Phylogenetic relationships within the oestroid subclades Rhinophorinae (Calliphoridae) and Polleniidae were reconstructed for the first time, applying a Sanger sequencing approach using the two protein-coding nuclear markers CAD (carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase; 1794 bp) and MCS (molybdenum cofactor sulfurase; 2078 bp). Three genera of Polleniidae and nineteen genera of Rhinophorinae were analyzed together with a selection of taxa representing the major lineages of Oestroidea (non-rhinophorine Calliphoridae, Oestridae, Sarcophagidae, Tachinidae). The selected markers provide good resolution and moderate to strong support of the distal branches, but weak support for several deeper nodes. Polleniidae (cluster flies) emerge as monophyletic and their sister-group relationship to Tachinidae is confirmed. Morinia Robineau-Desvoidy as currently circumscribed emerges as paraphyletic with regard to Melanodexia Williston, and Pollenia Robineau-Desvoidy is the sister taxon of the Morinia-Melanodexia clade. We propose a classification with two subfamilies, Moriniinae Townsend (including Morinia, Melanodexia, and Alvamaja Rognes), and Polleniinae Brauer & Bergenstamm (including Pollenia, Dexopollenia Townsend, and Xanthotryxus Aldrich). Anthracomyza Malloch and Nesodexia Villeneuve are considered as Oestroidea incertae sedis pending further study. Rhinophorinae (woodlouse flies) emerge as monophyletic and sister to a clade composed of (Ameniinae + (Ameniinae + Phumosiinae)), and a tribal classification is proposed with the subfamily divided into Rhinophorini Robineau-Desvoidy, 1863 and Phytonini Robineau-Desvoidy, 1863 (the Stevenia-group and the Phyto-group of authors, respectively). Oxytachina Brauer & Bergenstamm, 1891, stat. rev. is resurrected to contain nine Afrotropical rhinophorine species currently assigned to genus Rhinomorinia Brauer & Bergenstamm, 1891: Oxytachina approximata (Crosskey, 1977) comb. nov., O. atra (Bischof, 1904) comb. nov., O. bisetosa (Crosskey, 1977) comb. nov., O. capensis (Brauer & Bergenstamm, 1893) comb. nov., O. scutellata (Crosskey, 1977) comb. nov., O. setitibia (Crosskey, 1977) comb. nov., O. verticalis (Crosskey, 1977) comb. nov., O. vittata Brauer & Bergenstamm, 1891, and O. xanthocephala (Bezzi, 1908) comb. nov.

Farming insects has recently emerged as a new source of protein and lipid production. To date, research has mostly focused on food applications of insects. Focusing on nonfood potential of oil and proteins of insects, high-throughput studies of insect lipids and proteins are needed. We performed proteomics and lipidomics investigation on black soldier fly (Hermitia illucens) and blow fly (Lucilia sericata) larvae to investigate new potential and applications. We used mass spectrometry for proteomics and lipidomics analysis of control and treated larvae. Treatment was performed by incubation with a biological decomposer. We provide the list of all fatty acids with their concentration in control and treated larvae. This result showed high levels of lauric acid in black soldier fly, which could even increase after biological decomposition. Proteomics analysis showed the presence of proteins like collagen of cosmetic interest, and proteins with antimicrobial properties such as phenoloxidases and enzymatic activities, such as amylase and trypsin. Insects harbor high potential for nonfood usage as additives, antimicrobial effects, and even pharmaceuticals and cosmetics. These data open avenues for future research in pharmacological and cosmetic approaches to find new molecules of interests.

The control of onchocerciasis in the African region is currently based mainly on the mass drug administration of ivermectin. Whilst this has been found to limit morbidity, it does not stop transmission. In the absence of a macrofilaricide, there is a need for an integrated approach for disease management, which includes vector control. Vector control using chemical insecticides is expensive to apply, and therefore the use of other measures such as biological control agents is needed. Immature stages of Simulium squamosum, reared in the laboratory from egg masses collected from the field at Boti Falls and Huhunya (River Pawnpawn) in Ghana, were observed to be attacked and fed upon by larvae of the chironomid Cardiocladius oliffi Freeman, 1956 (Diptera: Chironomidae).

