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SCAR markers and multiplex PCR-based identification of isomorphic species in the Anopheles dirus complex in Southeast Asia

Manguin S, Kengne P, Sonnier L, Harbach RE, Baimai V, Trung HD, et al.

Med Vet Entomol, 2002 Mar;16(1):46-54.
PMID: 11963981

The Anopheles dirus Peyton & Harrison complex of mosquitoes (Diptera: Culicidae) comprises seven known species, including important malaria vectors in Southeast Asia. Specific identification of each species of the complex, which cannot be distinguished using morphological characters, is crucial for understanding vector ecology and implementing effective control measures. Derived from individual random amplified polymorphic DNA (RAPD) markers, sequence characterized amplified regions (SCAR) were analysed for the design of specific paired-primers. Combination of six SCAR primers resulted in the development of a simple, robust, single multiplex PCR able to identify three important malaria vectors among the four most common species (A, B, C, D) of the complex: species A from several Southeast Asian countries, species B from Perlis, Malaysia, and species C and D from Thailand.

Matched MeSH terms: Anopheles/classification*
Fulltext A report of Anopheles (Diptera: Culicidae) attracted to cow bait in a malaria endemic village in Peninsular Malaysia near the Thailand border

Rahman WA, Abu Hassan A, Adanan CR, Mohd Razha R

Southeast Asian J Trop Med Public Health, 1995 Jun;26(2):359-63.
PMID: 8629076

Twelve species of Anopheles were collected by using cow-baited net trap in a malarial endemic village in northern Peninsular Malaysia. Anopheles maculatus which is the main malaria vector with its densities were related to drought. An. aconitus, An. kochi and An. philippinensis were less susceptible to P. falciparum and P. vivax infection, and are not considered important in falciparum or vivax malaria transmission. Biting activities of An. kochi and An. vagus were primarily active after dusk and steadily declined after midnight. An. maculatus and An. aconitus showed biting activities throughout the night but An. maculatus showed two peaks of biting activity in the first half of the night.

Matched MeSH terms: Anopheles/classification
Enzyme polymorphism of the malaria vector, An. balabacensis (Diptera: Culicidae) revisited--why sample natural populations?

Hii JL, Chew M, Vun YS, Nasir M, Chang MS

Southeast Asian J Trop Med Public Health, 1988 Dec;19(4):689-701.
PMID: 3238482

Two separate observations from recent electrophoretic studies of the systematics and population genetics of laboratory-reared populations which had a long history of colonization in various laboratories, were found to be inconsistent with the present study which used wild-caught populations from East Malaysia. Reanalysis of the two data sets generally indicated a low amount of genetic variation in laboratory colonies. The latter is characterized by higher frequency of monomorphic loci, low average heterozygosity values and, in one extreme case, no variability at two loci. However, natural populations of An. balabacensis and An. leucosphyrus showed more protein variability by the use of polyacrylamide gel electrophoresis. Since laboratory-maintained mosquitoes are genetically and phenotypically different from those in the field, results of laboratory studies on the systematics and population genetics of Anopheles species complexes may be biased.

Matched MeSH terms: Anopheles/classification
Genetic variation in the malaria mosquito vector Anopheles maculatus from Peninsular Malaysia

Yong HS, Chiang GL, Loong KP, Ooi CS

Southeast Asian J Trop Med Public Health, 1988 Dec;19(4):681-7.
PMID: 3238481

Starch-gel electrophoretic studies on nine gene-enzyme systems comprising 14 loci revealed a fair level of genetic variation in two population samples of Anopheles maculatus from Peninsular Malaysia. The proportion of polymorphic loci was 0.36 for the Fort Bertau sample and 0.29 for the Gua Musang sample, while the mean heterozygosity value was 0.09 for Fort Bertau and 0.07 for Gua Musang. The values of genetic similarity (I = 0.98) and genetic distance (D = 0.02) were of the rank of geographical populations.

