Methanogenic diversity and activity in municipal solid waste landfill leachates

Archaeal microbial communities present in municipal solid waste landfill leachates were characterized using a 16S rDNA approach. Phylogenetic affiliations of 239 partial length 16S rDNA sequences were determined. Sequences belonging to the order Methanosarcinales were dominant in the clone library and 65% of the clones belonged to the strictly acetoclastic methanogenic family Methanosaetaceae. Sequences affiliated to the metabolically versatile family Methanosarcinaceae represented 18% of the retrieved sequences. Members of the hydrogenotrophic order Methanomicrobiales were also recovered in limited numbers, especially sequences affiliated to the genera Methanoculleus and Methanofollis. Eleven euryarchaeal and thirteen crenarchaeal sequences (i.e. 10%) were distantly related to any hitherto cultivated microorganisms, showing that archaeal diversity within the investigated samples was limited. Lab-scale incubations were performed with leachates mixed with several methanogenic precursors (acetate, hydrogen, formate, methanol, methylamine). Microbial populations were followed using group specific 16S rRNA targeted fluorescent oligonucleotidic probes. During the incubations with acetate, acetoclastic methanogenesis was rapidly induced and led to the dominance of archaea hybridizing with probe MS1414 which indicates their affiliation to the family Methanosarcinaceae. Hydrogen and formate addition induced an important acetate synthesis resulting from the onset of homoacetogenic metabolism. In these incubations, species belonging to the family Methanosarcinaceae (hybridizing with probe MS1414) and the order Methanomicrobiales (hybridizing with probe EURY496) were dominant. Homoacetogenesis was also recorded for incubations with methanol and methylamines. In the methanol experiment, acetoclastic methanogenesis took place and archaea hybridizing with probe MS821 (specific for Methanosarcina spp.) were observed to be the dominant population. These results confirm that acetoclastic methanogenesis performed by the members of the order Methanosarcinales is predominant over the hydrogenotrophic and methylotrophic pathways in landfill leachates.     

垃圾填埋场氧化亚氮排放控制研究进展

填埋是国内外城市生活垃圾处理的一种主要方式.垃圾填埋场是温室气体氧化亚氮(N2O)和甲烷(CH4)的重要排放源.作为一种高效痕量的温室气体,N2O具有极高的潜在增温效应,其每分子潜在的增温作用是二氧化碳(CO2)的296倍.而且N2O能在大气中长期稳定存在,对臭氧层具有较强的破坏作用.本文针对垃圾填埋场N2O排放的控制研究,概述了垃圾填埋处理过程中主要排放源的N2O排放及其影响因素,提出了现阶段适应我国垃圾填埋场N2O排放控制的一系列措施,并展望了垃圾填埋场温室气体N2O排放控制理论和技术的研究方向.     

Archaeal community compositions at different depths (up to 30 m) of a municipal solid waste landfill in Taiwan as revealed by 16S rDNA cloning analyses

Archaeal 16S rDNA clone libraries were constructed for samples taken at 10, 20 and 30 m depth in a landfill, which corresponded approximately 3, 6 and 9 years operation, respectively. Sequencing and phylogenetic analyses of representative clones showed that all of the rDNAs were closely related to typical methanogens. The distributions of phylotypes in clone libraries were similar to each other. Dominant clones in all the clone libraries were closely related to thermophilic species, such as Methanothermobacter thermautotrophicus, suggesting that the temperatures at these sites were high. This was supported by the results of H₂-dependent methanogenic activity tests, showing that the activities of all samples at 55 °C were much higher than those at 25 °C.     

