USGS Science⌕ Search

SEARCH · USGS Science

Results for “Pathogens”

Search indexed USGS publications on groundwater, aquifers, geologic maps, mineral resources and earthquakes. Explore source records by subject and place.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 451 records · Page 25Linked to original sources

Variation in gene expression across infection status and elevation in a Hawaiian honeycreeper

Introduced pathogens exert novel selection on hosts, and although many host species have experienced drastic population declines in the absence of adaptation, some hosts have adapted to highly virulent pathogens. For instance, mosquitoes and Plasmodium relictum introduced to the Hawaiian Islands have resulted in extinctions and catastrophic population declines due to avian malaria, particularly in the diverse clade of Hawaiian honeycreepers. However, some species, such as the Hawai'i 'amakihi ( Chlorodrepanis virens ), can survive infection. Immunity exists in low-elevation populations where mosquitoes are abundant, whereas high-elevation, unexposed populations of 'amakihi display greatly reduced immunity. To explore the basis of adaptation to P. relictum in low-elevation 'amakihi, we sequenced transcriptomes from 24 low-elevation and 15 high-elevation 'amakihi. We tested for differential gene expression between (i) infected and uninfected birds and (ii) low- and high-elevation birds. Infected birds showed significant differences in expression across many transcripts with diverse cellular functions involved in different pathways of immune response; eight of the top 13 transcripts blasted to genes previously implicated in immunity to malaria in 'amakihi, and 11 have been identified in other infectious disease systems. Thirteen transcripts showed a trend of higher expression in high-elevation birds. These transcripts blasted to genes involved in metabolism, blood coagulation, and immune response. Our results provide increasing support for a subset of genes involved in immunity to malaria in 'amakihi and hint at possible antagonistic interactions between response to pathogens and environmental characteristics associated with elevation. Further work clarifying the nature of these interactions could benefit conservation efforts of Hawaiian honeycreepers in upper elevation refugia that are increasingly subject to malaria exposure.

Hawaii↗

Spatiotemporal overlap of mallards with poultry farms is associated with greater risk of avian influenza wild bird spillover events

Animal movement influences local transmission and geographic spread of pathogens. Waterfowl are known reservoirs of pathogens, including H5 goose/Guangdong lineage (H5 GsGd) highly pathogenic avian influenza (HPAI). This HPAI virus lineage causes high rates of morbidity and mortality in domestic poultry and many wild bird species. Mallards ( Anas platyrhynchos ) are a generalist waterfowl species whose habitat largely overlaps with many other waterfowl and are considered effective spillover vectors of HPAI. To investigate the potential contribution of waterfowl to HPAI spillover, we used mallards as a proxy and measured the spatiotemporal overlap of 183 GPS-tagged mallards during 2021–2022 with respect to confirmed wild bird spillover events in United States (U.S.) poultry farms. Additionally, we estimated the probability of HPAI spillover events as a function of mallard overlap and poultry farm type. We found infrequent overlap instances between mallards and poultry farms; however, several of these overlap instances lasted > 5 days and up to 19 days. Population-level overlap with poultry farms was greatest during pre-breeding migration, followed by the breeding season. The probability of HPAI spillover was predicted to be greatest for commercial turkey farms, followed by backyard poultry farms. Importantly, farms overlapped by mallards were more than twice as likely to experience a spillover (i.e., increased risk probability), even in the absence of known mallard infection status at the time of overlap. These findings suggest that mallards (and/or other waterfowl) may be important contributors to HPAI spillover into poultry farms and that additional biosecurity measures may be needed. Because few instances of overlap occurred between mallards and farms with reported spillover events, tagged mallards are likely a proxy for other untagged waterfowl. Further studies of wild waterfowl interactions with poultry farms could improve understanding of how landscape characteristics influence spatial overlap, potentially informing which premises may require enhanced biosecurity measures.

