West Nile Virus Mortality Research Articles

Christmas Bird Count data are a reliable harvest-independent index for Ruffed Grouse monitoring in the eastern United States

Abstract Using data from the mid-1990s to the present, we assessed correlations of Ruffed Grouse (Bonasa umbellus) hunter cooperator reports from seven states to Breeding Bird Survey, Christmas Bird Count, and eBird data to determine which program’s trends best reflected patterns seen in hunter-sourced data that has historically been used for monitoring purposes. Christmas Bird Count data most closely reflected state-wide long-term trends in cooperator flush rates. Using an analytical approach that accounts for sources of bias and examines abundance at a finer spatial scale, we observed a pattern of decline across our study area. In recent decades, Ruffed Grouse populations in the eastern United States are thought to have suffered persistent declines due to a combination of young forest habitat loss and West Nile virus mortality. Declining hunter participation in much of the species’ range has highlighted the need to identify harvest-independent indices of grouse abundance to enable continued monitoring of regional long-term trends. Recent analytical methods can better capture complex trends in population dynamics, account for biases inherent to community-sourced data, and potentially inform designs of future survey efforts aimed at fulfilling priority monitoring and research needs. Comparable results across varying statistical methodologies suggest that our inferences are robust, demonstrating that Christmas Bird Count data can be used to inform regional monitoring efforts for grouse and other cryptic game bird populations, but require careful consideration of program designs and protocols to identify and implement data appropriately.

WEST NILE VIRUS EXPOSURE AND INFECTION AMONG HUNTER-HARVESTED RUFFED GROUSE (BONASA UMBELLUS) COHORTS IN A STABLE POPULATION.

West Nile virus (WNV) was introduced to North America two decades ago, but for many species, including Ruffed Grouse (Bonasa umbellus), the effects of WNV on individuals and populations remain poorly understood. Recent studies suggest the effect of WNV on Ruffed Grouse might vary among geographic regions, depending on habitat conditions. We studied WNV in Minnesota, US, during 2018-19, in a region known to have abundant Ruffed Grouse habitat and a population cycling around a stable long-term average. We worked with cooperating hunters to collect hearts, feathers, and blood on filter strips from birds harvested in the fall to examine exposure to the virus. We detected antibodies to WNV or a flavivirus (probably WNV) in 12.5% and 12.3% of birds in 2018 and 2019, respectively. However, we did not isolate the virus from any heart samples, indicating that exposed birds were not experiencing an active infection of WNV at the time of harvest. Our findings indicate that, although Minnesota Ruffed Grouse are exposed to WNV, some birds mount a successful immune response and survive. However, our sampling approach did not account for birds that might have become infected over the summer and died, so it is unknown how much WNV mortality occurred before the fall harvest. Birds lost to WNV over the summer could reduce the number of birds that hunters see in the fall, thus reducing the quality of their hunting experiences. Management options for mitigating WNV impacts and other stressors consist primarily of providing high-quality Ruffed Grouse habitat that produces birds in good condition that are more likely to recover from infection.

Experimental susceptibility of wood ducks (Aix sponsa) for West Nile virus.

Detection of West Nile virus (WNV) has been reported in a variety of wild ducks in the US, but little is known about the pathogenesis and outcome of exposure of the disease in these species. Previous experimental studies of WNV in ducks either have challenged a small number of ducks with WNV or have tested domesticated ducks. To determine susceptibility and immune response, we challenged 7-wk-old Wood Ducks (Aix sponsa) with a 1999 American Crow (Corvus brachyrhynchos) isolate of WNV. Wood Ducks were susceptible to infection with the virus, and, although clinical signs or mortality were not observed, microscopic lesions were noted, particularly in the heart and brain. West Nile virus viremia peaked on day 2 postinfection (pi) at 10(4.54) plaque-forming units (PFU) of virus/mL serum and WNV was shed orally (between 10(2) and 10(2.9) PFU per swab) and cloacally. Specific anti-WNV antibody response was rapid, with anti-WNV IgM detected on day 3 pi followed on day 5 pi by anti-WNV IgG. Neutralizing antibodies were detected by plaque-reduction neutralization assay in one duck on day 4 pi, and in all sampled ducks on day 5. These results indicate that Wood Ducks are susceptible to WNV, but it is unlikely that significant WNV mortality events occur in Wood Ducks or that they play a significant role in transmission. However, WNV viremia was sufficient, in theory, to infect mosquitoes, and oral and cloacal shedding of the virus may increase the risk of infection to other waterbirds.

