菌剂对牛粪堆肥及其氮素转化微生物的影响
发布时间：2018-09-04 14:18
【摘要】：堆肥是处理牛粪等农牧业固体废弃物的主要方式,自然堆肥存在着腐熟时间长、营养损耗大和二次污染较大等问题。在堆肥中添加外源菌剂,增强微生物数量分解有机质活性、减少氮素损失、加速腐熟进程、提高堆肥产品质量,已得到广泛证实。微生物是驱动堆肥中氮素转化的主要因素,与堆肥的进程和堆肥质量密切相关,然而,关于堆肥中,尤其是添加外源菌剂后堆肥中氮素转化微生物的群落变化的研究鲜见报道。本文研究了添加菌剂对牛粪稻草堆肥理化性质的影响,并采用PCR-DGGE技术研究了菌剂对堆肥中氨化细菌、氨氧化古菌和氨氧化细菌的动态及多样性影响,旨在加深对堆肥中氮素转化知识的理解并为堆肥生产实践提供理论指导。本文主要研究结果如下:1.添加菌剂缩短堆肥周期,促进堆肥腐熟。添加菌剂在3~36d提高堆肥温度(P0.05),最高温度达到62℃;在3~45d提高氮素含量并降低含水率(P0.05);在21~45d减少总碳的消耗(P0.01);在3、7、21和36d能够降低牛粪堆肥的碳氮比(P0.05),比值更接近腐熟标准;在21~36d增加牛粪堆肥的铵态氮含量(P0.05);在7~36d能够有效地增加牛粪堆肥的硝态氮含量(P0.05)。2.通过对氨化细菌的测定分析发现,添加菌剂在3d、15d和21d显著增加氨化细菌数量;并提高氨化细菌多样性指数,在堆肥3d多样性指数最大;自然堆肥中出现的特异氨化细菌为Bacillus horikoshii strain CMB14。自然堆肥和添加菌剂堆肥中均存在氨化细菌类型包括Bacillus thuringiensis strain DL10、Bacillus subtilis strain g6l、Bacillus nealsonii strain WS1、Bacillus sp.VITJSS、Bacillus halodurans strain BG5、Bacillus niacini strain TN17、Bacillus circulans、Bacillus subtilis strain VB9,这些菌株均属于嗜碱芽孢杆菌(Bacillus halodurans strain)、苏云金芽孢杆菌(Bacillus thuringiensis strain)、枯草芽孢杆菌(Bacillus subtitles strain)、烟酸芽孢杆菌(Bacillus niacin strain)、环状芽孢杆菌(Bacillus circulars)、嗜热芽孢杆菌(Bacillus horikoshii strain)。3.通过对氨氧化古菌的测定分析发现,添加菌剂堆肥中氨氧化古菌多样性指数增加,在堆肥36d多样性指数最大。在自然堆肥第15d中出现的特异氨氧化古菌为Uncultured ammonia-oxidizing archaeon clone XFNAOA18 amo A,在菌剂堆肥7d和15d发现特异性氨氧化古菌为Uncultured archaeon clone SEPHIV_2 amo A。自然堆肥和添加菌剂均菌种类型包括Uncultured archaeon clone 2-39 amo A、Uncultured archaeon clone 5-68 amo A、Uncultured archaeon clone A0619 amo A、Uncultured ammonia-oxidizing archaeon clone QTWAOA32 amo A、Uncultured archaeon clone A2816 amo A、Uncultured archaeon clone A6702 amo A、Uncultured thaumarchaeote clone OTU_3 amo A、Uncultured crenarchaeote clone PB75-13 amo A、Uncultured archaeon clone POTU4 amo A,这些菌株均为不可培养类型,属于中温泉古菌奇古菌(thaumarchaeote)、泉古菌(crenarchaeote)。4.通过对氨氧化细菌的测定分析发现,添加菌剂提高氨氧化细菌多样性指数。在堆肥15d多样性指数最大,在自然堆肥堆肥第3d出现特异性氨氧化细菌为Nitrosospira sp.Wyke8(amo A)gene。自然堆肥和添加菌剂堆肥中均氨氧化细菌类型包括Nitrosococcus oceani strain AFC24P(amo A)gene、Uncultured bacterium gp1a amo A gene、Uncultured bacterium gp18 amo A gene、Nitrosospira sp.En I299(amo A)gene、Nitrosospira sp.En13(amo A)gene、Nitrosococcus oceani strain SW N.oceani(amo A)gene、Nitrosococcus oceani strain AFC36(amo A)gene、Uncultured proteobacterium isolate DGGE gel band L-3(amo A)gene、Uncultured proteobacterium isolate DGGE gel band L-2(amo A)gene、Uncultured Nitrosospira sp.clone ISA00172(amo A)gene、Uncultured Nitrosospira sp.clone ISA00164(amo A)gene、Uncultured Nitrosospira sp.clone ISA00160(amo A)gene,这些菌株属于亚硝化螺菌属(Nitrosospira sp.)、亚硝化球菌属(Nitrosococcus sp.)及不可培养的变形菌(Uncultured proteobacterium)。
[Abstract]:Composting is the main way to treat solid wastes of agriculture and animal husbandry, such as cow dung. Natural composting has many problems, such as long maturity time, large nutrient loss and secondary pollution. Adding exogenous microbial agents to compost can enhance the activity of microbial quantitative decomposition of organic matter, reduce nitrogen loss, accelerate the process of decomposition, and improve the quality of compost products. It is confirmed that microorganism is the main factor driving nitrogen transformation in compost, which is closely related to composting process and composting quality. However, there are few reports on the community changes of nitrogen-transforming microorganisms in compost, especially when adding exogenous microbial agents. The effects of microbial agents on the dynamics and diversity of ammoniated bacteria, ammonia-oxidizing archaea and ammonia-oxidizing bacteria in compost were studied by PCR-DGGE technique in order to deepen the understanding of nitrogen transformation knowledge in compost and provide theoretical guidance for compost production practice. The addition of microbial agents increased the composting temperature (P 0.05) at 