Evaluation of microbial population dynamics in the co-composting of cow manure and rice straw using high throughput sequencing analysis

Microbial population dynamics in co-composting of cow manure and rice straw were evaluated using 16S high throughput sequencing technology. Physicochemical factors, including temperature, pH, nitrogen contents, the ratio of carbon and nitrogen, and germination index, were also determined in this study. 16S high throughput sequencing results showed that bacterial community structure and composition significantly varied in each phase of composting. The major phyla included Bacteroidetes, Proteobacteria, Firmicutes, Actinobacteria and Planctomycetes, respectively. Bacteroidetes and Proteobacteria were the most abundant phyla in all phases, and Actinobacteria was just dominant in the mesophilic phase, while Firmicutes and Planctomycetes were ubiquitous. At the genus level, Simiduia, Flavobacterium, unclassified Chitinophagaceae and Flexibacter notably changed in each phase of composting. Bacterial community diversity in the mesophilic phase was higher than that in others based on the Shannon–Wiener index and Simpson diversity index. The ratio of carbon and nitrogen and germination index indicated that the co-composting of cow manure and rice straw reached maturation. The result of nitrogen contents showed that nitrogen loss mainly occurred in the thermophilic phase. In addition, the differences in the distributions of key OTUs between in the late thermophilic phase and the cooling and maturation phase were unobvious compared with other phase’s base on the principal component analysis. Redundancy analysis revealed that the changes of nitrogen played a predominant role in the distributions of OTUs during the composting process.     

Similarity of bacterial communities in sawdust- and straw-amended cow manure composts

We analyzed bacterial communities in two cow manure composts derived from the same feed manure and composted in the same location, but composted with different carbon amendments, and in peat-based potting mixes amended with these composts. Bacterial communities were characterized by PCR-denaturing gradient gel electrophoresis (DGGE) analysis of extracted DNAs, and population fingerprints generated for each sample were compared. Sequence analyses of dominant DGGE bands revealed that members of the phylum Bacteroidetes were the most dominant bacteria detected in this study (19 of 31 clones). These analyses demonstrate that bacterial community profiles of individual composts were highly similar, as were profiles of compost-amended potting mixes. However, potting mix profiles differed substantially from the original compost profiles and from that of the peat base. These data indicate that highly similar bacterial populations were active in the two composts, and suggest that the effects of the initial carbon amendment on the mature compost bacterial communities were minor, while factors such as the feed manure and composting location may have been more influential.     

Impact of fertilization by natural manure on the microbial quality of soil: Molecular approach

The quality of soil is strongly bound by several interactions between chemical and biological components, including microbial composition, which are a key importance for soil performance. Cultural activities have a huge induction on soil health, through both modification of physicochemical proprieties and changing on soil microbial communities. This usually affects the safety of soil, and then the crop production and water.In the present work, the information on bacterial community composition was determined from a set of 6 soils collected from 2 farms in agricultural land of Marrakech (Morocco), one of which used poultry manure (PM) and the other cow manure (CM) as fertilizers. To profile this structure of the bacterial community Denaturing Gradient Gel Electrophoresis (DGGE) of 16S rDNA fragments has been used.These amendments resulted in the appearance of several novel bands and different relative intensities of bands between the control station and other sites studied. The stations most affected are those receiving a supply of manure rather high, which results in an organic and bacterial load in the soil. The results showed a bacterial diversity very important indicating a fecal contamination like Bacteroides, Pseudomonas, Staphylococcus,… etc. Bacteria pertain to the phylum Firmicutes and Bacteroidetes were noted to be the dominant ribotype in amended soil.Moreover, this work demonstrates also the existence of pathogens strains in soil amended by poultry manure (PM) belonging to the Clostridiales order and Pseudomonadales. The pathogenic bacteria detected posing a hazard of human contagion when they are used for soil practice.     

