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Geology topics

J. R. Winton

Publications and source records attributed to J. R. Winton.

At least 55 records · Page 3Linked to original sources
USGS2001

Epidemiology of viral hemorrhagic septicemia (VHS) among juvenile Pacific herring and Pacific sandlances in Puget Sound, Washington

Viral hemorrhagic septicemia (VHS) and the associated virus (VHSV) were identified in newly metamorphosed Pacific herring Clupea pallasi and Pacific sand lances Ammodytes hexapterus captured from Puget Sound, Washington, between 1995 and 1998. During that 4-year period, virus was detected in less than 1% of free-ranging, age-0 Pacific herring; however, when groups of these fish were confined in the laboratory, they experienced severe mortality, occasionally exceeding 50%, with the prevalence of VHSV reaching 100% by 14 d postcapture. At 7–21 d postcapture, VHSV titers peaked in excess of 108 plaque-forming units/g of tissue; by 30 d postcapture, however, the virus could no longer be isolated. Fish surviving beyond 30 d eliminated the virus from their tissues, but some remained lethargic and continued to show signs of hemorrhage around the mouth, skin, and fins until about 6 weeks postcapture. No cutaneous ulcers were observed during either the acute or the recovery phases of infection. Eighteen-month-old Pacific herring captured from the same area were also negative for VHSV but developed active infections after confinement for 7 d. Unlike younger fish, only 8.4% of these older fish died of VHS, and 7.7% of survivors were positive for VHSV at 7–10 d postcapture, which suggests that a higher proportion of the older fish had developed resistance to VHSV from prior exposure to it. Three months after fatalities ceased in the laboratory-held fish, the surviving fish were challenged with 5 3 103 plaque-forming units/mL of VHSV for 1 h. No mortality was observed during the next 30 d, and virus was recoverable at very low titers in fewer than 5% of the challenged fish, indicating the development of an active immunity to VHSV. Laboratory cohabitation of infected wild Pacific herring with laboratory-reared, specific-pathogen-free Pacific herring resulted in transmission of VHSV to the nonimmune fish, with the resulting course of disease resembling that seen in wild Pacific herring confined in the laboratory. The possible effects of VHS on stocks of Pacific herring are discussed.

Journal of Aquatic Animal Health↗
USGS2000

Tissue-specific induction of Hsp90 mRNA and plasma cortisol response in chinook salmon following heat shock, seawater challenge, and handling challenge

In studying the whole-body response of chinook salmon ( Oncorhynchus tshawytscha ) to various stressors, we found that 5-hour exposure to elevated temperature (mean 21.6°C; + 10.6°C over ambient) induced a marked increase in Hsp90 messenger RNA accumulation in heart, brain, gill, muscle, liver, kidney, and tail fin tissues. The most vital tissues (heart, brain, gill, and muscle) showed the greatest Hsp90-mRNA response, with heart tissue increasing approximately 35-fold. Heat shock induced no increase in plasma cortisol. In contrast, a standard handling challenge induced high plasma cortisol levels, but no elevation in Hsp90 mRNA in any tissue, clearly separating the physiological and cellular stress responses. We saw no increase either in tissue Hsp90 mRNA levels or in plasma cortisol concentrations after exposing the fish to seawater overnight.

Marine Biotechnology↗
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↗
USGS1999

