Immunological characterization of a bacterial protein isolated from salmonid fish naturally infected with Piscirickettsia salmonis

Marshall, Sergio H.; Conejeros, Pablo; Zahr, M.; Olivares, Jorge; Gómez, Fernando; Cataldo, Patricio; Henríquez, Vitalia

doi:10.1016/j.vaccine.2006.11.035

Cited by 40 publications

(35 citation statements)

References 34 publications

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“…The prototype strain LF-89 has been successfully grown and cloned as single colonies on Sheep Blood Agar plates (BFCG) as well as on Fish Blood Broth media (BB). The identity of the colonies was determined by PCR against the ITS region of the ribosomal DNA and also against the coding sequence of the ChaP.s functional protein (Marshall et al 2007). These results were further confirmed by western blot analysis protein profiles using a wide array of anti-P. salmonis antibodies and also by direct and indirect immunofluorescence analysis.…”

Section: Introductionmentioning

confidence: 67%

“…Piscirickettsia salmonis is the etiological agent of the Salmonid Rickettsial Septicaemia (SRS) or Piscirickettsiosis, which is an aggressive infectious disease affecting salmonid fish since the late '80s (Bravo and Campos 1989;Graggero et al 1995, Marshall et al 2007. Additionally, non-salmonids have been affected by P. salmonis or P. salmonis-like organisms.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, the presence of this bacteria has been detected in specimens of white Seabass (Atractoscion nobilis) on the Southern California coast (Arkush KD et al 2005). European Seabass (Dicentrarchus labrax) in Greece have been affected by a pathogen similar to P. salmonis (Athanassopoulou et al 2004); also in Hawaii, Tilapia populations (oreochromis mossambicus and Sarotherodon melanotheron), both free-living as well as reared fish have suffered a Piscirickettsiosis-type disease (Mauel et al 2003), which suggests that the expansion of this agent to other fish species of commercial importance has already begun (Marshall et al 2007).…”

Section: Introductionmentioning

confidence: 99%

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Additional evidence of the facultative intracellular nature of the fish bacterial pathogen Piscirickettsia salmonis

2009

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RESuMENPiscirickettsia salmonis es un microorganismo altamente contagioso y virulento que afecta a la salmonicultura mundial desde el último tercio del siglo pasado y del cual sus mecanismos de sobrevivencia intracelular son completamente desconocidos. Después de algunos reportes recientes en donde se cuestiona su condición de intracelular obligado, hemos decidido mostrar evidencia adicional para cambiar este paradigma, llevando a cabo experimentos tanto clásicos como moleculares que confirman su naturaleza de intracelular facultativo. En este reporte se demuestra inequívocamente que la bacteria recuperada desde cultivos celulares infectados, de placas de agar o de medio líquido, es el mismo organismo, el cual cumple con los postulados de Koch. Además, análisis genéticos y proteómicos confirman que la bacteria obtenida de diferentes fuentes de crecimiento corresponde a la misma cepa tipo LF-89, la que fue originalmente descrita por Fryer en 1992. Sin embargo, el crecimiento de la bacteria, tanto en medios libres de células como en cultivo celular, es subóptimo, por lo cual se requieren más análisis para entender completamente la biología del patógeno. Interesantemente, en este trabajo se logró la mantención de la bacteria en medio líquido, pero a una tasa muy baja de crecimiento. En conclusión, y sumado a reportes anteriores, hemos confirmado la naturaleza intracelular facultativa de este patógeno de peces de cultivo.Palabras clave: Piscirickettsia salmonis, intracelular facultativo, genómica.

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Section: Introductionmentioning

confidence: 67%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Additional evidence of the facultative intracellular nature of the fish bacterial pathogen Piscirickettsia salmonis

2009

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“…An anti-ChaPs (HSP60 of P. salmonis) antibody was prepared in the laboratory and was used as a control to recognize the bacterium (Marshall et al 2007). Detection was performed under the same conditions as described above, although an antimouse Alexa Fluor 647-conjugated antibody was used for specific detection.…”

Section: Confocal Laser Scanning Microscopy (Clsm) Analysis Of Flagelmentioning

confidence: 99%

“…5). As a control for the identification of P. salmonis in CLSM observations, an anti-ChaPs antibody (HSP60 of P. salmonis) was used, as ChaPs is an abundant protein in P. salmonis (Marshall et al 2007). This approach was further supported with TEM analysis, where FlaA was only found located on the periphery and in the cytosol of the host cell-released bacteria (Fig.…”

Section: Cellular Localization Of the Flagellin Monomermentioning

confidence: 99%

Expression of flagellin and key regulatory flagellar genes in the non-motile bacterium Piscirickettsia salmonis

Carril

Gómez²,

Marshall³

2017

Dis. Aquat. Org.

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The Piscirickettsia salmonis genome was screened to evaluate potential flagellarelated open reading frames, as well as their genomic organization and eventual expression. A complete and organized set of flagellar genes was found for P. salmonis, although no structural flagellum has ever been reported for this bacterium. To gain further understanding, the hierarchical flagellar cascade described for Legionella pneumophila was used as a reference model for putative analysis in P. salmonis. Specifically, 5 of the most relevant genes from this cascade were chosen, including 3 regulatory genes (fleQ, triggers the cascade; fliA, regulates the σ28-coding gene; and rpoN, an RNA polymerase-dependent gene) and 2 terminal structural genes (flaA and flaB, flagellin and a flagellin-like protein, respectively). Kinetic experiments evaluated gene expressions over time, with P. salmonis assessed in 2 liquid, cell-free media and during infection of the SHK-1 fish cell line. Under all conditions, the 5 target genes were primarily expressed during early growth/infection and were differentially expressed when bacteria encountered environmental stress (i.e. a high-salt concentration). Intriguingly, the flagellin monomer was fully expressed under all growth conditions and was located near the bacterial membrane. While no structural flagellum was detected under any condition, the recombinant flagellin monomer induced a proinflammatory response in SHK-1 cells, suggesting a possible immunomodulatory function. The potential implications of these observations are discussed in the context of P. salmonis biology and pathogenic potential.

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Vaccination against Piscirickettsiosis

Marshall

Tobar²

2014

Fish Vaccination

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Immunological characterization of a bacterial protein isolated from salmonid fish naturally infected with Piscirickettsia salmonis

Cited by 40 publications

References 34 publications

Additional evidence of the facultative intracellular nature of the fish bacterial pathogen Piscirickettsia salmonis

Additional evidence of the facultative intracellular nature of the fish bacterial pathogen Piscirickettsia salmonis

Expression of flagellin and key regulatory flagellar genes in the non-motile bacterium Piscirickettsia salmonis

Vaccination against Piscirickettsiosis

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