Use of an Anopheles Salivary Biomarker to Assess Malaria Transmission Risk Along the Thailand-Myanmar Border.

Background: The modalities of malaria transmission along the Thailand-Myanmar border are poorly understood. Here we address the relevance of using a specific Anopheles salivary biomarker to measure the risk among humans of exposure to Anopheles bites. Methods: Serologic surveys were conducted from May 2013 to December 2014 in 4 sentinel villages. More than 9400 blood specimens were collected in filter papers from all inhabitants at baseline and then every 3 months thereafter, for up to 18 months, for analysis by enzyme-linked immunosorbent assay. The relationship between the intensity of the human antibody response and entomological indicators of transmission (human biting rates and entomological inoculation rates [EIRs]) was studied using a multivariate 3-level mixed model analysis. Heat maps for human immunoglobulin G (IgG) responses for each village and survey time point were created using QGIS 2.4. Results: The levels of IgG response among participants varied significantly according to village, season, and age (P<.001) and were positively associated with the abundance of total Anopheles species and primary malaria vectors and the EIR (P<.001). Spatial clusters of high-IgG responders were identified across space and time within study villages. Conclusions: The gSG6-P1 biomarker has great potential to address the risk of transmission along the Thailand-Myanmar border and represents a promising tool to guide malaria interventions.

Behavioral Action of Deltamethrin and Cypermethrin in Pyrethroid-Resistant Aedes aegypti (Diptera: Culicidae): Implications for Control Strategies in Thailand.

Contact irritant (locomotor excitation) and noncontact spatial repellent avoidance behavior to deltamethrin and cypermethrin at dosages 0.025, 0.05, and 0.1 g/m2 impregnated on papers were evaluated in the laboratory against deltamethrin- and cypermethrin-resistant field populations of female Aedes aegypti (L.) from Rayong and Chanthaburi Provinces, Thailand. Pyrethroid-resistant populations were compared with a susceptible laboratory strain (NIH-Thai) using an 'excito-repellency' (ER) test system. Both NIH-Thai and field mosquitoes had stronger contact irritancy responses compared to the relatively weak noncontact repellency effects. Contact assays with deltamethrin and cypermethrin at 0.1 g/m2 showed high escape rates for Rayong (80.1 and 83.4%, respectively) and Chanthaburi (84.6 and 73.1%, respectively) mosquitoes. Cypermethrin produced significantly different (P < 0.05) percent escape responses in contact tests between NIH-Thai and field mosquitoes. Only deltamethrin contact at 0.05 g/m2 produced a significant escape response (P < 0.001) between NIH-Thai and Rayong mosquitoes. These results suggest that there may not be an overall significant effect of background pyrethroid resistance on escape response, and the differences by comparisons may reflect inherent individual variation when using the ER bioassay system. The results show that pyrethroid resistance in Ae. aegypti does not appear to influence or reduce contact avoidance responses with the compounds tested. In particular, deltamethrin at 0.1 g/m2 was an effective contact irritant and toxic compound against pyrethroid-resistant populations of Ae. aegypti. Therefore, 0.1 g/m2 deltamethrin could be considered for residual applications of either fixed surfaces or materials (e.g., curtains) as a supplemental control measure against adult dengue vectors.

Anopheles Salivary Biomarker as a Proxy for Estimating Plasmodium falciparum Malaria Exposure on the Thailand-Myanmar Border.

Timely identification and treatment of malaria transmission "hot spots" is essential to achieve malaria elimination. Here we investigate the relevance of using an Anopheles salivary biomarker to estimate Plasmodium falciparum malaria exposure risk along the Thailand-Myanmar border to guide malaria control. Between May 2013 and December 2014, > 9,000 blood samples collected in a cluster randomized control trial were screened with serological assays to measure the antibody responses to Anopheles salivary antigen (gSG6-P1) and P. falciparum malaria antigens (circumsporozoite protein, merozoite surface protein 119 [MSP-119]). Plasmodium falciparum infections were monitored through passive and active case detection. Seroprevalence to gSG6-P1, MSP-119, and CSP were 71.8% (95% Confidence interval [CI]: 70.9, 72.7), 68.6% (95% CI: 67.7, 69.5), and 8.6% (95% CI: 8.0, 9.2), respectively. Multivariate analysis showed that individuals with the highest Ab response to gSG6-P1 had six times the odds of being positive to CSP antigens (P < 0.001) and two times the odds of P. falciparum infection compared with low gSG6-P1 responders (P = 0.004). Spatial scan statistics revealed the presence of clusters of gSG6-P1 that partially overlapped P. falciparum infections. The gSG6-P1 salivary biomarker represents a good proxy for estimating P. falciparum malaria risk and could serve to implement hot spot-targeted vector control interventions to achieve malaria elimination.

