Revision of the Brazilian species of Ptilodexia Brauer & Bergenstamm, 1889 (Diptera: Tachinidae: Dexiinae), with the description of a new species.

The Brazilian species of the New World genus Ptilodexia Brauer & Bergenstamm, 1889 are revised. Before this study, only one species of Ptilodexia was recorded from Brazil, viz. P. lateralis (Walker, 1836). Herein we record, for the first time, two new records of known Ptilodexia species in Brazil, viz. P. striata (Wulp, 1891) and P. rubricornis (Wulp, 1891). In addition, a new species is described, Ptilodexia matogrossensis sp. nov. from Mato Grosso and Mato Grosso do Sul states in Brazil; hence four species of this genus are currently recorded from Brazil. The species Neomyostoma ptilodexioides Townsend, 1935, from Brazil, placed in the monotypic genus Neomyostoma Townsend, 1935, is proposed as junior synonym of P. lateralis. Illustrations and detailed descriptions are presented for P. lateralis, P. matogrossensis sp. nov., P. striata and P. rubricornis and the male terminalia is described for P. lateralis and P. striata. The female terminalia and the first larval instar are described for the first time for the genus, based on the descriptions of P. lateralis and P. striata. A key to the identification of all recognized Brazilian species of Ptilodexia is presented. Finally, an updated distributional record is given for all studied species.

Carrion insects living within the bones of large mammals: insect conservation and forensic entomology implications.

Succession patterns of carrion insects on large mammal's carrion has been widely studied, notably to estimate the post-mortem interval in forensic investigations as accurately as possible. However, little attention has been paid to the carrion insects living inside these bones once a carcass is skeletonized. One very recent study documented flies emerging from pig carcasses, and only scarce authors reported the presence of other carrion insects taking advantage of the bone marrow. We, thus, aimed to (1) estimate the frequency of inner-bone space colonization by carrion insects, with particular attention to bone-skipper flies; (2) identify the insects living inside the carrion bones; and (3) determine whether or not carrion insects found within the bones can successfully exit the bones and complete their development. We extensively sampled 185 large mammals' bones collected from twelve vulture feeding stations and four isolated carcasses in southwest France and northern Spain. Sampled bones were opened, and the insects found inside were identified. For two bones, foramen, i.e., the holes providing a natural entrance and exit to the bone's inner cavity, was monitored with a camera to assess the insect's putative exit. We describe the entomofauna, i.e., the set of insect species, living within the bones, and illustrate insects' ability to exit the bones for their subsequent development and maturity. These results are discussed in the framework of carrion insect conservation and forensic entomology perspectives.

Eocene amber provides the first fossil record and bridges distributional gap in the rare genus Robsonomyia (Diptera: Keroplatidae).

Until now, the genus Robsonomyia was represented by two extant species: R. reducta Matile & Vockeroth, 1980 from North America and R. sciaraeformis (Okada, 1939) from Asia. This paper presents the first fossil members of the genus Robsonomyia, which is also the first record from Europe. Two new fossil species from Baltic amber are described: R. baltica Pelczynska, Krzeminski & Blagoderov, sp. nov. and R. henningseni Pelczynska, Krzeminski & Blagoderov, sp. nov.. The presence of fossil Robsonomyia spp. on the European continent suggests Holarctic distribution of the genus in the past. We also discuss possible pathways of its intercontinental dispersion.

Two new species of Dasybasis Macquart from the Peruvian Andes of Cusco (Diptera: Tabanidae: Diachlorini).

Two new horse fly species, Dasybasis mancoi sp. nov. and D. huaynai sp. nov. (Diptera: Tabanidae: Diachlorini), are described from the Peruvian Andes near Cusco on the basis of morphological features, including those of the female genitalia. Digital photographs of both species and of the terminalia of both females and of one male are provided. The two new species are related to D. bulbula Coscarn & Philip, as evidenced by their similar morphological characteristics.

On the immature stages of some Merodontini hoverflies (Diptera: Syrphidae) from Europe and Africa.

