Molecular differentiation of Central European blowfly species (Diptera, Calliphoridae) using mitochondrial and nuclear genetic markers

Gilarriortua, Maite; Bordas, Marta Inés Saloña; Köhnemann, Stephan; Pfeiffer, Heidi; Pancorbo, Marian M. de

doi:10.1016/j.forsciint.2014.07.018

Cited by 27 publications

(12 citation statements)

References 36 publications

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“…In either case the addition of the nuclear gene ITS2 resolved the monophyly of the four species that COI alone did not support, and added resolution for uncertain groups with mtDNA genetic distances lower than 2%. These findings agreed with previous studies where the analysis of ITS2 resolved complex species delimitation ( GilArriortua et al, 2014 ; Song, Wang & Liang, 2008 ), however, not always addition of more genes resolved the monophyly of the sister species like the case of L. illustris and L. caesar , where, after analysis including six genes, the monophyly remain unresolved ( Sonet et al, 2012 ).…”

Section: Discussionsupporting

confidence: 92%

“…Given the serious implications of misidentification of forensic insects, an improved protocol for accurate identification is necessary. We propose using the nuclear internal transcribed spacer ITS2 as a second barcoding locus for taxonomic species determinations in calliphorids as suggested by GilArriortua et al (2014) . Although evaluations of ITS2 as unique identification marker have limitations for some taxa ( Agnarsson, 2010 ), several studies have shown the potential application of ITS2 for blowfly species identification ( Jordaens et al, 2013a ; Nelson, Wallman & Dowton, 2007 ; Nelson, Wallman & Dowton, 2008 , Song, Wang & Liang, 2008 ; Yusseff-Vanegas & Agnarsson, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

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DNA-barcoding of forensically important blow flies (Diptera: Calliphoridae) in the Caribbean Region

Yusseff-Vanegas

Agnarsson

2017

PeerJ

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Correct identification of forensically important insects, such as flies in the family Calliphoridae, is a crucial step for them to be used as evidence in legal investigations. Traditional identification based on morphology has been effective, but has some limitations when it comes to identifying immature stages of certain species. DNA-barcoding, using COI, has demonstrated potential for rapid and accurate identification of Calliphoridae, however, this gene does not reliably distinguish among some recently diverged species, raising questions about its use for delimitation of species of forensic importance. To facilitate DNA based identification of Calliphoridae in the Caribbean we developed a vouchered reference collection from across the region, and a DNA sequence database, and further added the nuclear ITS2 as a second marker to increase accuracy of identification through barcoding. We morphologically identified freshly collected specimens, did phylogenetic analyses and employed several species delimitation methods for a total of 468 individuals representing 19 described species. Our results show that combination of COI + ITS2 genes yields more accurate identification and diagnoses, and better agreement with morphological data, than the mitochondrial barcodes alone. All of our results from independent and concatenated trees and most of the species delimitation methods yield considerably higher diversity estimates than the distance based approach and morphology. Molecular data support at least 24 distinct clades within Calliphoridae in this study, recovering substantial geographic variation for Lucilia eximia, Lucilia retroversa, Lucilia rica and Chloroprocta idioidea, probably indicating several cryptic species. In sum, our study demonstrates the importance of employing a second nuclear marker for barcoding analyses and species delimitation of calliphorids, and the power of molecular data in combination with a complete reference database to enable identification of taxonomically and geographically diverse insects of forensic importance.

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Section: Discussionsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

DNA-barcoding of forensically important blow flies (Diptera: Calliphoridae) in the Caribbean Region

Yusseff-Vanegas

Agnarsson

2017

PeerJ

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“…In either case the addition of the nuclear gene ITS2 resolved the monophyly of the four species that COI alone did not support, and added resolution for uncertain groups with mtDNA genetic distances lower than 2%. This findings agreed with previous studies where the analysis of ITS2 resolve complex species delimitation (GilArriortua et al, 2014;Song, Wang & Liang, 2008), however, not always addition of more genes resolve the monophyly of the sister species like is the case of L. illustris and L. caesar where after analysis including six genes the monophyly remain unresolved (Sonet et al, 2012).…”