Matched MeSH terms: Anopheles/classification
Population cytogenetics of the malaria vector Anopheles leucosphyrus group

Baimai V

Southeast Asian J Trop Med Public Health, 1988 Dec;19(4):667-80.
PMID: 3238480

Until recently, very little was known of Anopheles species complexes and their relationships to epidemiology and malaria transmission in Southeast Asia. During the past eight years, extensive studies on the genetics of natural populations of anopheline mosquitoes in this region, involving the interdisciplinary efforts of taxonomists, operational entomologists and biologists, have revealed groups of cryptic species of Anopheles vectors, particularly the An. leucos phyrus group. This species group comprise seventeen species and two subspecies widely distributed in the forested areas of Southeast Asia. Among these species. An. dirus Peyton and Harrison, has been shown by cytogenetic and morphological studies to be a complex of at least seven isomorphic species, provisionally designated species A, B, C, D, E, F and takasagoensis, on the Southeast Asian mainland. Cytological identification of these species is based on distinct banding patterns of salivary gland polytene chromosomes as well as heterochromatin differences in mitotic karyotypes. The five species found in Thailand (A-D, F) exhibit distinct geographic distributions. Species A is widespread throughout Thailand except in the south. Species B had been found in sympatry with species C in southern Thailand and both seem to show north-south clinal geographic variation. Species D is common on the west side of southern Thailand and along the Thai-Burmese border in sympatry with species A. Species F, An. nemophilous Peyton and Ramalingam, has been found in a population at the Thai-Malaysian border in this study although it was known to be common in southern and western Thailand and Peninsular Malaysia. Species E is known only from western India. The five species found in Thailand also exhibit seasonal variation in relative abundance and different nocturnal biting cycles. Chromosomal polymorphisms have been observed in mitotic and polytene chromosomes of An. dirus A and D. Species B and C also show heterochromatin variation in the sex chromosomes, but are monomorphic for the standard sequence in polytene chromosomes. These biological characteristics of the An. dirus complex may have implications for understanding the epidemiology of malaria in Southeast Asia. Recent cytogenetic studies of wild-caught samples of An. leucosphyrus from Sumatra, Kalimantan and southern Thailand have revealed the presence of two distinct species within this taxon. Species A is widely distributed in southern Thailand, East Malaysia and Kalimantan, while species B is confined to Sumatra. The two isomorphic species are vectors of human malaria within their range of distribution.(ABSTRACT TRUNCATED AT 400 WORDS)

Matched MeSH terms: Anopheles/classification
Molecular identification of Anopheles (Cellia) sundaicus from the Andaman and Nicobar islands of India

Alam MT, Das MK, Ansari MA, Sharma YD

Acta Trop, 2006 Jan;97(1):10-8.
PMID: 16125659

Anopheles (Cellia) sundaicus (Rodenwaldt) is an important malaria vector in the Andaman and Nicobar islands of India where it breeds in freshwater as well as in brackish water. To establish the molecular identity of An. sundaicus on these islands we analyzed samples from four geographically isolated areas-Teressa, Nancowry, Car Nicobar and Katchal islands. PCR-amplification and nucleotide sequence analysis were performed for internal transcribed spacer 2 (ITS2) and domain-3 (D3) of 28S rRNA. The ITS2 region of An. sundaicus from all four islands was identical but different from An. sundaicus A of Vietnam and An. sundaicus s.s of Malaysia. Furthermore, freshwater and brackish water forms of An. sundaicus did not reveal any sequence variation. Similarly, the D3 sequences were identical among all An. sundaicus samples from the four islands. D3 sequences for a species of the Sundaicus Complex are reported here for the first time and thus could not be compared with other regional isolates of this species. In conclusion, probably only one member of the Sundaicus Complex exists on the Andaman and Nicobar islands, which breeds in freshwater as well as in brackish water and is different from the An. sundaicus A and Malaysian An. sundaicus s.s. The identification of a new sibling species of the Sundaicus Complex in these islands is significant from the viewpoint of vector control strategies.

