Simulium (Gomphostilbia) fukudae sp. nov. is described based on females, males, pupae, and larvae from Thailand. This new species is placed in the Simulium batoense species-group of Simulium (Gomphostilbia). It is characterized by the female frons almost bare, male paramere covered with minute setae, pupal gill composed of six slender filaments, pupal terminal hooks cone-shaped, and cocoon with an anterodorsal projection. Taxonomic notes are given to separate this new species from the most similar species, Simulium (G.) sextuplum Takaoka & Davies and Simulium (G.) paukatense Takaoka, both from Malaysia.
Simulium (Gomphostilbia) maleewongae sp. nov. is described based on the adult males and females, their pupal exuviae, and larvae from Thailand. This new species is placed in the Simulium gombakense species-group of Simulium (Gomphostilbia). It is characterized by the female cibarium with a cup-like appendage, male ventral plate deeply depressed ventromedially, pupal gill composed of an inflated structure and eight slender filaments, cone-shaped pupal terminal hooks, and cocoon with an anterodorsal projection. Taxonomic notes are given to separate this new species from 10 other species of the same species-group known from China, India, Malaysia, Thailand, Nepal, and Vietnam. Keys to identify all 11 species of the S. gombakense species-group are provided for females, males, pupae, and larvae.
A faunistic survey of black flies in Shan State, central Myanmar, was carried out in 2013. A total of 16 species were collected, of which 13 species are newly recorded from Myanmar. Among 13 newly recorded species, S. (S.) chiangmaiense Takaoka & Suzuki varied in the number of pupal gill filaments from eight to 10. This survey increases the number of species of black flies from Myanmar from 8 to 23. They are classified in five subgenera of the genus Simulium: one in Asiosimulium, seven in Gomphostilbia, one in Montisimulium, two in Nevermannia and 12 in Simulium.
Simulium (Asiosimulium) saeungae sp. nov. (Diptera: Simuliidae) is described based on females, males, pupae, and mature larvae collected from Nan Province, Northern Thailand. It is characterized by the medium-long cerci in the female, enlarged hind basitarsus, and broad ventral plate with its posterior margin not deeply concave in the male, arborescent pupal gill with 42-56 filaments in the pupa and smaller number of primary rays of the labral fan (30-33) in the larva. This is the fifth species of the subgenus Asiosimulium, the second smallest among 10 subgenera in the Oriental Region. Taxonomic notes are given to distinguish this new species from the three known species from Thailand and one from Nepal.
Five new species of black flies, Simulium (Gomphostilbia) chiangraiense, S. (G.) huaikaeoense, S. (G.) khaokhoense, S. (G.) maeklangense, and S. (G.) pamiangense, are described based on adults, pupae and mature larvae from Thailand. These five species are similar to one another and are placed in the S. decuplum subgroup of the S. batoense species-group by having the male fore coxae darkened, ventral plate flat and transverse; pupal gill with 10 short filaments; and larval postgenal cleft deep approaching or reaching the posterior margin of the hypostoma. Simulium (G.) chiangraiense sp. nov. and S. (G.) pamiangense sp. nov. differ from the three other new species by the much longer pupal gill filaments and darkened dorsal surface of abdominal segments 1 and 2. Taxonomic notes are provided to distinguish these five new species from five other related species. The phylogenetic positions of these new species in the S. decuplum subgroup are presented based on the mitochondrial COI gene. Simulium (G.) pamiangense sp. nov. and S. (G.) huaikaeoense sp. nov. are highly similar to each other genetically, showing a sister relationship, though they are clearly different morphologically. On the contrary, S. (G.) khaokhoense sp. nov. is distantly positioned from S. (G.) huaikaeoense nov., though it is almost indistinguishable morphologically from the latter.
The female, male, pupa and mature larva of Simulium (Simulium) poolpholi sp. nov. collected from a large and seasonal stream in northeastern Thailand are described. This new species is placed in the S. striatum species-group by the female sternite 8 concave posteromedially in form of the inverted-U shape, ovipositor valve with a ventrally produced process along its inner margin, male ventral plate saddle-shaped, style with a basal protuberance, and pupal gill with 10 filaments. It is morphologically similar to S. (S.) phraense Takaoka, Srisuka & Saeung from northern Thailand by lacking annular ridges on the surface of pupal gill filaments but it is distinguished from the latter species by the arrangement of pupal gill filaments. Molecular analysis using the fast-evolving nuclear big zinc finger (BZF) gene shows that S. (S.) poolpholi sp. nov. formed a distinct clade, being separated from the seven other Thai species of this species-group by a genetic distance of 1.84-4.55%. Our discovery increases the number of species in the S. striatum species-group in Thailand to nine.
