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USGS2018

Building a response network to investigate potential pathogens associated with unionid mortality events

Unexplained mortality events have confounded the mussel conservation community for over 30 years. While the effects of chemical pollutants and environmental factors have been examined, few investigations have focused on the identification of potential mussel pathogens. Consequently, very little is known regarding the impact that pathogens have on freshwater mussel health, especially as they pertain to propagation efforts. There are several barriers that have hindered progress in the study of freshwater mussel disease, one of which is lack of appropriate diagnostic tools, and another is lack of guidance for field personnel regarding sampling procedures and laboratory contacts.

Ellipsaria↗

USGS2011

Spatial distribution and risk factors of highly pathogenic avian influenza (HPAI) H5N1 in China

Highly pathogenic avian influenza (HPAI) H5N1 was first encountered in 1996 in Guangdong province (China) and started spreading throughout Asia and the western Palearctic in 2004–2006. Compared to several other countries where the HPAI H5N1 distribution has been studied in some detail, little is known about the environmental correlates of the HPAI H5N1 distribution in China. HPAI H5N1 clinical disease outbreaks, and HPAI virus (HPAIV) H5N1 isolated from active risk-based surveillance sampling of domestic poultry (referred to as HPAIV H5N1 surveillance positives in this manuscript) were modeled separately using seven risk variables: chicken, domestic waterfowl population density, proportion of land covered by rice or surface water, cropping intensity, elevation, and human population density. We used bootstrapped logistic regression and boosted regression trees (BRT) with cross-validation to identify the weight of each variable, to assess the predictive power of the models, and to map the distribution of HPAI H5N1 risk. HPAI H5N1 clinical disease outbreak occurrence in domestic poultry was mainly associated with chicken density, human population density, and elevation. In contrast, HPAIV H5N1 infection identified by risk-based surveillance was associated with domestic waterfowl density, human population density, and the proportion of land covered by surface water. Both models had a high explanatory power (mean AUC ranging from 0.864 to 0.967). The map of HPAIV H5N1 risk distribution based on active surveillance data emphasized areas south of the Yangtze River, while the distribution of reported outbreak risk extended further North, where the density of poultry and humans is higher. We quantified the statistical association between HPAI H5N1 outbreak, HPAIV distribution and post-vaccination levels of seropositivity (percentage of effective post-vaccination seroconversion in vaccinated birds) and found that provinces with either outbreaks or HPAIV H5N1 surveillance positives in 2007–2009 appeared to have had lower antibody response to vaccination. The distribution of HPAI H5N1 risk in China appears more limited geographically than previously assessed, offering prospects for better targeted surveillance and control interventions.

PLoS Pathogens↗

USGS2025

Increased mortality rates caused by highly pathogenic avian influenza virus in a migratory raptor

Highly pathogenic avian influenza virus (HPAIV) has caused extensive mortalities in wild birds with a disproportionate impact on raptors since 2021. The population-level impact of HPAIV can be informed by telemetry studies that track large samples of initially healthy, wild birds. We leveraged movement data from 71 rough-legged hawks ( Buteo lagopus ) across all major North American migratory bird flyways concurrent with the 2022–2023 HPAIV outbreak and identified a total of 29 mortalities, of which 11 were confirmed, and an additional ~9 were estimated to have been caused by HPAIV. We estimated a 28% HPAIV cause-specific mortality rate among rough-legged hawks during a single year concurrent with the HPAIV outbreak in North America. Additionally, the overall mortality rate during the HPAIV outbreak (47%) was significantly higher than baseline annual mortality rates (3%–17%) suggesting that HPAIV-caused deaths were additive above baseline mortality levels. HPAIV mortalities were concentrated within the Central and Atlantic flyways during prebreeding migration and peaked in April 2022 when large-scale HPAIV mortalities were reported in other wild birds throughout North America. HPAIV exposure was most likely caused by scavenging or preying on infected waterfowl, as rough-legged hawks are known to opportunistically scavenge during the nonbreeding season. We utilized movement data to identify a continental-scale HPAIV cause-specific mortality event in rough-legged hawks that has the potential to exacerbate ongoing population declines. Our study highlights the usefulness of monitoring movement data to pinpoint sources of mortality that can help better understand the drivers of population change, even if studies are focused on other research questions.

