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.     

Nitrogen amendment of green waste impacts microbial community,enzyme secretion and potential for lignocellulose decomposition

Microorganisms involved in biomass deconstruction are an important resource for organic waste recycling and enzymes for lignocellulose bioconversion. The goals of this study were to examine the impact of nitrogen amendment on microbial community restructuring, secretion of xylanases and endoglucanases, and potential for biomass deconstruction. Communities were cultivated aerobically at 55 °C on green waste (GW) amended with varying levels of NH₄Cl. Bacterial and fungal communities were determined using 16S rRNA and ITS region gene sequencing and PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) was applied to predict relative abundance of genes involved in lignocellulose hydrolysis. Nitrogen amendment significantly increased secretion of xylanases and endoglucanases, and microbial activity; enzyme activities and cumulative respiration were greatest when nitrogen level in GW was between 4.13–4.56 wt% (g/g), but decreased with higher nitrogen levels. The microbial community shifted to one with increasing potential to decompose complex polymers as nitrogen increased with peak potential occurring between 3.79–4.45 wt% (g/g) nitrogen amendment. The results will aid in informing the management of nitrogen level to foster microbial communities capable of secreting enzymes that hydrolyze recalcitrant polymers in lignocellulose and yield rapid decomposition of green waste.     

Chemical and structural evolution of humic acids during organic waste composting

The effects of the composting process on the chemical and structural properties of humic acids have been studied in seven different organic waste mixtures from different origin. Only slight changes in elemental composition have been found in the humic acids after the composting process pointing to a more aromatic structure with higher proportions of oxygen and nitrogen. Functional groups were the most sensitive to the changes caused by the composting process showing a marked increase in the total acidity and phenolic, carboxyl and carbonyl groups. Gel permeation chromatography showed a slight increase in the average molecular weight of the humic acids. Infrared spectroscopy did not show important differences in humic acid structure but a clear decrease in the intensity of the bands in the region 3000–2850 cm^-1 corresponding to the aliphatic fractions. As a general result, the composting process yields humic acids in which the elemental and functional composition are closer to that of the more humified soil humic acids.     

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。     

Novel static composting method for bioremediation of olive mill waste

This paper presents results obtained on the evaluation of static composting process aimed at bioremediation of the hazardous solid olive mill waste (OMW). The static composting process carried out in gas-permeable polyethylene bags followed the fluctuating temperature and oxygen profiles similar to those seen in aerated composting systems. Static composting resulted in apparent increases and decreases in values for total nitrogen and C:N ratios respectively during the process. The amount of nitrogen (>3%) in the composting end product was in agreement with the Italian legislation (Decreto Legislativo 29 aprile 2010, n. 75) specification for nitrogen fertilizer. A gradual decrease in polyphenols during the storage of compost resulted in a non-phytotoxic composted organic matter high in humic substances. Different respirometric tests also stated high biological stability of the end compost product.     

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。     

微生物在有机固废堆肥中的作用与应用

好氧堆肥是实现有机固体废弃物资源化利用的主流处理方式.堆肥腐熟是一个由微生物主导的生理生化过程,堆料通过微生物发酵实现矿质化、腐殖化和无害化,转变成腐熟的有机肥.传统的好氧堆肥存在发酵周期长、养分损失、恶臭及温室气体排放等不足.在堆肥过程中添加微生物是弥补传统好氧堆肥缺陷、提高堆肥品质和功效的有效方法.近年来,国内外在...     

Changes in the microbial community structure during thermophilic composting of manure as detected by the quinone profile method

Quinone profiles and physico-chemical properties were measured to characterize the microbial community structure during a 14-day thermophilic composting of cattle manure mixed with rice straw as a bulking agent. The change in total quinone content (TQ) and the divergence of quinone (DQ) indicated that the microbial biomass reached a peak followed by a decrease, whereas the microbial community diversity increased continuously during the composting process. The high mole fraction of ubiquinones in the raw materials, and at the beginning of the composting period suggested that fungi and/or Proteobacteria were present. The predominance of MK-7 from days 3 to 7 suggested that Bacillus spp. were the main microbial species. An increase in partially saturated and long-chain menaquinones during the latter composting period indicated that the proliferation of various species of Actinobacteria was occurring. The microbial community structure, as expressed by TQ and DQ, corresponded well to physico-chemical properties such as the C/N ratio, pH, O₂ consumption and compost mass reduction.     