Changes in Bacterial and Fungal Communities across Compost Recipes,Preparation Methods,and Composting Times

Compost production is a critical component of organic waste handling, and compost applications to soil are increasingly important to crop production. However, we know surprisingly little about the microbial communities involved in the composting process and the factors shaping compost microbial dynamics. Here, we used high-throughput sequencing approaches to assess the diversity and composition of both bacterial and fungal communities in compost produced at a commercial-scale. Bacterial and fungal communities responded to both compost recipe and composting method. Specifically, bacterial communities in manure and hay recipes contained greater relative abundances of Firmicutes than hardwood recipes with hay recipes containing relatively more Actinobacteria and Gemmatimonadetes. In contrast, hardwood recipes contained a large relative abundance of Acidobacteria and Chloroflexi. Fungal communities of compost from a mixture of dairy manure and silage-based bedding were distinguished by a greater relative abundance of Pezizomycetes and Microascales. Hay recipes uniquely contained abundant Epicoccum, Thermomyces, Eurotium, Arthrobotrys, and Myriococcum. Hardwood recipes contained relatively abundant Sordariomycetes. Holding recipe constant, there were significantly different bacterial and fungal communities when the composting process was managed by windrow, aerated static pile, or vermicompost. Temporal dynamics of the composting process followed known patterns of degradative succession in herbivore manure. The initial community was dominated by Phycomycetes, followed by Ascomycota and finally Basidiomycota. Zygomycota were associated more with manure-silage and hay than hardwood composts. Most commercial composters focus on the thermophilic phase as an economic means to insure sanitation of compost from pathogens. However, the community succeeding the thermophilic phase begs further investigation to determine how the microbial dynamics observed here can be best managed to generate compost with the desired properties.     

New insights into the structure and dynamics of actinomycetal community during manure composting

Despite advancing knowledge about the functional role of actinomycetes in degrading lignocellulosic materials, definitive knowledge concerning the diversity and dynamics of the actinomycetal community in composting is still lacking. In this study, real-time polymerase chain reaction (PCR) coupled with denaturing gradient gel electrophoresis (DGGE) and clone library construction were applied to investigate actinomycetal diversity and dynamics in a pilot-scale composting. Quantitative real-time PCR data revealed that actinomycetes accounted for 18–86 % of bacteria and that the fraction peaked during the maturing phase, indicating that Actinobacteria were critical to the compost ecosystem. Qualitatively, actinomycetal communities displayed distinct temporal variations during composting. Fourteen distinct genera of actinomycetes and an unknown group were observed in manure composts. Redundancy analysis indicated that temperature exerted an influence over the actinomycetal communities. Specifically, pathogenic Corynebacterium species dominated in the initial phase, whereas the genera Saccharomonospora and Thermobifida were abundant in the thermophilic phase. In maturing composts, mesophilic Micrococcineae members were most prevalent. The dominant thermophiles along with Micrococcineae may jointly facilitate the degradation of lignocellulosic materials during composting. Together, our research revealed a more detailed ecological and potential functional role for actinomycetes in the compost ecology.     

Co-composting solid swine manure with pine sawdust as organic substrate

The main objectives of this work were to investigate the evolution of the principal physicochemical properties, i.e., bulk temperature, pH, electrical conductivity (EC), moisture content, total organic matter, total nitrogen and total phosphorus, in co-composting pine sawdust with increasing percentages of fresh solid swine manure, and thus to evaluate the most desirable manure proportion for producing organic substrates in consideration of the quality of the resulted compost. The composting was in four identical 100.5l lab vessels, using 5% each tea leaves and herb residues as conditioners. The swine manure was added in the trials at 20%, 30%, 40%, respectively, and was substituted in the control with 30% lake sludge corrected by 0.5% urea. The initial humidity of each treatment was 60+/-2%. While being aerated actively at approximately 0.3m(3)/min at intervals of 10 min/h, the mixture was composted for 29 days. The results indicated that N and P decomposition primarily occurred in the mesophilic phase, while organic carbon decomposed in the thermophilic phase and 30% swine manure with initial C/N ratio of about 40 was more desirable for composting organic substrates.     

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。     

Effects of fertilization on bacterial community structure and function in a black soil of Dehui region estimated by Biolog and PCR-DGGE methods

Soil microbial community structure and function are commonly used as indicators for soil quality and fertility. In this paper, the bacterial community structure and function in a black soil of Dehui region influenced by fertilization were investigated by Biolog and PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) methods. Biolog examination showed that substrate richness and catabolic diversities of bacterial communities were the highest in the treatment of farm yard manure, and the lowest in the chemical fertilizer treatment. DGGE fingerprint showed that the majority of bands were similar among all treatments, suggesting that microbial communities with those bands were stable, and not influenced by fertilization. In general, chemical fertilizer decreased the diversity of soil bacterial communities. The PCA (principal component analysis) plots of Biolog and DGGE revealed that the structure and function of bacterial communities were similar in the non-fertilized control and the treatment of farm yard manure alone, which inferred that the application of farm yard manure increased the quantity of soil microbes but had less effect on the changes of community structure. The catabolic function was similar, but the composition structure differed between the treatments of chemical fertilizer alone and combined application of farm yard manure with chemical fertilizer. These results suggest that the use of chemical fertilizer mainly decreased the catabolic activity of the fast growth bacteria or eutrophic bacteria.     

