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For over 50 years, biological control of mosquito larvae has depended mainly on plant extracts, fish, bacteria, protozoa, filamentous fungi, viruses or nematodes. In this study, we screened 143 mushroom samples from 44 confirmed species in Thailand for their mosquito larvicidal activity. One g% (w/v) aqueous extracts of dried powdered mushroom samples were tested against 3rd stage Aedes aegypti larvae. Four mushroom species, namely, Thaeogyroporus porentosus, Xylaria nigripes, Chlorophyllum sp and Steccherinum sp, and two unidentified species showed larvicidal mortality ranging from 10%-70% and 18%-90% for 24- and 48-hour exposure time, respectively. Steccherinum sp aqueous crude extract, after 48-hour exposure, did not show any larvicidal activity at 1,000 ppm, whereas that from ethanol, after 24-hour exposure, had 50% and 90% lethal concentration of 203 ppm and 412 ppm, respectively, with higher levels of mortality after 48- hour exposure. This is the first report of mosquito larvicidal properties of Thai mushroom extracts.

Variation in vectorial capacity for human malaria among Anopheles mosquito species is determined by many factors, including behavior, immunity, and life history. To investigate the genomic basis of vectorial capacity and explore new avenues for vector control, we sequenced the genomes of 16 anopheline mosquito species from diverse locations spanning ~100 million years of evolution. Comparative analyses show faster rates of gene gain and loss, elevated gene shuffling on the X chromosome, and more intron losses, relative to Drosophila. Some determinants of vectorial capacity, such as chemosensory genes, do not show elevated turnover but instead diversify through protein-sequence changes. This dynamism of anopheline genes and genomes may contribute to their flexible capacity to take advantage of new ecological niches, including adapting to humans as primary hosts., (Copyright © 2015, American Association for the Advancement of Science.)

Objectives: To develop rapid monitoring tools to detect the F1534C permethrin-resistance mutation in domain IIIS6 of the Aedes aegypti voltage-gated sodium channel gene and determine the frequency and distribution of this mutation in Thailand., Methods: A TaqMan SNP genotyping and an allele specific PCR (AS-PCR) assay were developed and validated by comparison with DNA sequencing of homozygous susceptible and homozygous resistant laboratory strains, their reciprocal-cross progenies, and field-caught mosquitoes. To determine the resistance phenotype of wild-caught A. aegypti, mosquitoes were exposed to 0.75% permethrin paper. The AS-PCR assay was used to screen 619 individuals from 20 localities throughout Thailand., Results: Overall, both assays gave results consistent with DNA sequencing for laboratory strains of known genotype and for wild-caught A. aegypti. The only slight discrepancy was for the AS-PCR method, which overestimated the mutant allele frequency by 1.8% in wild-caught samples. AS-PCR assays of permethrin-exposed samples show that the mutant C1534 allele is very closely associated with the resistant phenotype. However, 19 permethrin-resistant individuals were homozygous for the wild-type F1534 allele. DNA sequencing revealed all these individuals were homozygous for two other mutations in domain II, V1016G and S989P, which are known to confer resistance (Srisawat et al. 2010). The F1534C mutation is widespread in Thailand with mutant allele frequencies varying among populations from 0.20 to 1.00., Conclusions: These assays can be used for the rapid detection of the F1534C resistance mutation in A. aegypti populations. The F1534C, and other, mutations underlie an extremely high prevalence of pyrethroid resistance in Thailand., (© 2011 Blackwell Publishing Ltd.)

Nine colonies of five sibling species members of Anopheles barbirostris complexes were experimentally infected with Plasmodium falciparum and Plasmodium vivax. They were then dissected eight and 14 days after feeding for oocyst and sporozoite rates, respectively, and compared with Anopheles cracens. The results revealed that Anopheles campestris-like Forms E (Chiang Mai) and F (Udon Thani) as well as An. barbirostris species A3 and A4 were non-potential vectors for P. falciparum because 0% oocyst rates were obtained, in comparison to the 86.67-100% oocyst rates recovered from An. cracens. Likewise, An. campestris-like Forms E (Sa Kaeo) and F (Ayuttaya), as well as An. barbirostris species A4, were non-potential vectors for P. vivax because 0% sporozoite rates were obtained, in comparison to the 85.71-92.31% sporozoite rates recovered from An. cracens. An. barbirostris species A1, A2 and A3 were low potential vectors for P. vivax because 9.09%, 6.67% and 11.76% sporozoite rates were obtained, respectively, in comparison to the 85.71-92.31% sporozoite rates recovered from An. cracens. An. campestris-like Forms B and E (Chiang Mai) were high-potential vectors for P. vivax because 66.67% and 64.29% sporozoite rates were obtained, respectively, in comparison to 90% sporozoite rates recovered from An. cracens.

