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William N. Batts

Publications and source records attributed to William N. Batts.

At least 19 recordsLinked to original sources

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

Virulence evolution of a salmonid virus following a host jump

Emergent viral diseases remain a critical obstacle to welfare across landscapes and species, encompassing humans, wildlife, and agriculture. Following a jump to a novel host, the severity of disease resulting from infection is a critical determinant of the overall emergent pathogen threat. Conventional wisdom posits that virulence, defined here as host mortality, attenuates to intermediate levels as a pathogen adapts to a novel host, but this is largely based on data from just one system, myxoma virus, which was intentionally introduced as a biocontrol agent in rabbits ( Oryctolagus cuniculus ) in mid-1900s Australia. In this study, we demonstrate that infectious hematopoietic necrosis virus (IHNV), which made a host jump from sockeye salmon ( Oncorhynchus nerka , ancestral host) to rainbow trout ( O. mykiss , novel host), has not conformed to classical theory. We quantified virulence in the ancestral and novel hosts using common garden in vivo experiments with 16 archival IHNV isolates collected from 1972-2017, which span the period from shortly after the host jump and the subsequent 45 years of host adaptation. These virus isolates also represent two distinct phylogenetic genogroups, each associated with either the ancestral or novel host. The experiments were replicated across two research facilities, two challenges dosages, and two temperatures. While isolates from the ancestral genogroup showed no temporal change in virulence in either host, isolates from the novel viral genogroup displayed a significant increase in virulence over time in the novel host. Some possible indication of a virus temperature adaption after the host jump was also present. Potential drivers of virulence evolution are discussed. This represents one of only a handful of systems in which the evolution of increased virulence has been empirically characterized after a host jump and subsequent adaptation. It contributes to a growing body of evidence that contradicts the classical case study of myxoma virus attenuation after adaptation.

PLoS Pathogens

Evaluation of 6PPD-quinone lethal toxicity and sublethal effects on disease resistance and swimming fitness in coastal cutthroat trout (Oncorhynchus clarkii clarkii)

6PPD-quinone (6PPDQ), derived from the tire-protectant 6PPD reacting with ozone, is an emerging contaminant of concern owing to its role in coho salmon ( Oncorhynchus kisutch ) deaths via urban runoff mortality syndrome (URMS). Given the impact of 6PPDQ on aquatic life in urban streams, we addressed the acute toxicity of 6PPDQ exposure on coastal cutthroat trout (CCT) ( Oncorhynchus clarkii clarkii ), a species sympatric with coho salmon in natal watersheds. Using static exposures coupled with analytical chemistry, we determined the 24-h LC 50 values for alevin (297.2 ng/L), swim-up fry (39.6 ng/L), 5-month parr (103.3 ng/L), and 13-month juveniles (185.9 ng/L)─values similar to toxicity observed in coho salmon. Additionally, the 96-h LC 50 (77.6 ng/L) was 2.4 times more lethal for juvenile CCT. We assessed potential effects of sublethal 6PPDQ exposure on disease resistance to infectious hematopoietic necrosis (IHN), an endemic viral disease of Pacific salmon, and to swimming performance. Sublethal 6PPDQ (53.6 ng/L) did not affect survival of parr exposed to IHN virus compared to virus alone. Conversely, 6PPDQ exposure as low as 72.2 ng/L significantly reduced 15- and 24-month juvenile swimming performance, and 120.5 ng/L 6PPDQ increased blood hematocrit. Overall, CCT are the second most sensitive species tested to date for 6PPDQ sensitivity which further emphasizes the need for identifying alternatives to 6PPD.