Effect of fungal consortium and animal manure amendments on phosphorus fractions of paddy-straw compost

The study was conducted to reveal the type of phosphorus (P) fractions present in mature compost prepared by co-composting paddy straw (P.S) with cattle manure (CM), farm yard manure (FYM) and poultry manure (PM), each added separately as nitrogen (N) and P source. A consortium of phytate mineralizing fungi developed by including Aspergillus niger ITCC 6719, Aspergillus flavus ITCC 6720 and Trichoderma harzianum ITCC 6721 was applied for recovery of P from plant and animal residues. Chemical evaluation of compost after 4 months of aerobic decomposition revealed that inoculation improved the sodium bicarbonate-extractable P content of CM and FYM supplemented P.S compost by 32.3% and 23.5% respectively compared with their respective un-inoculated control. However, the peak values for water soluble-P fractions were recorded in CM–straw compost followed by PM–straw compost. Fungal inoculation also improved the agronomic quality of PM–straw compost as the latter had the highest total P content and lowest C:N and E4/E6 ratio of 18:1 and 5.36:1 respectively. The recovery of organic P from agricultural residue has the potential to reduce the application of synthetic P fertilizer. P-enriched organic manure can offer potential environment and economic benefits to farmers under sustainable agriculture.     

Changes in bacterial communities during two agricultural solid wastes’ co-composting processes

Microbes perform an important role in the solid-state fermentation (SSF) process, and bacterial communities are more or less abundant depending on the starting materials and the composting procedure. In this study, high-throughput sequencing was used to investigate the changes in bacterial communities in different composting piles containing spent pig litter and distiller grains in five dry weight ratios, i.e., 100% distiller grains (treatment 1), 75% distillers grains/25% spent pig litter (treatment 2), 50% distillers grains/50% spent pig litter (treatment 3), 25% distillers grains/75% spent pig litter (treatment 4), and 100% spent pig litter (treatment 5). The results showed that the fermentation time of the thermophilic stage was prolonged with an appropriate content of distiller grains. The alpha-diversity analysis showed that the variation in bacterial richness and diversity in the various treatments (except treatment 5) was greater in the high-temperature stage than in the inception stage and then was slightly lower in the stabilization stage than in the thermophilic stage. The relative abundance of the predominant bacterial communities differed during different composting stages except in treatment 5. The abundant bacterial communities were similar among the treatments with different proportions of distiller grains (treatments 2, 3, and 4) during the high-temperature stage but differed during the stabilization stage, as increasing proportions of distiller grains increased the relative abundance of the phyla Proteobacteria and Firmicutes, and decreased that of Actinobacteria. Additionally, principal coordinate analysis (PCoA) showed that the bacterial communities in treatments 1, 2, 3, and 4 during the initial stage (day 0) were different from those observed during two other stages (day 10 and day 53), while treatment 5 showed only slight variations in the bacterial community structure in response to changes in the composting process. The results indicated that spent pig litter is not suitable for single-material composting and the addition of an appropriate amount of distiller grains can improve the fermentation process. The understanding of the microbial community diversity at molecular level provided a theoretical basis for the optimization of spent pig litter and/or distiller grain fermentation.     

Characterization of dairy cattle manure/wallboard paper compost mixture

The aim of this research was to evaluate the use of manufacturing wallboard paper scraps as an alternative bulking agent for dairy cattle manure composting. The characteristics of the composting process were studied based on the changes in physico-chemical parameters and final compost quality. Composting of dairy cattle manure with wallboard paper was performed in a 481-L cylindrical reactor with vacuum-type aeration. Rapid degradation of organic matter was observed during the thermophilic stage of composting due to high microbial activity. High temperature and alkaline pH conditions promoted intense ammonia emission during the early stage of composting. The number of mesophilic and thermophilic microorganisms were found to be affected by changes in temperature at different composting stages. The total nitrogen (N), phosphorus (P), potassium (K), and sodium (Na) concentrations of the mixture did not change significantly after 28days of composting. However, the presence of gypsum in the paper scraps increased the calcium content of the final compost. The wallboard paper had no phyto-inhibitory effects as shown by high germination index of final compost (GI=99%).     

