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The impacts of co-circulating pathogens in Pacific herring depend on interactions between viral life-cycle traits and transmission parameters, highlighting interdependencies between pathogen epizootics

The average host susceptibility decreases as the epizootic progresses because easily infected hosts are first removed from the population. While host susceptibility is pathogen-specific, it is likely that host susceptibility is correlated between different pathogens, so that co-circulating pathogens may have reciprocal impacts on their epidemics. However, despite well-documented examples of concomitant infections in marine hosts, reciprocal epizootic effects have not been documented in wild marine organisms. We quantify reciprocal impacts between viral haemorrhagic septicaemia and viral erythrocytic necrosis in Pacific herring ( Clupea pallasii ) using field and laboratory work. We show that the causative viruses for both diseases circulate through herring populations and that infection with one pathogen has negative impacts on the epizootic and infection characteristics of the other pathogen, suggesting positive correlations in the susceptibility to infection between pathogens. We then use simulations of a two-strain pathogen model to show that the impact of the correlation is modulated by transmission parameters, such as the incubation period and the initial transmission rate. Our work shows that co-occurring epizootics pose a management challenge because single-pathogen management actions may amplify the epizootics of the non-targeted pathogen. This study provides a framework to evaluate the consequences of reciprocal epizootic impacts through field, experimental and modelling work.

Washington

Infection–nutrition feedbacks: Fat supports pathogen clearance but pathogens reduce fat in a wild mammal

Though far less obvious than direct effects (clinical disease or mortality), the indirect influences of pathogens are difficult to estimate but may hold fitness consequences. Here, we disentangle the directional relationships between infection and energetic reserves, evaluating the hypotheses that energetic reserves influence infection status of the host and that infection elicits costs to energetic reserves. Using repeated measures of fat reserves and infection status in individual bighorn sheep ( Ovis canadensis ) in the Greater Yellowstone Ecosystem, we documented that fat influenced ability to clear pathogens ( Mycoplasma ovipneumoniae ) and infection with respiratory pathogens was costly to fat reserves. Costs of infection approached, and in some instances exceeded, costs of rearing offspring to independence in terms of reductions to fat reserves. Fat influenced probability of clearing pathogens, pregnancy and over-winter survival; from an energetic perspective, an animal could survive for up to 23 days on the amount of fat that was lost to high levels of infection. Cost of pathogens may amplify trade-offs between reproduction and survival. In the absence of an active outbreak, the influence of resident pathogens often is overlooked. Nevertheless, the energetic burden of pathogens likely has consequences for fitness and population dynamics, especially when food resources are insufficient.

Wyoming

Host vs. pathogen evolutionary arms race: Effects of exposure history on individual response to a genetically diverse pathogen

Introduction: Throughout their range, bighorn sheep ( Ovis canadensis ) populations have seen significant disease-associated declines. Unfortunately, understanding of the underlying epidemiological processes driving the disease dynamics in this species has hindered conservation efforts aimed at improving the health and long-term viability of these populations. Individual response to pathogen exposure emerges from dynamic interactions between competing evolutionary processes within the host and pathogen. The host’s adaptive immune system recognizes pathogens and mounts a defensive response. Pathogens have evolved strategies to overcome adaptive immune defenses including maintaining high genetic diversity through rapid evolution. The outcomes of this evolutionary warfare determine the success of pathogen invasion of the host and ultimately the success of conservation efforts. Methods: During an epizootic dominated by a single strain, we explore these host-pathogen dynamics by examining the variation in effects of pathogen invasion on captive bighorn sheep with differing histories of exposure to genetically diverse strains of Mycoplasma ovipneumoniae (Movi). We monitored clinical signs of disease and sampled animals and their environment to detect spread of Movi among 37 bighorn sheep separated into nine pens based on known exposure Results: We documented Movi transmission within and across pens and we detected Movi DNA in air, water, and invertebrate samples. Higher levels of antibody to Movi prior to the epizootic were associated with a lower likelihood of presenting clinical signs of pneumonia. Nonetheless, higher antibody levels in symptomatic individuals were associated with more severe progressive disease, increased probability and speed of pneumonia-induced mortality, and reduced likelihood of returning to a healthy state. Bighorn sheep with previous exposure to a strain other than the predominant epizootic strain were more likely to recover. Discussion: Our results indicate that Movi-strain variability was sufficient to overwhelm the adaptive host immunological defenses. This outcome indicates, in free-ranging herds, past exposure is likely insufficient to protect bighorn sheep from infection by new Movi strains, although it influences the progression of disease and recovery within the herd. Therefore, given Movi-strain variability and the lack of immunological protection from past exposure, focusing management efforts on minimizing the introduction of Movi into bighorn herds, through separation of domestic and bighorn sheep and avoidance of management activities that create commingling of bighorn sheep carrying differing Movi strains, will likely be the most effective approach for reducing the effects of disease and achieving bighorn sheep conservation goals.