Matched MeSH terms: Anopheles/classification*
Epidemiology of malaria in Attapeu Province, Lao PDR in relation to entomological parameters

Vythilingam I, Sidavong B, Chan ST, Phonemixay T, Vanisaveth V, Sisoulad P, et al.

Trans R Soc Trop Med Hyg, 2005 Nov;99(11):833-9.
PMID: 16112154

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.

Matched MeSH terms: Anopheles/classification*
Filaria vector competence of some Anopheles species

Zahedi M, White GB

Trop. Med. Parasitol., 1994 Mar;45(1):27-32.
PMID: 8066378

The filaria vector competence of Anopheles stephensi was compared with Brugia-susceptible Aedes aegypti Liverpool strain, An. gambiae Badagry Lagos strain and An. dirus Perlis Malaysia strain. An. stephensi ingested more Brugia pahangi microfilariae, had the highest infectivity rate and yielded more infective mosquitoes than the other two anopheline species. The overall vector competence of An. stephensi was 0.13 times that of Ae. aegypti, 0.62 times that of An. gambiae and 2.17 times that of An. dirus. However, heavy mortality among infected An. stephensi in the present investigation indicates that the filaria vectorial capacity of the mosquito might be limited epidemiologically. The relationship between filaria vector competence and mosquito foregut armature is discussed. It was observed that the relative vector competence of the three anopheline species tested was in the same order as their relative degrees of armature elaboration. The converse would be expected if foregut armatures really give partial protection to the mosquitoes against filarial infection. It is suggested that high host microfilariae density favours larval survival proportional to the degree of armature development in Anopheles (Cellia) species.

Matched MeSH terms: Anopheles/classification
The prevalence of Anopheles (Diptera: Culicidae) mosquitoes in Sekong Province, Lao PDR in relation to malaria transmission

Vythilingam I, Phetsouvanh R, Keokenchanh K, Yengmala V, Vanisaveth V, Phompida S, et al.

Trop Med Int Health, 2003 Jun;8(6):525-35.
PMID: 12791058

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.

Matched MeSH terms: Anopheles/classification
[The first monkey malaria in Turkey: a case of Plasmodium knowlesi]

Özbilgin A, Çavuş İ, Yıldırım A, Gündüz C

Mikrobiyol Bul, 2016 Jul;50(3):484-90.
PMID: 27525405

Plasmodium knowlesi is now added to the known four Plasmodium species (P.vivax, P.falciparum, P.malariae, P.ovale) as a cause of malaria in humans because of the recent increasing rate of cases reported from countries of southeastern Asia. P.knowlesi which infects macaque monkeys (Macaca fascicularis and M.nemestrina) is transmitted to humans especially by Anopheles leucosphyrus and An.hackeri mosquitos. First human cases of P.knowlesi malaria have been detected in Malaysia which have reached high numbers in recent years and also have been reported from countries of Southeast Asia such as Thailand, Philippines, Myanmar, Singapore and Vietnam. However the number of cases reported from western countries are rare and limited only within voyagers. This report is the first presentation of an imported case of P.knowlesi malaria in Turkey and aims to draw attention to the point that it could also be detected in future. A 33-year-old male patient from Myanmar who has migrated to Turkey as a refugee, was admitted to a health center with the complaints of fever with a periodicity of 24 hours, headache, fatigue, cough, sore throat, anorexia, myalgia and arthralgia. He was prediagnosed as upper respiratory tract infection, however because of his periodical fever and background in Myanmar, thick and thin blood films were prepared and sent to our laboratory for further examinations. Microscopic examination of the thin blood films revealed erythrocytic stages compatible with P.knowlesi (three large early trophozoites in an erythrocyte, three late trophozoites with compact view, and three late band-form trophozoites). Upon this, both real-time polymerase chain reaction (Rt-PCR) targeting the small subunit ribosomal RNA (SSU-rRNA) genes of Plasmodium genus and DNA sequence analysis targeting P.knowlesi rRNA gene were performed. As a result, the suspected identification of P.knowlesi by microscopy was confirmed by Rt-PCR and DNA sequencing. The patient was treated with chloroquine and primaquine combination and in the follow-up on the seventh day after the treatment, his parasitemia and symptoms had ceased. Although there were some previous reports concerning about imported patients infected with different Plasmodium species in our country, no cases of P.knowlesi have been reported. This first case presented here emphasizes the occurence of P.knowlesi malaria in Turkey hereinafter due to the increasing number of refugees.

