Geographic origin affects larval competitive ability in European populations of the blow fly, Lucilia sericata

The blow fly, Lucilia sericata (Meigen) (Diptera: Calliphoridae), is found throughout Europe. In northern areas, in addition to breeding in carrion, it can also commonly act as primary facultative ectoparasite of sheep, resulting in animal suffering and production losses. However, in the south of Europe, L. sericata is not usually implicated in myiasis and is found almost entirely in carrion. The causes of this behavioural difference are unclear. To begin to address the possible reasons for this difference, the present study examined the effects of competition on the survival and development of L. sericata populations derived from southern Spain, England, and a hybrid derived from a cross between Spanish females and UK males. Larvae were reared at a range of different initial densities and the mortality, development rate, and size of the resultant adults were measured. Mortality increased significantly with density, but the Spanish population had a significantly higher mortality rate than the UK population at all densities. Similarly, adult size declined with increased larval density, but individuals from the Spanish population were consistently larger than those from the UK population at all densities. The mortality and size of the hybrids was consistently intermediate between the Spanish and UK populations. No consistent effects on development rate were observed. This study demonstrates that insects from northwestern and southern European population have qualitatively different life history traits and competitive abilities and may contribute towards explaining the different behaviour of populations of this species in different part of its distribution range.     

Observations on myiasis by the calliphorid,Bufolucilia silvarum,in the eastern American toad (Bufo americanus americanus) from southeastern Wisconsin

Larvae of certain species of blowflies (Calliphoridae) cause myiasis in amphibians which may result in significant mortality, yet there are few reports from North America. In this study, we observed primary myiasis in a population of juvenile eastern American toads (Bufo americanus americanus) collected during May-July 1998 from southeastern Wisconsin (USA). Nine (6%) of 140 toads were infected by the green blow fly (Bufolucilia silvarum) with a mean intensity of 10.5 +/- 7.2 (range = 1-24). Weekly parasite prevalence and mean intensity remained low, ranging from 0-20% and 2 +/- 1.4 to 14 +/- 6, respectively. We found: 1) flies lay eggs on healthy toads, 2) eggs hatch with first instar maggots penetrating under the skin, 3) maggots develop to mature third instars within 5-7 days, 4) maggots leave the host and form pupa within 8-11 days of hatching, and 5) maggots pupate within 7-9 days at room temperature. All infected toads died within 1-2 wk as a result of the infection. The low prevalence observed in this study and other reports of this species from mammalian and bird carcasses indicated that B. silvarum is probably a facultative parasite of toads and other amphibians in the United States. This is the first report of B. silvarum causing myiasis in Wisconsin amphibians and the first report in eastern American toads in the United States.     

Larval Distribution and Behavior of Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) Relative to Other Species on Florida Black Bear (Carnivora: Ursidae) Decomposing Carcasses

Larval interactions of dipteran species, blow flies in particular, were observed and documented daily over time and location on five black bear carcasses in Gainesville, FL, USA, from June 2002 – September 2004. Cochliomyia macellaria (Fabricius) or Chrysomya megacephala (Fabricius) larvae were collected first, after which Chrysomya rufifacies (Macquart) oviposited on the carcasses in multiple locations (i.e., neck, anus, and exposed flesh) not inhabited already by the other blow fly larvae. Within the first week of decomposition, C. rufifacies larvae grew to ≥12 mm, filling the carcasses with thousands of larvae and replacing the other calliphorid larvae either through successful food source competition or by predation. As a result, C. macellaria and C. megacephala were not collected past their third instar feeding stage. The blow fly species, C. megacephala, C. macellaria, Lucilia caeruleiviridis (Macquart), Phormia regina (Meigen), Lucilia sericata (Meigen), and C. rufifacies, completed two developmental cycles in the 88.5-kg carcass. This phenomenon might serve to complicate or prevent the calculation of an accurate postmortem interval.     

