ELISA-Based Segregation of Adult Spring Chinook Salmon for Control of Bacterial Kidney Disease, Annual Report FY 1989.

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Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum, is a serious disease of salmonid fish worldwide. The disease has a major impact on spring chinook salmon populations in the Columbia River system. There is strong evidence that R. safmoninarum can be transmitted from parent to progeny, and therefore culling of gametes from infected parents should obviate this mode of transmission. This report presents the results from the first year of our four year study to investigate segregation of broodstock as a tool for controlling BKD. The segregations will use Enzyme-Linked Immunosorbent Assays (ELISAs) as detection systems to identify, in tissues ... continued below

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48 pages

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Kaattari, Stephen L. & Winton, James R. December 1, 1989.

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Description

Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum, is a serious disease of salmonid fish worldwide. The disease has a major impact on spring chinook salmon populations in the Columbia River system. There is strong evidence that R. safmoninarum can be transmitted from parent to progeny, and therefore culling of gametes from infected parents should obviate this mode of transmission. This report presents the results from the first year of our four year study to investigate segregation of broodstock as a tool for controlling BKD. The segregations will use Enzyme-Linked Immunosorbent Assays (ELISAs) as detection systems to identify, in tissues of infected fish, proteins produced by R. salmoninarum. A first step in the development of the described detection systems was the optimization of the production of important antigenic proteins from R. salmoninarum. Different culture media were qualitatively and quantitatively evaluated for their ability to support production of cellular and soluble proteins. The major factor affecting antigen quality was the presence and absence of calf serum. Media components and R. salmoninarum growth products could not be separated during harvest of proteins from the cultures containing serum. This caused problems with the quantitation of actual bacterial proteins in the preparation. Thus media without serum is currently employed. Two independent ELISA techniques for the identification of infected parents were examined. One technique is based on polyclonal antisera produced in rabbits and the second is based on mouse monoclonal antibodies (Mabs). To develop the latter system, several Mabs against a major R. salmoninarum antigenic protein were produced. These Mabs were used for the detection of R. salmoninarum antigens in infected fish and also to characterize proteins produced by the bacterium. Both ELISAs were deemed suitable for the segregation of parents into the high and low BKD groups required for this study. An alternate system for the detection of R. salmoninarum proteins, the Western blot, was also investigated, This technique was 100 to 1000 fold less sensitive than either ELISA system and therefore will be useful only for confirmation of highly positive tissues. Future work will attempt to increase the sensitivity of the Western blotting system. Finally, two hatcheries were identified for use in the described segregation experiments. The Carson National Fish Hatchery (Skamania County, WA) and the Marion Forks Fish Hatchery (Linn County, OR) will be used for the experiments.

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48 pages

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  • Report No.: DOE/BP-95906-1
  • Grant Number: DE-FG79-95905
  • DOI: 10.2172/920196 | External Link
  • Office of Scientific & Technical Information Report Number: 920196
  • Archival Resource Key: ark:/67531/metadc900820

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  • December 1, 1989

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  • Sept. 27, 2016, 1:39 a.m.

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Kaattari, Stephen L. & Winton, James R. ELISA-Based Segregation of Adult Spring Chinook Salmon for Control of Bacterial Kidney Disease, Annual Report FY 1989., report, December 1, 1989; United States. (digital.library.unt.edu/ark:/67531/metadc900820/: accessed August 17, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.