糖厂滤泥堆肥发酵过程中可培养芽孢杆菌种群动态变化研究
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摘要
为了揭示糖厂滤泥堆肥发酵生产生物有机肥过程中芽孢杆菌种群变化动态,本研究以糖厂滤泥堆肥发酵加工不同阶段的样品为研究对象,对各样品中的芽孢杆菌进行分离鉴定,研究糖厂滤泥堆肥发酵过程中芽孢杆菌种群数量和结构的变化动态,并分析其与本土芽孢杆菌的关系。结果表明,糖厂滤泥堆肥发酵加工过程中芽孢杆菌种类比较丰富,从糖厂滤泥堆肥发酵加工6个阶段样品中分离到64株芽孢杆菌,分属于7个属27种,其中芽孢杆菌属占绝对优势,有16种;研究发现有6种芽孢杆菌为我国新纪录种,它们是:大豆发酵芽孢杆菌(Bacillus glycinifermentans FJAT-47935、FJAT-47944、FJAT-47907)、外村尚芽孢杆菌(Bacillus hisashii FJAT-47926)、科研中心哈格瓦氏菌(Bhargavaea cecembensis FJAT-47948)、苯乙酮赖氨酸芽孢杆菌(Lysinibacillus acetophenoni FJAT-47912、FJAT-47934、FJAT-47919)、巴伐利亚鸟氨酸芽孢杆菌(Ornithinibacillus bavariensis FJAT-47915)和拾蛤鸟氨酸芽孢杆菌(Ornithinibacillus scapharcae FJAT-47951)。在糖厂滤泥不同发酵加工阶段,芽孢杆菌种类结构不同,堆肥发酵初期(1 d)以大豆发酵芽孢杆菌(Bacillus glycinifermentans FJAT-47935)为优势种,总含量为114万cfu·g-1,发酵中期(30 d)以蓼蓝大洋芽孢杆菌(Oceanobacillus polygoni FJAT-47932)为优势种,总含量为237万cfu·g-1,发酵后期(60 d)以淤泥大洋芽孢杆菌(Oceanobacillus caeni FJAT-47952)为优势种,总含量为37.6万cfu·g-1,加工成产品后以耐温芽孢杆菌(Bacillus thermotolerans FJAT-47921)为优势种,总量为43万cfu·g-1;可培养芽孢杆菌种类和数量差异显著,数量变化范围在-80%～+100%;种类的变更率在62%～84%。本土芽孢杆菌对糖厂滤泥堆肥发酵的影响较小,其发酵过程中出现的芽孢杆菌主要来源于自带种类,因此,糖厂滤泥堆肥发酵全过程不外加芽孢杆菌接种剂,自带的芽孢杆菌就能发挥作用。综上所述,糖厂滤泥堆肥发酵加工过程中不仅有种类丰富的芽孢杆菌,而且其种群数量和结构差异显著。
The aim of this study is to determine dynamic changes in Bacillus-like species population in the process of sugar-refinery filtering mud composting fermentation to produce bio-organic fertilizers. The Bacillus-like species in the samples at different composting andprocessing stages were isolated and identified to analyze the dynamic changes in their population size and structure during the process, and to reveal their relationship with the indigenous Bacillus-like species. The results indicated that the Bacillus-like species were enriched in the process of sugar-refinery filtering mud composting fermentation. A total of 64 strains, belonging to 27 species within 7 genera, were isolated from 6 samples at different composting and processing stages. Among them, the genus Bacillus, including 16 species, was the absolutely dominant population. Furthermore, 6 species were Bacillus-like new record species in China, including B. glycinifermentans(FJAT-47935, FJAT-47944, and FJAT-47907), B. hisashii(FJAT-47926), Bhargavaea cecembensis(FJAT-47948), Lysinibacillus acetophenoni(FJAT-47912, FJAT-47934, and FJAT-47919), Ornithinibacillus bavariensis(FJAT-47915), and Ornithinibacillus scapharcae(FJAT-47951). There were significant differences in the Bacillus-like species population size and structure during different composting and processing stages, with a size variation range from-80% to +100% and a species variety rate from 62% to 84%. The content of the Bacilluslike species populations was 1.14×106, 2.37×106 cfu·g-1 and 3.76×105 cfu·g-1 at the early(1 d), middle(30 d), and later(60 d)stages, respectively, of composting fermentation, and that was 4.3×105 cfu·g-1 after the production process. The indigenous Bacillus-like species had less influence on composting fermentation of the sugar-refinery filtering muds; the introduced Bacillus-like species during composting fermentation were mainly brought by the sugar-refinery filtering muds themselves. Therefore, the self-brought Bacillus-like species could fully develop their actions during the sugar-refinery filtering mud composting fermentation, although any additional Bacillus-like inoculation agent was not used. In conclusion, the Bacillus-like species are rich and their population size and structure are significantly different in the process of sugar-refinery filtering mud composting and processing.
