DAO 97:197-205 (2012)  -  DOI: https://doi.org/10.3354/dao02403

Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis 

A. J. Yañez1,*, K. Valenzuela1, H. Silva1, J. Retamales2, A. Romero3, R. Enriquez3, J. Figueroa1, A. Claude1, J. Gonzalez4, R. Avendaño-Herrera2, J. G. Carcamo

1Laboratorio de Enzimología, Instituto de Bioquímica, and 3Instituto de Patología, Facultad de Ciencias, Universidad Austral de Chile, Valdivia, Chile
2Laboratorio de Patología de Organismos Acuáticos y Biotecnología Acuícola, Facultad de Ciencias Biológicas, Universidad Andrés Bello, Viña del Mar, Chile
4EWOS Chile Alimentos Ltda, Coronel, Chile

ABSTRACT: Piscirickettsiosis or salmonid rickettsial septicaemia (SRS) caused by Piscirickettsia salmonis constitutes one of the main problems in farmed salmonid and marine fishes. Since the first reports of the disease, it has been successfully isolated and maintained in eukaryotic cell-­culture systems, but these systems are time-consuming, the media are costly, and eliminating heavily contaminated host cell debris is difficult. In this report, we describe a marine-based broth supplemented with L-cysteine, named AUSTRAL-SRS broth, that facilitates superior growth of P. salmonis strains. Strains reached an optical density of approximately 1.8 when absorbance was measured at 600 nm after 6 d incubation at 18°C. Several passages (n = 6) did not alter the culture kinetics. We report for the first time the purification of DNA, lipopolysaccharide (LPS) and whole membrane protein obtained from P. salmonis grown in this liquid medium, and thus provide a suitable platform to simplify the preparation of P. salmonis cells for genetic and serological studies. Moreover, the results of the cytopathic effect test showed that P. salmonis grown in AUSTRAL-SRS broth maintained their virulence properties, inducing apoptosis after 3 d. This makes the medium a good candidate for the successful growth of P. salmonis and an excellent basis for the development of low cost vaccines.


KEY WORDS: Piscirickettsia salmonis · Broth · Culture


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Cite this article as: Yañez AJ, Valenzuela K, Silva H, Retamales J and others (2012) Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis . Dis Aquat Org 97:197-205. https://doi.org/10.3354/dao02403

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