A new black fly species, Simulium (Gomphostilbia) wijiti, is described based on adult females, males, pupal exuviae and mature larvae from Mae Hong Son Province, Thailand. This new species is placed in the Simulium ceylonicum species-group. It is distinguished from four Thai members of the S. ceylonicum species-group [S. (G.) curtatum Jitklang et al., S. (G.) pangsidaense Takaoka, Srisuka & Saeung, S. (G.) sheilae Takaoka & Davies, and S. (G.) trangense Jitklang et al.], in the female by the short to medium long sensory vesicle; in the male by the large number of upper-eye (large) facets in 15 vertical columns and 15 or 16 horizontal rows; in the pupa by the dorsum of abdominal segments darkened; and in the larva by the antenna as long as or slightly shorter than the stem of the labral fan (longer than the stem of the labral fan in four other species). Phylogenetic analysis based on the COI gene sequences revealed that this new species is genetically closely related to S. leparense of the S. ceylonicum species-group, but is clearly separated from the latter species, and also from the three Thai related species (S. curtatum, S. sheilae and S. trangense) of the same species-group with interspecific genetic distances ranging from 9.65% to 12.67%. This is the fifth member of the S. ceylonicum species-group recorded from Thailand.
Females and males reared from pupae, their pupal exuviae and cocoons, and mature larvae of the Simulium (Gomphostilbia) asakoae species group from various localities in Thailand were morphologically examined. A total of 25 species was identified, including two of four known species (Simulium asakoae Takaoka & Davies and S. chiangdaoense Takaoka & Srisuka), one newly confirmed species (S. myanmarense Takaoka, Srisuka & Saeung, originally described from Myanmar), one newly transferred species (S. inthanonense Takaoka & Suzuki formerly of the S. ceylonicum species group), and 21 new species. Descriptions of all 21 new species are given, and the first full description of the male of S. inthanonense, together with the revised descriptions of its female, pupa, and larva, is also provided. Keys to identify all 27 members of this species group from Thailand are given for females, males, pupae, and larvae. The genetic relationships of all but one species were resolved using COI gene sequence-based analysis. All 26 species were divided into nine subgroups, I-IX, each consisting of two, one, four, nine, one, three, two, one and three species, respectively.
Simulium (Gomphostilbia) leparense sp. nov. is described from females, males, and pupae collected from Peninsular Malaysia. This new species is assigned to the ceylonicum species-group of the subgenus Gomphostilbia, and is characterized by the female and male scuta covered with dark-brown short hairs, smaller number of male upper-eye facets, presence of shiny paired spots on the male abdominal segments 2-8, and absence of grapnel-shaped hooklets on the pupal abdominal segment 9. The male and pupa of S. capillatum Takaoka, which was originally described from larvae collected from Sarawak and Sabah, are described for the first time. Keys to identify all 32 species of the Simulium ceylonicum species-group including 27 species from other countries are provided for females, males, pupae, and mature larvae.
Two new species of black flies, Simulium (Comphostilbia) terengganuense sp. nov. and Simulium (Gomphostilbia) aziruni sp. nov. (Diptera: Simuliidae), are described on the basis of reared adult, pupal, and larval specimens collected from Peninsular Malaysia. Both species are placed in the batoense species-group within the subgenus Gomphostilbia, one of two dominant subgenera of the genus Simulium in Peninsular Malaysia as well as in the Oriental Region. Strikingly, three morphological characteristics that rarely occur in the subgenus Gomphostilbia are found in these two new species: the very narrow female frons and the mushroom-like pupal terminal hooks in S. (G.) terengganuense sp. nov. and the pupal gill composed of an inflated horn-like structure and eight slender filaments in S. (G.) aziruni sp. nov.
