Comparative Sensitivities of Diagnostic Procedures Used to Detect Bacterial Kidney Disease in Salmonid Fishes

Cipriano, Rocco C.; Starliper, Clifford E.; Schachte, John H.

doi:10.7589/0090-3558-21.2.144

Cited by 29 publications

(20 citation statements)

References 5 publications

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“…In agreement with Evelyn et al (1981), Cipriano et al (1985), Benediktsdottir et al ( 1 991) and Gudmundsdottir et al (1991), our findings showed that culture can be used to demonstrate bacteria in fish with no external or internal signs of disease, i.e. the so-called latent carriers.…”

Section: Discussionsupporting

confidence: 77%

“…Several studies have also shown culture to be a very sensitive method for detecting R.s. but other investigations have shown certain serological methods to be superior to culture with regard to sensitivity (Cipriano et al 1985, Pascho et al 1987, Sakai et al 1987, Gudmundsdottir et al 1990). Our results showed that culture can fail to confirm a suspicion of BKD based on internal findings, especially in fish that are externally 'healthy looking'.…”

Section: Discussionmentioning

confidence: 99%

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Practical aspects of bacterial culture for the diagnosis of bacterial kidney disease (BKD)

Olsen¹,

Hopp²,

Binde³

et al. 1992

Dis. Aquat. Org.

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A study was made of practical aspects of culture of Renibacterium salrnonjnarum (R.s.) from the kidney of Atlantic salmon Salmo salar. Samples were taken from fish in sea farms affected by outbreaks of bacterial kidney disease (BKD). Only 67% of the samples positive for growth of R.s. on kidney disease medium (KDM2) were positive on selective kidney disease medium (SKDM). Of the samples positive on KDM2, 98 % were identified as positive after 8 wk of incubation a t 15 "C, whereas only 82 "is of the samples positive on SKDlvl were positive at this time Growth of R.s on KDM2 produced more colonies than growth on SKDM No significant differences in isolation frequency were seen betureen culture from anterior and poster~or k~d n e y .If 3 or 4 ~n o c u l a t~o n s were made from each fish, the number of R.s.-poslt~ve f~s h increased by 44 0/,) in the lightly infected population of thls study. Both storage of the sample material on swabs in 2 different niodifications of Stuart's transport medium for 24 h, and freezing a t -20 "C for 3 to 4 mo, reduced the sensitivity of culture from lightly infected samples.

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Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 99%

Practical aspects of bacterial culture for the diagnosis of bacterial kidney disease (BKD)

Olsen¹,

Hopp²,

Binde³

et al. 1992

Dis. Aquat. Org.

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“…Although studies comparing R. salmoninarum detection methods continue to be reported as new diagnostic tests are introduced, 6,10,19,28,38,42 very few have included ovarian fluid as the test sample. 12,25,30,33 This is the first comparative study to provide strong evidence that R. salmoninarum may be present in ovarian fluids thought to be free of the bacterium on the basis of 2 standard diagnostic methods, the MF-FAT and the ELISA.…”

Section: Discussionmentioning

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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“…Consequently, many workers have preferred to diagnose BKD directly by the use of various techniques, predominantly serological, on fish extracts. Serological techniques have included the direct or indirect fluorescent antibody test (FAT) (BULLOCK and STUCKEY 1975;BULLOCK et al 1980), immunodiffusion (ID) (CHEN et al 1974;KIMURA et al 1978), staphylococcal coagglutination (CoA) (KIMURA 1978), and counterimmunoelectrophoresis (CIE) (CIPRIANO et al 1985). However, in a comparative study, isolation of R. salmoninarum in culture was found to be more sensitive than FAT or ID (EVELYN 1978) and although CIPRIANO et al (1985) reported that isolation in culture was less sensitivie than CIE or CoA, those authors stated that primary isolation followed by serological identification was the most reliable method for the detection of R .…”

Section: Introductionmentioning

confidence: 99%

Detection of Renibacterium salmoninarum by the enzyme-linked immunosorbent assay (ELISA)

Dixon

1987

J Appl Ichthyol

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An enzyme-linked immunosorbent assay (ELISA) was developed for the detection and identification of Renibacterium salmoninarum. The immune y-globulin used in the assay was absorbed with two species of cross-reacting bacteria to make a specific test system. R. salmoninarum could be detected in clin~call diseased fish within 30 minutes of preparing a kidney sam le, and thus because of its ease of use, ti; ELISA could be employed as a rapid field test for bacterial Ldney disease (BKD), although isolation of R. salmoninarum was more sensitive than the ELISA for detecting individual carrier fish. Zusammenfassung Identifizierung von Renibacterium salmoninarum mit Hive des ELISA-TestsZur Identifizierung des Renibacterium salmoninarum, dem Erreger der bakteriellen Nierenkrankheit (BKD), wurde ein ELISA-Test entwickelt, der sich als Schnelltest unter Feldbedingungen bewahrt hat. Das in diesem Test verwendete Immun y-globulin ist an kreuzreagierenden Bakterien adsorbiert und mit alkalischer Phosphatase konjugiert worden. Mit diesem Reagenz gelang es bei klinisch an der BKD erkrankten Fischen Renibacterium salmoninarum in der Niere innerhalb von 30 Minuten nachzuweisen. Bei Carriern erwies sich dagegen die Erregeranzucht als aussagekraftiger. Resume Detection de Renibacterium salmoninarum i l'aide du test ELISAUn test (ELISA) a it6 develo p i our dktecter et identifier le Renibacterium salmoninarum. L'immunoglobuline y utilisie Bans test a CtC absorbee sur deux espkces de bactiries contreriagissantes dans le but de creer un systkme de test s Ccifique. Renibacterium salmoninarum a pu ktre dkcouvert en I'espace de 30 minutes dans un khantieon pris sur le rein chez des poissons portant des sym tomes cliniques et, en raison de sa facilitk d'emploi, le test ELISA a pu ktre employe comme test rapiie pour la maladie bacterielle du rein (BKD), bien que I'isolation de R. salmoninarum ait 6t6 plus sensible que le test ELISA pour la detection du poisson porteur.

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Comparative Sensitivities of Diagnostic Procedures Used to Detect Bacterial Kidney Disease in Salmonid Fishes

Cited by 29 publications

References 5 publications

Practical aspects of bacterial culture for the diagnosis of bacterial kidney disease (BKD)

Practical aspects of bacterial culture for the diagnosis of bacterial kidney disease (BKD)

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

Detection of Renibacterium salmoninarum by the enzyme-linked immunosorbent assay (ELISA)

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