Idaho, Oregon, South Dakota, Washington

Variation in pathogen load and the pathogen load–infectiousness relationship broaden avian malaria’s distribution

Two aspects of host infectiousness shape pathogen transmission and distribution but are underappreciated: the relationship between pathogen load and infectiousness, and variability in pathogen load within species. We quantified the relationship between host pathogen load (parasitemia) for avian malaria ( Plasmodium relictum ) and infectiousness for biting Culex quinquefasciatus mosquitoes with experimental infections in canaries ( Serinus canaria ). Using this relationship, we estimated the infectiousness of 17 bird species in 11 communities in Hawaiʻi and quantified the relative contributions of infection stage (acute versus chronic) to transmission. We show that infectiousness to mosquitoes increased with parasitemia, temperature, and time since feeding. The relationship’s gradual (low) parasitemia slope resulted in a wide range of parasitemias being partly infectious, and high within-host species variability in parasitemia led to extensive overlap in infectiousness among hosts. Disproportionate mosquito host utilization (inferred from relative infection prevalence) elevated the importance of a few host species, yet broad overlap in species infectiousness resulted in similar total infectiousness across most bird communities. This similarity likely contributed to avian malaria’s widespread distribution throughout Hawaiʻi despite diverse host community assemblages. Our findings highlight the importance of both the shape of the pathogen load–infectiousness relationship and within-species variability in determining a pathogen’s host range, transmission intensity, and spatial spread.

Nature Communications

Development of an aquatic pathogen database (AquaPathogen X) and its utilization in tracking emerging fish virus pathogens in North America

The AquaPathogen X database is a template for recording information on individual isolates of aquatic pathogens and is freely available for download (http://wfrc.usgs.gov). This database can accommodate the nucleotide sequence data generated in molecular epidemiological studies along with the myriad of abiotic and biotic traits associated with isolates of various pathogens (e.g. viruses, parasites and bacteria) from multiple aquatic animal host species (e.g. fish, shellfish and shrimp). The cataloguing of isolates from different aquatic pathogens simultaneously is a unique feature to the AquaPathogen X database, which can be used in surveillance of emerging aquatic animal diseases and elucidation of key risk factors associated with pathogen incursions into new water systems. An application of the template database that stores the epidemiological profiles of fish virus isolates, called Fish ViroTrak, was also developed. Exported records for two aquatic rhabdovirus species emerging in North America were used in the implementation of two separate web-accessible databases: the Molecular Epidemiology of Aquatic Pathogens infectious haematopoietic necrosis virus (MEAP-IHNV) database (http://gis.nacse.org/ihnv/) released in 2006 and the MEAP- viral haemorrhagic septicaemia virus (http://gis.nacse.org/vhsv/) database released in 2010.

