ANOPHELES BARBIROSTRIS/CAMPESTRIS AS A PROBABLE VECTOR OF MALARIA IN , PROVINCE

Dakorn Limrat', Boonchai Rojruthai', Chamnarn Apiwathnasorn2, Yudthana Samung2 and Samrerng Prommongko12

'Office of Vector Borne Disease Control Region 5, Department of Communicable Disease Control, Ministry of Public Health, Nonthaburi; 2Department of Medical Entomology, Faculty of Tropical Medicine, Mahidol University, ,

Abstract. As a result of dramatic increase in malaria cases in from 666 cases in 1995 to 4,381 in 1997, a brief entomological study was carried out during January 1998 to December,1999 in Pa Rai subdistrict where most malaria cases were reported. Of fourteen species of mosquitos found, only Anopheles barbirostris group was the most abundant species throughout the year. Adult identification was not able to confirm species within An. barbirostris group, particularly between An. barbirostris and An. campestris because of morphological resemblance. Therefore, the barbirostris group captured in this study is reported to be either An. barbirostris or An. campesrris. 'The seasonal prevalence of barbirostris/campestris was bimodal in distribution (September and November) and coincided well with malaria occurrence in this area. Human landing collections revealed high adult densities with 20 biteslperson-night for mean indoor density and 53.5 biteslperson-night for mean outdoor density. The biting peak was during 21.00- 24.00 hours. Among 223 barbirostris/campestris dissected for oocysts and sporozoites only one gut from the outdoor collection in November was infected with oocysts. There were no sporozoites detected in salivary glands of all mosquitos collected. 'This appears to indicate that in the absence of major vectors local species may serve as potential transmitters of malaria in Thailand.

INTRODUCTION confined to forested rural areas principally along the borders with Myanmar and Cambo- Malaria has been recognized as a major dia, an epidemiology characterized by multi- disease problem in Thailand since 1918 with drug resistant Plasmodium falciparum, an estimate of 40,000 to 50,000 deaths annu- exophagic and exophilic behavior of the major ally. Malaria was the leading cause of death anopheline vectors (Anopheles dirus and An. (27 1.7- 194.9 per 100,000) until 1950 (Division minimus) and intensive population movement. of Malaria Centre, 1973). The establishment However, there has been a resurgence of malaria of the National Malaria Control Program with in some areas of the country where malaria a variety of activities including expansion of incidence was previously at a low level, at- primary health care delivery and malaria clin- tributable to population migration due to eco- ics significantly reduced the mortality rate from nomic, demographic and ecological factors 351 per 100,000 population in 1947 to 1.26 (Vijay kadga, 1999). In 1998 malaria transmis- per 100,000 population in 1998 (Vijaykadga, sion occurred in several provinces of southern 1999). At present malaria transmission is largely Thailand; Chumphon, , Phuket but particularly in Surat Thani where malaria cases Correspondence: Chamnarn Apiwathnasorn, De- were highest (17,302 cases) - an increase of partment of Midical Entomology, Faculty of Tropi- 300% the previous year. cal ~~di~i~~,42016 ~~,~i~hi~~~d, ~~~~k~k 10400, these, 91.2% were associated with agricultural Thailand. activity. Thailand's economic crisis resulted in Tel: (+66) 22469000-12, 22460056; Fax: (+66) a movement of people from urban centers to 22468340; E-mail: tmcaw @mahidol.ac.th rural areas to exploit forests for agriculture