Successful aquatic animal disease emergency programmes

The authors provide examples of emergency programmes which have been successful in eradicating or controlling certain diseases of aquatic animals. The paper is divided into four parts. The first part describes the initial isolation of viral haemorrhagic septicaemia (VHS) virus in North America in the autumn of 1988 from feral adult chinook ( Oncorhynchus tshawytscha ) and coho salmon ( O.kisutch ) returning for spawning. The fish disease control policies at both State and Federal levels in the United States of America required quarantine and emergency eradication measures upon the finding of certain exotic fish pathogens, including VHS virus. The procedures for emergency plans, destruction of stocks and disinfection of facilities are described, as well as challenge experiments with the North American strains of VHS virus and the detection of the virus in marine fish species (cod [ Gadus macrocephalus ] and herring [ Clupea harengus pallasi ]) in the Pacific Ocean. The second part of the paper outlines the aquatic animal legislation in Great Britain and within the European Union, in regard to contingency plans, initial investigations, action on the suspicion of notifiable disease and action on confirmation of infection. The legal description is followed by an account of an outbreak of viral haemorrhagic septicaemia in turbot (Scophthalmus maximus) in Great Britain, including the stamping-out process at the affected farm and investigations conducted to screen other farms in the vicinity for possible infection. The third part provides a historical review of the build-up of infectious salmon anaemia (ISA) in Norway and the attempts to control the disease using legal measures in the absence of detailed knowledge of the aetiology, epizootiology, pathogenesis, etc. of the disease. The measures taken show that the spread of ISA can be controlled using restrictions on the movement of fish, disinfection procedures, etc. However, acceptance and understanding of the chosen strategy by the fish farmers is a pre-requisite to reach that goal. Finally, the paper summarises future needs for national and international legislation, including the development of standard approaches for control, the creation of appropriate infrastructures and a better understanding of the epidemiology of aquatic animal diseases.

Revue Scientifique et Technique↗
USGS1999

Relative virulence of three isolates of Piscirickettsia salmonis for coho salmon Oncorhynchus kisutch

Piscirickettsia salmonis was first recognized as the cause of mortality among pen-reared coho salmon Oncorhynchus kisutch in Chile. Since the initial isolation of this intracellular Gram-negative bacterium in 1989, similar organisms have been described from several areas of the world, but the associated outbreaks were not reported to be as serious as those that occurred in Chile. To determine if this was due to differences in virulence among isolates of P. salmonis , we conducted an experiment comparing isolates from Chile, British Columbia, Canada, and Norway (LF-89, ATL-4-91 and NOR-92, respectively). For each of the isolates, 3 replicates of 30 coho salmon were injected intraperitoneally with each of 3 concentrations of the bacterium. Negative control fish were injected with MEM-10. Mortalities were collected daily for 41 d post-injection. Piscirickettsiosis was observed in fish injected with each of the 3 isolates, and for each isolate, cumulative mortality was directly related to the concentration of bacterial cells administered. The LF-89 isolate was the most virulent, with losses reaching 97% in the 3 replicates injected with 105.0 TCID50, 91% in the replicates injected with 104.0 TCID50, and 57% in the fish injected with 103.0 TCID50. The ATL-4-91 isolate caused losses of 92% in the 3 replicates injected with 105.0 TCID50, 76% in the fish injected with 104.0 TCID50, and 32% in those injected with 103.0 TCID50. The NOR-92 isolate was the least virulent, causing 41% mortality in the replicates injected with 104.6 TCID50. At 41 d post-injection, 6% of the fish injected with 103.6 TCID50 NOR-92 had died. Mortality was only 2% in the fish injected with 102.6 TCID50 NOR-92, which was the same as the negative control group. Because the group injected with the highest concentration (104.6 TCID50) of NOR-92 was still experiencing mortality at 41 d, it was held for an additional 46 d. At 87 d post-injection, the cumulative mortality in this group had reached 70%. These differences in virulence among the isolates were statistically significant (p < 0.0001), and are important for the management of affected stocks of fish.

Diseases of Aquatic Organisms↗
USGS1999

Epizootiology of viral hemorrhagic septicemia virus in Pacific herring from the spawn-on-kelp fishery in Prince William Sound, Alaska, USA

Both the prevalence and tissue titer of viral hemorrhagic septicemia virus (VHSV) increased in Pacific herring Clupea pallasi following their introduction into net pens (pounds) used in the closed pound spawn-on-kelp (SOK) fishery in Prince William Sound, Alaska. VHSV was also found in water samples from inside and outside the SOK pounds after herring had been confined for several days; however, water samples taken near wild free-ranging, spawning herring either failed to test positive or tested weakly positive for virus. Little or no virus was found in tissue samples from free-ranging, spawning herring captured from the vicinity of the pounds, nor did the prevalence of VHSV increase following spawning as it did in impounded herring. The data indicated that increased prevalences of VHSV were correlated with confinement of herring for the closed pound SOK fishery and that infection was spread within the pounds through waterborne exposure to virus particles originating from impounded fish. In addition, pounds containing predominantly young fish had higher prevalences of VHSV, suggesting that older fish may be partially immune, perhaps as a result of previous infection with the virus. Operation of SOK pounds during spawning seasons in which young herring predominate may amplify the disease and possibly exacerbate the population fluctuations observed in wild herring stocks.

