Evaluation and development of diagnostic methods for Renibacterium salmoninarum causing bacterial kidney disease (BKD) in the UK

被引:44
作者
Bruno, David
Collet, Bertrand
Turnbull, Anna
Kilburn, Rachel
Walker, Amanda
Pendrey, Daniel
McIntosh, Alison
Urquhart, Katy
Taylor, Gillian
机构
[1] FRS, Marine Lab, Aberdeen AB11 9DB, Scotland
[2] Cefas Weymouth Lab, Weymouth DT4 8UB, England
关键词
Renibacterium salmoninarum; diagnosis; culture; ELISA; non-destructive real-time qPCR; IFAT;
D O I
10.1016/j.aquaculture.2007.04.057
中图分类号
S9 [水产、渔业];
学科分类号
0908 ;
摘要
Government agencies responsible for statutory fish health programmes in the UK (FRS Marine Laboratory, Aberdeen and Cefas, Weymouth) participated in a comparative study for the detection of Renibacterium salmoninarum or antigen using culture, Gram stained imprints and ELISA from fanned Atlantic salmon, Salmo salar and rainbow trout, Oncorhynchus mykiss. FRS conducted additional tests, namely H&E and Gram staining of tissue sections, indirect fluorescent antibody (IFAT) and a quantitative real-time PCR (qPCR) assay to detect Msa2 genes of R. salmoninarum (RS) and the Elongation factor alpha I (ELF) gene of salmonids from genomic DNA. Three Scottish farms were selected for the study based on current or past infection. ANOVA (analysis of variance) was used to analyse data to determine comparative sensitivity of certain paired tests. Results show good correlation by independent ('blind') testing from each laboratory. Isolation using Mueller Hinton with added cysteine (MHCA) resulted in both laboratories identifying the same fish as culture positive; using this medium FRS reported one additional fish as positive. Isolation using selective kidney disease medium (SKDM) demonstrated R. salmoninarum in the same fish as MHCA. Isolation of R. salmoninarum by culture was supported by other tests. Both laboratories identified the same fish as BKD positive from Gram stained imprints. Cefas identified one additional fish by this method. ELISA testing by both laboratories identified 5/27 fish from farm 1. Additional fish were reported positive at farm 2 by FRS. A ratio RS/ELF of < 0.000013 represented the value where other tests used in this exercise were negative. A ratio of > 0.000056 generally correlated with other tests used in this exercise. Two fish identified as ELISA positive from farm 2 were not supported by other tests including qPCR on blood samples; however re-testing using kidney from these fish gave a positive PCR result. The qPCR results showed 40.7, 22.2 and 1.3% prevalence from sites 1, 2 and 3 respectively. Significant or highly significant values were obtained for ANOVA analysis using qPCR v ELISA or H&E staining for farms I and 2. Positive IFAT staining of kidney, liver and spleen was demonstrated for fish at farm I and these results correlated with other methods. A good correlation between agencies is reported as well as correlation between methods, however, it is recognised that further evaluation and testing is necessary. (C) 2007 Published by Elsevier B.V.
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收藏
页码:114 / 122
页数:9
相关论文
共 32 条
[1]  
AUSTIN B, 1983, FEMS MICROBIOL LETT, V17, P111, DOI 10.1111/j.1574-6968.1983.tb00383.x
[2]   INCUBATION-TIME FOR THE CULTIVATION OF RENIBACTERIUM-SALMONINARUM FROM ATLANTIC SALMON, SALMO-SALAR L, BROODFISH [J].
BENEDIKTSDOTTIR, E ;
HELGASON, S ;
GUDMUNDSDOTTIR, S .
JOURNAL OF FISH DISEASES, 1991, 14 (01) :97-102
[3]   USE OF THE POLYMERASE CHAIN-REACTION (PCR) TO DETECT DNA FROM RENIBACTERIUM-SALMONINARUM WITHIN INDIVIDUAL SALMONID EGGS [J].
BROWN, LL ;
IWAMA, GK ;
EVELYN, TPT ;
NELSON, WS ;
LEVINE, RP .
DISEASES OF AQUATIC ORGANISMS, 1994, 18 (03) :165-171
[4]  
BRUNO D W, 1986, Aquaculture and Fisheries Management, V17, P185, DOI 10.1111/j.1365-2109.1986.tb00100.x
[5]   The occurrence of Renibacterium salmoninarum within vaccine adhesion components from Atlantic salmon, Salmo salar L. and coho salmon, Oncorhynchus kisutch Walbaum [J].
Bruno, DW ;
Brown, LL .
AQUACULTURE, 1999, 170 (01) :1-5
[6]   Prevalence and diagnosis of bacterial kidney disease (BKD) in Scotland between 1990 and 2002 [J].
Bruno, DW .
DISEASES OF AQUATIC ORGANISMS, 2004, 59 (02) :125-130
[8]   Renibacterium salmoninarum bar forms:: characterization, occurrence, and evidence of a host response to a R-salmoninarum infection [J].
Cvitanich, JD .
JOURNAL OF FISH DISEASES, 2004, 27 (04) :193-211
[9]  
Evelyn T. P. T., 1977, Bulletin de l'Office International des Epizooties, V87, P511
[10]   A SIMPLE TECHNIQUE FOR ACCELERATING THE GROWTH OF THE KIDNEY-DISEASE BACTERIUM RENIBACTERIUM-SALMONINARUM ON A COMMONLY USED CULTURE-MEDIUM (KDM2) [J].
EVELYN, TPT ;
BELL, GR ;
PROSPERIPORTA, L ;
KETCHESON, JE .
DISEASES OF AQUATIC ORGANISMS, 1989, 7 (03) :231-234