【摘要】：堆肥是處理牛糞等農(nóng)牧業(yè)固體廢棄物的主要方式,自然堆肥存在著腐熟時(shí)間長、營養(yǎng)損耗大和二次污染較大等問題。在堆肥中添加外源菌劑,增強(qiáng)微生物數(shù)量分解有機(jī)質(zhì)活性、減少氮素?fù)p失、加速腐熟進(jìn)程、提高堆肥產(chǎn)品質(zhì)量,已得到廣泛證實(shí)。微生物是驅(qū)動(dòng)堆肥中氮素轉(zhuǎn)化的主要因素,與堆肥的進(jìn)程和堆肥質(zhì)量密切相關(guān),然而,關(guān)于堆肥中,尤其是添加外源菌劑后堆肥中氮素轉(zhuǎn)化微生物的群落變化的研究鮮見報(bào)道。本文研究了添加菌劑對(duì)牛糞稻草堆肥理化性質(zhì)的影響,并采用PCR-DGGE技術(shù)研究了菌劑對(duì)堆肥中氨化細(xì)菌、氨氧化古菌和氨氧化細(xì)菌的動(dòng)態(tài)及多樣性影響,旨在加深對(duì)堆肥中氮素轉(zhuǎn)化知識(shí)的理解并為堆肥生產(chǎn)實(shí)踐提供理論指導(dǎo)。本文主要研究結(jié)果如下:1.添加菌劑縮短堆肥周期,促進(jìn)堆肥腐熟。添加菌劑在3~36d提高堆肥溫度(P0.05),最高溫度達(dá)到62℃;在3~45d提高氮素含量并降低含水率(P0.05);在21~45d減少總碳的消耗(P0.01);在3、7、21和36d能夠降低牛糞堆肥的碳氮比(P0.05),比值更接近腐熟標(biāo)準(zhǔn);在21~36d增加牛糞堆肥的銨態(tài)氮含量(P0.05);在7~36d能夠有效地增加牛糞堆肥的硝態(tài)氮含量(P0.05)。2.通過對(duì)氨化細(xì)菌的測定分析發(fā)現(xiàn),添加菌劑在3d、15d和21d顯著增加氨化細(xì)菌數(shù)量;并提高氨化細(xì)菌多樣性指數(shù),在堆肥3d多樣性指數(shù)最大;自然堆肥中出現(xiàn)的特異氨化細(xì)菌為Bacillus horikoshii strain CMB14。自然堆肥和添加菌劑堆肥中均存在氨化細(xì)菌類型包括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,這些菌株均屬于嗜堿芽孢桿菌(Bacillus halodurans strain)、蘇云金芽孢桿菌(Bacillus thuringiensis strain)、枯草芽孢桿菌(Bacillus subtitles strain)、煙酸芽孢桿菌(Bacillus niacin strain)、環(huán)狀芽孢桿菌(Bacillus circulars)、嗜熱芽孢桿菌(Bacillus horikoshii strain)。3.通過對(duì)氨氧化古菌的測定分析發(fā)現(xiàn),添加菌劑堆肥中氨氧化古菌多樣性指數(shù)增加,在堆肥36d多樣性指數(shù)最大。在自然堆肥第15d中出現(xiàn)的特異氨氧化古菌為Uncultured ammonia-oxidizing archaeon clone XFNAOA18 amo A,在菌劑堆肥7d和15d發(fā)現(xiàn)特異性氨氧化古菌為Uncultured archaeon clone SEPHIV_2 amo A。自然堆肥和添加菌劑均菌種類型包括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,這些菌株均為不可培養(yǎng)類型,屬于中溫泉古菌奇古菌(thaumarchaeote)、泉古菌(crenarchaeote)。4.通過對(duì)氨氧化細(xì)菌的測定分析發(fā)現(xiàn),添加菌劑提高氨氧化細(xì)菌多樣性指數(shù)。在堆肥15d多樣性指數(shù)最大,在自然堆肥堆肥第3d出現(xiàn)特異性氨氧化細(xì)菌為Nitrosospira sp.Wyke8(amo A)gene。自然堆肥和添加菌劑堆肥中均氨氧化細(xì)菌類型包括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,這些菌株屬于亞硝化螺菌屬(Nitrosospira sp.)、亞硝化球菌屬(Nitrosococcus sp.)及不可培養(yǎng)的變形菌(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.
【學(xué)位授予單位】：東北農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S141.4

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本文編號(hào)：2222402