Journal of Fish Diseases

Low-pathogenic influenza A viruses in North American diving ducks contribute to the emergence of a novel highly pathogenic influenza A(H7N8) virus

Introductions of low-pathogenic avian influenza (LPAI) viruses of subtypes H5 and H7 into poultry from wild birds have the potential to mutate to highly pathogenic avian influenza (HPAI) viruses, but such viruses' origins are often unclear. In January 2016, a novel H7N8 HPAI virus caused an outbreak in turkeys in Indiana, USA. To determine the virus's origin, we sequenced the genomes of 441 wild-bird origin influenza A viruses (IAVs) from North America and subjected them to evolutionary analyses. The results showed that the H7N8 LPAI virus most likely circulated among diving ducks in the Mississippi flyway during autumn 2015 and was subsequently introduced to Indiana turkeys, in which it evolved high pathogenicity. Preceding the outbreak, an isolate with six gene segments (PB2, PB1, PA, HA, NA, and NS) sharing >99% sequence identity with those of H7N8 turkey isolates was recovered from a diving duck sampled in Kentucky, USA. H4N8 IAVs from other diving ducks possessed five H7N8-like gene segments (PB2, PB1, NA, MP, and NS; >98% sequence identity). Our findings suggest that viral gene constellations circulating among diving ducks can contribute to the emergence of IAVs that affect poultry. Therefore, diving ducks may serve an important and understudied role in the maintenance, diversification, and transmission of IAVs in the wild-bird reservoir.

Journal of Virology

Genetic conversion of a fungal plant pathogen to a non-pathogenic, endophytic mutualist

The filamentous fungal ascomycete Colletotrichum magna causes anthracnose in cucurbit plants. Isolation of a nonpathogenic mutant of this species (path-1) resulted in maintained wild-type levels of in vitro sporulation, spore adhesion, appressorial formation, and infection. Path-1 grew throughout host tissues as an endophyte and retained the wild-type host range, which indicates that the genetics involved in pathogenicity and host specificity are distinct. Prior infection with path-1 protected plants from disease caused by Colletotrichum and Fusarium .Genetic analysis of a cross between path-1 and wild-type strains indicated mutation of a single locus.

Science

Microbial and viral indicators of pathogens and human health risks from recreational exposure to waters impaired by fecal contamination

Fecal indicator bacteria (FIB) (e.g., fecal coliforms, Escherichia coli , and enterococci) have been used for decades to monitor for and protect the public from waterborne pathogens from fecal contamination. However, FIB may not perform well at predicting the presence of waterborne pathogens or human health outcomes from recreational exposure to fecal-contaminated surface waters. Numerous factors can influence the relationship between FIB and pathogens or human health outcomes, including the source(s) of contamination, the type of pathogen(s) present, differences in the survival and behavior of FIB and pathogens in the wastewater conveyance and treatment process, and varying environmental conditions. As a result, different indicators, such as source-specific microbial source tracking (MST) markers and viral fecal indicators, have been used as possible surrogates to better approximate pathogen abundance and human health risks in recreational waters. The performance of these alternative indicators has been mixed, with some promise of viral indicators better approximating viral pathogens than bacterial fecal indicators, and FIB generally more closely associated with bacterial and protozoal pathogen presence than human MST markers. Many of the assays to detect and quantify fecal indicators and pathogens are polymerase chain reaction-based assays, which detect and quantify nucleic acid [deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)] sequences specific to a target of interest. Recent advances in DNA and RNA sequencing technologies may push the field toward metabarcoding approaches, where multiple targets can be detected and quantified simultaneously. Metabarcoding is currently more applicable to bacterial and protozoal assessments than viral assessments based on a lack of universal metabarcoding markers for viruses. Innovative technologies, such as biosensors and nanotechnologies, may provide more sensitive and accurate tools to detect and quantify pathogens. When a specific pathogen is of concern for a recreational water body, a practical approach in estimating the likelihood of human health outcomes is the application of quantitative microbial risk assessments (QMRAs). Quantitative microbial risk assessments can be used to model the likelihood of pathogen-specific human health outcomes from recreational exposure as a function of a surrogate indicator. Inputs for QMRAs include the ratio between the indicator to be monitored and the pathogen of interest, the concentration of the indicator, the amount of water ingested, and the likelihood of the health outcome based on the estimated amount of pathogen consumed. There are numerous unknowns about the behavior and survival of fecal indicators and pathogens in environmental waters. Developing accurate models to predict pathogen concentrations from fecal indicators in recreational waters will require a better understanding of these unknowns. Current methods and technologies for detecting and quantifying fecal indicators and pathogens are limited due to the rare and patchy nature of pathogens. Technological advances may help improve sensitivity for detecting and quantifying pathogens.

