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Indra A/P Vythilingam


indra.vythilingam@gmail.com

Journal articles

2012
A Jiram, I Vythilingam, Y NoorAzian, Y Yusof, A Azahari, M Y Fong (2012)  Entomologic investigation of Plasmodium knowlesi vectors in Kuala Lipis, Pahang, Malaysia   Malaria Journal 11: 1.  
Abstract: BACKGROUND: xD;The first natural infection of Plasmodium knowlesi in humans was recorded in 1965 in peninsular Malaysia. Extensive research was then conducted and it was postulated that it was a rare incident and that simian malaria will not be easily transmitted to humans. However, at the turn of the 21st century, knowlesi malaria was prevalent throughout Southeast Asia and is life threatening. Thus, a longitudinal study was initiated to determine the vectors, their seasonal variation and preference to humans and macaques. xD;METHODS: xD;Monthly mosquito collections were carried out in Kuala Lipis, Pahang, peninsular Malaysia, using human-landing collection and monkey-baited traps at ground and canopy levels. All mosquitoes were identified and all anopheline mosquitoes were dissected and the gut and gland examined for oocysts and sporozoites. Nested polymerase chain reaction (PCR) was conducted on positive samples, followed by sequencing of the csp gene. xD;RESULTS AND DISCUSSION: xD;Anopheles cracens was the predominant mosquito biting humans as well as the macaques. It comprised 63.2% of the total collection and was the only species positive for sporozoites of P. knowlesi. It was exophagic and did not enter houses. Besides An. cracens, Anopheles kochi was also found in the monkey-bait trap. Both species preferred to bite monkeys at ground level compared to canopy. xD;CONCLUSION: xD;Anopheles cracens, which belongs to the Dirus complex, Leucosphyrus subgroup, Leucosphyrus group of mosquitoes, has been confirmed to be the only vector for this site from Pahang during this study. It was the predominant mosquito at the study sites and with deforestation humans and villages are entering deeper in the forests, and nearer to the mosquitoes and macacques. The close association of humans with macaques and mosquitoes has led to zoonotic transmission of malaria.
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I Vythilingam (2012)  Plasmodium knowlesi and Wuchereria bancrofti : Their Vectors and Challenges for the Future   Frontiers in Physiology 3:  
Abstract: 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.
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2011
C H Tan, P S J Wong, M Z I Li, I Vythilingam, L C Ng (2011)  Evaluation of the Dengue NS1 Ag Strip® for Detection of Dengue Virus Antigen in Aedes aegypti (Diptera : Culicidae)   Vector-Borne and Zoonotic Diseases 11: 6. 789-792  
Abstract: Abstract xD;Dengue fever is currently one of the most important mosquito-borne diseases that affect humans. With neither vaccines nor treatment available, prevention of the disease relies heavily on surveillance and control of mosquito vectors. In the present study, we have evaluated and showed the potential use of the Dengue NS1 Ag Strip® for the detection of dengue virus (DENV) in Aedes aegypti. Initial results showed that the sensitivity of the test kit in detecting DENV in wild-caught mosquitoes is comparable to that of real-time reverse transcriptase–polymerase chain reaction. The detection of naturally infected Ae. aegypti with the NS1 rapid test kit in our dengue cluster investigation further illustrates its potential use for surveillance of DENV in wild mosquito populations. The kit can easily be used in a simple field station, and minimal training is required. The results can be obtained in less than an hour. Employment of the kit in the field could help guide mosquito control operations in the prioritization of resources in controlling the transmission of DENV. In this study the potential of the kit for field surveillance of infected dengue vectors, which are epidemiologically important, has been demonstrated.
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2010
P Van den Eede, I Vythilingam, D T Ngo, V H Nguyen, X H Le, U D'Alessandro, A Erhart (2010)  Plasmodium knowlesi malaria in Vietnam : some clarifications   Malar J 9: 20.  