Predicted and observed mortality from vector-borne disease in wildlife: West Nile virus and small songbirds

Numerous diseases of wildlife have recently emerged due to trade and travel. However, the impact of disease on wild animal populations has been notoriously difficult to detect and demonstrate, due to problems of attribution and the rapid disappearance of bodies after death. Determining the magnitude of avian mortality from West Nile virus (WNV) is emblematic of these challenges. Although correlational analyses have shown population declines coincident with the arrival of the virus, strong inference of WNV as a cause of mortality or a population decline requires additional evidence. We show how integrating field data on mosquito feeding patterns, avian abundance, and seroprevalence can be used to predict relative mortality from vector-borne pathogens. We illustrate the method with a case study on WNV in three species of small songbirds, tufted titmouse (Baeolophus bicolor), Carolina wrens (Thryothorus ludovicianus), and northern cardinals (Cardinalis cardinalis). We then determined mortality, infectiousness, and behavioral response of wrens and titmouse following infection with WNV in laboratory experiments and compared them to a previous study on WNV mortality in cardinals. In agreement with predictions, we found titmouse had the highest mortality from WNV infection, with 100% of 11 birds perishing within 7days after infection. Mortality in wrens was significantly lower at 27% (3/11), but still substantial. Viremia profiles indicated that both species were highly infectious for WNV and could play roles in WNV amplification. These findings suggest that WNV may be killing many small-bodied birds, despite the absence of large numbers of dead birds being observed and testing positive for WNV. More broadly, they illustrate the utility of a framework for predicting relative mortality in hosts from vector-borne disease.

West Nile Virus in American White Pelican Chicks: Transmission, Immunity, and Survival

West Nile virus (WNV) causes significant mortality of American White Pelican chicks at northern plains colonies. We tested oropharyngeal/cloacal swabs from moribund chicks for shed WNV. Such shedding could enable chick-to-chick transmission and help explain why WNV spreads rapidly in colonies. WNV was detected on swabs from 11% of chicks in 2006 and 52% of chicks in 2007; however, viral titers were low. Before onset of WNV mortality, we tested blood from < 3-week-old chicks for antibodies to WNV; 5% of chicks were seropositive, suggesting passive transfer of maternal antibodies. Among near-fledged chicks, 41% tested positive for anti-WNV antibodies, indicating that they survived infection. Among years and colonies, cumulative incidence of WNV in chicks varied from 28% to 81%, whereas the proportion of chicks surviving WNV (i.e., seropositive) was 64-75%. Our data revealed that WNV kills chicks that likely would fledge in the absence of WNV, that infection of chicks is pervasive, and that significant numbers of chicks survive infection.

Field-Based Estimates of Avian Mortality from West Nile Virus Infection

One of the unique characteristics of West Nile virus (WNV) in North America is the large number of bird species for which the virus can be fatal. WNV mortality has been documented through experimental infections of captive birds and necropsies of free-ranging birds. Investigations of WNV-related mortality in wild birds often focus on species with dramatic population declines (e.g., American Crow, Corvus brachyrhynchos); however, few studies have addressed WNV-related mortality in species not exhibiting marked population declines since the arrival of WNV. We conducted a mark-recapture study of 204 Northern Cardinals (Cardinalis cardinalis) in an area with endemic WNV activity to estimate WNV-related mortality. Previous research has shown that once a bird is infected and recovers from WNV it develops antibodies making it resistant to future infection. Assuming that mortality risks from non-WNV causes were the same for individuals with (had been exposed to WNV) and without antibodies (had not been exposed to WNV), we compared the survival rates of birds with and without WNV antibodies to estimate the impact of WNV on wild birds. An information theoretic approach was used, and the apparent survival was found to be 34.6% lower for individuals without antibodies during the period when WNV was most active (July-September). However, the apparent survival rate was 9.0% higher for individuals without antibodies over the rest of the year. These differences in apparent survival suggest that WNV increases mortality during the WNV season and that chronic effects of WNV infection may also be contributing to mortality. Although WNV appears to have increased mortality rates within the population, population trend data do not indicate declines, suggesting that some cardinal populations can compensate for WNV-related mortality.

