Life table of forensically important blow fly, Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae).

The life table of the Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) were studied under the laboratory conditions at 28 ± 2.5ºC and 75 ± 10% relative humidity using field strain from Penang, Malaysia. The raw data collected were analysed based on the age-stage, two-sex life table theory in order to consider both sexes and the variable developmental rate among individuals and between sexes. The population parameters; intrinsic rate of natural increase (r) was 0.2361 d-1, the finite rate of increase (λ) was 1.2663 d-1, the net reproduction rate (R0) was 97.2 and the mean generation time (T) was 19.4. The two-sex life table analysis provides a comprehensive description of the changes in structure stage of C. rufifacies which is potentially useful for forensic investigations in the country.

Surveillance of Aedes albopictus Skuse breeding preference in selected dengue outbreak localities, peninsular Malaysia.

Entomological surveillance was conducted in order to determine the abundance and to evaluate any changes of biological vectors or ecology, especially in the dengue outbreak areas. The abundance and breeding preference of Aedes albopictus and Aedes aegypti were conducted in selected dengue outbreak localities in three states of peninsular Malaysia namely Selangor, Federal Territory of Kuala Lumpur, and Penang Island using ovitraps and larval survey method. It was determined that Ae. albopictus was predominant in most of the localities and found to breed more outdoor than indoor. A wide range of breeding foci were recorded in this study. It was also determined that ovitrap method was more effective to detect the presence of Aedes mosquitoes when the larval survey was at low rate of infestation. The abundance of Ae. albopictus in dengue outbreak localities emphasis that the vector control programme should also target this species together with the primary dengue vector, Ae. aegypti.

Species composition and pyrethroid susceptibility status of Anopheles mosquitoes from two different locations in Malaysia.

A 14-months survey was carried out to identify the species composition of Anopheles mosquitoes from Kampung Bongor, Grik, Perak. Adding to that, a preliminary one month mosquito population screening was done at Kampung Tepin, Serian, Sarawak. Consequently, the insecticide susceptibility status of a pyrethroid was tested against two selected species of Anopheles collected from these two locations in Malaysia. A total of 4,497 Anopheles from 11 species were identified from collections in Kampung Bongor, whereas 2,654 An. letifer were collected from Kampung Tepin. The An. maculatus of Kampung Bongor and An. letifer of Kampung Tepin were then selected and tested using WHO standard diagnostic test kits and impregnated papers with 0.75% permethrin. The response values of KT50 and KT95 for An. maculatus were recorded at 28.09 minutes and 62.98 minutes respectively. Anopheles letifer recorded much slower response values of KT50 and KT95, which was at 35.09 minutes and 73.03 minutes respectively. Both An. maculatus and An. letifer showed 100% mortality after 24 hours holding period. The results indicate that both species were still susceptible to the tested pyrethroid. For effective vector control and resistance management, accurate and periodic insecticide resistance monitoring should be undertaken especially in rural areas with agricultural usage of insecticides.

Species composition and pyrethroid susceptibility status of Anopheles mosquitoes from two different locations in Malaysia

A 14-months survey was carried out to identify the species composition of Anopheles mosquitoes from Kampung Bongor, Grik, Perak. Adding to that, a preliminary one month mosquito population screening was done at Kampung Tepin, Serian, Sarawak. Consequently, the insecticide susceptibility status of a pyrethroid was tested against two selected species of Anopheles collected from these two locations in Malaysia. A total of 4,497 Anopheles from 11 species were identified from collections in Kampung Bongor, whereas 2,654 An. letifer were collected from Kampung Tepin. The An. maculatus of Kampung Bongor and An. letifer of Kampung Tepin were then selected and tested using WHO standard diagnostic test kits and impregnated papers with 0.75% permethrin. The response values of KT50 and KT95 for An. maculatus were recorded at 28.09 minutes and 62.98 minutes respectively. Anopheles letifer recorded much slower response values of KT50 and KT95, which was at 35.09 minutes and 73.03 minutes respectively. Both An. maculatus and An. letifer showed 100% mortality after 24 hours holding period. The results indicate that both species were still susceptible to the tested pyrethroid. For effective vector control and resistance management, accurate and periodic insecticide resistance monitoring should be undertaken especially in rural areas with agricultural usage of insecticides.