Hard ticks taken from the Japanese serow, Capricornis crispus, in Yamagata Prefecture, Honshu, harboured infective larvae of onchocercid filariae after incubation from the 22nd to the 158th day. Haemaphysalis flava and H. japonica contained one to eight filarial larvae; females, males and a nymph of the ticks were infected. The 44 infective larvae recovered were 612-1,370 μm long, and 11 of them, 930-1,340 μm long, were studied in detail. The larvae possessed the morphologic characteristics of the larvae of the genus Cercopithifilaria, namely an oesophagus with a posterior glandular part, no buccal capsule and a long tail with three terminal lappets. Five types (A to E) of infective larvae were identified based on the morphologic characteristics. While to date five species of Cercopithifilaria have been described from the Japanese serow, a specific identification of the larvae found in this study was generally not possible. Only type E larvae could be tentatively assigned to Cercopithifilaria tumidicervicata, as they had a cervical swelling similar to that of the adults of this species. A key for the identification of the five larval types is presented. The study presents circumstantial evidences indicating that H. flava and H. japonica may transmit Cercopithifilaria spp. to Japanese serows. It also suggests the possibility that such filarial larvae will be found in hard ticks anywhere, because Cercopithifilaria is distributed worldwide, though this genus generally goes unnoticed, as its microfilariae occur in the skin, not in the blood, of host animals.
Simulium (Nevermannia) khunklangense sp. nov. is described from females, males, pupae and larvae collected in Doi Inthanon National Park, Chiang Mai, Thailand. This new species is placed in the vernum species-group of the subgenus Nevermannia and is similar to S. (N.) chomthongense Takaoka & Srisuka described from Doi Inthanon National Park, Thailand, but is distinguished in the male by the number of enlarged upper-eye facets and the relative width of the hind basitarsus against the hind tibia and femur, and in the pupa by the short common basal stalk of the gill and the cocoon with an anterodorsal bulge or a short anterodorsal projection. Taxonomic notes are provided to separate this new species from five other known species of the vernum species-group, which share an accessory sclerite on the larval abdomen, a rare characteristics in this species-group.
To investigate the breeding habitat preference of black flies, a comprehensive black fly survey was conducted for the first time in Peninsular Malaysia. Preimaginal black flies (pupae and larvae) were collected manually from 180 stream points encompassing northern, southern, central and east coast of the Peninsular Malaysia. A total of 47 black fly species were recorded in this study. The predominant species were Simulium trangense (36.7%) and Simulium angulistylum (33.3%). Relatively common species were Simulium cheongi (29.4%), Simulium tani (25.6%), Simulium nobile (16.2%), Simulium sheilae (14.5%) and Simulium bishopi (10.6%). Principal Component Analysis (PCA) of all stream variables revealed four PCs that accounted for 69.3% of the total intersite variance. Regression analysis revealed that high species richness is associated with larger, deeper, faster and higher discharge streams with larger streambed particles, more riparian vegetation and low pH (F=22.7, d.f.=1, 173; P<0.001). Relationship between species occurrence of seven common species (present in >10% of the sampling sites) was assessed. Forward logistic regression analysis indicated that four species were significantly related to the stream variables. S. nobile and S. tani prefer large, fast flowing streams with higher pH, large streambed particles and riparian trees. S. bishopi was commonly found at high elevation with cooler stream, low conductivity, higher conductivity and more riparian trees. In contrast, S. sheilae was negatively correlated with PC-2, thus, this species commonly found at low elevation, warmer stream with low conductivity and less riparian trees. The results of this study are consistent with previous studies from other geographic regions, which indicated that both physical and chemical stream conditions are the key factors for black fly ecology.
A new black fly species, Simulium (Gomphostilbia) pangsidaense, is described on the basis of adult female, male, pupal exuviae and mature larvae from Pang Sida National Park, Sa Kaew Province, Eastern Thailand. This new species is placed in the Simulium ceylonicum species-group. It is distinguished from three Thai members of the S. ceylonicum species-group by the following characteristics: from S. (G.) curtatum Jitklang et al. and S. (G.) sheilae Takaoka & Davies by the wide pupal terminal hooks (triangular terminal hooks in the latter two species), and from S. (G.) sheilae and S. (G.) trangense Jitklang et al. by the number of male upper-eye facets in 13 vertical columns and 14 or 15 horizontal rows (10 or 11 vertical columns and 12 or 13 horizontal rows in the latter two species). This is the fourth member of the S. ceylonicum species-group recorded from Thailand.