Diseases of Aquatic Organisms↗
USGS1999

Antigenic and functional characterization of p57 produced by Renibacterium salmoninarum

Renibacterium salmoninarum , the causative agent of bacterial kidney disease, produces large quantities of a 57-58 kDa protein (p57) during growth in broth culture and during infection of salmonid fish. Biological activities of secreted p57 include agglutination of salrnonid leucocytes and rabbit erythrocytes. We define the location of epitopes on p57 recognized by agglutination-blocking monoclonal antibodies (MAbs) 4Cl1, 4H8 and 4D3, and demonstrate that the majority of secreted p57 is a nlonomer that retains salrnonid leucocyte agglutinat~ng activity. The 3 MAbs bound a recombinant, amino-terminal fragment of p57 (211 aa) but not a carboxy-terminal fragment (315 aa) demonstrating that the neutralizing epitopes are located within the amino-terminal portion of p57. When combinations of the MAbs were used in an antigen capture ELISA. the epitopes recognized by the 3 MAbs were shown to be sterically separate. However, when the same MAb was used as both the coating and detection MAb, binding of the biotinylated detection MAb was not observed. These data indicate that the epitopes recognized by the 3 agglutination-blocking antibodies are functionally available only once per molecule and that native p57 exists as a monomer Similar ELISA results were obtained when kidney tissues from 3 naturally infected chinook salmon were assayed. Finally, a p57 monomer was purified using anion exchange and size exclusion chromatography that retained in vitro agglutinating activity. A model in which p57 is released from R. salmoninarum as a biologically active monomer during infection of salmonid fish is proposed.

Diseases of Aquatic Organisms↗
USGS1998

Molecular approaches to fish vaccines

For more than 50 years, researchers have tested a variety of killed, attenuated, and subunit preparations for control offish diseases. The earliest fish vaccines used killed preparations containing whole bacteria, viruses, or parasites and today, several bacterins have become commercially successful with more expected as improved delivery systems and adjuvants are realized. Live, attenuated vaccines have been developed by serial passage of a pathogen in culture or by using naturally occurring mutants and cross-reacting strains. These generally offer excellent protection and are cost-effective, but concerns about residual virulence or their effects on other aquatic species make them difficult candidates for licensing. In recent years, the tools of molecular biology have been applied to construction of a variety of recombinant, engineered, or subunit vaccines for fish. Among the approaches to be discussed are: attenuated strains of viruses and bacteria created by deletion of specific genes associated with virulence, in vitro synthesis of protective antigens from genes cloned into E. coli or baculovirus expression systems, chemical synthesis of peptides that represent protective epitopes, and direct immunization with DNA coding for protective antigens. Preparations representing each of these approaches have been tested in laboratory or field trials with various results and such vaccines promise to be safe and relatively inexpensive if they are able to provide protection when delivered by immersion. A significant advantage of genetically engineered vaccines is the ability to construct multivalent preparations that can protect fish against several pathogens or different strains of the same pathogen. While many of these novel vaccine strategies have been effective at stimulating specific immunity in the laboratory, more work is needed to develop better delivery systems and to overcome potential regulatory concerns.

Journal of Applied Ichthyology↗
USGS1998

Vaccination of rainbow trout against infectious hematopoietic necrosis (IHN) by using attenuated mutants selected by neutralizing monoclonal antibodies

