Two new black-fly species of the subgenus Simulium (Simulium) are described and illustrated, based on female specimens captured while approaching a human near a mountain summit in Doi Inthanon National Park, Chiang Mai Province, northern Thailand. The first species, Simulium suchariti sp. nov., is assigned to the griseifrons species-group and is distinguished from most other related species by the cibarium with tubercles near the base of its medial projection; the second, S. setsukoae sp. nov., is very similar to S. rufibasis of the tuberosum species-group but is distinguished from the latter by the shorter length of clustered stout hairs on the seventh abdominal segment and the ovipositor valve with its posteromedial corner widely bare.

Forty-five known species of Simulium Latreille s. l. in Thailand are listed, and keys to subgenera and species within each subgenus are provided for adults, pupae and mature larvae.

Two known and two new species of the griseifrons species-group of the subgenus Simulium (Simulium) in Northern Thailand are reported. The male, pupa and mature larva of S. (S.) rudnicki Takaoka and Davies, and the pupa and mature larva of S. (S.) suchariti Takaoka and Choochote, are described for the first time. Furthermore, two new simuliid species, S. (S.) mediocoloratum sp. nov. and S. (S.) crocinum sp. nov., both of which are very similar to S. (S.) rudnicki, are also described.

Simulium (Simulium) phukaense sp. nov. is described on the basis of the observation of females and males (both sexes of adults reared from pupae) and pupae collected in Nan Province, northern Thailand. This new species is assigned to the griseifrons species-group of the subgenus Simulium (Simulium) and is easily distinguished from other known species of this species-group by the simple shoe-shaped cocoon, as well as the arrangement of the six gill filaments.

Two new species of black flies, Simulium (Nevermannia) fruticosum sp. nov. and S. (N.) chiangklangense sp. nov. are described on the basis of samples collected in northern Thailand. These new species are assigned to the feuerborni species-group of the subgenus Simulium (Nevermannia), and are easily distinguished from S. (N.) feuerborni Edwards, the only species of the same group known thus far in Thailand, by their simple cocoon without any anterodorsal projection.

Two more new black-fly species of the rare subgenus Simulium (Montisimulium) were discovered in Doi Inthanon National Park, Chiang Mai, Thailand, where S. (M.) merga Takaoka and Choochote was known as the only named species. These two new species, S. (M.) angkaense sp. nov. and S. (M.) laoleense sp. nov., are described on the basis of the pupal and⁄or mature larvae. Both new species are easily distinguished from S. (M.) merga by the pupal gill with 12 slender filaments, and from all the 16 known species with 12 pupal gill filaments in other countries by the long and very long common basal stalk of the gill, respectively.

Simulium (Simulium) chanyae sp. nov. is described based on two female adult specimens collected in Northern Thailand. This new species is assigned to the multistriatum species-group of the subgenus Simulium (Simulium), and is easily distinguished from most of the other known species of this species-group by the yellowish legs.

A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5+/-0.2degrees C, 79.0+/-0.3degrees C, 76.8+/-0.1degrees C, and 79.9+/-0.1degrees C, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.
Animals ; Blood/*parasitology ; Brugia/classification/genetics/*isolation & purification ; Cats ; Culicidae/*parasitology ; Dirofilaria immitis/classification/genetics/*isolation & purification ; Dogs ; Humans ; Male ; Parasitology/*methods ; RNA, Helminth/genetics ; RNA, Ribosomal, 5S/genetics ; Real-Time Polymerase Chain Reaction/*methods ; Sensitivity and Specificity ; Transition Temperature ; Wuchereria bancrofti/classification/genetics/*isolation & purification