Spatial and temporal habitat partitioning by calliphorid blowflies

Calliphorid blowflies perform an essential ecosystem service in the consumption, recycling and dispersion of carrion nutrients and are considered amongst the most important functional groups in an ecosystem. Some species are of economic importance as facultative agents of livestock myiasis. The interspecific ecological differences that facilitate coexistence within the blowfly community are not fully understood. The aim of this work was to quantify differences in habitat use by calliphorid species. Thirty traps were distributed among three habitats at two sites in southwest England for collections made during March–August 2016. A total of 17 246 specimens were caught, of which 2427 were Lucilia sericata, 51 Lucilia richardsi, 6580 Lucilia caesar, 307 Lucilia ampullacea, 4881 Calliphora vicina and 2959 Calliphora vomitoria (all: Diptera: Calliphoridae). Lucilia sericata was the dominant species in open habitats, whereas L. caesar was the most abundant species in shaded habitats. Calliphora specimens were more abundant in the cooler months. These findings suggest that Calliphora and Lucilia species show strong temporal segregation mediated by temperature, and that species of the genus Lucilia show differences in the use of habitats that are likely to be driven by differences in humidity tolerance and light intensity. These factors in combination result in effective niche partitioning.     

Cloning,recombinant expression and inhibitor profiles of dihydrofolate reductase from the Australian sheep blow fly,Lucilia cuprina

While dihydrofolate reductase (DHFR) is an important drug target in mammals, bacteria and protozoa, no inhibitors of this enzyme have been developed as commercial insecticides. We therefore examined the potential of this enzyme as a drug target in an important ectoparasite of livestock, the Australian sheep blow fly, Lucilia cuprina (Diptera: Calliphoridae) (Wiedemann). The non‐specific DHFR inhibitors aminopterin and methotrexate significantly inhibited the growth of L. cuprina larvae, with IC₅₀ values at µg levels. Trimethoprim and pyrimethamine were 5–30‐fold less active. Relative IC₅₀ values for the inhibition of recombinant L. cuprina DHFR by various inhibitors were in accordance with their relative effects on larval growth. The active‐site amino acid residues of L. cuprina DHFR differed by between 34% and 50% when compared with two mammalian species, as well as two bacteria and two protozoa. There were significant charge and size differences in specific residues between the blow fly and human DHFR enzymes, notably the L. cuprina Asn21, Lys31 and Lys63 residues. This study provides bioassay evidence to highlight the potential of blow fly DHFR as an insecticide target, and describes differences in active site residues between blow flies and other organisms which could be exploited in the design of blow fly control chemicals.     

Myiasis by Lucilia silvarum (Calliphoridae) in amphibian species in boreal Alberta, Canada

We report myiasis by Lucilia silvarum with an overall prevalence of 0.9% in amphibian populations in boreal Alberta. In the period 1998--1999, we documented L. silvarum infestations in wild populations of wood frog (Rana sylvatica), boreal chorus frog (Pseudacris maculata), boreal toad (Bufo boreas boreas), and Canadian toad (B. hemiophrys). We believe this is the first record of this parasite from boreal and Canadian toads. Almost all previous records of L. silvarum parasitism in North America indicate that myiasis is fatal to an anuran host. Here, we provide the first record of adult individuals from 2 species (wood frog and boreal toad) surviving infestations. Although we actively captured and examined amphibians in Alberta from 1996 to 2006, we only found parasitism in 1998 and 1999. This is the most northerly record of anuran infestations by this parasite.     

Traumatic Myiasis Caused by an Association of Sarcophaga tibialis (Diptera: Sarcophagidae) and Lucilia sericata (Diptera: Calliphoridae) in a Domestic Cat in Italy

We describe here a rare case of traumatic myiasis occurred in August 2014, caused by an association of 2 Diptera species, Sarcophaga tibialis Macquart (Diptera: Sarcophagidae) and Lucilia sericata (Meigen) (Diptera: Calliphoridae), in a domestic cat in northern Italy. Species identification was based on adult male morphology. The present case is the first report of S. tibialis as an agent of myiasis in Italy, and also the first ever report of myiasis caused by an association of S. tibialis and L. sericata. The cat developed an extensive traumatic myiasis in a large wound on the rump, which was treated pharmacologically and surgically. The biology, ecology, and distribution of S. tibialis and L. sericata are also discussed. A literature review is provided on cases of myiasis caused by S. tibialis, and cases of myiasis by L. sericata involving cats worldwide and humans and animals in Italy.     

