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David E. Stallknecht

Publications and source records attributed to David E. Stallknecht.

At least 19 recordsLinked to original sources

Rapid expansion of genotype D1.1 A(H5N1) influenza viruses in wild birds across North America during the 2024 migratory season

In late 2021, high pathogenicity avian influenza A(H5N1) clade 2.3.4.4b viruses entered North America and reassorted rapidly with local avian influenza viruses. In September 2024, we detected a new reassortant later classified as genotype D1.1. Using active and passive avian influenza surveillance across Canada and the USA, we tracked the emergence and rapid spread of D1.1 viruses in wild birds during the 2024 fall migration. Phylodynamic analysis showed that D1.1 viruses formed a monophyletic group and displaced earlier A(H5) genotypes across several flyways. Their expansion coincided with detections in other hosts, including 17 human cases, 4 of which were severe or fatal. None of the mammalian-adaptive markers detected in human cases were found in wild bird viruses, and candidate vaccine viruses retained antigenic cross-reactivity with D1.1 strains.

Nature medicine

Rapid increase in antibodies to influenza A virus H5 and N1 in Lesser Scaup (Aythya affinis) following the introduction of 2.3.4.4B H5N1 into North America

Clade 2.3.4.4b Eurasian-origin H5N1 entered North America in late 2021 and spread across the continent. While studies have characterized the antibody response mounted by dabbling ducks following exposure, little data are available for diving ducks. This study sought to identify influenza A virus (IAV) infection and antibodies in Lesser and Greater Scaup captured in Maryland, Illinois, and Rhode Island. In Maryland, IAV seroprevalence increased from the 2021/2022 to 2022/2023 sampling season, with IAV antibody prevalence increasing for juvenile (38% to 80%) and adult (82% to 90%) Lesser Scaup. While adult Lesser Scaup sampled in Illinois in 2021/2022 had IAV antibody prevalence comparable to those sampled in Maryland (76% and 82%, respectively), they had higher antibody prevalence to both H5 (48% and 18%) and N1 (68% and 35%), potentially due to being sampled in March versus December and January. Our data suggest that Lesser Scaup had limited antibodies to highly pathogenic H5 IAV prior to the introduction of clade 2.3.4.4b H5N1 to North America, but relevant antibodies were widely observed in the months and year following. Our more limited data suggest similar trends may have occurred in Greater Scaup as well.

Canadian Journal of Microbiology

Potential impacts of 2.3.4.4b highly pathogenic H5N1 avian influenza virus infection on Snow Goose (Anser caerulescens) movement ecology

While wild waterfowl are known reservoirs of avian influenza viruses and facilitate the movement of these viruses, there are notable differences in the response to infection across species. This study explored differential responses to infection with highly pathogenic avian influenza in Snow Geese ( Anser caerulescens ) located in the California Central Valley. Though H5 antibody prevalence was high across years among birds sampled in the winter (75% in both years via hemagglutination inhibition), these values were even higher among birds sampled in summer that failed to migrate (i.e., August 2023 = 100% and August 2024 = 93% via hemagglutination inhibition). Birds that failed to migrate were also generally lighter than birds sampled in the winter and presented notable damage to cerebrum and cerebellum. In December 2022, a single individual positive for infection with H5N1 at the time of sampling indicated reduced movement during the 14 days following sampling but completed spring migration comparably with uninfected conspecifics. However, while no birds were actively infected during sampling and marking in 2023, two marked geese departed for migration late and one did not migrate at all. Additional banded birds marked in August have been reencountered in scenarios ranging from hunter harvest at a different site over a year later to found dead shortly after banding. Our data indicate that Snow Geese infected with HPAI have the potential to express variable outcomes following infection with highly pathogenic H5N1, ranging from rapid recovery within a migratory season to death. These data also suggest that the abnormal failure of some Snow Geese to migrate from the Central Valley is likely driven by HPAI infection.

