Caracterización biológica y molecular de aislados de la bacteria Piscirickettsia salmonis
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Fecha
2003
Profesor/a Guía
Facultad/escuela
Idioma
es
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Editor
Universidad Andrés Bello
Nombre de Curso
Licencia CC
Licencia CC
Resumen
Piscirickettsia salmonis es un microorganismo intracelular obligado,
responsable de las mayores pérdidas atribuidas a enfermedades bacterianas en la
salmonicultura chilena. Esta enfermedad ha presentado variaciones a través del
tiempo y es posible que presente una historia evolutiva resultante en una diversidad
filogenética, que aunque es pequeña provee una clara evidencia de diferencia entre los
aislados.
En esta tesis se caracterizaron biológica y molecularmente aislados de
P. salmonis, obtenidos desde distintos centros de cultivo de nuestro país, con el fin de
evaluar la variabilidad entre ellos.
Desde el punto de vista biológico, se observaron diferencias en las curvas de
crecimiento y en el título de los aislados. Además, se observaron diferencias en la
sensibilidad frente a los distintos antimicrobianos utilizados.
Mediante la técnica de PCR se obtuvo el producto esperado para P. salmonis en
todos los aislados. Adicionalmente, a través del análisis del ITS, a diferencia del resto
de los aislados y la cepa control, sólo el aislado PS3 presentó un patrón de migración
corto, el que se ha asociado a una menor virulencia. Además, tras utilizar las
endonucleasas de restricción, Eco RI y Pst I, PS3 al no ser digerido con Eco RI y
generar segmentos de 1050 pb y 550 pb con Pst I, fue el único aislado distinguible.
Principalmente los resultados del análisis molecular, sugieren a PS3 como una
nueva cepa de la bacteria Piscirickettsia salmonis, el que en estudios posteriores debería
ser secuenciado posibilitando el desarrollo de partidores específicos para ser utilizados
en un ensayo de PCR.
Piscirickettsia salmonis 1s the obligated intacellular microorganism, responsible of the highest losses ascribed to bacteria! diseases in the chilean fish farming industry. This disease has shown variations through the time and it is possible that presents an evolutive history resulting in phylogenetic diversity, although is modest, provides a clear evidence of differences between the isolates. In this thesis, P. salmonis isolates obtained from differents fish farming in our country, were characterized biological and molecularly in order to evaluate the variability among themselves. Through the ITS analysis, PS3 was the unique isolate that showed a short electrophoretic migration pattem which has been associated to a smaller virulence, supported to the fact that resulted sensitive to all antibiotic concentrations tested. Furthermore, after using the restriction endonucleases Eco RI and Pst I, PS3 was the only isolate distinguishable not being digested with Eco RI and producing bands of 1050 pb and 550 pb with Pst I. Mainly, the results of the molecular analysis, suggest PS3 as a new strain of P. salmonis, which in further researches should be sequenced making possible the development of specific primers to be used in a nested PCR.
Piscirickettsia salmonis 1s the obligated intacellular microorganism, responsible of the highest losses ascribed to bacteria! diseases in the chilean fish farming industry. This disease has shown variations through the time and it is possible that presents an evolutive history resulting in phylogenetic diversity, although is modest, provides a clear evidence of differences between the isolates. In this thesis, P. salmonis isolates obtained from differents fish farming in our country, were characterized biological and molecularly in order to evaluate the variability among themselves. Through the ITS analysis, PS3 was the unique isolate that showed a short electrophoretic migration pattem which has been associated to a smaller virulence, supported to the fact that resulted sensitive to all antibiotic concentrations tested. Furthermore, after using the restriction endonucleases Eco RI and Pst I, PS3 was the only isolate distinguishable not being digested with Eco RI and producing bands of 1050 pb and 550 pb with Pst I. Mainly, the results of the molecular analysis, suggest PS3 as a new strain of P. salmonis, which in further researches should be sequenced making possible the development of specific primers to be used in a nested PCR.
Notas
Tesis (Ingeniero en Acuicultura)
Palabras clave
Salmonicultura, Piscirickettsia salmonis, Enfermedades