Analysis of Renibacterium salmoninarum antigen production in situ.

Turaga, Prasad S. D.; Wiens, Gregory D.; Kaattari, Stephen L.

doi:10.3147/jsfp.22.209

Cited by 45 publications

(47 citation statements)

References 16 publications

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“…The p57 antigen remains circulating in the serum of the fish after infection, and in vitro assays have shown that p57 suppresses the humoral response in fish (Turaga et al 1987). When fish were immunised with formalin-killed cells that had been stripped of p57 by incubation at 37"C, the antibody titres were 20-fold higher than those of fish immunised with formalinkilled cells with p57 (Wood & Kaattari 1996).…”

Section: Introductionmentioning

confidence: 99%

Production of the 57 kDa major surface antigen by a non-agglutinating strain of the fish pathogen Renibacterium salmoninarum

Senson¹,

Stevenson²

1999

Dis. Aquat. Org.

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ABSTRACT:The major surface antigen of Renibacterium salmoninarum, p57, is associated with cell autoagglutination and imphcated as a virulence factor in fish infections. An autoagglutinating strain, JD24, caused 92% mortality when 2 X lo7 cells were injected intraperitoneally into rainbow trout Oncorhynchus mykiss, while a non-agglutinating strain, MT 239, produced only 7 % mortality after 100 d. The p57 antigen was present in the supernates of broth cultures of both strains when examined by western immunoblotting, and the gene for p57 was detected in both strains by PCR. Electron microscopy of cryopreserved thin sections showed an amorphous layer associated with the cell surface of JD24 which was not seen with MT 239. While p57 from JD24 could reassociate with cells of both strains, p57 from MT 239 failed to restore haemagglutination activity to either strain. Biotinylation of bacterial surfaces demonstrated the presence of a carbohydrate component of p57 from JD24 which was absent from the p57 produced by MT 239. The higher virulence of JD24 may depend not only on the production of p57, but also its direct association with the bacterial cell surface.

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Section: Introductionmentioning

confidence: 99%

Production of the 57 kDa major surface antigen by a non-agglutinating strain of the fish pathogen Renibacterium salmoninarum

Senson¹,

Stevenson²

1999

Dis. Aquat. Org.

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“…Because several weeks are required to culture R. salmoninarum, 3 samples are tested for R. salmoninarum cells by the flourescent antibody test (FAT) 34 or for soluble antigens of the bacterium by an enzyme-linked immunosorbent assay (ELISA). 35,41 To detect and quantify R. salmoninarum in ovarian fluid, a modified FAT was developed in which the bacteria are concentrated on a membrane filter prior to staining (MF-FAT). 11 Recently, the polymerase chain reaction (PCR) technique has shown promise for detecting very low levels of R. salmoninarum, 6,24,27,30 but it lacks the quantitative aspect of the MF-FAT and the ELISA.…”

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confidence: 99%

Comparison of the Membrane-Filtration Fluorescent Antibody Test, the Enzyme-Linked Immunosorbent Assay, and the Polymerase Chain Reaction to Detect Renibacterium Salmoninarum in Salmonid Ovarian Fluid