The aim of this study is to determine dynamic changes in Bacillus-like species population in the process of sugar-refinery filtering mud composting fermentation to produce bio-organic fertilizers. The Bacillus-like species in the samples at different composting andprocessing stages were isolated and identified to analyze the dynamic changes in their population size and structure during the process, and to reveal their relationship with the indigenous Bacillus-like species. The results indicated that the Bacillus-like species were enriched in the process of sugar-refinery filtering mud composting fermentation. A total of 64 strains, belonging to 27 species within 7 genera, were isolated from 6 samples at different composting and processing stages. Among them, the genus Bacillus, including 16 species, was the absolutely dominant population. Furthermore, 6 species were Bacillus-like new record species in China, including B. glycinifermentans(FJAT-47935, FJAT-47944, and FJAT-47907), B. hisashii(FJAT-47926), Bhargavaea cecembensis(FJAT-47948), Lysinibacillus acetophenoni(FJAT-47912, FJAT-47934, and FJAT-47919), Ornithinibacillus bavariensis(FJAT-47915), and Ornithinibacillus scapharcae(FJAT-47951). There were significant differences in the Bacillus-like species population size and structure during different composting and processing stages, with a size variation range from-80% to +100% and a species variety rate from 62% to 84%. The content of the Bacilluslike species populations was 1.14×106, 2.37×106 cfu·g-1 and 3.76×105 cfu·g-1 at the early(1 d), middle(30 d), and later(60 d)stages, respectively, of composting fermentation, and that was 4.3×105 cfu·g-1 after the production process. The indigenous Bacillus-like species had less influence on composting fermentation of the sugar-refinery filtering muds; the introduced Bacillus-like species during composting fermentation were mainly brought by the sugar-refinery filtering muds themselves. Therefore, the self-brought Bacillus-like species could fully develop their actions during the sugar-refinery filtering mud composting fermentation, although any additional Bacillus-like inoculation agent was not used. In conclusion, the Bacillus-like species are rich and their population size and structure are significantly different in the process of sugar-refinery filtering mud composting and processing.
引文
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[2]谭宏伟,周柳强,谢如林,等.蔗糖生产中的有机废弃物资源化利用研究[J].大众科技,2016,18(5):110-112.TAN Hong-wei,ZHOU Liu-qiang,XIE Ru-lin,et al.Application of organic discarded goods in cane sugar production[J].Popular Science and Technology,2016,18(5):110-112.
[3]Kumar R,Verma D,Singh B L,et al.Composting of sugar-cane waste by-products through treatment with microorganisms and subsequent vermicomposting[J].Bioresource Technology,2010,101(17):6707-6711.
[4]Pérez C,Falero A,Hung B R,et al.Bioconversion of phytosterols to androstanes by mycobacteria growing on sugar cane mud[J].Journal of Industrial Microbiology&Biotechnology,2005,32(3):83-86.
[5]Rocky-Salimi K,Hashemi M,Safari M,et al.Valorisation of untreated cane molasses for enhanced phytase production by Bacillus subtilis K46b and its potential role in dephytinisation[J].Journal of the Science of Food and Agriculture,2017,97(1):222-229.
[6]Veana F,Martínez-Hernández J L,Aguilar C N,et al.Utilization of molasses and sugar cane bagasse for production of fungal invertase in solid state fermentation using Aspergillus niger GH1[J].Brazilian Journal of Microbiology,2014,45(2):373-377.
[7]Poopathi S,Mani C,Rajeswari G.Potential of sugarcane bagasse(agroindustrial waste)for the production of Bacillus thuringiensis israelensis[J].Tropical Biomedicine,2013,30(3):504-515.
[8]Techapun C,Poosaran N,Watanabe M,et al.Optimization of aeration and agitation rates to improve cellulase-free xylanase production by thermotolerant Streptomyces sp.Ab106 and repeated fed-batch cultivation using agricultural waste[J].Journal of Bioscience and Bioengineering,2003,95(3):298-301.
