A survey of ammonia-assimilating micro-organisms in cattle manure composting

AIMS: To evaluate the ammonia-assimilating abilities of micro-organisms isolated from cattle manure composting processes and to determine the distribution of cultivable species of ammonia-assimilating micro-organisms in microbial communities during the composting processes. METHODS AND RESULTS: Compost samples were collected from four stages of treatment. Trypto soya agar was used for the isolation of ammonia-assimilating aerobes. Many of the isolates showed high ammonia-assimilating ability in a medium containing basal components and a compost extract. Partial 16S ribosomal DNA sequencing showed that the cultivable species of highly efficient ammonia-assimilating isolates changed during the composting process. The community structure of micro-organisms and actinomycetes was analysed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Two species of actinomycetes identified by PCR-DGGE coincided with those found among the cultured isolates. CONCLUSIONS: Ammonia-assimilating micro-organisms obtained by the cultivation method were not predominant in the microbial community during the composting process: however certain cultured actinomycetes were members of predominant species in the actinomycetes community. SIGNIFICANCE AND IMPACT OF THE STUDY: Ammonia assimilation by micro-organisms is one of the important mechanisms for ammonia retention in the composting process. Cultivable actinomycetes are a means for preventing ammonia emission from the composting process.     

Comparison of organic matter degradation and microbial community during thermophilic composting of two different types of anaerobic sludge

Changes in organic matter degradation and microbial communities during thermophilic composting were compared using two different types of anaerobic sludge, one from mesophilic methane fermentation, containing a high concentration of proteins (S-sludge), and the other from thermophilic methane fermentation, containing high concentrations of lipids and fibers (K-sludge). The difference in the organic matter degradation rate corresponded to the difference in the organic matter constituents; the CO(2) evolution rate was greater in the composting of S-sludge than of K-sludge; moreover, the NH(3) evolution resulting from the protein degradation was especially higher in the composting of S-sludge. Then the differences in the microbial communities that contributed to each composting were determined by the PCR-DGGE method. Ureibacillus sp., which is known as a degrader with high organic matter degradation activity, was observed during the composting of S-sludge, whereas Thermobifida fusca, which is a well known thermophilic actinomycete that produces enzymes for lignocellulose degradation, were observed during the composting of K-sludge.     

Microbial succession during a composting process as evaluated by denaturing gradient gel electrophoresis analysis

Microbial succession during a laboratory-scale composting process of garbage was analysed by denaturing gradient gel electrophoresis (DGGE) combined with measurement of physicochemical parameters such as temperature, pH, organic acids, total dissolved organic carbon and water-soluble humic substance. From the temperature changes, a rapid increase from 25 to 58 degrees C and then a gradual decrease, four phases were recognized in the process as follows; mesophilic (S), thermophilic (T), cooling (C) and maturing (M). The polymerase chain reaction-amplified 16S rDNA fragments with universal (907R) and eubacterial (341F with GC clamp) primers were subjected to DGGE analysis. Consequently, the DGGE band pattern changed during the composting process. The direct sequences from DGGE bands were related to those of known genera in the DNA database. The microbial succession determined by DGGE was summarized as follows: in the S phase some fermenting bacteria, such as lactobacillus, were present with the existing organic acids; in the T phase thermophilic bacillus appeared and, after the C phase, bacterial populations were more complex than in previous phases and the phylogenetic positions of those populations were relatively distant from strains so far in the DNA database. Thus, the DGGE method is useful to reveal microbial succession during a composting process.     

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.     

有机物料种类及腐熟水平对土壤微生物群落的影响

应用Biolog方法研究了温室盆栽番茄条件下,施用不同种类及不同腐熟水平的有机物料对土壤微生物群落的影响,施用有机物料60d后取土分析土壤微生物群落多样性及土壤微生物对Biolog微平板中胺、氨基酸、糖、羧酸、聚合物和其它类碳源的利用情况,结果表明,施用有机物料可提高土壤微生物群落多样性,施用新鲜酒糟的多样性指数略高于施用腐熟10d酒糟,牛粪不同腐熟水平对多样性影响显著,且对多样性具有正向或负向的影响;对照和施用酒糟的土壤微生物对聚合物的利用率高于施用牛粪处理,施用新鲜物料处理的土壤微生物对聚合物的利用率高于施用腐熟物料处理。     

