DNA barcoding of Lucilia blow flies (Diptera: Calliphoridae) collected in Costa Rica
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Published:
Feb 27, 2020
Keywords:
DNA barcoding, COI, 28S, Lucilia, Costa Rica
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Abstract
The identification of the genus Lucilia is important when adults or larvae are collected in forensic cases because the post-mortem interval might be estimated. In this work, we analyzed morphologically and molecularly a panel of Lucilia specimens (n = 42) collected in Costa Rica of which 21 % of the samples correspond to flies reared from larvae taken from human corpses. Morphologically, specimens were identified as Lucilia cuprina [1] (33.3 % of specimens), Lucilia eximia [1] (33.3 %), Lucilia purpurascens [2] (21.4 %) or members of Lucilia complex (11.9 %). Molecular identification with cytochrome oxidase I (COI) or 28S ribosomal subunit regions was possible for only 43 % of the samples. Most sequences obtained with COI were according to a morphological analysis, but the 28S region lacked sufficient resolution to identify samples to the level of species except L. cuprina. Together, genetic and morphological data indicate that L. cuprina and L. eximia were the most commonly found species; flies reared from larvae taken from human cadavers correspond to only these species. To our knowledge, this work is the first in Costa Rica and the Centro-American region to describe cadaveric entomofauna of Lucilia genus, which is valuable for the development of potential forensic applications.
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DNA barcoding of Lucilia blow flies (Diptera: Calliphoridae) collected in Costa Rica. (2020). Tecnología En Marcha Journal, 33(1), Pág. 99–110. https://doi.org/10.18845/tm.v33i1.5025
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References
[1] C.R.W. Wiedemann, “Brasilianische Zweiflgler”, Zool. Mag. 1(3): 40–56, 1819.
[2] F. Walker, “Descriptions, & c. of the Diptera”. In: Curtis, J., Haliday, A.H. & Walker, F. (Eds.), Descriptions, & c. of the insects collected by Captain P.P. King, R.N., F.R.S., in the survey of the Straits of Magellan. Transactions of the Linnean Society of London, 17(3): 331–359, 1836.
[3] J. Amendt et al., “Forensic entomology”, Naturwissenschaften, vol. 91, pp. 51–65, 2004. https://doi.org/10.1007/s00114-003-0493-5
[4] M.A.T. Marinho et al., “Molecular phylogenetics of Oestroidea (Diptera: Calyptratae) with emphasis on Calliphoridae: Insights into the inter-familial relationships and additional evidence for paraphyly among blowflies”, Mol Phylogenet Evol, vol. 65, pp. 840–854, 2012. https://doi.org/10.1016/j.ympev.2012.08.007
[5] S. Y. Cho et al., “DNA barcode library and its efficacy for identifying food-associated insect pests in Korea”. Bull. Entomol. Res. vol. 43, pp. 253–261, 2013. https://doi.org/10.1111/1748-5967.12034
[6] T. Withworth, “A revision of the neotropical species of Lucilia Robineau-Desvoidy”, Zootaxa, vol. 3810, pp. 1–76, 2014. http://dx.doi.org/10.11646/zootaxa.3810.1.1
[7] S. N. Kutty et al., “The Muscoidea (Diptera: Calyptratae) are paraphyletic: Evidence from four mitochondrial and four nuclear genes”, Mol Phylogenet Evol, vol. 49, pp. 639–652, 2008. https://doi.org/10.1016/j.ympev.2008.08.012
[8] J. Vargas and D.M. Wood, “Calliphoridae (blow flies)”, in Manual of Centro-American Diptera, B. Brown et al., M. (eds), Ottawa: NRC Research Press, pp. 1297–1304, 2010.