[9]谭宏伟.糖厂滤泥发酵制成生物有机肥[J].南方农业学报,2017,48(3):428-432.TAN Hong-wei.Bio-organic fertilizer made through fermentation of filter mud from sugar refinery[J].Journal of Southern Agriculture,2017,48(3):428-432.
[10]左见军,印华亮,王威威,等.甘蔗糖厂滤泥的资源化利用分析[J].企业科技与发展,2014(14):19-21.ZUO Jian-jun,YIN Hua-liang,WANG Wei-wei,et al.The comprehensive utilization of byproducts and wastes in cane sugar industry[J].Enterprise Science and Technology&Development,2014(14):19-21.
[11]刘晓举,陈宝娣,郭继强.利用糖厂滤泥生产根瘤菌菌剂[J].大连工业大学学报,2013,32(3):187-190.LIU Xiao-ju,CHEN Bao-di,GUO Ji-qiang.Production of rhizobia inoculant using sugar mill filter mud[J].Journal of Dalian Polytechnic University,2013,32(3):187-190.
[12]黄飒,欧宗喜.碳酸法糖厂滤泥在甘蔗种植应用上的探索[J].广西蔗糖,2012(2):20-25.HUANG Sa,OU Zong-xi.Application of carbonate sugar mill filter on sugar cane planting[J].Guangxi Sugarcane&Canesugar,2012(2):20-25.
[13]虎玉森,王兴民.糖厂滤泥中肥效成分的研究[J].甘肃农业大学学报,1998(1):87-89.HU Yu-sen,WANG Xing-min.Study on fertilizer effective components in filter mud of sugar mill[J].Journal of Gansu Agricultural University,1998(1):87-89.
[14]Wang X,Selvam A,Lau S S S,et al.Influence of lime and struvite on microbial community succession and odour emission during food waste composting[J].Bioresource Technology,2017,18(247):652-659.
[15]Zhong X Z,Ma S C,Wang S P,et al.A comparative study of composting the solid fraction of dairy manure with or without bulking material:Performance and microbial community dynamics[J].Bioresource Technology,2017,21(247):443-452.
[16]Zhao X,He X,Xi B,et al.Response of humic-reducing microorganisms to the redox properties of humic substance during composting[J].Waste Manag,2017,16(9):56-68.
[17]王守红,朱凌宇,徐荣,等.菌剂添加对牛粪堆肥氮素变化及腐熟度影响[J].安徽农业大学学报,2017,44(4):1-5.WANG Shou-hong,ZHU Ling-yu,XU Rong,et al.Nitrogen changes and maturity of cow manure composting influenced by microbial inoculum[J].Journal of Anhui Agricultural University.2017,44(4):1-5.
[18]赵利超,余从田,黄晓风,等.奶牛粪酸臭成分降解过程中微生物群落变化的研究[J].微生物学报,2017,57(6):1-9.ZHAO Li-chao,YU Cong-tian,HUANG Xiao-feng,et al.Study on microbial community changes during the degradation of dairy manure acid components[J].Acta Microbiologica Sinica,2017,57(6):1-9.
[19]徐大兵,佀国涵,徐祥玉,等.辅以多黏类芽孢杆菌堆肥提取液工艺及其对土壤微生物群落结构的影响[J].湖北农业科学,2017,56(4):634-639.XU Da-bing,LüGuo-han,XU Xiang-yu,et al.The technology research of pig manure compost extracts combined paenibacillus polymyxa and its influence on soil microbial community structure[J].Hubei Agricultural Sciences,2017,56(4):634-639.
[20]李敬波.牛粪堆肥过程中优势菌群变化及复合菌剂制备[D].大连:大连理工大学,2014.LI Jing-bo.Changes of dominant microbiota during cattle manure composting and preparation of a microbial inoculant for composting[D].Dalian:Dalian University of Technology,2014.
[21]肖勇.基于分子生物学技术的堆肥微生物群落、功能与应用研究[D].长沙:湖南大学,2011.XIAO Yong.Researches on the communities,functions and applications of microorganisms in compost based on molecular biological techniques[D].Changsha:Hunan University,2011.
[22]Logan N A,Berge O,Bishop A H,et al.Proposed minimal standards for describing new taxa of aerobic,endospore-forming bacteria[J].International Journal of Systematic and Evolutionary Microbiology,2009,59:2114-2121.
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