Frequency of pyrethroid resistance in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in Thailand.

Thirty-two Aedes aegypti populations collected throughout Thailand and five populations of Aedes albopictus from southern Thailand were subjected to standard WHO contact bioassays to assess susceptibility to three commonly used synthetic pyrethroids: permethrin, deltamethrin, and lambda-cyhalothrin. A wide degree of physiological response to permethrin was detected in Ae. aegypti, ranging from 56.5% survival (Lampang, northern Thailand) to only 4% (Kalasin in northeastern and Phuket in southern Thailand). All 32 populations of Ae. aegypti were found to have evidence of incipient resistance (62.5%) or levels of survival deemed resistant (37.5%) to permethrin. Four populations of Ae. albopictus were found with incipient resistance (97 - 80% mortality) and one with resistance (< 80%) to permethrin. The majority of Ae. aegypti populations (68.7%) was susceptible (> 98% mortality) to deltamethrin, with incipient resistance (observed 97-82% mortality) in other localities. In contrast, all populations of Ae. aegypti were completely susceptible (100% mortality) to the recommended operational dosage of lambda-cyhalothrin. All five populations of Ae. albopictus were found completely susceptible to both deltamethrin and lambda-cyhalothrin. Evidence of defined incipient or resistance to synthetic pyrethroids mandates appropriate response and countermeasures to mitigate further development and spread of resistance. In light of these findings, we conclude that routine and comprehensive susceptibility monitoring of dengue mosquito vectors to synthetic pyrethroids should be a required component of resistance management policies and disease control activities.

Geographic distribution of stomoxyine flies (Diptera: Muscidae) and diurnal activity of Stomoxys calcitrans in Thailand.

Stomoxyine flies (Stomoxys spp.) were collected in 10 localities of Thailand using the Vavoua traps. These localities represented four major ecological settings, as follows: small local dairy farms, large industrial dairy farms, a national park, and one elephant conservation area. Three species of stable flies were identified in the following proportions: Stomoxys calcitrans (91.5%), Stomoxys indicus (7.9%), and Stomoxys sitiens (0.6%). The number of flies collected differed significantly among collection sites (chi2 = 360.15, df=3, P < 0.05). The greatest number of stomoxyine flies was captured in dairy farms. Seasonal and daily activity of S. calcitrans was observed during a 1-yr period at two selected locations (Dairy Farming Promotion Organization of Thailand and Khao Kheow Open Zoo). S. calcitrans was more abundant during the rainy season (March-September), but was not associated with the total rainfall (r2 = 0.0002, P > 0.05). Peak of daily flight activity of males S. calcitrans was at 1000 and 1600 h, whereas females showed an increase of activity all along the day until 1600 h. A better understanding of stomoxyine fly behavior related to patterns of daily activity will facilitate and improve the efficiency of fly control measures in private and government sectors.

Human-landing patterns of Anopheles dirus sensu lato (Diptera: Culicidae) in experimental huts treated with DDT or deltamethrin.

Anopheles dirus females landing on humans inside experimental huts treated with residual applications of DDT or deltamethrin were observed during the wet season in Pu Teuy Village, Kanchanaburi Province, western Thailand. Two identical experimental huts were constructed in the fashion of typical local rural Thai homes. Pretreatment (baseline) human-landing collections (HLC) in both huts showed an early evening peak of activity between 1900 and 2000 h with no significant difference in numbers of mosquitoes captured between huts over a period of 30 collection nights. During posttreatment HLC, female mosquitoes continued to show greater landing activity inside huts fitted with insecticide-treated panels during the first half of the evening compared with the second half. A greater number (proportion) of An. dirus females landed on humans in the hut treated with deltamethrin compared with DDT. Comparing pre- and posttreatment HLC, the DDT-treated hut showed a 79.4% decline in attempted blood feeding, whereas exposure to deltamethrin resulted in a 56.3% human-landing reduction. An odds ratio was performed to demonstrate the relative probability (risk) of mosquitoes entering and attempting to blood feed in the two treated huts compared with untreated control huts. Mosquitoes were approximately 0.47 times less likely to land on humans inside a DDT-treated hut compared with the deltamethrin-treated hut. Although both chemicals exerted strong excitatory responses, DDT appears to have a more pronounced and significant (P = 0.002) effect on behavior than deltamethrin, resulting in greater movement away from the insecticide source and thus potential reduction of blood-feeding activity.

Population structure of Stomoxys calcitrans (Diptera: Muscidae) from nine regions of Thailand.

Starch gel electrophoresis of isozymes was used to estimate gene flow among nine populations of Stomoxys calcitrans (L.) (Diptera: Muscidae) from Thailand. Of the 13 putative loci, nine polymorphic loci were detected. Limited genetic differentiation among populations was observed (F(ST) = 0.060). The highest level of polymorphism was observed in flies from eastern Trat and northern Chiang Mai provinces (69.2%), whereas the lowest level of polymorphism was seen in flies from central Saraburi Province (23.1%). Gene flow between populations varied from 3.27 to 27.53 reproductive migrants per generation. Among the nine populations sampled, no correlation was seen between genetic and geographical distances showing that sampled S. calcitrans fit closely in the same cluster taxa. The electrophoresis of ten isozymes shows a genetic homogeneity of S. calcitrans populations at the scale of Thailand.

Biting patterns of Anopheles minimus complex (Diptera: Culicidae) in experimental huts treated with DDT and deltamethrin.

