Malaria and filariasis still continue to pose public health problems in developing countries of the tropics. Although plans are in progress for the elimination of both these parasitic vector borne diseases, we are now faced with a daunting challenge as we have a fifth species, Plasmodium knowlesi a simian malaria parasite affecting humans. Similarly in peninsular Malaysia, filariasis was mainly due to Brugia malayi. However, we now see cases of Wuchereria bancrofti in immigrant workers coming into the country. In order to successfully eliminate both these diseases we need to know the vectors involved and introduce appropriate control measures to prevent the diseases occurring in the future. As for knowlesi malaria it is still uncertain if human to human transmission through mosquito bites is occurring. However, P. knowlesi in human is not a rare occurrence anymore and has all the characteristics of a pathogen spreading due to changes in the ecosystem, international travel, and cross border migration. This has created a more complex situation. In order to overcome these challenges we need to revamp our control measures. This paper reviews the vectors of malaria and filariasis in Southeast Asia with special emphasis on P. knowlesi and W. bancrofti in Malaysia and their control strategies.
Plasmodium knowlesi in humans is life threatening, is on the increase and has been reported from most states in Malaysia. Anopheles latens and Anopheles cracens have been incriminated as vectors. Malaria is now a zoonoses and is occurring in malaria free areas of Malaysia. It is also a threat to eco-tourism. The importance of the vectors and possible control measures is reviewed here.
Five pyrethroids namely, lambda-cyhalothrin, permethrin, deltamethrin, cyfluthrin and alpha-cypermethrin were evaluated using adult male and female cockroaches, Periplaneta americana (L). The American cockroaches were exposed for 10 minutes to glass jars treated with different concentrations of the five pyrethroids. The cockroaches were susceptible to all five pyrethroids and the susceptibility based on LC50 and LC50 were observed to be in the following order: lambda-cyhalothrin > permethrin > deltamethrin > cyfluthrin > alpha-cypermethrin The results showed that lambda-cyhalothrin was the most effective and alpha-cypermethrin was the least effective against Periplaneta americana (L).
Field trials were conducted in two residential areas of Petaling Jaya Municipality to test the adulticidal and larvicidal effects of malathion 96% TG and Resigen on Aedes aegypti. Malathion is the currently used insecticide in Malaysia for the control of dengue. The Leco HD ULV machine was used throught the trials. For malathion the flow rate was 90 ml/minute at a vehicle speed of 8kph and for Resigen the flow rate was 200 ml/minute at the same vechicle speed. Malathion was more effective giving higher mortality rates when compared with Resigen. The mortality rate of adult Ae. aegypti outdoor was higher than in the living room and kitchen. Both insecticides did not show promising larvicidal effects.
Plasmodium knowlesi is a malaria parasite of Old World monkeys and is infectious to humans. In this study Macaca fascicularis was used as a model to understand the host response to P. knowlesi using parasitological and haematological parameters. Three M. fascicularis of either sex were experimentally infected with P. knowlesi erythrocytic parasites from humans. The pre-patent period for P. knowlesi infection in M. fascicularis ranged from seven to 14 days. The parasitemia observed was 13,686-24,202 parasites per microL of blood for asexual stage and 88-264 parasites per microL of blood for sexual stage. Periodicity analysis adopted from microfilaria periodicity technique of asexual stage showed that the parasitemia peak at 17:39h while the sexual stage peaked at 02:36 h. Mathematical analysis of the data indicates that P. knowlesi gametocytes tend to display periodicity with a peak (24:00-06:00) that coincides with the peak biting activity (19:00-06:00) of the local vector, Anopheles latens. The morphology of P. knowlesi resembled P. falciparum in early trophozoite and P. malariae in late trophozoite. However, it may be distinguishable by observing the appliqué appearance of the cytoplasm and the chromatin lying inside the ring. Haematological analysis on macaques with knowlesi malaria showed clinical manifestations of hypoglycaemia, anaemia and hyperbilirubinemia. Gross examination of spleen and liver showed malaria pigments deposition in both organs.