Bimodal cue complex signifies suitable oviposition sites to gravid females of the common green bottle fly

Gravid females of the common green bottle fly, Lucilia sericata Meigen (Diptera: Calliphoridae), readily locate recently deceased vertebrates as oviposition sites, particularly when these animals have been injured. We investigated semiochemical and visual cues that mediate attraction of gravid females to fresh rat carrion. Female flies were more strongly attracted to incised rat carrion than to intact carrion. They were also attracted to Porapak Q headspace volatile (HSV) extract of incised rat carrion. Analyzing aliquots of Porapak Q HSV extract by gas chromatographic‐electroantennographic detection revealed nine components [phenol, para‐ and/or meta‐cresol (could not be separated), guaiacol, dimethyl trisulfide (DMTS), phenylacetaldehyde, (E)‐2‐octenal, nonanal, and tetramethyl pyrazine] that consistently elicited responses from blow fly antennae. In laboratory experiments, a synthetic blend of these nine components was as attractive to gravid females as Porapak Q HSV extract, but blend attractiveness was due entirely to DMTS. In both laboratory and field experiments, increasing doses of DMTS attracted increasingly more flies. Coupled with DMTS, carrion‐type color cues (dark red, black) were more effective than bright color cues (white, yellow) in attracting flies. In field experiments, dark traps baited with DMTS captured a total of 214 calliphorid flies (200 L. sericata, 10 Lucilia illustris Meigen, three Calliphora vicina Robineau‐Desvoidy, one Calliphora vomitoria L.), all of which were gravid females. These results support the conclusion that DMTS and dark color represent a bimodal cue complex that signifies suitable oviposition sites to gravid calliphorid females, particularly L. sericata.     

Observations on myiasis by the calliphorids, Bufolucilia silvarum and Bufolucilia elongata, in wood frogs, Rana sylvatica, from southeastern Wisconsin

Larvae of certain species of blowflies (Calliphoridae) can cause myiasis in frogs and toads, but there are few reports from North American amphibians. Of these, most are from toads (bufonids). In this study, we observe primary myiasis in a population of juvenile wood frogs, Rana sylvatica, collected on 22-23 August 2003, from southeastern Wisconsin and compare our observations with previous studies on myiasis from toads. Two (5%) of 39 frogs were infected by the blow fly Bufolucilia silvarum, with an intensity of 28 and 31, whereas 1 (2.5%) of 39 frogs was infected by the blow fly Bufolucilia elongata with an intensity of 14. We found that (1) B. silvarum lay eggs on healthy wood frogs, (2) eggs hatch, with first-instar maggots penetrating under the skin, (3) maggots develop to mature third instars within 13-16 hr of egg hatching, (4) maggots kill the host within 7-47 hr of egg hatching, and (5) maggots consume the entire frog carcass reducing it to bones within 42-59 hr of egg hatching. Our observations on the time of death and how quickly carcasses of wood frogs were consumed by these maggots compared with previous studies on toads suggest that finding infected juvenile wood frogs may be uncommon. Therefore, myiasis by these flies on wood frogs and other small terrestrial anurans may be a phenomenon that is much more common than is currently observed. This is the first report of B. silvarum and B. elongata causing myiasis in wood frogs.     

Complex competitive interactions in four species of Lucilia blowflies

Abstract.

• 1 The effects of intraspecific and interspecific larval competition on larval survival, adult size, adult longevity and fecundity were quantified in four species of coexisting Lucilia blowflies: illustris, silvarum, sericata and caesar.
• 2 There was a general negative effect of increasing density on larval and adult survival, size and fecundity. Additionally, complex species-specific and frequency-dependent responses were identified, which were not expected in these biologically and morphologically closely similar species.
• 3 Lucilia illustris, the numerically dominant species in the natural community, was a superior competitor to L.silvarum at intermediate densities but an inferior competitor at high density. Such nonlinear responses may be related to differences in the life histories and larval behaviour of the species (bigger eggs and more contest-type outcome of competition in L.silvarum).
• 4 We parameterized a model of interspecific competition on a subdivided resource in an attempt to reconcile the conflicting results on larval competitive abilities and the abundances of the species in the field. Using laboratory and field-estimated parameter values the model predicted coexistence of L.illustris and L.silvarum and the observed numerical dominance of the former species. The average densities of flies in the field are limited to relatively low levels, apparently preventing L.silvarum (the superior competitor at high density) from dominating and excluding L. illustris.

     

Mortality in a population of Bufo bufo resulting from the fly Lucilia bufonivora

Summary In the summers of 1972 and 1973, in an area of inland dunes in the Netherlands, and estimation was made of the percentage of amphibians that was killed by infections caused by Lucilia bufonivora. It appeared that only Bufo bufo was preyed on. In the two years of the investigation an average of 8% of the toads found was infected. The flies selected their prey according to their size. Predation took place in every habitat of the study area.     

