METHODS: We isolated Onchocerca specimens from bearded pigs and examined their morphology. For comparative material, we collected fresh specimens of O. d. dewittei Bain, Ramachandran, Petter & Mak, 1977 from banded pigs (S. scrofa vittatus Boie) in Peninsular Malaysia. Partial sequences of three different genes (two mitochondrial genes, cox1 and 12S rRNA, and one nuclear ITS region) of these filarioids were analysed. By multi-locus sequence analyses based on six genes (16S rDNA, ftsZ, dnaA, coxA, fbpA and gatB) of Wolbachia, we determined the supergroups in the specimens from bearded pigs and those of O. d. dewittei.
RESULTS: Onchocerca borneensis Uni, Mat Udin & Takaoka n. sp. is described on the basis of morphological characteristics and its genetic divergence from congeners. Molecular characteristics of the new species revealed its close evolutionary relationship with O. d. dewittei. Calculated p-distance for the cox1 gene sequences between O. borneensis n. sp. and O. d. dewittei was 5.9%, while that between O. d. dewittei and O. d. japonica was 7.6%. No intraspecific genetic variation was found for the new species. Wolbachia strains identified in the new species and O. d. dewittei belonged to supergroup C and are closely related.
CONCLUSIONS: Our molecular analyses of filarioids from Asian suids indicate that the new species is sister to O. d. dewittei. On the basis of its morphological and molecular characteristics, we propose to elevate O. d. japonica to species level as O. japonica Uni, Bain & Takaoka, 2001. Coevolutionary relationships exist between the Wolbachia strains and their filarial hosts in Borneo and Peninsular Malaysia.
METHODS: Genomic DNA of Indonesian black fly samples were extracted and sequenced, producing 86 COI sequences in total. Two hundred four COI sequences, including 118 GenBank sequences, were analysed. Maximum likelihood (ML) and Bayesian inference (BI) trees were constructed and species delimitation analyses, including ASAP, GMYC and single PTP, were performed to determine whether the species of Indonesian black flies could be delineated. Intra- and interspecific genetic distances were also calculated and the efficacy of COI sequences for species identification was tested.
RESULTS: The DNA barcodes successfully distinguished most morphologically distinct species (> 80% of sampled taxa). Nonetheless, high maximum intraspecific distances (3.32-13.94%) in 11 species suggested cryptic diversity. Notably, populations of the common taxa Simulium (Gomphostilbia) cheongi, S. (Gomphostilbia) sheilae, S. (Nevermannia) feuerborni and S. (Simulium) tani in the islands of Indonesia were genetically distinct from those on the Southeast Asian mainland (Malaysia and Thailand). Integrated morphological, cytogenetic and nuclear DNA studies are warranted to clarify the taxonomic status of these more complex taxa.
CONCLUSIONS: The findings showed that COI barcoding is a promising taxonomic tool for Indonesian black flies. The DNA barcodes will aid in correct identification and genetic study of Indonesian black flies, which will be helpful in the control and management of potential vector species.
METHODS: Blackfly larvae and pupae were sampled monthly from 58 sites between May 2011 and April 2013. Diversity parameters, seasonal abundance, regional distribution and frequency of species occurrence in stream sites were analyzed.
RESULTS: A total of 19,456 mature larvae representing 57 species, and belonging to six subgenera in the genus Simulium Latreille (s.l.), were found. The five predominant taxa were S. fenestratum (8.6%), the S. asakoae complex (8.3%), S. nakhonense (7.5%), the S. siamense complex (7.4%) and the S. doipuiense complex (6.7%). The most frequent taxa at all sites were the S. asakoae complex (84.5%), followed by S. fenestratum (82.8%), the S. siamense complex (75.9%), S. decuplum (60.3%), S. nakhonense (58.6%) and the S. tani complex (48.3%). The richness of regional species was highest (40 species) in the north and predominated in the cold season. However, blackflies in the south predominated during the hot season. The highest numbers of blackflies collected from central, northeastern, eastern and western regions of the country were observed in the rainy season. Overall, the mean number of blackflies collected across the six regions during the rainy and cold season had no statistically significant difference, but it differed significantly in the hot season.
CONCLUSIONS: Blackflies in Thailand were surveyed in all three seasons across six geographical regions. These findings demonstrated that blackfly communities at each stream site varied with seasonality, and the regional relative abundance of blackflies differed markedly in the hot season. It was also found that the occurrence and distribution of blackflies in each region were associated strongly with elevation.