The present study explored the intraspecific genetic diversity, dispersal patterns and phylogeographic relationships of Culex quinquefasciatus Say (Diptera: Culicidae) in Malaysia using reference data available in GenBank in order to reveal this species' phylogenetic relationships. A statistical parsimony network of 70 taxa aligned as 624 characters of the cytochrome c oxidase subunit I (COI) gene and 685 characters of the cytochrome c oxidase subunit II (COII) gene revealed three haplotypes (A1-A3) and four haplotypes (B1-B4), respectively. The concatenated sequences of both COI and COII genes with a total of 1309 characters revealed seven haplotypes (AB1-AB7). Analysis using tcs indicated that haplotype AB1 was the common ancestor and the most widespread haplotype in Malaysia. The genetic distance based on concatenated sequences of both COI and COII genes ranged from 0.00076 to 0.00229. Sequence alignment of Cx. quinquefasciatus from Malaysia and other countries revealed four haplotypes (AA1-AA4) by the COI gene and nine haplotypes (BB1-BB9) by the COII gene. Phylogenetic analyses demonstrated that Malaysian Cx. quinquefasciatus share the same genetic lineage as East African and Asian Cx. quinquefasciatus. This study has inferred the genetic lineages, dispersal patterns and hypothetical ancestral genotypes of Cx. quinquefasciatus.
Monodispersed iron oxide nanoparticles (IONPs) coated with polystyrenesulfonate (PSS) and cetrimonium bromide (CTAB) have been used to stabilize magnetic Pickering emulsions (MPEs). Magnetophoresis of MPEs under the influence of a low gradient magnetic field (∇B < 100 T/m) was investigated at the macroscopic and microscopic scale. At the macroscopic scale, for the case of pH 7, the MPE achieved a magnetophoretic velocity of 70.9 μm/s under the influence of ∇B at 93.8 T/m. The magnetic separation efficiency of the MPE at 90% was achieved within 30 min for pH 3, 7, and 10. At pH 10, the colloidal stability of the MPE was the lowest compared to that for pH 3 and 7. Thus, MPE at pH 10 required the shortest time for achieving the highest separation efficiency, as the MPE experienced cooperative magnetophoresis at alkaline pH. The creaming rate of the MPE at all conditions was still lower compared to magnetophoresis and was negligible in influencing its separation kinetics profiles. At the microscopic scale, the migration pathways of the MPEs (with diameters between 2.5 and 7.5 μm) undergoing magnetophoresis at ∇B ∼ 13.0 T/m were recorded by an optical microscope. From these experiments, and taking into consideration the MPE size distribution from the dynamic light scattering (DLS) measurement, we determined the averaged microscopic magnetophoretic velocity to be 7.8 ± 5.5 μm/s. By making noncooperative magnetophoresis assumptions (with negligible interactions between the MPEs along their migration pathways), the calculated velocity of individual MPEs was 9.8 μm/s. Such a value was within the percentage error of the experimental result of 7.8 ± 5.5 μm/s. This finding allows for an easy and quick estimation of the magnetophoretic velocity of MPEs at the microscale by using macroscopic separation kinetics data.