Blow fly Trypocalliphora braueri parasitism on Meadow Pipit and Bluethroat nestlings in Central Europe

The larvae of blow fly (Trypocalliphora braueri) are known to cause a subcutaneous myiasis in bird nestlings mainly in Nearctic species, while only a few records are available from the Palaearctic. Here, we report on infestations of this insect ectoparasite in two passerine species, Meadow Pipit (Anthus pratensis) and Bluethroat (Luscinia svecica), breeding in the Central European mountain range Krkonoše. The infestation of T. braueri parasite was relatively low in studied bird species (3.0% of infested nests in Meadow Pipit and 4.2% in Bluethroat) but varied strongly between years (0–33% of infested nests/year/species). The presence of the parasitic blow fly larvae was apparently controlled by temperature; they were found only during warm summers. The combination of parasite infection and heavy rainfalls in a critical period significantly reduced nestling survival.     

Identification of forensically important blow fly species (Diptera: Calliphoridae) in China by mitochondrial cytochrome oxidase I gene differentiation

Abstract Unambiguous and rapid sarcosaphagous insect species identification is an essential requirement for forensic investigations. Although some insect species are difficult to classify morphologically, they can be effectively identified using molecular methods based on similarity with abundant authenticated reference DNA sequences in local databases. However, local databases are still relatively incomplete in China because of the large land area with distinct regional conditions. In this study, 75 forensically important blow flies were collected from 23 locations in 16 Chinese provinces, and a 278‐bp segment of the cytochrome oxidase subunit I gene of all specimens was successfully sequenced. Phylogenetic analysis of the sequenced segments showed that all Calliphorid specimens were properly assigned into nine species with relatively strong supporting values, thus indicating that the 278‐bp cytochrome oxidase subunit one region is suitable for identification of Calliphorid species. The clear difference between intraspecific threshold and interspecific divergence confirmed the potential of this region for Calliphorid species identification, especially for distinguishing between morphologically similar species. Intraspecific geographic variations were observed in Lucilia sericata (Meigen, 1826) and Lucilia caesar (Linnaeus, 1758).     

Influence of parasitism and soil compaction on pupation of the green bottle fly,Lucilia sericata

The influence of parasitoids and soil compaction on pupation behavior of blow flies was examined in a host–parasitoid system involving Lucilia sericata (Meigen) (Diptera: Calliphoridae) and Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae). Larvae of L. sericata were introduced to containers with soil of different compaction levels, with or without parasitoids. Although females of N. vitripennis did not significantly affect the pupation depth of L. sericata, they increased the rate of pupal development by 15.0–23.7 h at 28.4 ± 1.2 °C, and increased the clumping of puparia. Pupation depth of L. sericata was negatively related to soil compaction; mean depth of pupation was 4.4 cm in uncompacted soil and 0.5 cm in high‐compaction soil. In high‐compaction soil, pupal development increased by 10.5–18.8 h at 25.2 ± 0.3 °C, and puparia were clumped. These results provide a framework for locating puparia in forensic investigations and releasing appropriate parasitoids for biological control of blow flies.     

Synomones in necrophagous larvae of the blow flies Lucilia sericata and Calliphora vomitoria

Chemical signals are widespread in insects, but those resulting in interspecific communication (i.e., synomones) remain understudied. Here, we analysed chemicals left on substrates by two species of blow fly larvae, Lucilia sericata (Meigen) and Calliphora vomitoria (Linneaus) (Diptera: Calliphoridae), which can aggregate together on carrion. Using solid-phase microextraction and dynamic headspace analysis, we identified six compounds common to both species: the decanoic, tetradecanoic, pentadecanoic, hexadecanoic and octadecanoic acids, and the 2-ethylhexyl salicylate. We then tested the behavioural effects of the decanoic and pentadecanoic acids using binary-choice experiments, along with the (Z)-9-tricosene, a pheromone found in many arthropods. The time spent by a larva and its average crawling speed were measured in two sides of an arena, where only one contained a compound at 0.25 or 25 μg/μl. No effect was observed when testing the decanoic acid. The pentadecanoic acid only reduced the speed of C. vomitoria larvae at 25 μg/μl. Finally, L. sericata larvae spent less time in the side containing the (Z)-9-tricosene at 0.25 μg/μl, whereas C. vomitoria spent more time and crawled faster in this side at 25 μg/μl. Although these results did not directly evidence synomones, they suggest that the (Z)-9-tricosene could regulate larval aggregations on carrion.     

