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W.N. Batts

Publications and source records attributed to W.N. Batts.

At least 19 recordsLinked to original sources
USGS2011

Emergence of viral hemorrhagic septicemia virus in the North American Great Lakes region is associated with low viral genetic diversity

Viral hemorrhagic septicemia virus (VHSV) is a fish rhabdovirus that causes disease in a broad range of marine and freshwater hosts. The known geographic range includes the Northern Atlantic and Pacific Oceans, and recently it has invaded the Great Lakes region of North Ame­rica. The goal of this work was to characterize genetic diversity of Great Lakes VHSV isolates at the early stage of this viral emergence by comparing a partial glycoprotein (G) gene sequence (669 nt) of 108 isolates collected from 2003 to 2009 from 31 species and at 37 sites. Phylogenetic analysis showed that all isolates fell into sub-lineage IVb within the major VHSV genetic group IV. Among these 108 isolates, genetic diversity was low, with a maximum of 1.05% within the 669 nt region. There were 11 unique sequences, designated vcG001 to vcG011. Two dominant sequence types, vcG001 and vcG002, accounted for 90% (97 of 108) of the isolates. The vcG001 isolates were most widespread. We saw no apparent association of sequence type with host or year of isolation, but we did note a spatial pattern, in which vcG002 isolates were more prevalent in the easternmost sub-regions, including inland New York state and the St. Lawrence Seaway. Different sequence types were found among isolates from single disease outbreaks, and mixtures of types were evident within 2 isolates from ­individual fish. Overall, the genetic diversity of VHSV in the Great Lakes region was found to be extremely low, consistent with an introduction of a new virus into a geographic region with ­previously naïve host populations.

Diseases of Aquatic Organisms↗
USGS2008

Genetic analysis of paramyxovirus isolates from pacific salmon reveals two independently co-circulating lineages

Viruses with the morphological and biochemical characteristics of the family Paramyxoviridae (paramyxoviruses) have been isolated from adult salmon returning to rivers along the Pacific coast of North America since 1982. These Pacific salmon paramyxoviruses (PSPV), which have mainly been isolated from Chinook salmon Oncorhynchus tshawytscha, grow slowly in established fish cell lines and have not been associated with disease. Genetic analysis of a 505-base-pair region of the polymerase gene from 47 PsPV isolates produced 17 nucleotide sequence types that could be grouped into two major sublineages, designated A and B. The two independently co-circulating sublineages differed by 12.1-13.9% at the nucleotide level but by only 1.2% at the amino acid level. Isolates of PSPV from adult Pacific salmon returning to rivers from Alaska to California over a 25-year period showed little evidence of geographic or temporal grouping. Phylogenetic analyses revealed that these paramyxoviruses of Pacific salmon were most closely related to the Atlantic salmon paramyxovirus (ASPV) from Norway, having a maximum nucleotide diversity of 26.1 % and an amino acid diversity of 19.0%. When compared with homologous sequences of other paramyxoviruses, PSPV and ASPV were sufficiently distinct to suggest that they are not clearly members of any of the established genera in the family Paramyxoviridae. in the course of this study, a polymerase chain reaction assay was developed that can be used for confirmatory identification of PSPV. ?? Copyright by the American Fisheries Society 2008.

Journal of Aquatic Animal Health↗
USGS2007

Detection of viral hemorrhagic septicemia in round gobies in New York State (USA) waters of Lake Ontario and the St. Lawrence River

In May 2006 a large mortality of several thousand round gobies Neogobius melanostomus (Pallas, 1814) occurred in New York waters of the St. Lawrence River and Lake Ontario. Necropsies of sampled fish from these areas showed pallor of the liver and gills, and hemorrhagic areas in many organs. Histopathologic examination of affected tissues revealed areas of necrosis and hemorrhage. Inoculations of fathead minnow Pimephales promelas (Rafinesque, 1820) cell cultures with dilutions of tissue samples from the necropsied gobies produced a cytopathic effect within 5 d post-inoculation. Samples of cell culture supernatant were tested using RT-PCR and confirmed the presence of viral hemorrhagic septicemia virus (VHSV). Sequence analysis of the VHSV isolate resulted in its assignment to the type-IVb subgroup. The detection of VHSV in a relatively recent invasive fish species in the Great Lakes and the potential impact of VHSV on the ecology and economy of the area will require further investigation and careful management considerations. Inter-Research 2007.

