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Morphology and identification of first instars of the European and Mediterranean blowflies of forensic importance. Part II. Luciliinae
Abstract
First instars of Lucilia ampullacea Villeneuve, Lucilia caesar Linnaeus, Lucilia cuprina Weidemann, Lucilia richardsi Collin, Lucilia sericata Meigen and Lucilia silvarum Meigen (Diptera: Calliphoridae) are thoroughly documented with scanning electron microscopy images, light microscopy photographs and line drawings. The following morphological structures are documented: pseudocephalon, antennal complex, maxillary palpus, facial mask, cephaloskeleton, thoracic and abdominal spinulation, spiracular field, and posterior spiracles. New diagnostic features of the cephaloskeleton are presented and the spinulation of the abdominal segments is described. Earlier descriptions are summarized and major discrepancies with the current study are discussed. The present results allow for the clarification, correction and, especially, complementing existing information provided by numerous authors. The first instar larva of L. richardsi is described for the first time and an identification key to the first instars of European species of Lucilia Robineau-Desvoidy of forensic importance is presented.
... Larval terminology mainly follows Courtney et al. (2000) with a few modifications proposed by Cogley (1999), Szpila and Pape (2005), Szpila (2010), Szpila et al. (2008Szpila et al. ( , 2013 and Colwell et al. (2007). 'Oral plate' is a new term first proposed in this study (see further below). ...
... 'Oral plate' is a new term first proposed in this study (see further below). For antennomaxillary sensilla, the terms and numbering follow those for necrophagous calyptrate species (Szpila and Pape, 2005;Szpila et al., 2008Szpila et al., , 2013Szpila, 2010;Grzywacz et al., 2017), although the homology needs to be further corroborated. As the anterior spiracles of G. haemorrhoidalis, G. inermis, G. intestinalis, G. nasalis and G. pecorum have been well illustrated by Principato and Tosti (1988) and Principato, (1989), only the anterior spiracle of G. nigricornis is described complementarily. ...
... This structure is easily obscured by the retraction of the pseudocephalon into the first thoracic segment. Interestingly, the mouthhooks of the larvae are decorated with diversified ornamentations (polygonal plates, shallow pits, tile-like adornment), which is distinctive among calyptrates, all of which have smooth mouthhooks (Teskey et al., 1981;Ferrar, 1987;Szpila and Pape, 2005;Szpila et al., 2008Szpila et al., , 2013Szpila, 2010;De Filippis and Leite, 1998;Grzywacz et al., 2014). A similar condition (with tile-like adornment) is only found in the putative sister taxon, the genus Gyrostigma Brauer (rhinoceros stomach bot fly) (Erzinçlioglu, 1990). ...
Species of Gasterophilus are obligate parasites of equids and may induce severe, even lethal myiasis. However, identification of the third instar Gasterophilus larva at the species level is still problematic predominantly due to a shortage of diagnostic morphological features and incomplete molecular libraries. Testing the suitability of three different molecular markers showed that the traditional 650 bp barcode region near the 5′ terminus of the cytochrome c oxidase subunit I (COI) served as a better tool for species-level identification than a 663 bp region near the 3′ terminus of COI and a 554 bp region near the 5′ terminus of the large subunit ribosomal RNA. We found that barcoding discriminates G. intestinalis, G. nasalis, G. nigricornis and G. pecorum but not G. haemorrhoidalis and G. inermis. A comparative morphological study using scanning electron microscopy was conducted to promote the identification of the third instar larvae. Photographs of fresh mature third instar larvae are provided for all species, and the remarkable green body colour of third instar G. nigricornis is fully documented for the first time. Two morphological keys are provided, one is suitable for quick identification, and the other based on ultrastructural details is provided for further comparative morphological investigation. A new term ‘oral plate’ instead of ‘mandible’ was proposed for a pair of sclerites of uncertain homology emerging from the secondary mouth opening. Our data shows that DNA barcodes cannot replace morphology for identification of third instars of Gasterophilus species, and a scaffold is provided for an integrated taxonomic reference system, which will contribute to monitoring gasterophilosis for equid welfare and protection, and also facilitate further studies in functional anatomy, phylogenetic analyses and host-parasite co-evolutionary investigations of Gasterophilus.
... The calliphorid larvae are very similar and some authors have described the first and third instar of L. cuprina using light microscopy focusing on the cephalopharyngeal skeleton, which is http://dx.doi.org/10.1016/j.actatropica.2014.04.007 0001-706X/© 2014 Elsevier B.V. All rights reserved. the main structure visible using this technique (Sukontason et al., 2010;Szpila et al., 2013). However, light microscopy does not provide details of other characteristics that could have a diagnostic value (Liu and Greenberg, 1989;Mendonç a et al., 2010). ...
... More recently, Szpila et al. (2013) analyzed six species belonging to the genus Lucilia in Europe, but they focused their work on the first instar larvae. The general morphology of the larvae is very similar to our observations. ...
... However, due to the absence of studies comparing the larval morphology of the three species of the genus in Brazil, the doubts concerning the spinulation remain. In the L. cuprina collected in Amapá State, in the north of Brazil, the spinulation of segment A7 is incomplete, as related by Szpila et al. (2013). Although, the possibility of other arrangements could exist, since Wallman et al. (2005a,b) related the probable existence of cryptic species in L. cuprina. ...
The blowfly Lucilia cuprina is distributed worldwide and is a mechanical vector of pathogens. It can cause myiasis in humans and is strongly related to forensic entomology, as it is frequently found on human and animal corpses. However, most of the L. cuprina found on corpses are the immature stages of this fly. Correct identification is very important for forensic entomology but at present only the identification keys of adult L. cuprina are available. Thus, the aim of this paper was to describe and analyze the morphological characteristics of all larval instars and the puparia of L. cuprina using scanning electron microscopy (SEM).
... prefers shaded forest locations (Szpila et al. 2013b) similar to our findings during spring. Gennard (2007) and Vanin et al. (2008) mentioned this species on human remains. ...
... It is for the first time recorded in Lebanon, possibly occurring in all countries of the Middle East (Akbarzadeh et al. 2015). It is a facultative parasite and a well-known carrion breeder in the Oriental, Afrotropical, American, and Australian regions (Szpila 2010, Szpila 2013b. It was collected from hot and dry conditions but not yet detected in forensic cases or from carrion experiment in Europe (Szpila et al. 2013b). ...
... It is a facultative parasite and a well-known carrion breeder in the Oriental, Afrotropical, American, and Australian regions (Szpila 2010, Szpila 2013b. It was collected from hot and dry conditions but not yet detected in forensic cases or from carrion experiment in Europe (Szpila et al. 2013b). ...
Calliphoridae are among the first insects associated to decomposing animal remains. We have collected 1,841 specimens of three calliphorid genera: Calliphora, Lucilia, and Chrysomya, from different Lebanese localities as a first step in implementing a database of insects of forensic relevance for the country. Blow-flies are crucial for the estimation of the postmortem interval. DNA-based identification is a rapid and accurate method, often used for morphologically similar species, especially for immatures or incomplete specimens. In this study, we test the suitability of three genetic markers to identify adults and immature stages of calliphorids, viz., mitochondrial cytochrome c oxidase subunit I (COI) barcode, a region including partial sequences of mitochondrial Cyt-b-tRNAser-ND1, and second internal transcribed spacer (ITS2) region of nuclear ribosomal DNA. Forty Lebanese specimens of various developmental stages (egg, larva, wandering third instar, pupa, newly emerged adult, and mature adult) were identified among the three calliphorid genera: Calliphora, Lucilia, and Chrysomya, and compared with published sequences to confirm their specific assignation. Phylogenetic analyses showed the robustness of ITS2 and COI to identify calliphorids at species level. Nevertheless, ITS2 failed to discriminate Lucilia caesar (Linnaeus) (Diptera, Calliphoridae) from Lucilia illustris (Meigen) (Diptera, Calliphoridae), and COI had a similar issue with Lucilia sericata (Meigen) (Diptera, Calliphoridae) and Lucilia cuprina (Wiedemann) (Diptera, Calliphoridae). Thus, these two markers are complementary. This work contributes new nucleotide sequences for Lebanon. It is a first step in implementing a molecular database of forensic relevant insects for the country.
... The larvae were then washed in 5% acetic acid solution and rinsed in distilled water. The cephaloskeleton was dissected and slide-mounted in Hoyer's medium for light microscopy (Szpila et al. 2013a). Samples analyzed by scanning electron microscopy (SEM) were fixed in 70% alcohol, dehydrated in a series of increasing concentration of ethanol and subjected to drying to critical point using super-dry carbon dioxide in a Balzer critical point drier. ...
... Comparing with other first instar necrophagous species, besides a complete anterior spinose band until A6, the anterior spinose band of the anal division of P. pseudolyrcea is narrowly interrupted. H. segmentaria (incomplete bands from A2 to A7), Lucilia sericata (Meigen, 1826) (incomplete bands from A4 to A6), Lucilia cuprina (Wiedemann, 1830) (incomplete bands from A4 to A6), S. chlorogaster (incomplete bands from A5 to A7), C. albiceps (incomplete bands from A3 to A7), C. putoria (incomplete bands from A5 to A7) all have spinose bands that do not match P. pseudolyrcea pattern (Bonatto and Carvalho 1996, Thyssen and Linhares 2007, Florez and Wolff 2009, Szpila and Villet 2011, Szpila et al. 2013a. Besides it, P. pseudolyrcea differs from C. hominivorax, which have the anterior spinose band on the anal division complete (Laake et al. 1936. ...
... The cephaloskeleton, as the pattern of spines, is described for most of the necrophagous species and provides an important taxonomic character (Szpila et al. 2013a(Szpila et al. , b, 2014. The cephaloskeleton of the first instar of P. pseudolyrcea follows the same pattern of Calliphoridae. ...
A fundamental assumption of forensic entomology for estimating the postmortem interval is that insect species are accurately identified, which depends on diagnostic morphological characters. Larvae of the blow fly Paralucilia pseudolyrcea (Mello, 1969) (Diptera: Calliphoridae) were sampled from four corpses in the state of Paraná, Brazil, but despite the forensic importance of this species, morphological data for the identification of its larval instars are lacking, limiting its usefulness in such cases. Thus, the main goal of this study was to describe the larval instars of P. pseudolyrcea. The material was obtained from a colony established by larvae collected from a corpse of a murder case. Overall, the distribution of spines is a key character for identifying this species in the first, second and third instars. Other characteristics, such as the presence of an accessory oral sclerite, the small cirri, the number of lobes of the anterior spiracle and the morphology of posterior spiracles, separates P. pseudolyrcea from other necrophagous blow flies. The detailed morphological description provided here facilitates the identification of larval instars of P. pseudolyrcea and their differentiation from those of other calliphorid species.
... Among them are also descriptions, revisions, and keys useful for species identification of preimaginal stages. Articles with attempts to providing keys for identification of larval instars of European species of Lucilia were published by Schumann (1954Schumann ( , 1971, Szpila (2010), Szpila et al. (2013), and Velasquez et al. (2010). European species of Lucilia were also included in keys dedicated to other zoogeographical zones (Ishijima 1967;Knipling 1939;Liu and Greenberg 1989). ...
... Characters used in traditional keys are details of the cephaloskeleton, the distribution of spines on particular segments of the larval body, the position of papillae on the anal division, and details of posterior spiracles. Recently, the taxonomic value of some of these characters was critically revised especially in the context of identifying sister species like L. caesar and L. illustris (Szpila et al. 2013). ...
... We concentrated on the most apparent differences, however, in order to provide an easy template for quick determinations of individual larvae. For L. illustris and L. caesar-where species determination of larvae was reported to be difficult (Smith 1986;Szpila 2010;Szpila et al. 2013) or impossible (Schumann 1971) even with genetic methods (Sonet et al. 2012)-we found distinctive features in their respective MAS patterns. Larvae originating from different habitats and different mothers displayed accordant MAS patterns. ...
Species identification is generally assessed to be more difficult in larval stages than in adult forms. Especially closely related species such as Lucilia caesar and Lucilia illustris are difficult to identify. The aim of this study was to simplify species determination in Lucilia larvae for entomological and forensic purposes. Muscle attachment site (MAS) patterns were previously found to be a good tool for species determination in blowfly larvae. Here, distinctive MAS patterns are presented for European Lucilia ampullacea, L. caesar, L. illustris, L. richardsi, L. sericata, and L. silvarum. A joint pattern for the genus Lucilia is provided for a quick classification of a larva to the genus.
... Suitable agents for bleaching are, for example, methyl salicylate, KOH and Hoyer's medium (e.g. Semelbauer and Kozánek 2012; Velásquez et al. 2013; Szpila et al. 2013, 2014). The aim of our study is to demonstrate the utility of confocal laser scanning microscopy for studying the morphological characters of cyclorrhaphan Diptera larvae by taking advantage of the autofluorescent properties of the cephaloskeleton. ...
