Isolation of West Nile Virus from Mosquitoes, Crows, and a Cooper's Hawk in Connecticut

Author:

Anderson John F.1,Andreadis Theodore G.2,Vossbrinck Charles R.2,Tirrell Shirley3,Wakem Edward M.4,French Richard A.4,Garmendia Antonio E.4,Van Kruiningen Herbert J.4

Affiliation:

1. Department of Entomology,

2. Department of Soil and Water, the Connecticut Agricultural Experiment Station, Post Office Box 1106, New Haven, CT 06504, USA.

3. Department of Epidemiology and Public Health, Yale University School of Medicine, New Haven, CT 06512, USA.

4. Department of Pathobiology, University of Connecticut, 61 North Eagleville Road, Storrs, CT 06269, USA.

Abstract

West Nile (WN) virus, a mosquito-transmitted virus native to Africa, Asia, and Europe, was isolated from two species of mosquitoes, Culex pipiens and Aedes vexans , and from brain tissues of 28 American crows, Corvus brachyrhynchos , and one Cooper's hawk, Accipiter cooperii , in Connecticut. A portion of the genome of virus isolates from four different hosts was sequenced and analyzed by comparative phylogenetic analysis. Our isolates from Connecticut were similar to one another and most closely related to two WN isolates from Romania (2.8 and 3.6 percent difference). If established in North America, WN virus will likely have severe effects on human health and on the health of populations of birds.

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

Reference28 articles.

1. Asnis D., et al., Morbid. Mortal. Wkly. Rep. 48, 845 (1999).

2. Identification of a Kunjin/West Nile-like flavivirus in brains of patients with New York encephalitis

3. Multiple Isolations of Eastern Equine Encephalitis and Highlands J Viruses from Mosquitoes (Diptera: Culicidae) During a 1996 Epizootic in Southeastern Connecticut

4. . Mosquitoes were captured in dry ice–baited Centers for Disease Control miniature light traps. One mosquito trap was placed at each location per night; the numbers of traps per site ranged from 1 to 6. Mosquitoes were transported alive to the laboratory where they were identified and grouped (pooled) according to species collecting site and date. Numbers of mosquitoes per pool ranged from 1 to 50. Mosquitoes were stored at −80°C until tested for virus. Upon thawing mosquitoes were triturated in tissue grinders or mortars with pestles in 1 to 1.5 ml of phosphate-buffered saline (PBS) containing 0.5% gelatin 30% rabbit serum antibiotic and antimycotic. After centrifugation for 10 min at 520 g 100-μl samples of each pool of mosquitoes were inoculated onto a monolayer of Vero cells growing in a 25-cm 2 flask at 37°C in 5% CO 2 . Cells were examined for cytopathologic effect for up to 7 days after inoculation.

5. C. G. Sibley and B. L. Monroe Jr. Distribution and Taxonomy of Birds of the World (Yale Univ. Press New Haven CT 1990). Common and scientific names of birds are used in accordance with those listed in this book.

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