The female and larva of Simulium (Gomphostilbia) udomi Takaoka & Choochote from Thailand are described for the first time. The female of this species is similar to those of S. (G.) asakoae Takaoka & Davies from Peninsular Malaysia, Thailand, Hong Kong and Vietnam, and S. (G.) chiangdaoense Takaoka & Srisuka from Thailand. The larva of this species is similar to S. (G.) curtatum Jitklang et al. and S. (G.) nr. asakoae 2 from Thailand in having a medium-long postgenal cleft. Taxonomic notes are given to separate this species from these related species. The COI gene sequence of S. (G.) udomi is compared with those of eight species of the S. asakoae species-group and three species of the S. ceylonicum species-group. This species is transferred from the S. ceylonicum species-group to the S. asakoae species-group based on the adult female and male morphological characters, comparisons of the genetic distances and phylogenetic relationships inferred from the COI gene sequences.
Simulium (Simulium) hackeri Edwards, 1928 of the Simulium variegatum species-group from Malaysia was described initially based on the female specimen from Cameron Highlands, Pahang. In the present study, the pupa and larva of this species are described for the first time. Their morphological characters resemble those of the Simulium variegatum species-group by having six gill filaments per side, abdomen with dorsal spine-combs at least on segments 7 and 8, cocoon with wall-pocket shaped and with or without an anterodorsal projection. Postgenal cleft of the larva medium-sized, rarely small, ventral papillae small or absent. The DNA barcode of this species is also reported herein.
Larvae and adults reared from pupae of Simulium (Gomphostilbia) gombakense Takaoka & Davies from eight streams in five provinces of Thailand were genetically analyzed by using cytochrome c oxidase subunit 1 sequences. The material was composed of four lineages, of which lineages 2, 3 and 4 are genetically closely related to one another, but more distantly related to lineage 1, which is equivalent to typical S. (G.) gombakense from Peninsular Malaysia. Lineages 2, 3 and 4, which are morphologically indistinguishable, are designated as genoforms A, B and C within a species complex of S. (G.) paiense sp. nov., which is described based on specimens of lineage 2 (=genoform A). Simulium (G.) paiense sp. nov. is similar to S. (G.) gombakense from Peninsular Malaysia but appears to be barely distinguished by the relative length of the female fore basitarsus. Simulium (G.) thuathienense Takaoka & Sofian-Azirun is newly recorded from Thailand and its male is described for the first time. The female of S. (G.) gombakense is briefly described based on females reared from pupae collected from Peninsular Malaysia.
Two new species of Simulium (Gomphostilbia), S. (G.) myanmarense and S. (G.) monglaense, are described from females, males, pupae and larvae from Myanmar. The two new species are placed in the S. asakoae species-group, and are similar to each other in the female and male but distinguished in the pupa by the presence or absence of an anterodorsal projection of the cocoon, and in the larva by a unique pattern of colored markings on the abdomen. Taxonomic notes are given to separate these species from related species. The COI gene sequences of both species are compared with those of eight species of the S. asakoae species-group and three species of the S. ceylonicum species-group. Both new species are most closely related to each other, further supporting their morphological classification in the S. asakoae species-group.
Seven populations of Simulium parahiyangum Takaoka & Sigit (Diptera: Simuliidae), a geographically widespread nominal species of black fly in Southeast Asia, were morphologically and molecularly studied. Three morphoforms based on male and pupal morphological features, and two primary lineages based on the COI gene sequence analysis were recognized. Morphoform 1 (lineage 1) from Sarawak, Malaysia, is identified as S. parahiyangum sensu stricto and morphoform 2 (lineage 2) from Thailand and Vietnam, and morphoform 3 (lineage 1) from Peninsular Malaysia are each regarded as distinct species, although morphoform 3 is partially homosequential for the COI gene with morphoform 1. Morphoforms 2 and 3 are described as Simulium ngaoense sp. nov. and Simulium sazalyi sp. nov., respectively. Overall, S. parahiyangum is not a single geographic generalist but a composite of multiple species.
A black fly species of the Simulium feuerborni species-group of Simulium (Nevermannia) from Cameron Highland, Peninsular Malaysia, previously regarded as S. feuerborni Edwards, originally described from East Java, is described as Simulium pairoti sp. nov. based on complete life stages. High intraspecific variations in the arrangement of the six pupal gill filaments, length of the stalk of the ventral paired filaments, and length of the anterodorsal projection of the cocoon, are noted in this species. This new species is readily distinguished from its congeners by having the characters of male genitalia with simple lamellate ventral plate, short inwardly-twisted styles, several parameral hooks, and a simple narrow median sclerite. Morphological data reported herein plus the chromosomal and molecular data presented elsewhere support S. pairoti as a novel pseudocryptic species.