Successional patterns of the insect fauna on a pig carcass in southern Italy and the role of Crematogaster scutellaris (Hymenoptera,Formicidae) as a carrion invader

The sarcosaprophagous fauna plays a key role in organic matter decomposition. Moreover, the biological, ecological and behavioral specificities of the taxa are useful to reconstruct the decay history of a corpse or carcass, often back to the lethal event. Here we report the seasonal succession of the insect fauna on a pig carcass exposed in a rural area in Calabria (southern Italy) during summer 2007 and 2008. The aim is to identify and qualitatively assess the major taxa of forensic importance in this region. The principal fly invaders were Lucilia caesar (L.), L. sericata (Meigen, 1826), Chrysomya albiceps (Wiedemann, 1819), Sarcophaga (Meigen, 1826) spp. and Amobia (Robineau‐Desvoidy, 1830) spp., Musca domestica (L.) and Muscina stabulans (Fallen, 1817). The primary beetle species collected in summer belonged to Dermestidae, Dermestes maculatus (De Geer, 1774) and Cleridae, Necrobia rufipes (De Geer, 1775). This paper also examined the ecological role of ants in the insect succession and describes the evidence of skin injuries directly inflicted by the acrobat ant Crematogaster (Acrocoelia) scutellaris (Olivier, 1791) (Hymenoptera Formicidae) while feeding on pig carrion. Ants belonging to two other species were also collected: Camponotus aethiops (Latreille, 1798) and Tetramorium semilaeve (André, 1881). Ants can invade carcasses and corpses directly, disrupting blowfly egg laying or preying on their larvae. Our data on the carrion faunal composition and role of ants as invaders should be useful for further forensic cases in Calabria (southern Italy). This is among the few reports of ants as forensically relevant species.     

Amphibian road mortality in Europe: a meta-analysis with new data from Poland

While the increasing vehicular traffic is widely suspected to play a role in the worldwide amphibian population decline, the research of amphibian road mortality is scarce, fragmented, fraught with methodological problems, and largely inconclusive. As the first attempt at a synthesis, we analyzed all available data on amphibian mortality in Europe and combined them with four previously unpublished surveys conducted by us. Based on our recalculation of road-kill counts in terms of species-specific road-kill recordability, we conclude that, in lowland Central Europe, the common toads, Bufo bufo, are the most common victims of vehicular traffic in suburban landscapes, while the common frogs, Rana temporaria and Triturus newts, prevail in rural landscapes. The green frogs also tend to be more frequent in rural areas. Common tree frogs, Hyla arborea, are unexpectedly rare in the road-kill record despite their terrestrial and migratory habits. In consideration of problems with obtaining accurate amphibian population estimates, we further propose the road kills-to-spawners ratio (R/S) as a working measure of the impact of road mortality on a local population. While the R/S ratio may not reflect the losses to an entire local amphibian population, it is free of the errors of whole-population estimates, which are notoriously difficult for amphibians. When corrected for species-specific road-kill recordability, most results suggest that the impact of roads on newts may have been underestimated and that the impact on common frog populations may be higher than on those of common toads. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Communicated by H. Kierdorf     

Genetic Approaches for Studying Myiasis-causing Flies: Molecular Markers and Mitochondrial Genomics

“Myiasis-causing flies” is a generic term that includes species from numerous dipteran families, mainly Calliphoridae and Oestridae, of which blowflies, screwworm flies and botflies are among the most important. This group of flies is characterized by the ability of their larvae to develop in animal flesh. When the host is a live vertebrate, such parasitism by dipterous larvae is known as primary myiasis. Myiasis-causing flies can be classified as saprophagous (free-living species), facultative or obligate parasites. Many of these flies are of great medical and veterinary importance in Brazil because of their role as key livestock insect-pests and vectors of pathogens, in addition to being considered important legal evidence in forensic entomology. The characterization of myiasis-causing flies using molecular markers to study mtDNA (by RFLP) and nuclear DNA (by RAPD and microsatellite) has been used to identify the evolutionary mechanisms responsible for specific patterns of genetic variability. These approaches have been successfully used to analyze the population structures of the New World screwworm fly Cochliomyia hominivorax and the botfly Dermatobia hominis. In this review, various aspects of the organization, evolution and potential applications of the mitochondrial genome of myiasis-causing flies in Brazil, and the analysis of nuclear markers in genetic studies of populations, are discussed.