Insight of microbial interactions in silage inoculation for amending fermentation quality and antioxidant capacity based on combination of modern multi-sensor technology, molecular microbiological methods, and optimal selection modelling

青贮饲料是世界范围内农场反刍动物最重要的饲料和沼气（即生物能源生产）的基质。在过去的三十年里，各种生物/化学添加剂被应用于改善青贮饲料的发酵品质和抗氧化能力。为了更深入地了解生物青贮添加剂中微生物的作用，了解青贮接种过程中微生物的相互作用是至关重要的，重点是优化青贮和乳酸菌（LAB）的优化选择。该方案提出了一个跨学科的基础研究，结合多传感器融合测量，分子微生物学分析，最优控制理论下的生化LAB发酵和普通青贮作物LAB的最佳光照。波恩大学的两个研究小组将合作进行这项研究：一个由W。Büscher负责使用多传感器技术原位筛选微生物活性，其中一个由A. Lipski负责分子微生物学解释。本研究的亮点可以概括为以下几个方面：●两个团队都对LAB菌株的最佳预选和LAB发酵的最佳化进行了初步研究（新的实验方法和首次报道的实验数据）。●一系列先进的多传感器仪器，可在厌氧、准好氧和好氧实验条件下，对青贮饲料中的CO2、乙醇、O2、pH和温度进行原位测量，确保在好氧实验中精确跟踪/筛选乳酸菌厌氧发酵过程中的微生物活动/代谢和青贮饲料中乳酸的好氧分解过程。 ●与Bolza方程（应用数学中最优控制理论的基本方程）的广义问题相关的数学建模，并且进行所提议的实验将导致最优的日照和LAB菌株的选择。●对来自青贮饲料的分离物进行共培养分析，以检测青贮期间菌株之间的互利相互作用。将分析单一菌株和共培养物的生长速率和产酸量。为了表征相互作用，将分析群体感应自诱导物的合成和作用以及共培养物的转录组分析，以检测调节相互作用。通过所开发的多传感器微型生物反应器系统，将对共培养物的互惠活动进行量化和详细分析。

Silage is worldwide the most important feedstuff for farm ruminants and substrate for biogas i.e. bioenergy production. Various biological/chemical additives have been applied for both amending fermentation quality and antioxidant capacity of silage over the last three decades. For deeper understanding of microbial roles of biological silage additives, it is critical to get insight of microbial interaction in silage inoculation, focused on optimal ensiling and optimal selection of lactic acid bacteria (LAB). This proposal presents an interdisciplinary fundamental study, combined with multi-sensor-fusion measurement, molecular microbiological analyses, biochemical LAB fermentation under optimal control theory and optimal insolation of LAB from common silage crops. Two research teams of Bonn University will cooperate together for the proposed study: one leaded by Prof. Dr. W. Büscher responsible for in situ screening of microbial activities by using multi-sensor technology and one leaded by Prof. Dr. A. Lipski responsible for molecular microbiological interpretation. The highlights of this study can be characterized as follows: ● The sounded works of preliminary study (novel experimental methods and first reported experimental data) about the optimal preselection of the strains of LAB and optimal LAB fermentation are presented from both teams. ●A series of advanced multi-sensor instruments for in situ measurement of CO2, Ethanol, O2, pH and temperature from or within silage, under anaerobic, semi-aerobic and aerobic experimental conditions, this ensures to precisely track/screen microbial activities/ metabolism during the process of LAB anaerobic fermentation and the aerobic decomposition of lactic acid in silage during aerobic experiment. ● Mathematical modelling associated with the generalized problem of Bolza equation (a basic equation of optimal control theory in applied mathematics), and to conduct the proposed experiment will result in optimal insolation and selection of LAB’s strains. ● The analyses of co-cultures of isolates from silage to detect mutualistic interactions between strains during ensiling biochemical activities. Growth rates and acid production will be analyzed for single strains and co-cultures. For characterizing the interactions the synthesis and effects of quorum sensing autoinducers will be analyzed as well as transcriptome analyses of co-cultures to detect regulatory interactions. The mutualistic activities of co-cultures will be quantified and analyzed in detail by means of the developed multi-sensor mini bioreactor system.