Abstract: A recently published comment on a report of Plasmodium knowlesi infections in Vietnam states that this may not xD;accurately represent the situation in the study area because the PCR primers used may cross-hybridize with Plasmodium xD;vivax. Nevertheless, P. knowlesi infections have been confirmed by sequencing. In addition, a neighbourjoining xD;tree based on the 18S S-Type SSUrRNA gene shows that the Vietnamese samples clearly cluster with the P. xD;knowlesi isolates identified in Malaysia and are distinct from the corresponding P. vivax sequences. All samples xD;came from asymptomatic individuals who did not consult for fever during the months preceding or following the xD;survey, indicating that asymptomatic P. knowlesi infections occur in this population, although this does not exclude xD;the occurrence of symptomatic cases. Large-scale studies to determine the extent and the epidemiology of P. xD;knowlesi malaria in Vietnam are further needed.
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F Anderios, A NoorRain, I Vythilingam (2010)  < i> In vivo study of human< i> Plasmodium knowlesi in< i> Macaca fascicularis   Experimental parasitology 124: 2. 181-189  
Abstract: Abstract xD;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 μL of blood for asexual stage and 88–264 parasites per μL of blood for sexual stage. Periodicity analysis adopted from microfilaria periodicity technique of asexual stage showed that the parasitemia peak at 17:39 h 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.
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N Lee-Ching, L Kim-Sung, T Cheong-Huat, O Peng-Lim, L P Sai-Gek, L Raymond, L Yee-Ling, S Suhana, C Pei-Pei, W Kit-Yin, Vythilingam Indra (2010)  Entomologic and molecular investigation into Plasmodium vivax transmission in Singapore, 2009   Malaria Journal 9: 1.  
Abstract: Background xD; xD;Singapore has been certified malaria free since November 1982 by the World Health Organization and despite occasional local transmission, the country has maintained the standing. In 2009, three clusters of malaria cases were reported in Singapore. xD; xD;Methods xD; xD;Epidemiological, entomological and molecular studies were carried out to investigate the three clusters, namely Mandai-Sungei Kadut, Jurong Island and Sembawang. xD; xD;Results xD; xD;A total of 29 malaria patients, with no recent travel history, were reported in the three clusters. Molecular analysis based on the msp3α and msp1 genes showed two independent local transmissions: one in Mandai-Sungei Kadut and another in Sembawang. Almost all cases within each cluster were epidemiologically linked. In Jurong Island cluster, epidemiological link remains uncertain, as almost all cases had a unique genetic profile. Only two cases shared a common profile and were found to be linked to the Mandai-Sungei Kadut cluster. Entomological investigation found Anopheles sinensis to be the predominant Anopheline in the two areas where local transmission of P. vivax was confirmed. Anopheles sinensis was found to be attracted to human bait and bites as early as 19:45 hrs. However, all Anopheles mosquitoes caught were negative for sporozoites and oocysts by dissection. xD; xD;Conclusion xD; xD;Investigation of P. vivax cases from the three cluster areas confirmed the occurrence of local transmission in two areas. Although An. sinensis was the predominant Anopheline found in areas with confirmed transmission, the vector/s responsible for the outbreaks still remains cryptic.
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I Vythilingam (2010)  Review Paper Plasmodium knowlesi in humans : a review on the role of its vectors in Malaysia   Tropical Biomedicine 27: 1. 1-12  
Abstract: 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.
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2009
P V Eede, H N Van, C Van Overmeir, I Vythilingam, T N Duc, L X Hung, H N Manh, J AnnĂ©, U D'Alessandro, A Erhart (2009)  Human Plasmodium knowlesi infections in young children in central Vietnam   Malaria journal 8: 1.  
Abstract: Background xD;Considering increasing reports on human infections by Plasmodium knowlesi in Southeast Asian countries, blood samples collected during two large cross-sectional malariometric surveys carried out in a forested area of central Vietnam in 2004 and 2005 were screened for this parasite. xD; xD;Methods xD;Blood samples collected at the 2004 survey and positive for Plasmodium malariae were randomly selected for PCR analysis detecting P. knowlesi. Blood samples collected in 2005 from the same individuals were screened again for P. knowlesi. Positive samples were confirmed by sequencing. Family members of positive cases who participated in both surveys were also screened. xD; xD;Results xD;Ninety-five samples with P. malariae mono- or mixed infections identified by species-specific PCR were screened for P. knowlesi. Among the five (5.2%) positive samples by PCR, three were confirmed to be P. knowlesi infections by sequencing, two young children (<5 years old) and a young man, all asymptomatic at the time of the survey and for the next six months after the survey. One of the two children was still positive one year later. No infection was found among the family members. xD; xD;Conclusion xD;Plasmodium knowlesi infections in humans can be found in central Vietnam. A small child was positive for P. knowlesi in both surveys at one year interval, though it is unclear whether it was the same or a new infection.
