Journal of the American Control Association, 19(2):151-154,2OO3 Copyright O 2003 by the American Mosquito Control Association, Inc.

BIONOMICS OF EPIDESMUS ASSOCIATED WITH VIRUS IN

P C, KANOJIA

National Institute ofVirology,20-A, Dr. Ambedkar Road, PO Box Il, Pune-4|1001, India

ABSTRACT, Various biological characteristics, including seasonal abundance, diurnal resting habits, feeding behavior, larval habitats, and oviposition, of Culex epidesmus were studied in Gorakhpur District, Uttar Pradesh, India, from 1990 to 1999. Prevalence of adults was recorded between June and October, with a peak in August. The bushy undergrowth of mango and teakwood plantations served as diurnal resting habitat. Blood-meal anal- ysis of wild-caught females showed that the most of them had fed on bovines. Temporary pools of rainwater and pa

KEY WORDS Culex epidesmas, adult population, larval habitats, oviposition, Japanese encephalitis virus, Gorakhpur District

INTRODUCTION ural drainage, flooding and waterlogging occur dur- ing the rainy season in the area. Culex epidesmus (Theobald) belongs to the Cl. Mosquito collections: Weekly visits were made bitaeniorhynclzrzs subgroup of mosquitoes. Several to 3 villages where JE was known to be present species of this group are widely distributed in (Bhathat, Naharpur, and Kusmi) to collect adult southeast and south Asian countries (Reuben 1994), specimens from resting sites such as cattle sheds but C.r. epidesmus is mainly confined to the Indian and in and around pig enclosures with the help of subcontinent, particularly India, , , mouth aspirators and flashlights at dusk. During the and Pakistan. The record from is very day, adults were collected from indoor and outdoor doubtful because it has never been conflrmed by resting places. Immature stages were sampled from subsequent studies (Sirivanakarn 1976). One isolate natural breeding habitats with dippers and pipettes. of Japanese encephalitis virus (JE) has been ob- These specimens were transferred to white enamel tained from this species in the Bankura District of trays containing water and green algae obtained West Bengal, India (Banerjee et al. 1979). from the same breeding sites for rearing to adults. This species has often been mentioned in the lit- Adult specimens were identified with the help of erature because of its association with JE (Burke keys provided by Barraud (1934) and Sirivanakarn and Leake 1988, Reuben et al. 1994). However, the (1976). Field-caught fully engorged females were majority of published work has dealt only with the held in the laboratory to record the egg-laying pat- prevalence of adult populations (Dhanda and Kaul tern. Identification of host blood meals from fully 1980). In view of this, an attempt was made to fed field-caught Cx. epidesmus was carried out by study the bionomics of this species, including adult the double immunodiffusion technique, which is a biology and breeding habitats, during a long-term modifled version of the agarose gel diffusion tech- entomological surveillance of JE in Gorakhpur Dis- nique published by Rao (1984). trict. Uttar Pradesh. India. from l99O to 1999. RESULTS MATERIALS AND METHODS Adult abundance Study area: Gorakhpur District (26'16' to During the present stldy,7,276 female Cx. epi- 27'30'N, 83'6' to 86"56'E) is situated in the north- desmus were collected in 3,871 man hours at dusk. eastern corner of Uttar Pradesh State and lies im- Monthly distribution of adult Cx. epidesmus en- mediately south of the Himalayan slopes. The countered in different years is presented in Table 1. southwest monsoon usually commences in the 2nd Adult populations appeared in June at a very low week of June and lasts to the end of September, density (r : 15; 0.04 per man hour density with maximum precipitation in August. The area IPMDI). Adult populations showed an upward receives total annual rainfall varying from 807 to trend in July (n : 1447;4.0 PMD) and reached a 1,564 mm. Relative humidity ranges from 72 to peak in August (n : 4,327; l3.O PMD). Thereafter, 857o. May is the hottest month of the year, with a population density declined in September (n : maximum temperature of 43"C, whereas January is 1,3191.4.O PMD) and was greatly reduced in Oc- the coolest month of the year, with a minimum tem- tober (n : 164:1.0 PMD). No adult Cx. epidesmus perature down to 4.1"C. Because of inadequate nat- were found from Januarv to Mav and November to 152 JounNx-op rnn AvenrcANMoseurro CoNrnol AssocrarroN Vol.