Journal of Sustainable Water in the Built Environm

Evaluation of pathogen risks and testing considerations for Chinook salmon egg movements between New Zealand and California

Executive Summary Oncorhynchus tshawytscha (Walbaum in Artedi, 1792; Chinook salmon) were historically abundant in the McCloud River but are now extirpated from this tributary owing to dam construction and lack of passage. Planning efforts to restore populations above Shasta and Keswick Dams are currently underway, including an evaluation of potential source populations. One potential source is New Zealand Chinook salmon, which are believed to have originated from tributaries of the Sacramento River. These fish could be returned to California if reintroduction risks, including risks of pathogen introduction, could be sufficiently mitigated. The U.S. Geological Survey was contracted to provide scientific support for reintroduction efforts, including evaluating the risks of pathogen transmission via the movement of Chinook salmon eggs from New Zealand to the McCloud River. This report estimates pathogen risks associated with egg movement and considers epidemiological and biosecurity measures to minimize these risks. Pathogen risks associated with the movement of Chinook salmon eggs from New Zealand were evaluated based on pathogen virulence, transmission route, and geographic distribution. These criteria identified 14 moderate- and high-risk pathogens out of the 30 pathogens evaluated. Pathogen species and strains were considered high risk if they have the potential for vertical transmission (that is, transmission from parent to offspring), are moderately or highly virulent, and are exotic to the Sacramento River Basin. According to these criteria, we identified the following pathogens as high risk: New Zealand rickettsia-like organisms 1 and 2. —These bacterial pathogens have been associated with mortality events in farmed Chinook salmon from the South Island of New Zealand but have not been detected in other regions. Pilchard orthomyxovirus (POMV). —POMV has been detected in Sardina pilchardus (Walbaum, 1792; pilchards) and Salmo salar (Linnaeus, 1758; Atlantic salmon) from the coasts of southern Australia and Tasmania. POMV can cause relatively high mortality rates and may be indirectly transmitted via contaminated water sources. Infectious pancreatic necrosis virus (IPNV). —IPNV has a wide geographic distribution and is present in the Sacramento River Basin, but the IPNV-like viruses detected in Australia and New Zealand are unique from those found in the United States. Yersinia ruckeri . —This bacterial pathogen is the causative agent of enteric redmouth disease and has a widespread geographic distribution. However, the strains that are present in Australia and New Zealand are unique from those found in North America. Strategic use of testing and biosecurity measures can minimize pathogen risks associated with the movement of eggs. The most effective measures include iodophor treatment of eggs to remove external pathogens, testing of all the adult fish from which gametes are obtained, and a quarantine period after transport to confirm pathogen testing results. Additional measures to enhance biosecurity could include testing the quarantined fish following emergence and (or) developing a fish health history of the source population through pathogen monitoring.

California

A novel method for estimating pathogen presence, prevalence, load, and dynamics at multiple scales

The use of quantitative real-time PCR (qPCR) to monitor pathogens is common; however, quantitative frameworks that consider the observation process, dynamics in pathogen presence, and pathogen load are lacking. This can be problematic in the early stages of disease progression, where low level detections may be treated as ‘inconclusive’ and excluded from analyses. Alternatively, a framework that accounts for imperfect detection would provide more robust inferences. To better estimate pathogen dynamics, we developed a hierarchical multi-scale dynamic occupancy hurdle model (MS-DOHM). The model used data gathered during sampling for Pseudogymnoascus destructans (Pd) , the causative agent of white-nose syndrome, a fungal disease that has cause severe declines in several species of hibernating bats in North America. The model allowed us to estimate initial occupancy, colonization, persistence and prevalence of Pd at bat hibernacula. Additionally, utilizing the relationship between cycle threshold and pathogen load, we estimated pathogen detectability and modeled expected colony and bat pathogen loads. To assess the ability of MS-DOHM to estimate pathogen dynamics, we compared MS-DOHM’s results to those of a dynamic occupancy model and naïve detection/non-detection. MS-DOHM’s estimates of site-level pathogen presence were up to 11.9% higher than estimates from the dynamic occupancy model and 35.7% higher than naïve occupancy. Including prevalence and load in our modeling framework resulted in estimates of pathogen arrival that were two to three years earlier compared to the dynamic occupancy and naïve detection/non-detection, respectively. Compared to naïve values, MS-DOHM predicted greater pathogen loads on colonies; however, we found no difference between model estimates and naïve values of prevalence. While the model predicted no declines in site-level prevalence, there were instances where pathogen load decreased in colonies that had been Pd positive for longer periods of time. Our findings demonstrate that accounting for pathogen load and prevalence at multiple scales changes our understanding of Pd dynamics, potentially allowing earlier conservation intervention. Additionally, we found that accounting for pathogen load and prevalence within hibernacula and among individuals resulted in a better fitting model with greater predictive ability.