... The epistomal sclerite is equipped with four circular openings and fused anteriorly with the basal part of the labrum. The latter feature state was observed by Szpila et al. (2013); however, until recently, it has not been visualized exactly and Szpila et al. (2013) termed it the posterior expansion of the labrum, which is hump-shaped in Luciliinae. Grzywacz and Pape (2014) revealed that the presence of the epistomal sclerite in the first instar of Cyclorrhapha has been overlooked or confused by previous authors with only very few exceptions. ...
... The epistomal sclerite is equipped with four circular openings and fused anteriorly with the basal part of the labrum. The latter feature state was observed by Szpila et al. (2013); however, until recently, it has not been visualized exactly and Szpila et al. (2013) termed it the posterior expansion of the labrum, which is hump-shaped in Luciliinae. Grzywacz and Pape (2014) revealed that the presence of the epistomal sclerite in the first instar of Cyclorrhapha has been overlooked or confused by previous authors with only very few exceptions. ...
Larval morphology of flies is traditionally studied using light microscopy, yet in the case of fine structures compound light microscopy is limited due to problems of resolution, illumination and depth of field, not allowing for precise recognition of sclerites’ edges and interactions. Using larval instars of cyclorrhaphan Diptera, we show the usefulness of confocal laser scanning microscopy (CLSM) for studying the morphological characters of immature stages by taking advantage of the autofluorescent properties of cephaloskeleton structures. We compare data obtained from killed but unprepared larvae with those from larvae prepared by clearing according to two commonly used methods, either with potassium hydroxide or with Hoyer’s medium. We also evaluated the CLSM application for examining already slide-mounted larvae stored in museum collections and those freshly prepared. Our results indicate that CLSM and 3D reconstruction are excellent for visualizing small, compound structures of cylrorrhaphan larvae cephaloskeleton, if appropriate clearing techniques, i.e. the application of KOH, are used. Maximum intensity projection of confocal data sets obtained from material freshly prepared and that stored in museum collection does not differ. Because of this and the fact that KOH is commonly used as a clearing method to examine the cephaloskeleton of Diptera larvae, it is possible, and highly recommended, to use slides already prepared with this method for re-examination by CLSM. We conclude that CLSM application can be an invaluable source of data for studies of larval morphology of Cyclorrhapha by way of taxonomic diagnoses, character identification and improvement in characters homologization.
Electronic supplementary material
The online version of this article (doi:10.1007/s00436-014-4125-0) contains supplementary material, which is available to authorized users.
... Like that of the Luciliinae (Szpila et al., 2012a), knowledge of the morphology of first instars of European and Mediterranean Calliphorinae is unevenly distributed across the subfamily. Early larval instars of Ca. vicina have been described several times (Tao, 1927;Hall, 1948;Schumann, 1954;Zumpt, 1965;Erzinçlioglu, 1985;Liu & Greenberg, 1989;Szpila et al., 2008;Ubero-Pascal et al., 2012). ...
... Other steps in the preparation of material and documentation were identical to those described by Szpila et al. (2012aSzpila et al. ( , 2012b. Scanning electron microscopy (SEM) images of Ca. vomitoria were taken with a Hitachi S-4700 scanning electron microscope (Hitachi Ltd, Tokyo, Japan). ...
... 2(a) The posterior expansion of the labrum appears in the form of a hump-shaped protuberance and is readily visible in the lateral view (Fig. 3H). Mouth-hooks show only two or three dark sclerotized teeth (see Szpila et al., 2012a: fig. 9C, F, I, L, O, R) → (Luciliinae). ...
First instars of Calliphora vomitoria (Linnaeus) and Cynomya mortuorum (Linnaeus) (both: Diptera: Calliphoridae) are thoroughly documented with scanning electron microscopy images, light microscopy photographs and line drawings. Substantial new data are provided for the following morphological structures: pseudocephalon; antennal complex; maxillary palpus; facial mask; cephaloskeleton; thoracic and abdominal spinulation; spiracular field, and posterior spiracles. Earlier descriptions are summarized and major discrepancies with the current study are discussed. An identification key to first-instar larvae of the five European species of Calliphorinae of forensic importance is presented. However, it remains impossible to distinguish the first instars of Ca. vicina and Cy. mortuorum based on external morphology with the current level of knowledge.
... The immature forms collected from corpses are commonly preserved in ethanol for later identification. However, there is a need to rear these larvae until adults in order to identify and analyze all the immature stages The calliphorid larvae are very similar and some authors have described the first and third instar of L. cuprina using light microscopy focusing on the cephalopharyngeal skeleton, which is the main structure visible using this technique (Sukontason et al. 2010;Szpila et al. 2013). However, light microscopy does not provide details of other characteristics that could have a diagnostic value (Liu and Greenberg, 1989;Mendonça et al. 2010). ...
... More recently, Szpila et al. (2013) analyzed six species belonging to the genus Lucilia in Europe, but they focused their work on the first instar larvae. The general morphology of the larvae is very similar to our observations. ...
... However, due to the absence of studies comparing the larval morphology of the three species of the genus in Brazil, the doubts concerning the spinulation remain. In the L. cuprina collected in Amapá State, in the north of Brazil, the spinulation of segment A7 is incomplete, as related by Szpila et al. (2013). Although, the possibility of other arrangements could exist, since related the probable existence of cryptic species in L. cuprina. ...
The blowfly Lucilia cuprina is distributed worldwide and is a mechanical vector of pathogens. It can cause myiasis in humans and is strongly related to forensic entomology, as it is frequently found on human and animal corpses. However, most of the L. cuprina found on corpses are the immature stages of this fly. Correct identification is very important for forensic entomology but at present only the identification keys of adult L. cuprina are available. Thus, the aim of this paper was to describe and analyze the morphological characteristics of all larval instars and the puparia of L. cuprina using scanning electron microscopy (SEM).
... Despite the significant increase in morphological descriptions of immature stages of necrophagous species in recent years [9][10][11][12][13][14][15][16], well-documented and user-friendly identification keys are still missing. Consequently, obtaining adult specimens becomes essential either to confirm the initial identification of eggs or larvae, if established, or as the only means of identification. ...
... The literature search highlights a scarcity of identification keys for preimaginal stages of forensically important Diptera species [20][21][22][23]. Among the dipteran families, well-documented identification keys have been established for all instars of Calliphoridae [10,13,14,24,25], as well as for the third instar larvae of Piophilidae [26], Sarcophagidae [27] and Muscidae [28]. ...
There is a significant gap in the availability of comprehensive identification keys for the early larval stages of forensically important fly species. While well-documented identification keys exist for the third instar larvae, particularly for the Calliphoridae, Muscidae and Sarcophagidae families, there is a notable scarcity of keys for the first, except Calliphoridae, and the second instar larvae, with no such resources available for muscid species. The second instar larvae suffer the most from the lack of morphological descriptions and available identification keys. The Muscidae is one of the most frequently reported dipteran families of forensic importance colonising animal cadavers and human corpses. Nevertheless, descriptions of the morphology of their early instars remain scarce and limited to only a few species, thus their larval identification is challenging or impossible. Considering the
numerous challenges associated with studying small-sized entomological material, we tested whether it is feasible to identify muscid flies to the species or at least genus level based predominantly on the details of the cephaloskeleton. To overcome the obstacle of observing details of small sclerites, especially their shapes and interconnections, we effectively employed confocal laser scanning microscopy (CLSM) as a supplementary method for light microscopy (LM). This study provides an identification key for first and second instar larvae of forensically important muscid species from the western Palaearctic (Europe, North Africa, Middle East). The proposed key primarily utilises details of the cephaloskeleton with only addition of external morphology.
... total of 11 species of Calliphoridae [Bellardia vulgaris (Robineau-Desvoidy, 1830); Calliphora loewi Enderlein, 1903;Calliphora vicina (Robineau-Desvoidy, 1830); Calliphora vomitoria (Linnaeus, 1758); Chrysomya albiceps (Wiedemann, 1819); Lucilia ampullacea Villeneuve, 1922;Lucilia caesar (Linnaeus, 1758) (Rognes, 1991(Rognes, , 1997(Rognes, , 2002Verves, 2003;Civelek and Tezcan, 2005;Şabanoğlu and Sert, 2010;Çoban and Beyarslan, 2013;Szpila et al., 2013;Pekbey et al., 2016;Dawah et al., 2019). From these, the existence of L. coeruleiviridis in Turkey (Çoban and Beyarslan, 2013) is doubtful because of the Nearctic and Neotropical distribution of that species (Byrd and Castner, 2001;Whitworth, 2010). ...
... Until now, without any particular checklist, a total of 26 species (including L. coeruleiviridis and Rhiniidae) have been listed from Turkey (Rognes, 1991(Rognes, , 2002Verves, 2003;Civelek and Tezcan, 2005;Şabanoğlu and Sert, 2010;Çoban and Beyarslan, 2013;Szpila et al., 2013;Akbarzadeh et al., 2015). With our study's results, the number of species has risen to 31 in Turkey. ...
Despite the increasing importance of species richness of blowfly fauna and their environmental, medical, and agricultural importance in the world they are poorly studied in Turkey. This study was carried out in 2014 and 2015 to determine the distribution, abundance, and species richness of blowfly species in Isparta Province of Turkey. A total of 15 species (10 from Calliphoridae, 4 from Polleniidae, and 1 from Rhiniidae) were identified from 13 different localities. The adult specimens were obtained from areas of organic and decayed organic matters, waste and dumpsite, surrounding waste water deposits, and flowering plants. While all the species were new for Isparta, Bellardia tatrica (Enderlein, 1933), Calliphora subalpina (Ringdahl, 1931), Lucilia silvarium (Meigen, 1826), Melinda gentilis (Robineau-Desvoidy, 1830) and Pollenia griseotomentosa (Jacentkovsky, 1944) were determined as new records for the Turkish fauna. According to the study results, the highest levels of blowfly species richness will be mainly focused in the eastern part of Isparta Province. Chrysomya albiceps and Lucilia sericata were determined as the most common species in the study.
... In principal, it can be applied to 1st and 2nd instar larvae and still provide accurate results. The 1st instar larvae are particularly difficult to identify morphologically due to their size, and there are very few taxonomists able to identify to species level at this age [24,25]. ...
... sericata, C. vicina and C. vomitoria) in the 1st instar larval stage of the life cycle using cuticular hydrocarbon analysis. Although it is possible to identify species of Luciliinae [24], Chrysomyinae [25,28] and Calliphorinae [28] in the 1st instar larval stage, hydrocarbon analysis offers a complimentary technique which could provide further confirmation of identification. This study is a preliminary study to establish the proof of concept which when successful can then be applied to other more complicated species, such as Sarcophagidae and Muscidae. ...
... Aspoas, 1991;Awad et al., 2003;Boonchu et al., 2004;Klong-klaew et al., 2012;Liu and Greenberg, 1989;Mendoç a et al., 2008Mendoç a et al., , 2010Sanit et al., 2012Sanit et al., , 2013Singh et al., 2012;Sukontason K. et al., 2003Sukontason K. et al., , 2004aSukontason et al., 2001Sukontason et al., , 2002Sukontason et al., , 2003aSukontason et al., ,b, 2005Sukontason et al., , 2007Thyssen and Linhares, 2007), (2) works using ethanol as fixative and critical point drying (e.g. Alencar and Leite, 1992;Colwell et al., 2007;Draber-Mońko et al., 2009;Erzinç lioglu, 1989;Filippis and Leite, 1997;Grzywacz, 2013;Grzywacz and Pape, 2010;Grzywacz et al., 2011Grzywacz et al., , 2012Grzywacz et al., , 2013Kitching, 1976;Leite, 1988;Leite andLopes, 1987, 1989;Leite and Scott, 1999;Leite and Williams, 1989;Lopes and Leite, 1989;Szpila, 2003Szpila, , 2010Szpila and Pape, 2005a;Szpila and Villet, 2011;Szpila et al., 2012;Velásquez et al., 2013). Notwithstanding, protocol adaptations are usually frequent concerning either the type of the fixation chemical reagents -osmium tetroxide alone (Wells et al., 1999), formalin (Colwell and O'Connor, 2000), formalin-aceto-alcohol (Colwell et al., 1998;Grodowitz et al., 1982;Otranto et al., 2004), glutaraldehyde-formaldehyde (Paños et al., 2013;Ubero-Pascal et al., 2012), or the desiccation techniqueair drying after hexamethyldisilizane (HMDS) treatment (Paños et al., 2013;Semelbauer and Kozánek, 2012;Szpila, 2004;Szpila and Pape, 2005b, 2007, air-drying after butanal treatment (Feng and Liu, 2012). ...