Assessing the susceptibility status of Aedes albopictus on Penang Island using two different assays.

Routine surveillance on resistant status of field mosquito populations is important to implement suitable strategies in order to prevent pest outbreaks. WHO test kit bioassay is the most frequent bioassay used to investigate the susceptibility status of field-collected mosquitoes, as it is relatively convenient to be carried out in the field. In contrast, the topical application of active ingredient is less popular in investigating the susceptibility status of mosquitoes. In this study, we accessed the susceptibility status of Aedes albopictus Skuse collected from two dengue hotspots on Penang Island: Sungai Dua and Persiaran Mayang Pasir. Two active ingredients: permethrin and deltamethrin, were used. WHO test kit bioassay showed that both wild strains collected were susceptible to the two active ingredients; while topical application assay showed that they were resistant. This indicated that WHO test kit bioassay less sensitive to low level of resistance compared to topical application assay. Hence, topical application is expected to be more indicative when used in a resistance surveillance programme.

Entomological study of chikungunya infections in the State of Kelantan, Malaysia.

BACKGROUND & OBJECTIVES: Chikungunya infection has become a public health threat in Malaysia since the 2008 nationwide outbreaks. Aedes albopictus Skuse has been identified as the chikungunya vector in Johor State during the outbreaks. In 2009, several outbreaks had been reported in the State of Kelantan. Entomological studies were conducted in Kelantan in four districts, namely Jeli, Tumpat, Pasir Mas and Tanah Merah to identify the vector responsible for the virus transmission. METHODS: CHIKV cases records were obtained from State Health Department, Kelantan and localities involved were identified. Larva survey was conducted to collect the immature mosquito stages. Modified aspirators were used to collect the adult mosquitoes. All samples on dry ice were transferred to laboratory and the presence of the virus was detected using reverse transcriptase PCR. RESULTS: A total of 1,245 mosquito larvae were collected during larval survey and 2,019 adult mosquitoes were collected using aspirator. From these collections, 640 mosquito pools were tested for the presence of CHIKV by RT-PCR but none found positive. Ae. albopictus was the most abundant mosquito collected, followed by Culex sp., Armigeres sp. and Anopheles sp. A total of 2, 814 artificial containers were inspected during the study. INTERPRETATION & CONCLUSIONS: Since none of the mosquito samples was found to be positive for chikungunya virus, the vector(s) of chikungunya virus in these localities could not be identified.

Assessing the susceptibility status of Aedes albopictus on Penang Island using two different assays

Routine surveillance on resistant status of field mosquito populations is important to implement suitable strategies in order to prevent pest outbreaks. WHO test kit bioassay is the most frequent bioassay used to investigate the susceptibility status of field–collected mosquitoes, as it is relatively convenient to be carried out in the field. In contrast, the topical application of active ingredient is less popular in investigating the susceptibility status of mosquitoes. In this study, we accessed the susceptibility status of Aedes albopictus Skuse collected from two dengue hotspots on Penang Island: Sungai Dua and Persiaran Mayang Pasir. Two active ingredients: permethrin and deltamethrin, were used. WHO test kit bioassay showed that both wild strains collected were susceptible to the two active ingredients; while topical application assay showed that they were resistant. This indicated that WHO test kit bioassay less sensitive to low level of resistance compared to topical application assay. Hence, topical application is expected to be more indicative when used in a resistance surveillance programme.

Temporal fluctuations of insecticides resistance in Musca domestica Linn (Diptera: Muscidae) in Malaysia.