Tennessee↗

Dryland fungi are spatially heterogeneous and resistant to global change drivers

Fungi are considered particularly important in regulating the structure and function of dryland ecosystems, yet the response of dryland fungal communities to global change remains notably understudied. Without a clear understanding of how fungi respond to global change drivers, mitigation plans—required for biodiversity and ecosystem service conservation and restoration—are impossible to develop. In this study, we asked the following: (1) How does the fungal community respond to the individual and interactive effects of physical disturbance and drought in a heterogeneous dryland landscape comprised of drought-adapted shrubs separated by adjacent open areas of soil? (2) What are the larger scale impacts of this response? We assessed fungal communities (using fungal-specific DNA metabarcoding analyses) of surface soil samples in an in situ global change experiment that included disturbance and drought in a full factorial design in the northern extent of the Chihuahuan Desert. We found that the fungal community was spatially heterogenous and remarkably resistant to disturbance and drought. We also show that dryland soils harbor high shares of facultative pathogenic and obligately pathogenic fungal taxa, with several concerning taxa reaching high relative abundances under drought. Our results suggest that the fungal community is highly influenced by microclimatic conditions associated with the presence or absence of vegetation. Moreover, our results imply that the fungal community in our experiment was already adapted to the magnitude of stress imposed by two years of experimental disturbance and drought treatments. Overall, our study shows that the fungal community is spatially heterogeneous and resistant to global change drivers and houses many fungal species known for being stress tolerant and pathogenic.

New Mexico↗

Similar population dynamics before and after a chytridiomycosis outbreak in a tropical riparian amphibian species

Emerging infectious diseases can cause rapid, widespread host mortality, and the lack of demographic data before and after pathogen emergence complicates understanding mechanisms of host persistence. This challenge is further compounded by environmental conditions that influence host behavior, while driving pathogen growth and virulence. These interactions create complex disease outcomes that hinder predictions of when and how hosts endure pathogen outbreaks. Here, we analyzed 10 years of capture-mark-recapture data (2000–2014) spanning wet and dry seasons for male Espadarana prosoblepon in El Copé, Panama, encompassing a period before (2000–2004) and after (2010–2014) a Batrachochytrium dendrobatidis ( Bd ) outbreak using Jolly-Seber models. We found that post- Bd male E. prosoblepon population size (range in mean population size among primary periods = 136–225 individuals) was similar to pre- Bd population size (range in mean population size among primary periods = 201–242 individuals). Pre- Bd , average monthly survival probability in the wet season was 0.93 (95% credible interval [CI] = 0.90–0.96). Post- Bd , uninfected individuals had survival probability higher in the wet season (mean = 0.97; [95% CI = 0.95–0.98]) than the dry season (mean = 0.90 [95% CI = 0.84–0.94]), while survival probability for infected individuals decreased as a function of Bd infection intensity. Pre- Bd , mean monthly per-capita entry probability was 0.07 (95% CI = 0.05–0.10), and post- Bd , mean monthly per-capita entry probability was 0.06 (95% CI = 0.00–0.10). Lastly, infection probability during the wet season was lower (mean = 0.04 [95% CI = 0.03–0.05]) than the dry season (mean = 0.10 [95% CI = 0.05–0.15]), and recovery probability during the wet season was lower (mean = 0.19 [95% CI = 0.11–0.28]) than the dry season (mean = 0.54 [95% CI = 0.20–0.88]). Our findings suggest that survival probabilities of uninfected individuals, as well as per-capita entry probabilities, are similar pre- and post- Bd , leading to a stable and similar sized pre- Bd population. These results contribute to understanding disease dynamics and tropical amphibian ecology.