... Awad et al., 2003;Boonchu et al., 2004;Singh et al., 2012;Sukontason K. et al., 2003;Sukontason et al., 2003aSukontason et al., ,b, 2005Sukontason et al., , 2008, than those fixed in ethanol and desiccated by critical point (e.g. Draber-Mońko et al., 2009;Filippis and Leite, 1997;Grzywacz et al., 2011;Leite, 1988;Leite andLopes, 1987, 1989;Szpila, 2003Szpila, , 2010Szpila and Pape, 2005a;Szpila and Villet, 2011;Szpila et al., 2012;Velásquez et al., 2013) or air-drying following HMDS treatment (e.g. Semelbauer and Kozánek, 2012;Szpila, 2004Szpila, , 2010Szpila and Pape, 2005b, 2007. ...
In forensic practice, the use of arthropod evidences to estimate the postmortem interval is a very good approach when the elapsed time from death is long, but it requires the correct identification of the specimens. This is a crucial step, not always easy to achieve, in particular when dealing with immature specimens. In this case, scanning electronic microscopy (SEM) can be useful, but the techniques used to preserve specimens in forensic practice are usually different from those used to prepare specimens for SEM studies. To determine whether forensic evidences preserving techniques are also compatible with SEM analysis, we have compared specimens of all the immature stages of Calliphora vicina Robineau-Desvoidy, 1830 (Diptera, Calliphoridae) preserved in 70% ethanol, with others prepared with aldehydic fixative techniques that are more appropriate for SEM studies. At the same time, two drying techniques have also been compared with both fixative techniques, the critical point drying and air-drying following with hexamethyldisilazane treatment (HMDS). Our results indicate that there are not basis against recommending the use of ethanol to preserve forensic entomological evidences and that both drying methods appear to offer good results for second and third instar larvae, although HMDS behaves better with eggs and pupae.
... In the larvae of the species that cause myiasis, those specialisations may reflect adaptations to parasitism (Jánošková et al., 2010;Szpila et al., 2014;Li et al., 2021); hence, larval morphological studies can provide novel and relevant information on the biology of those species, as well as on the pathogenesis and evolution of the different types of myiasis. Structures like the cuticular spines, the posterior spiracles and the cephalopharyngeal skeleton have classically been the focus of larval morphological studies of both obligatory and facultative myiasis agents, mostly due to their usefulness for species identification (e.g., Gómez-Fernández, 1958;Zumpt, 1965;Jánošková et al., 2010;Szpila et al., 2013Szpila et al., , 2014Szpila et al., , 2024. However, apart from the cephalopharyngeal skeleton, the anatomical study of internal organs and structures of myiasis-causing species has largely been overlooked, in spite of its potential for providing new insights into different biological and physiological aspects (Martín-Vega et al., 2021;Caleffe et al., 2024). ...
... The results presented for C. megacephala vindicate the concept. For Lucilia illustris and Lucilia Caesarwhere species determination of larvae was reported to be difficult (Smith, 1986;Szpila, 2010;Szpila et al., 2013) with genetic methods (Sonet et al., 2012), MAS pattern was found as a distinctive feature . These clusters of dots represent both genus and species-specific pattern in the larval stages, allowing for accurate species identification (Niederegger and Spieβ, 2012;Niederegger et al., 2013Niederegger et al., , 2015. ...
... The buccal hooks in the mouth parts of the first larval instar are extremely short while such structures are greatly sophisticated on both the second and third instar (Sandeman et al., 1987). The spines of the seventh abdominal segment were used to identify L. cuprina (Szpila et al. 2013). The ramifications number of spiracles is one of the greatest remarkable characteristics to differentiate between genera of Diptera and species (Costa et al. 2006). ...
... The buccal hooks in the mouth parts of the first larval instar are extremely short while such structures are greatly sophisticated on both the second and third instar (Sandeman et al., 1987). The spines of the seventh abdominal segment were used to identify L. cuprina (Szpila et al. 2013). The ramifications number of spiracles is one of the greatest remarkable characteristics to differentiate between genera of Diptera and species (Costa et al. 2006). ...
... All the examined calliphorid larvae were molecularly identified as L. caesar, a facultative myiasis-causing agent. This species is widely distributed throughout Europe in both natural and synanthropic ecosystems (Szpila et al., 2013); their larvae mainly feed on animal carcasses, but also on living tissues as occurred in this study. Myiasis by L. caesar are mostly reported in captive wildlife (Gao et al., 2021), and the first case in free ranging wildlife was reported in a wild boar from Italy in 2021 (Pezzi et al., 2021). ...
A total of 35 male roe deer (Capreolus capreolus) from Lugo province (north-western Spain) were examined for pedicle myiasis between 2020 and 2022. All these animals had died by different causes and were examined or preserved just after their death to avoid post-mortem infestations. After external and internal head inspection, five animals were diagnosed with severe myiasis at the basis of the antlers (14.29%; 95% confidence interval = 5.38-31.04). All the affected bucks presented a cutaneous wound (1.5-15 cm in diameter) around the pedicles with extensive tissue destruction, exposition of frontal and parietal bones, and massive infestation by dipteran larvae (2-12 mm long). Four of five roe deer showed whitish egg clusters adhered to the basis of the antlers. Moreover, in one animal, six larvae had penetrated the cranial cavity, and in two of them larvae were also found in the nasopharyngeal cavity. All the cases were recorded in summer (May-August). Morphological identification and subsequent molecular confirmation revealed that all animals were infested by different larval stages of Lucilia caesar (Diptera: Calliphoridae). This study represents the first report of pedicle myiasis in Spain. Since this disease is reported sporadically, the detection of five cases in a short period of time suggests an increase in the incidence of this myiasis.
... The interactive key was built using the software LUCID™ builder v. 3.6. The nomenclature used to describe the anatomical characters followed Erzinçlioğlu (1985), Shewell (1987), Courtney et al. (2000), and Szpila et al. (2013). Scale bars in millimeters were included in the images. ...
Calliphoridae (Insecta, Diptera), popularly known as blow flies or carrion flies, as most are necrophagous comprises one of the most relevant families of insects on forensics. Currently, more than 1500 blow fly species are known, the majority of which can be found in the Old World. In the Neotropics, it is estimated that there are approximately 20 species of forensic importance. In view of the taxonomic impediment associated with the identification of immatures, in this study, we present an interactive identification key for third instar larvae of necrophagous Calliphoridae species. This key includes 12 Neotropical blow fly species of forensic importance, more than 70 pictures, schemes, and definitions glossary and is free to access.
... This is the first case of myiasis caused by Lucilia caesar ever reported in a wild boar. This fly species, widely distributed in the Palearctic region [6,14] and common in Europe in synanthropic and natural ecosystems [15], is predominantly saprophagous [16][17][18]. Previous studies established that L. caesar was the dominant species among the necrophagous fauna in wooded areas of western Spain [19] and England [18,20], among necrophagous Diptera in rural grasslands of Southeast England [21] and in rural areas of Southern Italy [22]. ...
The first case of myiasis caused by Lucilia caesar (L.) (Diptera: Calliphoridae) in a wild boar, Sus scrofa L. (Artiodactyla: Suidae) is described. The myiasis occurred in October 2019 in Southern Italy and the identification of the agent was based on adult male morphology. The wild boar had a wound on its right side, near the neck, which was largely infested by larvae. The ecology, distribution and current literature status about cases of animal myiasis by this species is also included.
... It were taken from the various traps installed in the cage (barber and yellow traps) and also direct captures from the corpse. In the laboratory, insects were sorted and classifi ed using classifi cation keys [3,4], magnifying lenses and various tools used in the study of insects Table 1. Forensic medical entomology is concerned with studying insects related to a human body and one of its most important applications is to determine the time that has passed since death. ...
... However, through studies of the anatomy and physiology of the neurons present at the base of these sensilla, it was possible to determine that the Keilin's organs will give rise to the legs of adult insects (Lakes-Harlan et al. 1991). This structure is very similar between the species studied and when compared with the literature (Szpila et al. 2013(Szpila et al. , 2014. ...
Musca domestica (Linnaeus, 1758) is a muscoid species that is widespread throughout the world and acts as a mechanical vector of different enteropathogens primarily in underdeveloped countries. The adult and its immature forms are associated with decaying organic matter and can be seen visiting human corpses and animal carcass, and the larvae can also cause an infestation on human and animal wounds, feeding on the tissues. These characteristics make them have a forensic significance and, mainly, a great sanitary importance. This study aimed to analyze and describe morphological aspects of their immature stages of M. domestica, including the eggs, the first-, second-, and third-instar larvae, and the puparium using scanning electron microscopy (SEM). The eggs have standard format that all muscoid flies shared. The exochorion has some modified cells that were used for embryonic respiration. The first-instar larvae present two openings on the posterior spiracle as in the second-instar larvae, but with the last one, we can observe the anterior spiracle structure. The third-instar larvae, as in some other Muscidae species, have a posterior spiracular opening with a sinuous form that is located near the edge of the spiracular plate. The puparium morphology is equal to the third instar with a respiratory structure that helps the breathing during the metamorphosis process. This article supports the need for knowledge over the morphological characteristics of the immature forms of the muscoid Diptera (Linnaeus, 1758) at the same time helping with the correct identification of this insect phase.
... The environmental temperature was noted at each sampling using a Lascar EL-USB-2 data logger. Identification of necrophagous fly larvae was carried out in the laboratory using various identification keys (Wyss and Cherix, 2006;Szpila et al., 2008Szpila et al., , 2013. In order to study the life cycles of the flies, some of the collected larvae (LIII) were deposited in plastic boxes with a lid punctured with small holes to allow the air to enter. ...
The determination of benzodiazepines (carbamazepine and clobazam) in rabbit carcass tissues and larvae of three Calliphorid flies is described. After feeding the rabbits with lethal and toxic doses, samples of larvae and carcass tissues were studied. Residual drugs were determined using Ultra-high-performance liquid chromatography – quadrupole time-of-flight mass spectrometry (UHPLC / QTOF-MS). Benzodiazepines and its main active metabolites have been detected in the rabbit tissues at different retention times depending on the dosage used (lethal or toxic). A total of 1150 insects were collected and 800 larvae of the flies Chrysomya albiceps, Lucilia sericata and L. silvarum were used in the analysis. The presence of benzodiazepines in the rabbit tissues has been shown to typically affect the larval development cycle of the three necrophagous flies. Chrysomya albiceps larvae feed on drugs developed faster, while the development of L. sericata and L. silvarum larvae slowed. These results indicate that drugs have an impact on the life cycles of insects, which suggests that the presence of these substances is a factor that needs to be taken into account when estimating the post-mortem interval (PMI).
... e main characteristics used to classify the species of the larvae were its spiracles, peritrema, peritrema opening, tubers, mouth hooks, respiratory tracheas, cephalopharyngeal skeleton, and the number of segments (Figure 2). e morphological identification revealed that the larvae obtained from the patient belonged to the Calliphoridae family, and all were in larval stage II of their life cycle [9]. ...
Prevention of nosocomial myiasis, or hospital-acquired larvae infestation, should be an essential part of all hospital infection control programs. However, little is known about nosocomial myiasis, despite the extensive medical and psychological effects it has on patients and their families and the negative effects it has on hospitals’ reputation and finances. This report describes a case of nosocomial myiasis of a 13-year-old boy who was admitted to a pediatric intensive care unit for congestive heart failure, anemia, uremic encephalopathy, hypertension, and severe respiratory distress. Ten days after admission, the pediatrician and the nurse perceived an increase in the volume of the gingival mucosa of the upper buccal vestibule and the presence of fly larvae. The maggots were sent to the Instituto de Diagnostico y Referencia Epidemiologicos for identification and were found to be Lucilia sericata larvae. This report highlights the need to educate medical and paramedical personnel, as well as creation and implement protocols in hospitals to avoid nosocomial myiasis and improvement of general sanitation.
... Studied specimens originated from different localities and previous experiments, and were col- .The source of these arthropods were pigs, birds, and rats. Most of these specimens were imagos, which were previously identified morphologically through various authors and experts using standard keys and literature (3,(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44). The remaining specimens were identified to the order and/or family level only prior to Sanger Sequencing and in these cases, species identification was performed by reverse taxonomy using the SNSB, ZSM DNA barcode reference data on BOLD. ...
Throughout the years, DNA barcoding has gained in importance in forensic entomology as it leads to fast and reliable species determination. High‐quality results, however, can only be achieved with a comprehensive DNA barcode reference database at hand. In collaboration with the Bavarian State Criminal Police Office, we have initiated at the Bavarian State Collection of Zoology the establishment of a reference library containing arthropods of potential forensic relevance to be used for DNA barcoding applications. CO1‐5P’ DNA barcode sequences of hundreds of arthropods were obtained via DNA extraction, PCR and Sanger Sequencing, leading to the establishment of a database containing 502 high‐quality sequences which provide coverage for 88 arthropod species. Furthermore, we demonstrate an application example of this library using it as a backbone to a high throughput sequencing analysis of arthropod bulk samples collected from human corpses, which enabled the identification of 31 different arthropod Barcode Index Numbers.