The compatibility of the commercial aqueous Bacillus thuringiensis serovar israelensis (B.t.i.) formulation, Vectobac 12AS, with the chemical insecticides Actellic 50EC, Aqua Resigen, Resigen, and Fendona SC, for the simultaneous control of Aedes larvae and adults was studied by dispersing nine different formulations using a portable mist blower, in single story half-brick houses. The effectiveness of the treatment was evaluated by measuring the larval mortality, adult mortality, and droplet analysis at varying distances from the sprayer. Persistence of the larvicidal activity of the chemical insecticides and B.t.i was also determined by measuring the larval mortality in the test samples 7 days posttreatment. The sprayed particles in all the trials were 50-60 microns in size, indicating that the particles were those of mist spray. Test samples placed within 3 m from the sprayer gave the maximum larval and adult mortality. Chemical insecticides exhibited maximum larval mortality in the 1 h posttreatment test samples and it was comparable to the larvicidal activity of B.t.i. The larvicidal toxins of B.t.i were more stable and were able to affect sufficient larval mortality for 7 days posttreatment. The larvicidal activity of the mixtures, i.e., chemical insecticides with B.t.i, in the 1 h posttreatment test samples was not significantly different from the larvicidal activity of the chemical insecticides and it was comparable to the larvicidal activity of B.t.i alone. However, the larvicidal activity of the mixtures was significantly more than the chemical insecticides alone in the 7 days posttreatment test samples except for the Actellic 50EC and Vectobac 12AS mixture. In all the trials, with or without B.t.i, there was no significant difference in adult mortality, indicating that this B.t.i formulation, Vectobac 12AS, was not antagonistic to the adulticidal activity of the chemical insecticides. From this study, it can be concluded that chemical insecticides can be used effectively for both adult and larval control, but the chemical insecticides do not exhibit residual larvicidal activity. Hence, for an effective control of both Aedes larvae and adults, it is advisable to add B.t.i. to the chemical insecticides, as B.t.i is specifically larvicidal and is also able to effect extended residual larvicidal activity.
Field strains of house flies, Musca domestica L., from three different breeding sites-garbage dump (IMR), poultry farm (Kundang, Selangor), and agricultural farm (Kampung Batu, Kuala Lumpur), were evaluated against five insecticides. Resistance status of adult female flies was determined using the modified WHO bioassay methods. The WHO susceptibility strain was used as a reference strain for comparison. Flies from the garbage dump and poultry farm were more resistant to the insecticides than the strain from the agricultural farm. Results obtained from bioassay tests were confirmed by in-vitro microenzyme assays of non-specific esterases and glutathione-S-transferases. Significant differences between the esterase levels of WHO and field strains were observed. Levels of glutathion-S-transferases were approximately the same, which may indicate that other enzymes are involved in house fly resistance.
The combined adulticidal, larvicidal, and wall residual activity of ULV-applied bifenthrin, a synthetic pyrethroid, was evaluated in houses in Kuala Lumpur, Malaysia, against larvae and adults of lab-bred Aedes aegypti and Culex quinquefasciatus. A portable ULV sprayer was used to disperse a ULV formulation of bifenthrin at a discharge rate of 45 ml/min. The results indicated that bifenthrin sprayed at this rate exhibited all the three activities against the test mosquitoes. Complete adult mortalities were achieved, while very high larvicidal activity was also effected, which persisted for seven days. Wall bioassay with adults of Ae aegypti also resulted in very high mortality, which also persisted for one week. The combined mosquitocidal activities of bifenthrin is considered more effective especially in the control of dengue vectors.
BACKGROUND & OBJECTIVESI: Transovarial transmission of dengue virus in the Aedes vectors is now a well-documented phenomenon reported from many parts of the endemic areas in the world, which played an important role in initiating and maintaining the outbreak in human populations. This study investigated the factors affecting breeding habitats and the relationship with transovarial dengue virus in larvae of Aedes aegypti and Ae. albopictus.
A novel Clostridium bifermentans strain toxic to mosquito larvae on ingestion was isolated from a soil sample collected from secondary forest floor. This strain was designated as serovar paraiba (C. b. paraiba) according to its specific H antigen. Clostridium bifermentans paraiba is most toxic to Anopheles maculatus Theobald larvae (LC50 = 0.038 mg/liter), whereas toxicity to Aedes aegypti (Linn.) (LC50 = 0.74 mg/liter) and Culex quinquefasciatus Say (LC50 = 0.11 mg/liter) larvae was 20 and 3 times lower, respectively. The toxicity to An. maculatus larvae is as high as that of Bacillus thuringiensis serovar israelensis. C. b. paraiba was also found to exhibit significant per os insecticidal activity toward adult Musca domestica (Linn.).
