Sungsit Sungvornyothin

Trophic behavior and biting activity of the two sibling species of the Anopheles minimus complex in western Thailand

ABSTRACT The trophic behavior and host preference of two sibling species, Anopheles minimus s.s. (= An. minimus species A) and species C, were observed during a two-year period at Pu Teuy Village, Sai Yok District, Kanchanaburi Province, western Thailand. Anopheles minimus s.s. and species C were more prevalent during the hot and wet periods of the year. Both species demonstrated exophagic and zoophilic activities. Feeding activity of An. minimus C was unique compared to An. minimus sensu lato from other localities in Thailand. Outdoor blood feeding by An. minimus C occurred throughout the night with one distinct feeding peak immediately after sunset (1800 h), whereas indoor feeding showed two small peaks at 2000 and 2400 h. The small number of An. minimus s.s collected during this study precluded a determination of peak activity patterns. A better understanding of mosquito behavior related to host and patterns of feeding activity will facilitate and improve the efficiency of vector control operations.

Biting Activity and Host Preference of the Malaria Vectors Anopheles maculatus and Anopheles sawadwongporni (Diptera: Culicidae) in Thailand

ABSTRACT: Adult mosquitoes in the Anopheles maculatus group were surveyed from different regions of Thailand and five different species were morphologically identified, including Anopheles maculatus, Anopheles sawadwongporni, Anopheles notanandai, Anopheles dravidicus, and Anopheles willmori. Blood-feeding activity and host preference of two species, Anopheles maculatus and Anopheles sawadwongporni, were observed during a one-year period at Pu Teuy Village, Sai Yok District, Kanchanaburi Province, west-central Thailand. Both species were more prevalent during the wetter period of the year and each had a greater predilection to feed on cattle than humans. Primary feeding activity occurred between 20:00–23:00 and a smaller peak at 01:00–03:00. Findings are discussed relative to the importance of these two vectors for malaria transmission in Pu Teuy.

Seasonal Abundance and Bloodfeeding Activity of Anopheles dirus Sensu Lato in Western Thailand

Abstract Bloodfeeding activity, host preference, and seasonal abundance of the Anopheles dirus complex in relation to environmental factors were measured during a 2-yr period at Pu Teuy Village, Sai Yok District, Kanchanaburi Province, western Thailand. Anopheles dirus s.l. was found more abundant during the wet season compared with the dry and hot seasons. Bloodfeeding by An. dirus s.l. commenced immediately after sunset with a distinct peak of activity at 2000 h. Anopheles dirus s.l. fed more readily on cattle than on humans, demonstrating a significant but slightly stronger zoophilic behavior (P  =  0.02). The biting activity of An. dirus s.l. in Pu Teuy appears to differ from other localities in Thailand, suggesting the existence of significant biological variability within the species complex depending on the local circumstances.

HOW RELIABLE IS THE HUMERAL PALE SPOT FOR IDENTIFICATION OF CRYPTIC SPECIES OF THE MINIMUS COMPLEX

ABSTRACT The Anopheles minimus Complex Theobald (Diptera: Culicidae) is composed of the 3 sibling species A, C, and E. The malaria vectors An. minimus A and C are distributed over the Southeast Asian region, whereas species E is restricted to the Ryukyu Japanese islands. Because species A and C can be sympatric and present specific behaviors and have a role in malaria transmission, it is important to differentiate them. The literature mentioned the presence of a presector pale spot on the wing costa of An. minimus A, whereas species C may exhibit both presector and humeral pale spots. However, the reliability of their diagnostic power has not been established over large temporal and geographic surveys. From the analyses of 9 populations throughout Southeast Asia, including published data and field populations from 2 sites in Thailand, we showed that the wing patterns present spatial and temporal variations that make these two morphological characters unreliable for the precise identification of An. minimus A and C. Therefore, molecular identification remains the most efficient method to obtain an unambiguous differentiation of these 2 species. Correct species identification is essential and mandatory for any relevant study on the Minimus Complex and for the application of successful control strategies.

An Improved Experimental Hut Design for the Study of Aedes aegypti (Diptera: Culicidae) Movement Patterns in Thailand

1Department of Entomology, Kasetsart University, Bangkok 10900, Thailand 2Department of Preventive Medicine and Biometrics, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, U.S.A. 3Office of Disease Prevention and Control No 10, 18 Boonrungrit Rd., Chiangmai 50200, Thailand 4Division of Biostatistics and Computer, Faculty of Liberal Arts and Science, Kasetsart University, Kamphaengsean Campus, Nakhon Pathom 73140, Thailand 5Department of Medical Entomology, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand

Comparison of Aedes aegypti (Diptera: Culicidae) Resting Behavior on Two Fabric Types Under Consideration for Insecticide Treatment in a Push-Pull Strategy

ABSTRACT Aedes aegypti (L.), the primary vector of dengue and dengue hemorrhagic fever, breeds and rests predominately inside human dwellings. With no current vaccine available, vector control remains the mainstay for dengue management and novel approaches continue to be needed to reduce virus transmission. This requires a full understanding of Ae. aegypti ecology to design effective strategies. One novel approach is the use of contact irritants at target resting sites inside homes to make the surface unacceptable and cause vectors to escape before biting. The objective of the current study was to observe indoor resting behavior patterns of female Ae. aegypti within experimental huts in response to two fabrics under consideration for insecticide treatment: cotton and polyester. Results indicate that fabric type, coverage ratio of dark to light fabric and placement configuration (vertical vs. horizontal) all influenced the resting pattern of mosquito cohorts. Findings from this study will guide evaluations of a push-pull strategy designed to exploit contact irritant behaviors and drive Ae. aegypti out of homes prefeeding.

