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To address the limited number of mitochondrial genomes (mitogenomes) in the subfamily Steganinae (Diptera: Drosophilidae), we assembled 12 complete mitogenomes for six representative species in the genus Amiota and six representative species in the genus Phortica. We performed a series of comparative and phylogenetic analyses for these 12 Steganinae mitogenomes, paying special attention to the commonalities and differences in the D-loop sequences. Primarily determined by the lengths of the D-loop regions, the sizes of the Amiota and Phortica mitogenomes ranged from 16,143–16,803 bp and 15,933–16,290 bp, respectively. Our results indicated that the sizes of genes and intergenic nucleotides (IGNs), codon usage and amino acid usage, compositional skewness levels, evolutionary rates of protein-coding genes (PCGs), and D-loop sequence variability all showed unambiguous genus-specific characteristics and provided novel insights into the evolutionary implications between and within Amiota and Phortica. Most of the consensus motifs were found downstream of the D-loop regions, and some of them showed distinct genus-specific patterns. In addition, the D-loop sequences were phylogenetically informative as the data sets of PCGs and/or rRNAs, especially within the genus Phortica.

With the increasing number of cryptic species discovered in major branches of the subfamily Steganinae, accurate species identification is becoming challenging. Possible evolutionary stasis often gives rise to the formation of morphological conservatism and cryptic species, as is the case for the members of the Stegana (subgen. Orthostegana). In this study, the species boundaries of all Asian Orthostegana were accessed based on two mitochondrial genes [cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 2 (ND2)] using different species delimitation methods and evolutionary models. By integrating morphological and molecular evidence, we clearly validated the evolutionary independence of all five known species and identified nine new species from Southwest China: Stegana (Orthostegana) aini Peng & Chen sp. nov., S. (O.) brevivittata Peng & Chen sp. nov., S. (O.) cuodi Peng & Chen sp. nov., S. (O.) fuscofemorata Peng & Chen sp. nov., S. (O.) latipalpula Peng & Chen sp. nov., S. (O.) macrostephana Peng & Chen sp. nov., S. (O.) mohnihei Peng & Chen sp. nov., S. (O.) obscurala Peng & Chen sp. nov., and S. (O.) pinguitia Peng & Chen sp. nov. Moreover, we discussed the relationship between the Asian Orthostegana lineage and the subgen. Oxyphortica, and the necessity of taxonomic re-evaluation for the Neotropical species with molecular data. The high morphological conservatism within Orthostegana is largely the outcome of in situ diversification for adapting to the ecological environment.

A new species belonging to the previously monotypic steganine genus Mayagueza Wheeler, 1960 (Diptera: Drosophilidae) is described. The new species, Mayagueza lopesi sp. nov., was collected in the Brazilian Cerrado. This is the first record of this genus for South America, and the description of the new species, based on male and female adult specimens, includes photographs and detailed illustrations of male and female terminalia.

BackgroundThe Drosophilidae family is traditionally divided into two subfamilies: Drosophilinae and Steganinae. This division is based on morphological characters, and the two subfamilies have been treated as monophyletic in most of the literature, but some molecular phylogenies have suggested Steganinae to be paraphyletic. To test the paraphyletic-Steganinae hypothesis, here, we used genomic sequences of eight Drosophilidae (three Steganinae and five Drosophilinae) and two Ephydridae (outgroup) species and inferred the phylogeny for the group based on a dataset of 1,028 orthologous genes present in all species (> 1,000,000 bp). This dataset includes three genera that broke the monophyly of the subfamilies in previous works. To investigate possible biases introduced by small sample sizes and automatic gene annotation, we used the same methods to infer species trees from a set of 10 manually annotated genes that are commonly used in phylogenetics.ResultsMost of the 1,028 gene trees depicted Steganinae as paraphyletic with distinct topologies, but the most common topology depicted it as monophyletic (43.7% of the gene trees). Despite the high levels of gene tree heterogeneity observed, species tree inference in ASTRAL, in PhyloNet, and with the concatenation approach strongly supported the monophyly of both subfamilies for the 1,028-gene dataset. However, when using the concatenation approach to infer a species tree from the smaller set of 10 genes, we recovered Steganinae as a paraphyletic group. The pattern of gene tree heterogeneity was asymmetrical and thus could not be explained solely by incomplete lineage sorting (ILS).ConclusionsSteganinae was clearly a monophyletic group in the dataset that we analyzed. In addition to ILS, gene tree discordance was possibly the result of introgression, suggesting complex branching processes during the early evolution of Drosophilidae with short speciation intervals and gene flow. Our study highlights the importance of genomic data in elucidating contentious phylogenetic relationships and suggests that phylogenetic inference for drosophilids based on small molecular datasets should be performed cautiously. Finally, we suggest an approach for the correction and cleaning of BUSCO-derived genomic datasets that will be useful to other researchers planning to use this tool for phylogenomic studies.