The complete mitogenome of the horse stomach bot fly Gasterophilus pecorum (Fabricius) and a near-complete mitogenome of Wohlfahrt's wound myiasis fly Wohlfahrtia magnifica (Schiner) were sequenced. The mitogenomes contain the typical 37 mitogenes found in metazoans, organized in the same order and orientation as in other cyclorrhaphan Diptera. Phylogenetic analyses of mitogenomes from 38 calyptrate taxa with and without two non-calyptrate outgroups were performed using Bayesian Inference and Maximum Likelihood. Three sub-analyses were performed on the concatenated data: (1) not partitioned; (2) partitioned by gene; (3) 3rd codon positions of protein-coding genes omitted. We estimated the contribution of each of the mitochondrial genes for phylogenetic analysis, as well as the effect of some popular methodologies on calyptrate phylogeny reconstruction. In the favoured trees, the Oestroidea are nested within the muscoid grade. Relationships at the family level within Oestroidea are (remaining Calliphoridae (Sarcophagidae (Oestridae, Pollenia + Tachinidae))). Our mito-phylogenetic reconstruction of the Calyptratae presents the most extensive taxon coverage so far, and the risk of long-branch attraction is reduced by an appropriate selection of outgroups. We find that in the Calyptratae the ND2, ND5, ND1, COIII, and COI genes are more phylogenetically informative compared with other mitochondrial protein-coding genes. Our study provides evidence that data partitioning and the inclusion of conserved tRNA genes have little influence on calyptrate phylogeny reconstruction, and that the 3rd codon positions of protein-coding genes are not saturated and therefore should be included.

The world Rhinophoridae are catalogued, recognising 33 genera and 177 species. Nomenclatural information is provided for all genus-group and species-group names, including lists of synonyms and name-bearing type data. Species distributions are recorded by country. A key to the world genera is presented. Four new genera are erected to accommodate five new species, which do not fit within any of the current generic concepts in Rhinophoridae, according to the results of a morphology-based phylogenetic analysis: Marshallicona Cerretti & Pape with type species Marshallicona quitu Cerretti & Pape, gen. et sp. nov. (Ecuador); Maurhinophora Cerretti & Pape with type species Maurhinophora indoceanica Cerretti & Pape, gen. et sp. nov. (Mauritius); Neotarsina Cerretti & Pape with type species Neotarsina caraibica Cerretti & Pape, gen. et sp. nov. (Trinidad and Tobago) and Neotarsina andina Cerretti & Pape, sp. nov. (Peru); Kinabalumyia Cerretti & Pape with type species Kinabalumyia pinax Cerretti & Pape, gen. et sp. nov. (Malaysia, Sabah). The genus Aporeomyia Pape & Shima (type species Aporeomyia antennalis Pape & Shima), originally assigned to Tachinidae, is here reassigned to Rhinophoridae based on a reassessment of the homologies of the male terminalia. The following five species-group names, which were previously treated as junior synonyms or nomina dubia, are recognised as valid species names: Acompomintho caucasica (Villeneuve, 1908), stat. rev. [from nomen dubium to valid species]; Acompomintho sinensis (Villeneuve, 1936), stat. rev. [from nomen dubium to valid species]; Stevenia bertei (Rondani, 1865), stat. rev. [from nomen dubium to valid species]; Stevenia sardoa Villeneuve, 1920, stat. rev. [from junior synonym of Rhinophora deceptoria Loew, 1847 to valid species]; Stevenia subalbida (Villeneuve, 1911), stat. rev. [from junior synonym of Rhinophora deceptoria Loew, 1847 to valid species]. Reversal of precedence is invoked for the following case of subjective synonymy to promote stability in nomenclature: Rhinophora lepida (Meigen, 1824), nomen protectum, and Musca parcus Harris, 1780: 144, nomen oblitum. New generic and specific synonymies are proposed for the following two names: Mimodexia Rohdendorf, 1935, junior synonym of Tromodesia Rondani, 1856, syn. nov. and Ptilocheta tacchetti Rondani, 1865, junior synonym of Stevenia obscuripennis (Loew, 1847), syn. nov. The following new combinations are proposed: Acompomintho sinensis (Villeneuve, 1936), comb. nov. [transferred from Tricogena Robineau-Desvoidy, 1830]; Tromodesia guzari (Rohdendorf, 1935), comb. nov. [transferred from Mimodexia Rohdendorf, 1935]; Tromodesia intermedia (Rohdendorf, 1935), comb. nov. [transferred from Mimodexia Rohdendorf, 1935]; Tromodesia lindneriana (Rohdendorf, 1961), comb. nov. [transferred from Mimodexia Rohdendorf, 1935]; Tromodesia magnifica (Rohdendorf, 1935), comb. nov. [transferred from Mimodexia Rohdendorf, 1935]; Tromodesia obscurior (Rohdendorf, 1935), comb. nov. [transferred from Mimodexia Rohdendorf, 1935]; Tromodesia pallidissima (Rohdendorf, 1935), comb. nov. [transferred from Mimodexia Rohdendorf, 1935]; Tromodesia setiventris (Rohdendorf, 1935), comb. nov. [transferred from Mimodexia Rohdendorf, 1935] and Tromodesia shachrudi (Rohdendorf, 1935), comb. nov. [transferred from Mimodexia Rohdendorf, 1935].