New York↗
USGS2005

Sequence motifs and prokaryotic expression of the reptilian paramyxovirus fusion protein

Fourteen reptilian paramyxovirus isolates were chosen to represent the known extent of genetic diversity among this novel group of viruses. Selected regions of the fusion (F) gene were sequenced, analyzed and compared. The F gene of all isolates contained conserved motifs homologous to those described for other members of the family Paramyxoviridae including: signal peptide, transmembrane domain, furin cleavage site, fusion peptide, N-linked glycosylation sites, and two heptad repeats, the second of which (HRB-LZ) had the characteristics of a leucine zipper. Selected regions of the fusion gene of isolate Gono-GER85 were inserted into a prokaryotic expression system to generate three recombinant protein fragments of various sizes. The longest recombinant protein was cleaved by furin into two fragments of predicted length. Western blot analysis with virus-neutralizing rabbit-antiserum against this isolate demonstrated that only the longest construct reacted with the antiserum. This construct was unique in containing 30 additional C-terminal amino acids that included most of the HRB-LZ. These results indicate that the F genes of reptilian paramyxoviruses contain highly conserved motifs typical of other members of the family and suggest that the HRB-LZ domain of the reptilian paramyxovirus F protein contains a linear antigenic epitope. ?? Springer-Verlag 2005.

Archives of Virology↗
USGS2004

Molecular characterization of birnaviruses isolated from wild marine fishes at the Flemish Cap (Newfoundland)

Several isolates of aquatic birnaviruses were recovered from different species of wild fish caught in the Flemish Cap, a Newfoundland fishery close to the Atlantic coast of Canada. The nucleotide sequence of a region of the NS gene was identical among the isolates and was most similar to the Dry Mills and West Buxton reference strains of infectious pancreatic necrosis virus (IPNV). Phylogenetic analysis of the sequence of a region of the VP2 gene demonstrated that the isolates were most closely aligned with the American strains of IPNV serotype Al. Electron microscopy of virus structures clarified and concentrated from cultures of infected chinook salmon embryo (CHSE-214) cells revealed a majority of typical IPNV-like icosahedral particles, as well as a low proportion of type I tubules having a diameter of approximately 55 nm and a variable length of up to 2 µm . The tubules could be propagated in cell cultures, but always in the presence of low proportions of icosahedral particles. Cloning of selected isolates by serial dilution yielded preparations with a high proportion of the tubular structures with a density in CsCl gradients of approximately 1.30 g cm -3 . Polyacrylamide gel electrophoresis revealed the material in the band was composed of the IPNV pVP2 and VP2 proteins.

Flemish Cap↗
USGS2003

Host and geographic range extensions of the North American strain of viral hemorrhagic septicemia virus

Viral hemorrhagic septicemia virus (VHSV) was isolated from populations of Pacific sardine Sardinops sagax from the coastal waters of Vancouver Island, British Columbia, Canada, and central and southern California, USA. The virus was also isolated from Pacific mackerel Scomber japonicus in southern California, from eulachon or smelt Thaleichthys pacificus , and surf smelt Hypomesus pretiosus pretiosus from Oregon, USA. Mortality and skin lesions typical of viral hemorrhagic septicemia in other marine fish species were observed among sardine in Canada and in a few surf smelt from Oregon, but the remaining isolates of VHSV were obtained from healthy appearing fish. The prevalence of VHSV among groups of apparently healthy sardine, mackerel and smelt ranged from 4 to 8% in California and Oregon. A greater prevalence of infection (58%) occurred in groups of sardine sampled in Canada that sustained a naturally occurring epidemic during 1998-99. A captive group of surf smelt in Oregon exhibited an 81% prevalence of infection with clinical signs in only a few fish. The new isolates were confirmed as North American VHSV and were closely related based on comparisons of the partial nucleotide sequence of the glycoprotein (G) gene. The VHSV isolates from sardine in Canada and California were the most closely related, differing from isolates obtained from other marine fish species and salmonids in British Columbia, Canada, Alaska and Washington, USA. These new virus isolations extend both the known hosts (sardine, mackerel and 2 species of smelt) and geographic range (Oregon and California, USA) of VHSV.

