Infectivity of Wild-Bird Origin Influenza A Viruses in Minnesota Wetlands across Seasons.

The environmental tenacity of influenza A viruses (IAVs) in the environment likely plays a role in their transmission; IAVs are able to remain infectious in aquatic habitats and may have the capacity to seed outbreaks when susceptible wild bird hosts utilize these same environments months or even seasons later. Here, we aimed to assess the persistence of low-pathogenicity IAVs from naturally infected ducks in Northwestern Minnesota through a field experiment. Viral infectivity was measured using replicate samples maintained in distilled water in a laboratory setting as well as in filtered water from four natural water bodies maintained in steel perforated drums (hereafter, mesocosms) within the field from autumn 2020 to spring 2021. There was limited evidence for the extended persistence of IAVs held in mesocosms; from 65 initial IAV-positive samples, only six IAVs persisted to at least 202 days in the mesocosms compared to 17 viruses persisting at least this long when held under temperature-controlled laboratory settings in distilled water. When accounting for the initial titer of samples, viruses detected at a higher concentration at the initiation of the experiment persisted longer than those with a lower starting titer. A parallel experimental laboratory model was used to further explore the effects of water type on viral persistence, and the results supported the finding of reduced tenacity of IAVs held in mesocosms compared to distilled water. The results of this investigation provide evidence that many factors, including temperature and physicochemical properties, impact the duration of viral infectivity in natural settings, further extending our understanding of the potential and limitations of environmental-based methodologies to recover infectious IAVs.

Distribution and prevalence of antibodies to Trichinella spp. and Toxoplasma gondii in wild pigs (Sus scrofa) in the United States.

Invasive wild pigs (Sus scrofa) are a reservoir for over 100 viral, bacterial, and parasitic pathogens that are transmissible to humans, livestock, domestic animals, and wildlife in North America. Numerous historical local surveys and results from a nation-wide survey (2006-2010) indicated that wild pigs in the United States act as reservoirs for Trichinella spp. and Toxoplasma gondii, two zoonotic pathogens of importance for human and animal health. Since that time, wild pig populations have expanded and increased in density in many areas. Population expansion of wild pigs creates opportunities for the introduction of pathogens to new areas of the country, increasing health risks. The goal of this study was to investigate the current geographic distribution and prevalence of Trichinella spp. and T. gondii antibodies in wild pigs using serum samples collected from 2014 to 2020. Serum samples from 36 states were tested for antibodies to Trichinella spp. (n = 7467) and T. gondii (n = 5984) using commercially available enzyme-linked immunosorbent assays. Seroprevalence for Trichinella spp. (12.4%, 927/7467) and T. gondii (40.8%, 2444/5984) are significantly higher compared to a previous 2006-2010 study across all regions. Results from this study also showed a lower seroprevalence (4.8%) for Trichinella spp. in the West region compared to the other regions (South: 13.4%; Midwest: 18.4%; Northeast: 19.1%). There were new detection records for antibodies to Trichinella spp. in 11 states, mostly in the West, Midwest, and Northeast regions compared to a previous study in 2014. Males and juveniles were less likely to be positive for Trichinella spp. antibodies, compared to females and older animals, respectively. Seroprevalence was similar for T. gondii across the regions (31.8-56%) with some states having particularly high seroprevalence (e.g., Hawaii 79.4% and Pennsylvania 68%). There were new T. gondii antibody detection records for 12 states, mostly in the West, Midwest, and Northeast regions. Adults were more likely than juveniles and subadults to be seropositive. These data confirm that the distribution and prevalence of antibodies for Trichinella spp. and T. gondii are increasing in the United States, likely driven by wild pig population growth and range expansion.

Bioaccumulation of Some Metals and Metalloids in Laughing Gulls (Leucophaeus atricilla): Increases in Mercury and Decreases in Selenium from 2019 to 2022/2023.

The elements in blood normally reflect the levels in prey, indicating a recent exposure. Laughing gulls (Leucophaes atricilla) eat mainly horseshoe crab eggs (Limulus polyphemus) in the spring in Delaware Bay, New Jersey. The levels of arsenic (As), cadmium (Cd), chromium (Cr), lead (Pb), mercury (Hg), and selenium (Se) in the blood of laughing gulls foraging on crab eggs were examined in Delaware Bay to provide information on a species that is normally a generalist, and to determine if the levels of these elements were similar in 2019 and 2022/2023, were intercorrelated, and were related to those in crab eggs. Hg increased from 2019 (136 ± 31 ng/g) to 2022/2023 (473 ± 75 ng/g), while Cd and Se decreased. There were some significant correlations among elements and a close relationship between the element levels in blood and those in crab eggs collected in the same month (except for As). The levels differed between laughing gulls and three species of shorebirds for As and Cd. The elements in the blood of gulls and shorebirds should be similar because they eat mainly the same eggs in the same places. A significant proportion of laughing gull blood samples had levels of Hg and Se that were above the levels associated with adverse effects, which requires further examination.

Naturally Acquired Antibodies to Influenza A Virus in Fall-Migrating North American Mallards.