Monitoring of microbial populations and their cellulolytic activities during the composting of municipal solid wastes

Seasonal changes in microbial populations and the activities of cellulolytic enzymes were investigated during the composting of municipal solid wastes at Damietta compost plant, Egypt. The changes in temperature, pH and carbon/nitrogen (C/N) ratio were also monitored. The results obtained showed that the temperatures of the windrows in all seasons reached the maximum after 3 weeks of composting and then decreased by the end of the composting period (35 days), but did not reach ambient temperature. Marked changes in pH values of the composts in all seasons were found, but generally, the pH was near neutrality. Significant increases in the size of the microbial populations were obtained in autumn and spring seasons compared to summer and winter seasons. The activities of cellulases were also higher in the autumn and spring seasons than in the summer and winter seasons. The decrease in C/N ratio in autumn and spring was higher than in summer and winter. It was evident that the degradation of organic matter increased by an increase in the microflora and its cellulolytic activities.     

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.     

超高温堆肥促进氮素减损的微生物机制研究进展

堆肥中氮的循环在很大程度上依赖微生物驱动的氮素转化。传统高温堆肥最高堆温普遍在55-60℃,温度的提高有利于缩短堆肥周期和提高堆肥品质。超高温堆肥作为近年来快速发展的新兴技术,不但能突破传统堆肥工艺堆温低的局限,并且持续的超高温调控了堆肥微生物组、堆肥环境与氮素的互作,减少了氮素的损失。本文综述了堆体的氮循环过程及超高温堆肥技术在保氮方面的显著优势,以及超高温堆肥过程中具有氮代谢功能的优势微生物种群及其影响因素,重点介绍有关超高温堆肥控制氮素损失的作用机制研究进展,同时对超高温堆肥现有研究中存在的问题进行分析并探讨解决途径。     

接种微生物菌剂对猪粪堆肥过程中细菌群落多样性的影响

利用PCR-DGGE方法研究了接种外源微生物菌剂对鲜猪粪高温好氧堆肥过程中细菌群落多样性的影响.结果表明：接种外源微生物菌剂可以促进堆肥的顺利进行,比不接种处理的高温期提前2 d.DGGE图谱分析表明,堆肥中优势细菌群落组成发生了明显的更迭现象,不同堆肥时期细菌群落的Shannon-Wiener指数呈显著差异.目的条带克隆测序结果表明,整个堆肥过程Clostridium stercorarium subsp. thermolacticum sp.一直是优势菌属,不经培养细菌、Bacillus coagulans sp.、Clostridium thermocellum sp.在接种外源微生物菌剂处理的第10、16天成为优势菌属,不经培养的Firmicutes sp.和不经培养的 delta proteobacterium分别在未接种外源微生物菌剂处理堆肥发酵的第5天和第16天成为优势菌属.非优势菌属Ureibacillus thermosphaericus、不经培养的Silvimonas sp.出现在堆肥腐熟后期,不经培养的土壤细菌主要出现在堆肥初期和高温初期.UPGMC聚类分析表明,接种外源微生物菌剂明显影响了堆肥不同时期的细菌群落结构组成.堆肥化过程中细菌DGGE图谱主成分分析表明,细菌群落变化主要受外源接种微生物菌剂的影响.     

微生物菌剂对猪粪堆肥中细菌群落结构的影响

以猪粪和小麦秸秆做堆肥试验,处理组添加外源微生物菌剂,利用常规方法对堆肥样品进行理化性状测定,采用高通量测序技术分析堆肥过程中细菌群落特征。理化性状测定结果表明: 添加外源菌剂可延长堆肥高温时间,降低堆肥发酵末期的pH,增加全氮含量,加快C/N的下降。主成分分析表明: 外源菌剂影响堆肥样品细菌群落的稳定性。门分类水平上,厚壁菌门、变形菌门和绿弯菌门的相对丰度在处理组中较高;纲分类水平上,梭状芽孢杆菌纲、α-变形菌纲和γ-变形菌纲在处理组的升温期和高温期相对丰度增加;科分类水平上,小单孢菌科和梭状芽孢杆菌纲的消化链球菌科、梭菌科以及盐厌氧菌科的相对丰度在处理组的升温期和高温期均呈上升趋势。Pearson相关性分析表明,盐胞菌属与外源菌剂呈显著正相关,而氨苄芽孢杆菌属与外源菌剂呈显著负相关。研究表明,猪粪堆肥中添加外源菌剂可使堆肥的理化性质和细菌群落结构均发生显著变化。     