Two conspicuous Steganinae species, Stegana (Steganina) magnifica Hendel, 1913 from Amazonian Peru and Stegana (Ceratostylus) fumipennis (Enderlein, 1922) from southern Brazil, are redescribed based on holotypes, and their identities are clarified. Both species are exclusive to the Neotropical Region and the first, with a body length of about 5.5 mm, is the largest species of Stegana described so far in this region, while the latter displays a peculiar antenna bearing an unusual, forward-projected, comma-shaped flagellomere 1. The photomicrographs of the habitus and terminalia of each specimen are also provided.

The taxonomy of the subgenus Ashima of the genus Phortica is revised. A cladistic analysis of 66 morphological characters is conducted, covering 35 species (28 known and 7 new species: Phortica efragmentata sp. nov., P. andreagigoni sp. nov., P. watabei sp. nov., P. halimunensis sp. nov., P. akutsui sp. nov., P. kerinciensis sp. nov., and P. takehiroi sp. nov.) which correspond to 71.4% of 49 total spp. of Ashima. The resulting cladogram shows that the studied species are separated into several clades/subclades/cluster each highly supported with specific synapomorphies. Those clades/subclades/cluster are newly defined as the following species groups, subgroups or complex: the afoliolata, foliiseta, nigrifoliiseta and angulata species groups; the foliiseta, tanabei, nigrifoliiseta and glabra species subgroups; and the foliiseta species complex. The subgenus Ashima is peculiar in having the asymmetric male genitalia as the ground plan and showing the antisymmetry (i.e. intraspecific mirror-image variation) in some species but the directional asymmetry (i.e. side-fixed asymmetry) in others. The evolution of genital asymmetry in this subgenus is estimated by mapping the states (symmetry, directional asymmetry and antisymmetry) of bilateral structures of male genitalia on the cladogram. This ancestral state reconstruction estimates that the directional asymmetry of male genitalia has evolved at the ancestor of this subgenus and then changed to the antisymmetric state independently in two lineages, the angulata + nigrifoliiseta species groups and the foliiseta species complex. In this study, a standardized terminology recently proposed for the male terminalia of Drosophila melanogaster is extendedly adopted to describe the morphology of male terminalia in the subgenus Ashima, one group of the subfamily Steganinae.

Six new species of the genus Phortica are described from Yunnan, Southwest China: P. (Alloparadisa) kaibangya n. sp., P. (Ashima) semiannulata n. sp., P. (Ashima) dianmianensis n. sp., P. (Ashima) ...

Rhinoleucophenga Hendel comprises an unusual Drosophilidae (Diptera) genus with predaceous larvae, currently compounded by 29 nominal species with New World distribution. In the present study, Rhinoleucophenga brasiliensis (Costa Lima) and R. fluminensis (Costa Lima) are redescribed. These two species are commonly misidentified in Drosophilidae species inventories, mainly by the few morphological character details presented in the original taxonomic description. Thus, by the morphological review performed here, lectotype and paralectotypes designed to R. brasiliensis and R. fluminensis, as well as new morphological characters, drawings and photos (for the first time) are presented in order to avoid further taxonomic mistakes with those referred sibling species of Rhinoleucophenga.

