Seven villages in Banggi Island, Sabah, Malaysia, were surveyed four times to evaluate the roles of local mosquitoes as vectors of malaria and Bancroftian filariasis. 11 species of Anopheles were found biting man. 53.9% of the anophelines caught were An. flavirostris, 27.1% An. balabacensis, 6% An. donaldi and 4.2% An. subpictus. Infective malaria sporozoites, probably of human origin, were found in two of 336 An. flavirostris and 12 of 308 An. balabacensis. Sporozoites, probably of a non-human Plasmodium, were found in An. umbrosus. Nine of 1001 An. flavirostris and four of 365 An. balabacensis harboured L2 or L3 filarial larvae identified as those of Wuchereria bancrofti. This is the first record of An. flavirostris as a natural vector of malaria and W. bancrofti in Sabah.
Surveillance methods for Coquillettidia crassipes were studied in an open housing estate near Kuala Lumpur using three types of traps Trinidad 10 trap, modified Lard can trap and IMR trap, each baited with chicken or pigeon. All traps attracted Cq. crassipes. There was no significant difference in the catches in the three traps. There was also no significant difference between chicken and pigeon as bait. Catches at heights of 1.5, 3, 4.5 and 6 m did not show any significant difference in density. Cq. crassipes was active at night with an early peak during the first hour of the night and a minor peak between 0100 and 0200 hours. The activity of the parous and nulliparous sections of the population was similar, except that a higher proportion of the parous females was active during the second peak compared with the nulliparous females. The parous rate was 22.3%, and the probability of survival through one day for two gonotrophic cycles was 0.711 and 0.650. The infection rate for Cardiofilaria was 29 out of 1052 (2.76%) and the infective rate (L3 larvae) was 13 out of 1052 (1.24%). 48.3% of the infected Cq. crassipes had a worm burden of more than ten larvae. One of the chickens in the traps was positive for microfilariae of Cardiofilaria four weeks after exposure as bait. Laboratory bred Cq. crassipes fed on this chicken produced infective larvae in ten days, and these were inoculated into clean chickens and pigeons. Microfilariae appeared in the chickens but not in pigeons. The adult worms recovered await identification.
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.
The role of some adult flies (Diptera: Cyclorrhapha) as carriers of helminth parasites of man was studied at four sites in Malaysia: a refuse dump, where no helminth-positive flies were detected, and in three peri-domestic situations where four species of flies carried up to three types of nematodes. The dominant fly species Chrysomya megacephala (Fabricius) carried eggs of the roundworm Ascaris lumbricoides L., the pinworm Trichuris trichiura (L.) and hookworm on the adult external body surface and in the gut lumen, in association with Bukit Lanjan aborigines. Chrysomya rufifacies (Macquart) and Sarcophaga spp. also had Ascaris lumbricoides and Trichurus trichiura eggs in their gut contents. Human helminths were not recovered from Lispe leucospila (Wiedemann), Lucilia cuprina (Wiedemann) or the housefly Musca domestica L. In an urban slum area of Kuala Lumpur city, filariform larvae identified as the hookworm Necator americanus (Stiles) occurred in the intestines of the face-fly Musca sorbens Wiedemann (22 larvae per 100 flies) and of Chrysomya megacephala (4.5 larvae per 100 flies). This concentration of apparently infective N. americanus in M. sorbens, a fly which often breeds in faeces and browses on human skin, could have transmission potential.
In Peninsular Malaysia, a large proportion of malaria cases occur in the central mountainous and forested parts of the country. As part of a study to assess remote sensing data as a tool for vector mapping, we conducted entomological surveys to determine the type of mosquitoes, their characteristics and the abundance of habitats of the vector Anopheles maculatus in malaria endemic areas in Pos Senderot. An. maculatus mosquitoes were collected from 49 breeding sites in Pos Senderot. An. maculatus preferred to breed in water pockets formed on the bank of rivers and waterfalls. The most common larval habitats were shallow pools 5.0-15.0 cm deep with clear water, mud substrate and plants or floatage. The mosquito also preferred open or partially shaded habitats. Breeding habitats were generally located at 100-400 m from the nearest human settlement. Changes in breeding characteristics were also observed. Instead of breeding in slow flowing streams, most larvae bred in small water pockets along the river margin.