Environmental Science and Technology

Hepeviruses of aquatic organisms

Originally reported in California, the cutthroat trout virus (CTV) has now been isolated from nine species of salmonids in North America. Early work focused on the replication and physical characteristics of the virus, but 20 years later was determined to be most closely related to the hepatitis E virus. The small genome is positive-sense, single-stranded RNA similar to other members of the family Hepeviridae , which now contains its own genus Piscihepevirus with two distinct genotypes, CTV-1 and CTV-2. While CTV has not been associated with acute disease in fish, the virus could form persistently infected cell cultures that may aid research in treating hepatitis E-like viruses affecting humans or other animals. Interestingly, trout exposed to CTV were protected for about a month against subsequent exposure to the infectious hematopoietic necrosis virus. Replicating agents suspected to be CTV can be confirmed by polymerase chain reaction (PCR), quantitative PCR, and sequencing. Other unclassified hepeviruses detected in fish using viral metagenomics include Wenling fish hepevirus, Wenling moray eel hepevirus, Murray–Darling carp hepevirus, and eastern mosquitofish hepevirus. The family Hepeviridae has been placed in the order Hepevirales together with the family Matonaviridae (rubella virus), with member viruses having amino acid homology in the helicase and replicase regions of the nonstructural proteins. In addition, using next-generation sequencing, a hepe-like sequence was characterized in diseased giant freshwater prawn Macrobrachium rosenbergii and named Crustacea hepe-like virus 1. Thus the family Hepeviridae continues to expand among aquatic animal hosts.

Book chapter

Paramyxoviruses of fish

The first fish paramyxovirus was isolated from normal adult Chinook salmon returning to a coastal hatchery in Oregon in the fall of 1982. Subsequently, the virus was isolated from other stocks of adult Chinook salmon and one stock of adult coho salmon in California, Oregon, Washington, and Alaska, leading to its designation as the Pacific salmon paramyxovirus (PsaPV). The slow-growing virus can be isolated from tissues and ovarian fluids of healthy adult fish returning to spawn and apparently causes no clinical signs of disease or mortality. In 1995 a different and widely disseminated paramyxovirus was isolated from farmed Atlantic salmon in Norway and was designated as Atlantic salmon paramyxovirus (AsaPV). Although this virus caused no disease or mortality when injected into juvenile Atlantic salmon, AsaPV has been associated with proliferative gill inflammation in sea-reared yearling fish; however, additional infectious agents may be involved in the etiology of the condition. Sequence analysis of PsaPV and AsaPV isolates using the polymerase gene established their placement in the family Paramyxoviridae and has shown the two viruses to be closely related but sufficiently different from each other and from other known paramyxoviruses to represent a new genus within the family. The viruses can be diagnosed by isolation in cell culture with final confirmation by molecular methods. Other paramyxovirus-like agents have been observed or isolated from rainbow trout in Germany, from seabream in Japan associated with epithelial necrosis, from turbot in Spain associated with erythrocytic inclusion bodies and buccal/opercular hemorrhaging and from koi and common carp associated with gill necrosis in the European Union.

Book chapter

Genetics reveal long-distance virus transmission links in Pacific salmon

In the coastal region of Washington State, a major pathogen emergence event occurred between 2007 and 2011 in which steelhead trout ( Oncorhynchus mykiss ) experienced a high incidence of infection and disease outbreaks due to the rhabdovirus infectious hematopoietic necrosis virus (IHNV). Genetic typing showed that the introduced viruses were in the steelhead-specific MD subgroup of IHNV and indicated the most likely source was a virus from the nearby Columbia River Basin. In the current study, full-length viral glycoprotein (G) gene sequences were determined for 55 IHNV isolates from both coastal and Columbia fish populations to identify specific source populations and infer mechanisms of transmission to coastal steelhead. We identified three transmission links based on exact fullG genotype matches between Columbia and coastal fish. In all cases, the likely source population was infected juvenile fish, and sink populations were adult fish returning to coastal rivers to spawn. The time intervals between detection in source and sink populations varied from 6 months to nearly 4 years, suggesting different transmission pathways. Surprisingly, distances between source and sink populations varied between 140 and 1000 km. These results confirm repeated introductions of virus from Columbia River Basin fish as the cause of emergence of MD virus on the Washington coast from 2007 to 2011. View Full-Text

