title: Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis .
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.
authors.affiliation: Laboratorio de Enzimología, Instituto de Bioquímica, Facultad de Ciencias, Universidad Austral de Chile, Valdivia, Chile. ayanez@uach.cl
authors.forename: A J
authors.initials: AJ
authors.lastname: Yañez
authors.forename: K
authors.initials: K
authors.lastname: Valenzuela
authors.forename: H
authors.initials: H
authors.lastname: Silva
authors.forename: J
authors.initials: J
authors.lastname: Retamales
authors.forename: A
authors.initials: A
authors.lastname: Romero
authors.forename: R
authors.initials: R
authors.lastname: Enriquez
authors.forename: J
authors.initials: J
authors.lastname: Figueroa
authors.forename: A
authors.initials: A
authors.lastname: Claude
authors.forename: J
authors.initials: J
authors.lastname: Gonzalez
authors.forename: R
authors.initials: R
authors.lastname: Avendaño-Herrera
authors.forename: J G
authors.initials: JG
authors.lastname: Carcamo
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001431:Bacteriological Techniques; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D060226:Head Kidney / Q000166:cytology; D053522:Piscirickettsia / Q000502:physiology*; D012474:Salmon; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D003545:Cysteine