Effect of Bacillus amyloliquefaciens and Bacillus subtilis on fermentation, dynamics of bacterial community and their functional shifts of whole-plant corn silage

Background:Bacillus amyloliquefaciens(BA)and Bacillus subtilis(BS)are usually used as feed supplements directly or bacterial inoculants in biological feeds for animals.However,few research have reported the effects of BA and BS on fermentation characteristics and bacterial community successions of whole-plant corn silage during ensiling.If the BA and BS inoculants have positive effects on silages,then they could not only improve fermentation characteristics,but also deliver BA or BS viable cells to ruminants,which would play its probiotic effect.Therefore,the objectives of this study were to investigate the effects of BA and BS on the fermentation,chemical characteristics,bacterial community and their metabolic pathway of whole-plant corn silage.Results:Freshly chopped whole-plant corn was inoculated without or with BA and BS,respectively,and ensiled for1,3,7,14 and 60 d.Results showed that BA and BS inoculations increased lactic acid concentrations of whole-plant corn silages compared with control,and BA inoculation decreased acetic acid concentrations,whereas BS inoculation decreased fiber contents and increased crude protein(CP)content.Higher water-soluble carbohydrate contents and lower starch contents were observed in BA-and BS-inoculated silages compared with that in control.The decreased CP content and increased non-protein nitrogen content were observed in BA-inoculated silage,which was consistent with the higher amino acid metabolism abundances observed in BA-inoculated silage.In addition,it was noteworthy that BA and BS inoculations increased the metabolism of cofactors and vitamins,and decreased the relative abundances of drug resistance:antimicrobial pathways.We also found that the bacterial metabolism pathways were clearly separated into three clusters based on the ensiling times of whole-plant corn silage in the present study.There were no significant differences in bacterial community compositions among the three groups during ensiling.However,BA and BS inoculations decreased the relative abundances of undesirable bacteria such as Acetobacter and Acinetobacter.Conclusion:Our findings suggested that the BS strain was more suitable as silage inoculants than the BA strain in whole-plant corn silage in this study.

Small extracellular vesicles with nanomorphology memory promote osteogenesis

Nanotopographical cues endow biomaterials the ability to guide cell adhesion, proliferation, and differentiation. Cellular mechanical memory can maintain the cell status by retaining cellular information obtained from past mechanical microenvironments. Here, we propose a new concept “morphology memory of small extracellular vesicles (sEV)” for bone regeneration. We performed nanotopography on titanium plates through alkali and heat (Ti8) treatment to promote human mesenchymal stem cell (hMSC) differentiation. Next, we extracted the sEVs from the hMSC, which were cultured on the nanotopographical Ti plates for 21 days (Ti8-21-sEV). We demonstrated that Ti8-21-sEV had superior pro-osteogenesis ability in vitro and in vivo. RNA sequencing further confirmed that Ti8-21-sEV promote bone regeneration through osteogenic-related pathways, including the PI3K-AKT signaling pathway, MAPK signaling pathway, focal adhesion, and extracellular matrix-receptor interaction. Finally, we decorated the Ti8-21-sEV on a 3D printed porous polyetheretherketone scaffold. The femoral condyle defect model of rabbits was used to demonstrate that Ti8-21-sEV had the best bone ingrowth. In summary, our study demonstrated that the Ti8-21-sEV have memory function by copying the pro-osteogenesis information from the nanotopography. We expect that our study will encourage the discovery of other sEV with morphology memory for tissue regeneration.