The resistance status of riceland Culex vishnui against four major groups of insecticides (i.e., organochlorines, carbamates, organophosphates and pyrethroids) was investigated. Biochemical assays (ESTα, ESTβ, MFO and GST) were also conducted to detect the resistance levels. Throughout a 12-month study period, multiple insecticide resistance was observed in both larvae and adult Cx vishnui. Culex vishnui larvae exhibited low resistance against malathion, temephos and permethrin with resistance ratio (RR) values < 5. In adult bioassay, Cx. vishnui were highly resistant against all tested adulticides with 24h post-treatment mortality < 70%. Correlations between permethrin and malathion resistance, as well as between deltamethrin and cyfluthrin resistance were found in Cx. vishnui. The results indicated that mixed function oxidases activity of Cx. vishnui was the highest compared to ESTα, ESTβ and GST. Spearman rank-order analysis showed that ESTα, ESTβ and GST were involved in multiple resistances in Cx. vishnui. The findings of this study established a baseline of insecticide susceptibility and revealed the effects of agricultural insecticide pressure on the vectors of Japanese encephalitis in Malaysia.
This study reported the ant species that were recovered from monkey carcasses in three different ecological habitats in Malaysia. The study was conducted from 9 May - 10 October 2007, 6 May - 6 August 2008 and 26 May - 14 July 2009 in forested area (Gombak, Selangor), coastal area (Tanjong Sepat, Selangor) and highland area (Bukit Cincin, Pahang), respectively. Monkey carcass was used as a model for human decomposition in this study. A total of 4 replicates were used in each of the study sites. Ants were observed to prey on eggs, larvae, pupae and newly emerged flies. This study found that ant species could be found at all stages of decomposition, indicating that ants were not a significant indicator for faunal succession. However, different species of ants were obtained from monkey carcasses placed in different ecological habitats. Cardiocondyla sp. was only found on carcasses placed in the coastal area; while Pheidole longipes, Hypoponera sp. and Pachycondyla sp. were solely found on carcasses placed in the highland area. On the other hand, Pheidologeton diversus and Paratrechina longicornis were found in several ecological habitats. These data suggests that specific ant species can act as geographic indicators for different ecological habitats in forensic entomology cases in Malaysia.
Larvae of Aedes albopictus obtained from dengue endemic areas in Selangor, Malaysia were evaluated for their susceptibility to operational dosage of temephos (1 mg/L). Larval bioassays were carried out in accordance to modified WHO standard methods. Biochemical microassay of enzymes in Ae. albopictus was conducted to detect the emergence of insecticide resistance and to define the mechanisms involved in temephos resistance. The 50% mortality lethal time (LT50) for Ae. albopictus tested against temephos ranged between 58.65 to 112.50 minutes, with resistance ratio ranging from 0.75 - 1.45. This study addressed the fluctuation of time-related susceptibility status of Ae. albopictus towards insecticide. Significant difference on the weekly enzyme levels of non-specific esterases, mixed function oxidases and glutathione S-transferases was detected (p ≤ 0.05). No significant correlation was found between temephos resistance and enzyme activity (p > 0.05). Only glutathione S-transferases displayed high level of activity, indicating that Ae. albopictus may be resistant to other groups of insecticide. The insensitive acetylcholinesterase was detected in some field collected Ae. albopictus populations, indicating the possibility of emergence of carbamate or other organophosphate resistance in the field populations. Continuous resistance monitoring should be conducted regularly to confirm the efficacy of insecticides for dengue control.
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.
