Comparison of the effects of rumen-protected and unprotected L-leucine on fermentation parameters, bacterial composition, and amino acids metabolism in in vitro rumen batch cultures

This study was conducted to compare the effects of rumen-protected (RP-Leu) and unprotected L-leucine (RU-Leu) on the fermentation parameters, bacterial composition, and amino acid metabolism in vitro rumen batch incubation. The 5.00 g RP-Leu or RU-Leu products were incubated in situ in the rumen of four beef cattle (Bos taurus) and removed after 0, 2, 4, 6, 12, 16, and 24 h to determine the rumen protection rate. In in vitro incubation, both RP-Leu and RU-Leu were supplemented 1.5 mmol/bottle (L-leucine HCl), and incubated after 0, 2, 4, 6, 8, 12, and 16 h to measure gas production (GP), nutrient degradability, fermentation parameters, bacterial composition, and amino acids metabolism. Results from both in vitro and in situ experiments confirmed that the rumen protection rate was greater (p < 0.01) in RP-Leu than in RU-Leu, whereas the latter was slow (p < 0.05) degraded within incubation 8 h. Free leucine from RP-Leu and RU-Leu reached a peak at incubation 6 h (p < 0.01). RU-Leu supplementation increased (p < 0.05) gas production, microbial crude protein, branched-chain AAs, propionate and branched-chain VFAs concentrations, and Shannon and Sobs index in comparison to the control and RP-Leu supplementation. RU-Leu and RP-Leu supplementation decreased (p < 0.05) the relative abundance of Bacteroidota, which Firmicutes increased (p < 0.05). Correlation analysis indicated that there are 5 bacteria at the genus level that may be positively correlated with MCP and propionate (p < 0.05). Based on the result, we found that RP-Leu was more stable than RU-Leu in rumen fluid, but RU-Leu also does not exhibit rapid degradation by ruminal microbes for a short time. The RU-Leu was more beneficial in terms of regulating rumen fermentation pattern, microbial crude protein synthesis, and branched-chain VFAs production than RP-Leu in vitro rumen conditions

Remediation Performance and Mechanism of Heavy Metals by a Bottom Up Activation and Extraction System Using Multiple Biochemical Materials

Soil contamination with heavy metals has caused serious environmental problems and increased the risks to humans and biota. Herein, we developed an effective bottom up metals removal system based on the synergy between the activation of immobilization metal-resistant bacteria and the extraction of bioaccumulator material (<i>Stropharia rugosoannulata</i>). In this system, the advantages of biochar produced at 400 °C and sodium alginate were integrated to immobilize bacteria. Optimized by response surface methodology, the biochar and bacterial suspension were mixed at a ratio of 1:20 (w:v) for 12 h when 2.5% sodium alginate was added to the mixture. Results demonstrated that the system significantly increased the proportion of acid soluble Cd and Cu and improved the soil microecology (microbial counts, soil respiration, and enzyme activities). The maximum extractions of Cd and Cu were 8.79 and 77.92 mg kg^–1, respectively. Moreover, details of the possible mechanistic insight into the metal removal are discussed, which indicate positive correlation with the acetic acid extractable metals and soil microecology. Meanwhile, the “dilution effect” in <i>S. rugosoannulata</i> probably plays an important role in the metal removal process. Furthermore, the metal-resistant bacteria in this system were successfully colonized, and the soil bacteria community were evaluated to understand the microbial diversity in metal-contaminated soil after remediation