California

Avian influenza infection and antibodies not identified in eastern wild turkeys (Meleagris gallopavo silvestris) sampled on the Delmarva Peninsula

Although highly pathogenic avian influenza presents a notable threat to domestic turkeys ( Meleagris gallopavo domesticus ), there have been minimal infections identified in wild turkeys ( Meleagris gallopavo ) across the United States. Indeed, recent infections in wild turkeys have either been linked back to or suspected to be the result of spillback from nearby infected domestic turkeys. This study presents avian influenza sampling data for Eastern wild turkeys ( Meleagris gallopavo silvestris ) across the Maryland portion of the Delmarva Peninsula, in the winter of 2023–2024, with an objective of identifying any circulating influenza A viruses (IAVs), as well as determining the extent of previous exposure to IAVs in this population. We collected swab samples from 36 female wild turkeys. No IAV viral RNA was detected in any sample, based on real-time reverse transcriptase PCR. We collected sera from 31 of the 36 sampled individuals, all of which had biomimetic enzyme-linked immunosorbent assay (bELISA) S/N values >0.7 (range: 0.79–1.29) indicating a lack of antibodies to IAVs. Although our sample size is limited, these results indicate no evidence of current or recent previous IAV infection in sampled birds.

Maryland

Exposure of wild mammals inhabiting Alaska to influenza A(H5N1) virus

Serum samples from wild mammals inhabiting Alaska, USA, showed that 4 species, including Ursus arctos bears and Vulpes vulpes foxes, were exposed to influenza A(H5N1) viruses. Results indicated some mammals in Alaska survived H5N1 virus infection. Surveillance efforts may be improved by incorporating information on susceptibility and detectable immune responses among wild mammals.

Alaska

Highly pathogenic avian influenza virus H5N1 in double-crested cormorants (Nannopterum auritum) of the Chesapeake Bay, USA

Double-crested Cormorants ( Nannopterum auritum ) have historically exhibited low levels of infection and antibodies to avian influenza virus (AIV). The recent global expansion of clade 2.3.4.4b A/goose/Guangdong/1/1996 highly pathogenic (HP) avian influenza virus H5N1 (HPAI H5N1) has resulted in large-scale mortalities across diverse waterbird taxa including cormorants. We sampled 32 and 29 Double-crested Cormorants breeding in the Chesapeake Bay, US, during the summers of 2023 and 2024, respectively, to assess HPAI H5N1 infection and AIV antibodies. Although no mortality was observed in the area, one bird sampled in 2023 was infected with HPAI H5N1. Additionally, 21/31 individuals in 2023 and 10/25 individuals in 2024 for which sera were collected had AIV antibodies. Based on additional testing using hemagglutination inhibition, virus neutralization, and an enzyme-linked lectin assay, 94 and 100% (2023 and 2024, respectively) of the seropositive birds tested positive for antibodies to both H5 and N1, suggesting previous infection with HPAI H5N1. These results are consistent with survival and limited clinical effects related to HPAI H5N1 infections. Furthermore, these results suggest that population immunity to HPAI H5N1 within the Chesapeake Bay might reduce future infections and potential population impacts should HP H5N1 remain on the landscape, though immunity may be waning across time. Because results are based on a single population, additional testing for both infection and antibodies as well as continued monitoring could enhance understanding of antibody persistence.

Maryland, Virginia

Highly pathogenic avian influenza virus exposure and infection in free-ranging bobcats (Lynx rufus) in New York, USA

Highly pathogenic H5N1 influenza A virus (HP H5N1) cases in wild mammals have been increasing globally. Carnivora has been the most affected mammal order; however, the extent of morbidity and mortality in carnivores exposed to HP H5N1 remains undefined. We assessed the presence of antibodies to H5 and N1 in the sera of 16 live-captured bobcats ( Lynx rufus ) in New York State, USA; reported on postcapture outcomes of HP H5N1-exposed bobcats; and reported on a case of HP H5N1 infection in one bobcat. In 2024, 4/16 bobcats (25%) tested positive for antibodies to H5 and N1. Two bobcats were confirmed alive as of November 2024 according to GPS data; the other two were alive as of June 2024, when their GPS collars stopped communicating. Another bobcat, which was negative for HP H5N1 antibodies at capture, died from HP H5N1 infection within 5 wks of capture. Our results provide evidence of bobcats both surviving and succumbing to HP H5N1 infection and highlight the importance of focused health studies paired with monitoring data to better understand exposure, infection, and outcomes for novel pathogens and species.

