Austrocnephia, new genus, for five species of 'Paracnephia' (Diptera: Simuliidae), with a key to Australian black fly genera.

A segregate of the so-called Australian 'Paracnephia' (Diptera: Simuliidae) is assigned to a new genus, Austrocnephia. The taxon is fully diagnosed and a key to constituent species presented. Two species-groups are recognized: the aurantiaca species-group, comprised of A. aurantiaca (Tonnoir 1925) and A. strenua (Mackerras Mackerras 1950), and the tonnoiri species-group, comprised of A. fuscoflava (Mackerras Mackerras 1948), A. orientalis (Mackerras Mackerras 1950) and A. tonnoiri (Drummond 1931). Both species-groups are diagnosed and the included species fully redescribed. Detailed locality data is given, as is information about biology, when known. Brief comments are offered about the historical biogeography of Austrocnephia. A key to Australian simuliid genera is also provided.

Diptera of Canada.

The Canadian Diptera fauna is updated. Numbers of species currently known from Canada, total Barcode Index Numbers (BINs), and estimated numbers of undescribed or unrecorded species are provided for each family. An overview of recent changes in the systematics and Canadian faunistics of major groups is provided as well as some general information on biology and life history. A total of 116 families and 9620 described species of Canadian Diptera are reported, representing more than a 36% increase in species numbers since the last comparable assessment by JF McAlpine et al. (1979). Almost 30,000 BINs have so far been obtained from flies in Canada. Estimates of additional number of species remaining to be documented in the country range from 5200 to 20,400.

A new genus, Protaustrosimulium, for four species of Australian black flies (Diptera: Simuliidae).

Protaustrosimulium n. gen. is described for four species: two previously named species from southeastern Australia-Paracnephia pilfreyi (Davies Györkös 1988) and Paracnephia terebrans (Tonnoir 1925)-plus two newly described ones from the southwestern-most corner of Western Australia-Prot. amphorum n. sp. and Prot. opscurum n. sp. Molecular and morphological data suggest a close relationship between members of the new genus and Austrosimulium Tonnoir 1925. Monophyly of Protaustrosimulium is supported mainly by characters of adult females, as two of the four species are known only in that life stage. Two species groups are recognized: the pilfreyi-group for Prot. pilfreyi and Prot. amphorum, and the terebrans-group for Prot. terebrans and Prot. opscurum. The constituent species in each group are distributed vicariously in southeastern and southwestern Australia-a common biogeographical pattern in Australian simuliids.

Nothogreniera new genus, for two species of Australian "Paracnephia" (Diptera: Simuliidae).

Two species of Australian Simuliidae known only from adult females and currently assigned to "Paracnephia" are re-described, as are their now-known males and immature stages. Morphological character states of "Paracnephia" fergusoni (Tonnoir) and "P." fergusoni var. (Mackerras Mackerras) reveal that they are markedly distinct from all other Australian species, and are here assigned to the new genus-Nothogreniera-the most plesiomorphic Gondwanan Australian simuliid. Structural variation among populations of N. fergusoni suggests that this entity comprises a species complex.

Precipitation and ectoparasitism reduce reproductive success in an arctic-nesting top-predator.

Indirect impacts of climate change, mediated by new species interactions (including pathogens or parasites) will likely be key drivers of biodiversity reorganization. In addition, direct effects of extreme weather events remain understudied. Simultaneous investigation of the significance of ectoparasites on host populations and extreme weather events is lacking, especially in the Arctic. Here we document the consequences of recent black fly outbreaks and extreme precipitation events on the reproductive output of an arctic top predator, the peregrine falcon (Falco peregrinus tundrius) nesting at the northern range limit of ornithophilic black flies in Nunavut, Canada. Overall, black fly outbreaks and heavy rain reduced annual nestling survival by up to 30% and 50% respectively. High mortality caused by ectoparasites followed record-breaking spring snow precipitation, which likely increased stream discharge and nutrient runoff, two key parameters involved in growth and survival of black fly larvae. Using the RCP4.5 intermediate climate scenario run under the Canadian Global Climate Model, we anticipate a northward expansion of black fly distribution in Arctic regions. Our case study demonstrates that, in the context of climate change, extreme weather events can have substantial direct and indirect effects on reproductive output of an arctic top-predator population.