Anopheles annularis is widely distributed mosquito species all over the country. An. annularis has been incriminated as a malaria vector in India, Sri Lanka, Bangladesh, Myanmar, Indonesia, Malaysia and China. In India, it has been reported to play an important role in malaria transmission as a secondary vector in certain parts of Assam, West Bengal and U.P. In Odisha and some neighbouring countries such as Sri Lanka, Nepal and Myanmar it has been recognised as a primary vector of malaria. This is a species complex of two sibling species A and B but the role of these sibling species in malaria transmission is not clearly known. An. annularis is resistant to DDT and dieldrin/HCH and susceptible to malathion and synthetic pyrethorides in most of the parts of India. In view of rapid change in ecological conditions, further studies are required on the bionomics of An. annularis and its role in malaria transmission in other parts of the country. Considering the importance of An. annularis as a malaria vector, the bionomics and its role in malaria transmission has been reviewed in this paper. In this communication, an attempt has been made to review its bionomics and its role as malaria vector. An. annularis is a competent vector of malaria, thus, due attention should be paid for its control under the vector control programmes specially in border states where it is playing a primary role in malaria transmission.
Many of the most widely spread vector-borne diseases are water related, in that the mosquito vectors concerned breed or pass part of their lifecycle in or close to water. A major reason for the study of mosquito larval ecology is to gather information on environmental variables that may determine the species of mosquitoes and the distribution of larvae in the breeding habitats. Larval surveillance studies were conducted six times between May 2008 and October 2009 in Pos Lenjang, Kuala Lipis, Pahang. Twelve environmental variables were recorded for each sampling site, and samples of mosquito larvae were collected. Larval survey studies showed that anopheline and culicine larvae were collected from 79 and 67 breeding sites, respectively. All breeding sites were classified into nine habitat groups. Culicine larvae were found in all habitat groups, suggesting that they are very versatile and highly adaptable to different types of environment. Rock pools or water pockets with clear water formed on the bank of rivers and waterfalls were the most common habitats associated with An. maculatus. Environmental variables influence the suitability of aquatic habitats for anopheline and culicine larvae, but not significantly associated with the occurrence of both larvae genera (p>0.05). This study provides information on mosquito ecology in relation to breeding habitats that will be useful in designing and implementing larval control operations.
Using the cow-baited trap (CBT) method, 1,845 Anopheles mosquitos, comprising 14 species, were caught in malaria-endemic area of Hulu Perak district, Peninsular Malaysia. The two dominant species were An. barbirostris (18.59%) and An. aconitus (18.86%). Anopheles maculatus, the main malaria vector, constituted 9.11% of the total number of mosquitos sampled. Three hundred and seventy-seven Anopheles larvae, comprising 8 species, were sampled using the North Carolina Biological Station dipper. Anopheles barbirostris larvae amounted to 64.69% of the total number of larvae; An. aconitus accounted for 10.65% of larvae. Seven habitats were identified as breeding places of Anopheles. Most species were found to breed in paddies, fishponds, and rivers. Other less popular habitats were temporary pools, mountain streams, and spring wells.
The Filariasis Control Program was established more than 30 years ago in the country and the disease is still a public health problem in some states. Since 1983, a total of 17 filariasis control teams were formed throughout the country to carry out filariasis control work. The teams conduct house and population censuses, nocturnal mass blood surveys and treatment of microscopically confirmed cases. Individual case follow-up is being carried out after 3-5 months while the locality is resurveyed after about 2-3 years. During the years 1988 to 1990, there appeared to be a decreasing trend in the number of filariasis cases detected countrywide. In 1991, brugian filariasis accounted for 92% of the cases detected. The microfilaria rate (MFR) also showed a decreasing trend countrywide for the years 1988 (0.57%) to 1990 (0.35%) but there was an increase in 1991 although it remained well below the 5% MFR targeted in the program objective, In 1991, the filariasis control teams and the district multi-purpose teams collected a total of 167, 151 blood slides out of which 871 were found to be positive for microfilaria. To determine the true endemicity of filariasis in the country, the malaria district multi-purpose teams are also utilized to assist in probe surveys in new areas of the district. Two species of filarial worms, namely Brugia malayi and Wuchereria bancrofti, and the mosquito vectors belonging to the Anopheles and Mansonia genera are involved in the transmission of filariasis in Malaysia. Monkeys and domestic cats are the reservoir hosts for the subperiodic strain of B. malayi.