Evaluation of the effectiveness of Bacillus thuringiensis ssp. israelensis (B.t.i.) against mosquito larvae dispersed by ultralow volume (ULV) spraying was conducted in simulated field trials. Effectiveness was measured using 3 different indicators: larval mortality, colony-forming unit enumeration, and droplet analysis. B.t.i. was dispersed with a ULV generator using 2 different flow rates: 0.3 and 0.5 liter/min on 2 different days. Based on the results of this study, it can be concluded that an output of 0.3 liter/min is effective for controlling Aedes aegypti. although a dosage of 0.5 liter/min can be used when high residual activity is desired. For Culex quinquefasciatus control, both dosages were effective but with low residual activity. For Anopheles maculatus control, only a discharge rate of 0.5 liter/min was effective with low residual activity. B.t.i. application at both dosages penetrated tires well, indicating that B.t.i. ULV application is an effective method for controlling container-inhabiting mosquitoes. Good coverage of target area and penetration were attributed to satisfactory droplet profiles.
A novel Bacillus thuringiensis strain highly toxic to mosquitoes was isolated from soil samples in Malaysia. This strain was shown to display a new subfraction of the H-28 flagellar antigen determining a new serovar H28a28c, which was designated serovar jegathesan. Bioassays indicated that Culex quinquefasciatus larvae are the most susceptible to this new isolate, whereas toxicity to Anopheles maculatus and Aedes aegypti larvae was 10 times lower. The potency of this new serotype is also comparable to most of the Malaysian B. thuringiensis H-14 isolates.
The present study aims to investigate the susceptibility status of Aedes albopictus, Culex quinquefasciatus, and Cx. vishnui collected from a pig farm in Tanjung Sepat, Selangor, toward 11 insecticides representing the classes of organochlorines, carbamates, organophosphates, and pyrethroids. The results of a World Health Organization adult mosquito bioassay revealed that Ae. albopictus, Cx. quinquefasciatus, and Cx. vishnui exhibited different susceptibility toward various insecticides. Overall, pyrethroids were able to induce rapid knockdown for all test mosquito species. The pyrethroids lambdacyhalothrin and etofenprox were able to cause high mortality (> 80%) of all 3 species. The findings of the present study will benefit local authorities in selecting appropriate dosage of insecticides to be used in mosquito control in this area.
A standardized larval dipping method was used to determine the infestation rates of Culex and other species of mosquitoes in stagnant water at 20 residential areas. This study also examined the associations between Culex distribution and various habitat characteristics across all states in Malaysia. Identification of 7,848 specimens yielded 6 species dominated by Culex quinquefasciatus (82.74%), followed by Cx. vishui (14.39%), Cx. gelidus (2.70%), Lutzia fuscanus (0.11%), Armigeres subalbatus (0.05%), and Anopheles separatus (0.01%). The Culex larvae occurred in stagnant water with pH ranging from 6.4 to 8.2; conductivity, 139.7 to 6635.2 micros/cm; salinity, 0.07 to 3.64 ppt; total dissolved solids, 0.09 to 4.27g/liter; and dissolved oxygen, 5.11 to 8.11 mg/liter. The mean number of Culex larvae was positively correlated with pH, conductivity, salinity, and total dissolved solids. In contrast, the elevation and dissolved oxygen were found negatively correlated with mean number of Culex larvae. This study documented baseline information on the habitat characteristics of Culex species for the 1st time at different residential areas in Malaysia. The findings of this study will be a timely reminder to local authorities that effective control measures should be monitored regularly in order to reduce the nuisance of these mosquitoes and the risks of disease transmission.
A nationwide investigation was carried out to determine the current susceptibility status of Culex quinquefasciatus Say populations against four active ingredients representing four major insecticide classes: DDT, propoxur, malathion, and permethrin. Across 14 study sites, both larval and adult bioassays exhibited dissimilar trends in susceptibility. A correlation between propoxur and malathion resistance and between propoxur and permethrin resistance in larval bioassays was found. The results obtained from this study provide baseline information for vector control programs conducted by local authorities. The susceptibility status of this mosquito should be monitored from time to time to ensure the effectiveness of current vector control operations in Malaysia.
