Evaluation of insecticide resistance among Malaysian Aedes albopictus Skuse larvae based on revised diagnostic doses of larvicides

@#The susceptibility levels of Malaysian Aedes albopictus larvae sampled from several agricultural, fogging-free residential and dengue prone residential areas against different larvicides were evaluated using revised diagnostic doses derived from the 2xLC99 values of the reference strain. Upon 24-hour recovery period of WHO larval bioassay, incipient resistance was observed among Ae. albopictus larvae from rubber estates against fenitrothion (96.67% mortality) and permethin (97.00% mortality) while Ae. albopictus larvae from rice cultivation areas were moderately resistant to fenthion (94.33% mortality). Aedes albopictus larvae from dengue prone residential areas developed moderate to high resistance against dichlorodiphenyltrichloroethane (DDT), fenitrothion, fenthion, propoxur and permethrin (79.67% – 97.33% mortality). Moderate to high resistance were also demonstrated among all populations of Ae. albopictus larvae against temephos and chlorpyrifos (63.00% – 97.67% mortality). Except for Ae. albopictus larvae from oil palm plantations, all Ae. albopictus larval populations were also highly resistant to bendiocarb (65.67% – 89.67% mortality). Cross resistance between larvicides from similar and different insecticide classes were also revealed in this study. The use of revised diagnostic doses established from the local reference strain could reduce the possibility of underestimation or overestimation of the insecticide susceptibility status of field strain populations.

Assessment of the susceptibility status of Aedes albopictus (Diptera: Culicidae) from Interior, Sandakan and Tawau divisions of Sabah, Malaysia based on WHO diagnostic doses of larvicides

@# Susceptibility status of Aedes albopictus (Skuse) sampled from residential areas in Interior, Sandakan and Tawau divisions of Sabah, Malaysia, was evaluated based on the WHOrecommended doses of organochlorine and organophosphate larvicides. To determine susceptibility status, larval bioassays were carried out and post 24-hour mortalities based on WHO resistance classifications were adopted. The results demonstrated that Ae. albopictus larvae were resistant toward 5 out of the 8 larvicides tested. Larvae from all populations were resistant against bromophos, fenitrothion, malathion, temephos and dichlorodiphenyltrichloroethane (DDT), with mortalities ranging from 0.00 to 89.33%. Dieldrin, on the other hand, could induce 100.00% mortalities in all populations, followed by fenthion and chlorpyrifos, with mortalities ranging from 97.33 to 100.00% and 81.33 to 100.00% respectively. Despite most populations exhibiting similitude in their resistance status, larvae from Sandakan exhibited the highest resistance level whereas the lowest level was observed in Keningau. In view of the inadequacy of some larvicides in controlling Ae. albopictus in this study, integrated management such as insecticide rotation or combination of interventions is warranted.

Bioefficacy of mosquito mat vaporizers and associated metabolic detoxication mechanisms in Aedes aegypti (Linnaeus) in Selangor, Malaysia: A statewide assessment

@#This study aims to examine the efficacy of mosquito mat vaporizers on Aedes aegypti and their associated metabolic detoxication mechanisms. For this purpose, Aedes aegypti (Linnaeus) was collected from nine districts in Selangor, Malaysia and tested with mosquito vaporizing mat bioassays. The same populations were also subjected to biochemical assays to investigate activities of detoxifying enzymes, namely non-specific esterase (EST), glutathione-S-transferase (GST) and mixed function oxidase (MFO). The efficacy of Ae. aegypti on the active ingredients tested in decreasing order were d- allethrin > dimefluthrin > prallethrin with PBO > prallethrin. The results further indicated significant enhancement mean levels of EST, GST and MFO in pyrethroid-resistant populations. The mortality rate of Ae. aegypti in response to pyrethroid active ingredients was associated with MFO activity, suggesting it is an important detoxification enzyme for the populations tested. In view of the presence of resistance against household insecticide products, pyrethroid efficacy on Ae. aegypti populations needs to be monitored closely to ensure the implementation of an effective vector control program in Malaysia.

