Biting midges of genusCulicoides(Diptera: Ceratopogonidae) are the vectors of several pathogenic arboviruses and parasites of humans and animals. Several reports have suggested that biting midges might be a potential vector ofLeishmaniaparasites. In this study, we screened forLeishmaniaandTrypanosomaDNA in biting midges collected from near the home of a leishmaniasis patient in Lamphun province, northern Thailand by using UV-CDC light traps. The identification of biting midge species was based on morphological characters and confirmed using the Cytochrome C oxidase subunit I (COI) gene. The detection ofLeishmaniaandTrypanosomaDNA was performed by amplifying the internal transcribed spacer 1 (ITS1) and small subunit ribosomal RNA (SSU rRNA) genes, respectively. All the amplified PCR amplicons were cloned and sequenced. The collected 223 biting midges belonged to seven species (Culicoides mahasarakhamense,C.guttifer,C.innoxius,C.sumatrae,C.huffi,C.oxystoma, andC.palpifer). The dominant species found in this study wasC.mahasarakhamense(47.53%).Leishmania martiniquensisDNA was detected in three samples of 106 specimens ofC.mahasarakhamensetested indicating a field infection rate of 2.83%, which is comparable to reported rates in local phlebotomines. Moreover, we also detectedTrypanosomasp. DNA in one sample ofC.huffi. To our knowledge, this is the first molecular detection ofL.martiniquensisinC.mahasarakhamenseas well as the first detection of avianTrypanosomainC.huffi. Blood meal analysis of engorged specimens ofC.mahasarakhamense,C.guttifer, andC.huffirevealed that all specimens had fed on avian, however, further studies of the host ranges ofCulicoidesare needed to gain a better insight of potential vectors of emerging leishmaniasis. Clarification of the vectors of these parasites is also important to provide tools to establish effective disease prevention and control programs in Thailand.

An entomological survey at rural and cavernicolous localities in four provinces in southern Thailand provided 155 blood-fed females of sand flies (Diptera: Psychodidae) that were identified based on morphological characters as Idiophlebotomus asperulus (n = 19), Phlebotomus stantoni (n = 4), P. argentipes (n = 3), Sergentomyia anodontis (n = 20), S. barraudi (n = 9), S. hamidi (n = 23), S. hodgsoni (n = 4), S. hodgsoni hodgsoni (n = 32), S. indica (n = 5), S. iyengari (n = 2), S. khawi (n = 17), S. silvatica (n = 11) and Sergentomyia sp. (n = 6). The dominant species in this study was S. hodgsoni hodgsoni, which was collected specifically in a Buddha cave. Screening for DNA of parasitic protozoans revealed eight specimens (5.16%) of four species (S. barraudi, S. indica, S. khawi and Id. asperulus) positive for Trypanosoma sp., while no Leishmania spp. DNA was detected. Blood meals of engorged females were identified by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) assay on a fragment of cytochrome b (cyt b) gene with a success rate 36%, humans, dogs, and rats being determined as sources of blood. Bloodmeal analysis of two Trypanopsoma-positive females (S. barraudi and Sergentomyia sp.) identified blood from dogs and humans, respectively. Our findings indicate that S. barraudi, S. indica, S. khawi and Id. asperulus may be incriminated in circulation of detected Trypanosoma spp.

Recent focus on Culicoides species diversity in Thailand was prompted by a need to identify vectors responsible for the transmission of African Horse Sickness in that country. To assist rapid genetic identification of species, we sampled mitochondrial cytochrome c oxidase subunit I (COI) DNA barcodes (N = 78) from 40 species of Culicoides biting midge from Thailand, including 17 species for which DNA barcodes were previously unavailable. The DNA barcodes were assigned to 39 Barcode Identification Numbers (BINs) representing terminal genetic clusters at the Barcode of Life Data systems (BOLD). BINs assisted with comparisons to published conspecific DNA barcodes and allowed partial barcodes obtained from seven specimens to be associated with BINs by their similarity. Some taxonomic issues were revealed and attributed to the possible misidentification of earlier reported specimens as well as a potential synonymy of C. elbeli Wirth & Hubert and C. menglaensis Chu & Liu. Comparison with published BINs also revealed genetic evidence of divergent population processes and or potentially cryptic species in 16 described taxa, flagged by their high levels of COI sequence difference among conspecifics. We recommend the BOLD BIN system to researchers preparing DNA barcodes of vouchered species for public release. This will alert them to taxonomic incongruencies between their records and publicly released DNA barcodes, and also flag genetically deep and potentially novel diversity in described species.

Culicoides biting midges were collected using UV light traps from nine livestock farms in Kanchanaburi Province, Thailand. Collections were made one night per month from June 2020 to May 2021 to determine the seasonal changes and spatial distribution of the Culicoides assemblage. The influence of four environmental factors (temperature, rainfall, humidity, and wind speed), types of animals in each shed (cattle, pigs, and avians), and neighboring animals (those in the other sheds and their proximity) were assessed. A subsample of 130,670 out of a total of 224,153 specimens collected was identified and counted. The collections were predominantly female (76.9%), though males were also commonly collected (23.1%). The dominant species included C. peregrinus (97,098 individuals; 43.0%), C. oxystoma (55,579; 24.6%), C. arakawae (45,996; 20.4%), and C. imicola (15,703; 7.0%), while other species accounted for the remaining 9777 individuals (4.4%). Trap captures were strongly influenced by seasons and animal sheds. Cattle exhibited the greatest effect on the community, likely due to their large biomass. Humidity and temperature had a positive effect on trap captures, whereas wind speed exhibited a negative effect. Cattle positively influenced all major species, except for C. arakawae, which showed a positive association with avians. A “neighboring effect” was also observed. Additionally, the lowest dispersal ability of C. arakawae was suggested.