Background The fruit fly Phortica variegata (Drosophilidae: Steganinae) feeds on the ocular secretions of animals and humans, and has been described as an intermediate host of the eye worm Thelazia callipaeda (Spirurida: Thelaziidae) in Italy. Despite the increased detection of T. callipaeda in many European countries, information about its vector role in natural conditions is still limited. In the Iberian Peninsula, thelaziosis caused by T. callipaeda has been reported in dogs, cats, red foxes, wild rabbits and humans. Methods In the last seven years, we have detected increased numbers of cases of canine thelaziosis at three locations in mainland Spain: Site 1, La Vera region (Cáceres Province, central-western Spain; 51 cases); Site 2, El Escorial municipality (Madrid Community, central Spain; 23 cases); and Site 3, Miraflores de la Sierra municipality (Madrid Community, central Spain; 41 cases). Site 1 is considered endemic for T. callipaeda while the other two sites have been recently recognised as risk zones for T. callipaeda infection. Results From June 2016 to September 2017, 2162 flies were collected and morphologically identified as Phortica spp. (Site 1, n = 395; Site 2, n = 1544; and Site 3, n = 223). Upon dissection, third-stage T. callipaeda larvae were found in two out of 155 flies examined from Site 1, and both these larvae tested molecularly positive for the eye worm. Of the 395 flies collected from Site 1, 371 were molecularly processed for arthropod species identification and T. callipaeda detection. All 371 flies were identified as P. variegata and 28 (7.5%; 95% CI: 4.8–10%) tested positive for T. callipaeda DNA haplotype 1. Conclusions Our findings indicate that T. callipaeda circulates among dogs and P. variegata in Spain, where zoonotic cases have been also reported. The co-existence of canine thelaziosis and Phortica spp. in geographical areas previously considered free of the eye worm indicates a risk of infection for both animals and humans living in this region.

A total of 50 (43 known and seven new) species in the subgenus Phortica (sensu stricto) were surveyed and (re)described from China: P. bicornuta (Chen & Toda, 1997); P. bipartita (Toda & Peng, 1992); P. biprotrusa (Chen & Toda, 1998); P. cardua (Okada, 1977); P. chi (Toda & Sidorenko, 1996); P. conifera (Okada, 1977); P. eparmata (Okada, 1977); P. eugamma (Toda & Peng, 1990); P. excrescentiosa (Toda & Peng, 1990); P. fangae (Máca, 1993); P. flexuosa (Zhang & Gan, 1986); P. foliata (Chen & Toda, 1997); P. gamma (Toda & Peng, 1990); P. gigas (Okada, 1977); P. glabtabula Chen & Gao, 2005; P. hainanensis (Chen & Toda, 1998); P. hongae (Máca, 1993); P. huazhii Cheng & Chen, 2008; P. iota (Toda & Sidorenko, 1996); P. jadete Zhu, Cao & Chen, 2018; P. kappa (Máca, 1977); P. lambda (Toda & Peng, 1990); P. latifoliacea Chen & Watabe, 2008; P. magna (Okada, 1960); P. okadai (Máca, 1977); P. omega (Okada, 1977); P. orientalis (Hendel, 1914); P. pangi Chen & Wen, 2005; P. paramagna (Okada, 1971); P. perforcipata (Máca & Lin, 1993); P. pi (Toda & Peng, 1990); P. protrusa (Zhang & Shi, 1997); P. pseudopi (Toda & Peng, 1990); P. pseudotau (Toda & Peng, 1990); P. psi (Zhang & Gan, 1986); P. rhagolobos Chen & Gao, 2008; P. saeta (Zhang & Gan, 1986); P. setitabula Chen & Gao, 2005; P. subradiata (Okada, 1977); P. tau (Toda & Peng, 1990); P. uncinata Chen & Gao, 2005; P. unipetala Chen & Wen, 2005; P. allomega Gong & Chen, sp. nov.; P. archikappa Gong & Chen, sp. nov.; P. dianzangensis Gong & Chen, sp. nov.; P. imbacilia Gong & Chen, sp. nov.; P. liukuni Gong & Chen, sp. nov.; P. tibeta Gong & Chen, sp. nov.; and P. xianfui Gong & Chen, sp. nov. In addition, seven new synonyms were recognized: P. acongruens (Zhang & Shi, 1997), syn. nov.; P. antillaria (Chen & Toda, 1997), syn. nov.; P. kukuanensis Máca, 2003, syn. nov.; P. linae (Máca & Chen, 1993), syn. nov.; P. shillongensis (Singh & Gupta, 1979), syn. nov.; P. takadai (Okada, 1977), syn. nov.; and P. watanabei (Máca & Lin, 1993), syn. nov. A key to all Asian species (except for the eparmata species complex) of this subgenus was provided. All currently available DNA barcode (partial mitochondrial cytochrome c oxidase subunit I (COI) gene) sequences of this subgenus (217 sequences of 54 species) are employed in a molecular analysis using different species delimitation methods. The results indicate that approximately 68.5% (37 of 54 spp.) of Phortica (s. str.) species could be clearly distinguished from closely related morphospecies or cryptic species.