Washington

Rapid diagnostic test to detect and discriminate infectious hematopoietic necrosis virus (IHNV) genogroups U and M to aid management of Pacific Northwest salmonid populations

Infectious hematopoietic necrosis virus (IHNV) is an acute pathogen of salmonids in North America, Europe, and Asia that is phylogenetically classified into five major virus genogroups (U, M, L, E, and J). The geographic range of the U and M genogroup isolates overlap in the North American Columbia River Basin and Washington Coast region, where these genogroups pose different risks depending on the species of Pacific salmon ( Oncorhynchus spp.). For certain management decisions, there is a need to both test for IHNV presence and rapidly determine the genogroup. Herein, we report the development and validation of a U/M multiplex reverse transcription, real-time PCR (RT-rPCR) assay targeting the IHNV nucleocapsid (N) protein gene. The new U/M RT-rPCR is a rapid, sensitive, and repeatable assay capable of specifically discriminating between North American U and M genogroup IHNV isolates. However, one M genogroup isolate obtained from commercially cultured Idaho rainbow trout ( O. mykiss ) showed reduced sensitivity with the RT-rPCR test, suggesting caution may be warranted before applying RT-rPCR as the sole surveillance test in areas associated with the Idaho trout industry. The new U/M assay had high diagnostic sensitivity (DSe > 94%) and specificity (DSp > 97%) in free-ranging adult Pacific salmon, when assessed relative to cell culture, the widely accepted reference standard, as well as the previously validated universal N RT-rPCR test. The high diagnostic performance of the new U/M assay indicates the test is suitable for surveillance, diagnosis, and confirmation of IHNV in Pacific salmon from the Pacific Northwest regions where the U and M genogroups overlap.

Animals

Novel diagnostic tests for the putative agent of bacterial gill disease in Pacific razor clams (Siliqua patula)

Nuclear inclusion X (NIX) is a gamma proteobacteria that infects the nuclei of gill epithelial cells in Pacific razor clams. NIX has been associated with clam die-offs in coastal Washington. A quantitative PCR (qPCR) assay was developed to detect NIX in Pacific razor clams, and assay specificity was confirmed by chromogenic in situ hybridization (CISH). Both tests were applied to evaluate NIX infections in wild Pacific razor clams collected during spring 2019. Consistent with results from earlier histopathological assessments, qPCR and CISH indicated 100% prevalence in razor clams from two Washington beaches and 0% prevalence from two Alaskan beaches.

Alaska, Washington

Ichthyophonus sp. Infection in Opaleye (Girella nigricans)

Over a 3-year-period, 17 wild-caught opaleye (Girella nigricans) housed in a public display aquarium were found dead without premonitory signs. Grossly, 4 animals had pinpoint brown or black foci on coelomic adipose tissue. Histologically, liver, spleen, heart, and posterior kidney had mesomycetozoan granulomas in all cases; other organs were less commonly infected. Four opaleye had goiter; additional substantial lesions were not identified. Granulomas surrounded melanized debris, leukocytes, and mesomycetozoa represented by folded membranes (collapsed schizont walls), intact schizonts (50- to >200 µm in diameter with a multilaminate membrane), plasmodia (budding from schizonts or free in tissue), or rarely germinal tubes (budding from schizonts). Ichthyophonus was grown from fresh tissues in tissue explant broth cultures of the heart, liver, and/or spleen. Polymerase chain reaction using 18S ribosomal DNA primers amplified a 1730-bp region, and the DNA sequence was most similar to Ichthyophonus hoferi, which is often associated with freshwater aquaculture fish.

