
  14. 14.
    Harvey ML, Dadour IR, Gaudieri S (2003a) Mitochondrial DNA cytochrome oxidase I gene: potential for distinction between immature stages of some forensically important fly species (Diptera) in western Australia. Forensic Sci Int 131:134–139CrossRefPubMedGoogle Scholar
  15. 15.
    Harvey ML, Mansell MW, Villet MH, Dadour IR (2003b) Molecular identification of some forensically important blowflies of southern Africa and Australia. Med Vet Entomol 17:363–369CrossRefPubMedGoogle Scholar
  16. 16.
    Jiron LF (1979) Sobre moscas califoridas de Costa Rica (Diptera: Cyclorrhapha). Brenesia 16:221–222Google Scholar
  17. 17.
    Junqueira AC, Lessinger AC, Torres TT, da Silva FR, Vettore AL, Arruda P, Azeredo Espin AM (2004) The mitochondrial genome of the blowfly Chrysomya chloropyga (Diptera: Calliphoridae). Gene 339:7–15CrossRefPubMedGoogle Scholar
  18. 18.
    Kamal AS (1958) Comparative study of thirteen species of sarcosaprophagous Calliphoridae and Sarcophagidae (Diptera). 1. Bionomics. Ann Entomol Soc Am 51:261–270Google Scholar
  19. 19.
    Lenstra JA (2003) DNA methods for identifying plant and animals species in foods. In: Lees M (ed) Food authenticity and traceability. CRC Press, Boca Raton, pp 34–53Google Scholar
  20. 20.
    Malgorn Y, Coquoz R (1999) DNA typing for identification of some species of Calliphoridae. An interest in forensic entomology. Forensic Sci Int 102:111–119CrossRefPubMedGoogle Scholar
  21. 21.
    Marko PB, Lee SC, Rice AM, Gramling JM, Fitxhenry TM, McAllister JS, Harper GR, Moran A (2004) Mislabelling of a depleted reef fish. Nature 430:309–310CrossRefPubMedGoogle Scholar
  22. 22.
    Moritz C (1996) The uses of molecular phylogenies for conservation. In: Harvey PH, Leigh Brown AJ, Maynard Smith J, Nee S (eds) New uses for new phylogenies. Oxford University Press, Oxford, pp 203–214Google Scholar
  23. 23.
    Palumbi SR, Cipriano F (1998) Species identification using genetic tools: the value of nuclear and mitochondrial gene sequences in whale conservation. J Hered 89:459–464CrossRefPubMedGoogle Scholar
  24. 24.
    Palumbi SR, Cipriano F, Hare MP (2001) Predicting nuclear gene coalescence from mitochondrial data: the three-times rule. Evolution 55:859–868PubMedCrossRefGoogle Scholar
  25. 25.
    Povolny D (2002) Chrysomya albiceps (Wiedemann, 1918): the first forensic case in central Europe involving this blowfly (Diptera: Calliphoridae). Acta Univ Agric Silvic Mendel Brun 50:105–112Google Scholar
  26. 26.
    Ratcliffe ST, Webb DW, Weinzievr RA, Robertson HM (2003) PCR-RFLP identification of Diptera (Calliphoridae, Muscidae and Sarcophagidae)—a generally applicable method. J Forensic Sci 48:783–785PubMedGoogle Scholar
  27. 27.
    Rognes K, Paterson HEH (2005) Chrysomya chloropyga (Wiedemann, 1818) and C. putoria (Wiedemann, 1830) (Diptera: Calliphoridae) are two different species. Afr Entomol 13:49–70Google Scholar
  28. 28.
    Smith KGV (1986) A manual of forensic entomology. Cornell University Press, IthacaGoogle Scholar
  29. 29.
    Sperling FAH, Anderson GS, Hickey DA (1994) A DNA-based approach to the identification of insect species used for postmortem interval estimation. J Forensic Sci 39:418–427 (published erratum appears in letter from Wells JD, Sperling FAH (2000) J Forensic Sci 45:1358–1359)PubMedGoogle Scholar
  30. 30.
    Stevens J, Wall R (2001) Genetic relationships between blowflies (Calliphoridae) of forensic importance. Forensic Sci Int 120:116–123CrossRefPubMedGoogle Scholar
  31. 31.
    Stevens JR, Wall R, Wells JD (2002) Paraphyly in Hawaiian hybrid blowfly populations and the evolutionary history of anthropophilic species. Insect Mol Biol 11:141–148CrossRefPubMedGoogle Scholar
  32. 32.
    Swofford DL (1998) PAUP, phylogenetic analysis using parsimony (and other methods), Version 4. Sinauer, SunderlandGoogle Scholar
  33. 33.
    Swofford DL, Olsen GJ, Waddell PJ, Hillis DM (1996) Phylogenetic inference. In: Hillis DM, Moritz C, Mable BK (eds) Molecular systematics. Sinauer, Sunderland, pp 407–514Google Scholar
  34. 34.
    Vincent S, Vian JM, Carlotti, MP (2000) Partial sequencing of the cytochrome oxydase b subunit gene I: a tool for the identification of European species of blow flies for postmortem interval estimation. J Forensic Sci 45:820–823 (published erratum appears in letter from Wells JD, Sperling FAH (2000) J Forensic Sci 45:1358–1359)PubMedGoogle Scholar
  35. 35.
    Wagner DH (2000) Investigation of molecular population genetic substructure in forensically important flies. MS thesis, University of Alabama, BirminghamGoogle Scholar
  36. 36.
    Wallman JF, Donnellan SC (2001) The utility of mitochondrial DNA sequences for the identification of forensically important blowflies (Diptera: Calliphoridae) in southeastern Australia. Forensic Sci Int 120:60–67CrossRefPubMedGoogle Scholar
  37. 37.
    Wallman JF, Leys R, Hogendoorn K (2005) Molecular systematics of Australian carrion-breeding blowflies based on mitochondrial DNA. Invertebr Syst 19:1–15CrossRefGoogle Scholar
  38. 38.
    Wells JD (2000) Introduced Chrysomya Robineau-Desvoidy (Diptera: Calliphoridae) flies in northcentral Alabama. J Entomol Sci 35:91–92Google Scholar
  39. 39.
    Wells JD, Sperling FAH (1999) Molecular phylogeny of Chrysomya albiceps and C. rufifacies (Diptera: Calliphoridae). J Med Entomol 36:222–226PubMedGoogle Scholar
  40. 40.
    Wells JD, Sperling FAH (2001) DNA-based identification of forensically important Chrysomyinae (Diptera: Calliphoridae). Forensic Sci Int 120:109–114CrossRefGoogle Scholar
  41. 41.
    Wells JD, Introna FG, DiVella G, Campobasso CP, Hayes J, Sperling FAH (2001) Human and insect mitochondrial DNA analysis from maggots. J Forensic Sci 46:685–687PubMedGoogle Scholar
  42. 42.
    Wells JD, Lunt N, Villet MH (2004) Recent African derivation of Chrysomya putoria from C. chloropyga and mitochondrial DNA paraphyly of cytochrome oxidase subunit one in blowflies of forensic importance. Med Vet Entomol 18:445–448CrossRefPubMedGoogle Scholar
  43. 43.
    Zehner R, Amendt J, Schutt S, Sauer J, Krettek R, Povolny D (2004) Genetic identification of forensically important flesh flies (Diptera: Sarcophagidae). Int J Legal Med 118:245–247CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  1. 1.Department of BiologyWest Virginia UniversityMorgantownUSA
  2. 2.US Army Investigation LaboratoryForest ParkUSA

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