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The Oregon sockeye salmon virus (IHN) : A. Replication and autointerference. B. The effect of temperature on infection in Kokanee salmon (Oncorhynchus nerka) and on virus stability

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Title The Oregon sockeye salmon virus (IHN) : A. Replication and autointerference. B. The effect of temperature on infection in Kokanee salmon (Oncorhynchus nerka) and on virus stability
Names McAllister, Philip E. (creator)
Pilcher, K. S. (advisor)
Date Issued 1973-05-01 (iso8601)
Note Graduation date: 1973
Abstract Since the isolation of the Oregon sockeye salmon virus (OSV) in
1958, extensive investigations have been undertaken to characterize
the properties of this virus. The results of these investigations
have indicated that OSV is a single-stranded RNA virus which contains
essential lipids and has a density of 1.16 gm/cm³ in sucrose.
In negative stained, glutaraldehyde-fixed preparations, OSV has been
identified as a bullet shaped particle 166 to 181 nm in length and 91
to 98 nm in diameter. OSV has been tentatively placed in the
Rhabdovirus group. OSV has been found to be antigenically indistinguishable
from infectious hematopoietic necrosis (IHN) virus, a
bullet shaped virus of sockeye salmon and rainbow trout.
Events occurring during a single cycle of OSV replication were
investigated. OSV was found to adsorb rapidly to the cell monolayer
in the first 15 minutes of exposure. By 60 to 75 minutes after virus
exposure under the conditions used, adsorption appeared to be
essentially completed.
A single-cycle growth curve of OSV indicated that free and total
virus increase in an exponential manner. Total virus reached maximum
titer in 24 hours while free virus did not reach maximum titer
until 48 to 72 hours after infection. Progeny virus appear to be
retained in the cell and released gradually into the surrounding
medium. The titer attained by free and total virus was nearly the
same.
Total viral RNA synthesis during a single cycle of OSV replication
was monitored by incorporation of uridine-5-³ into acid-precipitable
RNA. Total viral RNA in actinomycin D treated monolayers
increased exponentially from 8 to 17 hours after infection then
stabilized. Accumulation of total viral RNA corresponded well to
the total virus growth curve. RNA incorporation into infectious virus
continued after total viral RNA reached a maximum. Cellular RNA
synthesis was reduced by greater than 99% in the presence of
actinomycin D.
Indirect fluorescent antibody staining indicated that OSV
developed in the cytoplasm. Fluorescent granules appeared at 2 hours after virus infection and increased in size and number on
longer incubation. The fluorescence observed was confined to the
cytoplasm with no fluorescence observable in the nucleus.
Direct evidence for the occurrence of the autointerference
phenomenon with OSV was demonstrated with serial undiluted virus
passage. Serial undiluted virus passage titers were consistently 10
to 100 times lower than parallel titers achieved in serial diluted
virus passage indicating that autointerference occurs with OSV.
Thin section preparations of cells infected with OSV indicated
a morphological difference in virus particles produced using diluted
virus inoculum and undiluted virus inoculum (autointerfering conditions).
Three types of particles were observed. OSV-I particles
were 188 nm in length by 70 nm in diameter; OSV-II particles were
118 by 69 nm; and OSV-III particles were 81 by 66 nm. In cells
exposed to diluted virus inoculum, essentially only OSV-I particles
were observed. In cells exposed to undiluted virus inoculum, all
three types of particles were observed. OSV-III particles were found
in the greatest number while OSV-I particles were found in the least
number.
Negative stained virus preparations gave some insight into the
nature of the OSV nucleocapsid. The striated component of the virus
appears to be helical in configuration. The striations are 2.4 nm in
width with periodicity of 5.2 nm. Situated within and observed
extruding from the striated component, there appears to be filamentous
material which in these preparations appeared to have a random
configuration. It is not known if the striated and filamentous
components are continuous.
Thin section preparations indicated the presence of 12 nm
surface projections extending radially from the surface of the virus
particle.
Virus particles were observed in cytoplasmic vesicles and
were observed budding through surface membranes into the surrounding
medium. No inclusion bodies were observed in either the
cytoplasm or the nucleus. No virus particles were observed in the
nucleus.
The thermal stability of OSV was investigated in three aqueous
environments. The virus became more unstable with increasing
temperature. All virus titers decreased exponentially.
The effect of temperature on OSV infection in kokanee salmon
was investigated. The optimum temperature range for the progress
of the infection was 12.2°C to 15.0°C. At temperatures above
17.8°C, the percent mortality decreased significantly. At temperatures
below 9.4°C, the percent mortality was very high, but the
mean time for death was greatly increased.
The antigenic relationship between OSV and Egtved virus, a
bullet shaped virus of rainbow trout, was investigated by the cross plaque neutralization test. No neutralization of the heterologous
virus was observed with either antiserum. By this method OSV and
Egtved virus appear to be antigenically unrelated.
Genre Thesis/Dissertation
Topic Salmon -- Diseases
Identifier http://hdl.handle.net/1957/45843

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