Matched MeSH terms: Anopheles/classification
Fulltext Malaria and Anopheles mosquitos in Malaysia

Rahman WA, Che'Rus A, Ahmad AH

Southeast Asian J Trop Med Public Health, 1997 Sep;28(3):599-605.
PMID: 9561615

Until today, malaria is still one of the most important diseases in Malaysia. This is because Malaysia is located within the equatorial zone with high temperatures and humidities, usually important for the transmission of malaria. The number of malaria cases were estimated to be around 300,000 before the launching of the Malaria Eradication Program (MEP). The program was successful in reducing the numbers progressively during the 1967-1982 period. During the period 1980-1991, the highest number of malaria cases recorded for the country was 65,283 in 1989 (16,902 in Peninsular Malaysia, 47,545 in Sabah and 836 in Sarawak) whilst the lowest was 22,218 (10,069 in Peninsular Malaysia, 11,290 in Sabah and 859 in Sarawak) in 1983. In Malaysia, there are 434 species of mosquitos, representing 20 genera. Of these, 75 species are Anopheles that comprise of 2 subgenus, i.e. Anopheles and Cellia. Of the 75 species, only 9 have been reported as vectors: An. maculatus, An balabacensis, An. dirus, An. letifer An. campestris, An. sundaicus, An. donaldi, An. leucophyrus and An. flavirostris. The behavior, seasonal abundance, biting activities and breeding sites of these species are discussed.

Matched MeSH terms: Anopheles/classification
Fulltext Plasmodium knowlesi malaria an emerging public health problem in Hulu Selangor, Selangor, Malaysia (2009-2013): epidemiologic and entomologic analysis

Vythilingam I, Lim YA, Venugopalan B, Ngui R, Leong CS, Wong ML, et al.

Parasit Vectors, 2014;7:436.
PMID: 25223878 DOI: 10.1186/1756-3305-7-436

While transmission of the human Plasmodium species has declined, a significant increase in Plasmodium knowlesi/Plasmodium malariae cases was reported in Hulu Selangor, Selangor, Malaysia. Thus, a study was undertaken to determine the epidemiology and the vectors involved in the transmission of knowlesi malaria.

Matched MeSH terms: Anopheles/classification
Monkey malaria kills four humans

Galinski MR, Barnwell JW

Trends Parasitol, 2009 May;25(5):200-4.
PMID: 19345613 DOI: 10.1016/j.pt.2009.02.002

Four human deaths caused by Plasmodium knowlesi, a simian malaria species, are stimulating a surge of public health interest and clinical vigilance in vulnerable areas of Southeast Asia. We, and other colleagues, emphasize that these cases, identified in Malaysia, are a clear warning that health facilities and clinicians must rethink the diagnosis and treatment of malaria cases presumed to be caused by a less virulent human malaria species, Plasmodium malariae.

Matched MeSH terms: Anopheles/classification
Fulltext Bionomics of Anopheles latens in Kapit, Sarawak, Malaysian Borneo in relation to the transmission of zoonotic simian malaria parasite Plasmodium knowlesi

Tan CH, Vythilingam I, Matusop A, Chan ST, Singh B

Malar J, 2008;7:52.
PMID: 18377652 DOI: 10.1186/1475-2875-7-52

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.

Matched MeSH terms: Anopheles/classification
Fulltext New vectors in northern Sarawak, Malaysian Borneo, for the zoonotic malaria parasite, Plasmodium knowlesi

Ang JXD, Kadir KA, Mohamad DSA, Matusop A, Divis PCS, Yaman K, et al.