Although waterfowl are the primary reservoir for multiple subtypes of influenza A virus (IAV), our understanding of population immunity in naturally infected waterfowl is poorly understood. Population immunity may be an important driver of seasonal subtype predominance in waterfowl populations and may affect the potential for establishment of introduced IAV such as the Eurasian-like A/Goose/Guangdong/1/1996 lineage in these populations. Here, we examine the prevalence of naturally acquired antibodies to nucleoprotein (NP), hemagglutinin (H3, H4, H5), and neuraminidase (N1, N2, N6, N8) in early migrating mallards (Anas platyrhynchos) sampled in Northwest Minnesota during staging and early fall migration in September 2014, 2015, 2017, and 2018. Serologic results were compared to historic and contemporary virus isolation results from these same study sites. The prevalence of antibodies to NP ranged from 60.8−76.1% in hatch-year (HY) birds and from 86.0−92.7% in after-hatch-year (AHY, >1-year-old) mallards indicating a high level of previous infection with IAV early in the fall migration season. Neutralizing antibodies were detected against H3, H4, and H5 in all years as were antibodies to N1, N2, N6, and N8. A high proportion of NP seropositive ducks tested positive for antibodies to multiple HA and NA subtypes, and this was more common in the AHY age class. Antibody prevalence to the HA and NA subtypes included in this study were consistent with the predominance of H4N6 in these populations during all years and reflected a broadening of the antibody response with age. Additional work is needed to document the longevity of these immune responses, if and how they correlate with protection against IAV transmission, infection, and disease, and if, as detected in this study, they adequately describe the true extent of exposure to IAV or specific HA or NA subtypes.

The Effect of Maternal Antibodies on Clinical Response to Infection with Epizootic Hemorrhagic Disease Virus in White-Tailed Deer (Odocoileus virginianus) Fawns.

We investigated whether naturally acquired maternal antibodies to epizootic hemorrhagic disease virus serotype 2 (EHDV-2) would protect white-tailed deer (Odocoileus virginianus) fawns against infection and clinical disease following an EHDV-2 challenge. We compared viremia and clinical response in 27-47-d-old, experimentally infected fawns with and without maternally derived antibodies to EHDV-2. Mild to moderate clinical signs were observed in four seronegative (maternal antibody-negative) fawns, which were viremic from 3 to 14 d postinoculation. Individual peak blood virus titers for seronegative fawns ranged from 104.3 to 106.3 median tissue culture infective doses (TCID50)/mL. In contrast, clinical signs were not observed in seropositive (maternal antibody-positive) fawns and a transient low-level viremia (≤102.4 TCID50/mL) occurred in two of six fawns. Our results indicated that the presence of maternally derived EHDV-2 antibodies in fawns prevents or greatly reduces clinical disease and the level and duration of EHDV-2 viremia.

Prevalence of Influenza A Viruses in Ducks Sampled in Northwestern Minnesota and Evidence for Predominance of H3N8 and H4N6 Subtypes in Mallards, 2007-2016.

Long-term comprehensive studies of avian influenza virus subtypes in ducks not only contribute to understanding variations and patterns of subtype diversity, but also can be important in defining seasonal and temporal risks associated with transmission of potentially highly pathogenic H5 and H7 subtypes to domestic poultry. We analyzed influenza A virus (IAV) surveillance data from dabbling ducks collected at an important migratory stopover site in northwestern Minnesota from 2007-2016 and identified prevalence and subtype diversity throughout this period. In total, 13,228 cloacal and oropharyngeal swabs from waterfowl were tested over the 10-year period; the majority of these waterfowl were mallards sampled from late August through late September (n = 9133). From these, 1768 IAVs were isolated (19.4% mean annual prevalence, ranging from 11.0% in 2007 to 32.8% in 2011), and both hemagglutinin (HA) and neuraminidase were identified for 1588. Although subtype diversity and prevalence varied by year, H3 and H4 HA subtypes predominated in all years, accounting for 65.7% of the observed HA subtype diversity. The mechanisms driving this consistent pattern of subtype diversity and predominance are not understood but may include factors at the host, population, and virus level.

Prevalencia de virus de influenza A en patos muestreados en el noroeste de Minnesota y evidencia de predominio de los subtipos H3N8 y H4N6 en patos de collar entre los años 2007 al 2016. Los estudios exhaustivos a largo plazo de subtipos de virus de la influenza aviar en patos no solo contribuyen a comprender las variaciones y patrones de diversidad de subtipos, sino que también pueden ser importantes para definir los riesgos estacionales y temporales asociados con la transmisión de subtipos H5 y H7 potencialmente altamente patógenos para la avicultura comercial. Analizamos los datos de vigilancia del virus de la influenza A de patos chapoteadores recolectados en un sitio de descanso migratorio importante en el noroeste de Minnesota desde el año 2007 al 2016 y se identificó la prevalencia y la diversidad de subtipos a lo largo de este período. En total, 13,228 hisopos cloacales y orofaríngeos de aves acuáticas se analizaron durante el período de diez años; la mayoría de estas aves acuáticas eran patos silvestres muestreados desde finales de agosto hasta finales de septiembre (n = 9133). De estas muestras, 1768 virus de la influenza aviar fueron aislados (prevalencia anual media de 19.4%, y con un rango de 11.0% en el 2007 a 32.8% en 2011), y tanto la hemaglutinina (HA) como la neuraminidasa fueron identificadas para 1588 virus. Aunque la diversidad de subtipos y la prevalencia variaron por año, los subtipos de hemaglutinina H3 y H4 predominaron en todos los años, representando el 65.7% de la diversidad de subtipos observada para la hemaglutinina. Los mecanismos que impulsan este patrón consistente de diversidad de subtipos y predominio no se comprenden, pero pueden incluir factores a nivel del hospedador, de la población y del virus. Abbreviations: CL = cloacal; HA = hemagglutinin; IAV = influenza A virus; NA = neuraminidase; NWR = National Wildlife Refuge; RRT-PCR = real-time reverse transcriptase PCR; WMA = Wildlife Management Area.

