农业有机废物与城市生活垃圾堆肥高温期微生物种群结构比较

通过PCR、克隆文库方法分析了农业有机废物和城市垃圾堆肥高温期间细菌和真菌种群的多样性.提取堆肥高温期的DNA,PCR扩增,构建各高温期的16S rDNA和18S rDNA克隆文库,结果表明:农业有机废物和城市生活垃圾16S rDNA克隆文库中分别共有18个、21个OTUs,分别属于细菌域的14个、15个不同属,其18S rDNA克隆文库中分别共有8个、9个OTUs,分别属于细菌域的8个、9个不同属,推断农业有机废物堆体的优势菌为Bacillus megaterium、Rhizobium sp.、Phanerochaete chrysosporium、Penicillium sp.同属或同种的菌株;城市生活垃圾堆体的优势菌为Bacillus megaterium、Azospirillum sp.、Phanerochaete chrysosporium同种或同属的菌株.     

高通量测序技术分析猪粪堆肥过程中微生物群落结构变化

为了解猪粪堆肥过程中微生物群落结构组成及多样性的变化,采集猪粪堆肥过程的三个代表性样品—新鲜猪粪、高温堆肥、腐熟堆肥,利用Illumina Miseq高通量测序技术对16S rRNA V4～V5可变区序列进行测序,分别获得37 009、42 470、36 713条有效序列及328、280、160个操作分类单元(OTU)。Alpha多样性分析表明,在堆肥过程中微生物群落丰富度呈现降低趋势,而多样性呈现先上升后下降趋势。随着堆肥的进行,在门水平上,厚壁菌门、拟杆菌门和软壁菌门相对丰度降低,而变形菌门和放线菌门相对丰度升高;在属水平上,Turicibacter、Terrisporobacter、Parabacteroides、Clostridium sensu stricto、Corynebacterium等来自动物肠道的微生物相对丰度明显下降,Thermopolyspora、Thermomonospora、Thermobifida、Halocella等耐热耐盐微生物成为最主要优势菌。堆肥过程不同菌群优势度的变化是微生物与堆肥中各理化因子相互作用的结果。     

外接堆肥微生物在餐厨废弃物好氧堆肥中的应用

微生物是堆肥过程的主要驱动力,其群落在堆肥过程中不断演替,进而影响好氧发酵进程和堆肥的品质。大量研究表明：接种外源微生物可促进堆肥进程,加快有机物分解和堆肥腐熟,消解抗生素等有害物质等作用。餐厨废弃物油脂、盐、水含量高和易酸化等问题会影响好氧发酵过程中微生物的活动。通过特定样品的富集驯化、选择性培养,并结合先进的分子生物学技术,筛选具有不同降解功能的菌株,并以此为基础构建的微生物菌剂能够一定程度上克服餐厨废弃物用于好氧堆肥的限制性问题。主要阐述了餐厨废弃物好氧发酵过程中微生物菌群的演替规律,构建微生物菌剂的菌种类型及功能和不同菌剂对好氧发酵的作用及潜在影响机制,以期能够为相关微生物强化技术的研发提供一些参考。     

鸡粪堆肥的微生物变化规律研究

目的：研究鸡粪堆肥中细菌、放线菌和真菌数量的变化规律,为有机堆肥提供技术参数。方法：采取稀释倒平板法和涂布法对堆肥样品进行计数。结果：在发酵过程中,菌数量变化在肥料表面上基本是呈先下降后上升最后达到稳定状态。其中细菌在30cm和60cm深处趋于平稳,真菌在30cm和60cm深处先上升再下降再上升,在第12d后有个急速下降的过程。放线菌在30cm和60cm深处数量比较少。而在后熟发酵过程中,3种菌数的变化规律比较平稳。结论：细菌和真菌的生长和繁殖都会抑制放线茵的生长,其中细菌作用大,真菌次之。     

鸡粪堆肥的微生物变化规律研究

目的:研究鸡粪堆肥中细菌、放线菌和真菌数量的变化规律,为有机堆肥提供技术参数。方法:采取稀释倒平板法和涂布法对堆肥样品进行计数。结果:在发酵过程中,菌数量变化在肥料表面上基本是呈先下降后上升最后达到稳定状态。其中细菌在30cm和60cm深处趋于平稳,真菌在30cm和60cm深处先上升再下降再上升,在第12d后有个急速下降的过程。放线菌在30cm和60cm深处数量比较少。而在后熟发酵过程中,3种菌数的变化规律比较平稳结论:细菌和真菌的生长和繁殖都会抑制放线菌的生长,其中细菌作用大,真菌次之。     