葡萄籽对猪粪秸秆高温堆肥中微生物群落与碳氮含量的影响

以猪粪与秸秆(鲜质量10.5∶1)为基础,在自制的强制通风静态堆肥反应箱中进行堆肥化试验,研究添加8%葡萄籽对猪粪秸秆高温堆肥中微生物群落演替和碳氮转化的影响.在堆肥化的30 d里,分7次采集不同时期的堆肥样品,测定堆肥中微生物区系、微生物生理群的数量及堆肥碳氮含量.结果表明：添加葡萄籽使堆肥中细菌数量略高、放线菌数量显著增加、真菌数量明显降低,细菌/放线菌下降;氨化细菌和反硝化细菌数量降低;而硝化细菌、固氮菌和纤维素分解菌数量增多;铵态氮和有机碳含量下降,而硝态氮含量明显提高.堆肥中硝态氮含量与放线菌数量呈极显著正相关关系.添加葡萄籽使堆体升温快且高温期稳定,堆肥含水率波动较小,从而使堆肥高温期放线菌和亚硝化细菌的波动较小,数量较高,有利于堆肥中硝态氮含量的增加.     

鸡粪好氧堆肥过程中细菌群落结构和功能分析

为了明确鸡粪好氧堆肥过程中细菌群落结构和功能的变化,采用高通量测序技术测定了好氧堆肥前、中、后3个时期样品的16S rRNA基因序列,并进行了生物信息学分析。结果表明, 3个堆肥阶段中仅有10%左右的分类操作单元(operational taxonomic units, OTUs)具有阶段特异性;不同发酵阶段细菌α多样性指数ACE、Chao1和Simpson均呈现先升高后降低的趋势,但各阶段间差异不显著(P<0.05)。3个发酵阶段优势菌门类相同,但丰度存在差异。线性判别分析[line discriminant analysis (LDA) effect size, LEfSe]法对细菌生物标志分析表明,从门到属水平共有49个物种,堆肥前期样品组中显著富集的物种最多,后期最少。原核分类群功能注释(functional annotation of prokaryotic taxa,FAPROTAX)对细菌功能多样性分析表明,堆肥前期细菌功能多样性最高,而随着发酵进行细菌功能富集程度增加、多样性降低。该研究为畜禽粪污好氧堆肥过程调控提供理论支撑和技术指导。     

Continuous thermophilic composting (CTC) for rapid biodegradation and maturation of organic municipal solid waste

Fewer and fewer municipal solid wastes are treated by composting in China because of the disadvantages of enormous investment, long processing cycle and unstable products in a conventional composting treatment. In this study, a continuous thermophilic composting (CTC) method, only a thermophilic phase within the process, has been applied to four bench-scale composting runs, and further compared with a conventional composting run by assessing the indexes of pH, total organic carbon (TOC), total Kjeldahl nitrogen (TKN), C/N ratio, germination index (GI), specific oxygen uptake rate (SOUR), dissolved organic carbon (DOC) and dehydrogenase activity. After composting for 14 days, 16 days, 18 days and 19 days in the four CTC runs, respectively, mature compost products were obtained, with quality similar to or better than which had been stabilized for 28 days in run A. The products from the CTC runs also showed favorable stability in room temperature environment after the short-term composting at high temperature. The study suggested CTC as a novel method for rapid degradation and maturation of organic municipal solid wastes.     

Monitoring respiration and biological stability during sludge composting with a modified dynamic respirometer

A static electrolytic respirometer was adapted to work under dynamic conditions using an internal airflow recirculation system that passed air through sludge/straw solid mixtures. Airflow reduced oxygen transfer limitations and increased the maximum respiration rate and k_La values, indicating that the observed value may have been close to the actual biodegradation rate. Airflow caused sludge drying, so the sample moisture was controlled by air humidification. To apply the optimised respirometric technique, a pilotscale composting process was developed. Some commonly used respiration indices (RI₂₄, AT₄) were used to measure the final compost stability. RI₂₄ indices without airflow were underestimated during the thermophilic composting stage. Once the easily biodegradable carbon was consumed, the static and dynamic RI₂₄ indices were nearly identical. Because of the dynamic procedure, the respiration rate was likely controlled by the biochemical reaction and not by the mass transfer. The respiration indices indicated that the final compost was unstable.     