施氮对不同有机碳水平桉树林土壤微生物群落碳代谢的影响

土壤微生物群落碳代谢功能既受土壤氮素水平的影响,也与土壤有机碳水平密切相关,但二者如何共同影响土壤微生物群落碳代谢功能的研究尚不多见。以我国南方广泛种植的桉树林为对象,采用野外控制实验比较研究了4种施氮处理(对照:0kg/hm2,低氮:84.2 kg/hm2,中氮:166.8 kg/hm2,高氮:333.7 kg/hm2)对有机碳水平差异显著的两桉树林样地土壤微生物群落碳代谢功能的影响,结果表明:(1)两种有机碳水平桉树林土壤微生物群落碳代谢强度和代谢碳源丰富度显著不同,高有机碳水平桉树林土壤微生物群落碳代谢强度和代谢碳源丰富度显著高于低有机碳水平桉树林(P0.01);(2)施氮显著改变了桉树林土壤微生物群落的碳代谢强度和代谢碳源丰富度(P0.05),随着施氮水平的升高,土壤微生物群落碳代谢强度和代谢碳源丰富度均呈现先增加后降低的变化规律,但是高、低有机碳水平桉树林土壤微生物群落碳代谢强度和代谢碳源丰富度对施氮梯度的响应各不相同,高、低有机碳水平桉树林的土壤微生物群落碳代谢指标分别在中氮、低氮处理中达到最高值;(3)施氮影响土壤微生物群落代谢的碳源类型主要是碳水化合物类、氨基酸类和羧酸类,土壤微生物生物量是影响土壤微生物碳代谢强度和代谢碳源丰富度的重要因素。由此可知,施氮对土壤微生物碳代谢功能影响,也与土壤本底中有机碳水平的调节有关,所以在研究土壤微生物群落对施氮等条件的响应时,不能忽略土壤中有机碳水平。     

The stimulatory effects of surfactants on composting of waste rich in cellulose

The chemical surfactant Tween 80 and biosurfactant rhamnolipid were respectively added to the composting substrate, a mixture of rice straw and bran, and their effects on the composting process were investigated. Samples were analysed for microbial communities of bacteria, actinomycetes and fungi, carboxymethylcellulose hydrolysis (CMCase) and xylanase activities, cellulose and hemicellulose fractions, water-soluble carbon (WSC) contents in the substrates, organic matter contents and pH values during the composting process. The results showed that both Tween 80 and rhamnolipid had slight stimulatory effects on the microbial populations of bacteria, actinomycetes and fungi. In addition, rhamnolipid increased the peak xylanase activity 15% higher than that of the control, while Tween 80 increased the maximum CMCase activity 35% higher than that of the control. As a result of the increased enzyme activities, treatments with Tween 80 and rhamnolipid were of higher WSC contents than the control during the whole composting process. Accordingly, the composting process was accelerated by the surfactants, since the organic matter was decomposed more quickly and the breakdown of cellulose and hemicellulose was better in the treatments with Tween 80 and rhamnolipid.     

Changes in the chemical characteristics of water-extractable organic matter during composting and their influence on compost stability and maturity

Composting is the biochemical transformation of waste organic matter by microorganisms whose metabolism occurs in the water-soluble phase. Therefore, a study of the changes occurring in compost dissolved organic matter can be useful for assessing its stability and maturity. In light of the variety of parameters generally utilized to study composting processes, this work aims at identifying the major chemical processes that occur in solution and their influence on the attainment of stability and maturity with composting time. Compost stability, assessed by means of respirometric analysis which determined oxygen demand as a result of mineralization of the compost's organic matter, and compost maturity evaluated with Lepidium sativum L. seed bioassays, were found to be highly related to the nature and content of water-soluble organic matter. Moreover, fractionation of the water-extractable organic carbon showed that the ratio of hydrophobic to hydrophilic carbon increased to values greater than unity for stabilized compost. These results together with the analysis for non-cellulosic polysaccharides, phenolic compounds and organic nitrogen within the water extracts, confirmed the influence of solubilization, mineralization and organic matter transformation on the quality of the final compost.     

Microbial Community Growth and Utilization of Carbon Constituents During Thermophilic Composting at Different Oxygen Levels