Reassignment of Western Australia Paracnephia gladiator Moulton Adler to a new genus, Bunyipellum (Diptera: Simuliidae).

With new material available of most stages of many known Australian Paracnephia, including new species, it is now clear that certain segregates warrant assignment to new genera. This applies to Paracnephia gladiator Moulton Adler, a Western Australia simuliid with numerous unique character states. The species is fully redescribed and assigned to Bunyipellum nov. gen. A diagnosis is provided and relationships discussed, as is historical biogeography. Bunyipellum appears to be more closely related to elements of the South American simuliid fauna than to any other Gondwanan Australian species.

Taxonomic revision of Paraustrosimulium Wygodzinsky & Coscarón: reassignment of Austrosimulium colboi and description of P. obcidens n. sp. from Western Australia.

The hitherto monotypic South American genus Paraustrosimulium Wygodzinsky & Coscarón is revised to accommodate two Australian species: Austrosimulium colboi Davies & Györkös and Paraustrosimulium obcidens n. sp. The generic diagnosis is updated and the eastern Australian species Paraustrosimulium colboi (Davies & Györkös) n. stat. is re-described, including the male for the first time. The Western, Australian sister species of P. colbo, namely P. obcidens Craig, Moulton Currie n. sp. is also fully described. The relationship of Paraustrosimulium to other simuliid genera is discussed, as are aspects of historical biogeography.

The speciation continuum: population structure, gene flow, and maternal ancestry in the Simulium arcticum complex (Diptera: Simuliidae).

Comparative analyses of populations at different stages of divergence can yield insights into the process of speciation. We assess population structure, gene flow, and maternal ancestry at five locations containing sympatric members of the Simulium arcticum complex at different stages of chromosome divergence. We analyze both nuclear and mitochondrial DNA markers, including 11 microsatellite loci, as well as COI, COII, cytb, and ND4 gene sequences. Simulium negativum, representing the later stages of divergence, shows both nuclear and mitochondrial differentiation when compared with allopatric and sympatric chromosomal forms, as well as both low contemporary and historical gene flow in sympatry. At intermediate stages of chromosome divergence, populations differ at nuclear, but not mitochondrial, loci in allopatry and sympatry. In one comparison of intermediate stage chromosomal forms (S. arcticum sensu stricto and S. apricarium), populations demonstrate low contemporary, but higher historical, gene flow in sympatry. In a second sympatric comparison (S. arcticum s. s. and S. brevicercum), both contemporary and historical gene flow are high. All analyses of sympatric populations at the earliest stages of chromosome divergence demonstrate panmixia; yet, some nuclear differentiation in allopatry is apparent. These findings suggest that molecular divergence is tracking chromosome divergence along a chromosomally-defined continuum of speciation in black flies.

Good species behaving badly: Non-monophyly of black fly sibling species in the Simulium arcticum complex (Diptera: Simuliidae).