[9] B. K. Zajac et al., “Molecular identification and phylogenetic analysis of the forensically important family Piophilidae (Diptera) from different European locations”, Forensic Sci Int, vol. 259, pp. 77–84, 2016. https://doi.org/10.1016/j.forsciint.2015.12.024
[10] U. Jinbo et al., “Current progress in DNA barcoding and future implications for entomology”, Entomol Sci, vol. 14, pp. 107–124, 2011. https://doi.org/10.1111/j.1479-8298.2011.00449.x
[11] F.A.H. Sperling et al., “A DNA-Based Approach to the Identification of Insect Species Used for Postmorten Interval Estimation”, J Forensic Sci, vol. 39, pp. 418–427, 1994. https://doi.org/10.1520/JFS13613J
[12] P. D. N. Hebert et al., “Biological identifications through DNA barcodes”, Proc R Soc Lond [Biol],vol. 270, pp. 313–21, 2003. https://doi.org/10.1098/rspb.2002.2218
[13] J. F. Wallman and S.C. Donnellan, “The utility of mitochondrial DNA sequences for the identification of forensically important blowflies (Diptera: Calliphoridae) in southeastern Australia”, Forensic Sci Int, vol. 120, pp. 60–67, 2001. https://doi.org/10.1016/S0379-0738(01)00426-1
[14] J. D. Wells and F. A. H. Sperling, “DNA-based identification of forensically important Chrysomyinae (Diptera: Calliphoridae)”, Forensic Sci Int, vol. 120, pp. 110–115, 2001. https://doi.org/10.1016/S0379-0738(01)00414-5
[15] R. Zehner B. K. Zajac et al., “Genetic identification of forensically important flesh flies (Diptera: Sarcophagidae)” Int J Legal Med., vol. 118, pp. 245–247, 2004. https://doi.org/10.1007/s00414-004-0445-4
[16] W. Y. Chen et al., “Molecular identification of forensically important blow fly species (Diptera: Calliphoridae) in Taiwan”, J Med Entomol, vol. 41, pp. 47–57, 2004. https://doi.org/10.1603/0022-2585-41.1.47
[17] L. Cainé et al., “Genetic identification of forensically important Calliphoridae in Portugal”, Int Congr Ser, vol. 1288, pp. 846–848, 2006. https://doi.org/10.1016/j.ics.2005.09.153
[18] J. D. Wells and D. W. Williams, “Validation of a DNA-based method for identifying Chrysomyinae (Diptera: Calliphoridae) used in a death investigation”, Int J Legal Med., vol. 121, pp. 1–8, 2007. https://doi.org/10.1007/s00414-005-0056-8
[19] M. L. Harvey et al., “A global study of forensically significant calliphorids: Implications for identification”, Forensic Sci Int, vol. 177, pp. 66–76, 2008. https://doi.org/10.1016/j.forsciint.2007.10.009
[20] S. Reibe et al., “Molecular identification of forensically important blowfly species (Diptera: Calliphoridae) from Germany”, Parasitol Res, vol. 106, pp. 257–261, 2009. https://doi.org/10.1007/s00436-009-1657-9
[21] P. Boehme et al., “Molecular identification of carrion-breeding scuttle flies (Diptera: Phoridae) using COI barcodes”, Int J Legal Med., vol. 124, pp. 577–581, 2010. https://doi.org/10.1007/s00414-010-0429-5
[22] P. Boehme et al., “The use of COI barcodes for molecular identification of forensically important fly species in Germany”, Parasitol Res, vol. 110, pp. 2325–2332, 2012. https://doi.org/10.1007/s00436-011-2767-8
[23] K.A. Meiklejohn et al., “DNA-based identification of forensically important Australian Sarcophagidae (Diptera)”, Int J Legal Med., vol. 125, pp. 27–32, 2011. https://doi.org/10.1007/s00414-009-0395-y
[24] K. Jordaens et al., “Identification of forensically important Sarcophaga species (Diptera: Sarcophagidae) using the mitochondrial COI gene”, Int J Legal Med., vol. 127, pp. 491–504, 2013. https://doi.org/10.1007/s00414-012-0767-6
[25] M. GilArriortua et al., “Molecular differentiation of Central European blowfly species (Diptera, Calliphoridae) using mitochondrial and nuclear genetic markers”, Forensic Sci Int, vol. 242, pp. 274–282, 2014. https://doi.org/10.1016/j.forsciint.2014.07.018
[26] Y. Guo et al., “Identification of forensically important sarcophagid flies (Diptera: Sarcophagidae) in China based on COI and period gene”, Int J Legal Med., vol. 128, pp. 221–228, 2014. https://doi.org/10.1007/s00414-013-0923-7
[27] B. K. Zajac et al., “Application of DNA barcoding for identifying forensically relevant Diptera from northern Thailand”, Parasitol Res, vol. 115, pp. 2307–2320, 2016. https://doi.org/10.1007/s00436-016-4977-6
[28] J. D. Wells et al., “Phylogenetic analysis of forensically important Lucilia flies based on cytochrome oxidase I sequence: A cautionary tale for forensic species determination” Int J Legal Med., vol. 121, pp. 229–233, 2007. https://doi.org/10.1007/s00414-006-0147-1
[29] G. Sonet et al., “Why is the molecular identification of the forensically important blowfly species Lucilia caesar and L. illustris (family Calliphoridae) so problematic?”, Forensic Sci Int, vol. 223, pp. 153–159, 2012. https://doi.org/10.1016/j.forsciint.2012.08.020
[30] R. W. Debry et al., “DNA-Based Identification of Forensically Important Lucilia (Diptera: Calliphoridae) in the Continental United States”, J Forensic Sci, vol. 58, pp. 73–78, 2013. https://doi.org/10.1111/j.1556-4029.2012.02176.x
[31] M. GilArriortua et al., “Technical Note: “Mitochondrial and nuclear DNA approaches for reliable identification of Lucilia (Diptera, Calliphoridae) species of forensic interest from Southern Europe”, Forensic Sci Int, vol. 257, pp. 393–397, 2015. https://doi.org/10.1016/j.forsciint.2015.10.010
[32] J. Vargas “Distribución y morfología de adultos e inmaduros de moscas califóridas (Diptera: Calliphoridae) de importancia forense en Costa Rica”, Lic. Universidad de Costa Rica, 1999.