Veterinary Pathology

Anadromous coastal cutthroat trout Oncorhynchus clarkii clarkii as a host for Argulus pugettensis (Crustacea, Branchiura): Parasite prevalence, intensity and distribution

Coastal cutthroat trout [ Oncorhynchus clarkii clarkii (Richardson, 1836)] from the marine waters of Puget Sound, WA, was documented as a new host for the ectoparasite Argulus pugettensis ( Dana, 1852 ). The prevalence of A. pugettensis was 66% (49 of 74) on cutthroat trout and 0% (0 of 55) on coho salmon [ O. kisutch (Walbaum, 1792)] collected during the winter of 2017/2018. Infestations occurred most frequently on the dorsal surface, with intensities ranging from 1 to 26 argulids per fish (mean intensity 3.94 ± 4.93 S.D.). In contrast, the prevalence of the common salmon louse [ Lepeophtheirus salmonis (Krøyer, 1837)] was 72% for cutthroat trout and 31% for coho salmon. Relative to other native salmonids, little is known regarding the status, ecology and threats for coastal cutthroat trout. New information reported here is a first step in understanding the relationship between this wild, native trout and infestations by parasitic sea lice and should be followed by future studies aimed to identify population level consequences.

Northwest Science

Development and characterization of two cell lines from gills of Atlantic salmon

Gill disease in Atlantic salmon, Salmo salar L., causes big losses in the salmon farming industry. Until now, tools to cultivate microorganisms causing gill disease and models to study the gill responses have been lacking. Here we describe the establishment and characterization of two cell lines from the gills of Atlantic salmon. Atlantic salmon gill cell ASG-10 consisted of cells staining for cytokeratin and e-cadherin and with desmosomes as seen by transmission electron microscopy suggesting the cells to be of epithelial origin. These structures were not seen in ASG-13. The cell lines have been maintained for almost 30 passages and both cell lines are fully susceptible to infection by infectious hematopoietic necrosis virus (IHNV), viral hemorrhagic septicemia virus (VHSV), infectious pancreatic necrosis virus (IPNV), Atlantic salmon reovirus TS (TSRV) and Pacific salmon paramyxovirus (PSPV). While infectious salmon anemia virus (ISAV) did not cause visible CPE, immunofluorescent staining revealed a sub-fraction of cells in both the ASG-10 and ASG-13 lines may be permissive to infection. ASG-10 is able to proliferate and migrate to close scratches in the monolayer within seven days in vitro contrary to ASG-13, which does not appear to do have the same proliferative and migratory ability. These cell lines will be useful in studies of gill diseases in Atlantic salmon and may represent an important contribution for alternatives to experimental animals and studies of epithelial–mesenchymal cell biology.

PLoS ONE

Isolation and characterization of the fall Chinook aquareovirus

Background Salmon are paramount to the economy, ecology, and history of the Pacific Northwest. Viruses constitute one of the major threats to salmon health and well-being, with more than twenty known virus species that infect salmon. Here, we describe the isolation and characterization of the fall Chinook aquareovirus, a divergent member of the species Aquareovirus B within the family Reoviridae . Methods The virus was first found in 2014 as part of a routine adult broodstock screening program in which kidney and spleen tissue samples from healthy-appearing, adult fall Chinook salmon ( Oncorhynchus tshawytscha ) returning to a hatchery in Washington State produced cytopathic effects when inoculated onto a Chinook salmon embryo cell line (CHSE-214). The virus was not able to be confirmed by an RT-PCR assay using existing aquareovirus pan-species primers, and instead was identified by metagenomic next-generation sequencing. Metagenomic next-generation sequencing was used to recover the full genome and completed using 3′ RACE. Results The genome of the fall Chinook aquareovirus contains 11 segments of double-stranded RNA totaling 23.3 kb, with each segment flanked by the canonical sequence termini found in the aquareoviruses. Sequence comparisons and a phylogenetic analysis revealed a nucleotide identity of 63.2% in the VP7 gene with the Green River Chinook virus, placing the new isolate in the species Aquareovirus B . A qRT-PCR assay was developed targeting the VP2, which showed rapid growth of the isolate during the initial 5 days in culture using CHSE-214 cells. Conclusions This sequence represents the first complete genome of an Aquareovirus B species. Future studies will be required to understand the potential pathogenicity and epidemiology of the fall Chinook aquareovirus.