The presence of nosocomial myiasis reflects a lack of adequate medical attention, due to the physical facilities and/or the health care personnel. Patients requiring special attention are more susceptible, such as those with a loss of consciousness, assisted mechanical ventilation, tracheal tubes, or nasogastric probes. Nosocomial myiasis is a rare event that has a greater occurrence in the hospitals of poor and developing countries. The two cases herein described represent the first report of nosocomial myiasis in Mexico. The causal agents were found to be Lucilia sericata and Sarcophaga spp. The taxonomical identification of the larvae of the second and third instar was based on the morphology of the cephaloskeleton, anterior spiracles, and peritreme plaques.

In light of the African swine fever outbreaks in Estonian pig farms during the past few years, the question of the vector potential of Diptera in the pig farm environment has risen. However, the arthropod fauna of the pig farm environment is currently not well established. Hence, the aim of this study was to clarify the species diversity in pig farms. In total, 22 Diptera species or species groups were found in Estonian pig farms. There were altogether 186,701 individual arthropods collected, from which 96.6% (180,444) belonged to the order of true flies (Insecta: Diptera). The remaining 3.4% were from other insect orders, arachnids, or just damaged and unidentifiable specimens. The activity density and diversity of dipterans differed significantly between 12 sampled farms but not throughout the sampling period. The present study is amongst the few to provide a large-scale overview of pig-farm-associated Diptera in the temperate climate zone.

Marine dwelling in Diptera has been relatively unexplored and the frequency of transitions to the marine environment and the evolutionary history remain poorly understood. By reviewing records from the World Register of Marine Species and using ancestral state reconstruction methods, we build on the fly tree of life phylogeny and ecological descriptions of marine life history. Our ancestral state reconstruction analyses suggest marine dwelling is lacking as an ancestral trait for the most recent common ancestor to Diptera. While many transitions in Empidoidea, Sciomyzoidea, Tipulomorpha, and Culicomorpha seem to have been gradual, other transitions in Tephritoidea and Tabanomorpha were found likely to have been stochastic occurrences. From the collection of 532 marine species, we reveal several independent transitions to the marine environment throughout the fly tree of life. Considering the results from our analysis, we outline potential adaptations for marine flies and discuss the barriers of colonizing the marine environment and the implications to the mechanisms for salt tolerance.

Many species groups, including mammals and many insects, determine sex using heteromorphic sex chromosomes. Diptera flies, which include the model Drosophila melanogaster, generally have XY sex chromosomes and a conserved karyotype consisting of six chromosomal arms (five large rods and a small dot), but superficially similar karyotypes may conceal the true extent of sex chromosome variation. Here, we use whole-genome analysis in 37 fly species belonging to 22 different families of Diptera and uncover tremendous hidden diversity in sex chromosome karyotypes among flies. We identify over a dozen different sex chromosome configurations, and the small dot chromosome is repeatedly used as the sex chromosome, which presumably reflects the ancestral karyotype of higher Diptera. However, we identify species with undifferentiated sex chromosomes, others in which a different chromosome replaced the dot as a sex chromosome or in which up to three chromosomal elements became incorporated into the sex chromosomes, and others yet with female heterogamety (ZW sex chromosomes). Transcriptome analysis shows that dosage compensation has evolved multiple times in flies, consistently through up-regulation of the single X in males. However, X chromosomes generally show a deficiency of genes with male-biased expression, possibly reflecting sex-specific selective pressures. These species thus provide a rich resource to study sex chromosome biology in a comparative manner and show that similar selective forces have shaped the unique evolution of sex chromosomes in diverse fly taxa.