For slightly more than a century only the type specimen has been known for Pyrgometopa penicillata, a monotypic genus of Drosophilidae whose sole species has an ocellar triangle distended into a bizarre tubercle bearing a tuft of large, spine-like bristles in both sexes. Here we report additional specimens of both sexes of the species, recently rediscovered from Brazil, Peru and French Guyana, allowing us to state that Pyrgometopa is a junior synonym for Stegana, a large cosmopolitan genus. A detailed redescription of Stegana penicillata (Kertész) new combination is provided, including new morphological information, like the male and female terminalia, the egg and a short discussion is provided about its hallmark feature.

Rhinoleucophenga Hendel is an endemic genus of the New World with most species recorded in Brazil. Rhinoleucophenga obesa (Loew) seemed to be the most widespread species, being recorded in the United States of America, Mexico and Brazil. In the Neotropical region, identifications of R. obesa were commonly based on the description of non-type specimens determined by Costa Lima (1935) and Malogolowkin (1946), although their identities were doubtful. However, the recent redescription of R. obesa from the type-series from Texas, USA, confirmed a long period of misidentifications and a new species, R. cantareira Vilela & Bächli, was proposed based on Brazilian specimens. Thus, review of morphological and molecular traits of specimens previously identified as R. obesa in Neotropical biomes are necessary to check the identity and distribution of a group of sibling species: R. obesa, R. cantareira, R. gigantea (Thomson), R. pallida Hendel and R. pampeana Poppe et al. In the present paper, specimens previously determined as R. obesa, sampled in different Brazilian localities, were compared by morphological and molecular traits. Plates of female terminalia of R. cantareira and R. gigantea are presented for the first time. The spermathecal capsules (as well as the male epandrium) revealed useful characteristics to differentiate those species; these morphological differences were corroborated by a sequence fragment of COI. The specimens on which were based the descriptions of Rhinoleucophenga obesa sensu Costa Lima (1935) and sensu Malogolowkin (1946) were reviewed and confirmed as R. cantareira. Other Brazilian specimens from different localities, previously misidentified as R. obesa, were determined as R. gigantea or R. cantareira, with new records for both species. Therefore, the present study corresponds to the advance of identity definition and distribution of sibling species of Rhinoleucophenga commonly sampled in Neotropical inventory studies.

Rhinoleucophenga pallida Hendel, 1917 (type species of the genus) is redescribed based on its female holotype and a male from a nearby locality, and Rhinoleucophenga obesa (Loew, 1872) on its two syntypes, which are designated as the male lectotype and a female paralectotype. Both are valid species. A proposal is made to establish the genus Pseudophortica Sturtevant, 1918 (type species R. obesa), a junior synonym of Rhinoleucophenga, to subgenus rank and include all species of Rhinoleucophenga described or redescribed from males except R. pallida, which is unique in having a remarkable pedunculate surstylus, among other differences. The North American R. obesa is compared to its closest sibling, the South American species Rhinoleucophenga gigantea (Thomson, 1869). The occurrence of R. obesa in Brazil is also questioned, as suggested long ago by Marshall R. Wheeler. The specimens from Brazil previously identified as such most probably belong to the new species described in the present paper as Rhinoleucophenga (Pseudophortica) cantareira sp. nov. (type locality: Parque Estadual da Cantareira, City of São Paulo, State of São Paulo, Brazil). Numerous photomicrographs of their habitus and male terminalia taken with a Smartphone's rear camera and digitally stacked to create images with greater depth of focus are provided. Keywords: Habitus, Illustrations, Lectotype designation, Steganinae, Terminalia