A preliminary survey of Aedes aegypti was carried out in 6 areas in the state of Selangor, Malaysia. The densities of Ae. aegypti and Ae. albopictus in the areas were discussed. Results indicated that the distribution of larval habitats varied with the housing type. The most ommon indoor larval habitat in urban areas is the bathroom tank. In both urban and rural areas, outdoor preference is for the earthenware jars. Anttraps have decreased in importance as larvae breeding habitats. The accuracy of house searches can be increased by increasing the number of larvae examined per container to 5 or more. Further study is required to determine whether the findings of this survey is peculiar to the areas surveyed or is representative of the whole country, and whether there is a seasonal fluctuation in the types of preferred larval habitats.
Accurate identification of filarial parasites in mosquitoes poses a major problem for the coordination of filariasis control programs. Traditional methods are tedious, and some are not specific enough to give satisfactory results. Amplification of specific gene sequences by primer-directed polymerase chain reaction (PCR) has been increasingly utilized as a diagnostic tool. However, current protocols for the extraction of parasite DNA from mosquito samples are tedious and could lead to failure of PCR amplification. We demonstrate that the use of Chelex is an efficient method for DNA extraction from mosquitoes and the parasite and that PCR amplification with primers specific for Brugia malayi yields a band of the expected size. The PCR products were transferred to a nylon membrane with Southern blotting, and a B. malayi-specific digoxigenin-labeled probe confirmed the sequence similarity of the PCR-amplified fragment and increased the sensitivity of the PCR assay. Use of this probe enabled us to detect PCR-amplified product from B. malayi even when a product was not visible on an ethidium bromide-stained agarose gel. This increased sensitivity allowed us to detect the parasite in the heads of mosquitoes.
Adult mosquito collections were conducted for 12 weeks in two residential areas in Kuala Lumpur. The CDC light traps were compared using dry ice and yeast as sources of carbon dioxide attractants for mosquitoes. The efficacy of the dry ice baited trap was significant over yeast generated CO2 trap. The predominant species obtained were Culex quinquefasciatus, Stegomyia albopicta and Armigeres subalbatus.
Malaria continues to be a global public health problem although it has been eliminated from many countries. Sri Lanka and China are two countries that recently achieved malaria elimination status, and many countries in Southeast Asia are currently in the pipeline for achieving the same goal by 2030. However, Plasmodium knowlesi, a non-human primate malaria parasite continues to pose a threat to public health in this region, infecting many humans in all countries in Southeast Asia except for Timor-Leste. Besides, other non-human primate malaria parasite such as Plasmodium cynomolgi and Plasmodium inui are infecting humans in the region. The non-human primates, the long-tailed and pig-tailed macaques which harbour these parasites are now increasingly prevalent in farms and forest fringes close by to the villages. Additionally, the Anopheles mosquitoes belonging to the Lecuosphyrus Group are also present in these areas which makes them ideal for transmitting the non-human primate malaria parasites. With changing landscape and deforestation, non-human primate malaria parasites will affect more humans in the coming years with the elimination of human malaria. Perhaps due to loss of immunity, more humans will be infected as currently being demonstrated in Malaysia. Thus, control measures need to be instituted rapidly to achieve the malaria elimination status by 2030. However, the zoonotic origin of the parasite and the changes of the vectors behaviour to early biting seems to be the stumbling block to the malaria elimination efforts in this region. In this review, we discuss the challenges faced in malaria elimination due to deforestation and the serious threat posed by non-human primate malaria parasites.
Malaria vector surveys were carried out in 8 provinces in Lao PDR in 1999. The surveys were conducted in 4 provinces - Savannakhet, Champasak, Luang Perbang and Sayaboury in May and in another 4 provinces - Bolikhamsay, Sarvan, Sekong and Vientiane in December 1999. Bare leg collection were carried out indoors and outdoors from 6 pm to 5 am. All anopheline mosquitos were identified, dissected and the gut, gland and ovaries were examined. A total of 438 Anopheles mosquitos belonging to 19 species were obtained. Of these only 3 species were found to be infected with oocysts - An. maculatus, An. dirus and An. minimus. All these species were found biting both indoors and outdoors. An. aconitus was the predominant species obtained in the December collection but its vectorial status remains unknown.