Scientific Reports

Occupancy modeling for improved accuracy and understanding of pathogen prevalence and dynamics

Most pathogen detection tests are imperfect, with a sensitivity < 100%, thereby resulting in the potential for a false negative, where a pathogen is present but not detected. False negatives in a sample inflate the number of non-detections, negatively biasing estimates of pathogen prevalence. Histological examination of tissues as a diagnostic test can be advantageous as multiple pathogens can be examined and providing important information on associated pathological changes to the host. However, it is usually less sensitive than molecular or microbiological tests for specific pathogens. Our study objectives were to 1) develop a hierarchical occupancy model to examine pathogen prevalence in spring Chinook salmon Oncorhynchus tshawytscha and their distribution among host tissues 2) use the model to estimate pathogen-specific test sensitivities and infection rates, and 3) illustrate the effect of using replicate within host sampling on sample sizes required to detect a pathogen. We examined histological sections of replicate tissue samples from spring Chinook salmon O. tshawytscha collected after spawning for common pathogens seen in this population: Apophallus/ echinostome metacercariae, Parvicapsula minibicornis, Nanophyetus salmincola/ metacercariae, and Renibacterium salmoninarum . A hierarchical occupancy model was developed to estimate pathogen and tissue-specific test sensitivities and unbiased estimation of host- and organ-level infection rates. Model estimated sensitivities and host- and organ-level infections rates varied among pathogens and model estimated infection rate was higher than prevalence unadjusted for test sensitivity, confirming that prevalence unadjusted for test sensitivity was negatively biased. The modeling approach provided an analytical approach for using hierarchically structured pathogen detection data from lower sensitivity diagnostic tests, such as histology, to obtain unbiased pathogen prevalence estimates with associated uncertainties. Accounting for test sensitivity using within host replicate samples also required fewer individual fish to be sampled. This approach is useful for evaluating pathogen or microbe community dynamics when test sensitivity is <100%.

PLoS ONE

Eco‐evolutionary rescue promotes host–pathogen coexistence

Emerging infectious pathogens are responsible for some of the most severe host mass mortality events in wild populations. Yet, effective pathogen control strategies are notoriously difficult to identify, in part because quantifying and forecasting pathogen spread and disease dynamics is challenging. Following an outbreak, hosts must cope with the presence of the pathogen, leading to host–pathogen coexistence or extirpation. Despite decades of research, little is known about host–pathogen coexistence post‐outbreak when low host abundances and cryptic species make these interactions difficult to study. Using a novel disease‐structured N‐mixture model, we evaluate empirical support for three host–pathogen coexistence hypotheses (source–sink, eco‐evolutionary rescue, and spatial variation in pathogen transmission) in a Neotropical amphibian community decimated by Batrachochytrium dendrobatidis ( Bd ) in 2004. During 2010–2014, we surveyed amphibians in Parque Nacional G. D. Omar Torríjos Herrera, Coclé Province, El Copé, Panama. We found that the primary driver of host–pathogen coexistence was eco‐evolutionary rescue, as evidenced by similar amphibian survival and recruitment rates between infected and uninfected hosts. Average apparent monthly survival rates of uninfected and infected hosts were both close to 96%, and the expected number of uninfected and infected hosts recruited (via immigration/reproduction) was less than one host per disease state per 20‐m site. The secondary driver of host–pathogen coexistence was spatial variation in pathogen transmission as we found that transmission was highest in areas of low abundance but there was no support for the source–sink hypothesis. Our results indicate that changes in the host community (i.e., through genetic or species composition) can reduce the impacts of emerging infectious disease post‐outbreak. Our disease‐structured N‐mixture model represents a valuable advancement for conservation managers trying to understand underlying host–pathogen interactions and provides new opportunities to study disease dynamics in remnant host populations decimated by virulent pathogens.