The Neotropical region harbors an astonishing diversity of species, but still encompasses the least studied biogeographic region of the world. These properties apply for different taxonomic groups, and can be exemplified by drosophilids. In fact, high levels of cryptic diversity have recently been discovered for Neotropical species of the Zygothrica genus group, but relationships among these species, or them and other Drosophilidae species still remains to be addressed. Therefore, the aim of this study was to evaluate the phylogenetic relationships between fungus-associated Neotropical species of the genera Hirtodrosophila, Mycodrosophila and Zygothrica, which together with Paramycodrosophila and Paraliodrosophila compose the Zygothrica genus group. For this, fragments of the mitochondrial cytochrome oxidase subunits I (COI) and II (COII) genes, and the nuclear alpha methyldopa (Amd) and dopa decarboxylase (Ddc) genes were newly characterized for 43 Neotropical specimens of fungus-associated drosophilids, and analyzed in the context of 51 additional Drosophilinae sequences plus one Steganinae outgroup. Based on the resulting phylogeny, the evolution of breeding sites usage was also evaluated through ancestral character reconstructions. Our results revealed the Zygothrica genus group as a monophyletic lineage of Drosophila that branches after the subgenera Sophophora and Drosophila. Within this lineage, Mycodrosophila species seem to encompass the early offshoot, followed by a grade of Hirtodrosophila species, with derived branches mostly occupied by representatives of Zygothrica. This genus, in particular, was subdivided into five major clades, two of which include species of Hirtodrosophila, whose generic status needs to be reevatuated. According to our results, the use of fungi as breeding sites encompasses a symplesiomorphy for the Zygothrica genus group, since one of the recovered clades is currently specialized in using flowers as breeding sites whereas a sole species presents a reversal to the use of fruits of a plant of Gentianales. So, in general, this study supports the paraphyly of Drosophila in relation to fungus-associated Neotropical species of Drosophilidae, providing the first molecular insights into the phylogenetic patterns related to the evolution of this diverse group of species and some of its characteristic traits.

A new species of Rhinoleucophenga Hendel from the Brazilian Amazon, R. hesperides sp. nov., was described based on morphology of male and female specimens. The larvae are predators of the eggs and adult females of ensign scales Praelongorthezia praelonga (Douglas) (Hemiptera: Sternorrhyncha: Ortheziidae). Female P. praelonga also serve as a source of honeydew for adult R. hesperides sp. nov. In addition, the puparium of this species was described.

Knowledge of the effects that Thelazia callipaeda (Spirurida, Thelaziidae) infection has on the survival of its vector Phortica variegata (Drosophilidae, Steganinae) is scarce. The present study aimed to: (a) assess the rate of infection between experimentally infected (EI) and not experimentally infected (NEI) flies and (b) determine how T. callipaeda infection may affect the survival of P. variegata. In addition, fat composition was evaluated in flies that died during overwintering. Molecular analysis showed that T. callipaeda prevalence in flies that died before experimental infection, plus those from the NEI group, is 0.75% (i.e. 11 out of 1462 individuals). The EI group showed a significantly higher positivity to T. callipaeda (i.e. 51 out of 682 individuals; 7.48%) compared with the NEI group (i.e. 9 out of 750 individuals; 1.2%). Thelazia callipaeda DNA was detected until 147 days after experimental infection. This demonstrates that larvae of this eyeworm may survive in the fly for a significant period of the winter. Fat composition analysis showed that flies produced more unsaturated than saturated fatty acids during diapause, probably because unsaturated fatty acids remain in a liquid state at lower temperatures, providing anti-freeze properties to survive winter.