3-36 days, and the highest temperature reached 62; increased nitrogen content and decreased water content (P 0.05) at 3-45 days; decreased total carbon consumption (P 0.01) at 21-45 days; decreased the C/N ratio (P 0.05) at 3, 7, 21 and 36 days; increased ammonium nitrogen content (P 0.05) at 21-36 days. Nitrate nitrogen content in cattle manure compost could be effectively increased in 7-36 days (P Hoikoshii strain CMB14. Ammoniated bacteria were found in both natural compost and microbial agent compost, including Bacillus thuringiensis strain DL10, Bacillus subtilis strain g6l, Bacillus nealsonii strain WS1, Bacillus sp. VITJSS, Bacillus halodurans strain BG5, Bacillus cinniacini strain TN17, Bacillus circulans, Bacillus subtilis LIS strain VB9, these strains belong to Bacillus halodurans strain, Bacillus thuringiensis strain, Bacillus subtitles strain, Bacillus niacin strain, Bacillus circulars, Bacillus horikos Hii strain). 3. The diversity index of ammonia-oxidizing archaea in compost with microbial agents increased, and the diversity index of ammonia-oxidizing archaea in compost with microbial agents was the highest at 36 days. The specific ammonia-oxidizing archaeon XFNAOA18 amo A in 15 days of natural compost was Uncultured ammonia-oxidizing archaeon clone XFNAOA18 amo A, which was found at 7 days and 15 days of compost with microbial agents. Specific ammonia-oxidizing archaea were Uncultured archaeon clone SEPHIV_2 amo A. Natural composting and addition of microbial agents included Uncultured archaeon clone 2-39 amo A, Uncultured archaeon clone 5-68 amo A, Uncultured archaeon clone A0619 amo A, Uncultured ammonia-oxidizing archaeon clone QTWA32 amo A, Uncultured a. A rchaeon clone A2816 amo A, Uncultured archaeon clone A6702 amo A, Uncultured thaumarchaeote clone OTU_3 amo A, Uncultured crenarchaeote clone PB75-13 amo A, Uncultured archaeon clone POTU4 amo A. These strains are all non-culturable and belong to the middle-hot spring archaea, crenarchaeote and crenarchaeote. 4. Ammonia-oxidizing bacteria increased the diversity index of ammonia-oxidizing bacteria by adding microbial agents. The diversity index of ammonia-oxidizing bacteria was the highest on the 15th day of composting, and the specific ammonia-oxidizing bacteria was Nitrosospira sp. Wyke8 (amo A) gene on the 3rd day of natural composting. CCUs oceanI strain AFC24P (amo A) gene, Uncultured bacterium gp1a amo A gene, Uncultured bacterium gp18 amo A gene, Nitrosospira sp.En I299 (amo A) gene, Nitrosospira sp.En13 (amo A) gene, Nitrosococcus oceanI strain SW N.oceani (amo A) gene, Nitrosococco oceanius strain AFC36 (amo A) gene, Unculted proteobacteriu strain AFC36 (amo A) gene M isolate DGGE gel band L-3 (amo A) gene, Uncultured proteobacterium isolate DGGE gel band L-2 (amo A) gene, Uncultured Nitrosospira sp. clone ISA00172 (amo A) gene, Uncultured Nitrosospira sp. clone ISA00164 (amo A) gene, Uncultured Nitrospira sp. clone ISA00160 (amo A) gene, Uncultured Nitrospira sp. Nitrosospira sp., Nitrosococcus sp., and Uncultured Proteobacterium.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S141.4
本文编号：2222402
[Abstract]:Composting is the main way to treat solid wastes of agriculture and animal husbandry, such as cow dung. Natural composting has many problems, such as long maturity time, large nutrient loss and secondary pollution. Adding exogenous microbial agents to compost can enhance the activity of microbial quantitative decomposition of organic matter, reduce nitrogen loss, accelerate the process of decomposition, and improve the quality of compost products. It is confirmed that microorganism is the main factor driving nitrogen transformation in compost, which is closely related to composting process and composting