The genera Eumerus and Merodon (Syrphidae: Merodontini) form together the most speciose grouping of hoverflies in the Palaearctic Region. However, little is known about the morphology and biology of their larvae. The few larvae of Eumerus and Merodon that have been uncovered are phytophagous in underground organs of plants (some Eumerus and all Merodon) or saprophagous in a variety of plants' parts (the reminder of Eumerus). In this study, the second larval stage (L2) of Eumerus lyneborgi Ricarte & Hauser, 2020 and both the larva (L2) and puparium of Merodon constans (Rossi, 1794) are described for the first time. Larvae of E. lyneborgi were found in a decaying stem of Cyphostemma juttae (Dinter & Gilg) Desc., 1960 (Vitaceae) in Namibia (Africa), while larvae of M. constans were collected in bulbs of Leucojum vernum Linnaeus, 1753, (Amaryllidaceae) in France (Europe). Morphology of the immature forms was studied by observation and imaging with stereomicroscopy and scanning electron microscopy. The head skeleton of E. lyneborgi larvae was found to be of the filter feeding type, i.e., in accordance with a saprophagous trophic regime, while that of M. constans was typically phytophagous. Variability in certain characters of the M. constans early stages is described and discussed in relation to the adult form and molecular information published in literature. An updated identification key to all known third larval stages/puparia of Merodon is provided.

Morphological studies of fly puparia (Diptera: Calliphoridae) in the soil from a Joseon Dynasty grave in Korea.

Archaeoentomological investigations were conducted on soil contents from a grave belonging to the Joseon Dynasty as part of the Urban Environment Maintenance Project (UEMP) in Cheongjin 12-16 dong (districts), Jongno-gu, Seoul, Korea, from December 01, 2008 to February 19, 2011. A total of 28 insect puparia with hard shells of the common green bottle fly Lucilia sericata were identified in the soil. Evidence suggested that the corpse was placed outside for some days instead of being buried immediately after death. This is the first report of fly puparia in soil samples from a tomb of the Joseon Dynasty during 16-17 AD in Korea. Our findings may help determine the timeframe of burial and offer archaeological insights into the funerary customs of the period.

A new species of Mycomya Rondani, 1856 (Diptera: Mycetophilidae) common to southeast Australia, Tasmania, and New Zealand.

Mycomya quadrimaculata sp. nov. is described from specimens collected in southeast Australia, Tasmania, and New Zealand. A key and type photographs of known Australian and New Zealand Mycomya species are provided. The relative abundance, observed distribution, and morphological affinities of the new species suggests that it is adventive and a recent introduction to New Zealand. Wing characters indicate that the new species is most closely aligned with a subgroup of the Australian Mycomya fauna.

A new species of Lucilia Robineau-Desvoidy (Diptera, Calliphoridae) from the Amazon region and its potential distribution areas.

Lucilia amazonia, sp. nov. from the Amazon region of Colombia and Brazil is described based on morphology. This species appears to be limited to the Amazon region and is quite distinctive and unlike other Lucilia in the region. A key to the species is provided following the Lucilia key in Whitworth (2014). Based on the distribution observed, its likely distribution is predicted.

Size rather than complexity of sexual ornaments prolongs male metamorphosis and explains sexual size dimorphism in sepsid flies.

Male sexual ornaments often evolve rapidly and are thought to be costly, thus contributing to sexual size dimorphism. However, little is known about their developmental costs, and even less about costs associated with structural complexity. Here, we quantified the size and complexity of three morphologically elaborate sexually dimorphic male ornaments that starkly differ across sepsid fly species (Diptera: Sepsidae): (i) male forelegs range from being unmodified, like in most females, to being adorned with spines and large cuticular protrusions; (ii) the fourth abdominal sternites are either unmodified or are converted into complex de novo appendages; and (iii) male genital claspers range from small and simple to large and complex (e.g. bifurcated). We tracked the development of 18 sepsid species from egg to adult to determine larval feeding and pupal metamorphosis times of both sexes. We then statistically explored whether pupal and adult body size, ornament size and/or ornament complexity are correlated with sex-specific development times. Larval growth and foraging periods of male and female larvae did not differ, but the time spent in the pupal stage was ca 5% longer for sepsid males despite emerging 9% smaller than females on average. Surprisingly, we found no evidence that sexual trait complexity prolongs pupal development beyond some effects of trait size. Evolving more complex traits thus does not incur developmental costs at least in this system.