A neutralizing monoclonal antibody against infectious hematopoietic necrosis virus (IHNV) was used to select neutralization-resistant mutants from isolates of virus obtained from adult steelhead Oncorhynchus mykiss returning to the Round Butte Hatchery (RB mutants) on the Deschutes River in Oregon, USA, and from rainbow trout (nonanadromous O. mykiss) at a commercial hatchery in the Hagerman Valley of Idaho, USA (193-110 mutants). Two of the mutants, RB-1 and 193-110-4, were significantly (P < 0.001) attenuated compared with parental strains. Vaccination of rainbow trout by waterborne exposure to the mutants conferred solid protection against challenge with wild-type virus. In some trials, fish vaccinated with the RB-1 mutant at 50% tissue culture infectious doses (TCID50) of 1 X 104-1 x 105 TCID50/mL or with the 193-110-4 mutant at 1 x 102-1 x 103 TCID50/mL, held for 14 d, then challenged with the homologous wild-type strain at 1 x 105 TCID50/mL showed relative percent survival of 95-100% (P < 0.005). There was no significant difference (P > 0.05) in protection among fish exposed to the RB-1 vaccine strain at a dose of 1 x 105 TCID50/mL for periods of either 1, 12, or 24 h, held for 14 d, and then challenged with the wild-type RB isolate, although the 1-h exposure seemed to be somewhat less effective. Fish were vaccinated with the RB-1 strain at 1 x 103-1 x 105 TCID50/mL for 24 h then challenged after 1, 7, 14, or 21 d with the wild-type RB isolate. No significant (P > 0.1) protection was observed at 1 d postvaccination, but the relative percent survival increased progressively at each subsequent challenge period, becoming statistically significant by day 7 (P < 0.001) and beyond. These results suggested that resistance to challenge with wild-type virus resulted from development of IHNV-specific immunity and not from viral interference or interferon induction, and they reinforce the potential of an attenuated vaccine to control this important disease.

Journal of Aquatic Animal Health↗
USGS1998

Correlation between plasma component levels of cultured fish and resistance to bacterial infection

Mortalities of yellowtail Seriola quinqueradiata artificially infected with Lactococcus garvieae and of rainbow trout Oncorhynchus mykiss artificially infected with Vibrio anguillarum were compared with the levels of plasma components measured prior to challenge. The levels of plasma total cholesterol, free cholesterol and phospholipid of fish surviving infection were significantly higher in both yellowtail and rainbow trout than those of fish which died during the challenge test. Mortality of yellowtail with plasma total cholesterol levels lower than 250 mg/100 ml was significantly higher than that of fish which had cholesterol levels higher than 275 mg/100 ml (p < 0.05). Rainbow trout whose cholesterol was lower than 520 mg/100 ml suffered a significantly higher mortality due to vibriosis than fish having cholesterol levels higher than 560 mg/100 ml (p < 0.005). These results indicate that low levels of plasma lipid components may be an indicator of lowered disease resistance in cultured fish.

Fish Pathology↗
USGS1998

Purification of Piscirickettsia salmonis and partial characterisation of antigens

Piscirickettsia salmonis is the etiological agent of salmonid rickettsial septicemia, an economically significant disease affecting the salmon aquaculture industry. As with other rickettsial pathogens, antigenic analysis of P. salmonis has been limited by the inherent difficulties of purifying an intracellular organism away from host cell material. In this report, we describe the use of diatrizoate meglumine and diatrizoate sodium (DMDS) density gradient centrifugation to purify P. salmonis grown in chinook salmon embryo (CHSE-214) cells. Plaque assay titers and total protein assays confirmed that viable P. salmonis was consistently concentrated in a visible band within the DMDS density gradient at a density of 1.15 to 1.16 g ml -1 . Recovery of purified, viable organisms from DMDS density gradients varied from 0.6 to 3%. Preparations of uninfected CHSE-214 cells, CHSE-214 cells infected with P. salmonis , and gradient-purified P. salmonis were compared using sodium dodecyl sulfate polyacrylamide gel electrophoresis to assess the degree of purification and to identify P. salmonis -specific proteins. Although gradient-purified P. salmonis preparations were not completely free of host cell material, 8 bacterial proteins were identified. Polyclonal rabbit antiserum was used in an immunoblot of proteins from purified P. salmonis to identify 3 major and 5 minor antigens. The major antigens of 56, 30 and 20 kDa were potential candidates for experimental vaccines and development of novel diagnostic assays.