Molecular phylogenetic diversity of bacteria associated with the leachate of a closed municipal solid waste landfill

A 16S rDNA-based molecular study was performed to determine the nature of the bacterial constituents of the leachate from a closed municipal solid waste landfill. Total community DNA was extracted and bacterial 16S rRNA genes were subsequently amplified and cloned. Recombinant rDNA clones in the library were randomly selected, and they were sequenced for a single run and then grouped. A total of 76 sequence types representing 138 randomly selected nonchimeric clones were identified. Full-length sequencing and phylogenetic analysis of the sequence types revealed that more than 90% of the screened clones were affiliated with low-G+C gram-positive bacteria (38.4%), Proteobacteria (35.5%), the Cytophaga Flexibacter Bacteroides group (11.6%), and Spirochaetes (5.1%). Minor portions were affiliated with Verrucomicrobia (2.9%), candidate division OP11 (2.2%), and the green nonsulfur bacteria, Cyanobacteria and the Deinococcus Thermus group (each <1.0%). Although some rDNA sequences clustered with genera or taxa that were classically identified within anaerobic treatment systems and expected with known functions, a substantial fraction of the clone sequences showed relatively low levels of similarity with any other reported rDNA sequences and thus were derived from unknown taxa. These results suggest that bacterial communities in landfill environment are far more complex than previously expected and remain largely unexplored.     

Archaeal diversity and community development in deep-sea hydrothermal vents

Over the past 35 years, researchers have explored deep-sea hydrothermal vent environments around the globe and studied a number of archaea, their unique metabolic and physiological properties, and their vast phylogenetic diversity. Although the pace of discovery of new archaeal taxa, phylotypes and phenotypes in deep-sea hydrothermal vents has slowed recently, bioinformatics and interdisciplinary geochemistry-microbiology approaches are providing new information on the diversity and community composition of archaea living in deep-sea vents. Recent investigations have revealed that archaea could have originated and dispersed from ancestral communities endemic to hydrothermal vents into other biomes on Earth, and the community structure and productivity of chemolithotrophic archaea are controlled primarily by variations in the geochemical composition of hydrothermal fluids.     

Bioreactor landfill technology in municipal solid waste treatment: An overview

In recent years, due to an advance in knowledge of landfill behaviour and decomposition processes of municipal solid waste, there has been a strong thrust to upgrade existing landfill technologies for optimizing these degradation processes and thereafter harness a maximum of the useful bioavailable matter in the form of higher landfill gas generation rates. Operating landfills as bioreactors for enhancing the stabilization of wastes is one such technology option that has been recently investigated and has already been in use in many countries. A few full-scale implementations of this novel technology are gaining momentum in landfill research and development activities. The publication of bioreactor landfill research has resulted in a wide pool of knowledge and useful engineering data. This review covers leachate recirculation and stabilization, nitrogen transformation and corresponding extensive laboratory- and pilot-scale research, the bioreactor landfill concept, the benefits to be derived from this bioreactor landfill technology, and the design and operational issues and research trends that form the basis of applied landfill research.     

The microbial ecology of anaerobic cellulose degradation in municipal waste landfill sites: evidence of a role for fibrobacters

Cellulose is reputedly the most abundant organic polymer in the biosphere, yet despite the fundamental role of cellulolytic microorganisms in global carbon cycling and as potential sources of novel enzymes for biotechnology, their identity and ecology is not well established. Cellulose is a major component of landfill waste and its degradation is therefore a key feature of the anaerobic microbial decomposition process. Here, we targeted a number of taxa containing known cellulolytic anaerobes (members of the bacterial genus Fibrobacter, lineages of Clostridium clusters I, III, IV and XIV, and anaerobic fungi of the Neocallimastigales) in landfill leachate and colonized cellulose 'baits' via PCR and quantitative PCR (qPCR). Fibrobacter spp. and Clostridium clusters III, IV and XIV were detected in almost all leachate samples and cluster III and XIV clostridia were the most abundant (1-6% and 1-17% of total bacterial 16S rRNA gene copies respectively). Two landfill leachate microcosms were constructed to specifically assess those microbial communities that colonize and degrade cellulose substrates in situ. Scanning electron microscopy (SEM) of colonized cotton revealed extensive cellulose degradation in one microcosm, and Fibrobacter spp. and Clostridium cluster III represented 29% and 17%, respectively, of total bacterial 16S rRNA gene copies in the biofilm. Visible cellulose degradation was not observed in the second microcosm, and this correlated with negligible relative abundances of Clostridium cluster III and Fibrobacter spp. (≤ 0.1%), providing the first evidence that the novel fibrobacters recently detected in landfill sites and other non-gut environments colonize and degrade cellulose substrates in situ.     

