Isolation and characterization of infectious Vibrio parahaemolyticus, the causative agent of AHPND, from the whiteleg shrimp (Litopenaeus vannamei)

Antonio Luna González, Patricia López León, Ruth Escamilla Montes, Ma. del Carmen Flores Miranda, Jesús A. Fierro Coronado, Píndaro Álvarez Ruiz, Genaro Diarte Plata


DOI: https://doi.org/10.3856/vol44-issue3-fulltext-5

Abstract


Vibrio parahaemolyticus, the causative agent of acute hepatopancreatic necrosis disease (AHPND), was isolated from the hepatopancreas of moribund whiteleg shrimp of commercial farms (Guasave, Sinaloa, Mexico). The isolates were screened on thiosulfate citrate bile salt sucrose agar plates for the selection of green colonies and further characterized through PCR with primers AP3, 89F/R, hemolysin genes, hemolytic and enzymatic activity, hydrophobicity, autoaggregation, and biofilm formation. Immersion experimental infections were conducted to confirm the pathogenicity of selected bacterial strains. In addition, the LC50 was calculated for each isolate. All isolates (35) belonged to V. parahaemolyticus, but three isolates did not correspond to strains that cause AHPND since they were negative with primers 89F/R. All isolates were α-hemolytic and showed biofilm formation (from moderate to strong). Isolates were hydrophobic or hydrophilic and showed high autoaggregation capacity. Nine strains did not kill shrimp and eleven were pathogenic, but differences in virulence were found among them perhaps due to α-hemolysis and differences in biofilm formation and hydrophobicity. Results showed that smaller shrimp are less resistant to immersion bacterial infection. The studied virulence factors may help to understand the pathogenicity of V. parahaemolyticus.


Luna González A, López León P, Escamilla Montes R, Flores Miranda M, Fierro Coronado J, Álvarez Ruiz P, Diarte Plata G. Isolation and characterization of infectious Vibrio parahaemolyticus, the causative agent of AHPND, from the whiteleg shrimp (Litopenaeus vannamei). Lat. Am. J. Aquat. Res.. 2017;44(3). Available from: doi:10.3856/vol44-issue3-fulltext-5 [Accessed 22 Jan. 2019].
Luna González, A., López León, P., Escamilla Montes, R., Flores Miranda, M., Fierro Coronado, J., Álvarez Ruiz, P., & Diarte Plata, G. (2017). Isolation and characterization of infectious Vibrio parahaemolyticus, the causative agent of AHPND, from the whiteleg shrimp (Litopenaeus vannamei). Latin American Journal of Aquatic Research, 44(3). doi:http://dx.doi.org/10.3856/vol44-issue3-fulltext-5