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2008
C H Tan, I Vythilingam, A Matusop, S T Chan, B Singh (2008)  Bionomics of Anopheles latens in Kapit, Sarawak, Malaysian Borneo in relation to the transmission of zoonotic simian malaria parasite Plasmodium knowlesi   Malar J 7: 1.  
Abstract: Background xD;A large focus of human infections with Plasmodium knowlesi, a simian parasite naturally found in long-tailed and pig-tailed macaques was discovered in the Kapit Division of Sarawak, Malaysian Borneo. A study was initiated to identify the vectors of malaria, to elucidate where transmission is taking place and to understand the bionomics of the vectors in Kapit. xD; xD;Methods xD; xD;Three different ecological sites in the forest, farm and longhouse in the Kapit district were selected for the study. Mosquitoes were collected by human landing collection at all sites and at the forest also by monkey-baited-traps situated on three different levels. All mosquitoes were identified and salivary glands and midguts of anopheline mosquitoes were dissected to determine the presence of malaria parasites. xD; xD;Results and Discussions xD; xD;Over an 11-month period, a total of 2,504 Anopheles mosquitoes comprising 12 species were caught; 1,035 at the farm, 774 at the forest and 425 at the longhouse. Anopheles latens (62.3%) and Anopheles watsonii (30.6%) were the predominant species caught in the forested ecotypes, while in the farm Anopheles donaldi (49.9%) and An. latens (35.6%) predominated. In the long house, An. latens (29.6%) and An. donaldi (22.8%) were the major Anopheline species. However, An. latens was the only mosquito positive for sporozoites and it was found to be attracted to both human and monkey hosts. In monkey-baited net traps, it preferred to bite monkeys at the canopy level than at ground level. An. latens was found biting early as 18.00 hours. xD; xD;Conclusion xD; xD;Anopheles latens is the main vector for P. knowlesi malaria parasites in the Kapit District of Sarawak, Malaysian Borneo. The study underscores the relationship between ecology, abundance and bionomics of anopheline fauna. The simio-anthropophagic and acrodendrophilic behaviour of An. latens makes it an efficient vector for the transmission of P. knowlesi parasites to both human and monkey hosts. xD; xD;Go to: xD;Background xD;Malaria parasites in Peninsular Malaysian monkeys were first reported in 1908 [1], but only gained prominence in the 1960's after the accidental discovery [2] that Plasmodium cynomolgi could be transmitted to humans via mosquito bites in the laboratory. This stimulated interest at a time when the Malaria Eradication Programme was initiated by the World Health Organization [3] and it was important to determine if malaria was a zoonosis. Therefore, extensive studies were carried out in Peninsular Malaysia to determine the distribution, prevalence and species of malaria parasites in monkeys and apes and the natural vectors of monkey malaria parasites [4-7]. Instead of uncovering human cynomolgi malaria infections, Plasmodium knowlesi was the first simian malaria parasite found to be infecting humans in nature. The first case was reported in 1965 from the state of Pahang [8], Peninsular Malaysia, followed by a second case five years later acquired from Johore, Peninsular Malaysia [9]. It was postulated that P. knowlesi could be transmitted from monkeys to man and laboratory studies proved that it was possible [10]. However, a large scale study that was initiated in Pahang to investigate whether malaria was a zoonosis, by a group of American and local researchers based at the Institute for Medical Research in Kuala Lumpur, Peninsular Malaysia, concluded that simian malaria in humans was an extremely rare event [11,12]. This was based on their studies in which they collected blood samples from more than 1,100 local residents, pooled the samples and injected them into rhesus monkeys and none of the monkeys contracted malaria. However, in 2004 a large focus of human P. knowlesi infection was reported in the Kapit Division of Sarawak [13]. In that study 71.6% (101/141) of human malaria cases at Kapit Hospital which had been identified by microscopy as single Plasmodium malariae infections