Ecological Applications

Transport and fate of microbial pathogens in agricultural settings

An understanding of the transport and survival of microbial pathogens (pathogens hereafter) in agricultural settings is needed to assess the risk of pathogen contamination to water and food resources, and to develop control strategies and treatment options. However, many knowledge gaps still remain in predicting the fate and transport of pathogens in runoff water, and then through the shallow vadose zone and groundwater. A number of transport pathways, processes, factors, and mathematical models often are needed to describe pathogen fate in agricultural settings. The level of complexity is dramatically enhanced by soil heterogeneity, as well as by temporal variability in temperature, water inputs, and pathogen sources. There is substantial variability in pathogen migration pathways, leading to changes in the dominant processes that control pathogen transport over different spatial and temporal scales. For example, intense rainfall events can generate runoff and preferential flow that can rapidly transport pathogens. Pathogens that survive for extended periods of time have a greatly enhanced probability of remaining viable when subjected to such rapid-transport events. Conversely, in dry seasons, pathogen transport depends more strongly on retention at diverse environmental surfaces controlled by a multitude of coupled physical, chemical, and microbiological factors. These interactions are incompletely characterized, leading to a lack of consensus on the proper mathematical framework to model pathogen transport even at the column scale. In addition, little is known about how to quantify transport and survival parameters at the scale of agricultural fields or watersheds. This review summarizes current conceptual and quantitative models for pathogen transport and fate in agricultural settings over a wide range of spatial and temporal scales. The authors also discuss the benefits that can be realized by improved modeling, and potential treatments to mitigate the risk of waterborne disease transmission.

Critical Reviews in Environmental Science and Tech

Intraspecific and biogeographical variation in foliar fungal communities and pathogen damage of native and invasive Phragmites australis

Aim Recent research has highlighted that the relationship between species interactions and latitude can differ between native and invasive plant taxa, generating biogeographical heterogeneity in community resistance to plant invasions. In the first study with foliar pathogens, we tested whether co‐occurring native and invasive lineages of common reed ( Phragmites australis ) exhibit non‐parallel latitudinal gradients in foliar fungal communities, pathogen susceptibility and damage, and whether these biogeographical patterns can influence the success of invasion. Location North America. Time period 2015–2017. Major taxa studied Perennial grass P. australis . Methods We surveyed 35 P. australis field populations, spanning 17° latitude and comprising four phylogeographical lineages, including one endemic to North America and one invasive from Europe. For each population, we quantified the percentage of leaf pathogen damage and cultured fungi from diseased leaves, which we identified using molecular tools. To assess whether latitudinal gradients in pathogen damage had a genetic basis, we inoculated plants from 73 populations with four putative pathogens in a complementary common garden experiment and measured P. australis susceptibility (i.e., diseased leaf area). Results We isolated 84 foliar fungal taxa. Phragmites australis lineage influenced fungal community composition but not diversity. Despite the invasive European P. australis lineage being the least susceptible to three of the four pathogens tested in the common garden experiment, pathogen damage in the field was similar between native and invasive lineages, providing no evidence that release from foliar pathogens contributes to the success of invasion. Genetically based latitudinal gradients in pathogen susceptibility observed in the common garden were isolate specific and obscured by local environmental conditions in the field, where pathogen damage was threefold higher for northern compared with southern populations, regardless of lineage. Main conclusions Our results highlight that host plant lineage and genetically based biogeographical gradients strongly influence foliar fungal communities and pathogen susceptibility, but do not translate to patterns of pathogen damage observed in the field.

