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

A. J. Yañez, K. Valenzuela, H. Silva, J. Retamales, A. Romero, R. Enriquez, J. Figueroa, A. Claude, J. Gonzalez, R. Avendaño-Herrera, J. G. Carcamo

Resultado de la investigación: Article

43 Citas (Scopus)

Resumen

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 cellculture 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 AUSTRALSRS 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.

Idioma originalEnglish
Páginas (desde-hasta)197-205
Número de páginas9
PublicaciónDiseases of Aquatic Organisms
Volumen97
N.º3
DOI
EstadoPublished - 24 ene 2012

Huella dactilar

Piscirickettsia salmonis
salmonid
fish culture
pathogen
culture media
pathogens
fish
apoptosis
septicemia
vaccine
virulence
absorbance
purification
incubation
membrane
DNA
kinetics
liquid
protein
cytopathogenicity

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Aquatic Science

Citar esto

Yañez, A. J., Valenzuela, K., Silva, H., Retamales, J., Romero, A., Enriquez, R., ... Carcamo, J. G. (2012). Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis. Diseases of Aquatic Organisms, 97(3), 197-205. https://doi.org/10.3354/dao02403
Yañez, A. J. ; Valenzuela, K. ; Silva, H. ; Retamales, J. ; Romero, A. ; Enriquez, R. ; Figueroa, J. ; Claude, A. ; Gonzalez, J. ; Avendaño-Herrera, R. ; Carcamo, J. G. / Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis. En: Diseases of Aquatic Organisms. 2012 ; Vol. 97, N.º 3. pp. 197-205.
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Yañez, AJ, Valenzuela, K, Silva, H, Retamales, J, Romero, A, Enriquez, R, Figueroa, J, Claude, A, Gonzalez, J, Avendaño-Herrera, R & Carcamo, JG 2012, 'Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis', Diseases of Aquatic Organisms, vol. 97, n.º 3, pp. 197-205. https://doi.org/10.3354/dao02403

Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis. / Yañez, A. J.; Valenzuela, K.; Silva, H.; Retamales, J.; Romero, A.; Enriquez, R.; Figueroa, J.; Claude, A.; Gonzalez, J.; Avendaño-Herrera, R.; Carcamo, J. G.

En: Diseases of Aquatic Organisms, Vol. 97, N.º 3, 24.01.2012, p. 197-205.

Resultado de la investigación: Article

TY - JOUR

T1 - Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis

AU - Yañez, A. J.

AU - Valenzuela, K.

AU - Silva, H.

AU - Retamales, J.

AU - Romero, A.

AU - Enriquez, R.

AU - Figueroa, J.

AU - Claude, A.

AU - Gonzalez, J.

AU - Avendaño-Herrera, R.

AU - Carcamo, J. G.

PY - 2012/1/24

Y1 - 2012/1/24

N2 - 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 cellculture 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 AUSTRALSRS 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.

AB - 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 cellculture 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 AUSTRALSRS 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.

KW - Broth

KW - Culture

KW - Piscirickettsia salmonis

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U2 - 10.3354/dao02403

DO - 10.3354/dao02403

M3 - Article

C2 - 22422090

AN - SCOPUS:84856188698

VL - 97

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EP - 205

JO - Diseases of Aquatic Organisms

JF - Diseases of Aquatic Organisms

SN - 0177-5103

IS - 3

ER -

Yañez AJ, Valenzuela K, Silva H, Retamales J, Romero A, Enriquez R y otros. Broth medium for the successful culture of the fish pathogen Piscirickettsia salmonis. Diseases of Aquatic Organisms. 2012 ene 24;97(3):197-205. https://doi.org/10.3354/dao02403