... Outside of molecular techniques, morphological descriptions of the immature stages of blow flies are mandatory for species identification. As for blow flies in the genus Lucilia, although several SEM studies have been performed on immature stages; egg (Liu and Greenberg, 1989;Sukontason et al., 2007b;Sanit et al., 2013), larvae (Sandeman et al., 1987;Klong-klaew et al., 2012;Szpila et al., 2013a;Mendonça et al., 2014) and puparia , but no information pertaining to L. sinensis was found in the literature. We herein present the first description of all immature stages of L. sinensisegg, instars, and puparium. ...
Lucilia sinensis Aubertin (Diptera: Calliphoridae) is a blow fly species of potential forensic importance since adults are attracted to, and colonize, decomposing vertebrate remains. Blow fly larvae associated with human corpses can be useful evidence in forensic investigations; however, their use is dependent in most cases on proper species identification and availability of developmental data. For identification, morphological information on each life stage is traditionally used. We used scanning electron microscopy (SEM) to examine the ultrastructure of eggs, all instars, and puparia, of L. sinensis. The important characteristics used to differentiate L. sinensis from other species are provided. Distinctive features of the eggs are the slight widening median area extending almost the entire length. The last abdominal segment of the first instar bears elongated outer ventral tubercles along the rim of the last abdominal segment. These tubercles, as well as the well developed median and outer dorsal tubercles, are more prominent in the second and third instars. The surface integument of the tubercles is equipped with circular rows of microtrichia. Pairs of inner dorsal tubercle are absent. Each anterior spiracle is comprised of 9-12 papillae arrange in a single row in the second and third instars. As for the third instar, the dorsal spines between the first and second thoracic segments are delicate, narrow, small, and close together (as row or set). The peristigmatic tufts adjacent to the posterior spiracle of the third instar are moderately branches of short, fine hairs, but minute in puparia. In conclusion, the prominent outer ventral tubercle in all instars and puparia is a new diagnostic feature of L. sinensis and helpful in differentiating it from other Lucilia species that are forensically important. The description of immature L. sinensis in this study will be useful for forensic entomologists in countries where this species exists.
... This allowed for the composition of distinct species patterns for each of the six species. For L. illustris and L. caesar-where the species determination of larvae was reported to be difficult [38][39][40] or impossible [41], even with genetic methods [42]-MAS patterns were distinctive [37]. ...
Differences in growth rate of forensically important dipteran larvae make species determination an essential requisite for an accurate estimation of time since colonization of the body. Interspecific morphological similarities, however, complicate species determination. Muscle attachment site (MAS) patterns on the inside of the cuticula of fly larvae are species specific and grow proportionally with the animal. The patterns can therefore be used for species identification, as well as age estimation in forensically important dipteran larvae. Additionally, in species where determination has proven to be difficult—even when employing genetic methods—this easy and cheap method can be successfully applied. The method was validated for a number of Calliphoridae, as well as Sarcophagidae; for Piophilidae species, however, the method proved to be inapt. The aim of this article is to assess the utility of the MAS method for applications in forensic entomology. Furthermore, the authors are currently engineering automation for pattern acquisition in order to expand the scope of the method. Automation is also required for the fast and reasonable application of MAS for species determination. Using filters on digital microscope pictures and cross-correlating them within their frequency range allows for a calculation of the correlation coefficients. Such pattern recognition permits an automatic comparison of one larva with a database of MAS reference patterns in order to find the correct, or at least the most likely, species. This facilitates species determination in immature stages of forensically important flies and economizes time investment, as rearing to adult flies will no longer be required.
... Generally, the main obstacle in the use of morphological markers for identification of larvae of subgenus Liopygia is lack of solid comparative analysis. Studies of potentially useful characters, based on a long series of individuals and unified methodology, should solve this problematic situation as it was done for larval stages of some other difficult taxa (e.g., [24,25]). ...
In Mediterranean countries, Sarcophaga (Liopygia) crassipalpis , Sarcophaga (L.) argyrostoma, and Sarcophaga (L.) cultellata share the same ecological niche and can be responsible of myiasis. In this study, the main morphological characters of a larva found in a hospitalized woman were described and illustrated by light and SEM microscopy and the features discussed. Then, a fragment within the mitochondrial encoded cytochrome c oxidase subunit I (coxI) gene of ~735 bp was amplified and sequenced. The molecular investigation was necessary to confirm the species Sarcophaga (Liopygia) argyrostoma (99% of identity). Our findings showed that morphological descriptions of larvae of three Mediterranean species of Liopygia available in several papers might not be clear enough to allow for comparison and correct identification. Until results of reliable comparative studies of larvae of all three species will be available, the use of molecular tools is crucial, to avoid misleading or incomplete identification, and in particular when a myiasis becomes a legal issue.
... 28) possesses an elongated labrum (labelled as "median tooth"), without clear differentiation between an apical part and a basal part. Unfortunately, at this stage it is not possible to state that these differences between Madeira specimens and the original description result from interpopulation variation or inaccuracy of observation, as the larval material analyzed for the original description is unavailable (Szpila et al. 2013). Future studies on the variation of C. loewi larval morphology across the species distribution range will help to clarify this issue. ...
Knowledge on the taxonomic diversity and distribution of blowflies from the Madeira Archipelago is updated. New and interesting findings are reported for poorly studied islands and islets of this archipelago, together with a brief analysis of the diversity of Macaronesian Calliphoridae s. l. Seven blowfly species were collected during this study, including the first records of Calliphora vicina Robineau-Desvoidy, 1830, Chrysomya albiceps (Wiedemann, 1819), Lucilia sericata (Meigen, 1826), Pollenia rudis (Fabricius, 1794) and Stomorhina lunata (Fabricius, 1805) from Porto Santo, and of C. vicina, L. sericata and S. lunata from Desertas Islands. The presence of Calliphora loewi Enderlein, 1903 in Madeira Laurisilva forest is discussed and its first instar larva is redescribed, revealing important differences in relation to its original description. An identification key to the adult Madeiran blowflies is provided for the first time.
... The dorso-lateral edges of the functional mouth opening of all known first instar larvae of necrophagous blowflies are equipped with characteristic paired bunches of cirri/teeth (e.g. Sukontason et al., 2008;Ubero-Pascal et al., 2012;Szpila et al., 2013b), often called 'suprabuccal teeth' (e.g. Queiroz et al., 1997). ...
Light microscopy photographs, line illustrations and scanning electron microscopy micrographs are provided for first instar larvae of six Australian species of Chrysomya. All species have confirmed or potential in forensic investigations given their carrion-breeding habits. Morphology of the first instar larvae of Ch. nigripes, Ch. rufifacies, Ch. saffranea and Ch. varipes is revised, while larvae of Ch. incisularis and Ch. latifrons are described for the first time. The following morphological structures are documented: pseudocephalon, antennal complex, maxillary palpus, facial mask, thoracic and abdominal spinulation, spiracular field, posterior spiracles and cephaloskeleton. New diagnostic features of the cephaloskeleton and the spinulation of the abdominal segments are described. Verification of earlier descriptions revealed major discrepancies between published data, especially in the case of Ch. nigripes. The present results allow clarification, correction and, especially, complementation of the existing information provided by numerous authors. Finally, an identification key for first instar larvae of Australian necrophagous Chrysomya is presented.
... Only a few morphological keys exist, and their correct use requires experience and broad taxonomic knowledge (Kurahashi and Chaiwong 2013). Moreover, there are even fewer keys that allow classification at the larval stage (Szpila et al. 2013). The molecular identification of taxa is, hence, becoming more and more important in forensic entomology (Wells and Stevens 2008;Tarone et al. 2015). ...
In recent decades, forensic entomology has become a useful tool in criminal investigations all over the world. Species-specific identification of flies plays an important role in this field and is obligatory for accurate calculation of the post-mortem interval. However, not all important colonizers of a corpse can be identified by common morphological keys. Due to similar morphology and the lack of keys for some taxa, especially for immature stages, DNA barcoding has become more popular during the last recent years. This development is particularly important for countries like Thailand, in which forensic entomology is a newly developing research area and which faces several challenges such as a high biodiversity of fly species. The most commonly used barcoding region in forensic entomology, the mitochondrial cytochrome oxidase subunit 1 (coI) gene, as well as a 1000-bp-long region of the 28S nuclear rRNA gene, was used to analyze and establish the molecular barcodes of 13 different species of flies of forensic relevance in northern Thailand.
... While the situation is quite good for adult Diptera and Coleoptera, where useful keys for several sarcosaprophagous families can be applied [5][6][7], the situation is more complicated for the juvenile stages of these two very important orders. Here, only a few relevant keys exist [8,9], and even with such an appropriate key at hand, the correct species identification often requires special skills and expert knowledge of their morphology. ...
Species identification plays an important role in forensic entomology and is mandatory for an accurate calculation of the minimum post-mortem interval. Many important Diptera and Coleoptera taxa of the cadaver community can already be identified by common barcoding approaches, i.e., by sequencing a 658 bp region in the mitochondrial cytochrome c oxidase subunit I (coI) gene. Nevertheless, there is still a lack of reference barcodes for species, in particular, that can be found on cadavers at later decomposition stages. Flies of the family Piophilidae illustrate this gap of knowledge perfectly. Due to the fact that a reliable morphological identification key for the immature stages of this flies is still missing and the immature stages of many piophilids cannot be assigned to a certain species, there is need for additional tools to identify forensically relevant taxa. We collected adult piophilid specimens at 10 locations in five European countries: Spain (n = 3 locations), Germany (n = 3), Portugal (n = 2), Poland (n = 1) and Switzerland (n = 1). Apart from the coI barcoding region, we additionally analyzed a 398 bp long region of the nuclear elongation factor 1 alpha (ef1a) and subsequently established the molecular identifier for nine piophilid species. In addition, we present the molecular phylogeny of the examined taxa.
... The posterior spiracles are remarkable by showing perispiracular tufts with a slender, unbranched dorsal tuft, while the others are of a more typical palmate shape. This conÞguration is reminiscent of (and convergent to) that found in blow ßies of genus Lucilia Robineau-Desviody (Diptera: Calliphoridae) (Szpila et al. 2013), but the biological signiÞcance of this is unknown. ...
Flies in the family Sarcophagidae incubate their eggs and areknownto be ovoviviparous
(i.e., ovolarviparous), but a laboratory-maintained colony of Blaesoxipha plinthopyga (Wiedemann)
deposited clutches of viable eggs over 10 generations. A description of the egg and Þrst-instar larva
of this species is provided along with genetic data (genome size and cytochrome oxidase I sequences).
The egg is similar to previously described eggs of other Sarcophagidae but differs in the conÞguration
of the micropyle. In the Þrst-instar larva, the oral ridges are much more developed than has been
described for other species. B. plinthopyga has forensic importance, and the present descriptive
information is critical for proper case management.
... It is also worth mentioning that only few authors (e.g., Kozánek, 2011, 2012) have correctly identified the occurrence of an epistomal sclerite in the first instar larva of other Cyclorrhapha, and this structure has been variously described as the "base of labrum" (Szpila and Pape, 2005), "labrum supporting sclerite" (Ubero-Pascal et al., 2012), "process diverging from Abbreviations: an, antennal complex; as, anterior spiracle; ll, labial lobe; lo, labial organ; mp, maxillary palpus; ns1-2, first and second additional sensillum coeloconicum; or, oral ridges; sb1-3, sensillum basiconicum 1-3; sc1-3, sensillum coeloconicum 1-3; sub, suprabuccal teeth; vo, ventral organ. the parastomal bar" (Velasquez et al., 2013) or "posterior extension of the labrum" (Szpila et al., 2013(Szpila et al., , 2014. ...
Larval morphology is documented using both light and scanning electron microscopy for all three instars of the muscid fly Atherigona orientalis (Schiner), which is a species of known sanitary and forensic importance found in tropical and subtropical areas of all biogeographic regions. The unpaired sclerite in a form of a spicule is reported herein in the second and the third instar larvae. Occurrence of this sclerite was hitherto unknown in the second instar larvae of Muscidae and was only known from the third instar of several species, however not in a form of a spicule. Our study is the first report of the occurrence of the "sensory organ X" in all three larval instars of a species representing the family Muscidae. The bubble membrane, previously known only from third instar cyclorrhaphan larvae, is reported herein for the first time in the second instar. Characters allowing for discrimination of A. orientalis larvae from other forensically important Muscidae are summarized.
