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.
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 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.
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.
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.
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.
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.
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.
In this study, artificial membrane feeding technique was used to orally feed Aedes aegypti with dengue and chikungunya viruses. Virus detection was carried out by reverse transcriptase polymerase chain reaction. The study did not detect dual infection of Ae. aegypti with dengue and chikungunya virus from the same pool or from individual mosquitoes. Oral receptivity of Ae. aegypti to chikungunya virus was higher than that of dengue virus.
The importance of house fly (Musca domestica L) wings in mechanical transmission of bacteria was studied. A droplet of phosphate-buffered saline containing Vibrio cholerae was rolled along one wing of each house fly. None adhered to the wings but small proportions of the bacterium were isolated from about half the wings. Vibrio cholerae was spread onto the ventral wing surfaces of each unconscious house fly which then was placed inside a bottle. When it regained consciousness, the types of activity it performed over five minutes were noted before the house fly was killed and the bacteria on its wings numerated. Control were house flies killed before inoculation. The proportion of house flies with bacteria on their wings and the mean number of bacteria remaining were significantly less on live house flies than killed controls. Among the live house flies, bacteria were detected on fewer house flies which flew (25%) than those which did not fly (81%). In addition, the mean number of bacteria on the former was significantly less than the latter (5 against 780 colonies). However, both these parameters were not significantly different between the group which performed and the group which did not perform wing grooming; takeoff and alighting over short distances, and somersaulting. Wings of unconscious house flies tethered by their thoraxes were inoculated with V. cholerae. After regaining consciousness, the house flies were allowed to move their wings in flight motions for up to 30 seconds. Small proportions of bacteria remained on all the house flies. House flies were placed in a chamber containing a liquid bait spiked with V. cholerae. After two hours, 10 were removed sequentially and cultured for V. cholerae. The bacterium was isolated from four house flies: two from the legs, and two others from their bodies minus legs and wings. In conclusion, house fly wings do not play an important role in mechanical transmission of bacteria suspended in a non-adhering liquid medium because of the low transfer rate of the bacteria to the wings and poor retention of bacteria on the wings during normal house fly activities.
A study was conducted to examine the persistency of transovarial dengue virus type 2 (DEN-2) in a Selangor strain of Aedes aegypti mosquitoes. Two hundred 4-5 day old female mosquitoes were fed with blood containing dengue virus. The infected mosquitoes were reared to the 7th generation; each generation was screened for the virus using immunological staining methods. The virus was detectable until the 5th generation but absent in the 6th and the 7th generations. Therefore, dengue virus type 2 can be transmitted transovarially in Aedes aegypti mosquitoes until the fifth generation under laboratory conditions.
A study was carried out to investigate correlation between presence of specific microalgal species and Leptoconops biting midge larvae in its breeding habitats. Sand samples containing microalgae were collected from the beach where the midges were most commonly biting and from sand beaches which are potential as breeding habitats of Leptoconops. The survey covered sand beaches from 12 seperate islands. At all sites, the Bacillariophyta constituted the largest representatives of the microalgae community with the majority from the Naviculaceae family. A total of 24 microalgal species were identified from the sand samples collected from the study sites. Sand samples from Kentot Kecil Island had the highest number of algal species (11.0) and the highest algae species diversity ( Shanon-Weiner Diversity Index, H' = 0.884). Besar Island (Johor) had the lowest number of algal species (2.0) whereas Tengol A Island had the lowest algae species diversity (H'=0.234). Highest similarity index was recorded between sand samples collected from Tengol A Island and Tengol B Island (75.0%) followed by Besar Island (Melaka) and Tengol B Island (62.0 %). The variation between other islands were relatively high. Virtually many kinds of algae were found where Leptoconops were breeding but Fragilaria intermedia, Mastigloia minuta and Navicula advena were particularly common.