猪尸体好氧堆肥生物强化技术研究

发布时间：2018-03-11 19:13

  本文选题：猪尸体　切入点：复合菌剂　出处：《华中农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：好氧发酵堆肥简单实用、经济环保并能为农作物提供高效肥料,已经成为动物尸体无害化处理的重要方式,但同时存在发酵周期长、堆肥产品质量不高等缺陷。为了提高堆肥产品质量、缩短堆肥时间,本试验从猪尸体自然堆肥样品中分离出高效微生物,配制成复合菌剂,应用于猪尸体堆肥中,研究微生物菌剂的强化对猪尸体堆肥体系的影响。主要研究结果如下:(1)采集猪尸体自然堆肥样品,分离筛选高效微生物。共获得了35种微生物,其中24株细菌、9株真菌,2株放线菌。(2)通过滴定法和平板法测定35种菌株的脂肪酶活性和蛋白酶活性、淀粉酶活性,筛选出4株酶活较高的微生物;生理生化及16S rDNA鉴定这4株菌分别为:地衣芽孢杆菌、枯草芽孢杆菌、甲基营养型芽孢杆菌、解淀粉芽孢杆菌。(3)运用L9(34)正交实验的方法研究菌株不同添加量对混合发酵液中微生物增殖的影响。结果显示地衣芽孢杆菌、解淀粉芽孢杆菌、枯草芽孢杆菌、甲基营养型芽孢杆菌中,地衣芽孢杆菌的添加量是最主要因素。最优处理为处理A3B2C1D3(地衣芽孢杆菌0.75%,甲基营养型芽孢杆菌0.5%,解淀粉芽孢杆菌0.25%,枯草芽孢杆菌0.75%)时,活菌数可达到6.89×109cfu/m L,为试验所需组合,用于后续堆肥试验。(4)对猪尸体好氧堆肥体系施加复合微生物菌剂,菌剂强化组最高温度为58℃,50℃以上持续15d,而对照组最高温度为52℃,50℃以上持续10d;菌剂强化组和对照组堆体含水率的变化趋势相同,均从55%左右降至33%左右;菌剂强化组和对照组的总氮含量各为1.9%和2.7%;堆肥结束菌剂强化组发芽指数达到97.3%,而对照组发芽指数仅为40.5%。(5)利用16S rRNA高通量测序技术分析了猪尸体堆肥体系微生物群落的结构和动态变化。菌剂强化组第1个月细菌的丰富度chao1指数和多样性Shannon指数为1059和7.06,对照组为682和5.96,表明添加微生物菌剂可以增加堆肥起始微生物的丰富度和多样性指数。两组猪尸体堆肥过程中细菌的多样性相似,主要的优势菌群是Proteobacteria、Firmicutes和Actinobacteria,但是不同的时间其组成不同。(6)将堆肥环境参数和细菌群落数据进行典范对应分析(CCA),结果表明环境因子中,温度(p=0.002)、pH(p=0.004)、湿度(p=0.020)、全氮(p=0.064)对微生物群落丰度及其时空分布有显著影响。其中,温度与高温阶段S1(试验组)及类芽胞杆菌属最相关,GI(发芽指数)和腐熟阶段S6及假单胞菌属密切相关,全氮与腐熟阶段样D6(对照组)及石油降解菌属密切相关。因此,可以在猪尸体堆肥中添加类芽胞杆菌属、假单胞菌属、石油降解菌属菌,提高堆体的温度,加快有机质降解,增加氮含量,提高腐熟度。
[Abstract]:Aerobic fermentation compost is simple and practical, economical and environmental protection, and can provide high efficiency fertilizer for crops. It has become an important way of harmless treatment of animal carcasses, but it also has a long fermentation period. In order to improve the quality of composting products and shorten the composting time, high efficient microbes were isolated from the natural compost samples of pig carcasses, and the compound microbes were prepared and used in pig carcass composting. The main results are as follows: (1) the natural compost samples of pig carcasses were collected, and the high efficient microbes were isolated and screened. A total of 35 kinds of microbes were obtained. The lipase activity, protease activity and amylase activity of 35 strains were determined by titration and plate method, and 4 microbes with higher enzyme activity were screened out. Physiological, biochemical and 16s rDNA identified the four strains as Bacillus licheniformis, Bacillus subtilis and Bacillus subtilis. The effects of different dosage of Bacillus amylolyticus on the microorganism proliferation in mixed fermentation broth were studied by L9 / 34) orthogonal experiment. The results showed that Bacillus licheniformis, Bacillus amylolyticus, Bacillus subtilis, Bacillus subtilis, Bacillus licheniformis and Bacillus subtilis. The addition amount of Bacillus licheniformis is the most important factor in the methyl-nutritive Bacillus, and the best treatment is to deal with A3B2C1D3 (Bacillus licheniformis 0.7575, Bacillus methylnutrition 0.5, Bacillus amylolyticus 0.25g, Bacillus subtilis 0.75g). The number of live bacteria can reach 6.89 脳 10 9 cfur / m L, which is used in the subsequent composting test. 4) to apply the compound microbial agent to the aerobic composting system of pig carcass. The maximum temperature of the bacteria fortified group was over 58 鈩，

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