Section: Discussionsupporting

confidence: 92%

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Supplemental Information 2: Phylogenetic Relationship Within Calliphoridae Based on a Bayesian Analysis of Nucleotide Data From

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6Correct identification of forensically important insects, such as flies in the family Calliphoridae, 7 is a crucial step for them to be used as evidence in legal investigations. Traditional identification 8 based on morphology has been effective, but has some limitations when it comes to identify 9 immature stages of certain species. DNA-barcoding, using COI, has demonstrated potential for 10 rapid and accurate identification of Calliphoridae, however, this gene does not reliably

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“…The bootstrap values confirmed that molecular identification of L. cuprina is not complex and can be easily assayed with COI (figure 2), but identification of L. cuprina with conventional taxonomy was also congruent with molecular taxonomy. No monophyletic pattern using COI has been reported for L. cuprina [19,25,28,41]. DeBry et al [30] found a monophyletic group for L. cuprina in a vouchered collection of Lucilia species identified morphologically.…”

Section: Phylogenetic Analyses With Coi and 28s Regionsmentioning

confidence: 99%

“…Accuracy is mandatory in the identification of forensically important Lucilia species, but can be difficult because of similarities in their morphology [9]; only experts such as taxonomists and trained technicians can identify taxa accurately through extensive experience [10]. As a DNA approach was proposed for the identification of insect species for forensic issues [11] and for species identification [12], cytochrome oxidase I (COI) has been used world-wide for molecular identification of forensically important species [9,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27]such as Calliphoridae and Sarcophagidae, provides evidence for estimation of postmortem interval (PMI, including blow-fly genus Lucilia [28,29,30,31]. Some Lucilia species are so cosmopolitan that forensic entomology is specific to a locality; molecular studies are generally conducted in particular regions.…”

Section: Introductionmentioning

confidence: 99%

Identificación molecular mediante código de barras de DNA de moscas Lucilia (Diptera: Calliphoridae) recolectadas en Costa Rica

Sandoval-Arias¹,

Morales-Montero²,

Araya-Valcerde³

et al. 2020

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La identificación del género Lucilia es importante cuando se recolectan adultos o larvas en casos forenses porque el intervalo post-mortem puede ser estimado. En este trabajo se analizó morfológica y molecularmente un panel de especímenes de Lucilia (n = 42) recolectados en Costa Rica, de los cuales el 21% de las muestras corresponden a moscas criadas a partir de larvas tomadas de cadáveres humanos. Morfológicamente, los ejemplares fueron identificados como Lucilia cuprina [1] (33,3% de los ejemplares), Lucilia eximia [1] (33,3%), Lucilia purpurascens [2] (21,4%) o miembros del complejo Lucilia (11,9%). La identificación molecular, con las regiones de la citocromo oxidasa I (COI) o de la subunidad ribosomal 28S, sólo fue posible para el 43% de las muestras. La mayoría de las secuencias obtenidas con COI fueron concordantes con los análisis morfológicos, pero la región 28S carecía de suficiente resolución para identificar muestras al nivel de especies, excepto para L. cuprina. Juntos, los datos genéticos y morfológicos indican que L. cuprina y L. eximia fueron las especies más comúnmente encontradas; las moscas criadas a partir de larvas tomadas de cadáveres humanos correspondieron sólo a estas especies. Hasta donde sabemos, este trabajo es el primero en Costa Rica y en la región centroamericana en describir la entomofauna cadavérica del género Lucilia, lo cual es valioso para el desarrollo de potenciales aplicaciones forenses.

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Molecular differentiation of Central European blowfly species (Diptera, Calliphoridae) using mitochondrial and nuclear genetic markers

Cited by 27 publications

References 36 publications

DNA-barcoding of forensically important blow flies (Diptera: Calliphoridae) in the Caribbean Region

DNA-barcoding of forensically important blow flies (Diptera: Calliphoridae) in the Caribbean Region

Untitled

Identificación molecular mediante código de barras de DNA de moscas Lucilia (Diptera: Calliphoridae) recolectadas en Costa Rica

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