RISK FACTORS ASSOCIATED WITH WEST NILE VIRUS MORTALITY IN AMERICAN CROW POPULATIONS IN SOUTHERN QUEBEC

Soon after the appearance of West Nile virus (WNV) in North America, a number of public health authorities designated the American Crow (Corvus brachyrhynchos) a sentinel for WNV detection. Although preliminary studies have suggested a positive association between American Crow mortality and increased risk of WNV infection in humans, we still know little about dynamic variation in American Crow mortality, both baseline levels and mortality associated with WNV. We hypothesized that the complex social behavior of American Crows, which is shaped by age and seasonal factors, influences both baseline mortality and WNV mortality in American Crow populations. We examined American Crow mortality data from Quebec for the 2005 WNV surveillance year, which lasted from 5 June to 17 September 2005. The variables of interest were age, gender, body condition index, time of year, and land cover. We used a log-linear model to examine baseline mortality. Logistic regression and general linear regression models were constructed to examine variables associated with mortality due to WNV. We found that both age and time of year were key variables in explaining baseline mortality. These two variables were also risk factors for WNV mortality. The probability that a carcass tested positive for WNV increased with the age of the dead bird and as summer progressed. WNV-positive carcasses also had a lower body condition index than WNV-negative carcasses. We believe that the first major wave of American Crow mortality observed in the early summer of 2005 was the result of natural mortality among young American Crows. Because this mortality was not linked to WNV, it appears that American Crow may not be a good species for early detection of WNV activity. Our data also suggest that second-year American Crows play a major role in propagating WNV during their movements to urban land covers during midsummer.

Toll-Like Receptor 3 Has a Protective Role against West Nile Virus Infection

Protection against West Nile virus (WNV) infection requires rapid viral sensing and the generation of an interferon (IFN) response. Mice lacking IFN regulatory factor 3 (IRF-3) show increased vulnerability to WNV infection with enhanced viral replication and blunted IFN-stimulated gene (ISG) responses. IRF-3 functions downstream of several viral sensors, including Toll-like receptor 3 (TLR3), RIG-I, and MDA5. Cell culture studies suggest that host recognizes WNV in part, through the cytoplasmic helicase RIG-I and to a lesser extent, MDA5, both of which activate ISG expression through IRF-3. However, the role of TLR3 in vivo in recognizing viral RNA and activating antiviral defense pathways has remained controversial. We show here that an absence of TLR3 enhances WNV mortality in mice and increases viral burden in the brain. Compared to congenic wild-type controls, TLR3(-/-) mice showed relatively modest changes in peripheral viral loads. Consistent with this, little difference in multistep viral growth kinetics or IFN-alpha/beta induction was observed between wild-type and TLR3(-/-) fibroblasts, macrophages, and dendritic cells. In contrast, a deficiency of TLR3 was associated with enhanced viral replication in primary cortical neuron cultures and greater WNV infection in central nervous system neurons after intracranial inoculation. Taken together, our data suggest that TLR3 serves a protective role against WNV in part, by restricting replication in neurons.

Investigating the spatial risk distribution of West Nile virus disease in birds and humans in southern Ontario from 2002 to 2005.

BackgroundThe West Nile virus (WNv) became a veterinary public health concern in southern Ontario in 2001 and has continued to threaten public health. Wild bird mortality has been shown to be an indicator for tracking the geographic distribution of the WNv. The purpose of this study was to investigate the latent risk distribution of WNv disease among dead birds and humans in southern Ontario and to compare the spatial risk patterns for the period 2002–2005. The relationship between the mortality fraction in birds and incidence rate in humans was also investigated.MethodsChoropleth maps were created to investigate the spatial variation in bird and human WNv risk for the public health units of southern Ontario. The data were smoothed by empirical Bayesian estimation before being mapped. Isopleth risk maps for both the bird and human data were created to identify high risk areas and to investigate the potential relationship between the WNv mortality fraction in birds and incidence rates in humans. This was carried out by the geostatistical prediction method of kriging. A Poisson regression analysis was used to model regional human WNv case counts as a function of the spatial coordinates in the east and north direction and the regional bird mortality fractions. The presence of disease clustering and the location of disease clusters were investigated by the spatial scan test.ResultsThe isopleth risk maps exhibited high risk areas that were relatively constant from year to year. There was an overlap in the bird and human high risk areas, which occurred in the central-west and south-west areas of southern Ontario. The annual WNv cause-specific mortality fractions in birds for 2002 to 2005 were 31.9, 22.0, 19.2 and 25.2 positive birds per 100 birds tested, respectively. The annual human WNv incidence rates for 2002 to 2005 were 2.21, 0.76, 0.13 and 2.10 human cases per 100,000 population, respectively. The relative risk of human WNv disease was 0.72 times lower for a public health unit that was 100 km north of another public health unit. The relative risk of human WNv disease increased by the factor 1.44 with every 10 positive birds per 100 tested. The scan statistic detected disease cluster in the bird and human data. The human clusters were not significant, when the analysis was conditioned on the bird data.ConclusionThe study indicates a significant relationship between the spatial pattern of WNv risk in humans and birds.

