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Bionomic Status of Anopheles Epiroticus Linton & Harbach, A

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Bionomic Status of Anopheles Epiroticus Linton & Harbach, A BIONOMIC STATUS OF AN. EPIROTICUS BIONOMIC STATUS OF ANOPHELES EPIROTICUS LINTON & HARBACH, A COASTAL MALARIA VECTOR, IN RAYONG PROVINCE, THAILAND Suchada Sumruayphol1, Chamnarn Apiwathnasorn1, Narumon Komalamisra1, Jiraporn Ruangsittichai1, Yudthana Samung1 and Porntip Chavalitshewinkoon-Petmitr2 1Department of Medical Entomology, 2Department of Protozoology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand Abstract. A longitudinal entomological survey was conducted to provide in-depth information on An. epiroticus and determine whether ecological and entomologi- cal factors could influence malaria transmission in Rayong Province, Thailand. The mosquitoes were collected monthly from May 2007 to April 2008 by human landing catch technique from 6:00-12:00 PM for 2 consecutive nights, at 3 collection sites. A total of 3,048 mosquitoes within 5 species were captured: An. epiroticus, Culex quinquefasciatus Say, Cx. sitiens Wiedemann, Aedes aegypti (L.) and Ae. albopictus Skuse. PCR was used for molecular identification of An. sundaicus complex, by determination of COI, ITS2, and D3 genes. The target mosquitoes were An. epiroticus, which was the predominant species, accounting for 43.8 % of specimens collected. The biting cycle pattern increased during 6:00-8:00 PM and reached a maximum of 6.6 bites/person/hour by 12:00 PM. The mosquitoes varied in popula- tion density throughout the year. The highest biting rate was 37.6 bites/person/ half night in September and the lowest (10.2 bites/person/half night) in January. Nested PCR and real-time PCR techniques were used to detect the malaria para- site in An. epiroticus adult females. Nine of 926 (0.97%) mosquitoes tested were malaria parasite positive: 6 P. falciparum and 3 P. vivax. The infective mosquitoes were found in the dry and early rainy seasons. The overall annual entomological inoculation rate (EIR) in the village was 76.6. The overall parity rate was 74%. A total of 38 cement tanks were used to characterize the nature of the breeding places of An. epiroticus. An. epiroticus larvae coexisted with Aedes and Culex larvae; the maximum larval density was more than 140 larvae per dip in May. Breeding places included fresh, brackish and salt water, typically with full sunlight and mats of green algae on the water surface. The salinity of the water ranged from 0.5 to 119.4 g/l, with a narrow pH range of 8.2-8.7. Dissolved oxygen was highest in Novem- ber (6.27 mg/l) and lowest in March (3.46 mg/l). The water temperature varied between 24.6 and 32.8ºC. Key words: Anopheles epiroticus, bionomic status, malaria vector, Thailand Correspondence: Chamnarn Apiwathnasorn, Department of Medical Entomology, Faculty of INTRODUCTION Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Bangkok 10400, Thailand. Malaria is transmitted by female Tel: +66 (0) 2351 9100 ext 1575; Fax: +66 (0) anopheline mosquitoes which generally 2643 5582 breed in clean, freshwater habitats. In Vol 41 No. 3 May 2010 541 SOUTHEAST ASIAN J TROP MED PUBLIC HEALTH some coastal areas, malaria is transmitted has a total population of 4,368. Pak Nam by brackish-water breeding mosquitoes. In Village is located in southern Mueang Asia, the most important species are An. Rayong District, Rayong Province, near sundaicus s.l. and An. subpictus (WHO, the coast. The area has fishery and fish 2005). Malaria is a potential fatal disease sauce production. There are many dis- found in some hilly-forested areas and in carded fish sauce cement tanks. Malaria some costal areas of Thailand. In Thailand, was first reported in the area in 2002. A the coastal malaria is found annually and malaria outbreak was reported in 2005 of reported by the Ministry of Public Health 14 cases, and cases have been reported (MOPH, 2008). Unlike along Thailand in- yearly since then. A total of 85 malaria ternational borders where malaria vectors cases have been reported in Pak Nam Vil- have been well-studied, very little is lage during 2002 to 2008 with 61 and 24 of known about the malaria vectors in coastal Thai and non-Thai cases, respectively. areas. The bionomics of An. sundaicus s.l. Three cases (4%) were Plasmodium in Southeast Asia are poorly known due falciparum and 82 (96%) were P. vivax. All to little available data. An. sundaicus s.l. is the falciparum malaria cases were non- considered an efficient malaria vector but Thais. Thai patients presented only with its biologic characteristics and vectorial P. vivax infection. capacity are not clearly understood Adult and larval mosquito collections (Dusfour et al, 2004a). An. sundaicus s.l. has A longitudinal study was conducted been incriminated as a secondary vector during May 2007 - April 2008 in Pak Nam of malaria in Thailand (Gould et al, 