House flies were collected from April 2007-April 2008 from two poultry farms (Balik Pulau and Juru) in the state of Penang. The resistance level of the first generation offspring was evaluated against DDT, malathion, propoxur, and permethrin using the topical application method. The resistance ratio (RR) of the Balik Pulau strain house flies for propoxur, malathion and DDT ranged from 10.28 to 99.00, 7.83 to 47.01 and 6.05 to 31.10, respectively. Resistance to propoxur and malathion in house fly was attributed to cross resistance to organophosphate insecticides used in the farm. Increased metabolic detoxification might be the mechanism involved in DDT resistance due to excessive application of cypermethrin formulation. The RR of the Juru strain for propoxur, malathion and DDT was in a decreasing pattern throughout the study period, ranging from 5.58 to 83.38, 15.19 to 27.82, and 10.04 to 22.69, respectively. Permethrin appeared to be the most potent insecticide in controlling house fly in both the Balik Pulau (RR=0.50 to 1.96) and Juru poultry farms (RR=0.64 to 2.40). The fluctuations of insecticides resistance in house fly was also found to correlate with climatic factors due to its rapid breeding. Relative humidity exhibited positive correlation indices with the changes in the resistance level for DDT (r=0.481, p<0.05), malathion (r=0.698, p<0.01), and permethrin (r=0.580, p<0.05) in Balik Pulau. Similarly, relative humidity in Juru also showed strong correlation with the RR for DDT (r=0.900, p<0.01), malathion (r=0.762, p<0.05), permethrin (r=0.760, p<0.05), and propoxur (r=0.897, p<0.01).

Seasonal abundance of Aedes albopictus in selected urban and suburban areas in Penang, Malaysia.

Ovitrap surveillance was conducted in a selected urban area and suburban area, ie. Taman Permai Indah(TPI) and Kampung Pasir Gebu (KPG) in Penang for 14 months. It was found that Aedes albopictus was the most abundant Aedes species in both study areas, even though a small percentage of Aedes aegypti and Culex quinquefasciatus were found to breed simultaneously in the same ovitrap. This study indicated that the main dengue vector was Ae. albopictus. A strong correlation was found between rainfall and egg population in both of the study sites (r = 0.982 and r = 0.918).

Field evaluation of Bacillus thuringiensis H-14 against Aedes mosquitoes.

Studies were carried out on the residual efficacy of Bacillus thuringiensis H-14 (water dispersible granule, VectoBac ABG 6511) as direct application in the control of Aedes larvae in the field. Field Aedes sp populations in the earthen and glass jars were predetermined before initiation of the trial. On confirmation of the presence of Aedes species in the designated area, Sungai Nibong Kecil, Penang Island, Malaysia, Bti was introduced in the 55L earthen and 3L glass jars). Two test designs were carried out. The first design had treated water replenished daily with 6L of seasoned water and the second design is without the replenishment of water but evaporated water was replenished. Bti was effective in the field for at least 35 days with more than 80% reduction in the Aedes larvae in the treated containers. For earthen jars with daily replenishment of water, 100% reduction was recorded for the first 3 days, while more than 80% reduction was recorded up to day 40. At day 60, Bti still provided an efficacy of 54.32 +/- 4.61 (%) of reduction. Whilst for earthen jars without daily replenishment of water, 100% reduction was recorded for the first 5 days, while more than 80% of reduction was recorded up to day 40. For the glass jars studied, similar efficacy was observed. In jars with daily replenishment of water a better larval control was observed. Percentage of reduction from day 50 to 60 for replenishment of water was between 50 to 70% compared to without replenishment of water with less than 40%.

Susceptibility of laboratory and field-collected Aedes aegypti and Aedes albopictus to Bacillus thuringiensis israelensis H-14.

Susceptibility levels of a few laboratory-cultured and dengue-endemic area field-collected strains of Aedes aegypti and Aedes albopictus to Bacillus thuringiensis israelensis (Bti) at different storage ages were studied. The susceptibility of laboratory-cultured World Health Organization (WHO) Bora Bora reference, Vector Control Research Unit (VCRU), and Fumakilla Malaysia Berhad (FMB) strains of Ae. aegypti to Bti was examined. The sensitivity to Bti decreased with storage age. The median lethal concentration (LC50) for Bti increased by 2-3 times after 2 years compared to a fresh sample (3-6 months of storage). However, after the 2-year storage period, Bti still provided very good efficacy against all laboratory-cultured susceptible strains of Ae. aegypti and Ae. albopictus. The observed 95% lethal concentration values were about 20 times lower than the recommended concentration (6,000 international toxic units (ITU)/liter). Results obtained from the study against the dengue-endemic area field-collected strains of Ae. aegypti and Ae. albopictus confirmed the effectiveness of the Bti after storage for 2 years (18-24 months). For Ae. aegypti, the Ujung Batu strain was the most susceptible to Bti, whereas the Sungai Nibong strain showed the most tolerance. Susceptibility of laboratory-cultured strains varied; the Air Itam strain of Ae. albopictus was the most susceptible to Bti, whereas the Kampung Serani strain was the most tolerant among the field strains. However, the laboratory strain of Ae. albopictus was more susceptible than all the field strains.