Global Ecology and Biogeography

Future research needs involving pathogens in groundwater

Contamination of groundwater by enteric pathogens has commonly been associated with disease outbreaks. Proper management and treatment of pathogen sources are important prerequisites for preventing groundwater contamination. However, non-point sources of pathogen contamination are frequently difficult to identify, and existing approaches for pathogen detection are costly and only provide semi-quantitative information. Microbial indicators that are readily quantified often do not correlate with the presence of pathogens. Pathogens of emerging concern and increasing detections of antibiotic resistance among bacterial pathogens in groundwater are topics of growing concern. Adequate removal of pathogens during soil passage is therefore critical for safe groundwater extraction. Processes that enhance pathogen transport (e.g., high velocity zones and preferential flow) and diminish pathogen removal (e.g., reversible retention and enhanced survival) are of special concern because they increase the risk of groundwater contamination, but are still incompletely understood. Improved theory and modeling tools are needed to analyze experimental data, test hypotheses, understand coupled processes and controlling mechanisms, predict spatial and/or temporal variability in model parameters and uncertainty in pathogen concentrations, assess risk, and develop mitigation and best management approaches to protect groundwater.

Hydrogeology Journal

Factors related to occurrence and distribution of selected bacterial and protozoan pathogens in Pennsylvania streams

The occurrence and distribution of fecal indicator bacteria (FIB) and bacterial and protozoan pathogens are controlled by diverse factors. To investigate these factors in Pennsylvania streams, 217 samples were collected quarterly from a 27-station water-quality monitoring network from July 2007 through August 2009. Samples were analyzed for concentrations of Escherichia coli (EC) and enterococci (ENT) indicator bacteria, concentrations of Cryptosporidium oocysts and Giardia cysts, and the presence of four genes related to pathogenic types of EC ( eae A, stx 2, stx 1, rfb O157 ) plus three microbial source tracking (MST) gene markers that are also associated with pathogenic ENT and EC ( esp , LTIIa, STII). Water samples were concurrently analyzed for basic water chemistry, physical measures of water quality, nutrients, metals, and a suite of 79 organic compounds that included hormones, pharmaceuticals, and antibiotics. For each sample location, stream discharge was measured by using standardized methods at the time of sample collection, and ancillary sample site information, such as land use and geological characteristics, was compiled. Samples exceeding recreational water quality criteria were more likely to contain all measured pathogen genes but not Cryptosporidium or Giardia (oo)cysts. FIB and Giardia density and frequency of eae A gene occurrence were significantly related to season. When discharge at a sampling location was high (>75th percentile of daily mean discharge), there were greater densities of FIB and Giardia , and the stx2 , rfb O157 , STII, and esp genes were found more frequently than at other discharge conditions. Giardia occurrence was likely related to nonpoint sources, which are highly influential during seasonal overland transport resulting from snowmelt and elevated precipitation in late winter and spring in Pennsylvania. When MST markers of human, swine, or bovine origin were present, samples more frequently carried the eae A, stx 2, stx 1, and rfb O157 genes, but no genes were related exclusively to an individual MST marker. The human source pharmaceuticals (HSPs) acetaminophen and caffeine were correlated with Giardia , and the presence of HSPs proved to be more useful than MST markers in distinguishing the occurrence of Giardia . The HSPs caffeine and carbamazepine were correlated with the sum total of pathogen genes detected in a sample, demonstrating the value of using HSPs as an indicator of fecally derived pathogens. Sites influenced by urban land use with less forest were more likely to have greater FIB and Giardia densities and sum of the array of pathogen genes. Sites dominated by shallow carbonate bedrock in the upstream catchment were likely to have greater FIB densities and higher sum totals of pathogen genes but no correlation with Giardia detection. Our study provides a range of specific environmental, chemical, geologic, and land-use variables related to occurrence and distribution of FIB and selected bacterial and protozoan pathogens in Pennsylvania streams. The information presented could be useful for resource managers in understanding bacterial and protozoan pathogen occurrence and their relation to fecal indicator bacteria in similar settings.