Competition between influenza A virus subtypes through heterosubtypic immunity modulates re-infection and antibody dynamics in the mallard duck.

Our overall hypothesis is that host population immunity directed at multiple antigens will influence the prevalence, diversity and evolution of influenza A virus (IAV) in avian populations where the vast subtype diversity is maintained. To investigate how initial infection influences the outcome of later infections with homologous or heterologous IAV subtypes and how viruses interact through host immune responses, we carried out experimental infections in mallard ducks (Anas platyrhynchos). Mallards were pre-challenged with an H3N8 low-pathogenic IAV and were divided into six groups. At five weeks post H3N8 inoculation, each group was challenged with a different IAV subtype (H4N5, H10N7, H6N2, H12N5) or the same H3N8. Two additional pre-challenged groups were inoculated with the homologous H3N8 virus at weeks 11 and 15 after pre-challenge to evaluate the duration of protection. The results showed that mallards were still resistant to re-infection after 15 weeks. There was a significant reduction in shedding for all pre-challenged groups compared to controls and the outcome of the heterologous challenges varied according to hemagglutinin (HA) phylogenetic relatedness between the viruses used. There was a boost in the H3 antibody titer after re-infection with H4N5, which is consistent with original antigenic sin or antigenic seniority and suggest a putative strategy of virus evasion. These results imply competition between related subtypes that could regulate IAV subtype population dynamics in nature. Collectively, we provide new insights into within-host IAV complex interactions as drivers of IAV antigenic diversity that could allow the circulation of multiple subtypes in wild ducks.

Antibodies to Influenza A Viruses in Wintering Snow Geese (Chen caerulescens) in Texas.

Wild waterfowl in the order Anseriformes are recognized reservoirs for influenza A viruses (IAVs); however, prevalence of infection can vary greatly by species. Few isolates of IAVs have been reported from snow geese (Chen caerulescens), and generally they have not been regarded as an important component of this reservoir. In February 2013, 151 combined cloacal and oropharangeal swabs and 147 serum samples were collected from snow geese wintering on the Gulf coast of Texas. None of the swab samples tested positive by virus isolation, but antibodies to IAVs were detected in 87 (59%) birds tested by competitive blocking ELISA (bELISA). To further characterize these detected antibodies, positive samples were tested by virus microneutralization (MN) for antibodies to viruses representing 14 hemagglutinin subtypes (HA1-HA12, H14, and H15). By MN, antibodies to H1 (n = 41; 47%), H5 (n = 32; 37%), H6 (n = 49; 56%), H9 (n = 50; 57%), and H12 (n = 24; 28%) were detected. Snow goose populations have increased in North America since the 1960s, and their association with agricultural lands provides a potential indirect source of IAV infection for domestic poultry. This potential, as well as the detection of antibodies to HA subtypes H5, H9, and H12 that are not well represented in other waterfowl species, suggests that further snow geese surveillance is indicated.

Antibodies to Influenza A Viruses in Gulls at Delaware Bay, USA.

Gulls are the known reservoir for H13 and H16 influenza A viruses (IAV) but also host a diversity of other IAV subtypes. Gulls also share habitats with both ducks and shorebirds, increasing the potential for cross-species IAV transmission. We serologically tested laughing gulls (Leucophaeus atricilla) collected at Delaware Bay during May when they were in direct contact with IAV-infected shorebirds; both species feed on horseshoe crab (Limulus polyphemus) eggs on beaches during this month. From 2010 to 2014, antibody prevalence as determined by competitive blocking enzyme-linked immunosorbent assay ranged from 25%-72%. Antibodies to H13 and H16 were detected by hemagglutination inhibition (HI) tests in 12% and 24% of tested gulls, respectively. Results from virus microneutralization (MN) tests for antibodies to H1-H12, H14, and H15 varied among years but the highest prevalence of neutralizing antibodies was detected against H1 (24%), H5 (25%), H6 (35%), H9 (33%), and H11 (42%) IAV. The subtype diversity identified by serology in gulls was dominated by Group 1 HA subtypes and only partially reflected the diversity of IAV subtypes isolated from shorebirds.