Integration of Bacillus thuringiensis H-14 formulations and pyriproxyfen for the control of larvae of Aedes aegypti and Aedes albopictus.

Studies were carried out on the bioefficacy and residual activity of Bacillus thuringiensis israelensis H-14 (Bti) (water-dispersible granules of VectoBac ABG 6511 and liquid formulations of VectoBac 12AS) and pyriproxyfen (insect growth regulator, Sumilarv 0.5%) as direct applications for control of larvae of Aedes aegypti and Aedes albopictus. Two dosages of each Bti formulation (285 and 570 international toxic units [ITU]/liter) and the integration of both Bti formulations and pyriproxyfen were used for residual tests with 45-liter earthen jars for a period of 4 wk. In 1 test series, the treated water was replenished daily with 6 liters of seasoned untreated water. In the 2nd test series, the water in the jars was topped up to the 40-liter level during evaluation. Neither Bti formulation remained effective for a full week. Water-dispersible Bti granules provided effective initial control activity against Ae. aegypti and Ae. albopictus for both test designs (with replenishment and without replenishment of water). The higher dosage (570 ITU/liter) for both Bti formulations was only partially effective at the end of 1 wk after being diluted. After 1 wk, water-dispersible Bti granules provided greater larval mortality than did liquid Bti formulation against both mosquito species when integrated with pyriproxyfen. Pyriproxyfen (79.5 and 159 mg/liter) on its own showed low larvicidal activity but provided very effective control of adult emergence. In this study, integration of Bti (285 and 570 ITU/liter) with pyriproxyfen (79.5 mg/liter) extended the duration of partial larval control somewhat, but live larvae persisted throughout the 4-wk test. The integration effect was more obvious when water-dispersible Bti granules were integrated with pyriproxyfen than when liquid Bti was used. Integration of Bti with pyriproxyfen had a negative effect on adult emergence, which was completely inhibited by pyriproxyfen after day 1. Daily replenishment of water increased Bti activity and provided slightly better larval control. Aedes albopictus and Ae. aegypti were both completely susceptible to the higher concentration of Bti and pyriproxyfen in both test designs (with replenishment and without replenishment of water).

Laboratory evaluation of Bacillus thuringiensis H-14 against Aedes aegypti.

Laboratory efficacy and residual activity of a water dispersible granule formulation of Bacillus thuringiensis israelensis (Bti) at the dosages of 3000, 6000 and 15000 ITU/L were conducted in this study. The study was conducted in two different size containers, earthen jar (45 L) and glass jar (3 L) with or without daily replenishment of 6 L and 0.3 L of water in the earthen and glass jars, respectively. Results indicate that for both earthen jar and glass jar evaluations, Bti at the tested dosages, performed effectively against Aedes aegypti, giving a minimum of 42 days effective killing activity. When the dosage was increased from 3000 ITU/L to 6000 ITU/L or 15000 ITU/L, the effective periods of the Bti increased by an additional one to three weeks. The Bti water dispersible granule provided better larvicidal activity with replenishment of water compared with non-replenishment of water especially for the higher dosage (15000 ITU/L).

Indoor thermal fogging against vector mosquitoes with two Bacillus thuringiensis israelensis formulations, Vectobac ABG 6511 water-dispersible granules and Vectobac 12AS liquid.