Morphological characteristics of microfilariae in blood smears of the common treeshrew Tupaia glis (Mammalia: Scandentia) in Gemas, Negeri Sembilan, Malaysia

@#Some filarial nematodes, such as Wuchereria bancrofti, Brugia malayi, and Brugia timori, cause lymphatic diseases in humans in the tropics, whereas other filarial parasites from wild animals cause zoonotic diseases in humans worldwide. To elucidate the prevalence and diversity of filarial parasites in Malaysia, we investigated the filarial parasites from wild animals in Gemas, Negeri Sembilan. To find adult filarial parasites, we dissected 26 animals, which included five frogs, one skink, one snake, two birds, six common treeshrews, and 11 rats. Then, we examined microfilariae in the blood smears and skin snips obtained from each animal. We found two types of microfilariae in the blood smears of common treeshrews: one was very similar to Malayfilaria sofiani and the other closely resembled Brugia tupaiae. These findings indicate an additional distribution of these filarial parasites in Gemas.

Descriptions of the female, male and mature larva of Simulium contractum Takaoka (Diptera: Simuliidae) from Sulawesi, Indonesia

@#Simulium (Simulium) contractum Takaoka from Sulawesi, Indonesia was known only as the pupa. Its female, male and mature larva are described for the first time. The tentative assignment of this species in the Simulium dumogaense species-group is confirmed by the adult characters including the female and male genitalia. The female and male of this species are similar to those of Simulium (Simulium) tumpaense Takaoka & Roberts but are distinguished by the yellowish femora.

Preliminary dengue vector surveillance in the Sunda Islands, Indonesia: Interchange of breeding habitat preferences of Aedes aegypti and Aedes albopictus

@#Ovitrap surveillance was conducted to determine the infestation patterns of dengue vectors in fourteen study sites across eight provinces located in the Sunda Islands, Indonesia. High ovitrap indices up to 70% and 90% were obtained from indoor and outdoor areas, respectively. Mean numbers of Ae. aegypti and Ae. albopictus larvae ranged from 0.13 to 14.50 and 0.10 to 18.60, respectively. Mixed infestation (<10%) and interchange of breeding habitat preferences of Ae. albopictus and Ae. aegypti were also observed in the present study.

A new species of the Simulium (Simulium) crocinum species-group (Diptera: Simuliidae) from Vietnam

@#A new black fly species, Simulium yukawai, is described on the basis of one female and its pupal exuviae from Vietnam. This new species is placed in the S. crocinum species-group of Simulium (Simulium) (Diptera: Simuliidae). It is characterized in the female by the paraproct covered with 48–50 distinct hairs on its ventral and lateral surfaces, and in the pupa by the frons and most of the thorax bare, gill with six dark filaments in three pairs closely arising from the base, of which the two outer filaments of the dorsal and middle pairs are much longer than other filaments, abdomen with distinct spine-combs on the dorsal surface of segments 7–9, and cocoon wall-pocket shaped, with a large anterolateral window on each side. Taxonomic notes are given to separate this new species from related species. This species is the fourth member of the S. crocinum species-group known from Vietnam.

Single and co-breeding of different mosquito species in fogging-free and dengue risk areas in West Malaysia

@#Ovitrap surveillance was carried out in fifteen localities encompassing foggingfree and dengue risk areas in West Malaysia to determine on the dispersal and prevalence of single and co-breeding of mosquito larvae particularly Aedes. Ovitraps were sited randomly indoors and outdoors within human settlements in all study areas. All the localities exhibited positive ovitraps with single breeding of Ae. albopictus that ranged between 64.29% and 100.00%. These findings indicated Ae. albopictus as the predominant container-breeding species in all study areas. The co-breeding of Ae. aegypti with Ae. albopictus larvae (34 ovitraps), Ae. albopictus with Culex quinquefasciatus larvae (32 ovitraps) as well as Cx. quinquefasciatus with Armigeres subalbatus larvae (1 ovitrap) were also detected in certain study localities. Interestingly, co-breeding of Ae. albopictus with Ar. subalbatus larvae as well as Ae. albopictus with Uranotaenia sp. larvae in Malaysia is reported for the first time in the present study. Better understanding of the co-breeding scenario involving different species of mosquito larvae is needed to ensure the efficacy of vector control actions to be conducted.