Myiasis is an infestation of the tissues of livinghumans and other vertebrates by the immature stages (larvae) of Diptera. According to the International Classification of Diseases, myiasis belongs to the morbidity classification ICD-10-CM, with the ICD code B87 (2025 edition). Myiasis is not mandatory to report, and through our experience, we aimed to raise awareness of this infestation among medical practitioners and the public. We discuss seven cases of myiasis diagnosed over five years (2019-2024) whose specimens (larvae) was submitted to the Institute for Microbiology and Parasitology, Medical Faculty - Ss. Cyril and Methodius University in Skopje, Republic of North Macedonia. The collected maggots were analyzed through macroscopic and microscopic examination. Morphological identification of the larvae was conducted using identification keys. Photographs were taken using an Olympus SZX9 and a Carl Zeiss Stemi 508 stereo microscope with an integrated high-resolution digital camera. Our findings confirmed an infestation by larvae from two autochthonous species, Sarcophaga argyrostoma and Lucilia sericata. We also identified an imported fly species, Cordylobia anthropophaga. The larvae can cause various symptoms depending on their relationship with the host and the anatomical location in the body. The outcome is related to comorbidities and the provision of appropriate, timely treatment. Medical practitioners should consider myiasis in their differential diagnosis. Nosocomial myiasis is a significant concern and requires special attention and preventive measures
Carrion-breeding flies are diverse with over 70 species in the Australasian/Oceanian region, predominantly from the families Calliphoridae and Sarcophagidae. These flies play crucial roles in ecosystems as primary decomposers, pollinators and food sources for a variety of predators and parasites. Given their unique ecologies and exceptional diversity, they have also proven to be particularly useful for human purposes in agriculture, in medicinal maggot therapy and in forensic entomology. Despite this, to date, there have been no comprehensive diagnostic tools developed for carrion-breeding flies in the Australasian region, which has hindered their use by non-experts in these applied fields. Here, we provide an
updated key for the identification of the adults of over 70 species of Australian and New Zealand flies known or suspected to breed in carrion. We also provide a review of the current state of knowledge regarding the biology and taxonomy of carrion-breeding flies in the Australasian region—summarising over a century of information regarding their distributions, available molecular data, biology, developmental data and the morphology of immature stages. Together, these resources will greatly improve the application of these species in forensics, agriculture, medicine and empirical research.
Microscopy is by far the most powerful tool in identification and study of the forensic flies. Our knowledge on fly morphology, reproductive parts, genitalia, male accessory glands, developmental stages of eggs (immature, pupa, larva), adult stages, salivary glands, antennal sensilla, and alimentary canals of flies are largely based on studies from light and scanning electron microscope that enables their identification, and some features like male genitalia find importance as identification keys in Calliphoridae. SEM studies have enabled identification of Chrysomya species from immature forms from corpses. In this chapter we highlight the advances of morphology-based studies in flies of forensic importance.
To date, there have been no studies on necrophagous fly populations on Hainan Island in China. Thus, we investigated the species composition of necrophagous flies as well as their geographic distribution on Hainan Island for the first time. Ten sites in different climatic regions across the island were sampled for 7 days per location from November 1, 2018 to February 28, 2019 and from July 1, 2019 to October 31, 2019. Bottle traps made of 1.5 L soft plastic bottles were used to trap necrophagous flies. The collected individuals were identified to species. The specimens represented 5 families and 28 species. Chrysomya megacephala (Fabricius, 1794) was the most dominant species, followed by Chrysomya rufifacies (Macquart, 1843), Hemipyrellia ligurriens (Wiedemann, 1830), Boettcherisca peregrine (Robineau-Desvoidy, 1830), Parasarcophaga dux (Thomson, 1868), Parasarcophaga misera (Walker, 1849), Synthesiomyia nudiseta (Wulp, 1883), and Ophyra chalcogaster (Wiedemann, 1924). The largest collection of flies was obtained in the semiarid region. Species richness was highest in the subhumid region and was higher in summer than in winter, but there were exceptions, such as L. hainanensis (Fan,1965), Boettcherisca formosensis (Lopes, 1961), and Muscina stabulans (Fallen, 1817). Fannia pusio (Wiedemann, 1830), and Boettcherisca formosensis (Lopes, 1961) were newly recorded species on Hainan Island. Of the necrophagous flies collected during the study, we propose several predominant species based on the criteria of distribution, occurrence frequency, and resource preference. Our results not only investigate necrophagous flies on Hainan Island but also accumulate data for criminal investigations in the region.
Geriatric Forensic Medicine and Pathology - edited by Kim A. Collins September 2020
L'entomologie médicolégale est la science qui applique les connaissances des insectes (et d'autres arthropodes) aux procédures civiles et pénales. C’est dans ce contexte que nous avons jugé intéressant d’apporter de nouvelles informations concernant la faune de Diptères liés à la décomposition cadavérique en Algérie, jusque là méconnue. Notre objectif est d'obtenir des données qui pourraient être utilisés pour les services concernés en l’occurrence la police scientifique.
Au cours de notre période d'étude nous avons mené une série d’expérimentations sur des
cadavres d’animaux (chiens et lapins), l'inventaire faunistique des espèces de Diptères
nécrophages récoltés aboutit au recensement de 18 espèces appartenant à 5 familles. La
famille des Calliphoridae est de loin la mieux représentée particulièrement avec les espèces Calliphora vicina, Lucilia sericata, Lucilia illustris et Calliphora vomitoria. Ces espèces offrent des outils qui contribuent à la définition du temps écoulé depuis la mort ou l’intervalle post mortem (IPM), qui est extrêmement précieux dans les enquêtes criminelles.
Afin d'examiner la relation qui existe entre la structure la faune des Diptères nécrophages et les différentes étapes de la décomposition cadavérique, des indices écologiques de structures et de compositions ainsi qu’une étude statistique composée d’une analyse de la variance et d’une analyse hiérarchique ont été utilisés.
OBJECTIVE: Aquatic forensic entomology is a branch of forensic science that applies the study of aquatic insects to criminal investigations, and is commonly used in death investigations
for estimating the post mortem submersion interval. However,
there is no study in the field of aquatic forensic entomology in
Turkey although there are deaths reported as found in water.
METHODS: In the province of Aksaray (city), two pigs’ heads
of Sus scrofa domestica (domestic pig) (Erxleben, 1777) species
in cages were placed into two different drainage channels for
36 days for the purpose of attracting and identifying forensically
significant aquatic insects. The insects on the carcasses were
collected to identify the species, and decay phases were report
RESULTS: The taxa that were collected from the first test
animal included 9 Diptera species from the families Calliphoridae, Muscidae, Fanniidae, Ephydridae and Chironomidae; 5 Coleoptera species and a subspecies from the families Histeridae,
Staphylinidae, Dytiscidae and Helophoridae; and one Gerridae
species from the family Hemiptera. The taxa identified in the
second test animal included 7 Diptera species from the families
Calliphoridae, Muscidae and Chironomidae; 4 Coleoptera species from the families Nitidulidae, Dytiscidae and Helophoridae;
one Hemiptera species from the family Corixidae; and one Odonata species from the family Libellulidae.
CONCLUSION: This is the first study carried out in Turkey
regarding the identification of forensically substantial aquatic
insects, and the detected taxa have been recorded for the first
time in terms of forensic aquatic entomology in our country
This represents one of several sections of "A Bibliography Related to Crime Scene Interpretation with Emphases in Geotaphonomic and Forensic Archaeological Field Techniques, Nineteenth Edition" (The complete bibliography is also included at ResearchGate.net.). This is the most recent edition of a bibliography containing resources for multiple areas of crime scene, and particularly outdoor crime scene, investigations. It replaces the prior edition and contains approximately 10,000 additional citations. As an ongoing project, additional references, as encountered, will be added to future editions.
The collection and analyses of insects, or invertebrates, from crime scenes is generally well known among homicide investigators and death scene investigators. It has been the compiler’s experience, however, that the actual practice of such collection, outside the presence of a forensic entomologist, is still overlooked or avoided. Often, an attitude prevails that that level of information is not necessary given the investigator’s knowledge of when an abduction took place, or a subject’s confession. In other situations, collections are not made simply because the investigators are not properly equipped with tools, chemicals, and packaging materials to collect and kill samples, or are not sure of what to do with, or how to store, live specimens. Entomological evidence is unique in that it is, in most criminal investigations, the only type of non-human evidence consisting of living, moving species. It is the hope of the compiler that this section will offer some answers toward appropriate collection procedures and equipment which are not expensive, do not involve a lot of time, or the need for additional manpower. The proper collection of entomological samples combined with accurate spatial, temporal, and environmental data, can yield valuable information toward determining postmortem intervals (Taphonomy - Decomposition and Time Since Death). Subject/Witness statements might be supported or disproved. Works such as Catts and Haskell (1990), and Lord and Burger (1983) have become standards in the field of forensic entomological procedures. In recent years compilations such as that by Byrd and Castner (2010) have demonstrated the increasing interest in forensic applications of a science which otherwise serves advancements in health and agriculture. Amendt, et al. (2007) offer "Standards and Guidelines" for this field of study. Entomology, as a means of determining post-mortem interval, continues to be scrutinized. This is not a bad thing. Any validation, clarification, or revocation of a forensic technique benefit crime scene interpretation.
Many of the works below include laboratory analyses and research. That research goes beyond addressing the timing of a death or deposition (post-mortem interval) to toxicological determinations, interpretations of death scene versus depositional environments, et cetera. Like virtually every category in this bibliography, the study of entomology is contingent upon so many factors at a scene that an understanding of other disciplines is a neccessity. Obviously, taphonomy and pathology are directly related to insect activity on discovered remains. Environmental characteristics such as soils and plants, as well as body position either by accident or intentional, could influence the impact of insects in peri- and post-mortem activity. Again, the reader is refered to other categories such as Taphonomy, Geoarchaeology and Soil Science, and Criminal and Cultural Behavior. (2552 citations)
Forensic entomology in Poland: 1) Turn of the Nineteenth and Twentieth Century, 2) Twentieth Century, 3) Twenty-First Century, 4) Training courses and conferences in forensic entomology, 5) Ongoing projects, 6) Use of forensic entomology in legal investigations
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.
Approved standards and practices are necessary for forensic entomology because its application must be reliable in court. Forensic entomology requires a template for collecting, analyzing, and reporting its evidence using common minimum standards and best practices that can be defended in court. This chapter provides a protocol modeled on medicocriminal cases that is easily modified for urban, stored-product, or environmental forensic cases. It is based on the forensic process, starting with collection of evidence at an investigation site, and culminating in the documentation of the forensic findings.
Calliphoridae are found in a variety of habitats, showing preference for human environments, a phenomenon known as synanthropy. The medical, ecological and sanitary importance of these flies resides in their feeding and development habits, which occur in excrement, garbage, decomposing organic matter and animal carcasses. In addition, they are mechanical vectors of pathogens and causal agents of myiasis. The objective of this study was to determine the synanthropic index (SI) of adult Calliphoridae in La Pintada-Antioquia, Colombia, through simultaneous monthly samplings in three environments (urban, rural, and forest) using van Someren Rydon traps baited with human excrement, decomposing chicken viscera, fish and onion. Four traps (one for each bait) were placed in each environment for 48 hours each month, making collections every 12 hours. A total of 2406 individuals was captured belonging to 10 species. Chysomya megacephala (SI = + 96.47) showed the strongest preference for dense human settlements. Cochliomyia macellaria (SI = + 78.1), Lucilia eximia (SI = + 74,94) and Chysomya albiceps (SI = + 59.12) had preferences for human settlements (eusynanthropic). Hemilucilia semidiaphana (SI = -50) showed a preference away from human settlements (asynanthropic). Cochliomyia hominivorax, Chloroprocta idioidea, Lucilia cuprina, Hemilucilia segmentaria and Paralucilia pseudolyrcea were collected in small numbers, and their SI could not be determined. The monthly abundance and the efficiency of the attractiveness to baits are discussed according to their medical and sanitary importance. Paralucilia pseudolyrcea is recorded for first time in Colombia. Synanthropic species could be involved in the transmission of pathogens in this locality.
The occurrence of blowflies with public health importance (four species of the genus Calliphora Robineau-Desvoidy, 1830, nine species of the genus Lucilia Robineau-Desvoidy, 1830 and one species of the genus Protophormia Townsend, 1908) was studied in the city of Brno and a village Ketkovice near Brno. A total of 2781 imagoes (1926 from Brno and 855 from Ketkovice) were obtained from both study areas by sweeping, trapping or rearing from eggs. Calliphora uralensis, C. vicina, C. vomitoria, Lucilia ampullacea, L. bufonivora, L. caesar, L. illustris, L. pilosiventris, L. regalis, L. sericata, L. silvarum and Protophormia terraenovae were found in both study areas. C. loewi and L. richardsi were caught only in Ketkovice and Brno, respectively. L. sericata was the dominant species in densely populated places in both areas. L. caesar was the dominant species in forests and one forest park in Brno and in the forests around Ketkovice. Although L. bufonivora is considered to be a rare species, it was not rare and it was the most numerous in forests and recreation areas near water reservoir and river banks.
