Abstract:
This study investigated the pathogenic bacteria of
Clarias fuscus with fulminant death in Nanning City, Pubei County and Yulin City of Guangxi, and analysed the effect of the six virulence genes on its pathogenicity. Pathogen was isolated from the heart, liver and some other parts of the diseased fish by using conventional method, then an artificial infection test was measured on the pathogenicity of the isolated strains. Identification of the isolates was confirmed by API 20NE analysis and 16S rRNA gene sequencing, including a PCR assay of detecting six kinds of virulence genes of the isolates. Results showed that five pathogenic strains were isolated from the diseased fish, including three strains of
Aeromonas hydrophila and two strains of
Aeromonas sobria. The three strains of
Aeromonas hydrophila shared the highest similarity (99.8%) with
Aeromonas hydrophila standard strain ATCC 7966 (CP000462). Meanwhile the two strains of
Aeromonas sobria shared the highest similarity (99.9%) with
Aeromonas sobria standard strain NO.106 (AB472903.1). Among the five strains, detection rates of six virulence genes were as follows. Hundred percent of the strains carried gene
Act and
Aer. Eighty percent carried gene
ahal, hly and
Alt. Only twenty percent of the strains carried gene
ahp. The five
Aeromonas strains contained three virulence genotypes in all, including the main genotype
Act+ahal+hly+Alt+ahp-Aer+ of three strains (accounting for 60% of all the strains), as well as the genotype
Act+ahal+hly+Alt+ahp+Aer+ and
Act+ahal-hly-Alt-ahp-Aer+ of one strain (each accounting for 20% of all the strains). Strain which carried all the six detected virulence genes had the highest pathogenicity, whereas the one carrying only
Act and
Aer had the minimal pathogenicity. Gene
ahp played an important role in pathogenicity of strains, and the pathogenicity of pathogenic bacteria resulted from the synergistic reaction of multiple virulence genes.