In Armigeres subalbatus, 60% and 3% of the ingested Brugia pahangi microfilariae (mf) respectively migrated into the haemocoel and the thorax within 5 minutes post ingestion (p.i.). Most of the mf had migrated from the gut into the haemocoel within the first 10 minutes p.i. There was no correlation between the number of mf ingested and the migration rate though those in mosquitoes with a low mf burden tend to migrate earlier. At 24 hours p.i., 5-30% of the mf were still in the gut; 19% of these mf were immobile. At 48 hours p.i. only 2% of the mf were mobile. B. pahangi mf isolated from blood meals at 24 hours p.i., failed to develop when inoculated into Armigeres subalbatus. 54% and 73% of the mf isolated from a 24 hour old clotted blood of a B. pahangi-infected cat and fresh peripheral cat blood respectively developed into stage-1 larva. Probably mf left in the midgut at 24 hours p.i. are the young and immature worms and are physiologically incapable of penetrating the gut.
The relationship among body size (as indicated by wing length), age (as indicated by parity dissections), and malaria infection were observed in host-seeking Anopheles maculatus Theobald females collected in aboriginal villages of peninsular Malaysia. Both ELISA and salivary gland dissections were used to determine malaria infection. The wings of parous females were significantly longer than those of nulliparous females, suggesting that larger females live longer than smaller ones, and thus have a higher vectorial capacity. Body size differences were not detected between infected parous and uninfected parous females. Females infected with only oocysts were significantly larger than females infected with sporozoites. No correlation was found between the number of oocysts or sporozoites and body size in this small sample.
Studies on age groups within activity cycles, age composition and survivorship in natural populations of Mansonia in Kampung Pantai, Bengkoka Peninsula of Sabah state have been described. Early activity of 3-5 parous Ma. bonneae during the first hour after sunset was noted. Age composition of Ma. bonneae at forest shade, indoor and outdoor of house, comparative buffalo vs human bait outdoor in Kampung Pantai showed all round high parous rates ranging from 66.7 to 75.4%. Population 3-parous and older ranged from 18.8 to 26.7%. Nine of the 14 infective Ma. bonneae were 3-parous and this segment of the population was engaged in active transmission. High parous rates were observed in Ma. dives and Ma. uniformis taken in small numbers. Parous rates of Ma. bonneae taken in Kampung Delima and Kampung Taradas were also high. Estimates of daily survivorship of Ma. bonneae and Ma. dives determined by two methods were very high.
Holoendemic malaria transmission in two small isolated forest communities and a coastal village was studied by (1) all night human bait collections of Anopheles species from inside and outside houses and (2) buffalo-biting and CDC light-trapping catches during March and November 1984. During the same period thick and thin blood films were collected from the human population, and spleen rates were determined in children from two to nine years of age. Using both the immunoradiometric assay (IRMA) and the dissection technique, more sporozoite-positive infections were detected in An. balabacensis and An. flavirostris in November than in March. IRMA confirmed the presence of Plasmodium falciparum sporozoites. An average of 76.2% of the An. balabacensis population lived long enough to have reached a point where infectivity with P. falciparum was possible in November. Although fewer than five adult females bit humans per night at any time, a resident could theoretically have received more than 160 infective bites in one year. A high frequency of feeding on humans, coupled with increased anopheline life expectancy, contributed to high estimates of falciparum malaria vectorial capacity (number of infections distributed per case per day); for An. balabacensis (1.44-7.44 in March and 9.97-19.7 in November) and for An. flavirostris (0.19-5.14 in March and 6.27-15.8 in November). These high values may explain the increased malaria parasite rates obtained from at least two forest communities. Correlation between actual and calculated rates of gametocytaemia was poorest in Kapitangan due to inadequate sampling of the human population. In Banggi island, malaria is stable and holoendemic, and the population enjoys a high degree of immunity.
Traditionally, engineering and environment-based interventions have contributed to the prevention of malaria in Asia. However, with the introduction of DDT and other potent insecticides, chemical control became the dominating strategy. The renewed interest in environmental-management-based approaches for the control of malaria vectors follows the rapid development of resistance by mosquitoes to the widely used insecticides, the increasing cost of developing new chemicals, logistical constraints involved in the implementation of residual-spraying programs and the environmental concerns linked to the use of persistent organic pollutants. To guide future research and operational agendas focusing on environmental-control interventions, it is necessary to learn from the successes and failures from the time before the introduction of insecticides. The objective of this paper is to describe the experiences gained in Asia with early vector control interventions focusing on cases from the former Indian Punjab, Malaysia and Sri Lanka. The paper deals primarily with the agricultural engineering and land and water management vector control interventions implemented in the period 1900-1950. The selected cases are discussed in the wider context of environment-based approaches for the control of malaria vectors, including current relevance. Clearly, some of the interventions piloted and implemented early in the last century still have relevance today but generally in a very site-specific manner and in combination with other preventive and curative activities. Some of the approaches followed earlier on to support implementation would not be acceptable or feasible today, from a social or environmental point of view.
