Pyrethroid Susceptibility in Stomoxys calcitrans and Stomoxys indicus (Diptera: Muscidae) Collected from Cattle Farms in Southern Thailand.

The susceptibility to six pyrethroid insecticides (permethrin, deltamethrin, alpha-cypermethrin, cypermethrin, lambda-cyhalothrin, and bifenthrin), each at the recommended concentration, was evaluated for two stable fly species-Stomoxys calcitrans (Linnaeus, 1758) and Stomoxys indicus Picard, 1908 (Diptera: Muscidae)-through tarsal contact using a World Health Organization (WHO) cone bioassay procedure. The field populations of S. calcitrans were collected from the Songkhla and Phattalung provinces, while S. indicus were collected from the Phattalung and Satun provinces in Thailand. The stable flies were exposed to insecticide-treated filter paper for 30 min, and their knockdown counts at 30 min and 60 min and mortality counts at 12 h and 24 h were recorded. The S. calcitrans and S. indicus Songkhla and Phattalung populations were moderately susceptible to pyrethroids, as indicated by the 24 h mortality. Nonetheless, the Satun population of S. indicus was completely susceptible to permethrin, with 100% mortality, and showed the lowest susceptibility to deltamethrin and bifenthrin. The results indicate the generally low susceptibility of stable flies to pyrethroids in the southern provinces of Thailand.

UV Light-Emitting-Diode Traps for Collecting Nocturnal Biting Mosquitoes in Urban Bangkok.

Well-designed surveillance systems are required to facilitate a control program for vector-borne diseases. Light traps have long been used to sample large numbers of insect species and are regarded as one of the standard choices for baseline insect surveys. The objective of this study was to evaluate the efficacy of six ultraviolet light-emitting diodes and one fluorescent light for trapping urban nocturnal mosquito species within the Kasetsart University (KU), Bangkok. Ultraviolet light-emitting diodes (UV-LEDs), (LED365, LED375, LED385, LED395, and LED405) and a fluorescent light were randomly assigned to six different locations around the campus in a Latin square design. The traps were operated continuously from 18:00 h to 06:00 h throughout the night. The traps were rotated between six locations for 72 collection-nights during the dry and wet seasons. In total, 6929 adult mosquitoes were caught, with the most predominant genus being Culex, followed by Aedes, Anopheles, Armigeres and Mansonia. Among the Culex species, Culex quinquefasciatus (n = 5121: 73.9%) was the most abundant followed by Culex gelidus (n = 1134: 16.4%) and Culex vishnui (n = 21: 0.3%). Small numbers of Aedes, Armigeres, and Anopheles mosquitoes were trapped [Aedes albopictus (n = 219: 3.2%), Aedes pocilius (n = 137: 2.0%), Armigeres subalbatus (n = 97: 1.4%), Anopheles vagus (n = 70: 1.0%), Aedes aegypti (n = 23: 0.3%)]. There were 2582 specimens (37.2%) captured in fluorescent light traps, whereas 942 (13.6%), 934 (13.5%), 854 (12.3%), 820 (11.8%), and 797 (11.5%) were captured in the LED375, LED405, LED395, LED365, and LED385 traps, respectively. None of the UV-LED light traps were as efficacious for sampling nocturnal mosquito species as the fluorescent light trap. Among the five UV-LED light sources, LED375 trapped the greatest number of mosquitoes. Additional field trials are needed to validate these findings in different settings in order to substantially assess the potential of the LEDs to trap outdoor nocturnal mosquitoes.

Comparing Light-Emitting-Diodes Light Traps for Catching Anopheles Mosquitoes in a Forest Setting, Western Thailand.

Light traps are a common method for attracting and collecting arthropods, including disease vectors such as mosquitoes. Various types of traps have been used to monitor mosquitoes in a forest in Western Thailand. In this study, four Light Emitting Diodes (LED) light sources (UV, blue, green, and red) and two fluorescent lights (white and UV) were used to trap nocturnal adult mosquitoes. These traps were used with light alone and not any additional attractant. The experiment was conducted from 18:00 to 06:00 h. on six consecutive nights, every two months, across dry, wet, and cold seasons. All specimens were first identified by morphological features and subsequently confirmed by using PCR. We collected a total of 873 specimens of 31 species in four genera, Anopheles, Aedes, Culex, and Armigeres. Anopheles harrisoni was the predominant species, followed by Aedes albopictus, Culex brevipalpis, Culex nitropunctatus, and Armigeres (Leicesteria) longipalpis. UV fluorescent light was the most effective light source for capturing forest mosquitoes, followed by UV LED, blue LED, green LED, white fluorescent, and red LED. The optimal times for collection were from 21:00 to 03:00 h in the dry season. Our results demonstrate that appropriate sampling times and light sources should be selected for optimal efficiency in vector surveillance programs.