Ecosphere↗

Land use and soil characteristics are associated with increased risk of treponeme-associated hoof disease in elk

Environments can shape the occurrence and extent of disease outbreaks in wildlife. We studied the effects of environmental features on the occurrence of treponeme-associated hoof disease (TAHD), an emerging infectious disease of free-ranging elk ( Cervus canadensis ), in southwestern Washington, USA. During the 2016–2022 harvest seasons, successful elk hunters returned mandatory harvest reports and noted the presence or absence of hoof abnormalities indicative of TAHD. We used generalized linear models and an information-theoretic approach to model selection to relate (1) the spatial distribution of hoof abnormalities to features of landscapes (land cover, topography, and soil characteristics) and (2) the temporal distribution of hoof abnormalities to precipitation during the year preceding the harvest season. The probability of hoof disease increased with soil clay content and proportion of agricultural land (88% of model weight). We found no conclusive evidence for an effect of precipitation on the occurrence of TAHD, but this could relate to relatively high annual precipitation (>140 cm) in the study area. Nevertheless, disease cases may have been negatively associated with precipitation during February–June (55% of model weight). Soils and land management practices may increase the risk of hoof disease by promoting the survival of pathogens that cause TAHD, the susceptibility of elk to infection, or the intensity of pathogen transmission among elk when congregated. Focusing on areas where the risk of disease is greatest may facilitate the detection of TAHD during surveillance. Likewise, removing infected elk and dispersing uninfected elk from areas with the greatest risk of disease may enhance the effectiveness of efforts to reduce transmission. Basing this work on the knowledge that disease risk is modified by factors of hosts, pathogens, and environments, this study serves as an application of the epidemiological triad framework to better understand the ecology and epidemiology of an emerging infectious disease in wildlife.

Washington↗

Ecology of reintroduced Rocky Mountain bighorn sheep in Dinosaur National Monument

Translocations have been widely used to restore and conserve bighorn sheep ( Ovis canadensis ) populations in North America. Some translocations have been successful, but many populations remain small and genetically isolated. Population structure can influence the viability and long-term success of reintroductions. Social ungulates often function as interconnected subpopulations (metapopulations); however, few studies evaluate subpopulation sizes, connectivity, and genetic diversity within metapopulations. To address this gap, we conducted a comprehensive study of a reintroduced Rocky Mountain bighorn sheep ( Ovis canadensis canadensis ) population in Dinosaur National Monument in Colorado and Utah, USA, between 2006–2020. We analyzed global positioning system (GPS) radio-collar data, genetic samples, and results of health testing to evaluate abundance, distribution, genetic structure and diversity, habitat use, movement and connectivity, and presence of or exposure to respiratory pathogens. We integrated these analyses to evaluate the outcomes of a reintroduction effort that began in 1952, over 70 years ago, and to inform management decisions in Dinosaur National Monument. We also provide a framework for evaluating metapopulation processes, including a non-invasive approach that links genetic structure with Bayesian spatial capture-recapture analyses to estimate subpopulation sizes. Despite models indicating continuous suitable habitat, we found a spatially structured population with at least 4 subpopulations with constrained connectivity. Evidence from step selection and density analyses suggested that movement among subpopulations may be limited by semi-permeable barriers including rivers and human disturbance, which could contribute to maintenance of spatial structure over time. In 2006, antibody to Mycoplasma ovipneumoniae was detected in all geographically and genetically distinct subpopulations. Widespread clinical signs of disease and a confirmed exposure to M. ovipneumoniae in 2019 indicate a long-term disease challenge. Proximity to domestic sheep creates repeated opportunities for introduction of new M. ovipneumoniae strains. We estimated abundance in 2019 at 109 (95% CrI = 87–133), composed of subpopulations ranging from 18–39 animals (95% CrIs from 11–50). Genetic diversity was relatively high compared to other reintroduced and native Rocky Mountain bighorn sheep populations, which is likely a consequence of multiple translocations from different sources. Three of 4 subpopulation centers generally aligned with the locations of original translocation release sites. Persistence in the presence of pathogens may be facilitated by metapopulation structure and moderately high genetic diversity. Conversely, metapopulation structure can also facilitate pathogen persistence. Our approach offers a path to advance understanding of the population ecology of reintroduced bighorn sheep and can inform effective conservation and management of their populations.

