Nicholas Komar

Experimental Infection of Horses with West Nile virus

A total of 12 horses of different breeds and ages were infected with West Nile virus (WNV) via the bites of infected Aedes albopictus mosquitoes. Half the horses were infected with a viral isolate from the brain of a horse (BC787), and half were infected with an isolate from crow brain (NY99-6625); both were NY99 isolates. Postinfection, uninfected female Ae. albopictus fed on eight of the infected horses. In the first trial, Nt antibody titers reached >1:320, 1:20, 1:160, and 1:80 for horses 1 to 4, respectively. In the second trial, the seven horses with subclinical infections developed Nt antibody titers >1:10 between days 7 and 11 post infection. The highest viremia level in horses fed upon by the recipient mosquitoes was approximately 460 Vero cell PFU/mL. All mosquitoes that fed upon viremic horses were negative for the virus. Horses infected with the NY99 strain of WNV develop low viremia levels of short duration; therefore, infected horses are unlikely to serve as important amplifying hosts for WNV in nature.

West Nile virus activity in Latin America and the Caribbean.

OBJECTIVES West Nile virus (Flavivirus: Flaviviridae; WNV) has spread rapidly throughout the Caribbean Basin since its initial detection there in 2001. This report summarizes our current knowledge of WNV transmission in tropical America. METHODS We reviewed the published literature and consulted with key public health officials to obtain unpublished data. RESULTS West Nile virus infections first appeared in human residents of the Cayman Islands and the Florida Keys in 2001, and in apparently healthy Jamaican birds sampled early in 2002. Serologic evidence of WNV infection in 2002 was detected in horses, chickens and resident free-ranging birds in Guadeloupe, the Dominican Republic, and eastern Mexico. In 2003, WNV spread in Mexico and northern Central America, and serologic evidence was detected in the Bahamas, Puerto Rico and Cuba. In 2004, the first serologic evidence of WNV activity in South American ecosystems surfaced in September-October in Colombia and Trinidad, where domestic animals circulated WNV-neutralizing antibodies. CONCLUSIONS The sparse reports of equine, human and avian disease in Latin America and the Caribbean is puzzling. Isolates are needed to evaluate viral attenuation or other possible explanations for reduced disease burden in tropical ecosystems.

Differential Virulence of West Nile Strains for American Crows

Increased viremia and deaths in American Crows inoculated with a North American West Nile viral genotype indicate that viral genetic determinants enhance avian pathogenicity and increase transmission potential of WNV.

Epitope-Blocking Enzyme-Linked Immunosorbent Assays for the Detection of Serum Antibodies to West Nile Virus in Multiple Avian Species

ABSTRACT We report the development of epitope-blocking enzyme-linked immunosorbent assays (ELISAs) for the rapid detection of serum antibodies to West Nile virus (WNV) in taxonomically diverse North American avian species. A panel of flavivirus-specific monoclonal antibodies (MAbs) was tested in blocking assays with serum samples from WNV-infected chickens and crows. Selected MAbs were further tested against serum samples from birds that represented 16 species and 10 families. Serum samples were collected from birds infected with WNV or Saint Louis encephalitis virus (SLEV) and from noninfected control birds. Serum samples from SLEV-infected birds were included in these experiments because WNV and SLEV are closely related antigenically, are maintained in similar transmission cycles, and have overlapping geographic distributions. The ELISA that utilized MAb 3.1112G potentially discriminated between WNV and SLEV infections, as all serum samples from WNV-infected birds and none from SLEV-infected birds were positive in this assay. Assays with MAbs 2B2 and 6B6C-1 readily detected serum antibodies in all birds infected with WNV and SLEV, respectively, and in most birds infected with the other virus. Two other MAbs partially discriminated between infections with these two viruses. Serum samples from most WNV-infected birds but no SLEV-infected birds were positive with MAb 3.67G, while almost all serum samples from SLEV-infected birds but few from WNV-infected birds were positive with MAb 6B5A-5. The blocking assays reported here provide a rapid, reliable, and inexpensive diagnostic and surveillance technique to monitor WNV activity in multiple avian species.

West Nile Virus in Livestock and Wildlife

WN virus is one of the most ubiquitous arboviruses occurring over a broad geographical range and in a wide diversity of vertebrate host and vector species. The virus appears to be maintained in endemic foci on the African continent and is transported annually to temperate climates to the north in Europe and to the south in South Africa. Reports of clinical disease due to natural WN virus infection in wild or domestic animals were much less common than reports of infection (virus isolation or antibody detection). Until recently, records of morbidity and mortality in wild birds were confined to a small number of cases and infections causing encephalitis, sometimes fatal, in horses were reported infrequently. In the period 1996-2001, there was an increase in outbreaks of illness due to WN virus in animals as well as humans. Within the traditional range of WN virus, encephalitis was reported in horses in Italy in 1998 and in France in 2000. The first report of disease and deaths caused by WN virus infection in domestic birds was reported in Israel in 1997-1999, involving hundreds of young geese. In 1999 WN virus reached North America and caused an outbreak of encephalitis in humans in the New York area at the same time as a number of cases of equine encephalitis and deaths in American crows and a variety of other bird species, both North American natives and exotics. Multi-state surveillance for WN virus has been in place since April 2000 and has resulted in the detection of WN virus in thousands of dead birds from an increasing number of species in North America, and also in several species of mammals. The surveillance system that has developed in North America because of the utility of testing dead birds for the rapid detection of WN virus presence has been a unique integration of public health and wildlife health agencies. It has been suggested that the recent upsurge in clinical WN virus infection in wild and domestic animals as well as in humans may be related to the emergence of one or more new strains of WN virus. Virus isolated in New York in 1999 was found to be identical to that from Israel. It was alarming for WN virus to so easily invade the United States and surprising that it became established so quickly in the temperature climate of New York. Its persistence and rapid expansion in the United States leave a number of unanswered questions. New disease characteristics and patterns have occurred and more are evolving as WN virus further invades the western hemisphere. Additional animal research is needed to answer these questions. Some of the research needs include bird migration as a mechanism of virus dispersal, vector and vertebrate host relationships, virus persistence mechanisms, laboratory diagnosis, viral pathogenesis, risk factor studies, vaccine development, and WN virus impact on wildlife (CDC 2001a). Determination of the primary reservoir host species that are involved in the epidemiology of WN virus and the suitable sentinel species for active surveillance are also important research areas.