Table 1. Monthly incidence of female Culex epidesmus collected in different years in Gorakhpur District.r

Jan. Feb. March April May June July Aug. Sept. Oct. Nov. Dec. 1990 l/N 212/3.4 807/15.0 too/2.1 9/0.2 t991 5/0.1 20U3.4 t93/4.0 t39/2.9 3t/0.6 t992 2/N 119/2.4 1,266/26.4 141t2.4 19lo.s r993 l/N 153/3.7 559/17.O 364/7.O 7/0.2 t994 l/N 5/0.4 132/4.0 t08t5.7 24/t.O I 995 2/O.l 233/6.3 t49/4.1 t29/3.8 7lO.4 t996 l/N t34/5.4 244/8.7 t33/4.2 21/O.7 1997 7/O.4 t71/14.3 65/3.O 4/O.2 r998 17/0.7 127/7.5 79/3.0 27/1.5 4/O.2 t999 2/N 366/10.8 679/20.6 6U5.1 l5l1.0 Total l5lN t,447t4.O 4,327/t3.O t,3t9t4.O t64/l.o 4/N rMosquitoes were collected by aspiration at dusk fiom l9l5 to 2015 h. The values given ue total number of mosquitoes/total per man hour density. N, < 0.1.

December except once in 1998 when a few adult season, temporary rainwater pools and paddy fields specimens were collected up to the lst week of No- formed good breeding sites. With the end of the vember. rains, receding water levels exposed aquatic vege- tation, including filamentous green algae. This ad- versely affected breeding of Cx. epidesmus. Diurnal resting habits Fifty-five adult Cx. .epidesmus were collected Oviposition during daytime resting collections made from 1990 to 1994. Of these, 54 specimens were captured Thirty-three fully engorged, fi eld-caught females from bushy undergrowth of mango and teakwood were held in a separate glass jar containing water plantations. However, collections made in other and green algae to record the egg-laying pattern. outdoor vegetation, including sugarcane fields, Of these, 25 females laid egg rafts on algae mats, which are abundantly found in the area, did not whereas 8 females did not lay any eggs. Freshly yield a single specimen. Only I specimen was re- laid egg rafts were found to be boat shaped and covered from inside a house. individual eggs were a white-cream color. The number of eggs deposited by individual Cx. epi- desmus varied widely. Thirteen females deposited Feeding behavior between I 18 and 200 eggs, whereas 12 females de- Fifteen blood smears prepared from gut contents posited from 2O2 to 288 eggs, with an average of of field-caught Cx. epidesmus were tested against 199 eggs per female. human, bovine, and pig antisera. Of these, 12 (807o) were found positive for bovine and only I Discussion (1Va\ reacted with both bovine and human antisera. Two smears did not react with anv antisera. Little information is available on the occurrence of Cx. epidesmus in India. Barraud (1934) reported that this species is rare but widely spread from Pun- Larval habitats jab to Assam. Studies carried out in the Bankura As many as 178 larval Cx. epidesmu,r were sam- District of West Bengal reported occurrence of pled from a variety of breeding places, including adult populations from July to November (Mahadev paddy fields, swamps, perennial ponds, and tem- et al. 1978). The prevalence ofthis species was also porary rainwater pools. Larval populations were lst recorded in Dhanbad District of Bihar State from observed in July and continued to be observed until August to September (Kaul et al. 1982). The pre- October, with the highest numbers in August. No sent study confirms these observations and shows Iarvae were found from November to June except a clearly defined monthly incidence in the adult once in 1998 when a single larva was obtained in population, with lst appearance in June, a peak in November. Distribution of larvae by habitat was as August, and declining populations through Septem- follows: 116 (65.2Vo) in temporary rainwater pools, ber. Adults were rarely collected in November. In 34 (l9.l7o) in paddy fields,22 (l2.4Vo) in swamps, the Gorakhpur region, the monsoon begins in mid- and 6 (3.4Vo)in perennial ponds. Most of the breed- June and remains active until September, with max- ing sites were fully or partially exposed to sunlight imum precipitation in August. During the monsoon and had abundant vegetation, particularly filamen- period, water bodies are formed because of water- tous green algae with some scum on the water sur- logging and these become colonized by aquatic face. These breeding sites usually had clear water plants, particularly filamentous green algae. These and a muddy bottom. At the beginning of the rainy conditions support the breeding of Cx. epidesmus. JuNp 2003 BroNorr.trcsoF Cx. EptDESMUsAND Irs RpuuoN To JE VIRUS 153