Composting is characterized by dramatic changes in microbial community structure, to a high extent driven by changes in temperature and in the composition of the organic substrate. This study focuses on the interrelationships between decomposition of major classes in the organic material and dynamics in microbial populations during thermophilic composting of source-separated organic household waste. Experiments were performed in a 200-L laboratory reactor at 16, 2.5, and 1% O₂ in the compost atmosphere. Major classes of carbon constituents were analyzed by chemical methods, and the microbial biomass and community structure determined by fatty acid analyses with phospholipid fatty acids (PLFA) and total ester-linked fatty acids (EL) methods. At all three O₂ levels, the process was characterized by a rapid increase in microbial activity and biomass in the early thermophilic phase, although this period was delayed at the lower O₂ concentrations. Starch and fat were the main substrates utilized at all three O₂ levels during this period. The depletion of the starch fraction coincided with the beginning of a microbial biomass decrease, suggesting thatstarch is an important carbon substrate for the growth of thermophilic microorganisms during composting. Growth yields in the microbial community based on consumption of major carbon constituent classes in the high-activity period fell between 22 and 28%. Multivariate statistical analysis of changes in fatty acid composition revealed small, but statistically significant differences in the microbial community succession. At 16% O₂, 10Me fatty acids from Actinomycetes and cyclopropyl fatty acids (from Gram-negative bacteria) became more important with time, whereas 18:1ω7t was characteristic at 2.5 and 1% O₂, indicating a more stressed bacterial community at the lower O₂ concentrations. Although adequate composting was achieved at O₂ levels as low as 2.5 and 1%, it is not recommended to compost at such low levels in large-scale systems, because the heterogeneous gas transport through the material in these systems might lead to anaerobic conditions and inefficient composting.     

Differential response of high-elevation planktonic bacterial community structure and metabolism to experimental nutrient enrichment

Nutrient enrichment of high-elevation freshwater ecosystems by atmospheric deposition is increasing worldwide, and bacteria are a key conduit for the metabolism of organic matter in these oligotrophic environments. We conducted two distinct in situ microcosm experiments in a high-elevation lake (Emerald Lake, Sierra Nevada, California, USA) to evaluate responses in bacterioplankton growth, carbon utilization, and community structure to short-term enrichment by nitrate and phosphate. The first experiment, conducted just following ice-off, employed dark dilution culture to directly assess the impact of nutrients on bacterioplankton growth and consumption of terrigenous dissolved organic matter during snowmelt. The second experiment, conducted in transparent microcosms during autumn overturn, examined how bacterioplankton in unmanipulated microbial communities responded to nutrients concomitant with increasing phytoplankton-derived organic matter. In both experiments, phosphate enrichment (but not nitrate) caused significant increases in bacterioplankton growth, changed particulate organic stoichiometry, and induced shifts in bacterial community composition, including consistent declines in the relative abundance of Actinobacteria. The dark dilution culture showed a significant increase in dissolved organic carbon removal in response to phosphate enrichment. In transparent microcosms nutrient enrichment had no effect on concentrations of chlorophyll, carbon, or the fluorescence characteristics of dissolved organic matter, suggesting that bacterioplankton responses were independent of phytoplankton responses. These results demonstrate that bacterioplankton communities in unproductive high-elevation habitats can rapidly alter their taxonomic composition and metabolism in response to short-term phosphate enrichment. Our results reinforce the key role that phosphorus plays in oligotrophic lake ecosystems, clarify the nature of bacterioplankton nutrient limitation, and emphasize that evaluation of eutrophication in these habitats should incorporate heterotrophic microbial communities and processes.     

Dynamics of water-soluble carbon substances and microbial populations during the composting of de-inking paper sludge

Composting is an alternative method to dispose of de-inking paper sludge (DPS). Today, few studies have investigated the water-soluble carbon (WSC) substances as indicators of the decomposition process and the microbial changes taking place during the composting of DPS. Accordingly, the goal is to study their dynamics during the composting of DPS at three nitrogen levels, 0.6%, 0.7% or 0.9% total N, using mechanical turning. The changes in WSC substances, microbial biomass carbon (MBC) and, total and DPS microbial populations were monitored during 24 weeks. Also, microorganisms were identified and tested for the production of selected enzymes. Regardless of N treatments, the dynamic of WSC substances indicated that cellulose and hemicellulose fractions of DPS fibers were mainly biodegraded during the first 8 weeks while the more resistant carbon (C) fractions were biodegraded thereafter. MBC also evolved regardless of N treatments but was correlated to WSC substances. Its high values decline mostly after 12 weeks indicating the exhaustion of this source of C energy for microbial growth and the stabilisation of DPS organic matter. The dynamic and identified microorganisms were comparable to those observed in other composting processes. However, the results pointed out that those mostly implicated in the hydrolysis of DPS fibers were the thermophilic actinomycetes and fungi and, by comparison to the 0.6% or 0.7% N treatment, they decreased in presence of the 0.9% N treatment. Most microorganisms were hemicellulolytic bacteria, while actinomycetes and fungi were capable of degrading a wide variety of substrates. Overall, dynamics of WSC substances and microbial populations indicated that during composting, DPS decomposition obey a two phase decay while, contrary to the lowest N treatment, the 0.9% N treatment has slowed down this process by harming the important microbial populations implicated in the degradation of DPS fibers.     

