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The fauna of blowflies (Calliphoridae and Mesembrinellidae) in three localities of primary Amazon forest coverage in the Amazonas-Negro interfluvial region was assessed. A total of 5066 blowflies were collected, with Chloroprocta idiodea being the most abundant species (66.3%). A difference in species richness between the localities ZF2 and Novo Airão was observed. Comparison among sampled sites revealed no considerable variation in fauna composition, except for the species Eumesembrinella benoisti (Séguy 1925) and Hemilucilia sp., whose occurrence was observed only in a single locality. Apparently, Amazon rivers are not efficient geographical barriers to influence the current composition of necrophagous blowfly assemblages. Also, most of the blowfly species did not show a noticeable specificity for any specific forest among the interfluvial areas of the ombrophilous forest. Finally, an updated checklist of necrophagous blowfly species of the Amazonas state in Brazil is presented. Keywords: Blowfly assemblage, Calliphoridae, Diversity, Forensic entomology, Mesembrinellidae

Calyptrate flies include about 22,000 extant species currently classified into Hippoboscoidea (tsetse, louse, and bat flies), the muscoid grade (house flies and relatives) and the Oestroidea (blow flies, bot flies, flesh flies, and relatives). Calyptrates are abundant in nearly all terrestrial ecosystems, often playing key roles as decomposers, parasites, parasitoids, vectors of pathogens, and pollinators. For oestroids, the most diverse group within calyptrates, definitive fossils have been lacking. The first unambiguous fossil of Oestroidea is described based on a specimen discovered in amber from the Dominican Republic. The specimen was identified through digital dissection by CT scans, which provided morphological data for a cladistic analysis of its phylogenetic position among extant oestroids. The few known calyptrate fossils were used as calibration points for a molecular phylogeny (16S, 28S, CAD) to estimate the timing of major diversification events among the Oestroidea. Results indicate that: (a) the fossil belongs to the family Mesembrinellidae, and it is identified and described as Mesembrinella caenozoica sp. nov.; (b) the mesembrinellids form a sister clade to the Australian endemic Ulurumyia macalpinei (Ulurumyiidae) (McAlpine’s fly), which in turn is sister to all remaining oestroids; (c) the most recent common ancestor of extant Calyptratae lived just before the K–Pg boundary (ca. 70 mya); and (d) the radiation of oestroids began in the Eocene (ca. 50 mya), with the origin of the family Mesembrinellidae dated at ca. 40 mya. These results provide new insight into the timing and rate of oestroid diversification and highlight the rapid radiation of some of the most diverse and ecologically important families of flies. ZooBank accession number–urn:lsid:zoobank.org:pub:0DC5170B-1D16-407A-889E-56EED3FE3627.

Paralucilia Brauer & Bergenstamm, 1891 (Diptera, Oestroidea, Calliphoridae) is a small genus of blowflies restricted to the Neotropical region, which is commonly reported on decaying corpses and vertebrate carcasses. The number of species currently assigned to this genus and their denominations are contentious, with either three or five species recognized by different authors. This taxonomic instability results in a lack of consensus in species determination, making it impossible to compare results from different studies as well as to elaborate confident taxonomic keys. In order to solve some of the most commonly reported taxonomic conflicts within this genus, to the best of our knowledge, this study presents the first phylogenetic hypothesis for the relationships among Paralucilia species, based on molecular analysis of the COI, ITS2, 28S, and 16S genes. Maximum parsimony, maximum likelihood, and Bayesian inference analyses were used for phylogenetic reconstruction and divergence time estimation analyses. Intra- and interspecific genetic distances were calculated among species using the COI dataset. The results showed that at least three of the five currently accepted species are well defined: P. fulvinota, P. pseudolyrcea, and P. paraensis, however, a significant level of intraspecific variation was observed in P. fulvinota. These findings will assist future revisions of the description, classification, and distribution of species of Paralucilia, as well as in the elaboration of taxonomic keys. Additionally, we show that it is possible to clarify the evolutionary history of this Neotropical genus using supplementary evidence such as morphology and molecular data.