Bioefficacy of thermal fogging application of 2 Bacillus thuringiensis israelensis formulations, Vectobac ABG 6511 water-dispersible granules (3,000 international toxic unit [ITU]/mg) and Vectobac 12AS liquid (1,200 ITU/mg), was assessed for larvicidal activities against Aedes aegypti, Aedes albopictus, Anopheles dirus, and Culex quinquefasciatus. Portable Agrofog AF35 sprayers were used to apply the 2 formulations indoors in residential premises on Penang Island, Malaysia. Vectobac ABG 6511 showed good larvicidal effect against all 4 mosquito species at 3 of the higher doses tested (2.91 x 10(9), 1.45 x 10(9), and 0.71 x 10(9) ITU/ha), with more than 96% mortality at 48 h after spraying. As a comparative formulation, Vectobac 12AS also showed good larvicidal activity against all 4 mosquito species at 2 of the higher doses tested (2.87 x 10(9) and 1.46 x 10(9) ITU/ha), with more than 92.5% mortality at 48 h after spraying. Larvae of An. dirus were significantly more susceptible to both water-based Vectobac formulations when compared to the other 3 mosquito species. Both microbial formulations showed better efficacy at higher doses. However, even at the lowest dose tested, Vectobac ABG 6511 and Vectobac 12AS (both at 0.36 x 10(9) ITU/ha) showed larvicidal properties, with more than 66% mortality at 48 h after spraying. Overall, for this bacterial agent, the water-dispersible granule formulation has better prospects than the liquid formulation for the control of larvae of vector mosquitoes.

Indoor thermal fogging application of pesguard FG 161, a mixture of d-tetramethrin and cyphenothrin, using portable sprayer against vector mosquitoes in the tropical environment.

Indoor bioefficacy of the thermal fogging application of Pesguard FG 161, a formulation containing both knockdown and killing agents (active ingredient [AI]: d-tetramethrin 4% [w/w] and cyphenothrin 12% [w/w]) was compared with Resigen5 (AI: s-bioallethrin 0.8% [w/w], permethrin 125/75] 18.7% [w/w], and piperonyl butoxide 16.8% [w/w]), another pyrethroid formulation, as larvicides and adulticides against Aedes aegypti, Aedes albopictus, Anopheles sinensis, and Culex quinquefasciatus using a portable Agrofog AF35 sprayer indoors in houses on Penang Island, Malaysia. Pesguard FG 161 at the concentrations tested was effective against all 4 mosquito species tested. The water-based Pesguard FG 161 performed far better as a larvicide than the diesel-based formulation. Resigen was also effective as a larvicide and adulticide against all 4 mosquito species tested. Larvae of Ae. aegypti were most susceptible to water-based Pesguard FG 161, followed by Cx. quinquefasciatus, An. sinensis, and Ae. albopictus. Even at the lowest concentrations tested, Pesguard FG 161 showed adequate adulticidal properties. At higher dosages, water-based Pesguard FG 161 proved effective as a larvicide against all 4 mosquito species.

Field efficacy of four insect repellent products against vector mosquitoes in a tropical environment.

Four insect repellent products (RPs) (RP 1, Experimental Repellent Lotion [Bayrepel 12%]; RP 2, Experimental Repellent Cream [Bayrepel 5%]; RP 3, Off! Insect Repellent II Aerosol [deet 15%]; and RP 4, Off! Skintastic II Cream [deet 7.5%]) were evaluated simultaneously for their efficacy against vector and nuisance mosquitoes. The aim of this study was to compare the relative efficacy of RPs based on a new repellent compound, Bayrepel (1-piperidinecarboxylic acid, 2-(2-hydroxyethyl)-1-methylpropylester), with deet (N,N-diethyl-m-toluamide)-based RPs. An 8-h field efficacy of above repellents was evaluated against the day-biting mosquito (Aedes albopictus) and night-biting mosquitoes (Culex quinquefasciatus and Anopheles spp.). Evaluation was carried out by exposing humans with repellent-treated bare limbs to mosquitoes landing and to mosquitoes landing and biting. Repellent product 1 or 2 was applied on the left arm and leg, whereas RP 3 or 4 was applied on the right arm and leg, respectively. Application of these 4 RPs significantly reduced (P < 0.05) the landing and the landing and biting of day-biting and night-biting mosquitoes. All 4 RPs were found to be equally effective (P < 0.05) against Ae. albopictus and Cx. quinquefasciatus. However, for protection against Anopheles spp., RPs 1 and 3 exhibited significantly (P < 0.05) better repellency effect than RPs 2 and 4.