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Susceptibility of Carrion Crows to Experimental Infection with Lineage 1 and 2 West Nile Viruses - Volume 21, Number 8—August 2015 - Emerging Infectious Disease journal - CDC

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Susceptibility of Carrion Crows to Experimental Infection with Lineage 1 and 2 West Nile Viruses - Volume 21, Number 8—August 2015 - Emerging Infectious Disease journal - CDC

Volume 21, Number 8—August 2015


Susceptibility of Carrion Crows to Experimental Infection with Lineage 1 and 2 West Nile Viruses

Stephanie M. Lim, Aaron C. Brault, Geert van Amerongen, Angela M. Bosco-Lauth, Hannah Romo, Varsha D. Sewbalaksing, Richard A. Bowen, Albert D.M.E. Osterhaus, Penelope Koraka1, and Byron E.E. Martina1Comments to Author 
Author affiliations: Erasmus Medical Center, Rotterdam, the Netherlands (S.M. Lim, G. van Amerongen, V.D. Sewbalaksing, A.D.M.E. Osterhaus, P. Koraka, B.E.E. Martina)Centers for Disease Control and Prevention, Fort Collins, Colorado, USA (A.C. Brault, A.M. Bosco-Lauth, H. Romo)Colorado State University, Fort Collins (R.A. Bowen)Artemis One Health Research Institute, Utrecht, the Netherlands (A.D.M.E. Osterhaus, P. Koraka, B.E.E. Martina)


West Nile virus (WNV) outbreaks in North America have been characterized by substantial die-offs of American crows (Corvus brachyrhynchos). In contrast, a low incidence of bird deaths has been observed during WNV epidemic activity in Europe. To examine the susceptibility of the western European counterpart of American crows, we inoculated carrion crows (Corvus corone) with WNV strains isolated in Greece (Gr-10), Italy (FIN and Ita09), and Hungary (578/10) and with the highly virulent North American genotype strain (NY99). We also inoculated American crows with a selection of these strains to examine the strains’ virulence in a highly susceptible bird species. Infection with all strains, except WNV FIN, resulted in high rates of death and high-level viremia in both bird species and virus dissemination to several organs. These results suggest that carrion crows are highly susceptible to WNV and may potentially be useful as part of dead bird surveillance for early warning of WNV activity in Europe.
West Nile virus (WNV), a flavivirus (family Flaviviridae) transmitted by mosquitoes, uses birds as its primary vertebrate reservoir host. WNV has an extensive geographic range that includes Europe, Africa, the Middle East, southern Asia, and Australia (1). In 1999, WNV emerged in North America, where it was first detected in New York, New York. The virus subsequently spread rapidly across the continent, becoming the leading cause of arboviral encephalitis in humans and horses (2), and it was associated with deaths among at least 326 bird species (3). High death rates are most frequently observed among passeriform birds, of which the familyCorvidae comprises the most highly susceptible species to WNV (4). In particular, deaths among the American crow (Corvus brachyrhynchos) have been used to track the spread of the virus across many parts of North America (58).
Since 2008, WNV has been responsible for outbreaks throughout central and southeastern Europe, affecting countries such as Greece, Italy, Hungary, Romania, and Croatia and constituting a serious veterinary and public health problem. Fatalities have been reported among wild birds in Europe, such as eagles (9,10), sparrow hawks, goshawks, geese, and falcons (1113). However, death rates among birds in Europe have been low, and no clustered death events have occurred, even when cases were associated with outbreaks of severe human and equine WNV infections (1417). Several theories have been proposed to explain the low death rates among birds in Europe: limited or insufficient monitoring of deaths among wild birds in Europe; development of immunity among birds from infections acquired on wintering grounds (18); and circulation of WNV strains in Europe with reduced virulence for birds.
Experimental infection of American crows with the North American genotype of WNV (NY99) has shown that the strain has a highly pathogenic phenotype: viremia titers exceeded 9 log10 PFU/mL, and all infected birds died (1923). However, the lack of WNV-associated bird deaths in Europe suggests that European birds might not be susceptible to WNV or that WNV strains from Europe are not virulent to birds. Thus, we evaluated the susceptibility of the European equivalent of the American crow, carrion crows (Corvus corone), which are ubiquitously present across Europe, by injecting them with selected strains of WNV circulating in Europe and with the prototypic NY99 strain. In addition, we inoculated American crows with a selection of these viruses to assess and compare the virulence of WNV strains from Europe in a bird species known to be highly susceptible to WNV. Susceptibility was assessed in terms of death, survival time, magnitude and duration of viremia, and spread of virus to different organs.
Ms. Lim is a graduate student at the department of Viroscience of the Erasmus Medical Centre. Her research interests include the pathogenesis of (neurotropic) arboviruses.


We thank Vittorio Sambri, Luisa Barzón, Giorgio Palù, and Tamás Bakonyi for providing the low-passage isolates used in this study. We also thank Tanja Schouten and Angela Gomersbach for their excellent technical assistance; Jeroen Roose and Peter van Run for their technical assistance with the immunohistochemistry; and Thijs Kuiken for his assistance with the analysis of the histological stainings.
The research leading to these results has received complete funding from the European Community's Seventh Framework Programme (FP7/2007-2013) under the project “VECTORIE,” European Commission grant agreement number 261466. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Trapping of carrion crows was performed with the assistance of the Gemeentewerken Rotterdam under permission obtained from the Ministry of Agriculture (registered under no. FF/75A/2011/031). Experimental inoculations were performed under protocol number 122-12-12 with permission obtained from the Animal Ethics Committee of Erasmus Medical Centre. All efforts were made to minimize animal suffering. Trapping of American crows was performed with the assistance of Todd Felix under US Fish and Wildlife Scientific Collecting Permit number MB-91672A. Experimental inoculations of crows were performed under Colorado State University IACUC approval number 10-2078A.


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Technical Appendix

Suggested citation for this article: Lim SM, Brault AC, van Amerongen G, Bosco-Lauth AM, Romo H, Sewbalaksing VD, et al. Susceptibility of carrion crows to experimental infection with lineage 1 and 2 West Nile viruses. Emerg Infect Dis. 2015 Aug [date cited].
DOI: 10.3201/eid2108.140714
1These authors contributed equally to this article.

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