Larval anatomy of the digestive and excretory systems of the pharyngeal bot fly, Pharyngomyia picta (Diptera: Oestridae).

Oestrid flies (Diptera: Oestridae) are obligate parasites of mammals during their larval stage and show anatomical adaptations for the infestation of host tissues. Unlike the species that parasitize domestic mammals, those oestrid species that infest wild mammal hosts remain poorly known. With the use of x-ray micro-computed tomography, we describe for the first time the anatomy of the digestive and excretory systems of the second and third larval instars of Pharyngomyia picta (Meigen), a parasite of cervids that, like other species within the subfamily Oestrinae, causes nasopharyngeal myiasis. Both larval instars of P. picta show a pair of remarkably large salivary glands arranged in a characteristic 'glandular band', a convoluted and thickly uniform midgut and a greatly enlarged distal region of the anterior pair of Malpighian tubules. These anatomical features also have been described in other species within the subfamily Oestrinae, whereas they differ from the observations in other oestrid subfamilies. We discuss the potential functional significance of the anatomy of the digestive and excretory systems of Oestrinae larvae as specific adaptations to parasitize the nasopharyngeal cavities of mammal hosts.

Morphological and molecular data reveal fifteen new sympatric Zygomyia Winnertz, 1864 (Diptera: Mycetophilidae) species from Ecuador.

Fifteen new Zygomyia Winnertz species are described from material collected from one location in Ecuador during a very short time period: Z. acrotricha sp. nov., Z. comata sp. nov., Z. diversiseta sp. nov., Z. flamma sp. nov., Z. geniculata sp. nov., Z. longistyla sp. nov., Z. napoensis sp. nov., Z. paniculata sp. nov., Z. pectinella sp. nov., Z. spinosa sp. nov., Z. talpaeforma sp. nov., Z. trilobata sp. nov., Z. triseta sp. nov., Z. uniordata sp. nov. and Z. ximoi sp. nov. Zygomyia bicolor Edwards, 1934 is redescribed, including detailed illustration of the male terminalia. The number of Neotropical Zygomyia species is increased to 30. An updated key to the Neotropical Zygomyia is provided. Unexpectedly high level of interspecific genetic variation between Neotropical Zygomyia spp. is reported.

Species of Paraclius Loew (Diptera: Dolichopodidae) with bi-lamellate antennal stylus: new combination, new synonym, and two new species.

The species of Paraclius Loew with bi-lamellate arista-like stylus are revised and P. americanus (Schiner, 1868) comb. nov. is transferred from Tachytrechus Haliday and regarded as the senior synonym of P. diplacocerus Capellari, 2013 syn. nov. In addition, two new species with similar antennal morphology are described and illustrated from Brazil: P. bilamellatus sp. nov. (States of Espírito Santo and Goiás) and P. vulcanoae sp. nov. (State of São Paulo). A distribution map and a key to the males of known species of Paraclius with bi-lamellate arista-stylus are provided.

Morphological and molecular characterization of puparia of Piophilidae species of forensic relevance.

Piophilidae are a small family of Diptera with a worldwide distribution and which are historically associated with human activities. In addition to their economic importance, piophilid larvae can also be of medical and legal relevance. Within a medicolegal context, piophilids are frequently associated with cadavers in advanced stages of decomposition, thus being potentially useful forensic indicators and they have been reported also from archaeo-funerary contexts. An accurate species identification is therefore an essential prerequisite to ensure the reliable analysis of insect material in medical, forensic and archaeological investigations. Identification of the adult piophilid flies is possible because of the availability of identification keys, in contrast immature insects, especially puparia, have been poorly investigated and described. In this paper, puparia of 11 species of forensic interest (Piophila casei, Piophila megastigmata, Parapiophila atrifrons, Parapiophila flavipes, Parapiophila vulgaris, Protopiophila litigata, Liopiophila varipes, Prochyliza nigrimana, Prochyliza xanthosoma and Stearibia nigriceps in subtribe Piophilina and Centrophlebomyia furcata in subtribe Thyreophorina) are described and a molecular analysis, based on the COI sequencing, is presented to show the potential of the molecular approach in their identification.

A new genus of dance fly (Diptera: Empidoidea: Hybotidae) from Cretaceous Spanish ambers and introduction to the fossiliferous amber outcrop of La Hoya (Castellón Province, Spain).

