Forensically important entomological specimens recovered from 95 forensic cases of human cadavers from April 1993 to May 1996 in Malaysia were identified and analysed. The results indicated that 73.7% of these specimens were Chrysomya species, occurring either as single or mixed infestations. Of these, the most prominent species were Ch megacephala (F.) and Ch rufifacies (Macquart). Other fly maggots recovered included Sarcophaga spp., Lucilia spp. and Hermetia spp., mostly occurring together with other calliphorine flies. A member of Muscidae fly, Ophyra spp. was also recovered for the first time.
A total of 101 entomological specimens recovered from human cadavers were processed and studied. Analysis of the data indicated that about 95% of these specimens were maggots of flies. Maggots of the blowfly Chrysomya (Family: Calliphoridae) especially Ch. rufifacis and Ch. megacephala were predominantly found in 77 cases (76.2%) while larvae of several other flies of the genera Sarcophaga, Calliphora, Lucilia and hermetia were also recovered. It was notable that Musca domestica or other related flies were not found in all these specimens. The age of these larvae was useful in the determination of the minimum time lapsed after death. However, more biological studies on animal carcases should be conducted for more accurate determinations. Methods of collection, preservation and despatching of specimens were also discussed.
The ability to identify the occurrence of different resistance genotypes in field populations of mosquito is considered important for the purpose of optimising chemical control operations. The recent development of rapid microassays of enzymes responsible for resistance has provided a means for rapidly assessing the genetic background of target mosquito populations. This concept is the topic of investigation in this study. Non-specific esterase activity, which is responsible for the resistance to organophosphates in Malaysian Culex quinquefasciatus Say adults, was determined in 3 field populations from Kuala Lumpur City using rapid enzyme assay. The optical density results were used to estimate the genotypic frequencies of the populations. Subsequently, time-dependent changes in the various frequencies were determined. Such techniques allowed rapid assessment of resistance genotypes for decision-making and its possible use in insect control merits further investigation.
A screening program searching for indigenous microbial control agents of mosquitos in Malaysia is initiated since 1987 and to date at least 20 isolates of mosquitocidal Bacillus thuringiensis serotypes have been obtained. Preliminary field evaluation of several isolates indicated that they are highly effective in the control of medically important mosquito species. For operational purposes, there is an urgent need to produce this agent utilizing cheap and locally available wastes through fermentation biotechnology. Fermentation studies in shake-flasks containing standard nutrient broth and soya bean waste, respectively, indicate that it takes about 37 hours for a Malaysian isolate of B. thuringiensis serotype H-14 to mature. In the grated coconut waste, fishmeal and rice bran, the bacteria took 28 hours, 26 hours and 126 hours respectively to mature. The endotoxin was harvested from the standard nutrient broth at 55 hours and at 50 hours from soybean, grated coconut waste and fishmeal. The endotoxin could only be harvested 150 hours after inoculation from rice bran medium. However, no bacterial growth was detected in palm oil effluent. In terms of endotoxin and biomass production, fishmeal appears to be a suitable medium. Variations in the pH of the fermenting media were also noted.
Mosquitocidal strains of B. sphaericus serotype H-5a5b were shown for the first time to exhibit antagonistic activities against several human pathogens especially Salmonella. These strains of B. sphaericus also exhibited high larval toxicity against several mosquitoes.
A nationwide screening program searching for microbial control agents of mosquitos was initiated in Malaysia in 1986. A total of 725 samples were collected and 2,394 bacterial colonies were isolated and screened for larvicidal activity. From such screening, 20 Bacillus thuringiensis, 6 B. sphaericus, 1 Clostridium bifermentans and 2 Pseudomonas pseudomallei larvicidal isolates were obtained. Of these, a new B. thuringiensis named as subspecies malaysianensis was found, while the C. bifermentans was also a new anaerobe individualized as serovar malaysia. It was concluded that this screening program was highly successful.
A novel method for the control of Mansonia larvae was developed and tested. In this method, foliar absorption and translocation of a chemical insecticide, monocrotophos, a known systemic insecticide was studied in the Eicchornia plant. Acetone solution of the insecticide was painted onto leaves of the plant. At daily intervals, stems were severed and divided into equal sections which were introduced into bowls. Larvae of Aedes aegypti were tested for the presence of monocrotophos. It was found that translocation of the insecticide occurred at different rates in the stems and in some plants the chemical was also released into the surrounding water. Based on these results, 2 insecticides namely, monocrotophos and temephos were painted onto leaves of the host plant and their translocation to the root and water environment was examined by testing with Mansonia and Aedes aegypti larvae. The results again confirmed the translocation process and it was found that the insecticides were secreted into the surrounding water, thereby killing the larvae. However, in leaves painted with permethrin (synthetic pyrethroid) or flufenoxuron (chitin synthesis inhibitor), such a process was not detected. The potential of this new concept in Mansonia larval control is examined.
Comparative DDT-susceptibility status and glutathion s-transferase (GST) activity of Malaysian Anopheles maculatus, Culex quinquefasciatus and Aedes aegypti was investigated to ascertain the role of this enzyme in DDT resistance. The standardised WHO dose-mortality bioassay tests were used to determine DDT susceptibility in these mosquitos, whilst GST microassay (Brogdon and Barber, 1990) was conducted to measure the activity of this enzyme in mosquito homogenate. It appeared that DDT susceptibility status of Malaysian mosquitos was not correlated with GST activity.
The hornets are a group of venomous stinging insects that at times cause human death. A fatal case of a child stung by the lesser banded hornet Vespa affinis indosinesis is reported. Though often covered by the mass media, this constitutes the first scientifically reported case.
The bioefficacy of a commercial formulation of temephos, Creek against Aedes aegypti larvae was studied in the laboratory. Earthen jars were filled with 10 L tap water each. One g of temephos (Creek) sand granule formulation was added into each earthen jar as recommended by the manufacturer. The final test concentration of Creek was 1 mg a.i./L. One earthen jar was filled with 10 L tap water and served as a test control (untreated). Thirty late 3(rd) or early 4(th) instar of lab-bred Ae. aegypti larvae were added into each earthen jar. Mortality of the larvae was recorded after 24 hours and percent mortality was calculated. Test was repeated every week. The results showed that complete larval mortality was achieved after 24 hours. The residual effect lasted 15 weeks (105 days), indicating that Creek is effective at the dosage recommended by the manufacturer which is 1 mg a.i./L.
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.
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.
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.
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.).