The genetic structure of four populations of the malarial vector Anopheles scanloni in Thailand was studied using mitochondrial DNA sequences. Four highly divergent lineages were observed, all with signals of population expansion. Since An. scanloni is restricted to 'islands' of limestone karst habitat, this suggests there is a metapopulation-type dynamic in this species, with restricted gene flow, extinctions and drift all contributing to lineage divergence. Historical environmental change and marine transgressions may also have contributed to population extinction, expansion and divergence. Although there is some current gene flow inferred between nearby populations, it is extremely restricted between the northern and southern populations, which also differed by one fixed polymorphism at the ITS2 rDNA locus. Crossing experiments showed no post-mating barriers existing between the north and the south, but the lack of gene flow between these populations could ultimately result in speciation and has implications for malaria control strategies.

The Anopheles annularis group of subgenus Cellia Theobald (Diptera: Culicidae) includes five currently recognized species in southern Asia: An. annularis Van der Wulp, Anopheles nivipes (Theobald) and Anopheles philippinensis Ludlow, which are widespread in the region, Anopheles pallidus Theobald, which is known in Sri Lanka, India and Myanmar, and Anopheles schueffneri Stanton, which occurs in Java and Sumatra. Identification of the four mainland species based on morphology is problematic. In view of the fact that the three widespread species are variously involved in malaria transmission in different parts of the region, we developed a species-specific polymerase chain reaction assay based on rDNA internal transcribed spacer 2 (ITS2) sequences to facilitate entomological and epidemiological studies of the four species. The method proved to be reliable when tested over a wide geographical area.

The objective of this study was to test for associations between land cover data and the presence of mosquito larvae of the genera Aedes Meigen and Anopheles Meigen in northern Thailand at the landscape scale. These associations were compared with associations between larval habitat variables and the presence of mosquito larvae at a finer spatial scale. Collection data for the larvae of one Aedes species and three species-groups of Anopheles, all of which are involved in pathogen transmission, were used. A variety of northern Thai landscapes were included, such as upland villages, lowland villages and peri-urban areas. Logistic regression was used to evaluate associations. Generally, land cover and landscape variables explained the presence of larvae as well as did larval habitat variables. Results were best for species/species-groups with specific habitat requirements. Land cover variables act as proxies for the types of habitat available and their attributes. Good knowledge of the habitat requirements of the immature stages of mosquitoes is necessary for interpreting the effects of land cover.

Four laboratory-raised colonies of two karyotypic forms of Anopheles aconitus, i.e., Form B (Chiang Mai and Phet Buri strains) and C (Chiang Mai and Mae Hong Son strains), were experimentally infected with Plasmodium falciparum and P. vivax using an artificial membrane feeding technique and dissected eight and 12 days after feeding for oocyst and sporozoite rates, respectively. The results revealed that An. aconitus Form B and C were susceptible to P. falciparum and P. vivax, i.e., Form B (Chiang Mai and Phet Buri strains/P. falciparum and P. vivax) and Form C (Chiang Mai and Mae Hong Son strains/P. vivax). Comparative statistical analyses of the oocyst rates, average number of oocysts per infected midgut and sporozoite rates among all strains of An. aconitus Form B and C to the ingroup control vectors, An. minimus A and C, exhibited mostly no significant differences, confirming the high potential vector of the two Plasmodium species. The sporozoite-like crystals found in the median lobe of the salivary glands, which could be a misleading factor in the identification of true sporozoites in salivary glands were found in both An. aconitus Form B and C.

At least three members (species A, C, and E) of the Anopheles minimus complex have been described in the Orient. This study investigated the specific status of An. minimus collected in the southern part of Taiwan by crossing experiments with species A from Thailand and species E from Japan. Crosses between Taiwan An. minimus and species A revealed genetic compatibilities. Post-zygotic isolation was observed in crosses between Taiwan An. minimus and species E. Hybrid progeny were only obtained from Taiwan female X species E male. F2 hybrid progeny were not obtained, since the hybrid males were sterile or almost sterile, with atrophied testes or abnormal spermatozoa. The hybrid females backcrossed with either Taiwan F1 progeny and species E males, and laid eggs with lower fertility and viability. This study supports previous published data regarding the analysis of the D3 region of the 28S gene of ribosomal DNA that An. minimus species A is indigenous to Taiwan. Whether other members of the An. minimus complex exist in Taiwan is not conclusive and needs more study.

