School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, China.
State Key Laboratory of Microbial Metabolism, School of Life Science & Biotechnology, Shanghai Jiao Tong Universitygrid.16821.3c, Shanghai, China.
Microbiol Spectr. 2022 Apr 27;10(2):e0067222. doi: 10.1128/spectrum.00672-22. Epub 2022 Mar 21.
Probiotics have been widely used to prevent primary Clostridioides difficile infection (pCDI); however, there are fewer studies on their therapeutic aspects for pCDI. In this study, high doses of Bifidobacterium breve YH68 were used alone or in combination with vancomycin (VAN) and metronidazole (MTR) to treat pCDI mice. Mouse feces were collected from preinfection, postinfection, and posttreatment stages. Subsequently, the C. difficile number and toxin level in feces were detected by plate count method and C. difficile toxin enzyme-linked immunosorbent assay (ELISA). Simultaneously, 16S rRNA amplicon sequencing and untargeted metabolomics were employed to explore the changing patterns and characteristic markers of fecal microbiota and metabolome. The results indicated that high doses of YH68 used alone or in combination with VAN and MTR were more effective than the combination of VAN and MTR for pCDI mice and improved their final survival rate. This probiotic strain and its combination with antibiotics reduced C. difficile numbers and toxin levels in the feces, downregulated proinflammatory cytokine levels in colon tissue, and alleviated cecum tissue hyperplasia. Meanwhile, the level of fecal microbiota diversity increased significantly in pCDI mice after treatment, with an increase in the relative abundance of , , , unidentified, and , and this process was accompanied by elevated levels of secondary bile acid, butyric acid, and gentamicin C1a and reduced levels of primary bile acid and indoles. Most notably, the combination of YH68 with VAN and MTR diminished the damaging effect of antibiotic treatment alone on the microbiota. Our findings suggested that high doses of YH68 used in combination with VAN and MTR have a better therapeutic effect on pCDI mice than the combination of VAN and MTR alone. Many studies have focused on the preventive effects of probiotics against pCDI, but few studies have investigated in depth the therapeutic effects of probiotics, especially at the postinfection stage. We demonstrated that high doses of Bifidobacterium breve YH68 used alone or in combination with vancomycin (VAN) and metronidazole (MTR) exerted outstanding efficacy in the treatment of pCDI mice. This probiotic-antibiotic combination regimen has the potential to be a new option for the clinical treatment of pCDI.
益生菌被广泛用于预防原发性艰难梭菌感染（pCDI）；然而，关于其治疗 pCDI 的研究较少。在这项研究中，单独使用或联合万古霉素（VAN）和甲硝唑（MTR）使用高剂量短双歧杆菌 YH68 治疗 pCDI 小鼠。从小鼠感染前、感染后和治疗后阶段收集粪便。随后，采用平板计数法和艰难梭菌毒素酶联免疫吸附试验（ELISA）检测粪便中艰难梭菌数量和毒素水平。同时，采用 16S rRNA 扩增子测序和非靶向代谢组学方法探索粪便微生物群和代谢组的变化模式和特征标志物。结果表明，与 VAN 和 MTR 联合使用相比，YH68 单独或联合使用对 pCDI 小鼠更有效，提高了其最终存活率。该益生菌株及其与抗生素的联合使用降低了粪便中的艰难梭菌数量和毒素水平，下调了结肠组织中的促炎细胞因子水平，并缓解了盲肠组织增生。同时，pCDI 小鼠治疗后粪便微生物群多样性水平显著升高，相对丰度增加， ， ，未鉴定，和 ，这一过程伴随着次级胆汁酸、丁酸和庆大霉素 C1a 水平的升高，以及初级胆汁酸和吲哚水平的降低。值得注意的是，YH68 与 VAN 和 MTR 的联合使用减轻了抗生素单独治疗对微生物群的破坏性影响。我们的研究结果表明，与 VAN 和 MTR 联合使用高剂量 YH68 对 pCDI 小鼠的治疗效果优于 VAN 和 MTR 联合使用。许多研究集中在益生菌对 pCDI 的预防作用上，但很少有研究深入探讨益生菌的治疗作用，尤其是在感染后阶段。我们证明，单独使用或联合万古霉素（VAN）和甲硝唑（MTR）使用高剂量短双歧杆菌 YH68 对 pCDI 小鼠具有显著疗效。这种益生菌-抗生素联合方案有可能成为治疗 pCDI 的新选择。