were actually P. knowlesi and other non-P. malariae species by nested polymerase chain reaction (PCR) assays. Plasmodium knowlesi is naturally found in long-tailed macaques (Macaca fascicularis), pig-tailed macaques (Macaca nemestrina) [14] and banded leaf monkeys (Presbytis malalophos) [15,16]. Since transmission of this zoonotic parasite to humans is occurring in the Kapit Division of Sarawak, Malaysian Borneo, it is important to identify the vectors so that appropriate measures can be planned and initiated to control the spread of simian malaria in humans. xD; xD;Numerous studies on vectors of human malaria have been carried out in Sarawak [17-20] but none of these have been undertaken in the Kapit District of Sarawak. Several anopheline species incriminated or suspected to transmit monkey malaria parasites in Peninsular Malaysia, such as Anopheles latens, Anopheles balabacensis are also present in Sarawak [21]. Hence, the vectorial status of the anopheline species present in the Kapit Division needs to be ascertained in order to determine if they are competent vectors of simian malaria parasites. Furthermore, the epidemiological data of P. knowlesi infection in humans revealed that infections occur primarily in adults and no clustering of cases occurred within communities that live in communal longhouses, which suggests transmission of P. knowlesi to humans occurred away from the vicinity of the longhouses [13]. xD; xD;Thus the objectives of this study were to determine the vectors of P. knowlesi and other simian malaria parasites in the Kapit Division of Sarawak; and to study the dynamics of these vectors in different ecological sites in order to elucidate the most likely place where transmission was taking place. Preliminary results of this study were reported where An. latens was incriminated as the vector for P. knowlesi [22]. Here detailed results of this eleven-month study on vectors of malaria and their bionomics in the Kapit district of Sarawak, Malaysian Borneo is presented.
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I Vythilingam, Y M NoorAzian, T C Huat, A I Jiram, Y M Yusri, A H Azahari, I NorParina, A NoorRain, S LokmanHakim (2008)  Plasmodium knowlesi in humans, macaques and mosquitoes in peninsular Malaysia   Parasit Vectors 1: 1.  
Abstract: Background Since a large focus of human infection with Plasmodium knowlesi , a simian malaria parasite naturally found in long-tailed and pig tailed macaques, was reported in Sarawak, Malaysian Borneo, it was pertinent to study the situation in peninsular Malaysia. A study was thus initiated to screen human cases of Plasmodium malariae using molecular techniques, to determine the presence of P. knowlesi in non- human primates and to elucidate its vectors. Methods Nested polymerase chain reaction (PCR) was used to identify all Plasmodium species present in the human blood samples sent to the Parasitology laboratory of Institute for Medical Research. At the same time, non-human primates were also screened for malaria parasites and nested PCR was carried out to determine the presence of P. knowlesi . Mosquitoes were collected from Pahang by human landing collection and monkey-baited-traps situated on three different levels. All mosquitoes were identified and salivary glands and midguts of anopheline mosquitoes were dissected to determine the presence of malaria parasites and nested PCR was carried out on positive glands. Sequencing of the csp genes were carried on P. knowlesi samples from humans, monkeys and mosquitoes, positive by PCR. Results and Discussion Plasmodium knowlesi was detected in 77 (69.37%) of the 111 human samples, 10 (6.90%) of the 145 monkey blood and in 2 (1.7%) Anopheles cracens . Sequence of the csp gene clustered with other P. knowlesi isolates. Conclusion Human infection with Plasmodium knowlesi is occurring in most states of peninsular Malaysia. An. cracens is the main vector. Economic exploitation of the forest is perhaps bringing monkeys, mosquitoes and humans into increased contact. A single bite from a mosquito infected with P. knowlesi is sufficient to introduce the parasite to humans. Thus, this zoonotic transmission has to be considered in the future planning of malaria control.
