Rhodobacter species are promising beneficial microbes that can improve growth performance, immunity and antioxidant capability in aquatic crustaceans. Yet the safety of Rhodobacter azotoformans for potential application in Chinese mitten crab Eriocheir sinensis is still unclear. In the present study, R. azotoformans SY5, a potential probiotic additive that can significantly improve the growth performance, immunity, antioxidant capability, and disease resistance in E. sinensis, was evaluated for safety through whole genome sequencing, antibiotic resistance, toxic metabolites, virulence, and crab tolerance assays. The results indicated that R. azotoformans SY5 only harboured the acyl carrier protein-encoding gene (acpP) that was universal in probiotic bacteria with the function of bacterial fatty acid biosynthesis, exhibited high susceptibility to aminoglycosides, penicillins, polymyxins, polyphosphates, quinolones, and tetracyclines antimicrobials, and possessed inability to produce hemolysin, hydrogen sulphide, nitrite, ammonia, and phenylpyruvate. In addition, R. azotoformans SY5 showed no pathogenicity for E. sinensis with the seven-day acute intraperitoneal LD₅₀ value of above 6.0 × 10⁹ cfu/ml and 30-day chronic oral LD₅₀ of above 6.0 × 10⁹ cfu/g diet. To our knowledge, this is the first report on the safety of R. azotoformans for potential application in Chinese mitten crabs.

This study investigated the impacts of white spot syndrome virus (WSSV) on the gut microbiota of Penaeus monodon through a comparative microbiota analysis of infected and healthy shrimp using 16S rDNA sequencing. The WSSV-infected shrimp exhibited characteristic white spots, reduced feeding activity, and behavioral lethargy preceding 100% mortality. The comparative microbiota analysis revealed a significantly diminished α-diversity in the infected specimens, marked by phylum-level dominance shifts from Proteobacteria (72.68%) and Firmicutes (11.27%) in the controls to Cyanobacteria (75.51%) and Proteobacteria (15.63%) in the WSSV-infected shrimp. An LEfSe analysis identified Arthrospira_PCC-7345 and Halochromatium as significantly enriched taxa during infection, contrasting with depleted populations of Ruegeria, Marivita, Bacillus, and seven other genera. The distinct dysbiosis pattern characterized by the pathogen-favored taxa proliferation and commensal species suppression demonstrates WSSV-associated microbiota restructuring, potentially contributing to disease progression in farmed P. monodon. These findings establish intestinal microbial biomarkers for early WSSV detection in aquaculture systems.