Effect of low initial C/N ratio on aerobic composting of swine manure with rice straw

Two pilot composting experiments were conducted to investigate the effect of low initial C/N ratio on the composting of swine manure with rice straw by measuring physical and chemical parameters. The results showed that the thermophilic duration of bin 1 and bin 2 was long enough to satisfy the sanitary standard, and swine manure could reach maturity. Bin 1 containing larger amount of swine manure and less amount of rice straw showed a higher nitrogen loss (8%), shorter thermophilic phase, and longer maturity time (about 2 weeks) than bin 2. However, economical analysis showed a lower initial C/N ratio (20) could reduce 172 kg rice straw per ton fresh swine manure than a higher C/N ratio (25), and more swine manure could also be treated. Therefore, a low initial C/N ratio (20) could be suggested in the composting of swine manure with rice straw.     

Effects of Continuous Thermophilic Composting (CTC) on Bacterial Community in the Active Composting Process

The method of continuous thermophilic composting (CTC) remarkably shortened the active composting cycle and enhanced the compost stability. Effects of CTC on the quantities of bacteria, with a comparison to the traditional composting (TC) method, were explored by plate count with incubation at 30, 40 and 50°C, respectively, and by quantitative PCR targeting the universal bacterial 16S rRNA genes and the Bacillus 16S rRNA genes. The comparison of cultivatable or uncultivatable bacterial numbers indicated that CTC might have increased the biomass of bacteria, especially Bacillus spp., during the composting. Denaturing gradient gel electrophoresis (DGGE) analysis was employed to investigate the effects of CTC on bacterial diversity, and a community dominated by fewer species was detected in a typical CTC run. The analysis of sequence and phylogeny based on DGGE indicated that the continuously high temperature had changed the structure of bacterial community and strengthened the mainstay role of the thermophilic and spore-forming Bacillus spp. in CTC run.     

Organic matter evolution during co-composting of the organic fraction of municipal waste and poultry manure

The study concerned the evolution of organic matter, and the humification process, during the co-composting of the organic fraction of municipal solid waste (OMSW) and poultry manure (PM); the study was made with two different mixtures (OMSW:PM ratios of 3:2 and 2:3, wet weight:wet weight) and two different particle sizes (1 and 0.2cm). The results suggested that the composting process proceeded unhindered throughout the degradation of easily degradable materials like hemicellulose, and that of the rather less degradable cellulose and lipids, and the concentration of recalcitrant material, i.e. a ligno-humic (LU) fraction. These processes were more evident for mixtures with lower particle size. Throughout the composting, in all mixtures studied, humification proceeded by the formation of a new HA fraction, which was probably the result of the partial degradation and solubilization of more complex insoluble organic molecules, i.e. humin fraction.     

Assessing the impact of composting and vermicomposting on bacterial community size and structure, and microbial functional diversity of an olive-mill waste

The aim of this study was to couple biochemical and molecular methodologies for evaluating the impact of two recycling technologies (composting and vermicomposting) on a toxic organic waste. To do this, six enzyme activities controlling the key metabolic pathways of the breakdown of organic matter, real-time PCR assays targeting 16S rRNA genes, and denaturing gradient gel electrophoresis (DGGE) profiling-sequence analysis of PCR-amplified 16S rRNA fragments have been used to determine the functional diversity, bacterial number, and bacterial community structure, respectively, in a mixture of olive waste and sheep manure, and in the derived compost and vermicompost. Both the recycling technologies were effective in activating the microbial parameters of the toxic waste, the vermicomposting being the best process to produce greater bacterial diversity, greater bacterial numbers and greater functional diversity. Although several identical populations were detected in the processed and non-processed materials, each technology modified the original microbial communities of the waste in a diverse way, indicating the different roles of each one in the bacterial selection.     