Telomere shortening in women resident close to waste landfill sites

Several studies demonstrate links between environmental stress and index of reduced health, including risk factors for cardiovascular disease, reduced immune function and cancer risks. We investigated the hypothesis that pollution, as an environmental stress, impacts health by modulating the rate of cellular aging in healthy pregnant women. Our research looked at the effects that illegal waste sites have on the localized population of pregnant women in Campania, Italy. As is often the case in illegal dumping, the effects on the population are often seen well before knowing what specific agents in the soil and water are responsible. Here we provide evidence that the pollution in this region is significantly associated with higher oxidative stress, shorter telomere length and lower telomerase activity, which are known determinants of cell senescence and aging-related meiotic dysfunction in women, in peripheral blood mononuclear cells from healthy pregnant women, subjected to therapeutic abortion in the second trimester of pregnancy. These findings may have implications for understanding how, at the cellular level, environmental stress may promote earlier onset of age-related diseases.     

准好氧填埋渗滤液水质变化特性研究

在大型模拟填埋试验装置(21 m×3.8 m×6.0 m)上,研究了准好氧填埋渗滤液水质的主要指标COD_Cr、BOD、NH₃⁺-N和pH的变化特性.结果表明,准好氧填埋结构下渗滤液COD_Cr、BOD浓度下降很快,没有出现在传统填埋场累积的现象,并且封场后39周分别降为173和30 mg·L^-1;NH₃⁺-N浓度下降更为显著,第39周降为1 mg·L^-1,下降率达到99.6%,为渗滤液后续处理解决了NH₃⁺-N浓度过高的难题;pH值在前2周略低于7,第3周后一直呈弱碱性.根据实验数据,拟合了准好氧填埋结构渗滤液污染物的衰减方程.     

Mutagenic activity of the leachate of municipal solid waste landfill.

Organic concentrates were recovered using XAD-2/8 resin adsorption from the leachates of municipal solid waste landfills and their mutagenic activities were tested for 8 months using the Ames Salmonella/microsome assay. Highly polluted leachates (COD and BOD > or = 40 mg/l) generally had equal or higher mutagenic activities than lightly polluted leachates (COD and BOD < 40 mg/l). But there was no clear difference in mutagenicity per amount of concentrate between the two leachates. These results suggest that the mutagenic activity of landfill leachate is decided to some degree by the organic concentration in the leachate. The mutagenic activities detected even in lightly polluted leachates were not so low as those of various kind of surface waters ever reported. It is suggested that it is important to investigate the mutagenic activity of the leachate for evaluation of the impact of landfill leachate on the environment.     

Molecular biological detection and characterization of Clostridium populations in municipal landfill sites

Primer sets specific for 16S rRNA genes were designed for four phylogenetic groups of clostridia known to contain mesophilic cellulolytic species. Specific amplification of these groups from landfill leachate DNA extracts demonstrated the widespread occurrence of clostridia from the Clostridium thermocellum and C. leptum groups. In contrast, the C. botulinum group was never detected, and the C. coccoides-C. lentocellum group was only occasionally detected. Amplification products were analyzed by temporal thermal gel electrophoresis to generate profiles of the clostridial groups and to identify dominant bands. Sequence analysis of 17 landfill clones confirmed that the primers were specific for the clostridial subgroups and that the cloned sequences had a close relationship with known cellulose-degrading clostridia. The primers have therefore been authenticated for use in the rapid identification of clostridia in anaerobic environments.     