[33] R. Morales, Elaboración de una clave taxonómica para adultos y larvas de las especies de los géneros Lucilia y Hemilucilia (Diptera: Calliphoridae) de Costa Rica. Lic. Universidad de Costa Rica, 2014. http://hdl.handle.net/10669/29686
[34] T. Hall, “BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT”, Nucleic Acids Symp Ser, vol. 41, pp. 95–98, 1999.
[35] K. Tamura et al., “MEGA6: Molecular evolutionary genetics analysis version 6.0”, Mol Biol Evol, vol. 30, pp. 2725–2729, 2013. https://doi.org/10.1093/molbev/mst197
[36] R. C. Edgar “MUSCLE: Multiple sequence alignment with high accuracy and high throughput”, Nucleic Acids Res, vol. 32, pp. 1792–1797, 2004.
[37] J. Amendt et al., “Best practice in forensic entomology - Standards and guidelines”, Int J Legal Med., vol. 121, pp. 90–104, 2007. https://doi.org/10.1007/s00414-006-0086-x
[38] J. Macquart, “New or little known exotic diptera”, Volume two, 3rd part. Roret, Paris, 304 pp, 1844.
[39] J.W. Meigen, “Systematic description of the known European two-winged insects”, Fifth part. Schulz, Hamm, 412 pp, 1826.
[40] R.C. Shannon, “Synopsis of the American Calliphoridae (Diptera)”. Proceedings of the Entomological Society of Washington, 28, 115–139, 1926.
[41] R. W. Debry et al., “MtDNA-based identification of Lucilia cuprina (Wiedemann) and Lucilia sericata (Meigen) (Diptera: Calliphoridae) in the continental United States”, Forensic Sci Int, vol. 202, pp. 102–109, 2010. https://doi.org/10.1016/j.forsciint.2010.04.038
[42] M. GilArriortua et al., “Cytochrome b as a useful tool for the identification of blowflies of forensic interest (Diptera, Calliphoridae)”, Forensic Sci Int, vol. 228, pp. 132–136, 2013. https://doi.org/10.1016/j.forsciint.2013.02.037
[43] J. F. Wallman and S.C. Donnellan, “The utility of mitochondrial DNA sequences for the identification of forensically important blowflies (Diptera: Calliphoridae) in southeastern Australia”, Forensic Sci Int, vol. 120, pp. 60–67, 2001. https://doi.org/10.1016/S0379-0738(01)00426-1
[2] F. Walker, “Descriptions, & c. of the Diptera”. In: Curtis, J., Haliday, A.H. & Walker, F. (Eds.), Descriptions, & c. of the insects collected by Captain P.P. King, R.N., F.R.S., in the survey of the Straits of Magellan. Transactions of the Linnean Society of London, 17(3): 331–359, 1836.
[3] J. Amendt et al., “Forensic entomology”, Naturwissenschaften, vol. 91, pp. 51–65, 2004. https://doi.org/10.1007/s00114-003-0493-5
[4] M.A.T. Marinho et al., “Molecular phylogenetics of Oestroidea (Diptera: Calyptratae) with emphasis on Calliphoridae: Insights into the inter-familial relationships and additional evidence for paraphyly among blowflies”, Mol Phylogenet Evol, vol. 65, pp. 840–854, 2012. https://doi.org/10.1016/j.ympev.2012.08.007
[5] S. Y. Cho et al., “DNA barcode library and its efficacy for identifying food-associated insect pests in Korea”. Bull. Entomol. Res. vol. 43, pp. 253–261, 2013. https://doi.org/10.1111/1748-5967.12034
[6] T. Withworth, “A revision of the neotropical species of Lucilia Robineau-Desvoidy”, Zootaxa, vol. 3810, pp. 1–76, 2014. http://dx.doi.org/10.11646/zootaxa.3810.1.1
[7] S. N. Kutty et al., “The Muscoidea (Diptera: Calyptratae) are paraphyletic: Evidence from four mitochondrial and four nuclear genes”, Mol Phylogenet Evol, vol. 49, pp. 639–652, 2008. https://doi.org/10.1016/j.ympev.2008.08.012
[8] J. Vargas and D.M. Wood, “Calliphoridae (blow flies)”, in Manual of Centro-American Diptera, B. Brown et al., M. (eds), Ottawa: NRC Research Press, pp. 1297–1304, 2010.