Ecology and Evolution↗

USGS2021

Natural history of a bighorn sheep pneumonia epizootic: Source of infection, course of disease, and pathogen clearance

A respiratory disease epizootic at the National Bison Range (NBR) in Montana in 2016–2017 caused an 85% decline in the bighorn sheep population, documented by observations of its unmarked but individually identifiable members, the subjects of an ongoing long-term study. The index case was likely one of a small group of young bighorn sheep on a short-term exploratory foray in early summer of 2016. Disease subsequently spread through the population, with peak mortality in September and October and continuing signs of respiratory disease and sporadic mortality of all age classes through early July 2017. Body condition scores and clinical signs suggested that the disease affected ewe groups before rams, although by the end of the epizootic, ram mortality (90% of 71) exceeded ewe mortality (79% of 84). Microbiological sampling 10 years to 3 months prior to the epizootic had documented no evidence of infection or exposure to Mycoplasma ovipneumoniae at NBR, but during the epizootic, a single genetic strain of M. ovipneumoniae was detected in affected animals. Retrospective screening of domestic sheep flocks near the NBR identified the same genetic strain in one flock, presumptively the source of the epizootic infection. Evidence of fatal lamb pneumonia was observed during the first two lambing seasons following the epizootic but was absent during the third season following the death of the last identified M. ovipneumoniae carrier ewe. Monitoring of life-history traits prior to the epizootic provided no evidence that environmentally and/or demographically induced nutritional or other stress contributed to the epizootic. Furthermore, the epizootic occurred despite proactive management actions undertaken to reduce risk of disease and increase resilience in this population. This closely observed bighorn sheep epizootic uniquely illustrates the natural history of the disease including the (presumptive) source of spillover, course, severity, and eventual pathogen clearance.

Montana↗

USGS2026

Cost of migration increased during an outbreak of highly pathogenic avian influenza virus

Migration is thought to be costly such that challenges faced during migration likely affect how birds migrate through direct selection on migratory behavior. Survival throughout the annual cycle and conditions that affect the survival costs of migration are therefore critical to our understanding of the causes and consequences of migration. Estimating daily survival throughout the annual cycle of migratory birds has only recently become possible. We used over a decade of GPS tracking data from 186 migratory Buteo lagopus to quantify causes of mortality as well as demographic, seasonal, and spatial variation in daily survival and how survival consequences of migration changed during an outbreak of highly pathogenic avian influenza virus. The cost of migration via direct mortality varied between avian influenza and baseline study periods. Surprisingly, mortality risk during typical spring migrations was lower than in all other seasons, but mortality risk during the avian influenza outbreak was higher during both spring and fall migrations. Additionally, we found increased mortality risk in females during the avian influenza outbreak but no sex differences during baseline periods. Hence, the relative cost of migration was altered by the virus. During baseline periods, mortality risk was highest during late fall migration and winter, coinciding with increased anthropogenic-related mortalities at lower latitudes. However, the number of deaths caused by avian influenza (n = 11) in a single year equaled the total number of deaths caused by anthropogenic-related factors (n = 11; primarily collision trauma and persecution) across all 12 study years. Overall, our study highlights how added stressors such as disease outbreaks and anthropogenic factors may exacerbate existing seasonal constraints by directly increasing mortality risk.

Journal of Avian Biology↗

USGS2026

Quantitative microbial risk assessment with microbial source tracking for enteric pathogens in southwest Wisconsin private wells