Bacterial community diversity assessment in municipal solid waste compost amended soil using DGGE and ARISA fingerprinting methods

Bacterial community structure and diversity of Tunisian agricultural soil treated with different amounts of municipal solid waste compost (MSWC) and other fertilizers were studied using DGGE and ARISA fingerprinting methods. Sequence analysis of dominant DGGE bands revealed the presence of three major clusters, Cytophaga/Flexibacter/Bacteroides (CFB) group, Proteobacteria and Acidobacteria group. Using ARISA profiles, dominant populations were assigned to low and high GC Gram positive bacteria, Cyanobacteria, Spirochetes and Cytophagales. The two methods revealed the absence of significant bacterial community shifts related to the different MSWC applications. Moreover, indigenous bacterial population of the used loam-clayey soil was observed to limit proliferation and survival of Proteobacteria, initially dominant in MSWC and farmyard manure. Effectiveness of the two methods for soil bacterial community studying was shown. While DGGE was more accurate for bacterial identification, ARISA was more practical for handling and rapid estimation of dominant bacteria.     

Anaerobic digestion of municipal solid waste as a treatment prior to landfill

Anaerobic digestion of organic fraction of municipal solid waste was conducted in pilot-scale reactor based on high-solid combined anaerobic digestion process. This study was performed in two runs. In Run 1 and Run 2, pre-stage flushing and micro-aeration were conducted to determine their effect in terms of enhancing hydrolysis and acidification in ambient condition. In Run 2, after pre-stage, the methane phase (methanogenesis) was started-up after pH adjustment and inoculum addition in mesophilic condition. Acidified leachate produced in pre-stage was used for percolation during active methane phase. At the end of methane phase, air flushing was conducted before unloading the digesters. Hydrolysis and acidification yield of 140 g C/kg TS and 180 g VFA/kg TS were achieved, respectively in pre-stage. Micro-aeration exhibited an equivocal result in terms of enhancing hydrolysis/acidification; however it showed a positive effect in methane phase performance and this needed further investigation. Leachate percolation during methane phase showed an enhanced methanization when compared to the reactors without leachate percolation. After 60 days, 260 l CH(4)/kg VS was obtained. Based on the waste methane potential, 75% biogas conversion and 61% VS degradation were achieved.     

Quantification and diversity of the archaeal community in a landfill site

At a sea-based, solid waste disposal site, methanogenic organisms were quantified by molecular approaches. The samples collected for analysis were from anaerobic leachate of the landfill site. When the DNA extracted from the leachate was examined by a quantitative PCR method using domain-specific 16S rDNA primers, archaeal DNA represented 2-3% of the total extracted DNA. On the basis of cloning and sequence comparison of the archaeal PCR products, more than half of the sequences belonged to Euryarchaeota, particularly relatives of the genus Methanosaeta. The cloning analysis suggested that the majority of methane emitted from the landfill site originated from the acetate-utilizing Methanosaeta.     

Archaeal diversity in Icelandic hot springs

Whole-cell density gradient extractions from three solfataras (pH 2.5) ranging in temperature from 81 to 90 degrees C and one neutral hot spring (81 degrees C, pH 7) from the thermal active area of Hveragerethi (Iceland) were analysed for genetic diversity and local geographical variation of Archaea by analysis of amplified 16S rRNA genes. In addition to the three solfataras and the neutral hot spring, 10 soil samples in transects of the soil adjacent to the solfataras were analysed using terminal restriction fragment length polymorphism (t-RFLP). The sequence data from the clone libraries in combination with 14 t-RFLP profiles revealed a high abundance of clones clustering together with sequences from the nonthermophilic I.1b group of Crenarchaeota. The archaeal diversity in one solfatara was high; 26 different RFLP patterns were found using double digestion of the PCR products with restriction enzymes AluI and BsuRI. The sequenced clones from this solfatara belonged to Sulfolobales, Thermoproteales or were most closest related to sequences from uncultured Archaea. Sequences related to group I.1b were not found in the neutral hot spring or the hyperthermophilic solfatara (90 degrees C).     