[9] B. K. Zajac et al., “Molecular identification and phylogenetic analysis of the forensically important family Piophilidae (Diptera) from different European locations”, Forensic Sci Int, vol. 259, pp. 77–84, 2016. https://doi.org/10.1016/j.forsciint.2015.12.024
[10] U. Jinbo et al., “Current progress in DNA barcoding and future implications for entomology”, Entomol Sci, vol. 14, pp. 107–124, 2011. https://doi.org/10.1111/j.1479-8298.2011.00449.x
[11] F.A.H. Sperling et al., “A DNA-Based Approach to the Identification of Insect Species Used for Postmorten Interval Estimation”, J Forensic Sci, vol. 39, pp. 418–427, 1994. https://doi.org/10.1520/JFS13613J
[12] P. D. N. Hebert et al., “Biological identifications through DNA barcodes”, Proc R Soc Lond [Biol],vol. 270, pp. 313–21, 2003. https://doi.org/10.1098/rspb.2002.2218
[13] J. F. Wallman and S.C. Donnellan, “The utility of mitochondrial DNA sequences for the identification of forensically important blowflies (Diptera: Calliphoridae) in southeastern Australia”, Forensic Sci Int, vol. 120, pp. 60–67, 2001. https://doi.org/10.1016/S0379-0738(01)00426-1
[14] J. D. Wells and F. A. H. Sperling, “DNA-based identification of forensically important Chrysomyinae (Diptera: Calliphoridae)”, Forensic Sci Int, vol. 120, pp. 110–115, 2001. https://doi.org/10.1016/S0379-0738(01)00414-5
[15] R. Zehner B. K. Zajac et al., “Genetic identification of forensically important flesh flies (Diptera: Sarcophagidae)” Int J Legal Med., vol. 118, pp. 245–247, 2004. https://doi.org/10.1007/s00414-004-0445-4
[16] W. Y. Chen et al., “Molecular identification of forensically important blow fly species (Diptera: Calliphoridae) in Taiwan”, J Med Entomol, vol. 41, pp. 47–57, 2004. https://doi.org/10.1603/0022-2585-41.1.47
[17] L. Cainé et al., “Genetic identification of forensically important Calliphoridae in Portugal”, Int Congr Ser, vol. 1288, pp. 846–848, 2006. https://doi.org/10.1016/j.ics.2005.09.153
[18] J. D. Wells and D. W. Williams, “Validation of a DNA-based method for identifying Chrysomyinae (Diptera: Calliphoridae) used in a death investigation”, Int J Legal Med., vol. 121, pp. 1–8, 2007. https://doi.org/10.1007/s00414-005-0056-8
[19] M. L. Harvey et al., “A global study of forensically significant calliphorids: Implications for identification”, Forensic Sci Int, vol. 177, pp. 66–76, 2008. https://doi.org/10.1016/j.forsciint.2007.10.009
[20] S. Reibe et al., “Molecular identification of forensically important blowfly species (Diptera: Calliphoridae) from Germany”, Parasitol Res, vol. 106, pp. 257–261, 2009. https://doi.org/10.1007/s00436-009-1657-9
[21] P. Boehme et al., “Molecular identification of carrion-breeding scuttle flies (Diptera: Phoridae) using COI barcodes”, Int J Legal Med., vol. 124, pp. 577–581, 2010. https://doi.org/10.1007/s00414-010-0429-5
[22] P. Boehme et al., “The use of COI barcodes for molecular identification of forensically important fly species in Germany”, Parasitol Res, vol. 110, pp. 2325–2332, 2012. https://doi.org/10.1007/s00436-011-2767-8
[23] K.A. Meiklejohn et al., “DNA-based identification of forensically important Australian Sarcophagidae (Diptera)”, Int J Legal Med., vol. 125, pp. 27–32, 2011. https://doi.org/10.1007/s00414-009-0395-y
[24] K. Jordaens et al., “Identification of forensically important Sarcophaga species (Diptera: Sarcophagidae) using the mitochondrial COI gene”, Int J Legal Med., vol. 127, pp. 491–504, 2013. https://doi.org/10.1007/s00414-012-0767-6
[25] M. GilArriortua et al., “Molecular differentiation of Central European blowfly species (Diptera, Calliphoridae) using mitochondrial and nuclear genetic markers”, Forensic Sci Int, vol. 242, pp. 274–282, 2014. https://doi.org/10.1016/j.forsciint.2014.07.018