The objective of this work was to describe the morphology and the developmental time of Chrysomya putoria larvae, a fly of public health and forensic importance. The samples were collected at 2 hr intervals, during the first 56 hr and at 12 hr intervals thereafter, until all mature larvae had abandoned the diet. The total developmental time for Chrysomya putoria larvae at 27 ºC was 92 hr, with a mean length of 13.42 mm. The larvae presented unique morphological characteristics which differentiated them from other exotic and autochthonous species found in Brazil. The cephalopharyngeal skeleton of the third instar larvae is robust and sclerotized; the mouth hook is completely pigmented and curved; the posterior end is wider than the anterior, being slightly pointed. The accessory oral sclerit is absent and the dorsal and ventral horns have areas with less pigmentation. It is possible to visualize an area with no pigmentation adjacent to the horns, outstanding in size near the ventral corn with a well delimited medial incision. The anterior spiracles presented 10 to 11 ramifications, open peritrema and the presence of a spiracular bud from the second instar on.
Morphology and duration of the post-embryonary development of Chrysomya megacephala (Diptera: Calliphoridae) in laboratory conditions. The morphology and developmental time of the post-embryonic stage of Chrysomya megacephala (Fabricius) reared in laboratory on a beef diet, is described and compared to those of other blow-flies species. The parameters analyzed were the duration of the larval stages and the morphology. Larvae were fixed and preserved at 2 hours intervals, from larval eclosion until the 50th hour of life, in order to determine the duration of each stage. The total period of larval development in C. megacephala was 98 hours and their mean length was 15.51 mm. This species presents a necrobiontophagous behavior and has sanitary and medico-legal importance, being used for estimating postmortem interval in criminal investigations.
Calliphoridae are found in a variety of habitats, showing preference for human environments, a phenomenon known as synanthropy. The medical, ecological and sanitary importance of these flies resides in their feeding and development habits, which occur in excrement, garbage, decomposing organic matter and animal carcasses. In addition, they are mechanical vectors of pathogens and causal agents of myiasis. The objective of this study was to determine the synanthropic index (SI) of adult Calliphoridae in La Pintada-Antioquia, Colombia, through simultaneous monthly samplings in three environments (urban, rural, and forest) using van Someren Rydon traps baited with human excrement, decomposing chicken viscera, fish and onion. Four traps (one for each bait) were placed in each environment for 48 hours each month, making collections every 12 hours. A total of 2406 individuals was captured belonging to 10 species. Chysomya megacephala (SI = + 96.47) showed the strongest preference for dense human settlements. Cochliomyia macellaria (SI = + 78.1), Lucilia eximia (SI = + 74,94) and Chysomya albiceps (SI = + 59.12) had preferences for human settlements (eusynanthropic). Hemilucilia semidiaphana (SI = -50) showed a preference away from human settlements (asynanthropic). Cochliomyia hominivorax, Chloroprocta idioidea, Lucilia cuprina, Hemilucilia segmentaria and Paralucilia pseudolyrcea were collected in small numbers, and their SI could not be determined. The monthly abundance and the efficiency of the attractiveness to baits are discussed according to their medical and sanitary importance. Paralucilia pseudolyrcea is recorded for first time in Colombia. Synanthropic species could be involved in the transmission of pathogens in this locality.
Carrion-breeding blowflies have substantial ecological and forensic importance. Because morphological recognition of their immatures is difficult, sequencing of the mtDNA of these flies may assist with their identification. Molecular phylogenetic analysis based on DNA sequences can also clarify evolutionary relationships. In this study, the mitochondrial genes CO1, CO2, ND4 and ND4L were sequenced for 34 species of blowflies, among which are almost all species known or suspected to breed in carrion in Australia. The resulting sequences were analysed using parsimony and maximum-likelihood Bayesian techniques. The results showed that the combination of these four genes should identify most species reliably, although some very closely related taxa could still be misdiagnosed. The data also helped clarify the life histories of Calliphora centralisMalloch, 1927, C. fuscofemorataMalloch, 1927 and C. gilesiNorris, 1994, which have hitherto only been suspected carrion breeders, and revealed that the current subgeneric assignment of taxa within Calliphora Robineau-Desvoidy, based on morphology, requires revision. Unexpectedly, both Chrysomya rufifacies (Macquart, 1843) and Lucilia cuprina (Wiedemann, 1830) were paraphyletic; each probably comprises two distinct species. The application of a molecular-clock approach to the study of the evolutionary divergence of the carrion-breeding blowflies suggests that the speciation of at least the endemic Australian taxa may have been the result of increasing aridification in Australia during the last five million years.
Due to the presence of an adult dead dark-winged fungus gnat (Bradysia sp., Sciaridae), a fly known to live on the borders of forests, on a corpse that was found on the edge of a forest in Western Germany, we were asked during the trial if we could define what the edge of a forest is. Using two piglets as models, we determined parameters to approach such a definition. The most obvious difference between the inside and the border of the forest was the outside temperature, and the inside temperature of the decomposing piglets, which was higher in the piglet at the border. Over all, 957 individual flies of 56 species of 12 families (Anthomyiidae, Calliphoridae, Drosophi- lidae, Dryomyzidae, Fanniidae, Heleomyzidae, Lauxaniidae, Muscidae, Piophilidae, Phoridae, Sep- sidae, Sphaeroceridae) were found. Specimens of the Drosophilidae and Sepsidae were found exclusively at the edge of the forest and those of the Lauxaniidae, Phoridae, and Sphaeroceridae inside the wood. 36% of the species were found exclusively on the corpse on the border of the forest whilst 10% of the species were exclusively found on the corpse inside of the forest. Several dipteran species showed a clear preference for the border of the forest (Calliphora vicina, Lucilia sericata, Fannia canicularis, Hydrotaea ignava, Liopiophila varipes, Parapiophila vulgaris, Stearibia nigriceps), or for the inside of the woods (Neuroctena analis, Fannia nigra). Succession waves were represented by proportion of insect groups, depending on the location of the piglet. Large numbers of heleo- myzids (especially Neoleria sp. 2) were found during the post-decay stage on both corpses. Lucilia ampullacea (Calliphoridae) arrived very early on the habitats, whilst Lucilia caesar arrived 1-2 days later. This is of relevance for post mortem interval analysis. Succession waves were also repre- sented by the relative numbers of adult individuals on the corpses. Here, the relative decrease of calliphorid individuals compared to the increase of heleomyzids, piophilids and dryomyzids was one of the most prominent markers. The decay stages showed a marked difference in the fauna between the two habitats. This was observed not only in the common but also the rare species (Rényis' diversity). The decomposing piglet on the border was inhabited by a significantly higher number of individuals (576 vs. 381) as well as species (50 vs. 36) of Diptera compared to the piglet inside of the forest. The overall differences in sexes were 23 %/301 & (Calliphoridae), 4 %/89 & (Muscidae), 1 %/87 & (Fanniidae), 116 %/41 & (Piophilidae), and 18 %/238 & (Heleomyzidae). *Minor parts of this article were presented as lectures at the meeting of the EAFE (European Association for Forensic Entomology), Lausanne, Switzerland, April 27-29, 2005, and at the 6 th
The dipteran clade Calyptratae is comprised of approximately 18 000 described species (12% of the known dipteran diversity) and includes well-known taxa such as houseflies, tsetse flies, blowflies and botflies, which have a close association with humans. However, the phylogenetic relationships within this insect radiation are very poorly understood and controversial. Here we propose a higher-level phylogenetic hypothesis for the Calyptratae based on an extensive DNA sequence dataset for 11 noncalyptrate outgroups and 247 calyptrate species representing all commonly accepted families in the Oestroidea and Hippoboscoidea, as well as those of the muscoid grade. DNA sequences for genes in the mitochondrial (12S, 16S, cytochrome c oxidase subunit I and cytochrome b) and nuclear genome [18S, 28S, the carbamoyl phosphate synthetase region of CAD (rudimentary), Elongation factor one alpha] were used to reconstruct the relationships. We discuss problems relating to the alignment and analysis of large datasets and emphasize the advantages of utilizing a guide tree-based approach for the alignment of the DNA sequences and using the leaf stability index to identify ‘wildcard’ taxa whose excessive instability obscures the phylogenetic signal. Our analyses support the monophyly of the Calyptratae and demonstrate that the superfamily Oestroidea is nested within the muscoid grade. We confirm that the monotypic family Mystacinobiidae is an oestroid and further revise the composition of the Oestroidea by demonstrating that the previously unplaced and still undescribed ‘McAlpine’s fly’ is nested within this superfamily as a probable sister group to Mystacinobiidae. Within the Oestroidea we confirm with molecular data that the Calliphoridae are a paraphyletic grade of lineages. The families Sarcophagidae and Rhiniidae are monophyletic, but support for the monophyly of Tachinidae and Rhinophoridae depends on analytical technique (e.g. parsimony or maximum likelihood). The superfamilies Hippoboscoidea and Oestroidea are consistently found to be monophyletic, and the paraphyly of the muscoid grade is confirmed. In the overall relationships for the calyptrates, the Hippoboscoidea are sister group to the remaining Calyptratae, and the Fanniidae are sister group to the nonhippoboscoid calyptrates, whose relationships can be summarized as (Muscidae (Oestroidea (Scathophagidae, Anthomyiidae))).
In Europe larvae of blowflies are the main group of insects responsible for decomposition of exposed vertebrate remains, including
the human body. This determines their high forensic importance and frequent application for estimation of PMI. The importance
of proper identification of insects collected in forensic cases and experiments to the species level is underlined by all
manuals of forensic entomology (e.g. Smith 1986; Byrd and Castner 2001; Greenberg and Kunich 2002). Especially difficult is
the identification of the larval stages, where breeding to the adult stage or DNA-based methods are recommended. Fortunately,
the available knowledge of the morphology of third instars of Calliphoridae is sufficiently good to allow the preparation
of a complete identification key for at least all European species of forensic importance. Eleven species are included in
the key. Most of them are widespread through Europe (Rognes 2004) and have been frequently reported from both real cases and
carrion experiments, and the necessity of their inclusion into the key cannot be questioned. There are: Calliphora vicina, C. vomitoria, Chrysomya albiceps, Phormia regina, Protophormia terraenovae, Lucilia caesar, L. illustris, L. sericata. The author has also decided to add three additional species to the key: Cynomya mortuorum, Chrysomya megacephala, and Lucilia ampullacea. Recently, the larvae of Cynomya mortuorum were recorded from human corpses at least twice (Stærkeby 2001; Benecke 2002). Smith (1986) points out this species as rather
a late newcomer in comparison to other blowflies, but recent research on succession shows that in spring conditions Cynomya mortuorum may be among the first colonizers of pig carrion (Szpila et al. 2008).
Scanning electron microscopy images of the first instars of Calliphora croceipalpis Jaennicke, 1876; Chrysomya chloropyga (Wiedemann, 1818); Chrysomya marginalis (Wiedemann, 1830); and Chrysomya putoria (Wiedemann, 1830) (Diptera: Calliphoridae) are presented for the first time, and the following morphological structures are documented: pseudocephalon, antenna, maxillary palpus, facial mask, labial lobe, thoracic and abdominal spinulation, spiracular field, posterior spiracles, and anal pads. Light microscopy photographs and line illustrations are provided for the cephaloskeleton in lateral and ventral views, and the "ectostomal sclerite" and "chitinized teeth" of the cephaloskeleton are recognized as integral parts of the mouthhooks. New diagnostic features of the cephaloskeleton and the spinulation of the abdominal segments are described. These results allow refinement, clarification, and correction of earlier descriptions, which are reviewed. The relative taxonomic importance of various morphological characters of the first instars of necrophagous blow flies is discussed, and details of the cephaloskeleton and the spinulation of the abdominal segments are highlighted as the characters most useful for species identification. Finally, a key for identifying first instars of common African carrion blow flies is provided.
Decomposition processes and insect succession were analysed on a total of eight pig (Sus scrofa) carcasses. The survey was carried out in four different periods between November 2007 and August 2008 and on two different substrates (meadow, concrete floor close to a building). The experiments were placed in a rural site in the surroundings of Jena, Thuringia. The duration and specificity of the different decomposition stages were dependent on season, weather and quality and quantity of insect colonization. Whereas the carrion maintained a bloated appearance even after 133 days in winter, it reached the dry stage within 8 days in the summer months. The type of substrate had few effects on the decaying process, but the insects were generally more abundant on the meadow. In total, more than 57 species of Diptera belonging to 17 families and 48 species of Coleoptera belonging to 14 families were identified. Dominant species belonged to the families of Calliphoridae (n = 11 spp.), Sarcophagidae (n = 8), Muscidae (n = 9), Piophilidae (n = 3), Silphidae (n = 6), Dermestidae (n = 3), Nitidulidae (n = 4), Cleridae (n = 3) and Histeridae (n = 2). Remarkably, the rather common Nicrophorus species were completely absent, whereas the usually rare Necrodes littoralis (L.) was present in larger numbers. No distinct coincidence between the occurrence of a single species and a certain decomposition stage could be confirmed. A main objective of the study is the establishment of a forensic entomological database for Central Europe, especially Thuringia.