变性梯度凝胶电泳在堆肥微生物研究中的应用

对当前堆肥中微生物种群分布及其对有机物分解作用的研究进行分析,论述了分子生物技术中的变性梯度凝胶电泳(DGGE)的特点。将DGCE与PCR扩增技术相结合,可用于研究自然菌种堆肥和人工培养驯化菌种堆肥过程中微生物的演替规律,为研究和筛选堆肥中的微生物提供更加有效、快速的信息,促进堆肥技术的发展。     

微生物燃料电池在固体废物堆肥中的应用进展

微生物燃料电池(Microbial fuel cell,MFC)是一种近几年快速发展的废物处理与能源化技术,可以与污水处理、污染物降解、脱盐等环境技术结合。微生物燃料电池与堆肥技术结合可以在处理日益增长的固体废弃物的同时回收能量,具有很好的发展前景。文中分析了堆肥微生物燃料电池系统的微生物特征,探讨了堆肥过程中影响微生物燃料电池产电性能的因素,包括电极,隔膜,供氧和构型。最后归纳说明了堆肥微生物电池作为一种新的废弃物处理技术的特点:较高的微生物量并可产生较高的电流密度;对不同环境的适应性强;可以自身调节温度,能源利用效率高;质子从阳极向阴极的移动会受到不同堆肥原料的影响。     

土霉素对堆肥过程中酶活性和微生物群落代谢的影响

摘 要;以猪粪和秸秆为试验材料,研究了土霉素对堆肥温度、种子发芽指数、C/N、纤维素酶、脲酶、过氧化氢酶以及微生物群落代谢的影响。结果表明, 0 mg/kg (对照处理)的堆肥在第 2 天上升到 50 ℃以上, 维持了 5 d,达到了无害化处理的要求。35 mg/kg 土霉素处理(A1 处理）第 4 天升到 51.0 ℃,其高温期维持了 1 d。70 mg/kg 土霉素处理(A2 处理)第 3 天升到 50 ℃以上,高温期维持了 2 d。105 mg/kg 土霉素处理(A3 处理)和 140 mg/kg 土霉素处理(A4 处理)的温度在整个堆肥期间均未达到 50 ℃。在堆肥结束时各个处理的种子发芽指数均达到 80 % 以上。CK（Control check, 对照处理）、A1、A2、A3 和 A4 处理的 C/N 由 34.50 分别降为 16.64、16.07、19.48、18.45 和 19.83。堆肥的第 1 天,A1、A2、A3 和 A4 处理对纤维素酶活性的抑制率分别为 60.30%、21.30%、48.81% 和 76.05%,第 3 天,土霉素对纤维素酶活性起促进作用,在 4—30 d,A3、A4 处理对纤维素酶活性有抑制作用。在堆肥的前期（1—3 d）,土霉素刺激脲酶活性,随着堆肥时间的延长,土霉素对脲酶活性由刺激变为抑制作用,在第 18—30 天 A3、A4 处理与 CK 相比有着显著的抑制作用。在堆肥第 1 天到第 18 天,土霉素基本上促进过氧化氢酶的活性,堆肥 18 d 之后土霉素对过氧化氢酶起着是抑制作用。用 Biolog (ECO Microplate)方法研究了土霉素对堆肥过程中微生物群落代谢的影响,结果表明,在升温期 CK 处理的 AWCD (Average Well Color Development, 平均颜色变化率)在培养 60 h 之后大于其他处理,高温期 A2 处理的 AWCD 值最高,在降温期 CK 的 AWCD 值一直是最高的。对 Shannon 指数进行分析显示,堆肥的初始阶段土霉素降低微生物群落的功能多样性,随着时间的延长,土霉素增加微生物群落的功能多样性。对微生物利用六大类碳源分析表明, 140 mg/kg 的土霉素浓度能够改变微生物利用碳源的种类。     

复合微生物菌剂在剩余污泥堆肥中的作用研究

随着我国社会经济的快速发展,社会经济活动中产生的污水和污泥处理问题也越来越严重,广泛存在于污水处理机构中的污泥成为一个日渐严重的问题,复合微生物菌剂在污水处理活动中剩余污泥的处理活动中,因为其对污泥无害化处理的积极作用而备受欢迎,本文将从复合微生物菌剂的角度出发,结合剩余污泥堆肥处理中的实际,对复合微生物菌剂在剩余污泥堆肥处理中的作用进行简要的分析。     