PATHOLOGY AND EPIDEMIOLOGY OF NATURAL WEST NILE VIRAL INFECTION OF RAPTORS IN GEORGIA

Carcasses from 346 raptors found between August 2001 and December 2004 were tested for West Nile virus (WNV) using virus isolation and immunohistochemistry; 40 were positive for WNV by one or both methods. Of these 40 birds, 35 had histologic lesions compatible with WNV infection, one had lesions possibly attributable to WNV, and four had no histologic evidence of WNV. The most common histologic lesions associated with WNV infection were myocardial inflammation, necrosis, and fibrosis; skeletal muscle degeneration, inflammation, and fibrosis; and lymphoplasmacytic encephalitis. Other lesions included hepatitis, lymphoid depletion in spleen and bursa, splenic and hepatic hemosiderosis, pancreatitis, and ganglioneuritis. Gross lesions included calvarial and leptomeningeal hemorrhage, myocardial pallor, and splenomegaly. Red-tailed hawks (Buteo jamaicensis) (10/56), sharp-shinned hawks (Accipiter striatus) (8/40), and Cooper's hawks (Accipiter cooperii) (10/103) were most commonly affected. Also affected were red-shouldered hawks (Buteo lineatus) (2/43), an osprey (Pandion haliaetus) (1/5), barred owls (Strix varia) (4/27), a great horned owl (Bubo virginianus) (1/18), and eastern screech owls (Megascops asio) (4/42). Although birds were examined throughout the year, positive cases occurred only during the summer and late fall (June-December). Yearly WNV mortality rates ranged from 7-15% over the four years of the study. This study indicates trends in infection rates of WNV in raptorial species over a significant time period and supports the available information regarding pathology of WNV infection in Strigiformes and Falconiformes. Although many species tested were positive for WNV infection, severity of lesions varied among species.

Castanospermine, a Potent Inhibitor of Dengue Virus Infection In Vitro and In Vivo

Previous studies have suggested that alpha-glucosidase inhibitors such as castanospermine and deoxynojirimycin inhibit dengue virus type 1 infection by disrupting the folding of the structural proteins prM and E, a step crucial to viral secretion. We extend these studies by evaluating the inhibitory activity of castanospermine against a panel of clinically important flaviviruses including all four serotypes of dengue virus, yellow fever virus, and West Nile virus. Using in vitro assays we demonstrated that infections by all serotypes of dengue virus were inhibited by castanospermine. In contrast, yellow fever virus and West Nile virus were partially and almost completely resistant to the effects of the drug, respectively. Castanospermine inhibited dengue virus infection at the level of secretion and infectivity of viral particles. Importantly, castanospermine prevented mortality in a mouse model of dengue virus infection, with doses of 10, 50, and 250 mg/kg of body weight per day being highly effective at promoting survival (P < or = 0.0001). Correspondingly, castanospermine had no adverse or protective effect on West Nile virus mortality in an analogous mouse model. Overall, our data suggest that castanospermine has a strong antiviral effect on dengue virus infection and warrants further development as a possible treatment in humans.

Outbreak of West Nile virus in greater sagegrouse and guidelines for monitoring, handling, and submitting dead birds

West Nile virus (WNV) resulted in a 25% decline in survival in four populations of radiomarked greater sage-grouse (Centrocercus urophasianus) across Alberta, Wyoming, and Montana in 2003. Unexpected impacts of WNV are disturbing because range-wide habitat loss and degradation already threaten sage-grouse populations. In the Powder River Basin of Wyoming and Montana, late-summer survival of sage-grouse was lower at a site with confirmed WNV mortalities (20%) than at two sites without (76%). Dramatic declines in both male and female lek attendance at the WNV site the following spring suggest that outbreaks may threaten some local populations with extirpation. The key to understanding broader impacts of WNV on sage-grouse is to monitor additional populations and to determine whether populations infected in 2003 are again impacted this year. To facilitate this process, we describe a strategy for monitoring WNV mortality in the field and provide information on how to handle, store, and submit dead birds for testing.