Rhinoleucophenga myrmecophaga sp. nov., a Drosophilidae species associated with extrafloral nectaries of Qualea grandiflora Mart. (Vochysiaceae) and whose larvae are mostly predaceous on ants, is described from specimens collected in the Cerrado biome of Itirapina, state of Sao Paulo, Brazil. Illustrations of imagos, male terminalia, and immatures are also included. This is the first evidence of dipteran larvae that are closely associated with extrafloral nectaries and that feed on visiting ants.

The phylogenetic relationships among the East Asian species of the apodemata and sinuata species groups of the genus Amiota were investigated based on DNA sequences of the mitochondrial NADH dehydrogenase subunit 2 (ND2) gene. A total of 23 samples of 12 species were employed as in-group taxa, and one sample for each of four other Amiota species were used as out-groups. The results suggested with strong confidence the monophyly of both the apodemata and the sinuata groups, whereas the monophyly of the ‘apodemata group + sinuata group’ cluster was less supported. Based on its geographical distribution, the origin of the sinuata group is supposed to be southern China. Four new species were described from Guangxi and Yunnan, China: Amiota reikae Xu & Chen sp. nov., Amiota guiensis Xu & Chen sp. nov., Amiota hesongensis Xu & Chen sp. nov., and Amiota polytreta Xu & Chen sp. nov. © 2013 The Linnean Society of London

Molecular sequences now overwhelm morphology in phylogenetic inference. Nonetheless, most molecular studies are conducted on a limited number of taxa, as DNA rarely can be analysed from old museum types or fossils. During the last 20 years, more than 150 molecular studies have challenged the current phylogenetic classification of the family Drosophilidae Rondani based on morphological characters. Most studies concerned a single genus, Drosophila Fallen, and included only few representative species from 17 out of the 78 genera of the family. Therefore, these molecular studies were unable to provide an alternative classification scheme. A supermatrix analysis of seven nuclear and one mitochondrial genes (8248 bp) for 33 genera was conducted using outgroups from one calyptrate and four ephydroid families. The Bayesian phylogeny was consistent with previous molecular studies including whole genome sequences and divided the Drosophilidae into four monophyletic clades. Morphological characters, mostly male genitalia, then were compared thoroughly between the four clades and homologous character states were identified. These states were then checked for 70 genera and a revised phylogenetic, family-group classification for the Drosophilidae is proposed. Two genera –Cladochaeta Coquillett and Diathoneura Duda – of the tribe Cladochaetini Grimaldi are transferred to the family Ephydridae. The Drosophilidae is divided into two subfamilies: Steganinae Hendel (30 genera) and Drosophilinae Rondani (43 genera). A further two genera, Apacrochaeta Duda and Sphyrnoceps de Meijere, are incertae sedis, and Palmophila Grimaldi, is synonymized with Drosophilasyn.n. The Drosophilinae is subdivided into two tribes: the re-elevated Colocasiomyini Okada (nine genera) and Drosophilini Okada. The paraphyly of the genus Drosophila was not resolved to avoid affecting the binomina of important laboratory model species; however, its subgeneric classification was revised in light of molecular and morphological data. Three subgenera, namely Chusqueophila Brncic, Phloridosa Sturtevant and Psilodorha Okada, were synonymized with the subgenus Drosophila (Drosophila) Fallen syns.n. Among the 45 species groups and 5 species complexes of Drosophila (Drosophila), 22 groups and 1 complex were transferred to the subgenus Drosophila (Siphlodora) Patterson & Mainland and 6 groups, 2 species subgroups and 3 complexes are considered incertae sedis within the genus Drosophila. Different morphological characters provide different signals at different phylogenetic scales: thoracic characters (wing venation and presternal shape) discriminate families; grasping and erection-related characters discriminate subfamilies to tribes; whereas phallic paraphyses, i.e. auxiliary intromittent organs, discriminate genera and Drosophila subgenera. The study shows the necessity of analysing morphological characters within a molecular phylogenetic framework to translate molecular phylogenies into taxonomically-comprehensive classifications.