Colorado, Wyoming↗

Batrachochytrium salamandrivorans and the risk of a second amphibian pandemic

Amphibians are experiencing devastating population declines globally. A major driver is chytridiomycosis, an emerging infectious disease caused by the fungal pathogens Batrachochytrium dendrobatidis ( Bd ) and Batrachochytrium salamandrivorans ( Bsal ). Bd was described in 1999 and has been linked with declines since the 1970s, while Bsal is a more recently discovered pathogen that was described in 2013. It is hypothesized that Bsal originated in Asia and spread via international trade to Europe, where it has been linked to salamander die-offs. Trade in live amphibians thus represents a significant threat to global biodiversity in amphibians. We review the current state of knowledge regarding Bsal and describe the risk of Bsal spread. We discuss regional responses to Bsal and barriers that impede a rapid, coordinated global effort. The discovery of a second deadly emerging chytrid fungal pathogen in amphibians poses an opportunity for scientists, conservationists, and governments to improve global biosecurity and further protect humans and wildlife from a growing number of emerging infectious diseases.

EcoHealth↗

Functional role of an endophytic Bacillus amyloliquefaciens in enhancing growth and disease protection of invasive English ivy ( Hedera helix L. )

Background We hypothesize that invasive English ivy ( Hedera helix ) harbors endophytic microbes that promote plant growth and survival. To evaluate this hypothesis, we examined endophytic bacteria in English ivy and evaluated effects on the host plant. Methods Endophytic bacteria were isolated from multiple populations of English ivy in New Brunswick, NJ. Bacteria were identified as a single species Bacillus amyloliquefaciens . One strain of B. amyloliquefaciens , strain C6c, was characterized for indoleacetic acid (IAA) production, secretion of hydrolytic enzymes, phosphate solubilization, and antibiosis against pathogens. PCR was used to amplify lipopeptide genes and their secretion into culture media was detected by MALDI-TOF mass spectrometry. Capability to promote growth of English ivy was evaluated in greenhouse experiments. The capacity of C6c to protect plants from disease was evaluated by exposing B+ (bacterium inoculated) and B− (non-inoculated) plants to the necrotrophic pathogen Alternaria tenuissima . Results B. amyloliquefaciens C6c systemically colonized leaves, petioles, and seeds of English ivy. C6c synthesized IAA and inhibited plant pathogens. MALDI-TOF mass spectrometry analysis revealed secretion of antifungal lipopeptides surfactin, iturin, bacillomycin, and fengycin. C6c promoted the growth of English ivy in low and high soil nitrogen conditions. This endophytic bacterium efficiently controlled disease caused by Alternaria tenuissima . Conclusions This study suggests that B. amyloliquefaciens plays an important role in enhancing growth and disease protection of English ivy.

Plant and Soil↗

Chapter 5: Health and diseases

Health and diseases are integral parts of the life of seabirds that merit attention if we expect to truly understand, protect, and conserve them. Diseases such as avian influenza, avian pox, pasteurellosis, and paralytic shellfish poisoning have a proven history of decreasing the survival or breeding success of seabirds. However, each host-pathogen-environment system is unique, and our current knowledge about seabird health is limited and subject to biases. Thus, an exploratory mindset should be maintained, always considering that new or previously undiagnosed diseases could have substantial effects on a given seabird population. Therefore, incorporating a health monitoring component in seabird population monitoring programs, wherein data and biological samples are routinely collected for long-term pathogen surveillance and physiological analyses, would help us understand factors that limit seabird populations. Finally, the implementation of biosecurity best practices at seabird aggregations is imperative to avoid the accidental introduction or spread of pathogens.

Book chapter↗

Livestock grazing, climatic variation, and breeding phenology jointly shape disease dynamics and survival in a wild amphibian