蚯蚓堆制处理对农业有机废弃物的化学及生物学影响的主成分分析

在实验室可控条件下,以碳氮比28.7∶1的农业有机废弃物（牛粪和稻秆）为赤子爱胜蚓(Eisenia foetida)的培养基质,研究蚯蚓的堆制作用对有机物料的化学及生物学特性的影响.结果表明： 蚯蚓堆制处理30 d后,基质pH值、碳氮比显著降低,全磷显著升高,而全氮、碱解氮、可溶性碳、速效磷、微生物生物量碳、呼吸速率和微生物熵分别提高8.5%、2.6%、18%、63%、212%、44%和300%,有机质、呼吸熵分别降低5.0%和21.9%.蚯蚓堆制处理后物料具有较高的转化酶、酸性和碱性磷酸酶活性,较低的过氧化氢酶和脲酶活性.多元数据分析结果显示,自然堆制和蚯蚓堆制处理物料的化学和生物学特性均呈现显著的差异性.蚯蚓堆制处理优于自然堆制处理,可以明显改善有机物料的化学、生物学性质,是一种高效率处理农业有机废弃物的技术.     

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.     

Temporal variation in microbial and plant biomass during summer in a Mediterranean high-mountain dry grassland

Aims

We assessed the temporal changes on microbial biomass in relation to changes in soil moisture, dissolved organic carbon and plant biomass during the summer season in a Mediterranean high-mountain grassland.

Methods

Temporal variations were tested by two-way ANOVA. The relationships among microbial biomass, plant biomass, soil water content, soil organic carbon, dissolved organic carbon and total soil nitrogen during the summer season were assessed by means of structural equation modeling.

Results

Microbial biomass did not show variation, while dissolved organic carbon and root biomass decreased throughout the summer. Aboveground plant biomass peaked in the middle of the summer, when soil water content was at its minimum. Soil water content directly and negatively affected soil microbial biomass, and positively affected dissolved organic carbon. Moreover soil microbial biomass and dissolved organic carbon were negatively related. Plant biomass effects on soil microbial biomass were driven by root biomass, which indirectly affected soil microbial biomass through effects on soil organic carbon and soil nitrogen.

Conclusions

The temporal dynamic of microbial biomass during the summer season appeared to differ from previous observations in temperate alpine communities, and indicated the drought resistance of the microbial community during the summer in Mediterranean high-mountain grasslands. During the dry period, microbial biomass may play an alternative role in soil carbon conservation.     

An integrated mathematical model for co-composting of agricultural solid wastes with industrial wastewater

An integrated model for the composting process was developed. The structure of the model is such that it can be implemented in any mixture of different substrates, even in the case of co-composting of a solid waste with industrial wastewater. This paper presents a mathematical formulation of the physicochemical and biological principles that govern the composting process. The model of the co-composting ecosystem included mass transfer, heat transfer and biological processes. The biological processes included in the model were hydrolysis of particulate substrates, microbial growth and death. Two microbial populations (bacteria and fungi) were selected using Monod kinetics. Growth limiting functions of inhibitory factors, moisture and dissolved oxygen were added in the Monod kinetics. The bacteria were considered to utilise the easy biodegradable carbon hydrolysis product, fungi the difficult one, while both could degrade the carbon of wastewater. The mass balances of the most important nutrients, nitrogen and phosphorous, were also included in this approach. Model computer simulations provided results that fitted satisfactory the experimental data. Conclusively, the model could be a useful tool for the prediction of the co-composting process performance in the future and could be used to assist in the operation of co-composting plants.     

Impacts of drinking water pretreatments on the formation of nitrogenous disinfection by-products

The formation of disinfection by-products (DBPs), including both nitrogenous DBPs (N-DBPs) and carbonaceous DBPs (C-DBPs), was investigated by analyzing chlorinated water samples following the application of three pretreatment processes: (i) powdered activated carbon (PAC) adsorption; (ii) KMnO(4) oxidation and (iii) biological contact oxidation (BCO), coupled with conventional water treatment processes. PAC adsorption can remove effectively the precursors of chloroform (42.7%), dichloroacetonitrile (28.6%), dichloroacetamide (DCAcAm) (27.2%) and trichloronitromethane (35.7%), which were higher than that pretreated by KMnO(4) oxidation and/or BCO process. The removal efficiency of dissolved organic carbon by BCO process (76.5%)--was superior to that by PAC adsorption (69.9%) and KMnO(4) oxidation (61.4%). However, BCO increased the dissolved organic nitrogen (DON) concentration which caused more N-DBPs to be formed during subsequent chlorination. Soluble microbial products including numerous DON compounds were produced in the BCO process and were observed to play an essential role in the formation of DCAcAm in particular.     

