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Toxotarsinae is a subfamily of Calliphoridae (Diptera: Calyptratae: Oestroidea) with 11 species currently distributed in three genera: Neta Shannon, 1926, Sarconesia Bigot, 1857, and Toxotarsus Macquart, 1851. All known species are endemic to South America, mostly restricted to areas with cooler temperatures—like high elevation areas of the Andean Cordillera—and lowlands and coastal regions in the subtropical and temperate parts of the continent. The classification of the subfamily has been somewhat controversial, with eleven, mostly monotypic nominal genera established to accommodate its species, mostly due to distinct interpretations of certain controversial characters by different authors. To provide a sound basis for a generic classification, we performed the first study aimed to understand the transformation series of those controversial characters and character states under a phylogenetic framework. Our maximum parsimony analyses, under both equal and implied weighting of characters and including all currently known species in the subfamily, retrieved two main clades of Toxotarsinae: the roraima group, composed of Sarconesia roraima (Townsend, 1935), Neta chilensis (Walker, 1836), and S. magellanica (Le Guillou, 1842), and the splendida group, composed of the remaining species of the subfamily. The position of S. magellanica varied, but the species was mostly recovered within the roraima group except in the analyses under implied weighting with k values between 1 and 6. Our results, which include an explicit interpretation of the transformation series associated with some characters and character states commonly used in Toxotarsinae systematics, suggest that both Sarconesia and Neta, along with other genera proposed in the past for species of the subfamily such as Sarconesiomima Lopes & Albuquerque, 1955, Chlorobrachycoma Townsend, 1918, Sarconesiopsis Townsend, 1918, and Roraimomusca Townsend, 1935, previously suggested to be synonyms of Sarconesia, do not properly reflect the evolution of lineages in the subfamily and should not be considered as valid nominal genera. Toxotarsus was the only genus consistently recovered as monophyletic and, based on the phylogenetic relationships recovered herein, we suggest including all known Toxotarsinae within this genus, with the roraima and splendida clades treated as species-groups. Finally, we discuss the characters commonly used to delimit the taxa subordinated to Toxotarsinae, such as the length of the first flagellomere, the position of the rays of the arista relative to the pedicel, the extension of the plumosity of the arista, and the number of postsutural acrostichal setae.

Measuring species richness of tropical forests is a major challenge. Such measurement is a key information in many senses, from an evolutionary perspective to conservation of threatened, fragile habitats. Data has gradually shown that the canopy of tropical forest is a hugely complex component of the forest, but a precise assessment of the diversity of megadiverse groups in the canopy is still wanting. We collected large samples of insects were along a period of two weeks using 6-meter Gressitt-style Malaise traps set at five heights on a metal tower in a tropical forest north of Manaus—one trap at the ground level, one trap above the canopy (32 m) and three traps at intermediate levels (8, 16 and 24 m). The samples contained 37,778 specimens belonging to 18 order of insects. Fifty-seven families of flies (Diptera) were found, 39 of which were identified to 368 genera and 856 species. The species of these 39 families of flies fit into eight patterns of vertical distribution of abundance and species richness of the fauna, with patterns of one or two peaks of species at different levels. A total of 527 (61.6%) of the 856 fly species recognized in the samples were not collected at the ground level. The canopy-associated species of Diptera showed a high species richness and a relatively low abundance indicating that they represent vulnerable components of tropical diversity. The biology of the flies families and genera we collected in the canopy suggest that the evolution of flies went through a unique process: independent clades of Diptera explored in different ways the resources originated along the very evolution of the angiosperm forest canopy along the early Cenozoic. Unlike other primarily phytophagous groups of insects, flies radiated into a large array of biologies associated with the canopies: parasitoids, hematophagous, phytosaprophagous, kleptoparasites, sap feeders, gall-makers etc. The results only stress the need of additional strategies to protect this diversity.

The Calyptratae, one of the most species-rich fly clades, only originated and diversified after the Cretaceous-Palaeogene extinction event and yet exhibit high species diversity and a diverse array of life history strategies including predation, phytophagy, saprophagy, haematophagy and parasitism. We present the first phylogenomic analysis of calyptrate relationships. The analysis is based on 40 species representing all calyptrate families and on nucleotide and amino acid data for 1456 single-copy protein-coding genes obtained from shotgun sequencing of transcriptomes. Topologies are overall well resolved, robust and largely congruent across trees obtained with different approaches (maximum parsimony, maximum likelihood, coalescent-based species tree, four-cluster likelihood mapping). Many nodes have 100% bootstrap and jackknife support, but the true support varies by more than one order of magnitude [Bremer support from 3 to 3427; random addition concatenation analysis (RADICAL) gene concatenation size from 10 to 1456]. Analyses of a Dayhoff-6 recoded amino acid dataset also support the robustness of many clades. The backbone topology Hippoboscoidea+(Fanniidae+(Muscidae+((Anthomyiidae-Scathophagidae)+Oestroidea))) is strongly supported and most families are monophyletic (exceptions: Anthomyiidae and Calliphoridae). The monotypic Ulurumyiidae is either alone or together with Mesembrinellidae as the sister group to the rest of Oestroidea. The Sarcophagidae are sister to Mystacinobiidae+Oestridae. Polleniinae emerge as sister group to Tachinidae and the monophyly of the clade Calliphorinae+Luciliinae is well supported, but the phylogenomic data cannot confidently place the remaining blowfly subfamilies (Helicoboscinae, Ameniinae, Chrysomyinae). Compared to hypotheses from the Sanger sequencing era, many clades within the muscoid grade are congruent but now have much higher support. Within much of Oestroidea, Sanger era and phylogenomic data struggle equally with regard to finding well-supported hypotheses.

In this paper we improve the morphological description of the tree cricket Neoxabea brevipes Rehn, 1913 and a new occurrence record for the species at the São Francisco de Paula National Forest Conservation Unit, State of Rio Grande do Sul, southern Brazil. Morphological characterization is provided together with illustrations emphasizing the most significant diagnostic features, such as the metanotal gland, abdominal specializations and phallic sclerites. The calling song was field recorded, comprising a continuous trill with dominant frequency of 3.6 kHz (19.7°C). The karyotype was described for the first time to Neoxabea Kirby, 1906, including diploid number of 2n=26, with two sexual chromosomes.

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.

The Mesembrinellidae (Diptera: Oestroidea) are a group of exclusively Neotropical flies, with 38 recognized species distributed in nine genera. A total of 11 of these species, present in seven genera, were known to occur in Colombia. Huascaromusca obscura Wolff, sp. nov. and H. cordillera Wolff & Ramos-Pastrana, sp. nov. are herein described and their male terminalia are illustrated. Based on a prior molecular study of the family, Giovanella carvalhoi Wolff et al., 2013 is transferred to Huascaromusca Townsend. A key to all known species of Huascaromusca is provided.

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.