Development of compost maturity and Actinobacteria populations during full-scale composting of organic household waste

AIMS: This study investigates changes in microbiological and physicochemical parameters during large-scale, thermophilic composting of a single batch of municipal organic waste. The inter-relationships between the microbial biomass and community structure as well as several physicochemical parameters and estimates of maturation were evaluated. METHODS AND RESULTS: Analyses of signature fatty acids with the phospholipid fatty acid and ester-linked methods showed that the total microbial biomass was highest during the early thermophilic phase. The contribution of signature 10Me fatty acids from Actinobacteria indicated a relatively constant proportion around 10% of the microbial community. However, analyses of the Actinobacteria species composition with a PCR-denaturing gradient gel electrophoresis approach targeting 16S rRNA genes demonstrated clear shifts in the community structure. CONCLUSIONS: This study demonstrates that compost quality, particularly maturity, is linked to the composition of the microbial community structure, but further studies in other full-scale systems are needed to validate the generality of these findings. SIGNIFICANCE AND IMPACT OF THE STUDY: The combination of signature lipid and nucleic acid-based analyses greatly expands the specificity and the scope for assessing the microbial community composition in composts. The results presented in this study give new information on how the development of the compost microbial community is connected to curing and maturation in the later stages of composting, and emphasizes the role of Actinobacteria in this respect.     

Two-phase olive mill waste composting: community dynamics and functional role of the resident microbiota

In this study, physico-chemical modifications and community dynamics and functional role of the resident microbiota during composting of humid husk from a two-phase extraction system (TPOMW) were investigated. High mineralization and humification of carbon, low loss of nitrogen and complete degradation of polyphenols led to the waste biotransformation into a high-quality compost. Viable cell counts and denaturing gradient gel electrophoresis (DGGE) profiling of the 16S rRNA genes showed that the thermophilic phase was characterized by the strongest variations of cell number, the highest biodiversity and the most variable community profiles. The isolation of tannin-degrading bacteria (e.g. Lysinibacillus fusiformis, Kocuria palustris, Tetrathiobacter kashmirensis and Rhodococcus rhodochrous) suggested a role of this enzymatic activity during the process. Taken together, the results indicated that the composting process, particularly the thermophilic phase, was characterized by a rapid succession of specialized bacterial populations with key roles in the organic matter biotransformation.     

Temperature effect on Fusarium oxysporum f.sp. melonis survival during horticultural waste composting

AIMS: The aim of this work was to study the effect of high temperatures generated during composting process, on the phytopathogen fungus Fusarium oxysporum f.sp. melonis. This investigation was achieved by both in vivo (semipilot-scale composting of horticultural wastes) and in vitro (lab-scale thermal treatments) assays. METHODS AND RESULTS: Vegetable residues infected with F. oxysporum f.sp. melonis were included in compost piles. Studies were conducted in several compost windrows subjected to different treatments. Results showed an effective suppression of persistence and infective capacity, as this process caused complete fungal elimination after 2-3 days of composting. In order to confirm the effect of high temperature during this process, in vitro experiments were carried out. Temperature values of 45, 55 and 65 degrees C were tested. All three treatments caused the elimination of fungal persistence. Treatment at 65 degrees C was especially effective, whereas 45 degrees C eliminated fungal persistence only after 10 days. CONCLUSIONS: The composting process is an excellent alternative for the management of plant wastes after harvesting, as this procedure is able to suppress infective capacity of several harmful phytopathogens such as F. oxysporum f.sp. melonis. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium oxysporum f.sp. melonis is a plant pathogen fungus specially important in the province of Almería (south-east Spain), where intensive greenhouse horticulture is very extended. High temperatures reached during composting of horticultural plant wastes ensure the elimination of phytopathogen microorganisms such as F. oxysporum f.sp. melonis from vegetable material, providing an adequate hygienic quality in composts obtained.