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2007
M Y Noor Azian, Y M San, C C Gan, M Y Yusri, Y Nurulsyamzawaty, A H Zuhaizam, M N Maslawaty, I Norparina, I Vythilingam (2007)  Prevalence of intestinal protozoa in an aborigine community in Pahang, Malaysia   Tropical Biomedicine 24: 1. 55-62  
Abstract: The objective was to estimate the prevalence of intestinal protozoa among the aborigines and to determine the problems regarding the infection. The study was carried out in January 2006 in Pos Senderut, Pahang, Malaysia. Samples of faeces were collected from children and adults and these were fixed in PVA and trichrome staining was carried out. From the 130 individuals studied, 94 (72.3%) were positive with at least one intestinal protozoa. Nine intestinal protozoa namely Blastocystis hominis, Giardia lamblia, Entamoeba histolytica, Entamoeba coli, Endolimax nana, Entamoeba hartmani, Entamoeba polecki, Iodamoeba butschlii and Chilomastix mesnili were detected. The prevalent species were B. hominis (52.3%), followed by G. lamblia (29.2%), E. coli (26.2%) and E. histolytica (18.5%). The other species ranged from 1.5 to 10.8%. Among the positive samples, mixed infection with E. histolytica and G. lamblia was 3.8%, E. histolytica and B. hominis was 15.4%, G. lamblia and B. hominis was 17.7%. Triple infection of E. histolytica, G. lamblia and B. hominis was 3.1%. The infection was more prevalent in children below 10 years age group (45.4%) and lowest in the age above 60 years (3.8%). The high prevalence was attributable to poor environmental management, poor personal hygiene and lack of health education.
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I Vythilingam, J Jeffery, R E Harbach, S T Chan, C H Tan, M Asmad (2007)  Anopheles (Cellia) Kokhani, N. SP.(Diptera Culicidae) from Kapit, Sarawak, East Malaysia   Proceedings of the Entomological Society of Washington 109: 2. 463-468  
Abstract: Anopheles kokhani Vythilingam, Jeffery, and Harbach, n. sp., a member of the Neomyzomyia Series of subgenus Cellia Theobald (Diptera: Culicidae) is described from females collected in Sarawak, Malaysia, during studies to identify the mosquitoes responsible for transmitting simian malaria to humans. The species is distinguished from sympatric members of the Neomyzomyia Series, including An. saungi Colless, An. stookesi Colless, An. tessellatus Theobald, and An. watsonii (Leicester), with which it shares one or more key features. The wing and legs of the new species are illustrated.
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I Dusfour, J Blondeau, R E Harbach, I Vythilingham, V Baimai, H D Trung, T Sochanta, M J Bangs, S Manguin (2007)  Polymerase chain reaction identification of three members of the Anopheles sundaicus (Diptera : Culicidae) complex, malaria vectors in Southeast Asia   Journal of medical entomology 44: 5. 723-731  
Abstract: Anopheles sundaicus s.l., a major malaria vector taxon, occurs primarily along coastal areas and on islands in Southeast Asia. Our previous studies using cytochrome oxidase I, cytochrome-b, and internal transcribed spacer 2 markers discriminated three allopatric species: An. sundaicus s.s. in northern Borneo, An. epiroticus in Southeast Asia, and An. sundaicus E on Sumatra and Java, Indonesia. Morphological comparisons of three developmental stages did not reveal unique diagnostic characters that could reliably distinguish the three species. Therefore, we developed a multiplex polymerase chain reaction (PCR) assay based on two mitochondrial DNA markers to unambiguously identify them. This PCR was tested on 374 specimens from 24 different geographical populations, expanding our knowledge of the distribution of these species.
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C Walton, P Somboon, S M O’loughlin, S Zhang, R E Harbach, Y M Linton, B Chen, K Nolan, S Duong, M Y Fong, I Vythilingam (2007)  Genetic diversity and molecular identification of mosquito species in the< i> Anopheles maculatus group using the ITS2 region of rDNA   Infection, Genetics and Evolution 7: 1. 93-102  
Abstract: The species diversity and genetic structure of mosquitoes belonging to the Anopheles maculatus group in Southeast Asia were investigated using the internal transcribed spacer 2 (ITS2) of ribosomal DNA (rDNA). A molecular phylogeny indicates the presence of at least one hitherto unrecognised species. Mosquitoes of chromosomal form K from eastern Thailand have a unique ITS2 sequence that is 3.7% divergent from the next most closely related taxon (An. sawadwongporni) in the group. In the context of negligible intraspecific variation at ITS2, this suggests that chromosomal form K is most probably a distinct species. Although An. maculatus sensu stricto from northern Thailand and southern Thailand/peninsular Malaysia differ from each other in chromosomal banding pattern and vectorial capacity, no intraspecific variation was observed in the ITS2 sequences of this species over this entire geographic area despite an extensive survey. A PCR-based identification method was developed to distinguish five species of the group (An. maculatus, An. dravidicus, An. pseudowillmori, An. sawadwongporni and chromosomal form K) to assist field-based studies in northwestern Thailand. Sequences from 187 mosquitoes (mostly An. maculatus and An. sawadwongporni) revealed no intraspecific variation in specimens from Thailand, Cambodia, mainland China, Malaysia, Taiwan and Vietnam, suggesting that this identification method will be widely applicable in Southeast Asia. The lack of detectable genetic structure also suggests that populations of these species are either connected by gene flow and/or share a recent common history.