The Mesembrinellidae (Diptera: Oestroidea) comprise a small group of strictly Neotropical calyptrate flies, with 36 described species. The group has often been treated as a subfamily of Calliphoridae, but there is growing evidence that it corresponds to a distinct Oestroidea lineage. Internal relationships have so far been addressed based only on morphology, with results lacking resolution and support. This is the first molecular phylogeny for the group, which is based on the analyses of 80 terminal taxa (22 mesembrinellid and 28 outgroup species) and 5 molecular markers (ITS2, 28S, COI, COII and 16S). Maximum-parsimony, maximum-likelihood and Bayesian inference methods were used, the latter two with partitioning strategies considering codon position and secondary structure information. Results corroborate the Mesembrinellidae as a monophyletic lineage inside Oestroidea. Three clades were consistently recovered: (1) (Laneella + Mesembrinella patriciae); (2) (Mesembrinella (excluding M. patriciae) + Eumesembrinella); and (3) (Huascaromusca + Giovanella). Re-examination of the female reproductive tract of M. patriciae revealed a Laneela-type spermatheca, which corroborates the position of the species recovered in the molecular phylogenetic analyses. Mesembrinella and Huascaromusca are in all cases paraphyletic with regards to Eumesembrinella and Giovanella, respectively. These latter two genera should, thus, be seen as subjective junior synonyms.

BackgroundThe genotype-phenotype interactions among traits governing feeding preference are of fundamental importance to behavioral genetics and evolutionary biology. The genetic basis of behavioral traits has been explored in different taxa using different approaches. However, the complex nature of the genetic mechanisms undergirding behavior is poorly understood. Here, we present an evolutionary study of candidate genes related to parasitism in Calliphoridae (Diptera: Calyptratae). Closely related species in this family exhibit distinct larval feeding habits, most notably necro-saprophagy and obligate parasitism.MethodsTo understand the genetic and molecular bases underlying these habits, expression levels of eight candidate genes for feeding behavior—Cyp6g2, foraging, glutamate dehydrogenase, Jonah65aiv, Malvolio, PGRP-SC2,RPS6-p70-protein kinase,andsmooth—were measured in four species using qPCR. Moreover we used expression values and sequence information to reconstruct the relationship among species and thedN/dSrate to infer possible sites under selection.ResultsFor most candidate genes, no statistically significant differences were observed, indicating a high degree of conservation in expression. However,Malvoliowas differentially expressed between habits. Evolutionary analyses based on transcript levels and nucleotide sequences ofMalvoliocoding region suggest that transcript levels were correlated to feeding habit preferences among species, although deviations under a strictly neutral model were also observed in statistical tests.DiscussionMalvoliowas the only gene demonstrating a possible connection to feeding habit. Differences in gene expression may be involved in (or be a result of) the genetic regulation of Calliphoridae feeding habit. Our results are the first steps towards understanding the genetic basis and evolution of feeding behavior in Calliphoridae using a functional approach.

True flies are insects of the order Diptera and encompass one of the most diverse groups of animals on Earth. Within dipterans, Schizophora represents a recent radiation of insects that was used as a model to develop a pipeline for generating complete mitogenomes using various sequencing platforms and strategies. 91 mitogenomes from 32 different species were sequenced and assembled with high fidelity, using amplicon, whole genome shotgun or single molecule sequencing approaches. Based on the novel mitogenomes, we estimate the origin of Schizophora within the Cretaceous-Paleogene (K-Pg) boundary, about 68.3 Ma. Detailed analyses of the blowfly family (Calliphoridae) place its origin at 22 Ma, concomitant with the radiation of grazing mammals. The emergence of ectoparasitism within calliphorids was dated 6.95 Ma for the screwworm fly and 2.3 Ma for the Australian sheep blowfly. Varying population histories were observed for the blowfly Chrysomya megacephala and the housefly Musca domestica samples in our dataset. Whereas blowflies (n = 50) appear to have undergone selective sweeps and/or severe bottlenecks in the New World, houseflies (n = 14) display variation among populations from different zoogeographical zones and low levels of gene flow. The reported high-throughput mitogenomics approach for insects enables new insights into schizophoran diversity and population history of flies.