Estimation of post-mortem interval (PMI) is crucial for time of death determination. The advent of DNA-based identification techniques forensic entomology saw the beginning of a proliferation of molecular studies into forensically important Calliphoridae (Diptera). The use of DNA to characterise morphologically indistinguishable immature calliphorids was recognised as a valuable molecular tool with enormous practical utility. The local entomofauna in most cases is important for the examination of entomological evidences. The survey of the local entomofauna has become a fundamental first step in forensic entomological studies, because different geographical distributions, seasonal and environmental factors may influence the decomposition process and the occurrence of different insect species on corpses. In this study, calliphorids were collected from 13 human corpses recovered from indoors, outdoors and aquatic conditions during the post-mortem examination by pathologists from the government hospitals in Malaysia. Only two species, Chrysomya megacephala and Chrysomya rufifacies were recovered from human corpses. DNA sequencing was performed to study the mitochondrial encoded COI gene and to evaluate the suitability of the 1300 base pairs of COI fragments for identification of blow fly species collected from real crime scene. The COI gene from blow fly specimens were sequenced and deposited in GenBank to expand local databases. The sequenced COI gene was useful in identifying calliphorids retrieved from human corpses.
Bioassay test against malathion had been carried out with larval and adult stages of Aedes aegypti. The mosquitoes were under selection pressure against malathion for forty-five consecutive generations. The rate of resistance development was measured by LC(50) and LT(50) values. The larvae and adult females, after subjection to malathion selection for 45 generations, developed high resistance level to malathion, with resistance ratio of 52.7 and 3.24 folds, respectively over control mosquitoes. Cross-resistance towards the same and different groups of insecticides was determined using the F44 and F45 malathion-selected adult females. Insecticides tested were DDT (4.0%), permethrin (0.75%), propoxur (0.1%), fenitrothion (1%), λ-cyhalothrin (0.05%) and cyfluthrin (0.15%). Results indicated that the mosquitoes were highly resistant to DDT and fenitrothion, moderately resistant to propoxur, tolerant to permethrin and λ-cyhalothrin, and very low resistant to cyfluthrin.
Simulium (Gomphostilbia) sofiani sp. nov. is described on the basis of reared adult female, male, pupal and larval specimens collected from Cameron Highlands, Pahang state, Malaysia. This new species is placed in the ceylonicum species-group within the subgenus Gomphostilbia and is easily distinguished from all the related known species by the combination of the following characteristics: an elongate sensory vesicle and yellow hair tuft on the stem vein of the wing in the female, the greater number of large upper-eye facets (15 or 16 vertical columns and 15 or 16 horizontal rows) and almost entirely darkened hind basitarsus in the male, and the gill bearing a long common basal stalk and 8 filaments arranged as [(1+2)+(1+2)] +2 filaments from dorsal to ventral in the pupa.
The susceptibility status of field-collected Aedes aegypti (L.) from a dengue endemic area to Bacillus thuringiensis israelensis (Bti) and temephos was determined. Since August 2007, biweekly ovitrap surveillance (OS) was conducted for 12 mo in 2 sites, A & B, in Shah Alam, Selangor. Site A was treated with a Bti formulation, VectoBac® WG at 500 g/ha, from December 2007 - June 2008 while Site B was subjected to routine dengue vector control activities conducted by the local municipality. Aedes aegypti larvae collected from OS in both sites were bred until F3 and evaluated for their susceptibility. The larvae were pooled according to 3 time periods, which corresponded to Bti treatment phases in site A: August - November 2007 (Bti pre-treatment phase); December 2007 - June 2008 (Bti treatment phase); and July - September 2008 (Bti post-treatment phase). Larvae were bioassayed against Bti or temephos in accordance with WHO standard methods. Larvae collected from Site A was resistant to temephos, while incipient temephos resistant was detected in Site B throughout the study using WHO diagnostic dosage of 0.02 mg/L. The LC50 of temephos ranged between 0.007040 - 0.03799 mg/L throughout the year in both sites. Resistance ratios (LC50) indicated that temephos resistance increased with time, from 1.2 - 6.7 folds. The LC50 of Ae. aegypti larvae to Bti ranged between 0.08890 - 0.1814 mg/L throughout the year in both sites, showing uniform susceptibility of field larvae to Bti, in spite of Site A receiving 18 Bti treatments over a period of 7 mo. No cross-resistance of Ae. aegypti larvae from temephos to Bti was detected.