The 16 world species of Sphecomyia Latreille are revised, including seven previously undescribed species (S.cryptica Moran, sp. n., S.hoguei Moran, sp. n., S.interrupta Moran, sp. n., S.oraria Moran, sp. n., S.pseudosphecomima Moran, sp. n., S.sexfasciata Moran, sp. n., and S.weismani Moran, sp. n.). Descriptions, redescriptions, male genitalia photographs, distribution maps, and an illustrated key for all Sphecomyia are presented. DNA barcode data are provided for all 16 species with a cytochrome oxidase subunit I gene tree presented and discussed. Sphecomyia stat. rev. is redefined to represent the monophyletic lineage of species within subtribe Criorhinina possessing a bare, medial vitta extending ventrally from the oral margin in both sexes, a bare gena, a bare katepimeron, a scutellum with at least anterior margin densely pruinose, an anterior ventral half of vein C before crossvein h without setae, and a narrow intersection of vein R1 with vein C. Three species groups of Sphecomyia are identified: the S.vittata group which possess pruinose scutellar vittae, the S.pattonii group which lack pruinose scutellar vittae, and S.metallica (Bigot), a hairy bee mimic with a completely pruinose scutum. Criorhinatsherepanovi Violovitsh is resurrected and transferred, along with Criorhinaaino Stackelberg, to the genus Sphecomyia: S.tsherepanovi (Violovitsh), comb. n. and S.aino (Stackelberg), comb. n. Criorhinametallica (Bigot) is designated as the senior synonym of C.lupina (Williston), not junior as improperly treated, and transferred to Sphecomyia: S.metallica (Bigot), comb. n. The species Sphecomyiafusca Weisman, S.nasica Osburn, and S.occidentalis Osburn are transferred to Criorhina Meigen: C.fusca (Weisman), comb. n., C.nasica (Osburn), comb. n., and C.occidentalis (Osburn), comb. n.

Exechia is a diverse genus of small fungus gnats, widespread in the Holarctic Region, while the fauna is largely unknown elsewhere, such as in the Afrotropical and Oriental Region. Members of Exechia can be arranged into several species groups, based on homologies in the male and female terminalia. The Exechiaparva group is delimited, based on male terminalia possessing a pair of gonocoxal lobes on the apicoventral gonocoxal margin. Eight previously-described species can be placed in this group, of which six are from the Holarctic Region, while one is recorded each from the Oriental and the Afrotropical Regions.

Although Aedes fluviatilis is an anthropophilic mosquito found abundantly in urban environments, its biology, epidemiological potential and genetic characteristics are poorly understood. Climate change and urbanization processes that result in environmental modifications benefit certain anthropophilic mosquito species such as Ae. fluviatilis, greatly increasing their abundance in urban areas. To gain a better understanding of whether urbanization processes modulate the genetic structure of this species in the city of São Paulo, we used eight microsatellite loci to genetically characterize Ae. fluviatilis populations collected in nine urban parks in the city of São Paulo. Our results show that there is high gene flow among the populations of this species, heterozygosity deficiency and low genetic structure and that the species may have undergone a recent population expansion. There are two main hypotheses to explain these findings: (i) Ae. fluviatilis populations have undergone a population expansion as a result of urbanization; and (ii) as urbanization of the city of São Paulo occurred recently and was quite intense, the structuring of these populations cannot be observed yet, apart from in the populations of Ibirapuera and Piqueri parks, where the first signs of structuring have appeared. We believe that the expansion found in Ae. fluviatilis populations is probably correlated with the unplanned urbanization of the city of São Paulo, which transformed green areas into urbanized areas, as well as the increasing population density in the city.

Aedes aegypti is one of the species most favored by changes in the environment caused by urbanization. Its abundance increases rapidly in the face of such changes, increasing the risk of disease transmission. Previous studies have shown that mosquito species that have adapted to anthropogenic environmental changes benefit from urbanization and undergo population expansion. In light of this, we used microsatellite markers to explore how urbanization processes may be modulating Ae. aegypti populations collected from three areas with different levels of urbanization in the city of São Paulo, Brazil. Specimens were collected at eleven sites in three areas with different degrees of urbanization in the city of São Paulo: conserved, intermediate and urbanized. Ten microsatellite loci were used to characterize the populations from these areas genetically. Our findings suggest that as urbanized areas grow and the human population density in these areas increases, Ae. aegypti populations undergo a major population expansion, which can probably be attributed to the species' adaptability to anthropogenic environmental changes. Our findings reveal a robust association between, on the one hand, urbanization processes and densification of the human population and, on the other, Ae. aegypti population structure patterns and population expansion. This indicates that this species benefits from anthropogenic effects, which are intensified by migration of the human population from rural to urban areas, increasing the risk of epidemics and disease transmission to an ever-increasing number of people.