Species diversity and insecticide resistance within the Anopheles hyrcanus group in Ubon Ratchathani Province, Thailand.

BACKGROUND: Members of the Anopheles hyrcanus group have been incriminated as important malaria vectors. This study aims to identify the species and explore the insecticide susceptibility profile within the Anopheles hyrcanus group in Ubon Ratchathani Province, northeastern Thailand where increasing numbers of malaria cases were reported in 2014. METHODS: Between 2013 and 2015, five rounds of mosquito collections were conducted using human landing and cattle bait techniques during both the rainy and dry seasons. Anopheles mosquitoes were morphologically identified and their insecticide susceptibility status was investigated. Synergist bioassays were carried out with An. hyrcanus (s.l.) due to their resistance to all insecticides. An ITS2-PCR assay was conducted to identify to species the Hyrcanus group specimens. RESULTS: Out of 10,361 Anopheles females collected, representing 18 taxa in 2 subgenera, 71.8% were morphologically identified as belonging to the Hyrcanus Group (subgenus Anopheles), followed by An. barbirostris group (7.9%), An. nivipes (6.5%), An. philippinensis (5.9%) and the other 14 Anopheles species. Specimens of the Hyrcanus Group were more prevalent during the rainy season and were found to be highly zoophilic. Anopheles hyrcanus (s.l.) was active throughout the night, with an early peak of activity between 18:00 h and 21:00 h. ITS2-PCR assay conducted on 603 DNA samples from specimens within the Hyrcanus Group showed the presence of five sisters species. Anopheles peditaeniatus was the most abundant species (90.5%, n = 546), followed by An. nitidus (4.5%, n = 27), An. nigerrimus (4.3%, n = 26), An. argyropus (0.5%, n = 3), and An. sinensis (0.2%, n = 1). All An. hyrcanus (s.l.) specimens that were found resistant to insecticides (deltamethrin 0.05%, permethrin 0.75% and DDT 4% and synergist tests) belonged to An. peditaeniatus. The degree of resistance in An. peditaeniatus to each of these three insecticides was approximately 50%. Addition of PBO (Piperonyl butoxide), but not DEF (S.S.S-tributyl phosphotritioate), seemed to restore susceptibility, indicating a potential role of oxidases as a detoxifying enzyme resistance mechanism. CONCLUSIONS: A better understanding of mosquito diversity related to host preference, biting activity and insecticide resistance status will facilitate the implementation of locally adapted vector control strategies.

Plasmodium Infections in Anopheles Mosquitoes in Ubon Ratchathani Province, Northeastern Thailand During a Malaria Outbreak.

An unprecedented malaria outbreak occurred in Ubon Ratchathani Province, northeastern Thailand, in 2014. The province showed the highest number of malaria cases of all Thai provinces. Five entomological surveys were conducted at 8 sentinel sites from September 2013 to September 2015 to address the role of different Anopheles species in malaria transmission. Mosquito collections were conducted using human landing catches and cow bait. A total of 10,369 Anopheles mosquitoes were collected and 2,240 were morphologically identified as potential malaria vectors, including An. dirus (n = 78), An. minimus (n = 18), An. sawadwongporni (n = 4), An. barbirostris s.l. (n = 819), An. philippinensis (n = 612), An. nivipes (n = 676), An. annularis (n = 42), An. aconitus (n = 7), and An. rampae (n = 142). Real-time polymerase chain reaction was used to screen for the presence of Plasmodium spp. in salivary glands. The proportion of primary vectors of surveyed villages was very low (<1%), and no Plasmodium-infected specimens were detected among in the 2,240 Anopheles mosquitoes tested. The absence of positive Plasmodium samples during malaria outbreaks suggests that malaria transmission most likely occurred outside the villages, particularly in the deep-forested hilly areas that provided suitable habitats for competent malaria vectors. These results emphasize the need to develop vector control related to village community activities to reduce malaria transmission along Thailand border areas.

Abundance and distribution of Anopheles mosquitoes in a malaria endemic area along the Thai-Lao border.

