Microbial Identification and Population Successions in the Novel Carabao Hydrolysis Pretreatment of Agriculture Crops Lignocelluloses Intended for Cellulose Ethanol Production

P. Florendo
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Abstract

This study was conducted with the objective to elucidate physical, chemical and microbial changes in carabao rumen fluid hydrolysis of bagasse of sweet sorghum and sugarcane, corn stover and rice straw. Identification of microbial species, composition of populations and succession behavior at durations of 3, 6, and 9days of the carabao novel process were determined using API20A kit for anaerobes, spectrophotometry and compound microscopy. Efficiency of microbial conversion of carbohydrates into soluble sugars, pH changes in hydrolysates were determined at various durations. Morphology and physiochemical characterizations showed the occurrences of bacterial groups Clostridium, Bacteroidetes, Streptococcus, Actinomyces, Bifidobacterium, Lactobacillus and Staphylococcus, rumen fungi species Ruminomyces, Orpinomyces and Neocallimastix and various protozoa of the family Ophryoscolocedae and Isotrichia. Population of microbes was highest in the hydrolysate crop residues sweet sorghum, corn stover, sugarcane and rice straw with corresponding significant difference in average carbohydrates conversion efficiency of 66.49%, 52.43% 52.12%, and 39.28% respectively(p<0.05). Novel process with duration of 6 days had the highest carbohydrates conversion efficiency average of 55.02%. Hydrolysis initial average pH 6.98 had turned acidic at durations of 3 days, 6 days and 9 days while pH variations were significantly different among feedstock, lowest in sugarcane(pH 4.91) sweet sorghum(pH 5.46), corn stover (pH 5.72) and near neutral in rice straw(pH 6.56). Succession behavior of bacteria, fungi and protozoan had influenced microbial population, growth pattern and diversity of species in the novel carabao. In conclusion, the isolation, morphological and physiochemical characterizations of rumen bacteria, fungi and protozoa shows the diversity in the microbial community in the novel carabao, has provided elucidation of the novel carabao microbes efficient conversion of carbohydrates into soluble sugars. Information generated in this study will be of great help in understanding the novel carabao and the improvements when the process proceeds to upscaling as pretreatment of lignocellulosic feedstocks intended for cellulose ethanol production.
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新型卡拉宝水解预处理农作物木质纤维素用于纤维素乙醇生产的微生物鉴定和种群演替
本研究旨在阐明甜高粱甘蔗渣、甘蔗甘蔗渣、玉米秸秆和水稻秸秆水解后卡拉宝瘤胃液的物理、化学和微生物变化。采用API20A厌氧菌检测试剂盒、分光光度法和复合显微镜对新工艺3、6和9d的微生物种类鉴定、种群组成和演替行为进行了测定。测定了微生物将碳水化合物转化为可溶性糖的效率,以及不同时间下水解产物的pH变化。形态学和理化特征显示,有梭状芽孢杆菌、拟杆菌门、链球菌、放线菌门、双歧杆菌门、乳杆菌门、葡萄球菌门等菌群,瘤胃真菌类有Ruminomyces、Orpinomyces和Neocallimastix,以及各种原生动物有ophophcolocedae科和等毛菌科。水解作物秸秆中微生物数量最多的是甜高粱、玉米秸秆、甘蔗和水稻秸秆,平均碳水化合物转化效率分别为66.49%、52.43%、52.12%和39.28%,差异显著(p<0.05)。新工艺持续时间为6 d,碳水化合物转化效率最高,平均为55.02%。水解初始平均pH为6.98,在3天、6天和9天时呈酸性,不同原料的pH变化差异显著,甘蔗最低(pH 4.91),甜高粱最低(pH 5.46),玉米秸秆最低(pH 5.72),水稻秸秆接近中性(pH 6.56)。细菌、真菌和原生动物的演替行为影响了新型卡拉宝的微生物种群、生长模式和物种多样性。综上所述,瘤胃细菌、真菌和原生动物的分离、形态和理化特征显示了新型卡拉宝微生物群落的多样性,为阐明新型卡拉宝微生物将碳水化合物高效转化为可溶性糖提供了依据。本研究产生的信息将极大地帮助我们了解这种新型的carabao,以及当该工艺升级为用于纤维素乙醇生产的木质纤维素原料的预处理时的改进。
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