Phylogenetic relationships of 26 Phortica species were investigated based on DNA sequence data of two mitochondrial (ND2, COI) and one nuclear (28S rRNA) genes. Five monophyletic groups were recovered in the genus Phortica, of which three were established as new subgenera, Alloparadisa, Ashima, and Shangrila. The subgenus Allophortica was suggested as the most basal lineage in Phortica, followed by the lineage of P. helva + P. sobodo + P. varipes. The remaining Phortica species, most of Oriental distribution, formed a monophyletic group, and were subdivided into three lineages (i.e., the subgenera Ashima, Phortica, and Shangrila). The subgenera Shangrila and Phortica were suggested as sister taxa, and four clades were recovered in the subgenus Ashima. The result of reconstruction of ancestral distribution and estimation of divergence times indicates that, the ancestor of the genus Phortica restricted to Africa, its initial diversification was dated back to ca. 23 Mya (coinciding with the Oligocene/Miocene boundary); sympatric speciation and an Africa-to-Asia dispersal was proposed to account for the current distribution of Allophortica and the rest Phortica; most of the rest diversification of Phortica occurred in southern China, and the divergence between the African clade and its Oriental counterpart was suggested as a result of vicariance following a dispersal of their ancestral species from southern China to Africa.

The Stegana castanea species group is established as a morphological group within the subgenus Steganina, based on two known and six new species from the Oriental region: Stegana (Steganina) bacilla Chen and Aotsuka, 2004; S. castanea Okada, 1988; S. (S.) reni sp. nov.; S. (S.) tiani sp. nov.; S. (S.) tongi sp. nov.; S. (S.) wangi sp. nov., S. (S.) weiqiuzhangi sp. nov. and S. (S.) xui sp. nov. from southern China. A key to all the species of this group is provided.

The Phortica (Phortica) hani species complex is established, with descriptions of six new species discovered from the Hengduan to the Qinling mountains, China: Phortica floccipes Cao & Chen, Phortica hirtotibia Cao & Chen, Phortica longicauda Cao & Chen, Phortica longiseta Cao & Chen, Phortica panda Cao & Chen, and Phortica pinguiseta Cao & Chen spp. nov. A key to all the species of the hani complex is provided based on morphological data. In addition, a ‘molecular’ key to seven species of this complex was also elaborated based on DNA sequences of the mitochondrial ND2 and COI genes. The relationships amongst these seven species, and their relationships to the Phortica magna and Phortica variegata species complexes, are analysed based on the ND2 and COI sequence data. The results suggest that the P. hani complex is monophyletic; the P. magna and P. variegata complexes are closer to each other than either is to the P. hani complex, and both are closer to the P. hani complex than the Phortica foliiseta species complex or the Phortica varipes species group.

This paper reports on nine Asian species of the genus Paraleucophenga, of which four are new to science: Paraleucophenga brevipenis sp. nov., Paraleucophenga hirtipenis sp. nov., Paraleucophenga longiseta sp. nov., and Paraleucophenga tanydactylia sp. nov. We also report on a new synonym, Paraleucophenga shanyinensis Chen & Toda, 1994 syn. nov. A key to all of the species examined, based on morphological data, is provided, together with a ‘molecular’ key to seven Paraleudophenga species based on DNA sequence data of the mitochondrial NADH dehydrogenase subunit 2 (ND2) gene. The phylogenetic relationships among seven Paraleucophenga species are reconstructed based on DNA sequences of the mitochondrial ND2 gene, using two Leucophenga species as outgroups. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 155, 615–629.