Three new species of black flies (Diptera: Simuliidae) from the Lesser Sunda Archipelago, Indonesia

@#Simulium (Gomphostilbia) dhangi sp. nov., S. (G.) sumbaense sp. nov. and S. (Nevermannia) wayani sp. nov. are described from the Lesser Sunda Archipelago, Indonesia. Simulium (G.) sumbaense sp. nov. is placed in the S. varicorne species-group and is characterized by the pupal gill with eight filaments arranged as (1+1+1+1+2)+2 from dorsal to ventral, while S. (G.) dhangi sp. nov., unplaced to group, is characterized by the pupal gill composed of two inflated trunks and four slender filaments all arising basally, and the short larval antenna as long as the stem of the labral fan. Simulium (N.) wayani sp. nov. belongs to the S. ruficorne species-group and is characterized by the female spermatheca with an unsclerotized neck, and pupal gill with four inflated filaments. The number of species of black flies from the archipelago increases from 19 to 22.

Biting behaviour of medically important mosquitoes (Diptera: Culicidae) in Peninsular Malaysia

Human-landing catch (HLC) technique was undertaken in nine selected study sites within Peninsular Malaysia to determine the biting cycle of Aedes, Armigeres, Culex, and Mansonia mosquitoes. HLC was conducted 24 hours on three different nights in a duration of one week at each study site. Aedes albopictus were found in all study sites with bimodal biting cycle, whereas the biting peak of Aedes butleri was recorded between 1600 and 1900 hr. For Armigeres, five study sites demonstrated similar biting peak hour (1900 – 2000 hr) for Ar. kesseli four study sites showed biting peak at 1800 – 1900 hr for Ar. subalbatus. Culex quinquefasciatus was collected from all study sites except in Sungai Penchala with its biting activity beginning from 1900 hr. Both Mansonia annulifera and Mansonia uniformis captured in two different study sites showed similar biting peaks (1500 – 1600 hr). It is recommended that vector control activities be carried out at dusk as almost all mosquito species captured started their biting activities at that time.

Distribution and abundance of black flies (Diptera: Simuliidae) in recreation parks in Selangor State, Peninsular Malaysia

A preliminary survey of larvae and pupae of black flies (Diptera: Simuliidae) was conducted in three recreation parks [Templer Recreation Park (TRP), Congkak River Forest Reserve (CRFR) and Ampang Forest Reserve (AFR)] located in Selangor State, which is located 18 to 35 km from Kuala Lumpur city center, Malaysia. This study was initiated to determine the distribution and abundance of filarial vectors, Simulium spp. A total of 12 species of black flies belonging to three subgenera (Gomphostilbia, Simulium and Nevermannia) were collected. Simulium (Simulium) nobile was incriminated as the most dominant species in all recreation parks. This study is a first report on the distribution and abundance of black flies obtained from recreation parks in Malaysia.

First molecular genotyping of A302S mutation in the gamma aminobutyric acid (GABA) receptor in Aedes albopictus from Malaysia

Given the lack of molecular evidence in altered target-site insecticide resistance mechanism in Aedes albopictus (Skuse) worldwide, the present study aims to detect the presence of A302S mutation in the gene encoding the gamma aminobutyric acid receptor resistant to dieldrin (Rdl) in Ae. albopictus for the first time from its native range of South East Asia, namely Malaysia. World Health Organization (WHO) adult susceptibility bioassay indicated a relatively low level of dieldrin resistance (two-fold) in Ae. albopictus from Petaling Jaya, Selangor. However, PCR-RFLP and direct sequencing methods revealed the presence of the A302S mutation with the predomination of heterozygous genotype (40 out of 82 individuals), followed by the resistant genotype with 11 individuals. This study represents the first fieldevolved instance of A302S mutation in Malaysian insect species.