Vo1 32 No. 4 December 2001 739 purposes areas where malaria vectors were 81.68% of the total area was devoted cultiva- abundant. In Surat Thani, over 1,600 acres tion of rice (49.5%), maize (33.39%), cassava were exploited. Indigenous as well as imported (31.2%), sugarcane (9.6%) and other cash crops. cases were reported within the country. Field investigations Malaria transmission in Sa Kaeo Prov- ince, an eastern boundary province with Cam- A longitudinal study was conducted from bodia was also atypical. Malaria cases increased January 1998 to December, 1999 in Pa Rai from 666 cases in 1995 to 1,066 cases in 1996 subdistrict (, Sa Kaeo Prov- and 4,381 in 1997; 88.5% of cases were due ince). Mosquito collections to assess biting P vivax. Infection was thought to be intro- activity were made monthly by indoor and duced by migrants. The major vectors, An. outdoor landing captures in three collection dirus, An. minimus and An. maculatus s.1. were stations from 18.00-05.00 hours for two nights. found occasionally. This study presents the Although cattle were scarcely found in the entomological data obtained from Pa Rai, a study area, however, a single cow-baited col- subdistrict of Aranyaprathet, Sa Kaeo Prov- lection was made incidentally in October of ince. The objectives of the study was to 1999 when the mosquitos were most abundant. demonstrate potential of the An. barbirostris The collected mosquitos were identified and group in malaria transmission. examined for parity based on tracheation of the ovaries of mosquitos (Detinova, 1962). Mid- guts and salivary glands were examined by MATERIALS AND METHODS dissection for the presence of oocysts and sporozoites, respectively. Larval surveys were Study area carried out monthly by dipping from all avail- Sa Kaeo is a newly established province able water collections to determine presence upgraded from Sa Kaeo district of Prachin Buri of anopheline mosquitos. Immatures stages were Province in 1993. The province is located 236 brought to the insectary and raised to obtain km from Bangkok. It is administratively di- pupal skins for identification of members of vided into 6 districts: Muang, Watthana Nakhon, the barbirostris group. Blood samples were Aranyaprathet, Ta Phraya, Wang Nam Yen, and examined monthly for malarial parasites by Khlong Hat. According to the annual report of optical microsopy. Climatological data were Malaria Division, Sa Kaeo was regarded as a provided by the Meteorological Department, low-risk area until 1996. Henceforth malaria Ministry of Communications, and were re- transmission in Sa Kaeo was among in the ported as mean monthly temperatures and total highest ten in the country. The highest trans- rainfall. mission (1,498 cases in 1998) of the country occurred in Aranyaprathet on the Thai-Cambo- RESULTS AND DISCUSSION dian border, 70 km away from Sa Kaeo town. This border town is a marketplace where Malaria situation in Aranyaprathet products from are on sale at Ban Khlong Luk, a village 6 krn away. Aranyaprathet Malaria situation in Aranyaprathet was has a total population of 8 1,835 with a popu- less than 600 cases annually until 1996. Between lation density of 75.60 persons/km2, the highest 1997 and 1999, there was a dramatic increase in the province. Three mosquito collection of malaria with 1,496-1,956 cases reported of stations were located in Pa Rai, a subdistrict which more than 95% were due to P vivax with the highest number of malaria cases in and the rest, to P falciparum (Fig 1). Fig 2 Aranyaprathet (281 cases in 1998). Pa Rai demonstrates malaria transmission was appar- covers an area of 31 km2 with a population ently unimodal starting in July, reaching a of 3,899 in 1999 distributed among 8 villages peak in December and declining after Decem- and 847 households. In terms of land use, ber to June.

Vo1 32 No. 4 December 2001 A total of 1,990 anopheline mos- quito~belonging to 6 species were captured by human landing catches. The most abundant species (66.2%) was An. barbirostris/campestris. An. aconitus, another suspected vector was also caught in relatively smaller numbers (9.770 of total collections). Other Anopheles spe- cies collected were An. hyrcanus group 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 (An. peditaeniatus and An. nigerrimus) Year (13.6%), An. nivipes (5.1%), An. Fig I-Malaria cases in Aranyaprathet during 1990- 1999. tessellatus (4.4%), and An. vagus (I .0%). The major vectors, An. dirus, An. minimus and An. maculatus s.1. were not found during the study period. Based on the annual reports of Malaria Division. these main vectors have occurred in low numbers and have even been absent from the study sites since 1980. In contrast to the cow-baited col- lection, An. barbirostris/campestris occurred in small numbers. A total of 1,108 collected mosquitos was comprised 01 ...... l JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC of 65.3% An. peditaeniatus, 21.5% Cx. Month tritaeniorhynchus, 7.170 An. barbirostrid Fig 2-Seasonal variation of slide positive rates in Aranyaprathet. campestris and 1.5% An. aconitus. The data revealed that adult barbirostrid campestris at least from this area were Mosquito abundance anthropophilic. Fourteen species of mosquitos were col- lected by means of human landing catches and Infection rate a cow-baited trap. Identification of female adults All anopheline mosquitos were dissected of members of the barbirostris group was for oocysts and sporozoites. Only An. inaccurate, particularly between An. barbirostris barbirostris/campestris was found infected by and An. campestris because of their similar and malarial parasites. Out of 223 dissected guts overlapping characters. Precise identification of An. barbirostris/campestris from the out- of this species group is based on the exami- door collection in November, only one gut was nation of the pupal skins (Harrison and Scanlon, found infected with 67 oocysts (0.45% infec- 1975). In this study the designation An. tion rate). There were no sporozoites detected barbirostris/campestris is meant to represent in salivary glands of the mosquitos collected. either the species barbirostris or campestris. The species composition of the mosquitos found To date member species of An. barbirostris feeding on human and cattles was consisted group has not been incriminated as a vector of Aedes mediolineatus, Ae. vexans, Anopheles of malaria in Thailand, however, barbirostris aconitus, An. barbirostris/campestris, An. was reported several times to be infected by nigerrimus, An. nivipes, An. peditaeniatus, An. malarial oocysts and sporozoites. Gingrich et tessellatus, Culex bitaeniorhynchus, Cx. gelidus, a1 (1986) determined potential Anopheles spp Cx. quinquefasciatus, Cx. tritaeniorhynchus, vectors throughout Thailand by ELISA sporo- and Mansonia uniformis. zoite analyses and ranked An. barbirostris as