Virology Journal

Molecular characterization of a novel orthomyxovirus from rainbow and steelhead trout ( Oncorhynchus mykiss )

A novel virus, rainbow trout orthomyxovirus (RbtOV), was isolated in 1997 and again in 2000 from commercially-reared rainbow trout ( Oncorhynchus mykiss ) in Idaho, USA. The virus grew optimally in the CHSE-214 cell line at 15°C producing a diffuse cytopathic effect; however, juvenile rainbow trout exposed to cell culture-grown virus showed no mortality or gross pathology. Electron microscopy of preparations from infected cell cultures revealed the presence of typical orthomyxovirus particles. The complete genome of RbtOV is comprised of eight linear segments of single-stranded, negative-sense RNA having highly conserved 5′ and 3′-terminal nucleotide sequences. Another virus isolated in 2014 from steelhead trout (also O. mykiss ) in Wisconsin, USA, and designated SttOV was found to have eight genome segments with high amino acid sequence identities (89–99%) to the corresponding genes of RbtOV, suggesting these new viruses are isolates of the same virus species and may be more widespread than currently realized. The new isolates had the same genome segment order and the closest pairwise amino acid sequence identities of 16–42% with Infectious salmon anemia virus (ISAV), the type species and currently only member of the genus Isavirus in the family Orthomyxoviridae . However, pairwise comparisons of the predicted amino acid sequences of the 10 RbtOV and SttOV proteins with orthologs from representatives of the established orthomyxoviral genera and a phylogenetic analysis using the PB1 protein showed that while RbtOV and SttOV clustered most closely with ISAV, they diverged sufficiently to merit consideration as representatives of a novel genus. A set of PCR primers was designed using conserved regions of the PB1 gene to produce amplicons that may be sequenced for identification of similar fish orthomyxoviruses in the future.

Virus Research

Complete sequences of 4 viral hemorrhagic septicemia virus IVb isolates and their virulence in northern pike fry

Four viral hemorrhagic septicemia virus (VHSV) genotype IVb isolates were sequenced, their genetic variation explored, and comparative virulence assayed with experimental infections of northern pike Esox lucius fry. In addition to the type strain MI03, the complete 11183 bp genome of the first round goby Neogobius melanostomus isolate from the St. Lawrence River, and the 2013 and 2014 isolates from gizzard shad Dorosoma cepedianum die-offs in Irondequoit Bay, Lake Ontario and Dunkirk Harbor, Lake Erie were all deep sequenced on an Illumina platform. Mutations documented in the 11 yr since the MI03 index case from Lake St. Clair muskellunge Esox masquinongy showed 87 polymorphisms among the 4 isolates. Twenty-six mutations were non-synonymous and located at 18 different positions within the matrix protein, glycoprotein, non-virion protein, and RNA polymerase genes. The same 4 isolates were used to infect northern pike fry by a single 1 h bath exposure. Cumulative percent mortality varied from 42.5 to 62.5%. VHSV was detected in 57% (41/72) of the survivors at the end of the 21-d trial, suggesting that the virus was not rapidly cleared. Lesions were observed in many of the moribund and dead northern pike, such as hemorrhaging in the skin and fins, as well as hydrocephalus. Mean viral load measured from the trunk and visceral tissues of MI03-infected pike was significantly higher than the quantities detected in fish infected with the most recent isolates of genotype IVb, but there were no differences in cumulative mortality observed.