Parasit Vectors, 2020 Sep 15;13(1):472.
PMID: 32933567 DOI: 10.1186/s13071-020-04345-2

BACKGROUND: Plasmodium knowlesi is a significant cause of human malaria in Sarawak, Malaysian Borneo. Only one study has been previously undertaken in Sarawak to identify vectors of P. knowlesi, where Anopheles latens was incriminated as the vector in Kapit, central Sarawak. A study was therefore undertaken to identify malaria vectors in a different location in Sarawak.
METHODS: Mosquitoes found landing on humans and resting on leaves over a 5-day period at two sites in the Lawas District of northern Sarawak were collected and identified. DNA samples extracted from salivary glands of Anopheles mosquitoes were subjected to nested PCR malaria-detection assays. The small subunit ribosomal RNA (SSU rRNA) gene of Plasmodium was sequenced, and the internal transcribed spacer 2 (ITS2) and mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of the mosquitoes were sequenced from the Plasmodium-positive samples for phylogenetic analysis.
RESULTS: Totals of 65 anophelines and 127 culicines were collected. By PCR, 6 An. balabacensis and 5 An. donaldi were found to have single P. knowlesi infections while 3 other An. balabacensis had either single, double or triple infections with P. inui, P. fieldi, P. cynomolgi and P. knowlesi. Phylogenetic analysis of the Plasmodium SSU rRNA gene confirmed 3 An. donaldi and 3 An. balabacensis with single P. knowlesi infections, while 3 other An. balabacensis had two or more Plasmodium species of P. inui, P. knowlesi, P. cynomolgi and some species of Plasmodium that could not be conclusively identified. Phylogenies inferred from the ITS2 and/or cox1 sequences of An. balabacensis and An. donaldi indicate that they are genetically indistinguishable from An. balabacensis and An. donaldi, respectively, found in Sabah, Malaysian Borneo.
CONCLUSIONS: Previously An. latens was identified as the vector for P. knowlesi in Kapit, central Sarawak, Malaysian Borneo, and now An. balabacensis and An. donaldi have been incriminated as vectors for zoonotic malaria in Lawas, northern Sarawak.

Matched MeSH terms: Anopheles/classification*
Comparative field evaluation of residual-sprayed deltamethrin WG and deltamethrin WP for the control of malaria in Pahang, Malaysia

Rohani A, Zamree I, Lim LH, Rahini H, David L, Kamilan D

Southeast Asian J Trop Med Public Health, 2006 Nov;37(6):1139-48.
PMID: 17333767

The bioefficacy of indoor residual-sprayed deltamethrin wettable granule (WG) formulation at 25 mg a.i./m2 and 20 mg a.i./m2 for the control of malaria was compared with the current dose of 20 mg/m2 deltamethrin wettable powder (WP) in aboriginal settlements in Kuala Lipis, Pahang, Malaysia. The malaria vector has been previously identified as Anopheles maculatus. The assessment period for the 20 mg/m2 dosage was six months, but for the 25 mg/m2 dosage, the period was 9 months. Collections of mosquitoes using the bare-leg techniques were carried out indoors and outdoors from 7:00 PM to 7:00 AM. All mosquitoes were dissected for sporozoites and parity. Larval collections were carried out at various locations to assess the extent and distribution of breeding of vectors. A high incidence of human feeds was detected during May 2005 and a low incidence during January 2005 for all the study areas. Our study showed that deltamethrin WG at 25 mg/m2 suppressed An. maculatus biting activity. More An. maculatus were caught in outdoor landing catches than indoor landing catches for all the study areas. The results indicate that 25 mg/m2 WG is good for controlling malaria for up to 9 months. Where residual spraying is envisaged, the usual two spraying cycles per year with 20 mg/m2 deltamethrin may be replaced with 25 mg/m2 deltamethrin WG every 9 months.

Matched MeSH terms: Anopheles/classification

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