1998

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Abstract. Ovarian fluid samples from naturally infected chinook salmon (Oncorhynchus tshawytscha) were examined for the presence of Renibacterium salmoninarum by the membrane-filtration fluorescent antibody test (MF-FAT), an antigen capture enzyme-linked immunosorbent assay (ELISA), and a nested polymerase chain reaction (PCR). On the basis of the MF-FAT, 64% (66/103) samples contained detectable levels of R. salmoninarum cells. Among the positive fish, the R. salmoninarum concentrations ranged from 25 cells/ml to 4.3 ϫ 10 9 cells/ml. A soluble antigenic fraction of R. salmoninarum was detected in 39% of the fish (40/103) by the ELISA. The ELISA is considered one of the most sensitive detection methods for bacterial kidney disease in tissues, yet it did not detect R. salmoninarum antigen consistently at bacterial cell concentrations below about 1.3 ϫ 10 4 cells/ml according to the MF-FAT counts. When total DNA was extracted and tested in a nested PCR designed to amplify a 320-base-pair region of the gene encoding a soluble 57-kD protein of R. salmoninarum, 100% of the 100 samples tested were positive. The results provided strong evidence that R. salmoninarum may be present in ovarian fluids thought to be free of the bacterium on the basis of standard diagnostic methods.Bacterial kidney disease (BKD), caused by the gram-positive bacterium Renibacterium salmoninarum, affects wild and cultured salmonid stocks worldwide. 13,17,21 The bacterium is unique because not only can it be transmitted horizontally 31 like many other fish pathogenic microorganisms, 40 but also it can be transmitted vertically in association with eggs from infected parents. 8,15 Several mechanisms have been postulated for the egg-associated transmission of R. salmoninarum. 7,23 Many consider vertical transmission to be a consequence of maternal R. salmoninarum infections 16,34 because there is no experimental evidence to date for a contribution by an infected male. 16 As salmonid females reach sexual maturity, R. salmoninarum infections may be found in several organs and body fluids, 33 including the ovarian fluid that surrounds maturing eggs. 22,34 The relative importance of infections in various organs to the success of vertical transmission is poorly understood, but the ovarian fluid can contain more than 1 ϫ 10 9 R. salmoninarum cells/ml, 34 suggesting that this fluid may play a key role in eggassociated transmission of BKD. Whereas the prevalence of R. salmoninarum infections among fish in different progeny groups generally increases as the concentrations of the bacterium in the ovarian fluid be- Received for publication July 14, 1997. come higher, 22,34 evidence exists that vertical transmission of this bacterium may also occur when there are very low numbers of bacterial cells in the ovarian fluid. Renibacterium salmoninarum infections have been detected in chinook salmon (Oncorhynchus tshawytscha) smolts that were the progeny of females with as few as 28 bacterial cells/ml in their ovarian fluid. 22 Broodstock segregation has shown prom...

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“…salmoninarum. Similarly, a study by Turaga et al (1987) showed that the most prominent antigen found during the course of an R. salmoninarum infection had a molecular mass of approximately 60 kDa.…”

Section: Discussionmentioning

confidence: 85%

Purification, and biochemical and structural characterization of a fimbrial haemagglutinin of Renibacterium salmoninarum

Dubreuil¹,

Jacques²,

Graham

et al. 1990

Journal of General Microbiology

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~~ ~ ~~~ ~Renibacterium salmoninarum was shown to possess peritrichous fimbriae. Electron microscopy of strains FMV 84-01 and ATCC 332WT revealed short, flexible fimbriae less than 2 nm in diameter. These surface appendages were isolated from the bacteria by a procedure involving water extraction and urea solubilization. The fimbrin was purified to homogeneity by Fast Pressure Liquid Chromatography, and shown by SDS-PAGE to be a protein of 57 kDa. Isoelectric focusing under non-denaturing conditions indicated a PI of 4.8. The protein had an amino acid composition rich in glycine, Asx (aspartic acid and asparagine), valine and alanine; methionine was absent.Approximately 33 % of the amino acid residues were hydrophobic. Immunoblotting using a polyclonal antiserum raised against whole cells showed that the 57 kDa protein was the immunodominant antigen on the cell surface.Immunogold labelling using polyclonal antibodies raised against the fimbrin revealed an alignment of gold particles along the fimbriae. Purified fimbriae caused agglutination of rabbit erythrocytes and antifimbrial serum inhibited this haemagglutination. Altogether the results indicate that the fimbriae on the surface of R. salmoninarum are responsible for the haemagglutinating activity.

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Analysis of Renibacterium salmoninarum antigen production in situ.

Cited by 45 publications

References 16 publications

Production of the 57 kDa major surface antigen by a non-agglutinating strain of the fish pathogen Renibacterium salmoninarum

Production of the 57 kDa major surface antigen by a non-agglutinating strain of the fish pathogen Renibacterium salmoninarum

Comparison of the Membrane-Filtration Fluorescent Antibody Test, the Enzyme-Linked Immunosorbent Assay, and the Polymerase Chain Reaction to Detect Renibacterium Salmoninarum in Salmonid Ovarian Fluid

Purification, and biochemical and structural characterization of a fimbrial haemagglutinin of Renibacterium salmoninarum

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