Mosquito Collections Following Local Transmission of Pi^A,S M Od I Um Falc I Paru M Malaria in Westmoreland County, Virginia
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Jourtal of the Ameican Mosquito Control Association, 16(3):219-222,2O00 Copyright @ 2000 by the American Mosquito Control Association, Inc. MOSQUITO COLLECTIONS FOLLOWING LOCAL TRANSMISSION OF PI^A,S M OD I UM FALC I PARU M MALARIA IN WESTMORELAND COUNTY, VIRGINIA DANIEL STRICKMAN,' THOMAS GAFFIGAN,' ROBERT A. WIRTZ,' MARK Q. BENEDICT' CRISTINA SALAZAR IU{IrI]ERTY.2 RACHEL S. BARWICK'AND HOLLY A. WILLIAMS' ABSTRACT. A 63-year-old woman from Colonial Beach, Westmoreland County, VA, was diagnosed with Pl.asmodiumfalciparum malaria on July 19, 1998. The woman had no history of international travel, intravenous drug use, blood transfusion, or other risk factor for contracting the disease. She seldom left the county and generally spent her evenings indoors, leading to the conclusion that she had been bitten locally by an infected mosquito. Colonial Beach is host to a population of migrant agricultural laborers from areas in which malaria occurs, but a blood survey of 89 Haitians and Mexicans failed to find Plastnodiwn parasites, specific antibodies, or clinical cases of malaria. Mosquito surveys were conducted during 2 days (July 22 and 28, 1998) with carbon- dioxide-baited light traps, larval and pupal collections, and landing collections. Thirteen species of mosquitoes were identified morphologically, including 4 potential vectors: Anopheles crucians, An. punctipennis, An. smar- agdinus (new state record), and An. quadrimaculatzs s.s. (new state record). Identifications of the latter 2 species were confirmed by sequencing ofthe ITS2 DNA region from adults reared from locally collected larvae. Anoph- eles smaragdinzs was the most cornmon biting species among the potential vectors, although An. crucians was the most abundant in other kinds of collections. In addition, Ae. albopictus was collected in Westmoreland County for the lst time. KEY WORDS Plasmodium falciparum, malaria, Virginia, Anopheles quadrimaculatus, Anopheles smaragdi' nus, Aedes albopicus INTRODUCTION for cooling. Close examination of the screens showed that they had adequately fine mesh (22llin- On July 17, 1998, a 63-year-oldwoman living in ear inch) and were in generally good repair. The Colonial Beach, Westmoreland County, VA, be- only imperfections were small gaps between the came ill with fever, myalgia, a stilf neck, and di- screening frames and the sills and small tears in a arrhea. She was admitted to a local hospital on July few of the screens. 19 and promptly diagnosed and treated for malaria The neighborhood consisted of large lots (ap- causedby Plasmodiumfalciparum (Welch). The di- proximately I ha) arranged linearly along a 2-lane agnosis was confirrned from blood smears exam- road. East of the lots and separating some of them ined by the Biology and Diagnostics Branch, Di- were dense stands of trees, including tulip poplar vision of Parasitic Diseases, National Center for (Liriodendron tulipiftra L.), white oak (Quercus Infectious Diseases.Because the woman had no re- alba L.), sycamore (Platanus occidentalis L.), tu- cent history of travel outside the local area, no pelo (Nyssa sylvatica Marsh), bench (Fagus gran- blood transfusion, no intravenous drug use, and no difuIia Ehrh.), and easternred cedar (luniperus vir- previous malaria, her infection must have been ac- giniana L.). A slow-flowing stream in the woods quired locally from the bite of a mosquito. No other formed an extensive area of small ponds and c,rses of malaria were detected in the county, de- swamps draining into a small, tree-lined stream spite extensive efforts to find them. The most likely (Monroe Creek). Other aquatic habitats within 2 km source for the original introduction of P. falciparum of the house included a nearly undisturbed 2-acre was from among the large pool of migrant agricul- pond (locally called Betty Lake) in the forest and tural workers living in Colonial Beach, although in- a pond with few trees around it located adjacent to people terviews and blood samples of 89 from Monroe Campground. No organized mosquito (4) Mexico (85) and Haiti taken July 20-25,1998, abatement activity exists in the county and the state failed to find any evidence of infection. does not have central mosquito control assetsto be The patient lived in a 2-story, modern, screened used in case of an emergency. home, which was in excellent condition. She re- ported that she rarely went outside in the evening, except sometimesbriefly to walk her dog. Signifi- MATERIALS AND METHODS cantly, the home was not air conditioned at night, Three different kinds of mosquito collections relying on open, screenedwindows and ceiling fans were made between July 22 and 28, as follows: landing collections by 2 people on 2 nights from 2fiD to 22OOh, collections in portable light traps I Department of Entomology, Walter Reed Army Insti- tute of Research,Washington, DC 20307-5100, (American Biophysics, Jamestown, RI) supple- 2 Entomology and Epidemiology Branches,Division of