Hybotidae fly species, also known as dance flies, in Cretaceous ambers have been described from Lebanon, France, Myanmar, Russia, and Canada. Here we describe Grimaldipeza coelica gen. et sp. n., and recognize another two un-named species, in Spanish amber from the middle Albian El Soplao and lower Cenomanian La Hoya outcrops. The fore tibial gland is present in the new genus, which is characteristic of the family Hybotidae. We compare Grimaldipeza coelica gen. et sp. n. with the holotypes of Trichinites cretaceus Hennig, 1970 and Ecommocydromia difficilis Schlüter, 1978, and clarify some morphological details present in the latter two species. Further taxonomic placement beyond family of the here described new genus was not possible and remains incertae sedis within Hybotidae until extant subfamilies are better defined. We provide new paleoecological data of the hybotids, together with paleogeographical and life paleoenvironmental notes. A table with the known Cretaceous Hybotidae is provided. Furthermore, the La Hoya amber-bearing outcrop is described in detail, filling the information gap for this deposit.

Larvae and adult flies of Rhinoestrus purpureus and R. usbekistanicus: morphology and pupation (Diptera: Oestridae).

Rhinoestrus species larvae are considered obligatory parasites of the nasal cavities of equine. This type of myiasis is characterised by sneezing, coughing, olfactory nerve damage and encephalomyelitis. Also, it has a zoonotic importance as the larvae might cause ophthalmomyiasis and conjunctivitis in human. While few studies describing R. purpureus adult fly antennal sensillae are available, the R. usbekistanicus antennal sensillae have never been described. Also, scanty data are available on the adult flies of Rhinoestrus species morphology. For this reason, the current study aimed at identiying and comparing different Rhinoestrus species (larvae, adult flies and time of pupation). Using light and scanning electron microscopy, we have evidenced differences between R. purpureus, R. usbekistanicus larvae in spination pattern, shape of spines, peritremes shape and ultrastrucures. The study also showed that for both species the pupa required 15­21 days at room temperature to develop into adult flies, identified the adult male flies and female external genitalia have been identified. As well, the gross features of R. purpureus and R. usbekistanicus adult flies which included the disposition of the parafrontalia and parafascialia tubercles, mesonotal weals, wings and abdominal pellonisityhave been characterised and the sensillae compared.

Sphenempis, a new Empis subgenus (Diptera, Empididae) from the Eastern Palaearctic Region.

Empis (Sphenempis Saigusa) subgen. nov. (Diptera, Empididae) and its included species, Empis (Sphenempis) bicoloriventris Saigusa sp. nov. from the Ryukyus, Japan and Empis (Sphenempis) oshimai Saigusa sp. nov. from Kyushu, Japan are described. Empis (Lissempis) amurensis Shamshev, 1998 from Amur, Russia is newly included in the subgenus Sphenempis.

A new species of Procanace Hendel from the Philippines (Diptera: Canacidae: Canacinae).

A new species of surf flies (Canacidae: Canacinae) is described and illustrated based on four specimens (1 , 3 ) collected by Dr. Amnon Freidberg in the Philippines. Some peculiarities of the postabdominal structures, especially those of the male terminalia, suggest a fairly close affinity with two other congeneric species, namely Procanace townesi Wirth, 1951, and P. cogani Mathis, 1988, having a disjointed distribution, Marshall Islands and Seychelles, respectively. It is further suggested that these affinities should arguably be regarded as synapomorphic character states in the groundplan of the genus.

Wing measurements are a possible tool for the identification of European forensically important Sarcophagidae.

The flesh flies are a group of insects well known for their forensic importance. Reliable identification of these flies relies on the use of either molecular markers or the morphology of the male genital apparatus. Identification of female flesh flies is more time consuming and less reliable than their male counterparts. This is particularly problematic for forensic entomology practitioners, because female flesh flies are more abundant than males in carrion arthropod assemblages. As such, it is critical that alternative methods for flesh fly identification are established that are equally effective for both sexes. One promising technique is the use of wing measurements, which have been shown to be reliable for the identification of some groups of necrophagous Diptera from several geographical regions. We applied this method to the European Sarcophagidae for the first time, using a dataset of 881 specimens representing 29 species and 5 genera. Species identifications were based on 15 landmarks located at wing vein junctions. We also combined our results with data from previous studies of Calliphoridae and Muscidae which utilised the same methodology, enabling the testing of family level identification using wing morphometrics. Species identifications using wing measurements had varied success. While some species were successfully identified without error, others, particularly from the genus Sarcophaga, were often misclassified. Notably, in several species wing measurements successfully identified both males and females. The results presented here suggest that wing measurements are a promising complementary method to other methods for the identification of necrophagous Sarcophagidae especially in material unsorted at the family level. It can also be used to double check identification performed by a taxonomist using traditional methods.