A single-round multiplex PCR assay for the identification of Anopheles minimus related species infected with Plasmodium falciparum and Plasmodium vivax

This study aimed to develop a single-round multiplex PCR method for the identification of Anopheles minimus complex (An. minimus and Anopheles harrisoni) and Anopheles aconitus subgroup (An. aconitus and Anopheles varuna), and for the simultaneous detection of Plasmodium falciparum and Plasmodium vivax in these vectors. Five primers were created for a single-round multiplex PCR assay to identify four anopheline mosquitoes combined with three Plasmodium primers for the detection of P. falciparum and P. vivax in vectors. The four species of anopheline vectors and two Plasmodium species, P. falciparum and P. vivax, could be identified by the combination of eight primers in the single-round multiplex PCR assay. The amplified species-specific products were 380bp for An. minimus, 180bp for An. harrisoni, 150bp for An. aconitus, 310bp for An. varuna, 276bp for P. falciparum, and 300bp for P. vivax. The sensitivities were 0.5pg/μl (25sporozoites/μl) for P. falciparum DNA and between 0.5 and 5pg/μl (25-250sporozoites/μl) for P. vivax DNA. Furthermore, this developed method could be used to identify field caught An. minimus complex, An. aconitus subgroup from Thailand and Lao PDR. Also, it was successfully used to identify the species An. minimus, An. harrisoni, An. aconitus and An. varuna and to detect and identify P. falciparum and P. vivax in caught anopheline mosquitoes. The sensitivity of this method was high for simultaneous detection of P. falciparum and P. vivax in anopheline mosquitoes.

Functional expression and molecular characterization of Culex quinquefasciatus salivary α-glucosidase (MalI)

Salivary α-glucosidases (MalI) have been much less characterized when compared with midgut α-glucosidases, which have been studied in depth. Few studies have been reported on the partial characterization of MalI, but no clear function has been ascribed. The aim of this study is to purify and characterize the recombinant Culex quinquefasciatus (CQ) α-glucosidase expressed in Pichia pastoris. The cDNA encoding mature Cx. quinquefasciatus α-glucosidase gene with polyhistidine tag (rCQMalIHis) was successfully cloned into the expression vector, pPICZαB, designated as pPICZαB/CQMalIHis. The activity of recombinant rCQMalIHis expressed in P. pastoris could be detected at 3.75U/ml, under optimal culture conditions. The purified rCQMalIHis showed a single band of molecular weight of approximately 92kDa on SDS-PAGE. After Endoglycosidase H digestion, a single band at 69kDa was found on SDS-PAGE analysis, suggesting that rCQMalIHis is a glycoprotein. Additionally, tryptic digestion and LC-MALDI MS/MS analysis suggested that the 69kDa band corresponds to the Cx. quinquefasciatus α-glucosidase. Thus, rCQMalIHis is a glycoprotein. The rCQMalIHis exhibited optimum pH and temperature at 5.5 and 35°C, respectively. The catalytic efficiency (kcat/Km) of the purified rCQMalIHis for maltotriose is higher than those for sucrose, maltotetraose, maltose and p-nitrophenyl-α-glucoside, indicating that the enzyme prefers maltotriose. Additionally, the rCQMalIHis is significantly inhibited by d-gluconic acid δ-lactone, but not by Mg(2+), Ca(2+) and EDTA. The rCQMalIHis is strongly inhibited by acarbose with IC50 67.8±5.6nM, but weakly inhibited by glucose with IC50 115.9±7.3mM.

Diversity Of Mosquito Species Ovipositing In Different Zones of Light Intensity Within Limestone Caves In Thailand

Climate change and human activity affect the geographical and annual distribution and population abundance of mosquitoes. As natural habitats are reduced, it is hypothesized that mosquitoes may seek refuge in more stable environments such as cave habitats. Therefore, we explored the species diversity of mosquitoes exploiting cave habitats in Thailand. Ten species belonging to 4 genera were collected, of which none were considered to be true cave-dwelling species (Troglobiont). The known cavernicolous species, Aedes cavaticus, was observed to oviposit outside of the cave and therefore should be categorized as a subtroglophilic species. Other species were also oviposited inside the cave but should be regarded as trogloxenic species. There was no clear association between environmental factors and mosquito abundance inside the 4 limestone caves, except for Ae. cavaticus, which was positively correlated with rainfall. This study indicates that different biotic or abiotic factors may be involved in mosquito oviposition site selection inside caves.