Mitochondrial based phylogenetic reconstructions often show deviations from species-level monophyly. We used the Simulium arcticum species complex (Diptera: Simuliidae) as a model system for interpreting non-monophyly in light of chromosomal data supporting species status of siblings. For cytogenetic identification of morphologically indistinguishable black fly sibling species, larvae must be preserved in Carnoy's solution, a fixative known to degrade DNA. Consequently, we reconstructed phylogenetic relationships based on 12S, COII, cyt b, and ITS-1 gene sequences obtained from larvae sampled from presumed taxon-pure localities. As species composition at 'taxon-pure' sites may have changed at the time of sampling, we performed a second study that aimed to: (1) assess phylogenetic relationships among cytologically verified members of the S. arcticum species complex using COI and COII gene sequences; (2) determine whether useable genetic information could be gleaned from Carnoy's fixed specimens; and (3) determine the extent to which Carnoy's fixative degrades DNA over time. We consistently obtained genetic data from material stored in Carnoy's solution for two to three months. Genetic analysis of samples fixed in Carnoy's solution for up to six years indicates that larvae preserved for a maximum of five years can provide useable information for molecular analysis. Our preliminary and cytologically confirmed phylogenetic analyses demonstrate that mitochondrial DNA fails to resolve species-level monophyly of chromosomally distinct S. arcticum taxa. As results of analyses based on cytologically verified larvae mirror those of our preliminary study, we rule out imperfect taxonomy as the reason for species-level non-monophyly. Although we cannot confidently reject either inadequate phylogenetic information or incomplete lineage sorting as the cause of non-monophyly, the sharing of alleles between sympatric siblings suggests introgressive hybridization between taxa. We conclude that the patterns present in the S. arcticum phylogeny likely represent the initial stages of chromosome based sibling speciation.

Simulium (Inaequalium) marins, a new species of black fly (Diptera: Simuliidae) from inselbergs in Brazil.

A new species of black fly, Simulium (Inaequalium) marins (Diptera: Simuliidae), is described based on the male, female, pupa and larva. This new species was collected from two localities: a small stream on the Pico dos Marins, a high mountain with granite outcrops in Piquete County, state of São Paulo, and in a small stream in the Serra dos Orgãos National Park, state of Rio de Janeiro, Brazil.

Simulium (Inaequalium) marins, a new species of black fly (Diptera: Simuliidae) from inselbergs in Brazil

A new species of black fly, Simulium (Inaequalium) marins (Diptera: Simuliidae), is described based on the male, female, pupa and larva. This new species was collected from two localities: a small stream on the Pico dos Marins, a high mountain with granite outcrops in Piquete County, state of São Paulo, and in a small stream in the Serra dos Órgãos National Park, state of Rio de Janeiro, Brazil.

Isolation and characterization of 11 microsatellite loci from the black fly, Simulium negativum (Diptera: Simuliidae).

Eleven polymorphic microsatellite loci were isolated and developed from the black fly, Simulium negativum, a member of the Simulium arcticum sibling species complex. The observed heterozygosity of the 11 loci ranged from 0.03 to 0.83. The number of alleles per locus ranged from 8 to 19. Significant linkage disequilibria were encountered only for the primer pairs BF7-1 with BF7-5 and BF6-32 with BF7-16. Presumably, these microsatellite loci can be used to study genetic structure within the entire S. arcticum complex.

Identification of Nearctic black flies using DNA barcodes (Diptera: Simuliidae).

DNA barcoding has gained increased recognition as a molecular tool for species identification in various groups of organisms. In this preliminary study, we tested the efficacy of a 615-bp fragment of the cytochrome c oxidase I (COI) as a DNA barcode in the medically important family Simuliidae, or black flies. A total of 65 (25%) morphologically distinct species and sibling species in species complexes of the 255 recognized Nearctic black fly species were used to create a preliminary barcode profile for the family. Genetic divergence among congeners averaged 14.93% (range 2.83-15.33%), whereas intraspecific genetic divergence between morphologically distinct species averaged 0.72% (range 0-3.84%). DNA barcodes correctly identified nearly 100% of the morphologically distinct species (87% of the total sampled taxa), whereas in species complexes (13% of the sampled taxa) maximum values of divergence were comparatively higher (max. 4.58-6.5%), indicating cryptic diversity. The existence of sibling species in Prosimulium travisi and P. neomacropyga was also demonstrated, thus confirming previous cytological evidence about the existence of such cryptic diversity in these two taxa. We conclude that DNA barcoding is an effective method for species identification and discovery of cryptic diversity in black flies.