[Extract] Flies ranked within the subfamily Steganinae (Diptera, Drosophilidae) often display a zoophilic parasitic behaviour both at larval and/or adult stages (Bachli et al. 2005; Otranto et al. 2008). Although drosophilid flies are commonly known as "fruit flies" as they usually feed and grow on fruit and other vegetable matter, some species have adapted to feed on vertebrate lachrymal secretions, potentially acting as intermediate hosts of Thelazia (Spirurida, Thelaziidae) spp. eyeworms (reviewed by Otranto et al. 2008). Phortica variegata Fallen 1823 has been proven to act as the intermediate host and vector of Thelazia callipaeda that infects the eyes of humans, rabbits and domestic and wild carnivores in Europe (Maca 1977; Otranto et al. 2005, 2006a,b, 2007). Despite recent reports suggesting that P. variegata acts as the main vector for T. callipaeda in Europe, a closely related species, namely Phortica semivirgo Maca, was previously suggested to play a role in the transmission of the nematode in eastern European countries (Maca 1977).

Three new species of the subgenus Parapenthecia of the genus Apenthecia are reported from the Oriental Region: A. (P.) heterochaeta Chen and Toda, sp. nov. from Indonesia, and A. (P.) hispida Chen and Toda, sp. nov. and A. (P.) litongi Cao and Chen, sp. nov. from south China. Interestingly, the last species lacks a very peculiar character, the swollen or scale-like orbital setae, which are commonly seen in all other species of the genus Apenthecia. A key to all species of the subgenus Parapenthecia is provided.

The subfamily Steganinae (Diptera, Drosophilidae) includes flies which display zoophilic feeding behaviour in the larval and/or adult stages, some of which act as vectors of Spirurida eyeworms, which infect both carnivores and humans. To date, the taxonomy and phylogeny of the subfamily Steganinae has been studied only superficially and many aspects of their systematics remain unresolved. Thus, the present study aimed to provide a molecular dataset to facilitate the identification and phylogenetic analysis of Steganinae species based on partial ( approximately 700 basepairs) mitochondrial cytochrome c oxidase subunit 1 (cox1) sequences. A total of 134 flies belonging to 13 species and eight genera of Steganinae were subjected to molecular and phylogenetic analyses. The mean nucleotide variation within the Steganinae subfamily was 8.1%, with a variation within genera for which more than one species was examined ranging from 1.6% (in Phortica spp.) to 21.8% (in Amiota spp.). Interspecific pairwise divergence ranged from 1.6% (Phortica variegata vs. Phortica semivirgo) to 24.8% (Cacoxenus indagator vs. Amiota alboguttata) and intraspecific variation ranged from 0% to 1%. Seventy of the 233 amino acids were variable, including 26 parsimony informative sites and 44 singleton sites, with some highly conserved residues identified within the genera Stegana and Amiota. Parsimony and maximum likelihood-based phylogenetic analyses provided strong support for the genus Phortica, phylogenetically distinct from the genus Amiota. Gitona distigma was placed in an unresolved position adjacent to the outgroup taxa, Drosophila yakuba and Drosophila melanogaster. The molecular data reported here represent the first molecular dataset based on cox1 of Steganinae flies and provide a base for further investigations into the evolutionary relationships among this little-studied subfamily.

Two new species of the subgenus Cacoxenus (Nankangomyia) Maca and Lin, 1993 are described: C. (N.) gani sp. nov. from southwest China and C. (N.) unipaxillus sp. nov. from Myanmar. The diagnosis of Nankangomyia is revised.

Thelazia callipaeda (Spirurida, Thelaziidae) is a small nematode living in the conjunctival sac of domestic and wild carnivores, rabbits and humans causing lacrimation, epiphora, conjunctivitis, keratitis and even corneal ulcers. The first autochthonous cases of thelaziosis affecting four dogs and one cat living in South Western France (Dordogne area) are reported and described. Nematodes recovered from the animals were morphologically identified as T. callipaeda and a partial region of the cytochrome oxidase c subunit 1 gene (cox1) was amplified by PCR from nematode specimens (from two dogs and the cat). In each case, this was shown to have an identical sequence to the haplotype 1 (h1) of T. callipaeda. So far, the arthropod acting as intermediate host of T. callipaeda eyeworms has not been identified in France although it might be Phortica variegata (Steganinae, Drosophilidae) as recently described in Italy.