Although the Cerrado is the second major Brazilian biome, few studies have been undertaken about its entomofauna. Blowflies have an important role in forensic entomology, helping in the determination of postmortem intervals. The main goal of this exploratory study was to identify and to catalog the blowfly species associated with a pig carcass. The study was conducted in a pasture in Brasília, Distrito Federal. A pig (Sus scrofa) was killed with a .22 caliber shot in the frontal region of the head. Adult blowflies were surveyed daily from June 1 through 30, 2004. A total of 14,910 adult calliphorids were collected, representing eight species: Chrysomya albiceps, C. megacephala, Cochliomyia macellaria, Chloroprocta idioidea, Hemilucilia semidiaphana, H. segmentaria, Lucilia cuprina, and L. eximia. C. albiceps was the most frequent species, amounting to 94.76% of the catch. Five decomposition stages were observed, and for calliphorids, the most attractive stage was the bloated one.
Various modification of the mixture of gum arabic with chloral hydrate can be used for mounting of small arthropods as well as some helminths. However, in diagnostic laboratories in Poland they seem to remain unknown. The authors of current work present examples of the Hoyer's medium application. The medium has been composed according to the initial, given by Hoyer, hundred-years-old recipe, which was the root of all other, later used recipes. Hoyer's medium is universal in action and very comfortable to use in slides for microscope investigation. It gives the immediate light-through effect, so it can be helpful in fast diagnostics. At the same time it allows to store the slides for a relatively long time e.g. with education purpose.
Larvae of 13 blowfly species from Colombia are described and an illustrated key for all them is presented. All larval instars of Calliphora nigribasis Macquart, Cochliomyia macellaria (Fabricius), Chrysomya albiceps (Wiedemann), Chrysomya megacephala (Fabricius), Hemilucilia segmentaria (Fabricius), Hemilucilia semidiaphana (Rondani), Lucilia eximia (Weidemann) are described, but the second and third instars of Compsomyiops verena (Walter), and only the third instar of Calliphora vicina Robineau-Desvoidy, Cochliomyia hominivorax (Coquerel), Lucilia peruviana Robineau-Desvoidy, Lucilia sericata (Meigen) and Sarconesiopsis magellanica (Le Guillou).
Morphology and duration of the post-embryonary development of Chrysomya megacephala (Diptera: Calliphoridae) in laboratory conditions. The morphology and developmental time of the post-embryonic stage of Chrysomya megacephala (Fabricius) reared in laboratory on a beef diet, is described and compared to those of other blow-flies species. The parameters analyzed were the duration of the larval stages and the morphology. Larvae were fixed and preserved at 2 hours intervals, from larval eclosion until the 50th hour of life, in order to determine the duration of each stage. The total period of larval development in C. megacephala was 98 hours and their mean length was 15.51 mm. This species presents a necrobiontophagous behavior and has sanitary and medico-legal importance, being used for estimating postmortem interval in criminal investigations.
The Namibian Calliphoridae fauna is reviewed. Seventy-five nominate species are recorded from the country, based on 7100 specimens housed in the Namibian National Insect Collection. Identification keys to all taxa are provided, together with notes on biology and new biological observations. An annotated checklist of Namibian species is presented, detailing previously published and other known Namibian records, and distributions elsewhere in Africa. Provisional distribution maps are plotted and discussed. Seasonal data, based on specimen label information, are presented in tabular form. The known immature stages of species are reviewed, together with notes on the forensic significance of relevant species. Two new species of the genus Hemigymnochaeta Corti and one new species of Isomyia Walker are described and illustrated, namely: Hemigymnochaeta maraisi Kurahashi and Kirk-Spriggs, sp. nov., Hemigymnochaeta trichaeta Kurahashi and Kirk-Spriggs, sp. nov., and Isomyia hauwangai Kurahashi and Kirk-Spriggs, sp. nov. Rhyncomya zumpti Peris, 1952 stat. rev., formerly synonymised with R. tristis Séguy, 1933, is here reinstated as a valid species. Thirty-four species are new to the Namibian list, namely: Bengalia spinifemorata Villeneuve, Chrysomya ?laxifrons Villeneuve, C. putoria (Wiedemann), Cosmina fuscipennis Robineau-Desvoidy, C. undulata Malloch, Fainia albitarsis (Macquart), Hemigymnochaeta bequaerti Curran, H. laticeps Zumpt, Isomyia darwini (Curran), I. deserti (Karsch), I. natalensis (Villeneuve), I. tristis (Bigot), Neocordylobia roubaudi Villeneuve, Perisiella anchora (Wiedemann), Phumosia nanoides Zumpt, Pseudorhyncomyia braunsi (Villeneuve), Rhyncomya bicolor (Macquart), R. cassotis (Walker), R. discrepans Villeneuve, R. hessei Zumpt, R. messoria Villeneuve, R. minutalis Villeneuve, R. io Peris, R. peraequa Villeneuve, R. soyauxi Karsch, R. trispina Villeneuve, R. tristis Séguy, R. zumpti Peris, Stegosoma vinculatum Loew, Stomorhina chapini Curran, S. rugosa (Bigot), Tricyclea claripennis Séguy, T. flavida (Malloch), and T. martini (Zumpt). Records of species from other southern African countries, based on 190 specimens housed in the Namibian National Insect Collection, are provided as an Appendix. Cosmina gracilis Curran and Rhyncomya dasyops Bezzi are new to Angola; Auchmeromyia bequaerti Roubaud and Bengalia cuthbertsoni Zumpt are new to Botswana; B. floccosa (van der Wulp), B. peuhi (Brauer and Bergenstamm) and Hemigymnochaeta unicolor (Bigot) are new to Zambia; and Cosmina gracilis Curran and Tricyclea martini (Zumpt) are new to Zimbabwe.
1. The relation of blowfly populations to season and to vegetational environment was studied over a number of years by serial trappings on the periphery of an aerodrome in a mixed-farming area near Carlisle. 2. Throughout the blowfly season the population consisted mainly of Calliphora erythrocephala, and this species was the first to appear and the last to disappear. The Lucilia caesar group formed a second major component in the summer months, representing up to 40% of the total. Within this group, L. illustris was slightly more frequent than L. caesar and L. ampullacea relatively scarce. Lucilia sericata and L. silvarum, Phormia terrae-novae and Cynomyia mortuorum were all present in low numbers only; Calliphora vomitoria occasionally represented up to 10% in the late season. Other species of Calliphorine blowflies were only rarely caught. Differences between species occurred in the sex-ratio of the trapped samples, and these varied between seasons and for different types of habitat. 3. A species-pattern similar to the above was obtained on reclaimed bogland and on upland sheep-grazing. 4. The general proportions of the major species-groups in the population were confirmed by tent-trapping of the ground without attractant bait. 5. Differences in total catch and in species-pattern between the five different vegetational habitats sampled are analysed in relation to season. 6. Calliphora vomitoria and Lucilia ampullacea apparently need dense cover such as wood. Calliphora erythrocephala was more numerous in the open near hedgerows than in wood, Lucilia caesar preferred tall-field-layer with some interrupted canopy, and L. illustris rather more open conditions. There is possibly a shift of some species of the caesar group population into more open conditions in the late part of the season. Relatively more males of Calliphora and Lucilia were caught by bait-traps in canopy habitats than elsewhere; this was not confirmed by the tent-trap catches. 7. The populations on different sections of the area in August, September and October were examined by intensive bait-trapping and by marking flies and releasing them again. 8. Calliphora erythrocephala was estimated at 50 to 200 flies per acre in August, 400 to 1000 in September, and 700 to 1000 in October. The caesar group was estimated as 40 to 70 per acre in August and September, and under 1 per acre (Lucilia caesar) in October. Lucilia sericata and Cynomyia mortuorum were estimated to be in very low numbers. 9. Bait-trap estimates of relative numbers showed significant differences in total blowfly population between the sections. The relative order of population-density of sections may change between tests, and sometimes within a few days during a single test. This latter effect is apparently due to the Calliphora erythrocephala component. 10. Bait-trap estimates of the population of ecologically uniform subsections of each section showed significant differences in population level and also differences in species-pattern. The relative level of population in different vegetational types varied between tests, and in the case of C. erythrocephala even within a few days. The apparent preferred habitats of the different species in the subsection tests were as found in the serial trap results, and the males of C. erythrocephala and the caesar group similarly were relatively more numerous in canopy cover. 11. The concept of arenas of activity as applied to blowflies is outlined in general terms. 12. Some limitations to the use of the release-recapture method of estimating numbers are discussed in relation to the results.
Keys and diagnostic descriptions are given for the eggs, three larval stages, and puparia of Megaselia scalaris (Loew), Piophila casei (L.), Muscina assimilis (Fallén), Muscina stabulans (Fallén), Chrysomya rufifacies (Macquart), Cochliomyia macellaria (F.), Phormia regina (Meigen), Calliphora vicina (Robineau-Desvoidy), Calliphora livida Hall, Phaenicia sericata (Meigen), Phaenicia coeruleiviridis (Macquart), and Lucilia illustris (Meigen). Some immature stages of the following are also included: Chrysomya chlorophyga putoria (Wiedemann), Calliphora peruviana (Robineau-Desvoidy), Phaenicia cuprina (Wiedemann), Phaenicia eximia (Wiedemann), and Phaenicia ibis (Shannon).
Lucilia cuprina and Lucilia sericata are probably the two most important sheep myiasis flies in the southern hemisphere. This paper describes and illustrates structural features that enable reliable specific identification of the first and second instars of these species of be made.
Differentiation of the third instar of forensically important fly species in Thailand was performed using light microscopy, based on their morphological criteria for fly identification. Four species of the family Calliphoridae [Chrysomya rufifacies (Macquart), Chrysomya megacephala (F.), Chrysomya nigripes Aubertin, and Lucilia cuprina (Wiedemann)] and two species of the family Muscidae [Musca domestica L. and Hydrotaea (=Ophyra) spinigera Stein] were examined in this study, with the features of the anterior spiracles, dorsal spines between the prothorax and mesothorax, and posterior spiracles being emphasized. The comparisons, presented herein, should be helpful for forensic practitioners to readily distinguish the third instars of fly species found associated with human cadavers, before their use for further forensic investigations.
In Europe, the blowfly genus Lucilia is represented in Forensic Entomology mainly by the species L. ampullacea, L. caesar, L. illustris and L. sericata. In the US, Lucilia silvarum is rarely recorded as a carrion breeding species but usually as a more or less exclusive parasite of frogs and toads. We present three forensic cases from different European countries reporting, for the first time, L. silvarum on human bodies that were found close to lakes, wetlands, or riversides. To use this species for post-mortem interval estimations, thermal development data is needed. The first step is accurate identification by morphological and molecular means. Therefore, we analysed a 611 bp part of the mitochondrial COI region for 23 specimens of L. silvarum from 9 different geographical regions, all of which give the same haplotype. Differences within the haplotype varied by up to 0.2%. Comparison between the haplotype found and those published on GenBank showed up to 1.2% variance. Moreover, we present an updated key for the morphological identification of the third larval instars of European Lucilia spp. of forensic importance, adding not only L. silvarum, but also L. cuprina which was recorded in Europe for the first time about 20 years ago.
Eggs of Calliphora augur (Fabricius), Calliphora stygia (Fabricius), Lucilia cuprina (Wiedemann) and Australophyra rostrata (Robineau-Desvoidy) are described and illustrated in detail by scanning electron micrographs.
First and second instar larvae of the above species along with those of Chrysomya rufifacies (Macquart), Chrysomya varipes (Macquart), Chrysomya saffranea (Bigot), Chrysomya nigripes Aubertin and first instar larvae of two species of sarcophagid (Trieholioproctia sp. and Parasarcophaga crassipalpis) are described, illustrated and compared with each other by means of keys. Particular attention is paid to the morphology of the pharyngeal skeleton and distribution of spines and, in the second instar larvae, to the morphology of the posterior spiracles and the number of processes on the anterior spiracles in addition to the characters mentioned previously.
1. An outdoor cage experiment was conducted to test the theory that localized interactions facilitate coexistence of species breeding in ephemeral habitats. The same amount of larval resource (50 g of liver) but divided into one, two, four, eight or sixteen pieces was placed into each of fifteen cages once a week from June to September for 4 years.
2. The cage populations were initially mixtures of thirteen species but after 4 years only Lucilia illustris (Meig.), Sarcophaga scoparia Pand. and S.aratrix Pand. remained. Lucilia illustris , by far the most abundant species in the field, predominated in every cage.