Wildlife responses to infectious disease can be influenced by environmental stressors that alter host-pathogen dynamics. We investigated how livestock grazing, climatic variation, and breeding phenology influence disease prevalence and annual survival in boreal toad ( Anaxyrus boreas boreas ) populations challenged with Batrachochytrium dendrobatidis ( Bd ), a fungal pathogen implicated in global amphibian declines. We conducted a five-year (2015–2019) capture-recapture study of boreal toads ( n = 1301) inhabiting pastures grazed by cattle in western Wyoming, USA. We employed structural equation models to determine whether the effects of climatic variation on Bd prevalence were direct or mediated through effects on breeding phenology and multi-state models to explore the interplay of grazing, weather, and Bd infection on adult survival. Higher winter snowpack was linked with shorter spring breeding seasons, which were associated with lower Bd prevalence. Boreal toads infected with Bd suffered increased mortality, but only at relatively cool temperatures. Although cattle grazing created warmer microclimates, likely by reducing vegetation cover, grazing-induced habitat changes did not scale up to influence adult survival. Our results suggest that boreal toads in cooler environments face increased risk of disease-induced mortality, possibly because infected individuals are not able to elevate body temperature to reduce or clear infection. More generally, we demonstrate that host-pathogen dynamics can be shaped jointly by independent and interactive effects of livestock grazing, breeding season length, and climatic variation. Future investigations of wildlife responses to disease therefore may benefit from considering anthropogenic land use and climatic regimes, including the effect of weather on host phenology.

Wyoming↗

An epidemiological model of virus transmission in salmonid fishes of the Columbia River Basin

We have developed a dynamic epidemiological model informed by records of viral presence and genotypes to evaluate potential transmission routes maintaining a viral pathogen in economically and culturally important anadromous fish populations. In the Columbia River Basin, infectious hematopoietic necrosis virus (IHNV) causes severe disease, predominantly in juvenile steelhead trout ( Oncorhynchus mykiss ) and less frequently in Chinook salmon ( O. tshawytscha ). Mortality events following IHNV infection can be devastating for individual hatchery programs. Despite reports of high local mortality and extensive surveillance efforts, there are questions about how viral transmission is maintained. Modeling this system offers important insights into disease transmission in natural aquatic systems, as well as about the data requirements for generating accurate estimates about transmission routes and infection probabilities. We simulated six scenarios in which testing rates and the relative importance of different transmission routes varied. The simulations demonstrated that the model accurately identified routes of transmission and inferred infection probabilities accurately when there was testing of all cohort-sites. When testing records were incomplete, the model accurately inferred which transmission routes exposed particular cohort-sites but generated biased infection probabilities given exposure. After validating the model and generating guidelines for result interpretation, we applied the model to data from 14 annual cohorts (2000–2013) at 24 focal sites in a sub-region of the Columbia River Basin, the lower Columbia River (LCR), to quantify the relative importance of potential transmission routes in this focal sub-region. We demonstrate that exposure to IHNV via the return migration of adult fish is an important route for maintaining IHNV in the LCR sub-region, and the probability of infection following this exposure was relatively high at 0.16. Although only 1% of cohort-sites experienced self-exposure by infected juvenile fish, this transmission route had the greatest probability of infection (0.22). Increased testing and/or determining whether transmission can occur from cohort-sites without testing records (e.g., determining there was no testing record because there were no fish at the cohort-site) are expected to improve inference about infection probabilities. Increased use of secure water supplies and continued use of biosecurity protocols may reduce IHNV transmission from adult fish and juvenile fish within the site, respectively, to juvenile salmonids at hatcheries. Models and conclusions from this study are potentially relevant to understanding the relative importance of transmission routes for other important aquatic pathogens in salmonids, including the agents of bacterial kidney disease and coldwater disease, and the basic approach may be useful for other pathogens and hosts in other geographic regions.

Ecological Modelling↗

Biomedical and veterinary science can increase our understanding of coral disease