New York

Observational, virological, and serological data provide insights into an outbreak of highly pathogenic avian influenza among wild birds on the Yukon-Kuskokwim Delta, Alaska in 2022

In 2021–22, clade 2.3.4.4b highly pathogenic avian influenza (HPAI) viruses were introduced by wild birds into North America, leading to geographically widespread disease. In response to HPAI outbreaks throughout late 2021 and early 2022, we recorded observations of sick and dead birds, estimated abundance of carcasses, collected swab and sera samples to detect viruses, and monitored bird nesting on the Yukon-Kuskokwim Delta region of Alaska to document potential effects of disease. Thirty-six reports of sick and dead birds were registered across the region. Nineteen carcasses were opportunistically collected for diagnostic testing, of which 12 were confirmed to be infected with clade 2.3.4.4b HPAI viruses. Carcass abundance estimates from line-distance sampling provided evidence that the most common species of dead birds from the western Yukon-Kuskokwim Delta region were Cackling Goose ( Branta hutchinsii minima ), Glaucous Gull ( Larus hyperboreus ), and Black Brant ( Branta bernicla nigricans ). Only one paired cloacal and oropharyngeal swab sample from a Northern Pintail ( Anas acuta ) tested positive for clade 2.3.4.4b HPAI virus, out of 464 live-captured duck and goose samples. Of 195 sera samples from waterfowl screened for antibodies reactive to influenza A viruses, antibodies were found in 41–98% of samples collected from Emperor Goose ( Anser canagicus ), Cackling Goose, Black Brant, and Spectacled Eider ( Somateria fischeri ). In addition, 15–98% of the same sera samples were reactive to a clade 2.3.4.4b H5 antigen. Fewer Black Brant and Emperor Goose nests were found on long-term study plots during 2022 than in previous years. Collectively, we found that HPAI viruses affected at least seven species of wild birds inhabiting the region during 2022. The full scope of impacts of HPAI at this location during 2022 is unknown, but our data indicate that acute effects to avian population health on the Yukon-Kuskokwim Delta region were likely modest.

Alaska

Sampling for disease surveillance: Assessing effects on blue-winged teal survival and recovery

Outbreaks of highly pathogenic avian influenza virus in wild animals highlight the need for disease surveillance in wild birds to improve our understanding of their role as reservoirs and dispersers, and potential threats to domestic poultry and wild bird populations. Surveillance for avian influenza varies in its approach, objectives, and coordination with other monitoring efforts. For waterfowl, a common host to avian influenza viruses, banding represents a concerted effort of capturing and marking thousands of individuals annually to estimate survival and harvest rates, but users of these data have generally taken a conservative approach to remove any banded birds from analyses that had a sample taken for disease surveillance during capture. We tested for differences in survival and encounter probabilities of blue-winged teal ( Spatula discors ) marked ( n = 21,702 teal) and sampled for disease surveillance ( n = 4,216) during the nonbreeding season in Louisiana, USA, from 2016 to 2023. Although we found no consistent effect of collecting biological samples on survival probability, including an additional test showing no detectable effects of sampling for disease surveillance with oropharyngeal and cloacal swabs versus sampling with swabs and a syringe-drawn blood sample, wide 95% credible intervals on the posterior survival estimates (mean 0.36 difference between upper and lower values across all year-sex-sampling groups; 0.44 for sampling type groups) indicated low statistical power to detect an effect. Seber recovery probability during the first interval following sampling was lower among birds sampled using swabs only, but we assume this stems from low sample sizes rather than an effect of collecting biological samples. Because recovery probabilities can vary as a function of individual covariates, we also examined direct recovery probabilities and observed no meaningful effect of disease surveillance sampling type but strong effects of capture date, suggesting the effect on Seber recovery probability may have been due to heterogeneity in exposure to natural and harvest mortality risks. Although we suggest that aligning disease surveillance sample collection efforts with landscape-scale waterfowl banding efforts may have little effect on observed demographic rates, additional studies with larger sample sizes are likely needed to provide the statistical power necessary to formally conclude no effect of biological sampling on survival probabilities.