Pennsylvania

Inferring pathogen presence when sample misclassification and partial observation occur

Surveillance programmes are essential for detecting emerging pathogens and often rely on molecular methods to make inference about the presence of a target disease agent. However, molecular methods rarely detect target DNA perfectly. For example, molecular pathogen detection methods can result in misclassification (i.e. false positives and false negatives) or partial detection errors (i.e. detections with ‘ambiguous’, ‘uncertain’ or ‘equivocal’ results). Then, when data are to be analysed, these partial observations are either discarded or censored; this, however, disregards information that could be used to make inference about the true state of the system. There is a critical need for more direction and guidance related to how many samples are enough to declare a unit of interest ‘pathogen free’. Here, we develop a Bayesian hierarchal framework that accommodates false negative, false positive and uncertain detections to improve inference related to the occupancy of a pathogen. We apply our modelling framework to a case study of the fungal pathogen Pseudogymnoascus destructans (Pd) identified in Texas bats at the invasion front of white-nose syndrome. To improve future surveillance programmes, we provide guidance on sample sizes required to be 95% certain a target organism is absent from a site. We found that the presence of uncertain detections increased the variability of resulting posterior probability distributions of pathogen occurrence, and that our estimates of required sample size were very sensitive to prior information about pathogen occupancy, pathogen prevalence and diagnostic test specificity. In the Pd case study, we found that the posterior probability of occupancy was very low in 2018, but occupancy probability approached 1 in 2020, reflecting increasing prior probabilities of occupancy and prevalence elicited from the site manager. Our modelling framework provides the user a posterior probability distribution of pathogen occurrence, which allows for subjective interpretation by the decision-maker. To help readers apply and use the methods we developed, we provide an interactive RShiny app that generates target species occupancy estimation and sample size estimates to make these methods more accessible to the scientific community ( https://rmummah.shinyapps.io/ambigDetect_sampleSize ). This modelling framework and sample size guide may be useful for improving inferences from molecular surveillance data about emerging pathogens, non-native invasive species and endangered species where misclassifications and ambiguous detections occur.

Methods in Ecology and Evolution

Pathogen exposure varies widely among sympatric populations of wild and domestic felids across the United States

Understanding how landscape, host, and pathogen traits contribute to disease exposure requires systematic evaluations of pathogens within and among host species and geographic regions. The relative importance of these attributes is critical for management of wildlife and mitigating domestic animal and human disease, particularly given rapid ecological changes, such as urbanization. We screened >1,000 samples from sympatric populations of puma (Puma concolor), bobcat (Lynx rufus) and domestic cat (Felis catus) across urban gradients in six sites, representing three regions, in North America for exposure to a representative suite of bacterial, protozoal and viral pathogens (Bartonella sp., Toxoplasma gondii, feline herpesvirus-1, feline panleukopenea virus, feline calicivirus, feline immunodeficiency virus). We evaluated prevalence within each species, and examined host trait and land cover determinants of exposure-providing an unprecedented analysis of factors relating to potential for infections in domesticated and wild felids. Prevalence differed among host species (highest for puma and lowest for domestic cat) and was greater for indirectly transmitted pathogens. Sex was inconsistently predictive of exposure to directly transmitted pathogens only, and age infrequently predictive of both direct and indirectly transmitted pathogens. Determinants of pathogen exposure were widely divergent between the wild felid species. For puma, suburban landuse predicted increased exposure to Bartonella sp. in southern California, and FHV-1 exposure increased near urban edges in Florida. This may suggest inter-specific transmission with domestic cats via flea vectors (California) and direct contact (Florida) around urban boundaries. Bobcats captured near urban areas had increased exposure to T. gondii in Florida, suggesting an urban source of prey. Bobcats captured near urban areas in Colorado and Florida had higher FIV exposure, possibly suggesting increased intra-specific interactions through pile-up of home ranges. Beyond these regional and pathogen specific relationships, proximity to the wildland urban interface did not generally increase the probability of disease exposure in wild or domestic felids, emphasizing the importance of local ecological determinants. Indeed, pathogen exposure was often negatively associated with the wildland urban interface for all felids. Our analyses suggest cross-species pathogen transmission events around this interface may be infrequent, but followed by self-sustaining propagation within the new host species.

California, Colorado,Florida