Diseases of Aquatic Organisms↗
USGS2001

Negligible risk associated with the movement of processed rainbow trout, Oncorhynchus mykiss (Walbaum), from an infectious haematopoietic necrosis virus (IHNV) endemic area

To assess the risk of transmission of infectious haematopoietic necrosis virus (IHNV) associated with the movement of processed rainbow trout, Oncorhynchus mykiss, from an area where the virus is endemic, 240 freshly eviscerated fish (225-500 g) exhibiting spinal curvature or spinal compression types of deformities were tested for IHNV by virus isolation and polymerase chain reaction (PCR) techniques. Commercially produced rainbow trout, approximately 1-year-old, that exhibited spinal deformities were considered to have had a high likelihood of having survived an outbreak of IHN. Serological analysis of fish exhibiting spinal curvature or spinal compression types of deformities for anti-IHNV antibodies resulted, in 71 and 50% of the serum samples, respectively, with detectable neutralization activity suggesting previous infection with IHNV. A portion of the skin and muscle in the area of the deformity was collected, as well as brain tissue from each commercially processed fish. Tissue homogenates were tested for IHNV using the epithelioma papulosum cyprini (EPC) cell line pretreated with polyethylene glycol and the chinook salmon embryo (CHSE-214) cell line using standard methods. Nested, reverse transcriptase (RT)-PCR for the detection of IHNV used the central 1231 bp portion of the glycoprotein (G) challenge studies and is suggested as a mechanism responsible for virus clearance. These results provide scientific information that can be used to assess the risk associated with the movement of processed rainbow trout from an IHNV endemic area.

Journal of Fish Diseases↗
USGS1999

Comparative sequence analyses of sixteen reptilian paramyxoviruses

Viral genomic RNA of Fer-de-Lance virus (FDLV), a paramyxovirus highly pathogenic for reptiles, was reverse transcribed and cloned. Plasmids with significant sequence similarities to the hemagglutinin-neuraminidase (HN) and polymerase (L) genes of mammalian paramyxoviruses were identified by BLAST search. Partial sequences of the FDLV genes were used to design primers for amplification by nested polymerase chain reaction (PCR) and sequencing of 518-bp L gene and 352-bp HN gene fragments from a collection of 15 previously uncharacterized reptilian paramyxoviruses. Phylogenetic analyses of the partial L and HN sequences produced similar trees in which there were two distinct subgroups of isolates that were supported with maximum bootstrap values, and several intermediate isolates. Within each subgroup the nucleotide divergence values were less than 2.5%, while the divergence between the two subgroups was 20-22%. This indicated that the two subgroups represent distinct virus species containing multiple virus strains. The five intermediate isolates had nucleotide divergence values of 11-20% and may represent additional distinct species. In addition to establishing diversity among reptilian paramyxoviruses, the phylogenetic groupings showed some correlation with geographic location, and clearly demonstrated a low level of host species-specificity within these viruses. Copyright (C) 1999 Elsevier Science B.V.

Virus Research↗
USGS1994

Association of viral hemorrhagic septicemia virus with epizootic hemorrhages of the skin in Pacific herring Clupea harengus pallasi from Prince William Sound and Kodiak Island, Alaska, USA

Only one-third of the Pacific hernng Clupea harengus pallasi expected to spawn in Prince William Sound (PWS), Alaska, USA, in sprlng 1993 were observed. Of these herring, 15 to 43 '% had external ulcers or subdermal hemorrhages of the skln and fins. A rhabdovirus identified as the North American strain of viral hemorrhagic septicemia virus (VHSV) was isolated from affected herring and 1 Pacific cod Gadus macrocephalus with skin lesions from PWS, and from herring with similar lesions collected near Kodiak Island. No other pathogens were detected in the herring examined. Although VHSV may have been responsible for the skin lesions, there was no confirmed mass herring mortality observed in PWS; hence the actual cause of the reduced herring numbers is still unknown. The same strain of VHSV was subsequently isolated from captive juvenile herring collected from Auke Bay, Alaska, near Juneau, from herring in British Columbia, Canada, and from Puget Sound, Washington, USA. These findings suggest the virus is an opportunistic pathogen that is widely indigenous to Pacific herring populations in the Pacific Northwest and that herring are a significant marine reservoir for North American VHSV.