Louisiana

Genomic characterization of highly pathogenic H5 avian influenza viruses from Alaska during 2022 provides evidence for genotype-specific trends of spatiotemporal and interspecies dissemination

The ongoing panzootic of highly pathogenic H5 clade 2.3.4.4b avian influenza (HPAI) spread to North America in late 2021, with detections of HPAI viruses in Alaska beginning in April 2022. HPAI viruses have since spread across the state, affecting many species of wild birds as well as domestic poultry and wild mammals. To better understand the dissemination of HPAI viruses spatiotemporally and among hosts in Alaska and adjacent regions, we compared the genomes of 177 confirmed HPAI viruses detected in Alaska during April – December 2022. Results suggest multiple viral introductions into Alaska between November 2021 and August or September 2022, as well as dissemination to areas within and outside of the state. Viral genotypes differed in their spatiotemporal spread, likely influenced by timing of introductions relative to population immunity. We found evidence for dissemination of HPAI viruses between wild bird species, wild birds and domestic poultry, as well as wild birds and wild mammals. Continued monitoring for and genomic characterization of HPAI viruses in Alaska can improve our understanding of the evolution and dispersal of these economically costly and ecologically relevant pathogens.

Emerging Microbes & Infections

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.

Minnesota

Molecular detection and characterization of highly pathogenic H5N1 clade 2.3.4.4b avian influenza viruses among hunter-harvested wild birds provides evidence for three independent introductions into Alaska

We detected and characterized highly pathogenic avian influenza viruses among hunter-harvested wild waterfowl inhabiting western Alaska during September–October 2022 using a molecular sequencing pipeline applied to RNA extracts derived directly from original swab samples. Genomic characterization of 10 H5 clade 2.3.4.4b avian influenza viruses detected with high confidence provided evidence for three independent viral introductions into Alaska. Our results highlight the utility and some potential limits of applying molecular processing approaches directly to RNA extracts from original swab samples for viral research and monitoring.

Alasak

Demographic risk factors vary in the invasion front of chronic wasting disease in West Virginia, USA

After detecting chronic wasting disease (CWD) in white-tailed deer ( Odocoileus virginianus ) in Hampshire County, West Virginia, USA, in 2005, we investigated the change of CWD apparent prevalence and potential factors influencing infection risk during the invasion front. Over eight sampling years (2006–2012 and 2017) during a 12-yr period within a 101-km 2 -area monitoring zone, we sampled and tested a total of 853 deer for CWD by ELISA and immunohistochemistry. Bayesian logistic regression of risk factors included collection year, age class, sex, and adjusted body weight (weight after accounting for sex, age, kidney fat index, and number of fetuses). In the whole-herd model ( n =634), collection year, age, and adjusted body weight were associated with increased odds of CWD, whereas an age-weight interaction had a negative relationship. We found that males drove the positive associations with age and adjusted body weight, whereas females were responsible for the negative interaction effect. These findings suggest potential behavioral and physiological mechanisms related to sex that may influence CWD exposure. Older males exhibited higher CWD prevalence, aligning with previous studies. Notably, the novel finding of adjusted body weight as a risk factor in males warrants further investigation, and this study highlights the need for future research on social behavior and its role in CWD transmission within white-tailed deer populations.