mented with carbon dioxide from dry ice operated Parasitic Diseases,Centers for DiseaseControl and Pre- on 2 nights, and collections of larvae from 12 dif- vention, Atlanta, GA 30341-3724. ferent sites. Light trap and landing collections were 220 JounNn- oF THE AMERTcIN Moseurro Covrnol AssocnnoN Vol. 16, No. 3 made in the backyard of the patient's home and Table l. Mosquito species collected JuLy 23-28, 1998, larval collections were all made within 2 km of the in the vicinity of a locally transmitted case of home. These larvae were reared in the laboratory, Plasmodium falciparum malaria (date of onset July 17) in Westmoreland VA. preserving exuviae and adults for morphologic ex- Colonial Beach. County, amination. In addition, anopheline larvae were col- Number lected on August 31, reared, and the resulting adults collected from the DNA Quadrimaculatus Group retained for Larvae/ Land- sequence analysis of the ribosomal RNA intergenic Species pupael Trap2 ing' spacer 2 (ITS2) region using a method similar to that of Cornel et af. (1996). Individual sequences Aedes albopictus (Skuse) 0 03 Ae. canadensis (Theobald) 0 and primers used for polymerase chain reaction am- 03 Ae. triseriatus (Say) 0 117 plification and sequencing are available from the Ae. vexans (Meigen) 0 12 authors (M.Q.B. or C.S.R.). The DNA sequences Anophele s crucians Wiedemann t2 11 I from the Colonial Beach collection were compared An. punctipennis (Say) 0 20 with other Anopheles ITS2 by alignment and clus- An. quadrimaculatus Say 7 0l tering using the program PILEIIP (Wisconsin Pack- An. smaragdinas Reinert r0 15 age Version 9.1, Genetics Computer Group, Madi- Coquillettidia perturbans son, WI) with gap creation and extension penalties (Walker) 0 o2 of 2 and 5, respectively. Culex erraticus (Dyar and Knab) 20 l1 43 Morphologic identifications of culicine species Cx. salinarius Coquillett 0 623 and anopheline species groups were made accord- Psorophora ferox ing to Darsie and Ward (1981). The report by Re- (Von Humboldt) 0 01 inert et al. (1997) was used to identify members of U ranotae nia sapp hi rina the Quadrimaculatus Group and the work of Floore (Osten Sacken) ll 10 et al. (1976) was used for the Crucians Group. tFrom 12 collections made at the mugins of ponds, swamps, When associated immature forms were not avail- and streams. able (light trap and landing collections), adults of 'Ponable light trap supplemented with crbon dioxide from dry the Quadrimaculatus Group were identified with ice, operated 2 nights. 3 people confidence by comparison to adults from reared Two collecting from 2000 to 22OO h for 2 nights. specimens. Because adults of species of the Cru- cians Group are indistinguishable, adults from traps and landing collections were assumed to be the tered closely with An. quadrimaculatus s.s. How- sarne species as the reared specimens. All speci- eveB a more diverse group clustered with Az. mens were deposited at the U.S. National Museum smaragdinus, These results were concordant with as Walter Reed Biosystematics Unit (WRBU) Ac- the morphologic identifi cations. cession 1683. DISCUSSION RESULTS Any decision on the identity of the vector in Co- The results of the collections (Table l) reflected lonial Beach must be consideredtentative, because the rich variety and abundance of mosquitoes at the small number of anophelines collected preclud- Colonial Beach. The three most abundant biting ed direct examination for sporozoites.The 2 least mosquitoes v,tere Culex erraticus (Dyar and Knab), likely species were Anopheles punctipennis (Say) Cx. salinarius Coq., and Aedes triseriatus (Say), and An. crucians. Doubts about the importance of but 8 other species were collected in just 2 eve- An. punctipennis as a malaria vector date back to nings. Although Anopheles smaragdinas Reinert the early days of malariology (Smith 1914), af- was the predominant biting anopheline, An. cru- though the species has been proven capable of cians Wied. was most abundant in larval collections tf,ansmitting P. falciparum for up to 92 days and light traps. An interesting aspect of the collec- (Mayne L922). Morc recently, An. punctipennis in tions was the occurrence of anophelines in every California was implicated as a powerful vector of larval habitat examined. The collection of Anoph- Plasmodiumvivax (Grassi and Feletti), basedon its eles quadrimaculatus Say s.s. and An. smaragdinus longevity and short gonotrophic cycle (Jensenet al. were new state records for Virginia and the collec- 1998). The scarcity of An. punctipennis at Colonial tion of Aedes albopictus (Skuse) was a new county Beach, which is apparently typical for this species record (Chester G. Moore, personal cornmunica- in coastal areasof the region (Vogt 1947, Gladney tion). and Turner 1969), makes it an unlikely candidate The DNA sequence analysis was compared to dl as the local vector. Although ample records exist of Anopheles (ITS2) sequences available to the au- naturally infected An. crucians (Floore et al. 1976), thors, including unpublished sequences (C. Porter, this specieswas also an unlikely vector at Colonial personal communication).