Vol 32 No. 4 December 2001 the second highest positive rates for l? l5 falciparum and l?vivax following An. -$ 12 dirus. Griffith (1955) reported malarial 2 YI L oocysts from a single barbirostris from .-25g 2 9 Chiang Mai Province. In addition the p~ presence of barbirostris has been widely i6 acknowledged by our field workers because its actual distribution is com- mon throughout the country in contrast o to those of the other members of Time barbirostris group. In view of biologi- cal aspect, however, there is a little Fig 3-Biting activity of An. barbirostris/campestriscollected potential for barbirostris being a ma- indoors and outdoors during October-November, 1999. laria vector because the adult barbirostris 80 25 are generally zoophilic in Thailand ,o (Harrison and Scanlon, 1975). The iden- E 20 .$60 tity of An. barbirostris in these studies U) is inconclusive and needs confirmation. 50 15 { 0 .-m The possibility of campestris being a i40 L a 10 m vector of malaria is more plausible than g 30 2 that of barbirostris as regarded by Reid 1 20 5 (1968) that An. campestris is highly lo anthropophilic and ranked as the third o o species of Anopheles in Southeast Asia JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC Month most attracted to man. Owing to the unknown distribution and feeding hab- Fig 4-Indigenous cases and An. barbirostris/cam- its of Thai campestris coupled with the pestris density in Pa Rai Subsistrict, Aranyaprathet (1998). problem of adult identification, its vec- tor status for malaria in Thailand is campestris from human-baited collections during problematical at present. 19.00-24.00 hours in Pathum Thani Province and by Moorhouse and Wharton (1965) which Parity rate was between 20.00-02.00 hours in Malaysia. Among 941 An. barbirostris~campestris More mosquitos were collected from outdoor dissected for parity based on the coiling of catches. The highest density of 12 bitestper- ovarian tracheoles, the parous rate was mod- son-hour occurred at 22.00 hours and 4 bitest erately high ie, 60.7%. person-hour at 23.00 hours for outdoor and indoor catches, respectively. In comparison to Bionomics of An. barbirostris/campestris the major malaria vectors, An. dirus, An. Biting cycle: Determination of biting cycle of maculatus s.1. and An. minimus, reported in An. barbirostris/campestris was carried out various areas of Thailand, An. barbirostris/ during October to November when population campestris were active in higher densities (20 densities were highest. Biting activities were bitestperson-night for mean indoor density and between 19~00-05,00hours as shown in Fig 3. 53.5 bitestperson-night for mean outdoor den- The biting patterns similarly fluctuated through- sity). out the night from 21.00-05.00 hours with the highest peaks between 22.00-23.00 hours and Seasonal prevalence smaller peaks after 01.00 hour. The biting ac- The seasonal abundance of An. barbirostrid tivity was similar to that described by Gould campestris was determined by night observa- et (1965) which found biting peak of An. tions with human landing catches. The sea-

