Phenotype of Aeromonas salmonicida sp. salmonicida cyclic adenosine 3′,5′-monophosphate receptor protein (Crp) mutants and its virulence in rainbow trout (Oncorhynchus mykiss)

Abstract

Precise deletion of genes related to virulence can be used as a strategy to produce attenuated bacterial vaccines. Here, we study the deletion of the cyclic-3,5-adenosine monophosphate (cAMP) receptor protein (Crp) in Aeromonas salmonicida, the aetiologic agent of furunculosis in marine and freshwater fish. The Crp protein is a conserved global regulator, controlling physiology processes, like sugar utilization. Deletion of the crp gene has been utilized in live attenuated vaccines for mammals, birds and warm water fish. Here, we characterized the crp gene and reported the effect of a crp deletion in A.salmonicida virulent and non-virulent isolates. We found that A.salmonicida crp was not able to utilize maltose and other sugars, and its generation time was similar to the wild type. A.salmonicida crp showed a higher ability of cell invasion compared to the wild type. Fish challenges showed that A.salmonicida crp is similar to 6 times attenuated in Oncorhynchus mykiss and conferred protective immunity against the intraperitoneal challenge with A.salmonicida wild type. We concluded that deletion of A.salmonicida crp influences sugar utilization, cell invasion and virulence. Deletion of crp in A.salmonicida could be considered as part of an effective strategy to develop immersion live attenuated vaccines against furunculosis.

Precise deletion of genes related to virulence can be used as a strategy to produce attenuated bacterial vaccines. Here, we study the deletion of the cyclic-3,5-adenosine monophosphate (cAMP) receptor protein (Crp) in Aeromonas salmonicida, the aetiologic agent of furunculosis in marine and freshwater fish. The Crp protein is a conserved global regulator, controlling physiology processes, like sugar utilization. Deletion of the crp gene has been utilized in live attenuated vaccines for mammals, birds and warm water fish. Here, we characterized the crp gene and reported the effect of a crp deletion in A.salmonicida virulent and non-virulent isolates. We found that A.salmonicida crp was not able to utilize maltose and other sugars, and its generation time was similar to the wild type. A.salmonicida crp showed a higher ability of cell invasion compared to the wild type. Fish challenges showed that A.salmonicida crp is similar to 6 times attenuated in Oncorhynchus mykiss and conferred protective immunity against the intraperitoneal challenge with A.salmonicida wild type. We concluded that deletion of A.salmonicida crp influences sugar utilization, cell invasion and virulence. Deletion of crp in A.salmonicida could be considered as part of an effective strategy to develop immersion live attenuated vaccines against furunculosis.