Aedes albopictus was bioassayed to determine resistance development to malathion (OP). Two methods were applied, including WHO larval bioassay to determine the susceptibility to lethal concentration (LC), and adult bioassay to determine lethal time (LT). Larvae from colonies that had undergone selection pressure with malathion to yield 50% mortality were further subjected to selection for subsequent 10 generations. Selection of Ae. albopictus with malathion could relatively induce a consistent resistance ratio of 1.0 throughout 10 generations. It was noted that Ae. albopictus larvae showed less susceptibility to malathion compared to adults. The susceptibility test of adult mosquitoes to diagnostic dosage of 5.0% malathion-impregnated paper showed a variety of susceptibility to malathion when compared to the susceptible strain. Bioassay results indicated that the LT50 values of malathion-selected Ae. albopictus ranged between 11.5 - 58.8 minutes for ten consecutive generations. Biochemical enzyme studies indicated that there was a significant difference (p < 0.05) in esterase level in malathion-selected mosquitoes compared to non-selected control. Electrophoretic patterns of non-specific esterases at different life stages in malathion-selected Ae. albopictus suggested that non-specific esterases do not play a role in resistance of malathion-selected Ae. albopictus.
A preliminary study on the vertical dispersal of Aedes populations in high-rise apartments was carried out in Presint 9, Putrajaya, Malaysia. Ovitraps were placed indoors within four blocks of high-rise apartments from the ground floors (0.0 - 3.0 m) until up to the tenth floors (28.1 - 30.0 m). Aedes aegypti was the dominant species found in the ovitraps (87.85%), while Aedes albopictus was found in lower numbers. From total number of larvae collected (650), 40.92% of these larvae were obtained from the fourth block; Block D. The peak density of Aedes sp. was observed at level 6 (16.1 - 18.0 m), while Ae. aegypti was found until the tenth floor (28.1 - 30.0 m). In contrast, Ae. albopictus was found only up to the sixth floor (16.1 - 18.0 m). A poor correlation of the mean number of Aedes larvae collected with the level of high-rise apartments occupied (N=40; ρ=-0.349) was also observed which indicated the possibility of lesser Aedes populations to be found at higher level of high-rise apartments. Therefore, larger scale studies are strongly recommended to examine the vertical dispersal of Aedes mosquitoes.
A forensic entomological study was conducted using monkey carcasses (Macaca fascicularis Raffles) that were placed in either an outdoor or indoor environment at a coastal area in Tanjung Sepat, Selangor, Malaysia during May until August 2008. We collected pupae of Chrysomya rufifacies (Marquart) from the carcasses and kept them individually. The emergence of 13 parasitic microhymenopteran, from one of the pupae occurring within a week were identified as Exoristobia philippinensis Ashmead (Hymenoptera: Encyrtidae). Another observation was made whereby a pupa of C. rufifacies was predated by a muscid larva, Ophyra spinigera (Stein). The larva squeezed into the pupa and consumed the contents. This paper report C. rufifacies as a new host record for E. philippinensis in Malaysia and highlighted the predatory behavior of O. spinigera larva in natural environment.
A study on insect succession of monkey carcass in a forested area in Ulu Gombak, Selangor, Malaysia was conducted from 9 May to 18 June 2007. The third instar of the housefly, Musca domestica (Linnaeus) (Diptera: Muscidae) were only found on dry stage of a decomposed (Day-33) monkey carcass (Macaca fascicularis Raffles). This observation revealed that M. domestica maggots were found together with other muscid fly maggots, Hydrotaea (=Ophyra) spinigera (Stein) (Diptera: Muscidae) on dry stage of a carcass. However, the role of M. domestica on forensic entomological study remains unknown. This study recorded the first finding of M. domestica maggots on primate carcass in Malaysia.
