Isolation and characterization of Streptococcus agalactiae inducing mass mortalities in cultured Nile tilapia (Oreochromis niloticus) with trials for disease control using zinc oxide nanoparticles and ethanolic leaf extracts of some medicinal plants.

Abstract

Background: Streptococcus agalactiae (Group B streptococcus, GBS) induces a serious infection that can harm not only aquatic life but also humans and other animals. In a fish farm in southern Egypt, Nile tilapia (Oreochromis niloticus) has developed an epidemic with clinical symptoms resembling piscine streptococcosis.

Results: Initial microscopic inspection of the affected fish brain and kidney indicated the presence of Gram-positive cocci. S. agalactiae was effectively isolated and identified using nucleotide homology of the 16S rRNA and species-specific PCR. The partial 16S rRNA sequence was deposited in the GenBank database at the NCBI and given the accession number MW599202. Genotyping using RAPD analysis indicated that the isolates in the present study belonged to the same genotypes and had the same origin. The challenge test, via immersion (9.2 × 10⁷, 9.2 × 10⁶, and 9.2 × 10⁵ CFU/ml for 1 h) or intraperitoneal injection (4.6 × 10⁷, 4.6 × 10⁶, and 4.6 × 10⁵ CFU/fish), elicited clinical symptoms resembling those of naturally infected fish with a mortality rate as high as 80%. The ability to create a biofilm as one of the pathogen virulence factors was verified. Zinc oxide nanoparticles and the ethanolic leaf extracts of nine medicinal plants demonstrated considerable antibacterial activities against the tested S. agalactiae strain with low minimum bactericidal concentrations (MBC) and minimum inhibitory concentrations (MIC). The ethanolic leaf extracts from Lantana camara and Aberia caffra showed potent antibacterial activity with MBC values of 0.24 and 0.485 mg/ml, and MIC values of 0.12 & 0.24 mg/ml, respectively.

Conclusion: This study isolated S. agalactiae from O. niloticus mortalities in a fish farm in Assiut, Egypt. The pathogen persists in fish environments and can escape through biofilm formation, suggesting it cannot be easily eliminated. However, promising findings were obtained with in vitro control employing zinc oxide nanoparticles and medicinal plant extracts. Nevertheless further in vivo research is needed.