In characteristics of the first North American isolates of viral hemorrhagic septicemia virus
No abstract available
Conference Paper↗
Geology topics
W.N. Batts
Publications and source records attributed to W.N. Batts.
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↗
Isolation of infectious hematopoietic necrosis virus from a leech (Piscicola salmositica) and a copepod (Salmincola sp.), ectoparasites of sockeye salmon Oncorhynchus nerka
Infectious haematopoietic necrosis (IHN) virus was isolated from freshwater leeches Piscicola salmositica and copepods Salmincola sp. removed from the gills of spawning sockeye salmon, Oncorhynchus nerka. This is the first report of the isolation of IHN virus from an animal other than salmonid fishes. High levels of IHN virus were also found in leeches taken from the bottom gravel of the spawning area. The prevalence of IHN virus in samples of individual leeches was as high as 100% and the virus was isolated from 95% of pooled samples of copepod and 1.5 × 10 8 pfu/g in the leech. The level of virus in leeches removed from fish gills was sometimes higher than the level of virus in the gill tissue itself. Virus persisted for at least 16 d in leeches held in the laboratory without feeding. Transmission of IHN virus by leeches probably increases the infection rate of spawning sockeye salmon.
Diseases of Aquatic Organisms↗
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↗
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↗
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↗
Infectious hematopoietic necrosis virus detected by separation and incubation of cells from salmonid cavity fluid.
Infectious hematopoietic necrosis (IHN) virus is usually detected by inoculating susceptible cell cultures with cavity ("ovarian") fluid (CF) from spawning females. We identified additional adult carriers of virus in spawning populations of steelhead trout ( Salmo gairdneri ) and sockeye salmon ( Oncorhynchus nerka ) by collecting nonerythrocytic cells from CF samples by low-speed centrifugation, culturing the cells for at least 7 d at 15 °C, and then testing the culture medium for virus. Virus appeared in the cultured cells from some samples of CF that remained negative during incubation. In additional samples of CF from these species, the virus titer increased in cultured cells compared with the titer in the original CF sample. With chinook salmon ( O . tshawytscha ), no negative samples converted to positive during incubation, but the virus titer was retained in incubated CF cells, but not in cell-free CF.
Canadian Journal of Fisheries and Aquatic Sciences↗
Testing of male sockeye salmon ( Oncorhynchus nerka ) and steelhead trout ( Salmo gairdneri ) for infectious hematopoietic necrosis virus
Infectious hematopoietic necrosis (IHN) virus has been isolated only rarely from whole milt samples of male sockeye salmon ( Oncorhynchus nerka ). In 3 yr of testing, virus incidences in males ranged from 0 to 13% when milt was sampled but were 60–100% with spleen or kidney. When IHN virus was isolated from sockeye salmon milt at titers less than 3.00 log 10 plaque-forming units (pfu)/mL, the level of virus in the kidney or spleen exceeded 7.00 log 10 pfu/g. Higher rates of IHN virus isolation from kidney or spleen than from milt were also generally found in steelhead trout ( Salmo gairdneri ), although the differences were less pronounced than in sockeye salmon. Furthermore, virus was sometimes isolated from steelhead trout milt when the level of virus in kidney or spleen samples was very low, and was recovered from some milt samples when none was isolated from the corresponding spleen sample. When male salmonids are tested for IHN virus, kidney or spleen samples are superior to whole milt, but milt should be included for critical examinations.
Canadian Journal of Fisheries and Aquatic Sciences↗