Geology ReportsSearch

Geology topics

C.K. Arakawa

Publications and source records attributed to C.K. Arakawa.

7 recordsLinked to original sources

Genetic comparison of viral hemorrhagic septicemia virus isolates from North America and Europe

Viral hemorrhagic septicemia virus (VHSV) is the cdusative agent of a serious rhabdoviral d~sease of rainbow trout Oncorhynchus myklss in Europe The first isolation of the vlrus in North Amenca occurred In the fall of 1988 when it was recovered from adult chinook 0 tshawytscha and coho 0 klsutch salmon returning to 2 hatcher~es in the state of Washington, USA The following year, VHSV was isolated from adult coho salmon at 2 other hatcher~es in northwestern Washington In 1990 and 1991, VHSV was recovered from Pacific cod Gadus macrocephalus caught in Pnnce Willlam Sound, Alaska Genetic vanation among the 4 isolates from salmon and the 1990 ~solate from Pacific cod was determ~ned uslng T1 nbonuclease finqerprlnt~ng In addition, 4 d~verse isolates from Europe were lncluded for companson The North Amencan isolates of VHSV formed a slngle fingerprint group In which the 4 isolates from salmonids were h~ghly similar to each other and the isolate from Pacific cod was related but less s~milar The 4 European ~solates which included an isolate from Atlantic cod G morhua, formed a second fingerpnnt group The genetic d~vers~ty among the isolates within each fingerpnnt group was estimated to be less than 5 % whlle the North Amencan and European strains of the virus were judged to differ by more than 5% The results indicate that the North Amerlcan isolates of VHSV are not of European ongln and that the virus may be enzootic wlthin the manne environment.

Diseases of Aquatic Organisms

Development of a biotinylated DNA probe for detection of infectious hematopoietic necrosis virus

A nonrad~oact~ve DNA probe assay was developed to detect and ~dent~fy infect~ous hernatopoiet~c necrosls virus (IHNV) uslng a dot blot format The probe a synthet~c DNA oligonucleot~de labeled enzymatlcally w~th biotln hybnd~zed spec~f~cally w~th nucleocaps~d mRNA extracted from Infected cells early In the vlrus repl~cation cycle A rap~d guan~dln~um th~ocyanate based RNA extraction method uslng RNAzol B and rn~crocentrifuge tubes eff~c~ently pioduced h~gh qual~ty RNA from 3 commonly used f~sh cell llnes, CHSE-214, CHH-1, and EPC The probe reacted with 6 d~verse ~solates of IHNV, but d~d not react \nth 2 related rhabdovlruses of fish viral hemorrhagic septlcemla vlrus and H~rame rhabdovlrus The b~ot~nylated probe was sensltlve detect~ng plcogram levels of target mRNA Detect~on and ~dentif~cat~on of IHNV requ~red 2d when cells wele lnoculated at n~ultiplic~t~es of infect~on (MOI) greater than 2 Flve days were necessary to detect and identify IHNV In cells lnoculated at a MO1 of 0 0002

Diseases of Aquatic Organisms

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

Polymerase chain reaction (PCR) amplification of a nucleoprotein gene sequence of infectious hematopoietic necrosis virus

The polymerase chain reaction [PCR) was used to amplify a portion of the nucleoprotein [NI gene of infectious hematopoietic necrosis virus (IHNV). Using a published sequence for the Round Butte isolate of IHNV, a pair of PCR pnmers was synthesized that spanned a 252 nucleotide region of the N gene from residue 319 to residue 570 of the open reading frame. This region included a 30 nucleotide target sequence for a synthetic oligonucleotide probe developed for detection of IHNV N gene messenger RNA. After 25 cycles of amplification of either messenger or genomic RNA, the PCR product (DNA) of the expected size was easily visible on agarose gels stained with ethidium bromide. The specificity of the amplified DNA was confirmed by Southern and dot-blot analysis using the biotinylated oligonucleotide probe. The PCR was able to amplify the N gene sequence of purified genomic RNA from isolates of IHNV representing 5 different electropherotypes. Using the IHNV primer set, no PCR product was obtained from viral hemorrhagic septicemia virus RNA, but 2 higher molecular weight products were synthesized from hirame rhabdovirus RNA that did not hybridize with the biotinylated probe. The PCR could be efficiently performed with all IHNV genomic RNA template concentrations tested (1 ng to 1 pg). The lowest level of sensitivity was not determined. The PCR was used to amplify RNA extracted from infected cell cultures and selected tissues of Infected rainbow trout. The combination of PCR and nucleic acid probe promises to provide a detection method for IHNV that is rapid, h~ghly specific, and sensitive.

Diseases of Aquatic Organisms

Preliminary characterization of a virus causing infectious anemia among stocks of salmonid fish in the western United States

Since 1982, anemias occurring in stocks of yearling coho ( Oncorhynchus kisutch ) and chinook salmon (Oncorhynchus tshawytscha ) have been associated with serious losses at hatcheries in the Pacific Northwest, USA. The anemia is often accompanied by infections with external fungus (e.g. Saprolegnia) or the bacterial pathogens Cytophaga psychrophila or Renibacterium salmoninarum (Holt and Rohovec 1984, Leek 1987). The losses associated with the anemia are thought to be caused by these secondary infections. Blood smears that were made from anemic fish and stained with Giemsa or pinacyanol chloride showed erythrocytic inclusions ranging in size from 1 to 8 um and varying in number per cell. Thin sections of infected red blood cells (RBC) examined by electron microscopy revealed virus particles approximately 70 nm in diameter. The virions were scattered in the cytoplasm of the RBC or contained within membrane bound organelles. These virus particles were morphologically distinct from the iridovirus, erythrocytic necrosis virus (ENV), which is also associated with anemia (Holt and Rohovec 1984, Leek 1987). Evidence suggests that the etiological agent of this new anemic disease, termed erythrocytic inclusion body syndrome (EIBS) by Leek (1987), is a previously undescribed virus infecting salmon. The purpose of this study was to experimentally transmit the disease to healthy fish, to determine the blood parameters associated with infection, and to investigate the nature of the virus associated with EIBS.

Book chapter

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