Microbial community succession and lignocellulose degradation during agricultural waste composting

The changes of microbial community during agricultural waste composting were successfully studied by quinone profiles. Mesophilic bacteria indicated by MK-7 and mesophilic fungi containing Q-9 as major quinone were predominant and seemed to be important during the initial stage of composting. Actinobacteria indicated by a series of partially saturated and long-chain menaquinones were preponderant during the thermophilic period. While Actinobacteria, fungi and some bacteria, especially those microbes containing MK-7(H4) found in Gram-positive bacteria with a low G+C content or Actinobacteria were found cooperate during the latter maturating period. Since lignocellulsoe is abundant in the agricultural wastes and its degradation is essential for the operation of composting, it’s important to establish the correlation between the quinone profiles changes and lignocellulose degradation. The microbes containing Q-9 or Q-10(H2) as major quinone were found to be the most important hemicellulose and cellulose degrading microorganisms during composting. While the microorganisms containing Q-9(H2) as major quinone and many thermophilic Actinobacteria were believed to be responsible for lignin degradation during agricultural waste composting.     

Microbial characterization during composting of municipal solid waste

This study investigates the prevailing physico-chemical conditions and microbial community; mesophilic bacteria, yeasts and filamentous fungi, bacterial spores, Salmonella and Shigella as well as faecal indicator bacteria: total coliforms, faecal coliforms and faecal Streptococci, present in a compost of municipal solid waste. Investigations were conducted in a semi-industrial pilot plant using a moderate aeration during the composting process. Our results showed that: (i) auto-sterilization induced by relatively high temperatures (60–55°C) caused a significant change in bacterial communities. For instance, Escherichia coli and faecal Streptococci populations decreased, respectively, from 2×10⁷ to 3.1×10³ and 10⁷ to 1.5×10³ cells/g waste dry weight (WDW); yeasts and filamentous fungi decreased from 4.5×10⁶ to 2.6×10³ cells/g WDW and mesophilic bacteria were reduced from 5.8×10⁹ to 1.8×10⁷ bacteria/g WDW. On the other hand, the number of bacterial spores increased at the beginning of the composting process, but after the third week their number decreased notably; (ii) Salmonella disappeared completely from compost by the 25th day as soon as the temperature reached 60°C; and (iii) the bacterial population increased gradually during the cooling phase. While Staphylococci seemed to be the dominant bacteria during the mesophilic phase and at the beginning of the thermophilic phase, bacilli predominated during the remainder of the composting cycle. The appearance of gram-negative rods (opportunistic pathogens) during the cooling phase may represent a serious risk for the sanitary quality of the finished product intended for agronomic reuse. Compost sonication for about 3 min induced the inactivation of delicate bacteria, in particular gram-negatives. By contrast, gram-positive bacteria, especially micrococcus, spores of bacilli, and fungal propagules survived, and reached high concentrations in the compost.     

不同微生物菌剂处理对猪粪堆肥中氨挥发的影响

研究不同微生物复合菌剂及添加比例对猪粪与木屑混合(鲜重比为鲜猪粪∶木屑9∶1)堆肥过程中NH₃挥发的影响.结果表明,在堆肥过程中,NH₃挥发主要产生在堆肥前期15 d的升温和高温期,添加3‰的微生物复合菌剂1、2和3对猪粪堆肥中NH₃挥发都有一定的抑制作用,减轻氮素损失与堆肥恶臭,添加5‰复合菌剂1有显著抑制作用（P＜0.05）.     

Microbial communities involved in anaerobic degradation of alkanes

Saturated hydrocarbons are quantitatively the most abundant fraction among all petroleum hydrocarbons. Significant advances have been made in the understanding of the anaerobic biodegradability of alkanes in terms of the microorganisms involved and the biochemical pathways over the past two decades. They can be used as carbon and energy sources by diverse physiological groups of microorganisms (isolates or consortia) grown under chlorate-reducing, nitrate-reducing, sufidogenic or methanogenic conditions. Two general biochemical mechanisms have been proposed for the initial activation of alkanes including addition of fumarate and carboxylation. However, glycyl radical enzymes dependent fumarate addition which yields alkyl-substituted succinate appear to be the most commonly shared mechanism for the anaerobic attack of alkanes under various redox conditions by phylogenetically diverse microorganisms. The genes encoding the candidate alkylsuccinate synthase have been recently described in alkane-degrading sulfate- and nitrate-reducers as well as in hydrocarbon-rich environments. Alternative mechanisms may also be available depending on the alkane-degrading microbial community and electron acceptors utilized.     

