Isolation of Japanese encephalitis virus (JEV) from mosquitoes in Sabak Bernam, Selangor, Malaysia, was attempted. An aliquot of homogenate from each pool of mosquitoes, 50 per tube, was inoculated into Aedes albopictus clone C6/36 cells for virus isolation. Each cell culture was tested for the presence of viral antigen by immunoperoxidase staining using an anti-JEV polyclonal antibody. Out of 4 Culex sitiens mosquito pools, 2 pools were positive for JEV by cell culture. Presence of JEV genome in the cell cultures for Cx. sitiens was confirmed by using reverse transcriptase-polymerase chain reaction and JEV-specific primers. This is the first report on the isolation of JEV from Cx. sitiens.
Systematic surveys of the wild macaques of South Asia by blood culture resulted in the discovery that trypanosomiasis is enzootic in the simians of Indonesia, Malaysia, India, and Thailand. The isolates obtained differ in morphology, metabolism, and ability to multiply in arthropods. Following this discovery, interest focused on possible transmissions of these trypanosomiases. Laboratory-reared and wild-caught insects were studied to determine which are satisfactory intermediate hosts and potential natural vectors. Successful results were obtained with insectary-reared reduviids and Indonesian isolates. In Rhodnius prolixus and Triatoma rubrofasciata the Indonesian trypanosomes multiply for periods which can exceed 40 days. The flagellate infections are in the digestive tract, whereas trypanosomes have never been seen in the salivary glands or in the hemolymph. The feces of trypanosome-carrying reduviids are infective, suggesting a stercoreal method of infection of mammals, and infection was produced in experiments in which feeding by the insects was not possible. The relevance of these findings to natural transmission in Indonesia is not known. Experiments with insects and all other trypanosomal isolates have been negative. The natural transmission mechanism(s) of the simian trypanosomiases in South Asia remains an unsolved problem.
The mosquito Anopheles balabacensis balabacensis has been identified as a natural vector of at least two species of simian malaria in the monsoon forests of the northern Malay States. This mosquito is also a serious vector of human malaria from Viet Nam to northern Malaya. This is the first report of a mosquito which transmits both human and simian malaria in nature.
Mosquito collections were carried out for a period of one year from January to December 1992 in a pig farm in Sungai Pelek, Selangor, Malaysia. A total of 41,022 mosquitos belonging to 52 species and 20 genera were collected. Culex tritaeniorhynchus and Cx. gelidus, the important vectors, comprised 63% of all mosquitos collected. Both these species were collected in large numbers during the wet months of May and December. The other predominant species in that area were Cx. fuscocephala, Cx. quinquefasciatus, Cx. sitiens, Aedes butleri, and Armigeres subalbatus.
Five mark-release-recapture experiments with wild caught Ma. uniformis were conducted in an open swamp area at Batang Berjuntai in Selangor, 40 km from Kuala Lumpur, Malaysia, between May 1983 and January 1985. A total of 64 (0.14%) from the 45,950 females released were recaptured feeding on humans and cattle and resting in cattle-sheds. Substantially fewer (0.03% to 0.09%) females were recaptured from releases of blood-fed females than from releases of unfed females (0.20% to 0.23%). More than 70% of all recaptures were taken within a radius of 1.5 km around the point of release and the longest detected flight was 3.5 km. The mean dispersal distance for blood-fed and unfed females was 1.445 +/- 1.06 and 1.706 +/- 1.03 km, respectively. However, there was no significant difference in the overall mean dispersal of the two groups of females (p greater than 0.05). The duration of the gonotrophic cycle in the field was between 3 to 4 days. Daily survivorship estimates (0.783-0.867) based on the recapture rates of date specific marked females was comparable to that estimated vertically from the dissection of unmarked females (0.751-0.795). These experiments revealed the remarkable flying ability of Ma. uniformis and the importance of reinvasion must be recognized when control operations are restricted to small areas.
A total of 37 species of mosquitoes from seven genera were collected in six villages in the Bengkoka Peninsula, Sabah State, during two visits in 1981 in connection with studies on malaria and filariasis. Fifty-five per cent of the total mosquitoes collected were Mansonia. An. collessi constituted a new record of the species from Sabah. An. balabacensis was found to be naturally infected with sporozoites. Ma. bonneae was found to be naturally infected with Brugia, probably B. malayi. Parous rates of An. balabacensis and Ma. bonneae were very high with consequent high probability of survival ideally suiting transmission of malaria and filariasis.
Comparative studies of vector efficiency were done with the Liverpool and Malaysian strains of Aedes (Finlaya) togoi for subperiodic Brugia malayi and Brugia pahangi. The Malaysian strain of A. togoi was found to take in fewer microfilariae under the same experimental conditions than the Liverpool strain. Also, for various microfilarial densities in the host's peripheral blood, the Malaysian strain had less mean infective larvae per fed mosquito than the Liverpool strain. The microfilarial intake of A. togoi was not affected by the site of feeding on the host affected by the site of feeding on the host. Most of the mosquitoes took in fewer microfilariae than expected. It is concluded from these studies that the Malaysian strain of A. togoi is a susceptible and reasonably good vector for subperiodic B. malayi and B. pahangi. Further field studies should be carried out to determine its importance as a natural vector of Brugian filariasis.
An experiment was carried out with Mansonia mosquitos in an area endemic for subperiodic Brugia malayi to assess the applicability of the mark-release-recapture method to these mosquitos. An estimated 17,880 individuals of six species of Mansonia were marked with fluorescent dust and released: 453 Ma. annulata, 305 Ma. annulifera, 6,200 Ma. bonneae, 516 Ma. dives, 3,998 Ma. indiana and 6,408 Ma. uniformis. Twenty-three marked individuals were recaptured. Most recaptures were made one or two nights after their release, but one Ma. annulifera was recaptured five nights later and one Ma. bonneae had been marked and released 6-11 nights previously. The recaptured mosquitos were collected between 0.5 and 2.4 km from their release points.
Detection and isolation of Japanese encephalitis (JE) virus were attempted from female mosquitoes collected in Kampong Pasir Panjang, Sabak Bernam, Selangor, from May to November 1992. A total of 7,400 mosquitoes consisting of 12 species in 148 pools were processed and inoculated into Aedes albopictus clone C6/36 cell cultures. Of these, 26 pools showed the presence of viral antigens in the infected C6/36 cells by specific immunoperoxidase staining using an anti-JE virus polyclonal antibody. Presence of JE virus genome was confirmed in the infected culture fluid for 16 pools by using reverse transcriptase-polymerase chain reaction and JE virus-specific primers. Of these, 3 pools were from Culex tritaeniorhynchus, 4 from Culex vishnui, 3 from Culex bitaeniorhynchus, 2 from Culex sitiens, one from Aedes species, and 3 from Culex species. Isolation of JE virus from Cx. sitiens, Cx. bitaeniorhynchus, and Aedes sp. (Aedes butleri and Ae. albopictus) is reported for the first time in Malaysia.