Organic Layer Serves as a Hotspot of Microbial Activity and Abundance in Arctic Tundra Soils

Tundra ecosystem is of importance for its high accumulation of organic carbon and vulnerability to future climate change. Microorganisms play a key role in carbon dynamics of the tundra ecosystem by mineralizing organic carbon. We assessed both ecosystem process rates and community structure of Bacteria, Archaea, and Fungi in different soil layers (surface organic layer and subsurface mineral soil) in an Arctic soil ecosystem located at Spitsbergen, Svalbard during the summer of 2008 by using biochemical and molecular analyses, such as enzymatic assay, terminal restriction fragment length polymorphism (T-RFLP), quantitative polymerase chain reaction (qPCR), and pyrosequencing. Activity of hydrolytic enzymes showed difference according to soil type. For all three microbial communities, the average gene copy number did not significantly differ between soil types. However, archaeal diversities appeared to differ according to soil type, whereas bacterial and fungal diversity indices did not show any variation. Correlation analysis between biogeochemical and microbial parameters exhibited a discriminating pattern according to microbial or soil types. Analysis of the microbial community structure showed that bacterial and archaeal communities have different profiles with unique phylotypes in terms of soil types. Water content and hydrolytic enzymes were found to be related with the structure of bacterial and archaeal communities, whereas soil organic matter (SOM) and total organic carbon (TOC) were related with bacterial communities. The overall results of this study indicate that microbial enzyme activity were generally higher in the organic layer than in mineral soils and that bacterial and archaeal communities differed between the organic layer and mineral soils in the Arctic region. Compared to mineral soil, peat-covered organic layer may represent a hotspot for secondary productivity and nutrient cycling in this ecosystem.     

Dissolved Organic Matter Concentration and Quality Influences upon Structure and Function of Freshwater Microbial Communities

Past studies have suggested that the concentration and quality of dissolved organic matter (DOM) may influence microbial community structure. In this study, we cross-inoculated the bacterial communities from two streams and a dystrophic lake that varied in DOM concentration and chemistry, to yield nine fully crossed treatments. We measured dissolved organic carbon (DOC) concentration and heterotrophic microbial community productivity throughout a 72-h incubation period, characterized DOM quality by molecular weight, and determined microbial community structure at the initial and final time points. Our results indicate that all bacterial inoculate sources had similar effects upon DOC concentration and DOM quality, regardless of the DOM source. These effects included an overall decrease in DOM M _W and an initial period of DOC concentration variability between 0-24h. In contrast, microbial communities and their metabolic rates converged to profiles that reflected the DOM source upon which they were growing, regardless of the initial bacterial inoculation. The one exception was that the bacterial community from the low-concentration and low-molecular-weight DOM source exhibited a greater denaturing gradient gel electrophoresis (DGGE) band richness when grown in its own DOM source than when grown in the highest concentration and molecular weight DOM source. This treatment also exhibited a higher rate of productivity. In general, our data suggest that microbial communities are selected by the DOM sources to which they are exposed. A microbial community will utilize the low-molecular-weight (or labile) DOM sources as well as parts of the high-molecular-weight (refractory) DOM, until a community develops that can efficiently metabolize the more abundant high-molecular-weight source. This experiment examines some of the complex interactions between microbial community selection and the combined factors of DOM quality and concentration. Our data suggest that the roles of aerobic aquatic heterotrophic bacteria in carbon cycling, as well as the importance of high-molecular-weight DOM as a carbon source, may be more complex than is conventionally recognized.     

Influence of chronic ozone stress on carbon translocation pattern into rhizosphere microbial communities of beech trees (Fagus sylvatica L.) during a growing season

The influence of long-term chronic ozone exposure on carbon fluxes from young beech trees (Fagus sylvatica L.) into the phospholipid fraction of microbial communities (PLFA) in the rhizosphere and into the dissolved organic carbon (DOC) fraction was studied in a lysimeter experiment using ¹³C depleted CO₂ over one vegetation period to identify possible changes in below ground carbon translocation processes due to the plant stress. It could be shown that microbial biomass as well as individual microbial communities and their activity pattern in the rhizosphere of young beech trees are mainly driven by the vegetation period. An increase in total microbial biomass as well as individual microbial communities was detected during the vegetation period from June to September. However, also a clear ozone effect was visible mainly at the end of the vegetation period. Enzyme activities and PLFA data indicated earlier induced plant senescence as a response to the elevated ozone treatment. Furthermore higher microbial biomass and abundance of plant C utilizing microbes was observed in elevated ozone treatments over the whole vegetation period.