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2006
I Vythilingam, C H Tan, M Asmad, S T Chan, K S Lee, B Singh (2006)  Natural transmission of Plasmodium knowlesi to humans by Anopheles latens in Sarawak, Malaysia   Transactions of the Royal Society of Tropical Medicine and Hygiene 100: 11.  
Abstract: Four species of malaria parasites are known to infect humans. A fifth species, Plasmodium knowlesi, has been reported to infect humans in Malaysian Borneo. Here we report for the first time the incrimination of Anopheles latens as the vector of P. knowlesi among humans and monkeys in Sarawak, Malaysia.
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I VYTHILINGAM, B SIDAVONG, C S THIM, T PHONEMIXAY, S PHOMPIDA, J JEFFERY (2006)  Species Composition of Mosquitoes of Attapeu Province, Lao People's Democratic Republic   Journal of the American Mosquito Control Association 22: 1. 140-143  
Abstract: A list of mosquitoes collected from Lao People's Democratic Republic (Lao PDR) from May 2002 to August 2004 is presented. A total of 8,945 mosquitoes belonging to 14 genera and 57 species was collected. The genera recorded were Aedes, Anopheles, Armigeres, Coquillettidia, Culex, Diceromyia, Downsiomyia, Fredwardsius, Mansonia, Neomelaniconion, Ochlerotatus, Paraedes, Stegomyia, and Verrallina. Seventeen species are first reports for LAO PDR.
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2005
I Vythilingam, B Sidavong, S T Chan, T Phonemixay, V Vanisaveth, P Sisoulad, R Phetsouvanh, S L Hakim, S Phompida (2005)  Epidemiology of malaria in Attapeu Province, Lao PDR in relation to entomological parameters   Transactions of the Royal Society of Tropical Medicine and Hygiene 99: 11. 833-839  
Abstract: Surveys were conducted in malaria-endemic villages in the southern province of Attapeu, Lao PDR during various seasons over a 3-year period. All-night mosquito landing collections, blood surveys and a case-control study were conducted. Plasmodium falciparum was the predominant species, and slide positivity rates were higher during the transition/dry season compared with the wet season. Anopheles dirus A was found to be the primary vector, and sporozoite rates were highest during the transition/dry season. Anopheles dirus was found to be endophagic and endophilic. Not using insecticide-treated bed nets, houses close to breeding sites and sleeping away from home were risk factors associated with malaria.
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I Vythilingam, B M Luz, R Hanni, T S Beng, T C Huat (2005)  Laboratory and field evaluation of the insect growth regulator pyriproxyfen (Sumilarv 0.5 G) against dengue vectors   Journal of the American Mosquito Control Association 21: 3. 296-300  
Abstract: The insect growth regulator pyriproxyfen was tested against Aedes aegypti at 0.01 and 0.02 mg of active ingredient (AI) per liter of water in 60-liter earthern jars. Both concentrations provided 100% control for 4 months. In additional experiments where 10 liters of water were replaced fortnightly, 100% control was still obtained over 4 months with 0.02 mg AI/liter and greater than 93-100% control was obtained over 4 months with 0.01 mg AI/liter. In less-controlled field-trial conditions, pyriproxyfen at a dosage of 0.02 mg AI/liter provided 100% control for 10 wk against Aedes albopictus even though water was replaced either daily or weekly. Although the activity of pyriproxyfen declines after 10 wk, those tests in the plastic tubs showed much higher levels of sustained residual activity compared to those in the earthern jars. Pyriproxyfen did not have an impact on nontarget organisms.