[26] Y. Guo et al., “Identification of forensically important sarcophagid flies (Diptera: Sarcophagidae) in China based on COI and period gene”, Int J Legal Med., vol. 128, pp. 221–228, 2014. https://doi.org/10.1007/s00414-013-0923-7
[27] B. K. Zajac et al., “Application of DNA barcoding for identifying forensically relevant Diptera from northern Thailand”, Parasitol Res, vol. 115, pp. 2307–2320, 2016. https://doi.org/10.1007/s00436-016-4977-6
[28] J. D. Wells et al., “Phylogenetic analysis of forensically important Lucilia flies based on cytochrome oxidase I sequence: A cautionary tale for forensic species determination” Int J Legal Med., vol. 121, pp. 229–233, 2007. https://doi.org/10.1007/s00414-006-0147-1
[29] G. Sonet et al., “Why is the molecular identification of the forensically important blowfly species Lucilia caesar and L. illustris (family Calliphoridae) so problematic?”, Forensic Sci Int, vol. 223, pp. 153–159, 2012. https://doi.org/10.1016/j.forsciint.2012.08.020
[30] R. W. Debry et al., “DNA-Based Identification of Forensically Important Lucilia (Diptera: Calliphoridae) in the Continental United States”, J Forensic Sci, vol. 58, pp. 73–78, 2013. https://doi.org/10.1111/j.1556-4029.2012.02176.x
[31] M. GilArriortua et al., “Technical Note: “Mitochondrial and nuclear DNA approaches for reliable identification of Lucilia (Diptera, Calliphoridae) species of forensic interest from Southern Europe”, Forensic Sci Int, vol. 257, pp. 393–397, 2015. https://doi.org/10.1016/j.forsciint.2015.10.010
[32] J. Vargas “Distribución y morfología de adultos e inmaduros de moscas califóridas (Diptera: Calliphoridae) de importancia forense en Costa Rica”, Lic. Universidad de Costa Rica, 1999.
[33] R. Morales, Elaboración de una clave taxonómica para adultos y larvas de las especies de los géneros Lucilia y Hemilucilia (Diptera: Calliphoridae) de Costa Rica. Lic. Universidad de Costa Rica, 2014. http://hdl.handle.net/10669/29686
[34] T. Hall, “BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT”, Nucleic Acids Symp Ser, vol. 41, pp. 95–98, 1999.
[35] K. Tamura et al., “MEGA6: Molecular evolutionary genetics analysis version 6.0”, Mol Biol Evol, vol. 30, pp. 2725–2729, 2013. https://doi.org/10.1093/molbev/mst197
[36] R. C. Edgar “MUSCLE: Multiple sequence alignment with high accuracy and high throughput”, Nucleic Acids Res, vol. 32, pp. 1792–1797, 2004.
[37] J. Amendt et al., “Best practice in forensic entomology - Standards and guidelines”, Int J Legal Med., vol. 121, pp. 90–104, 2007. https://doi.org/10.1007/s00414-006-0086-x
[38] J. Macquart, “New or little known exotic diptera”, Volume two, 3rd part. Roret, Paris, 304 pp, 1844.
[39] J.W. Meigen, “Systematic description of the known European two-winged insects”, Fifth part. Schulz, Hamm, 412 pp, 1826.
[40] R.C. Shannon, “Synopsis of the American Calliphoridae (Diptera)”. Proceedings of the Entomological Society of Washington, 28, 115–139, 1926.
[41] R. W. Debry et al., “MtDNA-based identification of Lucilia cuprina (Wiedemann) and Lucilia sericata (Meigen) (Diptera: Calliphoridae) in the continental United States”, Forensic Sci Int, vol. 202, pp. 102–109, 2010. https://doi.org/10.1016/j.forsciint.2010.04.038
[42] M. GilArriortua et al., “Cytochrome b as a useful tool for the identification of blowflies of forensic interest (Diptera, Calliphoridae)”, Forensic Sci Int, vol. 228, pp. 132–136, 2013. https://doi.org/10.1016/j.forsciint.2013.02.037
[43] J. F. Wallman and S.C. Donnellan, “The utility of mitochondrial DNA sequences for the identification of forensically important blowflies (Diptera: Calliphoridae) in southeastern Australia”, Forensic Sci Int, vol. 120, pp. 60–67, 2001. https://doi.org/10.1016/S0379-0738(01)00426-1