Biting patterns of natural populations of Anopheles minimus s.l. females entering experimental huts treated with DDT and deltamethrin were carried out at Pu Teuy Village, Sai Yok District, Kanchanaburi Province, western Thailand. Two experimental huts, control and treatment, were constructed in the fashion of local Thai homes. Pre-spray biting activity of An. minimus females peaked at 19:00-22:00. Post-treatment exposure continued to show greater landing activity during the first half of the evening. An overall greater proportion of An. minimus females entered the hut treated with deltamethrin compared to DDT. The hut fitted with DDT-treated net panels showed a 71.5% decline in attempted blood feeding, whereas exposure to deltamethrin-treated panels resulted in a 42.8% human-landing reduction. DDT exhibited significantly more pronounced (P < 0.05) effects in overall reduction of biting activity than did deltamethrin.

Influence of nutritional and physiological status on behavioral responses of Aedes aegypti (Diptera: Culicidae) to deltamethrin and cypermethrin.

Excito-repellency responses of Aedes aegypti (L.) exposed to deltamethrin and cypermethrin were assessed using an excito-repellency test system. Contact irritancy and non-contact repellency assays compared non-bloodfed (unfed) parous (post-gravid), nulliparous, early blood-fed, late blood-fed, sugar-fed, and unmated female mosquitoes for behavioral responses based on nutritional and physiological conditions at the time of testing. Rates of escape during contact exposure with either compound were most pronounced in parous mosquitoes, followed by unmated mosquitoes, when compared to other conditional states. Significantly higher numbers of parous females also escaped from control chambers compared to other cohorts (P <0.05). Irritability of blood- and sugar-fed mosquitoes was noticeably suppressed. We conclude that nutritional and physiological conditions (including age) of mosquitoes at the time of testing can significantly influence behavioral responses (excito-repellency) to insecticides. The findings indicate that whether due to chronological age, nutrition, physiological state, or innate (circadian) activity patterns, careful consideration must be given to the selection of appropriate conditioned mosquitoes for testing.

Susceptibility and avoidance behavior by Culex quinquefasciatus Say to three classes of residual insecticides.

The behavioral responses ofthree colonized strains of Culex quinquefasciatus, two from recent field collections in Thailand (Nonthaburi and Mae Sot) and one from a long-established colony from the National Institute of Health (NIH), Ministry of Public Health, Thailand, were compared during and after exposure to deltamethrin (0.02 g/m2), propoxur (0.2 g/m2), and fenitrothion (0.2 g/m2) using an excito-repellency escape chamber system. We observed striking differences in behavioral response and excito-repellency between mosquito strains and test compounds. Greater escape responses were observed in the NIH strain during direct contact with deltamethrin and fenitrothion compared with the two field populations. Deltamethrin was the most irritant, followed by fenitrothion. Escape responses with propoxur were significantly delayed but increased slightly towards the end of the 30-min exposure period, more notably in the Nonthaburi strain (P < 0.05). Non-contact repellent responses were generally much weaker than irritancy, with the greatest escape response seen with NIH and Nonthaburi. Deltamethrin showed the weakest repellent response overall (< 10% escape), while propoxur again demonstrated a delayed effect (NIH and Mae Sot) before escape occurred. We conclude that irritant and repellent behavioral responses by Cx. quinquefasciatus are important components for assessing the impact of residual spraying in mosquito control programs. A better understanding of chemical properties that elicit behavioral responses in mosquitoes should be considered in formulating control strategies designed to control mosquitoes or mitigate disease transmission risk.

Behavioral responses of Aedes aegypti (Diptera: Culicidae) exposed to deltamethrin and possible implications for disease control.

Behavioral responses of nine Aedes aegypti (L.) strains, six from recent field collections and three from the long-established laboratory colonies, were tested under laboratory-controlled conditions by using an excito-repellency test system. All nine strains showed significant behavioral escape responses when exposed to deltamethrin at the standard field dose (0.02 g/m2), regardless of background insecticide susceptibility status (susceptible or tolerant/resistant). Insecticide contact irritancy played a predominate role in overall female mosquito escape responses, whereas noncontact repellency was not observed at levels significantly different from paired noncontact control tests (P > 0.01). Among the six field populations, the Jakarta (Indonesia) Toba (north Sumatra), and Bangkok female mosquitoes showed rapid exit (>78%) during 30 min of direct contact with insecticide-treated surfaces, whereas the other three strains demonstrated only moderate escape responses (32-56%) from the chambers. Moderate escape responses during direct insecticidal contact also were observed in the three laboratory test populations (44- 60%). Higher percentage of mortality was observed from laboratory strains (8-33%) that failed to escape compared with nonescape females of field strains (2-16%), possibly a reflection of background deltamethrin susceptibility status. We conclude that contact irritancy is a major behavioral response of Ae. aegypti when exposed directly to deltamethrin and that rapid flight escape from areas exposed to space sprays or surfaces treated with residual pyrethroids could have a significant impact on the effectiveness of adult mosquito control and disease transmission reduction measures.

Larval habitats and distribution patterns of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse), in Thailand.

This study was conducted to survey larval breeding habitats and to obtain larval abundance during the dry period covering all 5 geographical zones of Thailand. Our results indicated Aedes aegypri is prevalent all over the country, whereas Aedes albopictus is more restricted to the remote area of the south. Water storage containers, especially water jars, served as a main larval breeding habitats of Ae. aegypti, whereas broken cans and plastic containers are considered primary breeding sites for Ae. alpopictus during the dry period. In addition, Aedes larval indices, container index (CI), house index (HI), and Breteau index (BI) were measured. CI and HI values from the central part were significantly higher than those from other areas (p<0.01). BI values of all collection sites were greater than 50 (a maximum BI value accepted by the Ministry of Public Health, Thailand). In brief, Ae. aegypti and Ae. albopictus populations heavily infested many towns and residential areas of the country. Drought could not limit the density of Aedes mosquitos in Thailand. Systematic vector control and vector surveillance programs by public health organizations, if practical, should be continuously conducted to reduce or prevent dengue risk.