Private wells supply drinking water for many households, and their contamination by fecal microbes presents a risk of acute gastrointestinal illness (AGI). Risk is thought to vary by contaminating fecal source, but specific associations with fecal source are unknown for most private wells in the United States. This study characterizes AGI risk in Grant, Iowa, and Lafayette counties in southwestern Wisconsin. AGI risk was assessed for 10 viral, bacterial, and protozoan pathogens detected in private wells using quantitative microbial risk assessment. Exposure assessment was based on sampling 138 private wells in the study area, and risk was associated with fecal source by microbial source tracking (MST). Median Monte Carlo estimates indicated 4450 AGI cases/year in the study area associated with drinking water from contaminated private wells (95% confidence interval: 90–37,990). Most annual cases were associated with detection of human MST markers, including median estimates of 2550 associated with only human markers and 880 with human and livestock MST markers detected together. Note that 50 AGI cases/year were associated with detection of only livestock markers, and nearly 1000 cases were estimated to occur in the absence of detectable MST markers. This study characterizes AGI risk for households served by private wells in southwestern Wisconsin. It illustrates that human fecal sources can predominate risk associated with drinking water from private wells, even in a rural region characterized by substantial agricultural activity. MST can characterize the relationship between risk and fecal source; contributions of fecal sources to risk cannot be assumed from land use.

Wisconsin↗

USGS2016

Concordance in diagnostic testing for respiratory pathogens of bighorn sheep

Reliable diagnostic tests are essential for disease investigation and management. This is particularly true for diseases of free-ranging wildlife where sampling is logistically difficult precluding retesting. Clinical assays for wildlife diseases frequently vary among laboratories because of lack of appropriate standardized commercial kits. Results of diagnostic testing may also be called into question when investigators report different etiologies for disease outbreaks, despite similar clinical and pathologic findings. To evaluate reliability of diagnostic testing for respiratory pathogens of bighorn sheep ( Ovis canadensis ), we conducted a series of ring tests across 6 laboratories routinely involved in detection of Mycoplasma ovipneumoniae , Pasteurellaceae, lktA (the Pasteurellaceae gene encoding leukotoxin), and 3 reference laboratories. Consistency of results for replicate samples within laboratories was high (median agreement = 1.0). Agreement between laboratories was high for polymerase chain reaction (PCR) detection of M. ovipneumoniae and culture isolation of Mannheimia spp. and Bibersteinia trehalosi (median agreement = 0.89–0.95, Kappa = 0.65–0.74), and lower for PCR detection of Mannheimia spp. lktA (median agreement = 0.58, Kappa = 0.12). Most errors on defined status samples were false negatives, suggesting test sensitivity was a greater problem than specificity. However, tests for M. haemolytica and lktA yielded some false positive results. Despite differences in testing protocols, median agreement among laboratories and correct classification of controls for most agents was ≥0.80, meeting or exceeding the standard required by federal proficiency testing programs. This information is valuable for interpreting test results, laboratory quality assessments, and advancing diagnosis of respiratory disease in wild sheep. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

Wildlife Society Bulletin↗

USGS1997

Characterization of a linear DNA plasmid from the filamentous fungal plant pathogen Glomerella musae [Anamorph: Colletotrichum musae (Berk. and Curt.) arx.]

A 7.4-kilobase (kb) DNA plasmid was isolated from Glomerella musae isolate 927 and designated pGML1. Exonuclease treatments indicated that pGML1 was a linear plasmid with blocked 5' termini. Cell-fractionation experiments combined with sequence-specific PCR amplification revealed that pGML1 resided in mitochondria. The pGML1 plasmid hybridized to cesium chloride-fractionated nuclear DNA but not to A + T-rich mitochondrial DNA. An internal 7.0-kb section of pGML1 was cloned and did not hybridize with either nuclear or mitochondrial DNA from G. musae. Sequence analysis revealed identical terminal inverted repeats (TIR) of 520 bp at the ends of the cloned 7.0-kb section of pGML1. The occurrence of pGML1 did not correspond with the pathogenicity of G. musae on banana fruit. Four additional isolates of G. musae possessed extrachromosomal DNA fragments similar in size and sequence to pGML1.