Signal fly, Scholastes sp. (Diptera: Platystomatidae) was observed associated with animal carcasses in Malaysia. The first observation was on a monkey carcass, which was killed by using a handgun and immediately placed in a forested area in Gombak, Selangor while the second observation was on a pig that died of natural causes and whose carcass was placed in an oil palm plantation in Tanjung Sepat, Selangor. Both animal carcasses were visited by Scholastes sp. flies during the fresh decomposition period. However, the role Scholastes flies in the decomposition process remains unknown. In this paper, we report the occurrence of Scholastes sp. on animal carcasses in Malaysia for the first time.
Ovitrap surveillance was initiated for eight continuous weeks to determine the distribution and abundance of Aedes sp. mosquitoes in the University of Malaya campus, Kuala Lumpur, and the impact of meteorological conditions on the Aedes populations. Two study areas within the campus were selected: Varsity Lake and Seventh Residential College. The abundance of Aedes populations in Varsity Lake was indicated by ovitrap index (OI) which ranged from 60.00%-90.00%. The mean number of larvae per ovitrap of Aedes albopictus in Varsity Lake ranged from 11.23+/-2.42-43.80+/-6.22. On the other hand, the outdoor OI for Seventh Residential College ranged from 73.33%-93.33%, respectively, while the mean number larvae per ovitrap for this area ranged from 19.33+/-4.55-35.27+/-5.46, respectively. In addition, the indoor OI of Seventh Residential College ranged from 0.00%-30.00%, while the mean number of larvae per ovitrap for Ae. albopictus ranged from 0-5.90+/-3.55. There was no significant difference (p>0.05) of Ae. albopictus population between Varsity Lake and Seventh Residential College. The studies showed a correlation between OI and mean number of larvae per ovitrap for outdoor Ae. albopictus populations in Varsity Lake and Seventh Residential College (r=0.794). There was also a correlation between the mean larvae number per ovitrap of Ae. albopictus obtained from eight weeks indoor ovitrap surveillance in Seventh Residential College with rainfall (r=0.584). However, there was no correlation between the mean larvae number per ovitrap of Ae. albopictus in both study areas with temperature and relative humidity. Aedes aegypti mosquitoes were found neither indoor nor outdoor in both study areas. This study indicated that the principal dengue vector in the university campus was most likely Ae. albopictus.
The bioefficacy of nine commercial formulations of temephos against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus larvae were evaluated in the laboratory. WHO larval bioassay with operational dosage of temephos at 1 mg/L was performed. The larval mortality was recorded every 5 minutes until complete mortality was achieved. All formulations of temephos exhibited various toxicity level against Ae. aegypti, Ae. albopictus and Cx. quinquefasciatus. Generally, larvae of Cx. quinquefasciatus was susceptible to all formulations of temephos, followed by Ae. aegypti and Ae. albopictus.
Larvae obtained from Taman Samudera (Gombak, Selangor), Kampung Banjar (Gombak, Selangor), Taman Lembah Maju (Cheras, Kuala Lumpur) and Kampung Baru (City centre, Kuala Lumpur) were bioassayed with diagnostic dosage (0.012 mg/L) and operational dosage (1 mg/L) of temephos. All strains of Aedes aegypti and Aedes albopictus showed percentage mortality in the range of 16.00 to 59.05 and 6.4 to 59.50 respectively, after 24 hours. LT50 values for the 6 strains of Ae. aegypti and Ae. albopictus were between 41.25 to 54.42 minutes and 52.67 to 141.76 minutes respectively, and the resistance ratio for both Aedes species were in the range of 0.68 to 1.82 when tested with operational dosage, 1 mg/L temephos. These results indicate that Aedes mosquitoes have developed some degree of resistance. However, complete mortality for all strains were achieved after 24 hours when tested against 1 mg/L temephos.