Alaska↗
USGS1992

In vitro infection of salmonid epidermal tissues by infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus

The ability of two rhabdoviruses, infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV), to infect fish skin was investigated by in vitro infection of excised tissues. Virus replication was determined by plaque assay of homogenized tissue extracts, and the virus antigen was detected by immunohistology of tissue sections. Gill, fin, and ventral abdominal skin tissues of rainbow trout Oncorhynchus mykiss that had been infected in vitro with a virulent strain of IHNV (193–110) produced substantial increases in virus titer within 24 h. Titers continued to increase up until day 3 of incubation; by this time, virus had increased 1,000-fold or more. This increase in IHNV titer occurred in epidermal tissues of fingerlings and of older fish. In another experiment, IHNV replicated in excised rainbow trout tissues whether the fish had been subject to prior infection with a virulent strain of IHNV (Western Regional Aquaculture Consortium isolate) or whether the fish had been infected previously with an attenuated strain of the virus (Nan Scott Lake, with 100 passes in culture). A virulent strain of VHSV (23/75) replicated effectively in excised gill tissues and epidermal tissues of rainbow trout and chinook salmon O. tshawytscha ; however, the avirulent North American strain of VHSV (Makah) replicated poorly or not at all.

Journal of Aquatic Animal Health↗
USGS1992

Identification of viral hemorrhagic septicemia virus isolated from Pacific cod Gadus macrocephalus in Prince William Sound Alaska, USA

Ulcerative slun tissues from 2 Pacific cod Gadus rnacrocephalus caught in Prince William Sound, Alaska, USA, were examined for virus by Fish Pathology staff within the F.R.E.D. Division of the Alaska Department of Fish and Game. Six days after inoculation of Epitheliorna papulosum cyprini (EPC) cells at 14"C, diffuse rounding and lifting of cells from the monolayers suggestive of cytopathlc effect became visible in the lower sample dilutions. Ultrastructural examinations of affected EPC cells showed rhabdovirus particles within cytoplasmic vacuoles and on the cell surface membranes. Virus isolates from both cod were subsequently confirmed as viral hemorrhagic septicemia virus (VHSV) by serum neutralizabon and immunoblot assay. This is the first VHSV isolated from Pacific cod, which represents a new host species for the virus. Histologically, cod skin ulcers appeared to be caused by a foreign-body-type inflammatory response to foci of protozoa resembling X cells that also had plasmodial stages. Whether the rhabdovirus was incidental to the slun lesion or played a role in its etiology remains to be determined. The possible relationship between thls virus and the recent occurrences of VHSV in anadromous salmoruds from Washington State, USA, is discussed.

Diseases of Aquatic Organisms↗
USGS1990

Multiplication of infectious hematopoietic necrosis virus in rainbow trout following immersion infection: whole-body assay and immunohistochemistry

The sites of replication of infectious hematopoietic necrosis virus (IHNV) in infected tissues were detected in fingerling rainbow trout Oncorhynchus mykiss by in situ histologic techniques following immersion infection. Virus antigens in tissues were detected by a neutralizing mouse monoclonal antibody and a one-step anti-mouse biotin-streptavidin conjugated to horseradish peroxidase. The efficiency of infection and virulence of the virus determined by mortality rates showed high virulence of the selected IHNV isolates, and viral replication in individual fish showed that virus content of the fish increased rapidly from the second day to the seventh day postinfection. The earliest viral lesions following infection were detected in the epidermis of the pectoral fins, opercula, and ventral surface of the body. Virus lesions became evident in kidneys on the third day. By the fifth day, when there was a significant increase in virus titer, foci of viral replication were detected in gill tissue and in the anterior internal tissues below the epidermis. Subsequently, extensive virus replication and tissue destruction were observed in the spleen, dorsal adipose tissues, ventricle, and pseudobranch. Replication in the liver, the muscularis layers of the digestive tract, and the general body musculature followed later. These infection experiments indicated that the epidermis and gills of fish constitute important sites of early IHNV replication.