Midgut proteolytic enzymes contribute to the success or failure of Plasmodium infection of the mosquito. The present study investigated trypsin and aminopeptidase activities in the midgut of two strains of Anopheles dirus selected for susceptibility and refractoriness to Plasmodium yoelii nigeriensis. At intervals of 6 hours following a bloodmeal, the midguts of fully engorged female mosquitos were dissected, homogenized, and assayed for enzyme activity. No differences trypsin activity (nmole/min) were observed between the two strains throughout the course of blood digestion. By contrast, the aminopeptidase activity measured at 0 to 18 hours post-feeding was the same for the two strains, but at 24, 30 and 36 hours significantly less activity was observed in the refractory females. The results suggest neither trypsin nor aminopeptidase plays a role in the limitation of parasite development.

We surveyed Nongceng, a village in a south-eastern province of Lao PDR, for malaria and its vectors. Nongceng is situated in a basin and surrounded by rice fields. In February 1998 (dry season), 28.6% of 126 villagers were infected with malaria, and in September 1998 (rainy season), 16.3% of 147 villagers. The prevalence of malaria infection was consistently high in children under 10, and the predominant malaria species was Plasmodium falciparum. In brief surveys of the mosquitoes performed on the same day as the malaria surveys, 2007 Anopheles females from 12 species were collected by means of human bait, animal bait and resting collections. Of the vector species known to be important in transmitting malaria in neighbouring Thailand - An. minimus, An. dirus, and An. maculatus groups - only An. minimus was found. Its density was, however, very low in both seasons and it was therefore unlikely to be the vector. In fact, An. nivipes accounted for more than 65% of all mosquitoes collected and was the most common species collected from human baits. The results of this study show that endemic areas of malaria in Lao PDR are not necessarily related to forest. Rather, An. nivipes is suspected to be the most important vector.

Wild caught zoophilic Anopheles and suspected malaria vector species collected in northwest Thailand were experimentally infected with local human malaria parasites using a membrane feeding. One week post-feeding a number of mosquitos were dissected for oocyst examination. The remainder were kept for another one week or more, and then the salivary glands were examined for the presence of sporozoites. The results revealed that An. vagus, An. kochi and An. annularis were susceptible to both Plasmodium falciparum and P. vivax whereas An. barbirostris and An. sinensis were susceptible to only P. vivax. The non-susceptibility to P. falciparum of these two mosquito species may indicate their poor vector status of this malaria species in the field.

From February 1987 to January 1988, biological and ecological studies were made to obtain the basic knowledge of Japanese encephalitis vectors, Culex tritaeniorhynchus, Cx. gelidus, and Cx. fuscocephala, in Amphoe Muang, Chiang Mai, northern Thailand. The following results were found. Peaks in the population densities of the vectors as measured four times a month, by UV-light trap and human-baited trap collections, occurred during rainy season. The JE vectors in rural areas showed a sharp rise in the population in July when most of the rice fields were ploughed and a marked decline in mosquito population densities occurred after transplanting in August when the fields were flooded. The average number of larvae plus pupae per m2 in rice fields was highest in July when the fields were ploughed, but in the period from transplanting to harvesting (August to November), the densities were very low. Daily survival rates of the adult females, as estimated from parous rates, were mostly as high as 0.7 throughout the year.

Variation in vectorial capacity for human malaria among Anopheles mosquito species is determined by many factors, including behavior, immunity, and life history. To investigate the genomic basis of vectorial capacity and explore new avenues for vector control, we sequenced the genomes of 16 anopheline mosquito species from diverse locations spanning ~100 million years of evolution. Comparative analyses show faster rates of gene gain and loss, elevated gene shuffling on the X chromosome, and more intron losses, relative to Drosophila. Some determinants of vectorial capacity, such as chemosensory genes, do not show elevated turnover but instead diversify through protein-sequence changes. This dynamism of anopheline genes and genomes may contribute to their flexible capacity to take advantage of new ecological niches, including adapting to humans as primary hosts.