quality. However, there are few reports on the community changes of nitrogen-transforming microorganisms in compost, especially when adding exogenous microbial agents. The effects of microbial agents on the dynamics and diversity of ammoniated bacteria, ammonia-oxidizing archaea and ammonia-oxidizing bacteria in compost were studied by PCR-DGGE technique in order to deepen the understanding of nitrogen transformation knowledge in compost and provide theoretical guidance for compost production practice. The addition of microbial agents increased the composting temperature (P 0.05) at 3-36 days, and the highest temperature reached 62; increased nitrogen content and decreased water content (P 0.05) at 3-45 days; decreased total carbon consumption (P 0.01) at 21-45 days; decreased the C/N ratio (P 0.05) at 3, 7, 21 and 36 days; increased ammonium nitrogen content (P 0.05) at 21-36 days. Nitrate nitrogen content in cattle manure compost could be effectively increased in 7-36 days (P Hoikoshii strain CMB14. Ammoniated bacteria were found in both natural compost and microbial agent compost, including Bacillus thuringiensis strain DL10, Bacillus subtilis strain g6l, Bacillus nealsonii strain WS1, Bacillus sp. VITJSS, Bacillus halodurans strain BG5, Bacillus cinniacini strain TN17, Bacillus circulans, Bacillus subtilis LIS strain VB9, these strains belong to Bacillus halodurans strain, Bacillus thuringiensis strain, Bacillus subtitles strain, Bacillus niacin strain, Bacillus circulars, Bacillus horikos Hii strain). 3. The diversity index of ammonia-oxidizing archaea in compost with microbial agents increased, and the diversity index of ammonia-oxidizing archaea in compost with microbial agents was the highest at 36 days. The specific ammonia-oxidizing archaeon XFNAOA18 amo A in 15 days of natural compost was Uncultured ammonia-oxidizing archaeon clone XFNAOA18 amo A, which was found at 7 days and 15 days of compost with microbial agents. Specific ammonia-oxidizing archaea were Uncultured archaeon clone SEPHIV_2 amo A. Natural composting and addition of microbial agents included Uncultured archaeon clone 2-39 amo A, Uncultured archaeon clone 5-68 amo A, Uncultured archaeon clone A0619 amo A, Uncultured ammonia-oxidizing archaeon clone QTWA32 amo A, Uncultured a. A rchaeon clone A2816 amo A, Uncultured archaeon clone A6702 amo A, Uncultured thaumarchaeote clone OTU_3 amo A, Uncultured crenarchaeote clone PB75-13 amo A, Uncultured archaeon clone POTU4 amo A. These strains are all non-culturable and belong to the middle-hot spring archaea, crenarchaeote and crenarchaeote. 4. Ammonia-oxidizing bacteria increased the diversity index of ammonia-oxidizing bacteria by adding microbial agents. The diversity index of ammonia-oxidizing bacteria was the highest on the 15th day of composting, and the specific ammonia-oxidizing bacteria was Nitrosospira sp. Wyke8 (amo A) gene on the 3rd day of natural composting. CCUs oceanI strain AFC24P (amo A) gene, Uncultured bacterium gp1a amo A gene, Uncultured bacterium gp18 amo A gene, Nitrosospira sp.En I299 (amo A) gene, Nitrosospira sp.En13 (amo A) gene, Nitrosococcus oceanI strain SW N.oceani (amo A) gene, Nitrosococco oceanius strain AFC36 (amo A) gene, Unculted proteobacteriu strain AFC36 (amo A) gene M isolate DGGE gel band L-3 (amo A) gene, Uncultured proteobacterium isolate DGGE gel band L-2 (amo A) gene, Uncultured Nitrosospira sp. clone ISA00172 (amo A) gene, Uncultured Nitrosospira sp. clone ISA00164 (amo A) gene, Uncultured Nitrospira sp. clone ISA00160 (amo A) gene, Uncultured Nitrospira sp. Nitrosospira sp., Nitrosococcus sp., and Uncultured Proteobacterium.
【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S141.4
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