异常毕赤酵母和纤维素酶对青贮甜高粱产甲烷潜力的影响

Effects of Pichia anomala and cellulase on methane production potential of sweet sorghum silage

  • 摘要: 本文探究了单独或联合使用异常毕赤酵母(Pichia anomala)和纤维素酶对甜高粱青贮质量和厌氧消化产甲烷潜力的调控效果, 利用高通量测序解析了厌氧消化过程中微生物菌群的多样性, 结合经济性能分析筛选廉价高效的青贮预处理添加剂。结果表明, 甜高粱青贮过程中, 单独或联合使用2种添加剂的青贮品质均为优良, 其中异常毕赤酵母和纤维素酶联合处理组(PC组)的综合评价值为0.66, 单独添加异常毕赤酵母(Pa组)的评价值次之为0.63; 两种添加剂均能有效保存青贮甜高粱的可溶性碳水化合物、粗蛋白、纤维素和半纤维素等能量组分, 降低酸性洗涤木质素、中性洗涤纤维和酸性洗涤纤维等木质纤维组分, 增加乳酸和乙酸含量, 强化青贮发酵进程。尤其, PC组的可溶性碳水化合物保存效果最佳, 木质素去除率高达62.55%; Pa组中蛋白质保存完好, 乳酸和乙酸含量最高, 分别为50.01 g·kg−1(DM)和18.35 g·kg−1(DM)。青贮预处理能明显提升甜高粱的厌氧发酵产甲烷效能, 与新鲜原料组相比, PC组累计产甲烷量提高30.13%, 为457.70 mL(CH4)·g−1(VS), 消化迟滞期缩短62.96%, 生物降解指数最高, 达72.39%; Pa组的预处理效果次之, 最大产甲烷速率提升33.57%, 消化迟滞期缩短33.33%, 且消化系统都相对稳定。厌氧消化系统中属水平优势细菌为发酵单胞菌(Fermentimonas)和梭状芽孢杆菌(Clostridium_sensu_stricto_1), 与pH呈负相关, 与化学需氧量(COD)和总挥发性脂肪酸(TVFA)呈正相关。优势古菌属为甲烷八叠球菌(Methanosarcina)和甲烷短杆菌(Methanobrevibacter), 其中甲烷八叠球菌与COD和TVFA呈负相关, 与pH呈正相关; 甲烷短杆菌与COD和TVFA呈正相关, 与pH呈负相关关系。综合青贮质量、产甲烷性能和经济性分析, 推荐单独使用异常毕赤酵母作为甜高粱青贮预处理的生物强化剂。

     

    Abstract: The effects of Pichia anomala, cellulase, and a combination of both on the regulation of ensiling quality and methanogenic potential during anaerobic digestion of sweet sorghum were investigated in this study. Moreover, microbial community diversity during anaerobic digestion was analyzed using high-throughput DNA sequencing, and economic performance was evaluated to screen for inexpensive and highly efficient additives. The results revealed that the two additives improved the ensiling fermentation quality of sweet sorghum to different extents. The highest comprehensive assessment value was for the silages treated with the combination of P. anomala and cellulase (PC, 0.66), followed by 0.63 in silages treated with P. anomala alone (Pa). PC was effective in preserving energy components such as water-soluble carbohydrates, crude protein, cellulose, and hemicellulose in sweet sorghum silage. The addition of the two could also reduce lignocellulosic components, such as acid detergent lignin, neutral detergent fiber, and acid detergent fiber and subsequently increase the content of lactic and acetic acid and enhance ensiling fermentation. In particular, there were more residual water-soluble carbohydrates and the highest lignin removal (62.55%) after PC treatment, and the well-preserved protein and the highest lactic and acetic acid content, 50.01 g·kg1 and 18.35 g·kg−1 (based on dry matter, DM), were determined in Pa silages. Ensiling pretreatment markedly improved the methanogenic potential of sweet sorghum. In particular, for PC, the maximum cumulative methane production was 457.70 mL(CH4)·g1 (based on volatile solids, VS), which was increased by 30.13% compared to raw sweet sorghum, the maximum biodegradability index was 72.39%, and the lag phase was decreased by 62.96% compared to raw sweet sorghum (CK). In comparison with CK, the maximum methane production rate in Pa increased by 33.57%, and the lag phase decreased by 33.33%. The species richness of bacteria and archaea increased after sweet sorghum was treated with additives. Simultaneously, the use of different silage additives can affect the variation in bacterial community diversity with fermentation time but no such effect was observed in archaea. At the genus level, the dominant bacteria in the anaerobic digestion effluent were Fermentimonas and Clostridium_ sensu_stricto_1, which negatively correlated with pH and positively correlated with chemical oxygen demand (COD) and total volatile fatty acid (TVFA). The dominant archaea were Methanosarcina and Methanobrevibacter, where Methanosarcina was negatively correlated with COD and TVFA and positively correlated with pH. Methanobrevibacter was positively correlated with COD and TVFA concentrations and negatively correlated with pH. After the combined analysis of ensiling quality, methanogenic performance, and economy, it is recommended to use Pa as a biological additive for ensiling pretreatment in practical production.

     

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