Foodborne diseases have been witnessing a constant rising trend worldwide, mainly caused by pathogenic microorganisms, such as Bacillus spp., posing a direct threat to public health. The purpose of this study was to evaluate the biological risk of foodborne and probiotic Bacillus spp. in Beijing markets. A total of 55 Bacillus isolates, including 29 B. cereus, 9 B. licheniformis and 7 B. subtilis, mostly found in dairy products (32.7%), were recovered from 106 samples and identified by matrix-assisted laser desorption/ionization mass spectrometry and polymerase chain reaction methods. The susceptibility towards 16 antibiotics was determined using a broth microdilution method. Bacillus showed a high level of resistance to florfenicol (100%), lincomycin (100%), tiamulin (78.2%) and ampicillin (67.3%), while they were all susceptible or intermediate to vancomycin and rifampin. Additionally, we obtained the whole genome of 19 Bacillus strains using high-throughput sequencing, and the rates of resistance genes van, fosB, erm and tet were 57.9%, 57.9%, 21.1% and 26.3%, respectively. Moreover, 100%, 9.1%, 45.5% and 100% of these isolates carried virulence genes nhe, hbl, cytK and entFM, respectively. Lastly, 60% Bacillus strains were positive in hemolysis tests, and 3 B. licheniformis strains displayed an inhibitory activity on the growth of S. aureus ATCC 29213 using agar overlay technique. Our study outlines the characteristics of foodborne Bacillus spp. and provides information for the monitoring of food safety.

The increasing incidence of multidrug-resistant (MDR) Salmonella enterica serovar Typhimurium (S. Tm), known for causing invasive enteric infections, presents a significant public health challenge. Given the diminishing efficacy of existing antibiotics, it is imperative to explore novel alternatives for the treatment of MDR S. Tm infections. Here, we identified esculetin (EST), a natural coumarin abundant in dietary foods and herbs, as a compound exhibiting broad-spectrum antibacterial properties against a range of MDR bacteria. Our findings demonstrate that EST effectively inhibited the proliferation and expansion of MDR S. Tm in both in vitro experiments and animal models. Specifically, EST significantly downregulated the type 3 secretion system-1 (T3SS-1) virulence expression of MDR S. Tm, thereby preventing its invasion into intestinal epithelial cells. In S. Tm-infected mice, we observed cecal injury characterized by the upregulation of inflammatory cytokines, a reduction in goblet cell numbers, a decreased expression of tight junction proteins, and microbial dysbiosis. Conversely, EST treatment ameliorated these pathological changes induced by S. Tm infection and reduced oxidative stress by activating the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway, thereby improving intestinal barrier function. These results suggest that dietary coumarins or a targeted plant-based diet may offer a promising strategy to counteract MDR bacteria-induced enteric diseases.

Probiotics have been associated with clinical infections, toxicity, and antimicrobial resistance transfer, raising public concerns. Probiotic enterococci are emerging food risks as opportunistic pathogens, yet little attention has been paid to them. Herein, we collected 88 enterococcal isolates from probiotic products used for humans, companion animals, livestock, and aquaculture. Results showed that all 88 probiotic enterococcal isolates harbored diverse virulence genes, multiple antimicrobial resistance genes, and mobile genetic elements. Notably, 77 isolates were highly resistant to gentamicin. Representative enterococcal isolates exerted toxic activities in both in vitro and in vivo models. Collectively, our findings suggest that probiotic enterococci may be harmful to hosts and pose a potential threat to public health.