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I Vythilingam, S T Chan, C Shanmugratnam, H Tanrang, K H Chooi (2005)  The impact of development and malaria control activities on its vectors in the Kinabatangan area of Sabah, East Malaysia   Acta tropica 96: 1. 24-30  
Abstract: A study was carried out from July 2001 until January 2003 in the Kinabatangan area of Sabah, part of Borneo island, where malaria used to be mesoendemic. Vector surveys determined that Plasmodium falciparum was the predominant species and Anopheles balabacensis the primary vector. Malaria cases have dropped drastically over the years but P. falciparum is still predominant. In the present study, Anopheles donaldi was the predominant species and was positive for sporozoites. Although An. balabacensis was present, none were infective. An. donaldi bite more outdoors than indoors and have a peak biting time from 18:00 to 19:00 h when most people are still out of their homes. An integrated malaria control programme along with area development has helped in the control of malaria and its vector.
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I Vythilingam, T C Huat, N W Ahmad (2005)  Research Note Transmission potential of Wuchereria bancrofti by Culex quinquefasciatus in urban areas of Malaysia   Tropical Biomedicine 22: 1. 83-85  
Abstract: 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.
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2003
I Vythilingam, R Phetsouvanh, K Keokenchanh, V Yengmala, V Vanisaveth, S Phompida, S L Hakim (2003)  The prevalence of Anopheles (Diptera : Culicidae) mosquitoes in Sekong Province, Lao PDR in relation to malaria transmission   Tropical Medicine & International Health 8: 6. 525-535  
Abstract: A longitudinal study was conducted on the prevalence of Anopheles in three malaria endemic villages in Sekong province, in the southern region of Lao PDR, from August 2000 to October 2001. All night, human landing collections took place in August and October 2000 and April and October 2001, and blood smears were taken for malaria parasites during the same period. Mosquitoes were tested for sporozoite antigen using enzyme-linked immunosorbent assay. In August 2000 (wet season) and April 2001 (dry season) the ovaries of the mosquitoes were examined for parity. A total of 16 species of Anopheles were caught in the study sites of which An. dirus A, An. maculatus sl and An. jeyporiensis were positive for sporozoites. The entomological inoculation rate (EIR) ranged from 0.06 to 0.25. There was a good correlation between EIR and vectorial capacity in the wet season, especially in Pai Mai where the prevalence of malaria was also high during the wet seasons (11.8 and 10.53). An. dirus A showed ambivalence in their choice of feeding as approximately 50% attacked man indoors and an equal proportion outdoors. An dirus A was the main vector in Pai Mai. The parous rate did not significantly differ between the wet and dry season, although it was higher in the dry season. In Takaio the parasite prevalence ranged from 8.7% (dry season) to 37.1% (wet season) and An. jeyporiensis was the vector, and the risk of infection was 0.85 in the dry season while 0.99 in the wet season. In Toumgno An. maculatus sl was the vector and infection was found only in August and October 2000. However, malaria prevalence ranged from 9.69 to 20.4% and was equally high in the dry season. Cattle were also present close to the houses in all the villages and this might be a contributory factor in the prevalence of malaria.
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2002
I Vythilingam, S B Tan, M Krishnasamy (2002)  Short communication : Susceptibility of Culex sitiens to Japanese encephalitis virus in peninsular Malaysia   Tropical Medicine & International Health 7: 6. 539-540  
Abstract: 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.
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1998
I Vythilingam, L Boaz, N Wa (1998)  Detection of Brugia malayi in mosquitoes by the polymerase chain reaction   JOURNAL-AMERICAN MOSQUITO CONTROL ASSOCIATION 14: 3. 243-247  
Abstract: 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.
