Comparative in silico and in vivo study of the antioxidant activity of lactoferrin, Geobacillus stearothermophilus, and Lactobacillus delbrueckii subsp. lactis against Rotavirus infection in male mice

Abstract Rotavirus is a major cause of pediatric gastroenteritis, for which effective treatments are limited. This study investigates the antioxidant and antiviral potential of lactoferrin, Geobacillus stearothermophilus, and Lactobacillus delbrueckii subsp. lactis against Rotavirus infection. In this study, Geobacillus stearothermophilus and Lactobacillus delbrueckii subsp. lactis were isolated from Hammam Pharon soil and milk cheese, respectively, and identified using molecular techniques with accession numbers PP758390 and PP758383. The antioxidant effect against DPPH showed that lactoferrin exhibited the strongest scavenging ability, followed by Geobacillus stearothermophilus and Lactobacillus delbrueckii subsp. lactis. In vivo experiments involved administering lactoferrin, Geobacillus stearothermophilus, and Lactobacillus delbrueckii subsp. lactis in the drinking water of young mice for three days, followed by Rotavirus infection on the fourth day and sacrifice on the fifth day. The results demonstrated that lactoferrin significantly reduced the pathogenic effects of Rotavirus, as indicated by the normalization of inflammatory cytokines (TNF-α and IL-6) in the serum (p ≤ 0.001). Histological examination of small intestinal sections from Rotavirus-infected mice revealed extensive destruction of villus structures, while mice treated with lactoferrin showed no pathological changes compared to the control group. Geobacillus stearothermophilus-treated mice exhibited less pathological alteration and Lactobacillus delbrueckii subsp. lactis-treated mice showed mild pathological changes. Additionally, molecular docking studies indicated that bacteriocin (a bacterial protein) exhibited the highest binding affinity for the Rotavirus outer membrane protein (VP6) at -261.92 kcal/mol, outperforming lactoferrin (-229.32 kcal/mol). Additionally, bacteriocin's active compounds, turimicin (-7.9 kcal/mol) and lactin (-6.5 kcal/mol), also showed strong binding to VP6, suggesting their potential as therapeutic agents against Rotavirus. In conclusion, this study highlights the significant antiviral potential of lactoferrin against Rotavirus, demonstrating its ability to mitigate pathological changes and normalize inflammatory responses in infected mice. The findings also suggest that bacteriocins, particularly those with high binding affinities to Rotavirus proteins, could serve as promising candidates for therapeutic interventions against Rotavirus infections.