742 Vo1 32 No. 4 December 2001 sonal trend showed a bimodal distribution with An. campestris by associated pupal skins. An. peak biting rates of 19 and 15 bitestperson- campestris and An. hyrcanus group were con- night in September and November, respec- sistently detected in the same sites throughout tively for indoor populations and 36.5 and 78 the year. bitestperson-night in June and September, respectively for outdoor populations (Fig 4). SUMMARY The mosquito abundance synchronised with malaria occurrence in this area. The outdoor An. barbirostris/campestris is a probable density of An. barbirostris/campestris from May malaria vector based upon its density, biting to June was possibly responsible for an initial habits, seasonal population dynamics, the find- rising in malaria cases from June to July ing of an infected mosquito and the absence followed by a big peak of malaria cases from of other main vectors of malaria. Unpublished July to February which was synchronous with works detected An. barbirostris/campestris indoor and outdoor peaks of An. barbirostris/ positive with sporozoites by either ELISA or campestris abundance from July to December. dissecting technique, however, inaccurate iden- tification of the adults made it impossible to Malaria transmission in the study area incriminate any of these species as an addi- appears to be seasonal possibly associated with tional malaria vector of Thailand. Although sugarcane cultivation which covered an area only one barbirostris/campestris was found of 18,190 rai (29.1 km2). The sugarcane season infected with oocysts in this investigation, the was during May-February. Cultivation started findings seem to indicate that malaria in Pa during May to July and was harvested during Rai may be transmitted by its local mosquito January to February. It was estimated that 1 species (An. barbirostris/campestris) in the persontraitday was needed for planting and 5 absence of the main vectors. Further investi- personstraitday for cutting. This labor force gation on the species confirmation and bion- was mostly immigrants from Cambodia. Ac- omics of An. barbirostris/campestris are needed cording to an immigration officer in Aranya- to clarify its vector status. prathet, some 3,000 Cambodians cross daily via Wattana Nakhon district. Reports showed that 6.7% of 2,669 immigrants examined for ACKNOWLEDGEMENTS malaria parasites were found infected; of these 165 (6.2%) blood smears were positive We wish to thank Professor David Moly- for /? vivax. It was observed that many im- neux, Liverpool School of Tropical Medicine migrants working for sugarcane cultivation slept for critical reviewing the manuscript. This study overnight in the temporary huts in sugarcane is a collaboration of the Division of Malaria fields with no means of protection from Control, Department of Communicable Disease mosquito bites. This constituted a suitable source Control, Ministry of Public Health, Thailand of malaria infection in the study area. and the Faculty of Tropical Medicine, Mahidol University. Larval surveys Six species of Anopheles including aconi- REFERENCES tus, campestris, hyrcanus group, nivipes and vagus were collected commonly on the periph- Detinova TS. Age-grouping methods in Diptera of ery of villages in semi-permanent pools, ponds medical importance. With special reference to and rice fields and in pits and marshes within some vectors of malaria. Geneva: World Health villages. Most contained water from 5-30 cm Organization, WHO Monogr Ser 1962; 47. deep, algae, low grasses and other aquatic plants Division of Malaria Control. Annual Report of Ma- common around the periphery with partial shade. laria, 1997. Division of Malaria Control, Depart- All immatures of barbirostris group collected ment of Communicable Disease Control, Minis- in these breeding habitats were identified as try of Health, Thailand. 1997.

Vol 32 No. 4 December 2001 Gingrich JB, Wirtz RA, Pilakasiri C, et al. ELlSA (Diptera:Culicidae). Contrib Am Ent lnst 1975; sporozoite evaluation of potential Anopheles spp 12: 1-307. vectors collected throughout Thailand. Pattaya Ministry of Public Health. Public health in Thailand. City: Second Conference on Malaria Research, 8'h ed. 1973: 1-124. Thailand. 2-4 December, 1986: 176. Moorhouse DE, Wharton RH. Studies on Malayan Gould DJ, Esah S. Pranith U. Relation of Anopheles vectors of malaria; method of trapping and ob- aconitrcs to malaria transmission in the central servations on biting cycles. J Med Entomol 1965: plain of Thailand. Trans R Soc Trop Med Hyg I: 359-70. 1965; 59: 441 -2. Reid JA. The Anopheles barbirostris group (Diptera: Griffith ME. A note on An. minimus Theobald as a Culicidae). Bull Entomol Res 1962; 3: 1-57. malaria vector in Thailand. Rome: VI Interna- Reid JA. Anopheline mosquitos of Malaya and Bor- tional Congreas of Microbiology, 1955; 5: 565- neo. S~udlnst Med Res Malaya 1 968; 3 1 : 1-520. 7. Vijaykadga S. Malaria transmission in Thailand. In Harrison BA, Scanlon JE. Medical entomology stud- Malariology BE 2543. Bangkok. 1999: 36-48. ies - 11. The subgenus Anopheles in Thailand (In Thai).

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