Sperm competition in yellow dung flies: No consistent effect of sperm size.

The male competition for fertilization that results from female multiple mating promotes the evolution of increased sperm numbers and can impact sperm morphology, with theory predicting that longer sperm can at times be advantageous during sperm competition. If so, males with longer sperm should sire more offspring than competitors with shorter sperm. Few studies have directly tested this prediction, and findings are inconsistent. Here we assessed whether longer sperm provide a competitive advantage in the yellow dung fly (Scathophaga stercoraria; Diptera: Scathophagidae). Initially, we let brothers with different temperature-mediated mean sperm lengths compete - thus minimizing confounding effects of genetic background - and found no clear advantage of longer sperm. We then used flies from lines subjected to bidirectional selection on phenoloxidase activity that had shown correlated evolutionary responses in sperm and female spermathecal duct lengths. This experiment also yielded no main effect of sperm size on siring success. Instead, there was a trend for a shorter-sperm advantage, but only when competing in females with longer spermathecal ducts. Our data corroborated many previously reported findings (last-male precedence, effects of copula duration and body size), suggesting our failure to find sperm size effects is not inherently due to our experimental protocols. We conclude that longer sperm are not competitively superior in yellow dung flies under most circumstances, and that, consistent with previous work, in this species competitive fertilization success is primarily determined by the relative numbers of sperm competing.

The Acquisition and Retention of Lumpy Skin Disease Virus by Blood-Feeding Insects Is Influenced by the Source of Virus, the Insect Body Part, and the Time since Feeding.

Lumpy skin disease virus (LSDV) is a poxvirus that causes severe systemic disease in cattle and is spread by mechanical arthropod-borne transmission. This study quantified the acquisition and retention of LSDV by four species of Diptera (Stomoxys calcitrans, Aedes aegypti, Culex quinquefasciatus, and Culicoides nubeculosus) from cutaneous lesions, normal skin, and blood from a clinically affected animal. The acquisition and retention of LSDV by Ae. aegypti from an artificial membrane feeding system was also examined. Mathematical models of the data were generated to identify the parameters which influence insect acquisition and retention of LSDV. For all four insect species, the probability of acquiring LSDV was substantially greater when feeding on a lesion compared with feeding on normal skin or blood from a clinically affected animal. After feeding on a skin lesion LSDV was retained on the proboscis for a similar length of time (around 9 days) for all four species and for a shorter time in the rest of the body, ranging from 2.2 to 6.4 days. Acquisition and retention of LSDV by Ae. aegypti after feeding on an artificial membrane feeding system that contained a high titer of LSDV was comparable to feeding on a skin lesion on a clinically affected animal, supporting the use of this laboratory model as a replacement for some animal studies. This work reveals that the cutaneous lesions of LSD provide the high-titer source required for acquisition of the virus by insects, thereby enabling the mechanical vector-borne transmission. IMPORTANCE Lumpy skin disease virus (LSDV) is a high consequence pathogen of cattle that is rapidly expanding its geographical boundaries into new regions such as Europe and Asia. This expansion is promoted by the mechanical transmission of the virus via hematogenous arthropods. This study quantifies the acquisition and retention of LSDV by four species of blood-feeding insects and reveals that the cutaneous lesions of LSD provide the high titer virus source necessary for virus acquisition by the insects. An artificial membrane feeding system containing a high titer of LSDV was shown to be comparable to a skin lesion on a clinically affected animal when used as a virus source. This promotes the use of these laboratory-based systems as replacements for some animal studies. Overall, this work advances our understanding of the mechanical vector-borne transmission of LSDV and provides evidence to support the design of more effective disease control programmes.