Disease caused by <i>Yersinia ruckeri</i> serotype O2b found in Chilean‐farmed coho salmon, <i>Oncorhynchus kisutch</i> (Walbaum, 1792)

Avendaño‐Herrera, Rubén; Tapia‐Cammas, Diana; Aedo, Alejandra; Saldivia, P; Ortega, Claudio Bravo; Irgang, Rute

doi:10.1111/jfd.12502

Cited by 6 publications

(6 citation statements)

References 31 publications

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“…The anti‐rabbit polyclonal antibody against V. ordalii Au3, used in confocal and flow cytometer assays, was described by Silva‐Rubio, Acevedo, et al (). In addition, anti‐ Vibrio anguillarum ATCC 43307 and anti‐ Yersinia ruckeri CECT 955 antibodies were prepared (Avendaño‐Herrera et al, ; Silva‐Rubio, Avendaño‐Herrera, Jaureguiberry, Toranzo, & Magariños, ) and used as controls of extracellular and intracellular bacteria, respectively. The purity of the microorganisms used as controls was confirmed by an indirect fluorescent antibody test.…”

Section: Methodsmentioning

confidence: 99%

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Avendaño‐Herrera

Arias‐Muñoz

Rojas

et al. 2019

Journal of Fish Diseases

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Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.

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Section: Methodsmentioning

confidence: 99%

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Avendaño‐Herrera

Arias‐Muñoz

Rojas

et al. 2019

Journal of Fish Diseases

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“…In recent years, the number of outbreaks in the farmed Atlantic salmon population has substantially increased. The reasons for the most recent outbreaks remain unclear, and Y. ruckeri infections are nowadays a major challenge facing the Norwegian aquaculture industry, similar to other countries such as Australia (Barnes et al, 2016), Chile (Avendaño-Herrera et al, 2017), and Scotland .…”

Section: Introductionmentioning

confidence: 99%

The repeat structure of two paralogous genes, Yersinia ruckeri invasin (yrInv) and a “Y. ruckeri invasin-like molecule”, (yrIlm) sheds light on the evolution of adhesive capacities of a fish pathogen

Wróbel

Ottoni

Leo

et al. 2018

Journal of Structural Biology

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Inverse autotransporters comprise the recently identified type Ve secretion system and are exemplified by intimin from enterohaemorrhagic Escherichia coli and invasin from enteropathogenic Yersiniae. These proteins share a common domain architecture and promote bacterial adhesion to host cells. Here, we identified and characterized two putative inverse autotransporter genes in the fish pathogen Yersinia ruckeri NVH_3758, namely yrInv (for Y. ruckeri invasin) and yrIlm (for Y. ruckeri invasin-like molecule). When trying to clone the highly repetitive genes for structural and functional studies, we experienced problems in obtaining PCR products. PCR failures and the highly repetitive nature of inverse autotransporters prompted us to sequence the genome of Y. ruckeri NVH_3758 using PacBio sequencing, which produces some of the longest average read lengths available in the industry at this moment. According to our sequencing data, YrIlm is composed of 2603 amino acids (7812bp) and has a molecular mass of 256.4kDa. Based on the new genome information, we performed PCR analysis on four non-sequenced Y. ruckeri strains as well as the sequenced. Y. ruckeri type strain. We found that the genes are variably present in the strains, and that the length of yrIlm, when present, also varies. In addition, the length of the gene product for all strains, including the type strain, was much longer than expected based on deposited sequences. The internal repeats of the yrInv gene product are highly diverged, but represent the same bacterial immunoglobulin-like domains as in yrIlm. Using qRT-PCR, we found that yrIlm and yrInv are differentially expressed under conditions relevant for pathogenesis. In addition, we compared the genomic context of both genes in the newly sequenced Y. ruckeri strain to all available PacBio-sequenced Y. ruckeri genomes, and found indications of recent events of horizontal gene transfer. Taken together, this study demonstrates and highlights the power of Single Molecule Real-Time technology for sequencing highly repetitive proteins, and sheds light on the genetic events that gave rise to these highly repetitive genes in a commercially important fish pathogen.

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“…In recent years, the number of outbreaks in the farmed Atlantic salmon population has substantially increased. The reasons for the most recent outbreaks remain unclear, and Y. ruckeri infections are nowadays a major challenge facing the Norwegian aquaculture industry, similar to other countries such as Australia (Barnes et al, 2016 ), Chile (Avendaño-Herrera et al, 2017 ), and Scotland (Ormsby et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

pYR4 From a Norwegian Isolate of Yersinia ruckeri Is a Putative Virulence Plasmid Encoding Both a Type IV Pilus and a Type IV Secretion System

Wróbel

Ottoni

Leo

et al. 2018

Front. Cell. Infect. Microbiol.

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Enteric redmouth disease caused by the pathogen Yersinia ruckeri is a significant problem for fish farming around the world. Despite its importance, only a few virulence factors of Y. ruckeri have been identified and studied in detail. Here, we report and analyze the complete DNA sequence of pYR4, a plasmid from a highly pathogenic Norwegian Y. ruckeri isolate, sequenced using PacBio SMRT technology. Like the well-known pYV plasmid of human pathogenic Yersiniae, pYR4 is a member of the IncFII family. Thirty-one percent of the pYR4 sequence is unique compared to other Y. ruckeri plasmids. The unique regions contain, among others genes, a large number of mobile genetic elements and two partitioning systems. The G+C content of pYR4 is higher than that of the Y. ruckeri NVH_3758 genome, indicating its relatively recent horizontal acquisition. pYR4, as well as the related plasmid pYR3, comprises operons that encode for type IV pili and for a conjugation system (tra). In contrast to other Yersinia plasmids, pYR4 cannot be cured at elevated temperatures. Our study highlights the power of PacBio sequencing technology for identifying mis-assembled segments of genomic sequences. Comparative analysis of pYR4 and other Y. ruckeri plasmids and genomes, which were sequenced by second and the third generation sequencing technologies, showed errors in second generation sequencing assemblies. Specifically, in the Y. ruckeri 150 and Y. ruckeri ATCC29473 genome assemblies, we mapped the entire pYR3 plasmid sequence. Placing plasmid sequences on the chromosome can result in erroneous biological conclusions. Thus, PacBio sequencing or similar long-read methods should always be preferred for de novo genome sequencing. As the tra operons of pYR3, although misplaced on the chromosome during the genome assembly process, were demonstrated to have an effect on virulence, and type IV pili are virulence factors in many bacteria, we suggest that pYR4 directly contributes to Y. ruckeri virulence.

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Disease caused by Yersinia ruckeri serotype O2b found in Chilean‐farmed coho salmon, Oncorhynchus kisutch (Walbaum, 1792)

Cited by 6 publications

References 31 publications

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

The repeat structure of two paralogous genes, Yersinia ruckeri invasin (yrInv) and a “Y. ruckeri invasin-like molecule”, (yrIlm) sheds light on the evolution of adhesive capacities of a fish pathogen

pYR4 From a Norwegian Isolate of Yersinia ruckeri Is a Putative Virulence Plasmid Encoding Both a Type IV Pilus and a Type IV Secretion System

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