A balanced approach to coral disease investigation is critical for understanding the global decline of corals. Such an approach should involve the proper use of biomedical concepts, tools, and terminology to address confusion and promote clarity in the coral disease literature. Investigating disease in corals should follow a logical series of steps including identification of disease, systematic morphologic descriptions of lesions at the gross and cellular levels, measurement of health indices, and experiments to understand disease pathogenesis and the complex interactions between host, pathogen, and the environment. This model for disease investigation is widely accepted in the medical, veterinary and invertebrate pathology disciplines. We present standard biomedical rationale behind the detection, description, and naming of diseases and offer examples of the application of Koch's postulates to elucidate the etiology of some infectious diseases. Basic epidemiologic concepts are introduced to help investigators think systematically about the cause(s) of complex diseases. A major goal of disease investigation in corals and other organisms is to gather data that will enable the establishment of standardized case definitions to distinguish among diseases. Concepts and facts amassed from empirical studies over the centuries by medical and veterinary pathologists have standardized disease investigation and are invaluable to coral researchers because of the robust comparisons they enable; examples of these are given throughout this paper. Arguments over whether coral diseases are caused by primary versus opportunistic pathogens reflect the lack of data available to prove or refute such hypotheses and emphasize the need for coral disease investigations that focus on: characterizing the normal microbiota and physiology of the healthy host; defining ecological interactions within the microbial community associated with the host; and investigating host immunity, host-agent interactions, pathology, pathogenesis, and factors that promote the pathogenicity of the causative agent(s) of disease.

Journal of Experimental Marine Biology and Ecology↗

Identification and characterization of Highlands J virus from a Mississippi sandhill crane using unbiased next-generation sequencing

Advances in massively parallel DNA sequencing platforms, commonly termed next-generation sequencing (NGS) technologies, have greatly reduced time, labor, and cost associated with DNA sequencing. Thus, NGS has become a routine tool for new viral pathogen discovery and will likely become the standard for routine laboratory diagnostics of infectious diseases in the near future. This study demonstrated the application of NGS for the rapid identification and characterization of a virus isolated from the brain of an endangered Mississippi sandhill crane. This bird was part of a population restoration effort and was found in an emaciated state several days after Hurricane Isaac passed over the refuge in Mississippi in 2012. Post-mortem examination had identified trichostrongyliasis as the possible cause of death, but because a virus with morphology consistent with a togavirus was isolated from the brain of the bird, an arboviral etiology was strongly suspected. Because individual molecular assays for several known arboviruses were negative, unbiased NGS by Illumina MiSeq was used to definitively identify and characterize the causative viral agent. Whole genome sequencing and phylogenetic analysis revealed the viral isolate to be the Highlands J virus, a known avian pathogen. This study demonstrates the use of unbiased NGS for the rapid detection and characterization of an unidentified viral pathogen and the application of this technology to wildlife disease diagnostics and conservation medicine.

Journal of Virological Methods↗

Host mating system and the spread of a disease-resistant allele in a population

The model presented here modifies a susceptible-infected (SI) host-pathogen model to determine the influence of mating system on the outcome of a host-pathogen interaction. Both deterministic and stochastic (individual-based) versions of the model were used. This model considers the potential consequences of varying mating systems on the rate of spread of both the pathogen and resistance alleles within the population. We assumed that a single allele for disease resistance was sufficient to confer complete resistance in an individual, and that both homozygote and heterozygote resistant individuals had the same mean birth and death rates. When disease invaded a population with only an initial small fraction of resistant genes, inbreeding (selfing) tended to increase the probability that the disease would soon be eliminated from a small population rather than become endemic, while outcrossing greatly increased the probability that the population would become extinct due to the disease.

Theoretical Population Biology↗

Molecular detection of bacteria in the families Rickettsiaceae and Anaplasmataceae in northern crested caracaras ( Caracara cheriway )

Bacterial pathogens of the families Anaplasmataceae and Rickettsiaceae are often spread to humans or other animals from bites from infected arthropod hosts. Recently, an increasing number of studies have implicated migratory birds in the circulation of these pathogens through the spread of arthropod vectors. However, few studies have examined the potential for resident bird populations to serve as reservoirs for these zoonoses. In this study, we used nested PCRs of the GroESL and 17 kDa genes to screen for Anaplasmataceae and Rickettsiaceae , respectively, in a resident population of the northern crested caracara ( Caracara cheriway ) from Florida ( n = 55). Additionally, a small number ( n = 6) of captive individuals from Texas were included. We identified one individual (1.64%) positive for Rickettsia felis and one (1.64%) positive for Ehrlichia chaffeensis; both these individuals were from Florida. Presence of these pathogens demonstrates that these birds are potential hosts; however, the low prevalence of infections suggests that these populations likely do not function as an ecological reservoir.