Response of compost maturity and microbial community composition to pentachlorophenol (PCP)-contaminated soil during composting

Two composting piles were prepared by adding to a mixture of rice straw, vegetables and bran: (i) raw soil free from pentachlorophenol (PCP) contamination (pile A) and (ii) PCP-contaminated soil (pile B). It was shown by the results that compost maturity characterized by water soluble carbon (WSC), TOC/TN ratio, germination index (GI) and dehydrogenase activity (DA) was significantly affected by PCP exposure, which resulted in an inferior degree of maturity for pile B. DGGE analysis revealed an inhibited effect of PCP on compost microbial abundance. The bacteria community shifts were mainly consistent with composting factors such as temperature, pH, moisture content and substrates. By contrast, the fungal communities were more sensitive to PCP contamination due to the significant correlation between fungal community shifts and PCP removal. Therefore, the different microbial community compositions for properly evaluating the degree of maturity and PCP contamination were suggested.     

Investigation of the microbial community structure and activity as indicators of compost stability and composting process evolution

In a bid to identify suitable microbial indicators of compost stability, the process evolution during windrow composting of poultry manure (PM), green waste (GW) and biowaste was studied. Treatments were monitored with regard to abiotic factors, respiration activity (determined using the SOUR test) and functional microflora. The composting process went through typical changes in temperature, moisture content and microbial properties, despite the inherent feedstock differences. Nitrobacter and pathogen indicators varied as a monotonous function of processing time. Some microbial groups have shown a potential to serve as fingerprints of the different process stages, but still they should be examined in context with respirometric tests and abiotic parameters. Respiration activity reflected well the process stage, verifying the value of respirometric tests to access compost stability. SOUR values below 1 mg O₂/g VS/h were achieved for the PM and the GW compost.     

Changes in biochemical and microbiological parameters during the period of rapid composting of dairy manure with rice chaff

Various parameters were measured during the period of composting of dairy manure and rice chaff in different ratios (dairy manure/rice chaff=V/V, pile 1: 75/25; pile 2: 80/20; pile 3: 85/15) to evaluate their suitability as indicators for the composting process. The temperature in pile 1 increased rapidly and remained above 60 °C for 30 days, while the temperature in pile 3 increased slowly relative to the other two piles. Furthermore, the degradation of organic substrates, as indicated by the reduction of C/N ratio, was rapid in pile 1 (below 20% 28 days after beginning of the composting). The major fluctuations of various water-soluble fractions in all piles were observed during the first 3 weeks, and the results in general showed that the highest microbial populations and enzymatic activities also appeared in this phase. Various parameters indicated that the rapid composting method was a feasible one for treating agricultural wastes.     

Microbial community structure changes during bioremediation of PAHs in an aged coal-tar contaminated soil by in-vessel composting

The microbial community structure changes of an aged-coal-tar soil contaminated with polycyclic aromatic hydrocarbons (PAHs) were investigated during simulated bioremediation at the laboratory-scale using an in-vessel composting approach. The composting reactors were operated using a logistic three-factor factorial design with three temperatures (T=38, 55 or 70 °C), four soil to green-waste amendment ratios (S:GW=0.6:1, 0.7:1, 0.8:1 or 0.9:1 on a dry weight basis) and three moisture contents (MC=40%, 60% or 80%). Relative changes in microbial populations were investigated by following the dynamics of phospholipid fatty acid (PLFA) signatures using a ¹³C-labeled palmitic acid internal standard and sensitive GC/MS analysis during in-vessel composting over 98 days. The results of this investigation indicated that fungal to bacterial PLFA ratios were significantly influenced by temperature (p<0.05), and Gram-positive to Gram-negative bacterial ratios were significantly influenced by temperature (p<0.001) and S:GW ratio (p<0.01) during in-vessel composting. Additionally, the Gram-positive to Gram-negative bacterial ratios were correlated to the extent of PAH losses (p<0.005) at 70 °C.