Diseases of Aquatic Organisms↗
USGS1998

A ribonuclease protection assay can distinguish spring viremia of carp virus from pike fry rhabdovirus

Thirteen rhabdovirus isolates from 10 teleost fish species as well as reference strains of spring viraemia of carp virus (SVCV) and pike fry rhabdovirus (PFRV) cross-reacted in an indirect immunofluorescence assay and were thus indistinguishable by this method. A ribonuclease protection assay (RPA) using a super(32)P-labeled RNA probe made from a cloned copy of the full length SVCV glycoprotein (G) gene was able to discriminate clearly between the type strains of SVCV and PFRV and among the 13 rhabdovirus isolates. Results for the RPA were generally in agreement with standard serum neutralisation assays; however, the RPA was also able to detect genomic differences between isolates of SVCV. These results have implications for fish disease control programs for SVCV.

Bulletin of the European Association of Fish Patho↗
USGS1997

Sequence variation of the glycoprotein gene identifies three distinct lineages within field isolates of viral hemorrhagic septicemia virus, a fish rhabdovirus

To evaluate the genetic diversity of viral haemorrhagic septicaemia virus (VHSV), the sequence of the glycoprotein genes (G) of 11 North American and European isolates were determined. Comparison with the G protein of representative members of the family Rhabdoviridae suggested that VHSV was a different virus species from infectious haemorrhagic necrosis virus (IHNV) and Hirame rhabdovirus (HIRRV). At a higher taxonomic level, VHSV, IHNV and HIRRV formed a group which was genetically closest to the genus Lyssavirus. Compared with each other, the G genes of VHSV displayed a dissimilar overall genetic diversity which correlated with differences in geographical origin. The multiple sequence alignment of the complete G protein, showed that the divergent positions were not uniformly distributed along the sequence. A central region (amino acid position 245-300) accumulated substitutions and appeared to be highly variable. The genetic heterogeneity within a single isolate was high, with an apparent internal mutation frequency of 1.2 x 10(-3) per nucleotide site, attesting the quasispecies nature of the viral population. The phylogeny separated VHSV strains according to the major geographical area of isolation: genotype I for continental Europe, genotype II for the British Isles, and genotype III for North America. Isolates from continental Europe exhibited the highest genetic variability, with sub-groups correlated partially with the serological classification. Neither neutralizing polyclonal sera, nor monoclonal antibodies, were able to discriminate between the genotypes. The overall structure of the phylogenetic tree suggests that VHSV genetic diversity and evolution fit within the model of random change and positive selection operating on quasispecies.

Journal of General Virology↗
USGS1997

Sequence analysis and expression of the M1 and M2 matrix protein genes of hirame rhabdovirus (HIRRV)

We have cloned and sequenced a 2318 nucleotide region of the genomic RNA of hirame rhabdovirus (HIRRV), an important viral pathogen of Japanese flounder Paralichthys olivaceus. This region comprises approximately two-thirds of the 3' end of the nucleocapsid protein (N) gene and the complete matrix protein (M1 and M2) genes with the associated intergenic regions. The partial N gene sequence was 812 nucleotides in length with an open reading frame (ORF) that encoded the carboxyl-terminal 250 amino acids of the N protein. The M1 and M2 genes were 771 and 700 nucleotides in length, respectively, with ORFs encoding proteins of 227 and 193 amino acids. The M1 gene sequence contained an additional small ORF that could encode a highly basic, arginine-rich protein of 25 amino acids. Comparisons of the N, M1, and M2 gene sequences of HIRRV with the corresponding sequences of the fish rhabdoviruses, infectious hematopoietic necrosis virus (IHNV) or viral hemorrhagic septicemia virus (VHSV) indicated that HIRRV was more closely related to IHNV than to VHSV, but was clearly distinct from either. The putative consensus gene termination sequence for IHNV and VHSV, AGAYAG(A)(7), was present in the N-M1, M1-M2, and M2-G intergenic regions of HIRRV as were the putative transcription initiation sequences YGGCAC and AACA. An Escherichia coli expression system was used to produce recombinant proteins from the M1 and M2 genes of HIRRV. These were the same size as the authentic M1 and M2 proteins and reacted with anti-HIRRV rabbit serum in western blots. These reagents can be used for further study of the fish immune response and to test novel control methods.

Diseases of Aquatic Organisms↗