Comparative life cycle assessment of two landfill technologies for the treatment of municipal solid waste

Goal and Scope  The potential environmental impacts associated with two landfill technologies for the treatment of municipal solid waste (MSW), the engineered landfill and the bioreactor landfill, were assessed using the life cycle assessment (LCA) tool. The system boundaries were expanded to include an external energy production function since the landfill gas collected from the bioreactor landfill can be energetically valorized into either electricity or heat; the functional unit was then defined as the stabilization of 600 000 tonnes of MSW and the production of 2.56x10⁸ MJ of electricity and 7.81x10⁸ MJ of heat. Methods  Only the life cycle stages that presented differences between the two compared options were considered in the study. The four life cycle stages considered in the study cover the landfill cell construction, the daily and closure operations, the leachate and landfill gas associated emissions and the external energy production. The temporal boundary corresponded to the stabilization of the waste and was represented by the time to produce 95% of the calculated landfill gas volume. The potential impacts were evaluated using the EDIP97 method, stopping after the characterization step. Results and Discussion  The inventory phase of the LCA showed that the engineered landfill uses 26% more natural resources and generates 81% more solid wastes throughout its life cycle than the bioreactor landfill. The evaluated impacts, essentially associated with the external energy production and the landfill gas related emissions, are on average 91% higher for the engineered landfill, since for this option 1) no energy is recovered from the landfill gas and 2) more landfill gas is released untreated after the end of the post-closure monitoring period. The valorization of the landfill gas to electricity or heat showed similar environmental profiles (1% more raw materials and 7% more solid waste for the heat option but 13% more impacts for the electricity option). Conclusion and Recommendations  The methodological choices made during this study, e.g. simplification of the systems by the exclusion of the identical life cycle stages, limit the use of the results to the comparison of the two considered options. The validity of this comparison could however be improved if the systems were placed in the larger context of municipal solid waste management and include activities such as recycling, composting and incineration.     

Mediating N2O emissions from municipal solid waste landfills: Impacts of landfill operating conditions on community structure of ammonia-oxidizing bacteria in cover soils

Chemolithotrophic ammonia-oxidizing bacteria (AOB) can produce N₂O, a highly potent greenhouse gas. Denaturing gradient gel electrophoresis (DGGE) analyses of the ammonia monooxygenase structural gene (amoA) and 16S rDNA gene were used to investigate the AOB community structure in the cover soils of municipal solid waste (MSW) landfills under three operating conditions: (a) MSW with soil cover, (b) MSW with soil cover, irrigation piping and vegetation, and (c) MSW covered with high-density polyethylene (HDPE) liner, soil cover, irrigation piping and vegetation. AOB species in MSW cover soils were significantly distinguished by the operation of HDPE liner isolation. The community structures of the Nitrosomonas europaea-like AOB species dominated in soils without HDPE liner isolation, whether vegetation and irrigation with landfill leachate existed or not, whereas Nitrospira-like AOB species dominated in soils with HDPE liner isolation. Lower N₂O flux from the soils with HDPE liner isolation would be partially related to these special community structures.     

生活垃圾堆肥基质草坪建植体系接种草坪土壤线虫多样性特征

通过城市生活垃圾堆肥基质草坪建植体系接种草坪土壤线虫,研究了线虫数量、科属分类、营养类群等线虫多样性变化.结果表明,接种草坪土壤线虫 4 个月后,垃圾堆肥基质中共鉴定出线虫4科7属,包括植物寄生性线虫、食真菌线虫和食细菌线虫3个营养类群;垃圾堆肥基质草坪建植体系中线虫总数明显少于土壤基质.在接种草坪线虫的土壤基质草坪建植体系中,植物寄生线虫为优势营养类群,盘旋线虫属仍为优势属,相对多度达到83.3%,显著高于本底土壤45.1%.在接种草坪土壤线虫的垃圾堆肥基质草坪建植体系中,土壤本底中处于优势属的植物寄生类线虫如盘旋线虫属和螺旋线虫属基本消失了,表明垃圾堆肥基质对植物寄生类群的生长与繁殖表现出明显的抑制作用,优势属也发生了很大变化,食细菌类群的头叶属和丽突属成为优势属,相对多度分别为67.0%和14.0%.因此,从草坪土壤线虫危害角度来说,以垃圾堆肥作为草坪基质将会为草坪植物生长创造良好的基质环境.