Journal of Wildlife Diseases

Antibodies to influenza A virus in Lesser (Aythya affinis) and Greater Scaup (Aythya marila) in the USA

Scaup, including both Lesser and Greater ( Aythya affinis and Aythya marila , respectively), are a grouping of populous and widespread North American diving ducks. Few influenza type A viruses (IAV) have been reported from these species despite a high prevalence of antibodies to IAV being reported. Existing virologic and serologic data indicate that IAV infection routinely occurs in scaup, yet it is unknown which IAV subtypes are linked to these infections. In this study, we aimed to gain a more complete picture of IAV natural history in Lesser and Greater Scaup from two coastal flyways in North America in 2015–18 (302 samples from California in the Pacific Flyway and 471 samples from Maryland in the Atlantic Flyway). Low prevalence of active IAV infection was detected by real-time reverse-transcription PCR in Lesser Scaup sampled in Maryland and California (2.8% and 8.1%, respectively). A single IAV (H1N1) was isolated in embryonated chicken eggs from a bird sampled in California. Similarly low levels were observed in Greater Scaup in California (3.3%). Antibodies to the nucleoprotein as detected with a commercial blocking ELISA were observed in all species and flyway combinations. Antibody seroprevalence estimates were higher in adult Lesser Scaup than in juveniles at both the ≤0.5 ( P <0.001, z=–3.582) and ≤0.7 serum-sample-to-negative-control absorbance thresholds ( P =0.003, z=–2.996). Neutralizing antibodies to H1–H12, H14, and H15 were detected using a microtiter virus neutralization assay, with the highest prevalence of antibodies against H1 (38%), H6 (36%), and H11 (35%). The high prevalence of antibodies to IAV and evidence of previous exposure to numerous subtypes are consistent with a high level of population immunity and a low prevalence of infection. These results must be interpreted in the context of season (winter sampling), as results may vary with the annual influx of naïve juvenile birds.

California, Maryland

H5N1 highly pathogenic avian influenza clade 2.3.4.4b in wild and domestic birds: Introductions into the United States and reassortments, December 2021–April 2022

Highly pathogenic avian influenza viruses (HPAIVs) of the A/goose/Guangdong/1/1996 lineage H5 clade 2.3.4.4b continue to have a devastating effect on domestic and wild birds. Full genome sequence analyses using 1369 H5N1 HPAIVs detected in the United States (U.S.) in wild birds, commercial poultry, and backyard flocks from December 2021 to April 2022, showed three phylogenetically distinct H5N1 virus introductions in the U.S. by wild birds. Unreassorted Eurasian genotypes A1 and A2 entered the Northeast Atlantic states, whereas a genetically distinct A3 genotype was detected in Alaska. The A1 genotype spread westward via wild bird migration and reassorted with North American wild bird avian influenza viruses. Reassortments of up to five internal genes generated a total of 21 distinct clusters; of these, six genotypes represented 92% of the HPAIVs examined. By phylodynamic analyses, most detections in domestic birds were shown to be point-source transmissions from wild birds, with limited farm-to-farm spread.

Virology

Spatiotemporal changes in influenza A virus prevalence among wild waterfowl inhabiting the continental United States throughout the annual cycle

Avian influenza viruses can pose serious risks to agricultural production, human health, and wildlife. An understanding of viruses in wild reservoir species across time and space is important to informing surveillance programs, risk models, and potential population impacts for vulnerable species. Although it is recognized that influenza A virus prevalence peaks in reservoir waterfowl in late summer through autumn, temporal and spatial variation across species has not been fully characterized. We combined two large influenza databases for North America and applied spatiotemporal models to explore patterns in prevalence throughout the annual cycle and across the continental United States for 30 waterfowl species. Peaks in prevalence in late summer through autumn were pronounced for dabbling ducks in the genera Anas and Spatula , but not Mareca . Spatially, areas of high prevalence appeared to be related to regional duck density, with highest predicted prevalence found across the upper Midwest during early fall, though further study is needed. We documented elevated prevalence in late winter and early spring, particularly in the Mississippi Alluvial Valley. Our results suggest that spatiotemporal variation in prevalence outside autumn staging areas may also represent a dynamic parameter to be considered in IAV ecology and associated risks.