3. Sarcophaga went extinct in most cages, but they survived in five cages in which the larval resource was much divided, in agreement with the theoretical prediction.
4. Lucilia silvarum (Meig.) emerged about a week later than L.illustris in early summer; this phenological difference gave rise to a competitive asymmetry between the two species. This difference appears to explain why L.silvarum went extinct in the cages in 3 years, and it may explain why L.silvarum has consistently been less abundant than L.illustris in the natural community.
5. Other differences in phenology, voltinism and rate of development in the flies are described, and such differences are suggested to facilitate regional coexistence of many species in seasonal environments.
Abstract Scanning electron microscopy documentation of first instar Calliphora vicina Robineau-Desvoidy, Phormia regina (Meigen) and Lucilia illustris (Meigen) (Diptera: Calliphoridae) is presented for the first time, and the following morphological structures are documented: pseudocephalon; antenna; maxillary palpus; facial mask; labial lobe; thoracic and abdominal spinulation; spiracular field; posterior spiracles, and anal pad. Light microscopy documentation and illustrations are provided for the cephaloskeleton in lateral and ventral views. New diagnostic features are revealed in the configuration of the facial mask, cephaloskeleton and posterior spiracles. The first instar morphology of C. vicina, Ph. regina and L. illustris is discussed in the light of existing knowledge about early instars of blowflies.
To investigate the monophyly and phylogenetic relationships of the Calliphoridae (blowflies), 23 terminal monophyletic taxa within the Oestroidea and two outgroup taxa (Muscidae and Anthomyiidae) were scored for 45 adult and larval characters and subjected to analyses using parsimony. Search for trees of maximum total fit (using Pee-Wee) gave nine different trees for the possible values of the concavity constant, whereas search for minimum length trees under equal weights (using NONA and PAUP) gave two, different, trees. The preferred trees are those obtained from the maximum-fit searches since they are derived from a method that lowers the weight of characters showing within-terminal as well as within-tree homoplasy. The family Calliphoridae in various commonly used senses (excluding Oestridae s.lat.; including the Rhiniinae; excluding or including the Rhinoporidae; excluding or includingMystacinobia) does not appear to be a monophyletic group. It appears as such only in trees having a much lower total fit than the optimal trees (maximum fit approach) or only in a very small fraction of the numerous trees five or nine steps longer than the minimum length trees. The assemblage (Rhiniinae+Toxotarsinae+Chrysomyinae), sometimes assumed to be a natural taxon on account of common possession of a row of setae on the upper surface of the stem-vein, should likewise be rejected. A clade (Ameniini+Euphumosia+Phumosiinae+Mesembrinellinae+Parameniini+Catapicephala) is possibly Gondwanian. A clade (Toxotarsinae+Chrysomyinae+Calliphorinae+Luciliinae+Melanomyinae) contains all the sarcosaprophagous “blowflies”. The group (Tachinidae+Sarcophagidae) is possibly the monophyletic sister group of the Rhiniinae.Mystacinobiaemerges from the analyses as the sister group of all other Oestroidea, and the family Axiniidae as the sister group of all the other non-mystacinobiine oestroids. AlthoughMystacinobiais a guano-feeder in all stages, the biology of the other basal oestroid taxa indicates that a reproductive association with living or dead invertebrates is the primitive oestroid pattern. Fossils having a bearing upon the estimation of the age of various oestroid clades are discussed.
Scanning electron microscopy was used to determine whether first instar larvae were capable of physically damaging sheep skin and thus initiating blowfly strike without predisposing conditions. In addition, damage at the lesion site was monitored throughout the infection period. It was found that first instar larvae do not possess the mouth hooks seen in second and third instars. They do however have sets of spines within the oral cavity which, with the spines on the creeping welts, may be able to abrade the outermost skin cell layers. Damage to the epidermis was observed within the first 8 h after larval implantation on sheep. The dermis was exposed within the first 24 h and after this time the normal skin architecture was rapidly destroyed. At 72 h areas of wool were missing and the collagen matrix of the dermis was clearly visible. Predisposing conditions such as fleece rot or wounding were not necessary for strike initiation. It is suggested that a combination of physical irritation and abrasion together with enzymes released by larvae enable them to initiate formation of the strike lesion. Thus, the only mandatory predisposing condition for fly strike may be wetted wool and skin.
The insect fauna of pig carcasses was monitored in different seasons and forests of Western Poland (Central Europe). The composition of carrion fauna and selected features of residency in carrion in adults and larvae of particular taxa were analysed. A total of 131 adult and 36 larval necrophilous taxa were collected. Only 51 adult species and 24 larval taxa were minimally abundant (≥10 specimens) at least on one carcass. As for the composition of carrion fauna, there were large differences between seasons, but no important differences between forest types.
The Calliphoridae include some of the most economically significant myiasis-causing flies in the world - blowflies and screwworm flies - with many being notorious for their parasitism of livestock. However, despite more than 50 years of research, key taxonomic relationships within the family remain unresolved. This study utilizes nucleotide sequence data from the protein-coding genes COX1 (mitochondrial) and EF1α (nuclear), and the 28S rRNA (nuclear) gene, from 57 blowfly taxa to improve resolution of key evolutionary relationships within the family Calliphoridae. Bayesian phylogenetic inference was carried out for each single-gene data set, demonstrating significant topological difference between the three gene trees. Nevertheless, all gene trees supported a Calliphorinae-Luciliinae subfamily sister-lineage, with respect to Chrysomyinae. In addition, this study also elucidates the taxonomic and evolutionary status of several less well-studied groups, including the genus Bengalia (either within Calliphoridae or as a separate sister-family), genus Onesia (as a sister-genera to, or sub-genera within, Calliphora), genus Dyscritomyia and Lucilia bufonivora, a specialised parasite of frogs and toads. The occurrence of cross-species hybridisation within Calliphoridae is also further explored, focusing on the two economically significant species Lucilia cuprina and Lucilia sericata. In summary, this study represents the most comprehensive molecular phylogenetic analysis of family Calliphoridae undertaken to date.
Seasonal and habitat calliphorid abundance and distribution were examined weekly for two years (2001-2003) in Santa Clara County, California, using sentinel traps baited with bovine liver. Of the 34,389 flies examined in three defined habitats (rural, urban, and riparian), 38% of the total catch represented Compsomyiops callipes (Bigot) and 23% represented Phormia regina (Meigen). Other flies collected in this survey included Calliphora vomitoria (Linnaeus), Calliphora latifrons (Hough), Lucilia sericata (Meigen), Lucilia cuprina (Wiedemann), and Lucilia mexicana (Macquart), which is a new record for the area. Multivariate MANOVA and ANOVA (P ≤ 0.05) analysis indicate significant seasonal habitat preference for all fly species examined. This information may be used to identify potentially forensically impo rtant fly species within Santa Clara County, California.
This article presents morphological descriptions of larvae and adults of Hypopygiopsis tumrasvini Kurahashi (Diptera: Calliphoridae), a blow fly species that may be of forensic importance. Both second and third instar larvae are illustrated, with emphasis placed on important characteristic features used for larval identification, such as the cephalopharyngeal skeleton, anterior and posterior spiracles, and dorsal spines between the prothorax and mesothorax. Eight to 11 papillae were found on each anterior spiracle arranged in a single row. The dorsal spines between the prothorax and mesothorax were arranged in sets of posteriorly projecting acuminate spines with darkly pigmented tips. The posterior spiracles each bear three prominent and separated long, slender spiracular slits encircled by a dark, thick peritreme that is complete ventromedially around a button. Prominent inner projection of peritreme is seen between the middle and lower spiracular slits. A previously published key for differentiating third instar larvae of flies of possible forensic importance in Thailand is updated to include this additional species. Some characteristic features of males and females of the species are also provided.
Hypopygiopsis violacea, a species of fly of forensic importance, was recovered from a corpse and described for the first time. The morphological structures of the second and third instar larvae of four specimens were examined using light microscope. Observations were focused on three main morphological characters: cephalopharyngeal skeleton, anterior and posterior spiracles. Cephalopharyngeal skeleton of second instar larva is darkly pigmented and without accessory sclerite below the mouth hook. The anterior spiracles of second and third instar larvae have 8-9 papillae each, arranged in a single row. The posterior spiracle of second instar larva has two spiracular slits with no thickening of peritreme. This differentiates it from the third instar, whereby the latter has three slits for each posterior spiracle. Cephalopharyngeal skeleton of third instar larva is heavily pigmented. An accessory sclerite is found below the hook part of third instar larva but is absent in second instar. Peritreme of the posterior spiracle of third instar larva is thick almost complete encircling a button. The intersegmental spines of the cuticular surface are dome-shaped and unicuspid. Third instar larva of this species is large with size approximately 15 mm long. These findings provide important identification features of immature stages of Hy. violacea which could be useful in forensic entomology.
Lucilia cuprina and Hemipyrellia ligurriens are forensically important blow flies in many countries. The immature stage, in particular the third instar, of both species are generally similar in morphological appearance. In this study, the use of three main characteristics, the posterior spiracle, the dorsal spines between the first and second thoracic segments, and the cephalopharyngeal skeleton, allows for identification of both fly species, with the number of papillae on anterior spiracle being a supportive characteristic in some cases. Comparison of these characteristics is illustrated in detail.
At the end of summer and beginning of fall 2005 insect succession and pig carrion decomposition were studied in pine-oak forest, hornbeam-oak forest, and alder forest in Western Poland (Central Europe). Five stages of decomposition were recognized: fresh stage, bloated stage, active decay stage, advanced decay stage, and remains stage. The highest rate of decomposition was observed in the alder forest and the lowest in the pine-oak forest. A similar composition of carrion entomofauna was found in each forest. Numerous adults of Lucilia caesar, Phormia regina and Calliphora vomitoria (Calliphoridae), Hydrotaea spp. (Muscidae), Stearibia nigriceps and Parapiophila vulgaris (Piophilidae), Necrodes littoralis and Thanatophilus rugosus (Silphidae), Creophilus maxillosus, Omalium rivulare, Oxypoda acuminata and Philonthus spp. (Staphylinidae) as well as larvae of L. caesar, P. regina, C. vomitoria, Fannidae (Diptera) and N. littoralis, C. maxillosus, Philonthus (Coleoptera) were collected. There were no differences between forests in the sequence of insect occurrence on carrion. However, differences between forests in occurrence time and activity period of some taxa were found. Implications of these results for forensic entomology are discussed.
The reliable morphological identification of carrion-breeding blowflies is important ecologically, as well as for medical, veterinary and forensic reasons. To date, no comprehensive key has been available to make this possible for workers in southern Australia. An illustrated key is presented to the adults of all species of blowflies (Diptera: Calliphoridae) south of 30 degrees S known or suspected to breed in carrion (species exclusive to Queensland and/or the Northern Territory are excluded).
Complementary nuclear (28S rRNA) and mitochondrial (COI + II) gene markers were sequenced from the blowflies, Lucilia cuprina and Lucilia sericata, from Europe, Africa, North America, Australasia and Hawaii. Populations of the two species were phylogenetically distinct at both genes, with one exception. Hawaiian L. cuprina possessed typical L. cuprina-type rRNA, but had L. sericata-type mitochondrial (COI + II) sequences. An explanation for this pattern is that Hawaiian flies are hybrids and comparison of observed levels of sequence divergence to possible introduction events, e.g. Polynesian colonization, suggests that Hawaiian L. cuprina may be evolving rapidly. Moreover, the monophyly of these flies also suggests that the L. sericata mtDNA haplotype was apparently fixed in Hawaiian L. cuprina by lineage sorting, indicating a population bottleneck in the evolutionary history of these island flies.
The larval morphology of Chrysomya megacephala (Fabricius) is presented using scanning electron microscopy (SEM). Extreme similarity of this species to Chrysomya rufifacies (Macquart), a species usually found concurrently inhabiting decomposing human corpses in Thailand, is seen only in the first-instar larvae. The relative thickness of the branches of the posterior spiracular hairs in these species could be used to differentiate them in this developmental stage. In contrast, the "hairy" appearance of C. rufifacies allows second- and third-instar larvae to be easily distinguished. Results of this study should help in future endeavors to differentiate C. megacephala from other larvae found in decomposing human corpses in Thailand.
The surface ultrastructure of all larval instars of Chrysomya rufifacies (Macquart) is described by means of scanning electron microscopy (SEM). Morphological changes were greatest from the first to the second instar, but less from the second to the third instar. Most of these changes involved the structure of the anterior spiracle, posterior spiracle, integument of the body, and mouthhooks. Modification of the mouthhooks, especially in the third instar, are helpful in explaining the ferocious feeding ability of the older maggots. The common name of "hairy-maggot" for C. rufifacies is only appropriate for the second and third instars because of their elongated tubercles along the body, whereas this name is not descriptive of the first instar that lack tubercles.