Current Genetics↗

USGS2010

A reverse genetics system for the Great Lakes strain of viral hemorrhagic septicemia virus: the NV gene is required for pathogenicity

Viral hemorrhagic septicemia virus (VHSV), belonging to the genus Novirhabdovirus in the family of Rhabdoviridae , causes a highly contagious disease of fresh and saltwater fish worldwide. Recently, a novel genotype of VHSV, designated IVb, has invaded the Great Lakes in North America, causing large-scale epidemics in wild fish. An efficient reverse genetics system was developed to generate a recombinant VHSV of genotype IVb from cloned cDNA. The recombinant VHSV (rVHSV) was comparable to the parental wild-type strain both in vitro and in vivo, causing high mortality in yellow perch ( Perca flavescens ). A modified recombinant VHSV was generated in which the NV gene was substituted with an enhanced green fluorescent protein gene (rVHSV-ΔNV-EGFP), and another recombinant was made by inserting the EGFP gene into the full-length viral clone between the P and M genes (rVHSV-EGFP). The in vitro replication kinetics of rVHSV-EGFP was similar to rVHSV; however, the rVHSV-ΔNV-EGFP grew 2 logs lower. In yellow perch challenges, wtVHSV and rVHSV induced 82-100% cumulative per cent mortality (CPM), respectively, whereas rVHSV-EGFP produced 62% CPM and rVHSV-ΔNV-EGFP caused only 15% CPM. No reversion of mutation was detected in the recovered viruses and the recombinant viruses stably maintained the foreign gene after several passages. These results indicate that the NV gene of VHSV is not essential for viral replication in vitro and in vivo, but it plays an important role in viral replication efficiency and pathogenicity. This system will facilitate studies of VHSV replication, virulence, and production of viral vectored vaccines.

Marine Biotechnology↗

USGS2011

Flying over an infected landscape: Distribution of highly pathogenic avian influenza H5N1 risk in South Asia and satellite tracking of wild waterfowl

Highly pathogenic avian influenza (HPAI) H5N1 virus persists in Asia, posing a threat to poultry, wild birds, and humans. Previous work in Southeast Asia demonstrated that HPAI H5N1 risk is related to domestic ducks and people. Other studies discussed the role of migratory birds in the long distance spread of HPAI H5N1. However, the interplay between local persistence and long-distance dispersal has never been studied. We expand previous geospatial risk analysis to include South and Southeast Asia, and integrate the analysis with migration data of satellite-tracked wild waterfowl along the Central Asia flyway. We find that the population of domestic duck is the main factor delineating areas at risk of HPAI H5N1 spread in domestic poultry in South Asia, and that other risk factors, such as human population and chicken density, are associated with HPAI H5N1 risk within those areas. We also find that satellite tracked birds (Ruddy Shelduck and two Bar-headed Geese) reveal a direct spatio-temporal link between the HPAI H5N1 hot-spots identified in India and Bangladesh through our risk model, and the wild bird outbreaks in May,June,July 2009 in China(Qinghai Lake), Mongolia, and Russia. This suggests that the continental-scale dynamics of HPAI H5N1 are structured as a number of persistence areas delineated by domestic ducks, connected by rare transmission through migratory waterfowl.

EcoHealth↗

USGS2019

Overview of emerging amphibian pathogens and modeling advances for conservation-related decisions

One of the leading causes of global amphibian decline is emerging infectious disease. We summarize the disease ecology of four major emerging amphibian infectious agents: chytrids, ranaviruses, trematodes, and Perkinsea. We focus on recently developed quantitative advances that build on well-established ecological theories and aid in studying epizootic and enzootic disease dynamics. For example, we identify ecological and evolutionary selective forces that determine disease outcomes and transmission pathways by borrowing ideas from population and community ecology theory. We outline three topics of general interest in disease ecology: (i) the relationship between biodiversity and disease risk, (ii) individual, species, or environmental transmission heterogeneity, and (iii) pathogen coinfections. Finally, we identify specific knowledge gaps impeding the success of conservation-related decisions for disease mitigation and the future of amphibian conservation success.

Biological Conservation↗

USGS2021

Evaluation of regulatory action and surveillance as preventive risk-mitigation to an emerging global amphibian pathogen Batrachochytrium salamandrivorans (Bsal)