Phorid flies are amongst the most biologically diverse and species-rich groups of insects. Ways of life range from parasitism, herbivory, fungivory, to scavenging. Although the lifestyles of most species are unknown, many are parasitoids, especially of social insects. Some species of ant-parasitoids are attracted to injured hosts for feeding purposes to develop eggs, as well as for oviposition, requiring each female to find two injured hosts.

The horn fly, Haematobia irritans (L.), is an obligate hematophagous ectoparasite of cattle, and one of the most important pests of cattle causing unrealized gains or losses in meat and milk production. The present study describes the difficulties that arise when research programs have attempted to maintain this pest, both on-host and off-host, in a laboratory environment. Suggestions aimed at assisting future researchers in successfully colonizing horn flies in the laboratory are provided.

Aedes aegypti mosquitoes are capable of vectoring a wide range of diseases including dengue, yellow fever, and Zika viruses, with approximately half of the worlds' population at risk from such diseases. Development of combined predator-parasite treatments for the control of larvae consistently demonstrates increased efficacy over single-agent treatments, however, the mechanism behind the interaction remains unknown. Treatments using the natural predator Toxorhynchites brevipalpis and the entomopathogenic fungus Metarhizium brunneum were applied in the laboratory against Ae. aegypti larvae as both individual and combined treatments to determine the levels of interaction between control strategies. Parallel experiments involved the removal of larvae from test arenas at set intervals during the course of the trial to record whole body caspase and phenoloxidase activities. This was measured via luminometric assay to measure larval stress factors underlying the interactions. Combined Metarhizium and Toxorhynchites treatments were seen to drastically reduce lethal times as compared to individual treatments. This was accompanied by increased phenoloxidase and caspase activities in combination treatments after 18 h (p < 0.001). The sharp increases in caspase and phenoloxidase activities suggest that combined treatments act to increase stress factor responses in the larvae that result in rapid mortality above that of either control agent individually. This work concludes that the underlying mechanism for increased lethality in combined parasite-predator treatments may be related to additive stress factors induced within the target host larvae.

Culex quinquefasciatus mosquitoes can be found in almost every major city of Brazil and are vectors of filariasis and several arboviruses. Microsatellite markers have been widely used to uncover the genetic structure of various groups of insect populations. The aim of this study was to glimpse the genetic structure of Cx. quinquefasciatus in Brazil.

The long-legged fly Asyndetus clavipes belongs to the subfamily Diaphorinae of Dolichopodidae. The mitogenome of A. clavipes was sequenced, the first representative complete mitogenome from this subfamily. This mitogenome is 15,385 bp in size, includes 13 protein-coding genes, 22 transfer RNAs, and two ribosomal RNAs. All genes have the same location and coding strand as in other published species of Dolichopodidae. Nucleotide composition is biased toward A and T, which together made up 78.6% of the entire genome. Bayesian inference strongly supported the monophyly of Empidoidea, Empididae and Dolichopodidae, with the phylogenetic relationships within Empidoidea: ((Dolichopodinae + Neurigoninae) + Diaphorinae) + ((Trichopezinae + (Empidinae + Oreogetoninae)) + Ocydromiinae).

The long-legged fly Xanthochlorus tibetensis belongs to the subfamily Xanthochlorinae of Dolichopodidae. The mitogenome of X. tibetensis was sequenced, it is the first representative complete mitogenome from this subfamily. This mitogenome is 15,580 bp in size, includes 13 protein-coding genes, 22 transfer RNAs, and two ribosomal RNAs. All genes have the same location and coding strand as in other published species of Dolichopodidae. Nucleotide composition is biased toward A and T, which together made up 77.6% of the entire genome. Bayesian inference strongly supported the monophyly of Empidoidea, Empididae and Dolichopodidae, with the phylogenetic relationships within Empidoidea: ((Dolichopodinae + Xanthochlorinae) + Neurigoninae) + ((Trichopezinae + (Empidinae + Oreogetoninae)) + Ocydromiinae).

The complete mitochondrial genome of Parastratiosphecomyia szechuanensis Lindner, 1954 was sequenced and analyzed in this study. The mitochondrial genome is 16,414 bp in length, including the 37 typical insect mitochondrial genes and a large control region. All PCGs end with complete termination codon TAA or TAG. Most PCGs initiated by standard start codon ATN, except for cox1 which starts with TCG. The phylogenetic analysis based on the nucleotide sequence data of 13 PCGs recovered the monophyly of Stratiomyidae and the sister relationship between Xylomyidae and Stratiomyidae.