Climate associated size and shape changes in Aedes aegypti (Diptera: Culicidae) populations from Thailand.

In spite of the adult body size variability of Aedes aegypti (Linnaeus) and its likely association with life history and vectorial capacity, the causes of size variation itself have been only partially identified. In particular, possible important factors such as climatic variation have not received much attention. The objective of this 2-year study was to describe from field collections the relationship of Ae. aegypti metric properties with available climatic data. The study took place in a dengue hyperendemic area of Thailand. Fourth instar larvae (L(4)) and pupae were collected from the same breeding places allowing the comparisons between seven successive collections, four in 2007 and three in 2008. Climatic data were relative humidity (RH) and temperature (T). They were considered for the periods covering either the pre-imaginal development or, assuming heritability of size, the previous generation. The pre-imaginal period was further subdivided into embryonic and larval phases of development. Size was estimated by traditional and geometric techniques, the latter based on 18 landmarks collected at the intersections of veins also allowing estimation of shape. The shape variation of the wing followed similar patterns as for size and was shown to be a passive allometric change. No significant correlation of size or shape could be disclosed with T. In contrast, significant correlation with RH was found during two periods of examination: (i) the period affecting the generation previous to the time of collection, suggesting possible selective mechanisms on genitors, and (ii) the one occurring during pre-imaginal development. The subdivision of the latter into embryonic and larval phases allowed to evidence a possible selecting effect on embryonic development. The selection would act through the resistance to water loss which is known to depend on the relative surface of the cuticle. In conclusion, our data highlight the importance of the emerged period of Ae. aegypti eggs as a critical time for the size of future adults, and point to the relative humidity as the likely selecting factor.

Insecticide susceptibility and selection for resistance in a population of Aedes aegypti from Ratchaburi province, Thailand.

The insecticide susceptibility of Aedes aegypti larvae and adults from four areas of Ratchaburi Province, Thailand was investigated using World Health Organization standard procedures. The larvae of Ae. aegypti in all areas were found to be susceptible to temephos. The strain with the highest level of temephos resistance was subjected to temephos selection. The resistance ratios increased to 2.74-fold from their parental non selected strain after 5 generations of selection. For the adult of Ae. aegypti, the study was conducted to determine the effects of 0.75 % permethrin and 0.05 % deltamethrin impregnated paper selection on the strain having the highest with the highest LT50 level. The results showed the increase in the LT50 to 7.46 and 1.18-fold after the third selected generation with 0.75 % permethrin and 0.05 % deltamethrin respectively. Mortality rates after exposing adults to discriminating concentrations showed that field populations in Ratchaburi Province were resistant to permethrin and deltamethrin. Alternative non-chemical measures need to be used in new of the emergence of resistance in the mosquito population.

Biochemical detection of pyrethroid resistance mechanism in Aedes aegypti in Ratchaburi province, Thailand.

The emergence of insecticide resistance in mosquito vectors was an important issue to be considered as one of factors influencing the success of vector control. The early detection of resistance could help the health personnel to plan and select appropriate alternative control measures or insecticide for effective control. Therefore biochemical assay of enzymes in mosquito was conducted to detect the emergence of insecticide resistance and to define the machanisms involved in pyrethroid resistance. Adults of Aedes aegypti from two localtities in Ratchaburi province were subjected to permethrin and deltamethrin selection in laboratory. After three generations of selection, permethrin-selected and deltamethrin-selected strains were established. Their LT 50 increased to 7.46 and 1.18 folds in the F3 strains that were selected with permethrin and deltamethrin respectively. The enzymes of these mosquitoes were assayed biochemically to study the mechanisms of resistance. The results revealed significant increase of esterase activity and monooxygenase levels in both strains when compared with labolatory susceptible strain. Glutathione-S-transferase activity was found to increase in permethrin-selected strain but not in deltamethrin-selected strain. This suggested that not only esterase and monooxygenase but also glutathione-S-transferase were associated with permethrin resistance in Ae. aegypti. The exposing of permethrin-selected and deltamethrin-selected mosquitoes to diagnostic concentration of permethrin (0.75%) and deltamethrin (0.05%) indicated no cross resistance for permethrin to deltamethrin while slight cross resistance from deltamethrin to permethrin was evident. It seemed that glutathione S-tranferase was not associated with cross resistance since its activity in deltamethrin-selected strain remained unchanged as compared with that of laboratory susceptible strain.