Diseases of Aquatic Organisms

Paramyxoviruses of fish

The first fish paramyxovirus was isolated from normal adult Chinook salmon returning to a coastal hatchery in Oregon in the fall of 1982. Subsequently, the virus was isolated from other stocks of adult Chinook salmon and one stock of adult coho salmon in California, Oregon, Washington and Alaska, leading to its designation as the Pacific salmon paramyxovirus ( PSPV ). The slow-growing virus can be isolated from tissues and ovarian fluids of healthy adult fish returning to spawn and apparently causes no clinical signs of disease or mortality. In 1995, a different and widely disseminated paramyxovirus was isolated from farmed Atlantic salmon in Norway and was designated as Atlantic salmon paramyxovirus ( ASPV ). Although this virus caused no disease or mortality when injected into juvenile Atlantic salmon, ASPV has been associated with proliferative gill inflammation in sea-reared yearling fish; however, additional infectious agents may be involved in the etiology of the condition. Sequence analysis of PSPV and ASPV isolates using the polymerase gene established their placement in the family Paramyxoviridae and has shown the two viruses to be closely related but sufficiently different from each other and from other known paramyxoviruses to possibly represent new genera within the family. The viruses can be diagnosed by isolation in cell culture with final confirmation by molecular methods. Other paramyxovirus-like agents have been observed or isolated from rainbow trout in Germany, from seabream in Japan associated with epithelial necrosis, from turbot in Spain associated with erythrocytic inclusion bodies and buccal/opercular hemorrhaging and from koi and common carp associated with gill necrosis in the European Union.

Book chapter

Hepeviruses of fish

Originally reported from California, the cutthroat trout virus (CTV) has now been isolated from eight species of salmonids in North America. Early work focused on the replication and physical characteristics of the small, round virus, but not until 20 years later was it determined to be most closely related to viruses causing hepatitis E in humans or infecting avian and mammalian hosts. The genome of CTV consists of 7269 nucleotides of positive-sense, single-stranded RNA with a genome organization similar to other members of the family Hepeviridae , although the amino acid sequence identity appears low enough to support creation of a novel genus. While CTV has not been associated with acute disease in fish, the virus was able to form persistently infected cell cultures that may aid research in treatment of hepatitis E-like viruses affecting humans or other animals. Interestingly, trout exposed to CTV were protected for about a month against subsequent exposure to Infectious hematopoietic necrosis virus . Replicating agents suspected to be CTV can be confirmed by polymerase chain reaction (PCR) and sequencing.

Book chapter

Isolation and molecular characterization of a novel picornavirus from baitfish in the USA

During both regulatory and routine surveillance sampling of baitfish from the states of Illinois, Minnesota, Montana, and Wisconsin, USA, isolates (n = 20) of a previously unknown picornavirus were obtained from kidney/spleen or entire viscera of fathead minnows ( Pimephales promelas ) and brassy minnows ( Hybognathus hankinsoni ). Following the appearance of a diffuse cytopathic effect, examination of cell culture supernatant by negative contrast electron microscopy revealed the presence of small, round virus particles (∼30–32 nm), with picornavirus-like morphology. Amplification and sequence analysis of viral RNA identified the agent as a novel member of the Picornaviridae family, tentatively named fathead minnow picornavirus (FHMPV). The full FHMPV genome consisted of 7834 nucleotides. Phylogenetic analysis based on 491 amino acid residues of the 3D gene showed 98.6% to 100% identity among the 20 isolates of FHMPV compared in this study while only 49.5% identity with its nearest neighbor, the bluegill picornavirus (BGPV) isolated from bluegill ( Lepomis macrochirus ). Based on complete polyprotein analysis, the FHMPV shared 58% (P1), 33% (P2) and 43% (P3) amino acid identities with BGPV and shared less than 40% amino acid identity with all other picornaviruses. Hence, we propose the creation of a new genus ( Piscevirus ) within the Picornaviridae family. The impact of FHMPV on the health of fish populations is unknown at present.

Illinois, Minnesota, Montana, Wisconsin