Malaria is an important public health problem in Thailand, especially along international borders. In this study, we conducted a longitudinal entomological survey in six villages and rubber plantation sites to address the spatio-temporal abundance and behavior of malaria vectors in Ubon Ratchathani Province along the Thailand-Laos border. Adult female mosquitoes were collected by human landing collections (indoor and outdoor) and by cattle bait collections twice per year, during rainy and dry seasons. Mosquitoes were morphologically identified and sibling species were determined by allele-specific PCR. Of the 10,024 Anopheles, 9,328 (93.1%) and 696 (6.9%) were collected during the rainy and dry seasons, respectively. A total of 9,769 (97.5%) and 255 (2.5%) was collected on cattle and human baits, respectively. Very few primary and secondary malaria vectors were collected, consisting of 12 specimens of An. dirus, eight An. minimus, and seven An. aconitus. Of the 152 specimens of the Maculatus Group, only three were identified to An sawadwongporni by molecular methods. The others were 112 An. rampae, a non-vector, that were not amplified or were misidentified as other non-vectors. The very low density of primary malaria vectors found in the study villages suggests that entomological risk and malaria transmission is higher in neighboring forest areas. Further studies on malaria vector distribution, as well as human behaviors, are needed to understand malaria transmission dynamics in the province and to develop suitable vector control methods.

Status of insecticide resistance in Anopheles mosquitoes in Ubon Ratchathani province, Northeastern Thailand.

BACKGROUND: Malaria is common in hilly, forested areas along national borders in Southeast Asia. Insecticide resistance in malaria vectors has been detected in a few countries in the Greater Mekong sub-region (GMS), representing a threat to malaria control and prevention. This study aims to determine the insecticide resistance status of Anopheles mosquitoes in Ubon Ratchathani province, northeastern Thailand, where increasing number of malaria cases were reported recently. METHODS: Mosquitoes were collected in 2013-2015 using human landing and cattle bait collections in six sites during both the rainy and dry seasons. Mosquitoes were first morphologically identified to species and their susceptibility status to deltamethrin (0.05%), permethrin (0.75%) and DDT (4%) investigated, according to WHO guidelines. Bioassays with the synergists PBO and DEF were carried out to address the role of detoxifying enzymes in insecticide resistance. DNA sequencing of a fragment of the voltage-gated sodium channel gene was carried out to detect knock-down resistance (kdr) substitutions at position 1014 in resistant species. RESULTS: Due to low vector abundance, complete bioassays (n ≥ 100 mosquitoes) were only achieved for Anopheles hyrcanus s.l., which was resistant to all insecticides tested (mortality ranged from 45 to 87%). Suspected resistance to DDT was found in Anopheles barbirostris s.l. (mortality 69%), but it was susceptible to deltamethrin (mortality 97-100%) and permethrin (mortality 100%). Although insufficient number of primary vectors were collected, results showed that Anopheles dirus s.l. and Anopheles maculatus s.l. were susceptible to deltamethrin (mortality 100%). Anopheles nivipes and Anopheles philippinensis were susceptible to all three insecticides. PBO significantly increased mortality to deltamethrin and permethrin in pyrethroid-resistant An. hyrcanus s.l. None of the sequenced specimens presented the L1014F or L1014S mutation. DISCUSSION: This study shows that insecticide resistance is present in potential malaria vectors in northeastern Thailand. The absence of kdr mutations in all Anopheles species tested suggests that metabolic resistance is the main mechanism of pyrethroid resistance. This study provides new findings about insecticide susceptibility status of potential malaria vectors in northeastern Thailand that are deemed important to guide malaria vector control.

Bioaccumulation and biosorption of Cd(2+) and Zn(2+) by bacteria isolated from a zinc mine in Thailand.

The three bacteria, Tsukamurella paurometabola A155, Pseudomonas aeruginosa B237, and Cupriavidus taiwanensis E324, were isolated from soils collected from a zinc mine in Tak Province, Thailand. Among these bacteria, P. aeruginosa B237 and C. taiwanensis E324 were tolerant of both cadmium and zinc, while T. paurometabola A155 was highly tolerant of zinc only. Bioaccumulation experiment revealed that Cd(2+) and Zn(2+) were mainly adsorbed on the cell walls of these bacteria rather than accumulated inside the cells. During Cd(2+) and Zn(2+) biosorption, P. aeruginosa B237 and T. paurometabola A155 showed the highest removal efficiencies for Cd(2+) and Zn(2+), respectively. The maximum biosorption capacities of P. aeruginosa B237 and T. paurometabola A155 biomasses for Cd(2+) and Zn(2+) biosorptions were 16.89 and 16.75 mg g(-1), respectively, under optimal conditions. The experimental data of Cd(2+) and Zn(2+) biosorptions fitted well with Langmuir isotherm model, suggesting that Cd(2+) and Zn(2+) adsorptions occurred in a monolayer pattern on a homogeneous surface. Furthermore, the pseudo-second order and pseudo-first order kinetic models best described the biosorption kinetics of Cd(2+) and Zn(2+) adsorptions, respectively, suggesting that the Cd(2+) and Zn(2+) adsorptions took place mainly by chemisorption (Cd(2+)) and physisorption (Zn(2+)).