The emerging amphibian pathogen Batrachochytrium salamandrivorans (Bsal) is a severe threat to global urodelan (salamanders, newts, and related taxa) biodiversity. Bsal has not been detected, to date, in North America, but the risk is high because North America is one of the global hotspots for urodelan biodiversity. The North American and United States response to the discovery of Bsal in Europe was to take a risk-based approach to preventive management actions, including interim regulations on importation of captive salamanders and a large-scale surveillance effort. Risk-based approaches to decision-making can extend to adaptive management cycles by periodically incorporating new information that reduces uncertainty in an estimate of risk or to assess the effect of mitigation actions which reduce risk directly. Our objectives were to evaluate the effects of regulatory action on the introduction of Bsal to the U.S., quantify how a large-scale surveillance effort impacted consequence risk, and to combine other new information on species susceptibility to re-evaluate Bsal risk to the U.S. Import regulations effectively reduced import volume of targeted species, but new research on species susceptibility suggests the list of regulated species was incomplete regarding Bsal reservoir species. Not detecting Bsal in an intensive surveillance effort improved confidence that Bsal was not present, however, the overall risk-reduction impact was limited because of the expansive area of interest (conterminous United States) and limited time frame of sampling. Overall, the preventive actions in response to the Bsal threat did reduce Bsal risk in the U.S. and we present an updated risk assessment to provide information for adaptive decision-making.

Biological Conservation↗

USGS2023

The changing dynamics of highly pathogenic avian influenza H5N1: Next steps for management & science in North America

Highly pathogenic avian influenza virus (HPAIV) H5N1 was introduced in North America in late 2021 through trans-Atlantic pathways via migratory birds. These introductions have resulted in an unprecedented epizootic, a widespread disease event in animals, heavily affecting poultry, wild birds, and recently mammals. The North American incursions occurred during the largest epidemic season (2021–2022) in Europe where H5N1 may now be endemic (i.e., continuously present). The continuing outbreak includes expansion into Mexico, Central and South America beginning in late 2022. Here, we provide an overview of the Eurasian origin H5N1 introduction to the Americas, including a significant shift in virus dynamics and severe disease in wild birds. Then, to investigate the global changes in confirmed detections in wild birds and poultry across time and geographic regions, we analyzed FAO's EMPRES-i + database. To examine the 2021 introduction and spread in North American wild birds and poultry, we collated publicly available data across USA and Canadian federal sources. Based on our assessment, the unique magnitude of the North American H5N1 spread indicates the need for effective decision framing to prioritize management needs and scientific inquiry, particularly for species at risk and interface areas for wildlife, poultry, and humans. We illustrate the rapidly occurring and likely increasing detrimental effects that this One Health issue has on wildlife, agriculture, and potentially human health, and we offer a reframing of HPAIV disease response towards a decision analytical context to guide scientific prioritization as a potentially valuable change in focus.

Biological Conservation↗

USGS2013

Bactericidal efficacy of elevated pH on fish pathogenic and environmental bacteria

Ship ballast water is a recognized medium for transfer and introductions of nonindigenous species. There is a need for new ballast water treatment methods that effectively and safely eliminate or greatly minimize movements of these species. The present study employed laboratory methods to evaluate the bactericidal efficacy of increased pH (pH 10.0–12.0) for exposure durations of up to 72 h to kill a variety of Gram-negative and Gram-positive bacteria including fish pathogens (Aeromonas spp., Yersinia ruckeri, Edwardsiella ictaluri, Serratia liquefaciens, Carnobacterium sp.), other common aquatic-inhabitant bacteria (Serratia marcescens, Pseudomonas fluorescens, Staphylococcus sp., Bacillus sp.) and indicators listed in International Maritime Organization D2 Standards; namely, Vibrio cholera (an environmental isolate from fish), Escherichia coli and Enterococcus faecalis. Volumes of 5 N NaOH were added to tryptic soy broth to obtain desired pH adjustments. Viable cells were determined after 0, 4, 12, 24, 48, and 72 h. Initial (0 h) cell numbers ranged from 3.40 × 10 4 cfu/mL for Bacillus sp. to 2.44 × 10 7 cfu/mL for E. faecalis. The effective endpoints of pH and treatment duration necessary to realize 100% bactericidal effect varied; however, all bacteria tested were killed within 72 h at pH 12.0 or lower. The lowest parameters examined, 4 h at pH 10.0, were bactericidal to V. cholera, E. ictaluri, three of four isolates of E. coli, and (three of four) Aeromonas salmonicida subsp. salmonicida. Bactericidal effect was attained at pH 10.0 within 12 h for the other A. salmonicida subsp. salmonicida, and within 24 h for P. fluorescens, and the remaining E. coli.