Susceptibility status of Aedes aegypti (L.) and Aedes albopictus Skuse larvae obtained from 12 states in Malaysia were evaluated against five insect growth regulators (IGRs), namely, pyriproxyfen, methoprene, diflubenzuron, cyromazine, and novaluron under laboratory conditions. Field populations of Ae. aegypti exhibited moderate resistance toward methoprene and low resistance toward pyriproxyfen, with resistance ratios of 12.7 and 1.4, respectively, but susceptibility to diflubenzuron, cyromazine, and novaluron. On the other hand, field populations of Ae. albopictus exhibited low resistance against diflubenzuron and novaluron, with resistance ratio of 2.1 and 1.0, respectively, but susceptibility to other tested IGRs. Our study concluded that the tested IGRs provide promising results and can be used to control field population of Ae. aegypti and Ae. albopictus, especially cyromazine. The use of IGR should be considered as an alternative when larvae develop resistance to conventional insecticides.
Factors affecting the mechanical transmission of rotavirus by the legs and wings of the housefly, Musca domestica L., were examined in a laboratory study. Rotavirus was picked up when houseflies walked on thin smears of clarified rotavirus suspensions. The addition of glycerol, which increased viscosity of the virus suspension, and particulate human feces slightly increased the proportion of flies contaminated with virus. However, the addition of glycerol greatly reduced the average number of virus particles picked up per fly, whereas feces greatly increased the number of particles. The proportion of flies with virus-contaminated legs, which transferred virus to > 1 contact surface, was increased by longer contact time with the surface and when the contact surface was agar instead of glass. Most virus particles were deposited on 1st contact with the surface. Most flies dislodged virus particles inoculated on the underside of their wings soon after the start of simulated flight. Our data indicated that the nature of the virus-suspending medium has a greater effect on the level of virus contamination than on the ability to become contaminated. The importance of walking as a mode of virus transport depends on the nature of the contact surface, the risk of the contaminated fly settling first on a surface likely to come into contact with humans, and fly numbers.
Resistance status of Aedes albopictus (Diptera: Culicidae) collected from Sabah, East Malaysia, was evaluated against four major classes of adulticides, namely pyrethroid, carbamate, organochlorine, and organophosphate. Adult bioassays conforming to WHO standard protocols were conducted to assess knockdown and mortality rates of Ae. albopictus. Among tested pyrethroid adulticides, only cyfluthrin, lambda-cyaholthrin, and deltamethrin were able to inflict total knockdown. The other adulticide classes mostly failed to cause any knockdown; the highest knockdown rate was only 18.33% for propoxur. With regards to mortality rate, Ae. albopictus was unanimously susceptible toward all pyrethroids, dieldrin, and malathion, but exhibited resistance toward bendiocarb, propoxur, dichlorodiphenyltrichloroethane, and fenitrothion. Additionally, correlation analysis demonstrated cross-resistance between bendiocarb and propoxur, and malathion and propoxur. In conclusion, this study has disclosed that pyrethroids are still generally effective for Aedes control in Sabah, Malaysia. The susceptibility status of Ae. albopictus against pyrethroids in descending order was cyfluthrin > lambda-cyhalothrin > deltamethrin > etofenprox > permethrin.
The bioefficacy of commercial mosquito coils containing four different active ingredients, namely metofluthrin, d-allethrin, d-trans allethrin, and prallethrin against Aedes albopictus (Skuse) (Diptera: Culicidae) from 10 states in Malaysia, was evaluated using the glass chamber method. In this study, Ae. albopictus exhibited various knockdown rates (50% knockdown time, KT50), ranging from 2.50 to 5.00 min, 2.50 to 7.00 min, 3.00 to 8.00 min, and 5.00 to 17.00 min for metofluthrin, d-trans allethrin, d-allethrin, and prallethrin, respectively. Overall, all strains of Ae. albopictus were most susceptible to metofluthrin, with mortality rates >80%. On the other hand, mortality rates ranging from 5.0 to 100% were observed from all populations exposed to d-trans allethrin, d-allethrin, and prallethrin. In addition, significant correlations between KT50 of metofluthrin and d-allethrin (r = 0.758, P = 0.011), metofluthrin and prallethrin (r = 0.676, P = 0.032), d-allethrin and d-trans allethrin (r = 0.832, P = 0.003), d-allethrin and prallethrin (r = 0.921, P = 0.000), and d-trans allethrin with prallethrin (r = 0.941, P = 0.000) were detected, suggesting some levels of cross-resistance within the pyrethroid insecticides. This study demonstrated that metofluthrin can induce high insecticidal activity in Ae. albopictus in Malaysia, followed by d-trans allethrin, d-allethrin, and prallethrin.