Scientific Reports

A lesser scaup (Aythya affinis ) naturally infected with Eurasian 2.3.4.4 highly pathogenic H5N1 avian influenza virus – Movement ecology and host factors

Despite the recognized role of wild waterfowl in the potential dispersal and transmission of highly pathogenic avian influenza (HPAI) virus, little is known about how infection affects these birds. This lack of information limits our ability to estimate viral spread in the event of an HPAI outbreak, thereby limiting our abilities to estimate and communicate risk. Here we present telemetry data from a wild Lesser Scaup ( Aythya affinis ), captured during a separate ecology study in the Chesapeake Bay, Maryland. This bird tested positive for infection with clade 2.3.4.4 HPAI virus of the A/goose/Guangdong/1/1996 (Gs/GD) H5N1 lineage (results received post-release) during the 2021–22 ongoing outbreaks in North America. While the infected bird was somewhat lighter than other adult males surgically implanted with transmitters (790g, ߂ = 868g, n = 11), it showed no clinical signs of infection at capture, during surgery, nor upon release. The bird died 3d later, pathology undetermined as the specimen was not able to be recovered. Analysis of movement data within the 3d window showed that the infected individual's maximum and average hourly movements (3894.3m, 428.8m respectively) were noticeably lower than noninfected conspecifics tagged and released the same day (߂ = 21594.5m, ߂ = 1097.9m, respectively; n = 4). We identified four instances where the infected bird had close contact (fixes located within 25m and 15 min) with another marked bird during this time. Collectively, these data suggest that the HPAI positive bird observed in this study may have been shedding virus for some period prior to death, with opportunities for direct bird to bird or environmental transmission. Although limited by low sample size and proximity to the time of tagging, we hope that these data will provide useful information as managers continue to respond to this ongoing outbreak event.

Transboundary and Emerging Diseases

Maintenance and dissemination of avian-origin influenza A virus within the northern Atlantic Flyway of North America

Wild waterbirds, the natural reservoirs for avian influenza viruses, undergo migratory movements each year, connecting breeding and wintering grounds within broad corridors known as flyways. In a continental or global view, the study of virus movements within and across flyways is important to understanding virus diversity, evolution, and movement. From 2015 to 2017, we sampled waterfowl from breeding (Maine) and wintering (Maryland) areas within the Atlantic Flyway (AF) along the east coast of North America to investigate the spatio-temporal trends in persistence and spread of influenza A viruses (IAV). We isolated 109 IAVs from 1,821 cloacal / oropharyngeal samples targeting mallards (Anas platyrhynchos) and American black ducks (Anas rubripes) , two species having ecological and conservation importance in the flyway that are also host reservoirs of IAV. Isolates with >99% nucleotide similarity at all gene segments were found between eight pairs of birds in the northern site across years, indicating some degree of stability among genome constellations and the possibility of environmental persistence. No movement of whole genome constellations were identified between the two parts of the flyway, however, virus gene flow between the northern and southern study locations was evident. Examination of banding records indicate direct migratory waterfowl movements between the two locations within an annual season, providing a mechanism for the inferred viral gene flow. Bayesian phylogenetic analyses provided evidence for virus dissemination from other North American wild birds to AF dabbling ducks (Anatinae), shorebirds (Charidriformes), and poultry (Galliformes). Evidence was found for virus dissemination from shorebirds to gulls (Laridae), and dabbling ducks to shorebirds and poultry. The findings from this study contribute to the understanding of IAV ecology in waterfowl within the AF.

Chesapeake Bay