Species Diversity and Substrate Utilization Patterns of Thermophilic Bacterial Communities in Hot Aerobic Poultry and Cattle Manure Composts

This study investigated the species diversity and substrate utilization patterns of culturable thermophilic bacterial communities in hot aerobic poultry and cattle manure composts by coupling 16S rDNA analysis with Biolog data. Based on the phylogenetic relationships of 16S rDNA sequences, 34 thermophilic (grown at 60 degrees C) bacteria isolated during aerobic composting of poultry manure and cattle manure were classified as Bacillus licheniformis, B. atrophaeus, Geobacillus stearothermophilus, G. thermodenitrificans, Brevibacillus thermoruber, Ureibacillus terrenus, U. thermosphaericus, and Paenibacillus cookii. In this study, B. atrophaeus, Br. thermoruber, and P. cookii were recorded for the first time in hot compost. Physiological profiles of these bacteria, obtained from the Biolog Gram-positive (GP) microplate system, were subjected to principal component analysis (PCA). All isolates were categorized into eight different PCA groups based on their substrate utilization patterns. The bacterial community from poultry manure compost comprised more divergent species (21 isolates, seven species) and utilized more diverse substrates (eight PCA groups) than that from cattle manure compost (13 isolates, five species, and four PCA groups). Many thermophilic bacteria isolated in this study could use a variety of carboxylic acids. Isolate B110 (from poultry manure compost), which is 97.6% similar to U. terrenus in its 16S rDNA sequence, possesses particularly high activity in utilizing a broad spectrum of substrates. This isolate may have potential applications in industry.     

Effects of Manure Compost Application on Soil Microbial Community Diversity and Soil Microenvironments in a Temperate Cropland in China

The long-term application of excessive chemical fertilizers has resulted in the degeneration of soil quality parameters such as soil microbial biomass, communities, and nutrient content, which in turn affects crop health, productivity, and soil sustainable productivity. The objective of this study was to develop a rapid and efficient solution for rehabilitating degraded cropland soils by precisely quantifying soil quality parameters through the application of manure compost and bacteria fertilizers or its combination during maize growth. We investigated dynamic impacts on soil microbial count, biomass, basal respiration, community structure diversity, and enzyme activity using six different treatments [no fertilizer (CK), N fertilizer (N), N fertilizer + bacterial fertilizer (NB), manure compost (M), manure compost + bacterial fertilizer (MB), and bacterial fertilizer (B)] in the plowed layer (0–20 cm) of potted soil during various maize growth stages in a temperate cropland of eastern China. Denaturing gradient electrophoresis (DGGE) fingerprinting analysis showed that the structure and composition of bacterial and fungi communities in the six fertilizer treatments varied at different levels. The Shannon index of bacterial and fungi communities displayed the highest value in the MB treatments and the lowest in the N treatment at the maize mature stage. Changes in soil microorganism community structure and diversity after different fertilizer treatments resulted in different microbial properties. Adding manure compost significantly increased the amount of cultivable microorganisms and microbial biomass, thus enhancing soil respiration and enzyme activities (p<0.01), whereas N treatment showed the opposite results (p<0.01). However, B and NB treatments minimally increased the amount of cultivable microorganisms and microbial biomass, with no obvious influence on community structure and soil enzymes. Our findings indicate that the application of manure compost plus bacterial fertilizers can immediately improve the microbial community structure and diversity of degraded cropland soils.     

施肥和土壤管理对土壤微生物生物量碳、氮和群落结构的影响

以中国科学院沈阳生态试验站的长期定位试验为平台,研究了不同施肥和土壤管理对潮棕壤微生物生物量碳、氮和群落结构的影响。结果表明,裸地和农田处理的微生物生物量碳、氮较低,但是农田处理下施肥增加了微生物生物量,其中NPK+M效果最明显。DGGE图谱显示,处理间细菌条带分布较相似,其中裸地的细菌多样性最高;长期施肥和土壤管理改变了土壤真菌群落结构,施肥增加了真菌多样性,且有机肥的影响大于化肥;不同处理间氨氧化细菌群落结构差异显著,NPK+M显著增加了氨氧化细菌多样性,且无机肥和有机肥对氨氧化细菌群落影响不同。施肥和土壤管理对细菌影响较小,但显著改变了真菌和氨氧化细菌的群落结构。聚类分析结果显示,土壤管理措施较施肥对细菌、真菌和氨氧化细菌群落的影响更为显著。