Anopheles gambiae females are the world's most successful vectors of human malaria. However, a fraction of these mosquitoes is refractory to Plasmodium development. L3-5, a laboratory selected refractory strain, encapsulates transforming ookinetes/early oocysts of a wide variety of Plasmodium species. Previous studies on these mosquitoes showed that one major (Pen1) and two minor (Pen2, Pen3) autosomal dominant quantitative trait loci (QTLs) control the melanotic encapsulation response against P. cynomolgi B, a simian malaria originating in Malaysia.
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.
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.
Genetic variation based on mitochondrial cytochrome c oxidase I (COI) and II (COII) sequences was investigated for three black fly nominal species, Simulium metallicum Bellardi complex, S. callidum Dyar and Shannon, and S. ochraceum Walker complex, which are vectors of human onchocerciasis from Guatemala. High levels of genetic diversity were found in S. metallicum complex and S. ochraceum complex with maximum intraspecific genetic divergences of 11.39% and 4.25%, respectively. Levels of genetic diversity of these nominal species are consistent with species status for both of them as they are cytologically complexes of species. Phylogenetic analyses revealed that the S. metallicum complex from Guatemala divided into three distinct clades, two with members of this species from several Central and South American countries and another exclusively from Mexico. The Simulium ochraceum complex from Guatemala formed a clade with members of this species from Mexico and Costa Rica while those from Ecuador and Colombia formed another distinct clade. Very low diversity in S. callidum was found for both genes with maximum intraspecific genetic divergence of 0.68% for COI and 0.88% for COII. Low genetic diversity in S. callidum might be a consequence of the result being informative of only recent population history of the species.
The nuisance bites of blackflies and transmission of Onchocerca volvulus, which causes onchocerciasis, constitutes a threat to public health and an impediment to food production in rural and riverine communities in Nigeria. The entomological profile of onchocerciasis at Adani, Nigeria, was investigated from August 2010 to January 2011 to determine the transmission of O. volvulus after 15 years of ivermectin distribution in the area. A total of 548 adult female blackflies of the Simulium damnosum complex were caught using human baits and dissected. Of this number, 248 flies were caught in the wet season (August to October), while 300 flies were caught in the dry season (November to January). The relative abundance of flies at Adani varied from 21 in December to 243 in January. The monthly catches between September and October and between December and January were significantly different. The monthly population density of the flies ranged from 0.5Flies/Man/Hour (FMH) in December to 5.5FMH in January. The diurnal biting pattern of the S. damnosum complex at the site showed a bimodal peak of activity with the evening peak being higher than the morning peak except in October when the morning peak was higher than the evening peak. The morning peaks were observed between 7.00 am and 10.00 am, whereas the evening peaks occurred between 4.00 pm and 6.00 pm. The morning and evening biting peaks in all the months were not significantly different. Nulliparous flies accounted for 75.7% of the total catch, whereas 24.3% of the flies caught were parous. The infection, infective bites and transmission of O. volvulus during the study period were zero. This study suggests that transmission of O. volvulus has been halted and the flies are presently more nuisance biters than disease vectors since no stage of O. volvulus was found in the flies dissected.
The Anopheles dirus mosquito is a primary malaria vector that transmits many species of Plasmodium parasites in Thailand and is widely spread across its geographic area. In the current study, the levels of expression of the suppressor of cytokine signaling (SOCS) gene in An. dirus mosquitoes infected with P. vivax were examined. The level of the gene's expression determined by mRNA extraction in An. dirus females (n=2,400) was studied at different times (0, 12, 24, 36, and 48 h after feeding), with different types of blood feeding (non-feeding, parasite-negative blood feeding, parasite-positive blood feeding) and in different parts of the body of mosquito samples (thorax and abdomen). The datasets were analyzed based on their relative expression ratio by the 2-ΔΔCT method and were tested for significant differences with ANOVA. The results showed that the An. dirus SOCS gene was stimulated in the abdomen 12 h and 24 h after blood feeding about three times more highly than in unfed females, with the difference being significant. At 24 h after P. vivax-infected blood feeding, the SOCS gene in the abdomen was expressed more highly than 24 h after parasite-negative blood feeding and expression was almost 36 times higher than in the control group who were not fed blood. However, in the thorax at all times after feeding and non-feeding, there was no expression of the SOCS gene. Therefore, the SOCS gene in An. dirus was most highly expressed 24 h post-feeding with a P. vivax-infected bloodmeal, which indicates that the SOCS gene in the major malaria vector in Thailand plays an important role in its immune system and its response to P. vivax infection.