Journal of Advanced Research↗

USGS2011

Interspecies transmission and limited persistence of low pathogenic avian influenza genomes among Alaska dabbling ducks

The reassortment and geographic distribution of low pathogenic avian influenza (LPAI) virus genes are well documented, but little is known about the persistence of intact LPAI genomes among species and locations. To examine persistence of entire LPAI genome constellations in Alaska, we calculated the genetic identities among 161 full-genome LPAI viruses isolated across 4 years from five species of duck: northern pintail ( Anas acuta ), mallard ( Anas platyrhynchos ), American green-winged teal ( Anas crecca ), northern shoveler ( Anas clypeata ) and American wigeon ( Anas americana ). Based on pairwise genetic distance, highly similar LPAI genomes (>99% identity) were observed within and between species and across a range of geographic distances (up to and >1000 km), but most often between isolates collected 0–10 km apart. Highly similar viruses were detected between years, suggesting inter-annual persistence, but these were rare in our data set with the majority occurring within 0–9 days of sampling. These results identify LPAI transmission pathways in the context of species, space and time, an initial perspective into the extent of regional virus distribution and persistence, and insight into why no completely Eurasian genomes have ever been detected in Alaska. Such information will be useful in forecasting the movement of foreign-origin avian influenza strains should they be introduced to North America.

Alaska↗

USGS2020

Modeling pathogen dispersal in marine fish and shellfish

Bio-physical models are a useful tool for understanding dispersal and transmission of marine pathogens. While utilized for larval dispersal models, they are only recently being used in epidemiological studies and are currently underutilized by the marine epidemiology field. Bio-physical models are useful for spatial planning and coastal management. For example, they have been used for spatial planning of salmon farm site locations, and to establish early warning networks. Bio-physical modeling can be used to test hypotheses, rather than simply develop them. Model resolution and computation demands must be balanced when making decisions about model parameters. Epidemiological bio-physical models are in their infancy. While they have proven useful so far, future applications of these models can incorporate more aspects of disease dynamics and address many additional questions.

Trends in Parasitology↗

USGS2014

Humic substances interfere with detection of pathogenic prion protein

Studies examining the persistence of prions (the etiological agent of transmissible spongiform encephalopathies) in soil require accurate quantification of pathogenic prion protein (PrP TSE ) extracted from or in the presence of soil particles. Here, we demonstrate that natural organic matter (NOM) in soil impacts PrP TSE detection by immunoblotting. Methods commonly used to extract PrPTSE from soils release substantial amounts of NOM, and NOM inhibited PrPTSE immunoblot signal. The degree of immunoblot interference increased with increasing NOM concentration and decreasing NOM polarity. Humic substances affected immunoblot detection of prion protein from both deer and hamsters. We also establish that after interaction with humic acid, PrP TSE remains infectious to hamsters inoculated intracerebrally, and humic acid appeared to slow disease progression. These results provide evidence for interactions between PrPTSE and humic substances that influence both accurate measurement of PrP TSE in soil and disease transmission.

Soil Biology and Biochemistry↗

USGS2010

Intercontinental reassortment and genomic variation of low pathogenic avian influenza viruses isolated from northern pintails ( Anas acuta ) in Alaska: examining the evidence through space and time

Migration and population genetic data for northern pintails ( Anas acuta ) and phylogenetic analysis of low pathogenic avian influenza (LPAI) viruses from this host in Alaska suggest that northern pintails are involved in ongoing intercontinental transmission of avian influenza. Here, we further refine this conclusion through phylogenetic analyses which demonstrate that detection of foreign lineage gene segments is spatially dependent and consistent through time. Our results show detection of foreign lineage gene segments to be most likely at sample locations on the Alaska Peninsula and least likely along the Southern Alaska Coast. Asian lineages detected at four gene segments persisted across years, suggesting maintenance in avian hosts that migrate to Alaska each year from Asia or in hosts that remain in Alaska throughout the year. Alternatively, live viruses may persist in the environment and re-infect birds in subsequent seasons.

Alaska↗