Journal of Aquatic Animal Health↗
USGS1989

Concentration of infectious hematopoietic necrosis virus from water samples by tangential flow filtration and polyethylene glycol precipitation

Infectious hematopoietic necrosis virus (IHNV) was concentrated from water samples by polyethylene glycol (PEG) precipitation, tangential flow filtration (TFF), and by a combination of TFF followed by PEG precipitation of the retentate. Used alone, PEG increased virus titers more than 200-fold, and the efficiency of recovery was as great as 100%. Used alone, TFF concentrated IHNV more than 20-fold, and average recovery was 70%. When the two techniques were combined, 10-L water samples were reduced to about 300 mL by TFF and the virus was precipitated with PEG into a 1 to 2 g pellet; total recovery was as great as 100%. The combined techniques were used to isolate IHNV from water samples taken from a river containing adult sockeye salmon ( Oncorhynchus nerka ) and from a hatchery pond containing adult spring chinook salmon ( O . tshawytscha ). The combination of these methods was effective in concentrating and detecting IHNV from water containing only three infectious particles per 10-L sample.

Canadian Journal of Fisheries and Aquatic Sciences↗
USGS1989

Enhanced detection of infectious hematopoietic necrosis virus by pretreatment of cell monolayers with polyethylene glycol

To improve quantification of very low levels of infectious hematopoietic necrosis virus (IHNV) in samples of tissue, ovarian fluid, or natural water supplies, we tested the ability of polyethylene glycol (PEG) to enhance the sensitivity and speed of the plaque assay system. We compared 4, 7, and 10% solutions of PEG of molecular weight 6,000, 8,000, or 20,000 applied at selected volumes and for various durations. When cell monolayers of epithelioma papulosum cyprini (EPC), fathead minnow (FHM), chinook salmon embryo (CHSE-214), and bluegill fry (BF2) were pretreated with 7% PEG-20,000, they produced 4-17-fold increases in plaque assay titers of IHNV. The plaque assay titers of viral hemorrhagic septicemia virus, chum salmon reovirus, and chinook salmon paramyxovirus were also enhanced by exposure of CHSE-214 cells to PEG, but the titers of infectious pancreatic necrosis virus and Oncorhynchus masou virus were not substantially changed. Plaques formed by IHNV on PEG-treated EPC cells incubated at 15°C had a larger mean diameter at 6 d than those on control cells at 8 d; this suggests the assay could be shortened by use of PEG. Pretreatment of EPC cell monolayers with PEG enabled detection of IHNV in some samples that appeared negative with untreated cells. For example, when ovarian fluid samples from chinook salmon Oncorhynchus tshawytscha were inoculated onto untreated monolayers of EPC cells, IHNV was detected in only 11 of 51 samples; 17 of the samples were positive when PEG-treated EPC cells were used. PDF

Journal of Aquatic Animal Health↗
USGS1989

Comparison of infectious hematopoietic necrosis in natural and experimental infections of spawning salmonids by infectivity and immunohistochemistry

Infectious hematopoietic necrosis (IHN) continues to be a serious virus disease of salmonids with epizootics recorded in both wild and hatchery populations (Williams and Amend 1976; Carlisle et al 1979; Groberg and Fryer 1983; Saft and Pratt 1986; Traxler 1987; Follett et al 1987; Meyers et al 1988). While originally enzootic in western North America, the virus appears to be spreading further (Sano et al 1977; de Kinkelin et al 1987; Bovo et al 1987). In hatchery outbreaks occurring in regions where the virus is not enzootic, it is often possible to trace the virus to the importation of infected fingerlings or contaminated eggs. In regions where the virus is widespread among stocks of fish, the source of virus infection is more difficult to establish particularly in watersheds where there are anadromous salmonids. Although salmonid fish surviving infection as fry and returning from the ocean to spawn are considered to be parental carriers of IHNV, there is very little data to support this hypothesis. Amend (1975) in the study of rainbow trout reported that in a population surviving infection and assayed a few years later found that a few trout were carrying virus. This is the study often cited as evidence for the carrier status of returning salmonids. LaPatra et al (1987) presented data that indicated IHNV has been transmitted horizontally through water from adult chinook salmon (Oncorhynchus tshawytscha) to adult coho salmon (O. kisutch) at a hatchery in northern California. They suggested that horizontal transmission may be an important means for perpetuating IHN. However, the actual mechanisms for persistence and transmission of IHN among fish in a watershed is likely to be complex and involve multiple species and age groups as well as intermediate vectors and/or reservoirs.

Book chapter↗