Serologic evidence of West Nile virus infection in horses, Coahuila State, Mexico.

Serum samples were obtained from 24 horses in the State of Coahuila, Mexico, in December 2002. Antibodies to West Nile virus were detected by epitope-blocking enzyme-linked immunosorbent assay and confirmed by plaque reduction neutralization test in 15 (62.5%) horses. We report the first West Nile virus activity in northern Mexico.

Alligators as West Nile Virus Amplifiers

Juvenile alligators may help transmit West Nile virus in some areas.

Seasonal blood-feeding behavior of Culex tarsalis (Diptera: Culicidae) in Weld County, Colorado, 2007.

ABSTRACT Studies on Culex tarsalis Coquillett in Colorado have shown marked seasonal variation in the proportion of blood meals from birds and mammals. However, limitations in the specificity of antibodies used in the precipitin test and lack of vertebrate host availability data warrant revisiting Cx. tarsalis blood feeding behavior in the context of West Nile virus (WNV) transmission. We characterized the host preference of Cx. tarsalis during peak WNV transmission season in eastern Colorado and estimated the relative contribution of different avian species to WNV transmission. Cx. tarsalis preferred birds to mammals each month, although the proportion of blood meals from mammals increased in July and August. The distribution of blood meals differed significantly across months, in part because of changes in the proportion of blood meals from American robins, a preferred host. The estimated proportion of WNV-infectious vectors derived from American robins declined from 60 to 1% between June and August. The majority of avian blood meals came from doves, preferred hosts that contributed 25–40% of the WNV-infectious mosquitoes each month. Active WNV transmission was observed in association with a large house sparrow communal roost. These data show how seasonal patterns in Cx. tarsalis blood feeding behavior relate to WNV transmission in eastern Colorado, with the American robin contributing greatly to early-season virus transmission and a communal roost of sparrows serving as a focus for late-season amplification.

NATURAL AND EXPERIMENTAL WEST NILE VIRUS INFECTION IN FIVE RAPTOR SPECIES

We studied the effects of natural and/or experimental infections of West Nile virus (WNV) in five raptor species from July 2002 to March 2004, including American kestrels (Falco sparverius), golden eagles (Aquila chrysaetos), red-tailed hawks (Buteo jamaicensis), barn owls (Tyto alba), and great horned owls (Bubo virginianus). Birds were infected per mosquito bite, per os, or percutaneously by needle. Many experimentally infected birds developed mosquito-infectious levels of viremia (>105 WNV plaque forming units per ml serum) within5 days postinoculation (DPI), and/or shed virus per os or per cloaca. Infection of organs 15–27 days postinoculation was infrequently detected by virus isolation from spleen, kidney, skin, heart, brain, and eye in convalescent birds. Histopathologic findings varied among species and by method of infection. The most common histopathologic lesions were subacute myocarditis and encephalitis. Several birds had a more acute, severe disease condition represented by arteritis and associated with tissue degeneration and necrosis. This study demonstrates that raptor species vary in their response to WNV infection and that several modes of exposure (e.g., oral) may result in infection. Wildlife managers should recognize that, although many WNV infections are sublethal to raptors, subacute lesions could potentially reduce viability of populations. We recommend that raptor handlers consider raptors as a potential source of WNV contamination due to oral and cloacal shedding.

West Nile Virus Surveillance using Sentinel Birds

Abstract: Captive and free‐ranging birds have been used for decades as living sentinels in arbovirus surveillance programs. This review summarizes information relevant to selecting sentinel bird species for use in surveillance of West Nile (WN) virus. Although experience using avian sentinels for WN virus surveillance is limited, sentinels should be useful for both detecting and monitoring WN virus transmission; however, sentinel bird surveillance systems have yet to be adequately tested for use with the North American strain of WN virus. Captive chickens are typically used for arbovirus surveillance, but other captive species may be used as well. Serosurvey and experimental infection data suggest that both chickens and pigeons show promise as useful captive sentinels; both species were naturally exposed during the epizootics in New York City, 1999‐2000, and both species develop antibodies after infection without becoming highly infectious to Culex pipiens vectors. Wild bird species that should be targeted for use as free‐ranging sentinels include house sparrows and pigeons. The ideal wild bird should be determined locally on the basis of seroprevalence studies. Interpreting serological data generated from studies using free‐ranging sentinel birds is complex, however. Sentinel bird monitoring sites should be selected in enzootic transmission foci. Several years of observation may be required for selection of effective sentinel monitoring sites.