Evaluation of four polymerase chain reaction primer pairs for the detection of Edwardsiella tarda in turbot

Castro, Nuria; Toranzo, Alicia E.; Núñez, Soledad; Osorio, Carlos R.; Magariños, Beatríz

doi:10.3354/dao02203

Cited by 28 publications

(26 citation statements)

References 8 publications

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“…According to an earlier study, the minimum density of E . tarda that can be detected in a mixed bacterial culture is 10 5 CFU/mL (Castro et al ), which is similar to the efficiency of our multiplex PCR assay showing that the minimum detection limits for E . tarda and V .…”

Section: Discussionsupporting

confidence: 87%

Identification and Detection of the Pathogenic Bacteria Responsible for Swollen Abdomen Disease in Cultured Turbot, Scophthalmus maximus, and Flounder, Paralichthys olivaceus

Xue

Sun

2017

J World Aquaculture Soc

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Swollen abdomen disease (SAD) seriously threatens the aquaculture of turbots and flounders. Two dominant bacterial strains, FS1 and FS2, were isolated from the livers and kidneys of fish with diagnosed SAD. Applications of biochemical analyses, sequence analyses of 16S ribosomal RNA gene and heat shock protein 60 gene revealed two distinct pathogenic bacterial species, Edwardsiella tarda and Vibrio alginolyticus. These two hypothesized SAD‐causing pathogens were validated by challenge trials on flounder, Paralichthys olivaceus. Challenges with E. tarda and V. alginolyticus demonstrated lethal dose 50 (LD50) values at 1.51 × 105 colony‐forming units (CFU) and 1.05 × 105 CFU, respectively. To develop a rapid SAD diagnosis method in flounders and turbots, a multiplex polymerase chain reaction (PCR) assay method was developed to simultaneously detect E. tarda and V. alginolyticus. Our multiplex PCR assay successfully detected as low as 105 CFU/mL E. tarda and V. alginolyticus in flounders and turbots. No other common fish pathogens were detected with the multiplex PCR, suggesting a high specificity of this assay. The multiplex PCR assay developed in this study showed a great reliability in detecting SAD‐causing bacterial pathogens. Further optimization of this assay may contribute to the development of a novel SAD diagnosis tool in aquaculture.

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Section: Discussionsupporting

confidence: 87%

Identification and Detection of the Pathogenic Bacteria Responsible for Swollen Abdomen Disease in Cultured Turbot, Scophthalmus maximus, and Flounder, Paralichthys olivaceus

Xue

Sun

2017

J World Aquaculture Soc

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“…) as well as by PCR‐specific analysis (Castro et al . ). E. tarda strains were routinely grown on TSA (trypticase soy agar, Difco) at 25 °C.…”

Section: Methodsmentioning

confidence: 97%

Insights into the virulence‐related genes of Edwardsiella tarda isolated from turbot in Europe: genetic homogeneity and evidence for vibrioferrin production

Castro

Osorio

Buján

et al. 2015

Journal of Fish Diseases

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Edwardsiella tarda has long been known as a pathogen that causes severe economic losses in aquaculture industry. Insights gained on E. tarda pathogenesis may prove useful in the development of new methods for the treatment of infections as well as preventive measures against future outbreaks. In this report, we have established the correlation between the presence of virulence genes, related with three aspects typically involved in bacterial pathogenesis (chondroitinase activity, quorum sensing and siderophore-mediated ferric uptake systems), in the genome of E. tarda strains isolated from turbot in Europe and their phenotypic traits. A total of 8 genes were tested by PCR for their presence in 73 E. tarda isolates. High homogeneity was observed in the presence/absence pattern of all the strains. Positive results in the amplification of virulence-related genes were correlated with the detection of chondroitinase activity in agar plates, in vivo AHL production during fish infection and determination of type of siderophore produced by E. tarda. To the best of our knowledge, this is the first study carried out with European strains on potential virulence factors. Furthermore, we demonstrated for the first time that E. tarda produces the siderophore vibrioferrin.

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“…Chang et al [54] developed a multiplex nested PCR for four important fish pathogens in subtropical Asia that can simultaneously detect A. hydrophila , E. tarda , Photobacterium damselae and Streptococcus iniae from pure colonies and tissue homogenates. In addition, a primer set, evaluated using 53 E. tarda strains isolated from various sources and 18 representative strains of related and unrelated bacterial species, was shown capable of detecting two cells from pure culture and 3 × 10 2 cells in seeded turbot tissues [55]. …”

Section: Pathology and Diagnosismentioning

confidence: 99%

Pathogenesis of and strategies for preventing Edwardsiella tarda infection in fish

Park¹,

Aoki²,

Jung³

2012

Vet Res

319

185

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Edwardsiella tarda is one of the serious fish pathogens, infecting both cultured and wild fish species. Research on edwardsiellosis has revealed that E. tarda has a broad host range and geographic distribution, and contains important virulence factors that enhance bacterial survival and pathogenesis in hosts. Although recent progress in edwardsiellosis research has enabled the development of numerous, highly effective vaccine candidates, these efforts have not been translated into a commercialized vaccine. The present review aims to provide an overview of the identification, pathology, diagnosis and virulence factors of E. tarda in fish, and describe recent strategies for developing vaccines against edwardsiellosis. The hope is that this presentation will be useful not only from the standpoint of understanding the pathogenesis of E. tarda, but also from the perspective of facilitating the development of effective vaccines.

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Evaluation of four polymerase chain reaction primer pairs for the detection of Edwardsiella tarda in turbot

Cited by 28 publications

References 8 publications

Identification and Detection of the Pathogenic Bacteria Responsible for Swollen Abdomen Disease in Cultured Turbot, Scophthalmus maximus, and Flounder, Paralichthys olivaceus

Identification and Detection of the Pathogenic Bacteria Responsible for Swollen Abdomen Disease in Cultured Turbot, Scophthalmus maximus, and Flounder, Paralichthys olivaceus

Insights into the virulence‐related genes of Edwardsiella tarda isolated from turbot in Europe: genetic homogeneity and evidence for vibrioferrin production

Pathogenesis of and strategies for preventing Edwardsiella tarda infection in fish

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