Resistance status of Aedes albopictus (Skuse) from 13 districts in Sarawak State, Malaysia, was evaluated against four major classes of adulticides, namely organochlorine, organophosphate, carbamate, and pyrethroid. Adult bioassays were performed according to the World Health Organization (WHO) standard protocols to assess knockdown and mortality rates of Ae. albopictus. Among the tested pyrethroids, only cyfluthrin was able to exhibit complete knockdown. On the other hand, different susceptibility and resistance patterns were observed in other adulticides. As for mortality rates, the mosquitoes were susceptible to cyfluthrin and dieldrin but exhibited various susceptibilities to other tested adulticides. Cross-resistance was discovered within and between tested insecticide classes. Significant correlations were found within pyrethroid and carbamate classes (i.e., bendiocab and propoxur, P = 0.036; etofenprox and permethrin, P = 0.000; deltamethrin and lambda-cyhalothrin, P = 0.822; deltamethrin and permethrin, P = 0.042). Additionally, insecticides belonging to different groups were also found significantly correlated (i.e., malathion and deltamethin, P = 0.019; malathion and bendiocarb, P = 0.008; malathion and propoxur, P = 0.007; and bendiocarb and deltamethrin, P = 0.031). In conclusion, cyfluthrin was effective for Aedes albopictus control in Sarawak State and these data may assist local authorities to improve future vector control operations.
The efficacy of three groups of insect growth regulators, namely juvenile hormone mimics (methoprene and pyriproxyfen), chitin synthesis inhibitors (diflubenzuron and novaluron), and molting disruptor (cyromazine) was evaluated for the first time, against Aedes albopictus Skuse (Diptera: Culicidae) larvae from 14 districts in Sabah, Malaysia. The results showed that all field populations of Ae. albopictus were susceptible towards methoprene, pyriproxyfen, diflubenzuron, novaluron, and cyromazine, with resistance ratio values ranging from 0.50-0.90, 0.60-1.00, 0.67-1.17, 0.71-1.29, and 0.74-1.07, respectively. Overall, the efficacy assessment of insect growth regulators in this study showed promising outcomes and they could be further explored as an alternative to conventional insecticides.
Forensic entomological specimens collected from human decedents during crime scene investigations in Malaysia in the past 6 years (2005-2010) are reviewed. A total of 80 cases were recorded and 93 specimens were collected. From these specimens, 10 species of cyclorrphagic flies were identified, consisting of Chrysomya rufifacies (Macquart) -38 specimens (40.86%), Chrysomya megacephala (Fabricius) -36 specimens (38.70%), Chrysomya villeneuvi (Patton) -2 specimens (2.15%), Chrysomya nigripes (Aubertin) -2 specimens (2.15%), Chrysomya pinguis (Walker) -1 specimen (1.08%), Hermetia illucens (Linnaeus) -1 specimen (1.08%), Hemipyrellia liguriens (Wiedemann) -5 specimens (5.37%), Synthesiomyia nudiseta (Wulp) -1 specimen (1.08%), Megaselia scalaris (Loew)-1 specimen (1.08%) and Sarcophaga ruficornis (Fabricius) -4 specimens (4.30%). In two specimens (2.15%), the maggots were not identifiable. Ch. megacephala and Ch. rufifacies were the commonest species found in human decedents from three different ecological habitats. S. nudiseta is an uncommon species found only on human cadavers from indoors. A total of 75 cases (93.75%) had a single fly infestation and 5 cases (6.25%) had double fly infestation. In conclusion, although large numbers of fly species were found on human decedents, the predominant species are still those of Chrysomya.