Ticks and Tick-borne Diseases↗

Biofilm formation of Francisella noatunensis subsp. orientalis

Francisella noatunensis subsp. orientalis ( Fno ) is an emergent fish pathogen in both marine and fresh water environments. The bacterium is suspected to persist in the environment even without the presence of a suitable fish host. In the present study, the influence of different abiotic factors such as salinity and temperature were used to study the biofilm formation of different isolates of Fno including intracellular growth loci C (iglC) and pathogenicity determinant protein A (pdpA) knockout strains. Finally, we compared the susceptibility of planktonic and biofilm to three disinfectants used in the aquaculture and ornamental fish industry, namely Virkon ® , bleach and hydrogen peroxide. The data indicates that Fno is capable of producing biofilms within 24 h where both salinity as well as temperature plays a role in the growth and biofilm formation of Fno . Mutations in the iglC or pdpA , both known virulence factors, do not appear to affect the capacity of Fno to produce biofilms, and the minimum inhibitory concentration, and minimum biocidal concentration for the three disinfectants were lower than the minimum biofilm eradication concentration values. This information needs to be taken into account if trying to eradicate the pathogen from aquaculture facilities or aquariums.

Veterinary Microbiology↗

Detection, quantitation and identification of enteroviruses from surface waters and sponge tissue from the Florida Keys using real-time RT-PCR

A method was developed for the quantitative detection of pathogenic human enteroviruses from surface waters in the Florida Keys using Taqman (R) one-step Reverse transcription (RT)-PCR with the Model 7700 ABI Prism (R) Sequence Detection System. Viruses were directly extracted from unconcentrated grab samples of seawater, from seawater concentrated by vortex flow filtration using a 100kD filter and from sponge tissue. Total RNA was extracted from the samples, purified and concentrated using spin-column chromatography. A 192-196 base pair portion of the 5??? untranscribed region was amplified from these extracts. Enterovirus concentrations were estimated using real-time RT-PCR technology. Nine of 15 sample sites or 60% were positive for the presence of pathogenic human enteroviruses. Considering only near-shore sites, 69% were positive with viral concentrations ranging from 9.3viruses/ml to 83viruses/g of sponge tissue (uncorrected for extraction efficiency). Certain amplicons were selected for cloning and sequencing for identification. Three strains of waterborne enteroviruses were identified as Coxsackievirus A9, Coxsackievirus A16, and Poliovirus Sabin type 1. Time and cost efficiency of this one-step real-time RT-PCR methodology makes this an ideal technique to detect, quantitate and identify pathogenic enteroviruses in recreational waters. Copyright ?? 2002 Elsevier Science Ltd.

Water Research↗

Robotic environmental DNA bio-surveillance of freshwater health

Autonomous water sampling technologies may help to overcome the human resource challenges of monitoring biological threats to rivers over long time periods and large geographic areas. The Monterey Bay Aquarium Research Institute has pioneered a robotic Environmental Sample Processor (ESP) that overcomes some of the constraints associated with traditional sampling since it can automate water sample filtration and preservation of the captured material or homogenize it for immediate analyses in situ. The ESP was originally developed for marine environment applications. Here we evaluated whether the ESP can provide reliable, timely information on environmental (e)DNA detections of human and fish pathogens and introduced fishes at U.S. Geological Survey streamgage sites in freshwater rivers. We compared eDNA collected via ESP at high frequency (e.g., every 3 h) with manual eDNA collections collected at lower frequency (e.g., weekly). We found that water samples filtered and preserved by ESPs successfully detected the DNA of human pathogens, fish pathogens and introduced fishes. Both ESP and manually collected samples provided similar information about target DNA presence. We suggest that the greatest current benefit of the ESP is the cost savings of high frequency, bio-surveillance at remote or hard to access sites. The full potential of robotic technologies like the ESP will be realized when they can more easily execute in situ analyses of water samples and rapidly transmit results to decision-makers.

Idaho, Montana, Wyoming↗