Evaluation of insect growth regulators, temephos and Bacillus thuringiensis israelensis against Aedes aegypti (L) in plastic containers

The residual activities of 5 insect growth regulators (IGRs) were studied and compared to operational dosage of temephos (1 mg/L) and Bacillus thuringiensis israelensis (Bti) (0.008 mg/L). The IGRs, temephos and Bti were applied into plastic containers containing 5 litres of water. Thirty Aedes aegypti larvae were added into each container weekly. Efficacy of these IGRs were evaluated for effective duration of each dosage and the percentage of emergence inhibition (EI). An end-point of EI/mortality > 50% was considered to be effective. Pyriproxyfen possessed the longest residual activity in both indoor (43 weeks) and outdoor (26 weeks) conditions, followed by temephos (26 weeks in indoor and 16 weeks in outdoor). Although the residual activity of Bti in indoor lasted 8 weeks which was longer than cyromazine and diflubenzuron; however, it was least effective in outdoor which only lasted 2 weeks. This study revealed that pyriproxyfen possessed good residual effect among test IGRs compared to temephos and Bti. The use of IGRs can be an alternative long-term control measure in stagnant water body.

Vertical distribution of Aedes mosquitoes in multiple storey buildings in Selangor and Kuala Lumpur, Malaysia

The aim of the present study was to determine the vertical distribution and abundance of Aedes mosquitoes in multiple storey buildings in Selangor and Kuala Lumpur, Malaysia. Ovitrap surveillance was conducted for 4 continuous weeks in multiple storey buildings in 4 residential areas located in Selangor [Kg. Baiduri (KB)] and Kuala Lumpur [Student Hostel of University of Malaya (UM), Kg. Kerinchi (KK) and Hang Tuah (HT)]. The results implied that Aedes mosquitoes could be found from ground floor to highest floor of multiple storey buildings and data from different elevation did not show significant difference. Ovitrap index for UM, KB, HT and KK ranged from 0 – 29.17%, 0 – 55.56%, 8.33 – 83.33% and 0 – 91.17% respectively. Aedes aegypti and Aedes albopictus were found breeding in HT, KK and KB; while only Ae. albopictus was obtained from UM. The results indicate that the invasion of Aedes mosquitoes in high-rise apartments could facilitate the transmission of dengue virus and new approaches to vector control in this type of residential area should be developed.

Impact of larviciding with a Bacillus thuringiensis israelensis formulation, VectoBac WG, on dengue mosquito vectors in a dengue endemic site in Selangor State, Malaysia.

The field bioefficacy of a wettable granule (WG) formulation of Bacillus thuringiensis israelensis (Bti), VectoBac WG (Bti strain AM65-52) against dengue vectors, Aedes aegypti and Ae albopictus; was evaluated in a suburban residential area (TST) and in a temporary settlement site (KB) in the state of Selangor, Malaysia. Pre-control ovitrap surveillance of the trial sites indicated a high population of both types of Aedes mosquitoes. The populations were monitored continuously by weekly ovitrapping. Bti was sprayed biweekly at a dosage of 500 g/ha by using a mist-blower. The spray application was targeted into outdoor larval habitats. If required, Bti formulation was also applied directly into indoor water-holding containers at 8 g/1,000 l. Based on ovitrap surveillance, a significant reduction in Aedes populations was evident 4 weeks after initiating the first Bti treatment. The ovitrap index (OI) and the larvae density decreased drastically in both trial sites. In TST, the indoor OI was significantly reduced from 57.50 +/- 7.50% to 19.13 +/- 5.49% (p<0.05), while the outdoor OI decreased from 38.89 +/- 11.11% to 15.36 +/- 5.93%. In KB, similarly, the OI was significantly reduced by more than half, from 66.66 +/- 6.67% to 30.26 +/- 2.99% (p< 0.05). In all cases, the reduction in OI was paralleled by reduction in larval density.