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1997
H Tsuchie, K Oda, I Vythilingam, R Thayan, B Vijayamalar, M Sinniah, J Singh, T Wada, H Tanaka, T Kurimura (1997)  Genotypes of Japanese encephalitis virus isolated in three states in Malaysia   American Journal of Tropical Medicine and Hygiene 56: 2. 153-158  
Abstract: Two hundred forty nucleotides from the pre-membrane gene region of 12 Japanese encephalitis virus (JEV) strains isolated from three different regions of Malaysia from 1993 to 1994 were sequenced and compared with each other and with the JEV strains from different geographic areas in Asia. These 12 Malaysian isolates were classified into two genotypes. The four JEV strains isolated from Sarawak in 1994 and the four JEV strains isolated from Sepang, Selangor in 1993 were classified into one genotype that included earlier isolated strains from Malaysia (JE-827 from Sarawak in 1968 and WTP/70/22 from Kuala Lumpur in 1970). The four JEV strains from Ipoh, Perak in 1994 were classified into another genotype that included JEV strains isolated from northern Thailand and Cambodia. In an earlier report, 10 JEV strains from Sabak Bernam, Selangor in 1992 were classified into the largest genotype that included strains isolated in temperate regions such as Japan, China, and Taiwan. The data indicate that at least three genotypes of JEV have been circulating in Malaysia.
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I Vythilingam, K Oda, S Mahadevan, G Abdullah, C S Thim, C C Hong, B Vijayamalar, M Sinniah, A Igarashi (1997)  Abundance, parity, and Japanese encephalitis virus infection of mosquitoes (Diptera : Culicidae) in Sepang District, Malaysia   Journal of medical entomology 34: 3. 257-262  
Abstract: A 2-yr study of Japanese encephalitis (JE) virus in Sepang District, Selangor, Malaysia, was carried out to identify the mosquito vectors and to determine their seasonal abundance, parity, and infection rates. In total, 81,889 mosquitoes belonging to 9 genera and > 50 species were identified from CDC trap collections augmented with dry ice during 1992 and 1993. Culex tritaeniorhynchus Giles and Culex gelidus Giles were the most abundant species, and both increased in numbers with increases in rainfall. Overall, 45 JE virus isolations were made from 7 species-Cx. tritaeniorhynchus (24), Cx. gelidus (12), Culex fuscocephala Theobald (2), Aedes butleri Theobald (4), Culex quinquefasciatus Say (1), Aedes lineatopennis Ludlow (1), and Aedes (Cancraedes) sp. (1). Based on elevated abundance and JE infection rates, Cx. tritaeniorhynchus appears to be the most important vector of JE virus in Sepang
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1995
S L Hakim, I Vythilingam, M I Marzukhi, J W Mak (1995)  Single-dose diethylcarbamazine in the control of periodic brugian filariasis in Peninsular Malaysia   Transactions of the Royal Society of Tropical Medicine and Hygiene 89: 6. 686-689  
Abstract: The study compared the effectiveness of a single dose of diethylcarbamazine (DEC) (6mg/kg) with the standard regimen of 6 doses (total 36mg/kg) in mass chemotherapy for the control of brugian filariasis. Mass chemotherapy with single-dose DEC was instituted in one area and standard dose in the other and treatment was repeated after one year. Parasitplogical surveys were conducted before, and 3, 7 and 12 months after treatment. Pretreatment characteristics were not significantly different between the 2 treatment areas. There was a significant reduction in microfilaraemia prevalence rate from 24.7% to 14.7% at 12 months and to 6.8% at 19 months in the single dose area and from 22.8% to 9.6% at 12 months and to 2.7% at 19 months with the standard dose. Maximum reduction was at 7 months after treatment with both regimens. There was also significant progressive reduction in mean microfilarial density from 4.39 ± 20.37 to 0.89 ± 4.16 per 60 μL in the single-dose area and from 4.43 ± 17.31 to 0.75 ± 4.95 per 60 μL in the standard dose area. There was a greater reduction of both microfilarial prevalence and density using the standard regimen but it was not statistically significant. Thus, a single dose of DEC is as effective as the standard dose in controlling periodic brugian filariasis.
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I Vythilingam, K Oda, T K Chew, S Mahadevan, B Vijayamalar, K Morita, H Tsuchie, A Igarashi (1995)  Isolation of Japanese encephalitis virus from mosquitoes collected in Sabak Bernam, Selangor, Malaysia in 1992   Journal of the American Mosquito Control Association 11: 1.  
Abstract: 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.
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