Larvae of Aedes aegypti and Aedes albopictus obtained from 6 consecutive ovitrap surveillance (OS) in Taman Samudera and Kg. Banjar were evaluated for their susceptibility to temephos. Larval bioassays were carried out in accordance with WHO standard methods, with diagnostic dosage (0.012 mg/L) and operational dosage (1 mg/L) of temephos respectively. Aedes aegypti and Ae. albopictus obtained from six OS in Taman Samudera showed resistance to diagnostic dosage of temephos with percentage mortality between 5.3 to 72.0 and 9.3 to 56.0, respectively, while Ae. aegypti and Ae. albopictus obtained from Kg. Banjar showed resistance to temephos with percentage mortality between 16.0 to 72.0 and 0 to 50.6, respectively. Only two strains of Ae. aegypti from Kg. Banjar were susceptible to temephos with 93.3% (OS 2) and 100% (OS 3) mortality. The 50% mortality at lethal time (LT50) for all strains of Ae. aegypti and Ae. albopictus tested against operational dosage of temephos showed range between 36.07 to 75.69 minutes and 58.65 to 112.50 minutes, respectively, and complete mortality was achieved after 24 hours. Our results indicated that there is weekly variations of the resistance status for Ae. aegypti and Ae. albopictus. Aedes susceptibility to temephos is changing from time to time in these two study sites. It is essential to continue monitoring the resistance of this vector to insecticides in order to ensure the efficiency of program aimed at vector control and protection of human health.
Ovitrap surveillance was conducted in four dengue endemic areas in Kuala Lumpur and Selangor, Malaysia to determine the distribution and percentage of mixed breeding of both Aedes aegypti and Aedes albopictus. The percentage of mixed breeding in all study sites both indoors and outdoors accounted for 10 to 32 % from the total ovitraps collected. Ae. aegypti was found at a higher frequency than Ae. albopictus in these ovitraps. This study again indicates that ovitrap is a sensitive tool to attract gravid females of more than one mosquito species to oviposit in the container.
Laboratory-bred females of Culex quinquefasciatus, Aedes aegypti and Aedes albopictus from the insectarium, Unit of Medical Entomology, Institute for Medical Research were used in the experiment. The late third stage of the F0 larvae which survived the high selection pressure of malathion, permethrin and temephos were reared and colonies were established from adults that emerged. Cx. quinquefasciatus larvae were subjected to selection by malathion and permethrin for 40 generations, Ae. aegypti larvae to malathion, permethrin and temephos for 32 generations and Ae. albopictus larvae were selected against malathion and permethrin for 32 generations and 20 generations against temephos. The rate of resistance development was measured by LC50 value. Cx. quinquefasciatus larvae developed higher resistance to malathion and permethrin compared to Ae. aegypti and Ae. albopictus. On the whole, permethrin resistance developed at a faster rate than malathion and temephos.
Ovitrap surveillance was conducted in two urban residential areas (Taman Samudera Timur and Taman Samudera Selatan) and in a settlement area (Kampung Banjar), which is located 16 km from Kuala Lumpur city center, Malaysia. In Taman Samudera, dengue cases were reported monthly in 2003/2004. Thus, a study was initiated to determine the distribution and abundance of dengue vectors, Aedes aegypti and Ae. albopictus. The ovitrap surveillance indicated that Ae. aegypti and Ae. albopictus were present both indoors and outdoors. The residential sites had 73 - 79% of the ovitraps with just Ae. aegypti population and Kg. Banjar had 56% of the ovitraps with just Ae. aegypti. In the indoor and outdoor of the residential areas, together with the settlement area, the Ae. aegypti density was significantly more than Ae. albopictus (p < 0.05) by 3 - 50 folds. There was no significant difference in the larval numbers of Ae. aegypti between indoors and outdoors (p > 0.05), thus implicating that adult gravid female Ae. aegypti are present both indoors and outdoors and they do oviposit indoors and outdoors. Ae. aegypti can be incriminated as the principal dengue vector in the urban residential site, Taman Samudera and in the settlement area, Kg. Banjar.