Background: Blowflies are ubiquitous insects, often shiny and metallic, and the larvae of many species provide important ecosystem services (e.g., recycling carrion) and are used in forensics and debridement therapy. Yet, the taxon has repeatedly been recovered to be para- or polyphyletic, and the lack of a well-corroborated phylogeny has prevented a robust classification.Results: We here resolve the relationships between the different blowfly subclades by including all recognized subfamilies in a phylogenomic analysis using 2221 single-copy nuclear protein-coding genes of Diptera. Maximum likelihood (ML), maximum parsimony (MP), and coalescent-based phylogeny reconstructions all support the same relationships for the full data set. Based on this backbone phylogeny, blowflies are redefined as the most inclusive monophylum within the superfamily Oestroidea not containing Mesembrinellidae, Mystacinobiidae, Oestridae, Polleniidae, Sarcophagidae, Tachinidae, and Ulurumyiidae. The constituent subfamilies are re-classified as Ameniinae (including the Helicoboscinae,syn. nov.), Bengaliinae, Calliphorinae (including Aphyssurinae,syn. nov., Melanomyinae,syn. nov., and Toxotarsinae,syn. nov.), Chrysomyinae, Luciliinae, Phumosiinae, Rhiniinaestat. rev., and Rhinophorinaestat. rev. Metallic coloration in the adult is shown to be widespread but does not emerge as the most likely ground plan feature.Conclusions: Our study provides the first phylogeny of oestroid calyptrates including all blowfly subfamilies. This allows settling a long-lasting controversy in Diptera by redefining blowflies as a well-supported monophylum, and blowfly classification is adjusted accordingly. The archetypical blowfly trait of carrion-feeding maggots most likely evolved twice, and the metallic color may not belong to the blowfly ground plan.

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.

Stomach bot flies (Calyptratae: Oestridae, Gasterophilinae) are obligate endoparasitoids of Proboscidea (i.e. elephants), Rhinocerotidae (i.e. rhinos) and Equidae (i.e. horses and zebras, etc.), with their larvae developing in the digestive tract of hosts with very strong host specificity. They represent an extremely unusual diversity among dipteran, or even insect parasites in general, and therefore provide significant insights into the evolution of parasitism. The phylogeny of stomach bot flies was reconstructed based on extensive mitochondrial genomic data for Cobboldia , Gyrostigma and six of the eight known species of Gasterophilus . The phylogenetic tree, i.e. { Cobboldia , [ Gyrostigma , ( Gasterophilus pecorum , ( Gasterophilus intestinalis , ( Gasterophilus haemorrhoidalis , Gasterophilus inermis )), ( Gasterophilus nasalis , Gasterophilus nigricornis ))]}, provides a strong evolutionary reference to infer several biological patterns for the first time for this group: (i) host shifts of stomach bot flies from elephants to rhinoceroses and then from rhinoceroses to equids; (ii) dispersal with their hosts from the Afrotropical region into the Palaearctic and Oriental regions; (iii) oviposition site, originally on the host head, and egg production positively correlated with distance from the mouth; (iv) attachment of third‐instar larva originally in the stomach, with duodenal and large intestinal positions secondarily derived; and (v) guanine and cytosine enrichment of the mitogenome as an adaptation to larval life in the warm environment of the host digestive tract, combined with the need for a high evolutionary rate to cope with the fast evolution of their mammalian hosts.