Two formulations of lambda-cyhalothrin (EC-Emulsion concentrate and MC-Microencapsulated) were impregnated into bednets made of polyethylene and polyester. The nets were treated at a dosage of 15 mg/m2. For bioassay of insecticidal efficacy, female Anopheles maculatus and Aedes aegypti were exposed to the nets for two minutes and mortality was scored 24 hours later. The nets were also tested after repeated washings with water and with soap and water. Microencapsulated (2.5CS) formulation was more effective than emulsion concentrate (2.5EC) formulation on both net materials--polyethylene and polyester. Repeated washing with water and soap reduces the efficacy of all bednet treatment combinations. Microencapsulated formulation on polyethylene gave best results; it could sustain up to five washes with water and two with soap and water.
The efficacy of two formulations, wettable powder and emulsifiable concentrate, of cyfluthrin sprayed on plywood [10 mg (ai)/m2] was assessed against six species of mosquitos. The bioassay followed the WHO standard method, with some modification for the bioassay of insecticidal deposits on wall surfaces. The results indicated that these two formulations of cyfluthrin were effective against Anopheles dirus and Mansonia uniformis, moderately toxic to Aedes aegypti and Ae. albopictus in decreasing mortality through out the study period. It was least effective against Culex quinquefasciatus and An. maculatus, respectively. There was no statistically significant difference between these two formulations.
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.
The susceptibility of Culex sitiens to Japanese Encephalitis (JE) virus was examined in the laboratory. Cx. sitiens became infected with JE virus on day 8 and subsequently it is able to transmit the virus when it takes a blood meal. Both parts of the experiment were carried out using artificial membrane feeding technique.
Plasmodium knowlesi a simian malaria parasite is currently affecting humans in Southeast Asia. Malaysia has reported the most number of cases and P. knowlesi is the predominant species occurring in humans. The vectors of P. knowlesi belong to the Leucosphyrus group of Anopheles mosquitoes. These are generally described as forest-dwelling mosquitoes. With deforestation and changes in land-use, some species have become predominant in farms and villages. However, knowledge on the distribution of these vectors in the country is sparse. From a public health point of view it is important to know the vectors, so that risk factors towards knowlesi malaria can be identified and control measures instituted where possible. Here, we review what is known about the knowlesi malaria vectors and ascertain the gaps in knowledge, so that future studies could concentrate on this paucity of data in-order to address this zoonotic problem.
A study was carried out to determine the distribution of cockroaches in two different housing areas with central sewerage or individual septic tanks in an urban area in Kuala Lumpur, Malaysia. Six species of cockroaches were present and of these Periplaneta americana and Periplaneta brunnea were found in greater abundance. Seventeen species of bacteria were isolated and of these Escherichia coli and Klebsiella p. pneumoniae were isolated in greatest numbers. Control measures carried out using lambda cyhalothrin showed that there was no significant difference between treated and control sites.
The residual effectiveness of 0.005mg/ml of cyhalothrin applied to cattle was determined against three species of mosquitos: Anopheles maculatus Theobald. Anopheles dirus Peyton and Harrison Mansonia uniformis Theobald. Twenty-four hour post exposure mortality and the degree of successful blood engorgement were determined by exposing mosquitos for 10 minutes to cattle. Three replicated assays were conducted and mortality determined at 1, 2, 7, 14 and 21 days after each treatment. An initial mortality of 92-94% for An. dirus and Ma. uniformis and 79% for An. maculatus was obtained. Percentage mortality declined to 10%, 18% and 31% for An. maculatus, An. dirus and Ma. uniformis respectively on day 7 post application. On day 21 post application, percentage mortality was 2-3% for the three species of mosquitos.
CDC Light traps were used to study the attractant effect of CO2 and 1-octen-3-ol on trap catches of mosquito populations at three different locations in Malaysia. There was a significant increase in the number of mosquitos caught in traps baited with CO2 and CO2 with 1-octen-3-ol. The number of mosquitos caught in the CDC light trap and in the CDC light trap baited with 1-octen-3-ol alone were very few. 1-octen-3-ol and CO2 acted synergistically in attracting significantly greater numbers of Culex tritaeniorhynchus. However Anopheles sp. were not very attracted to light traps even with attractants added to them.
Laboratory strain of the Malaysian Culex quinquefasciatus was susceptible to Wuchereria bancrofti. Thirty three percent of the Cx. quinquefasciatus that fed on W. bancrofti patient were infective after 12-14 days. There is a possibility for W. bancrofti to occur in the urban areas of the Malaysia in the near future.