1966; Village (Mueang Rayong District, Rayong Harinasuta et al, 1974). Recently, An. Province). Mosquitoes were collected sundaicus species A of Southeast Asia was monthly for 2 consecutive nights from 6:00 formally renamed An. epiroticus (Linton to 12:00 PM by human landing catch et al, 2005). method (HLC) outside 3 houses. Mosquito Insufficient information regarding its larvae were sampled from 38 cement tanks status, bionomics, species complex and its (2.5 x 2.5 x 2.5 m) by the dipping method ability as a malaria vector make vector to study breeding places close to adult col- control in coastal zone of Thailand diffi- lection sites and nearby patient houses. cult. The knowledge of An. epiroticus ob- According to a longitudinal entomologi- tained from this study is important for cal survey, cement tanks containing brack- understanding its potential to transmit ish water in the study area are artificial malaria and should be useful for control- breeding places for An. epiroticus. There are ling malaria vectors in coastal areas of no natural breeding places for An. Thailand. epiroticus in the area. Water quality was estimated using the HACH sension 156 MATERIALS AND METHODS portable multi-parameter instrument (Hach Company, Loveland, CO). Study area Laboratory processing of mosquitoes This study was carried out in Pak Adult mosquitoes were identified by Nam Village, Mueang Rayong District, microscopic examination of morphologi- Rayong Province, Thailand. Rayong Prov- cal characters using established taxonomic ince is located 179 km east of Bangkok. It keys for Thai anophelines (Rattanarithikul 542 Vol 41 No. 3 May 2010 BIONOMIC STATUS OF AN. EPIROTICUS et al, 2006). PCR amplification of 7 3 genes: cytochrome oxidase I 6 (COI), internal transcribed /person/hour 5 spacer 2 (ITS2), and domain-3 4 (D3) of 28S rRNA were per- 3 formed for molecular species An. epiroticus identification (Dusfour et al, 2 2004b; Alam et al, 2006). An. 1 Mean no. of 0 epiroticus female ovaries were 6:00 7:00 8:00 9:00 10:00 11:00 PM dissected to determine parity Time (Detinova, 1962). Malaria infec- Fig 1–Biting of An. epiroticus from 6:00-12:00 PM during tion rates in the An. epiroticus fe- May 2007-April 2008. male adults were detected by dissecting each individual into two parts: head-thorax and abdomen. An. lected using the human landing catch tech- epiroticus DNA was extracted using the nique. Morphological and molecular iden- QIAamp® DNA Mini Kit (QIAGEN, Ger- tification revealed An. epiroticus or An. many) following the manufacturer’s in- sundaicus speies A was the only Anopheles structions with slight modification. The mosquito found in study area. An. DNA was subjected to nested polymerase epiroticus was the most abundant species chain reaction (Nested PCR) (Snounou and (43.8%). Other species collected were Culex Singh, 2002) to detect plasmodium posi- quinquefasciatus (42.6%), Cx. sitiens (8.9%), tive samples, then these were examined Aedes aegypti (4.63%), and Ae. albopictus using real time PCR (Swan et al, 2005). (0.07%). Data analysis Biting cycle of An. epiroticus The human biting rate (HBR) was es- Determination of biting cycle for An. timated as the number of mosquitoes cap- epiroticus was carried out during May tured per person per half night. The par- 2007-April 2008 from 6:00-8:00 PM. Biting ity rate of the famale An. epiroticus mos- activities are shown in Fig 1. An. epiroticus quitoes was calculated using percentages. biting gradually increased during the early The sporozoite rate was the proportion of evening (6:00-8:00 PM) then rapidly in- An. epiroticus mosquitoes found with ma- creased during the last evening (9:00-12:00 laria parasites divided by the number of PM). An. epiroticus biting was unimodal An. epiroticus mosquitoes tested. The risk throughout the year. The peak An. for mosquito-borne infection was esti- epiroticus activity was at midnight (9:00- mated using the entomological inoculation 12:00 PM) with 6.6 mosquitoes per person rate (EIR): the number of bites by an in- per hour. fectious mosquito per person per day Seasonal abundance of An. epiroticus (Macdonald, 1957). An. epiroticus was caught throughout the year (Fig 2). The mean number of An. RESULTS epiroticus collected fluctuated from 10.2- Mosquito abundance 37.6 mosquitoes per person per half night. A total of 3,048 mosquitoes within 5 The highest human biting rate was ob- species belonging to 3 genera were col- served in September, a month with high Vol 41 No. 3 May 2010 543 SOUTHEAST ASIAN J TROP MED PUBLIC HEALTH 40 450 were used to determine 35 400 plasmodium infection by 350 30 nested PCR. Nine (0.97%) 300 head-thorax portions were 25 250 plasmodium positive. Us- An. epiroticus 20 ing real-time PCR determi- 200 15 nation of malaria type in 150 Rainfall (mm) 10 the 9 positive samples, 6 Mean no. of 100 were P. falciparum (66.7%), landings per person half night 5 50 and 3 were P.
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