The high population density in August may be ex- quitoes. However, the eggs, when freshly deposited, plained by the availability of excessive breeding were reported to show a light green coloration. places, and the decline in populations through Sep- During the present study, C-{. epidesmus was ob- tember can be attributed to the loss of eggs and served to lay eggs in boat-shaped rafts and the immature stages because of water overflowing from number of eggs laid by individual mosquitoes var- the breeding places. ied from ll8 to 288, with a mean of 199. In earlier studies, diurnal resting sites of Cx. ep- In Gorakhpur, prevalence of adult mosquitoes idesmus were not observed. But during the present showed I peak exclusively toward the beginning of study, Cx. epidesmus were found resting in bushy the JE season, that is, in August. The studies carried undergrowth of mango and teakwood plantations. out in Bankura District of West Bengal (JE endemic Collections made in open sugarcane fields, which area) have also shown a high density of adult pop- included other vegetation, did not yield a single ulations during August and adults were collected specimen in spite of the efforts made, suggesting on pig, chicken, cattle, and human baits (Mahadev that Cx. epidesmus prefers more covered ground et al. 1978). In addition to this, Cx. epidesmus has vegetation as diurnal harborages rather than ex- been found to be naturally infected with JE (Ba- posed a"reas. nerjee et al. 1979), which indicates that this species Information available on the feeding habits of has access to viremic hosts. Hence, it seems that Cx. epidesmus is scanty. Blood-meal analyses of Cx. epidesmus may be playing some role in the Cx. epidesmus collected from Sundargargh District, natural history of JE. Further experimental trans- Orissa, were found positive for bovines (Dash et al. mission studies are required to know its vectorial 20Ol). The present study confirmed that Cx. epi- potential. desmus predominantly feeds on bovine hosts. In contrast, 2 closely associated species, Culex bitae- ACKNOWLEDGMENTS niorhynchus Giles and Culex infula Theobald, have wide host ranges. Culex bitaeniorhynchus is con- I wish to thank D. A. Gadkari, former Director, sidered to be aviphilic, although these mosquitoes and V. S. Padbidri, officer-in-charge, National In- occasionally are found positive for cattle, pigs. stitute of Virology (NIV), Pune, for their support dogs, and humans (Christopher and Ruben 1971, and encouragement. The assistance rendered by the Reuben l97l). Culex infula feeds mainly on cattle field staff of NIV Field Station, Gorakhpur, is but also shows a low proportion of human and bird gratefully acknowledged. I also wish to thank the blood meals (Reuben et al. 1992). Director of the Meteorological Centre, Lucknow, B. The breeding habitats of Cx. epidesmus were not P Srivastava, who furnished meteorological data well known. In earlier work, larvae were collected for Gorakhpur District. from uncultivated rice fields in Sri Lanka, but sub- sequent visits made to the same localities failed to REFERENCES CITED recover any specimens (Sirivanakarn 1976). But in the present study, breeding habitats of Cx. epides- BanerjeeK, Mahadev PVM, Ilkal MA, Mishra AC, Dhan- Modi G, Kaul HN, Shetty PS, mus were identified. This species was found breed- da V GB, Geevarghese George PJ. 1979. Isolation of Japaneseencephalitis vi- ing in temporary rainwater pools, paddy fields, rus from mosquitoescollected in Bankura District, West swamps, and perennial ponds containing with green Bengal, during October 1974 to December 1975.Indian algae. This suggests that females prefer to lay eggs J Med Res69:201-205. in water containing green algae and larvae depend Barraud PI. 1934. 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