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.     

Microbiological parameters as indicators of compost maturity

AIMS: The objectives of this study were to determine the changes of microbial properties of pig manure collected from pens with different management strategies and composted using different turning and moisture regimes; relate their association with humification parameters and compost temperature; and identify the most suitable microbial indicators of compost maturity. METHODS AND RESULTS: Six different microbial parameters, including total bacterial count, oxygen consumption rate, ATP content, dehydrogenase activity, and microbial biomass C and N, along with humification parameters [humic acid (HA), fulvic acid (FA) and HA : FA ratio] and compost temperature were monitored during composting. Significant positive correlations were found between temperature and microbial properties, including O2 consumption rate, ATP content, dehydrogenase activity, and microbial biomass N. The humification parameters also showed significant correlations with microbial properties of the manure compost. For instance, HA contents of pig manures was positively correlated with total aerobic heterotrophs, and microbial biomass N and C; and negatively correlated with O2 consumption rate, ATP content, and dehydrogenase activity. Among the six microbial parameters examined, dehydrogenase activity was the most important factor affecting compost temperature and humification parameters. Composting strategies employed in this study affected the speed of composting and time of maturation. If the moisture content is maintained weekly at 60% with a 4-day turning frequency, the pig manure will reach maturity in 56 days. CONCLUSIONS: The composting process went through predictable changes in temperature, microbial properties and chemical components despite differences in the initial pig manure and composting strategies used. Among the six microbial parameters used, dehydrogenase activity is the most suitable indicator of compost maturity. Compared with respiration rate, ATP content and microbial biomass procedures, dehydrogenase activity is the simplest, quickest, and cheapest method that can be used to monitor the stability and maturity of composts. SIGNIFICANCE AND IMPACT OF THE STUDY: The results presented here show that microbial parameters can be used in revealing differences between composts and compost maturity. The statistical relationship established between humification parameters and microbial parameters, particularly dehydrogenase activity, demonstrates that it is possible to monitor the composting process more easily and rapidly by avoiding longer and more expensive analytical procedures.     

畜禽粪便堆肥过程中各种氮化合物的动态变化及腐熟度评价指标

对不同畜禽粪便在堆肥过程中各种含氮化合物的动态变化进行了研究,结合综合性腐熟度评价指标——种子发芽指数（GI）,探讨了畜禽粪便堆肥过程中与氮有关的腐熟度评价指标.结果表明：随着堆肥的进行,除奶牛粪外,其它畜禽粪便的全氮（TN）含量均呈先下降而后平稳变化趋势,奶牛粪则呈先增加而后平稳变化趋势;各种畜禽粪便中,碱解性氮（HN）含量先增后降;NH₄⁺-N含量先下降而后保持平稳;NO₃^- -N含量则持续增加;NH₄⁺ -N/NO₃^- -N迅速降低.堆肥腐熟度指标中,除综合性评价指标GI值外,HN/TN和NH₄⁺ -N/TN也可作为评价畜禽粪便腐熟程度的优选指标,而NO₃^- -N/TN只能作为一般性评价指标.根据综合性评价指标GI值达到腐熟要求的标准（GI>0.50）,除仔猪粪外,其它畜禽粪便在HN/TN<20.77％、NH₄⁺ -N/TN<10.06％及NO₃^- -N/TN>0.38％时基本达到腐熟要求.     

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%).     

有机固体废弃物堆肥的物质变化及腐熟度评价

堆肥是资源化处理有机固体废弃物的有效途径之一．有机固体废弃物的种类繁多,物科的性质差异很大,另外,由于堆肥条件的不同,也会引起堆肥的物质变化有很大差异．堆肥的稳定度和腐熟度是衡量堆肥产品质量的尺度,其评价对于安全农用有着重要意义．稳定度则重于堆肥施用对周围环境的影响,而腐熟度则重于堆肥施用对植物生长的影响,它们受堆肥物料,堆肥条件等诸多因素的综合影响,其评价指标因而多种多样．堆肥稳定度主要从堆肥的温度、颜色、CO2是最为简便的稳定度评价指标,当其趋于环境温度时,表明堆肥达到稳定．堆肥腐熟度的评价指标很多,包括化学指标、生物活性指标以及植物毒性指标3类,其中种子发芽系数作为植物毒性指标被认为是最可靠的评价指标之一。     

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

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

Effect of inoculating white-rot fungus during different phases on the compost maturity of agricultural wastes

The lignocellulolytic microorganism, Phanerochaete chrysosporium, was inoculated during different phases of agricultural wastes composting, and its effect on compost maturity was studied. In the three runs, the decrease in C/N ratio and increases in germination index and humification indices (humification ratio, humification index, percentage of humic acids and degree of polymerisation) were found. Furthermore, the different effects of inoculation during different phases on the compost maturity could be observed by using ANOVA. When inoculated during the second fermentation phase, P. chrysosporium induced significant changes on all parameters of compost maturity except C/N ratio, whereas it did not produce an obvious change on these parameters when inoculated during the first fermentation phase.     

Comparison of signature lipid methods to determine microbial community structure in compost

The microbial community structure changes substantially during the composting process and simple methods to follow these changes can potentially be used to estimate compost maturity. In this study, two such methods, the microbial identification (MIDI) method and the ester-linked (EL) procedure to determine the composition of long-chain fatty acids, were applied to compost samples of different age. The ability of the two methods to describe the microbial succession was evaluated by comparison with phospholipid fatty acid (PLFA) analysis on the same samples.Samples were taken from a 200-l laboratory compost reactor, treating source-separated organic household waste. During the initial stages of the process, the total concentration of fatty acids in compost samples treated with the EL and MIDI methods was many times higher than with the PLFA method. This was probably due to the presence of fatty acids from the organic material in the original waste. However, this substantial difference between PLFA and the other two methods was not found later in composting. Although the PLFA method gave the most detailed information about the growth and overall succession of the microbial community, the much simpler MIDI and EL methods also successfully described the shift from the initially dominating straight chain fatty acids to iso- and anteiso branched, 10 Me branched and cyclopropane fatty acids in the later stages of the process. Thus, the MIDI and EL extraction methods appear to be suitable for analysis of microbial FAME profiles in compost, particularly in the later stages of the process.     

Heavy metal contamination of a mixed waste compost: Metal speciation and fate

The aims of this study were to assess changes in heavy metal availability in two contrasting feedstocks during aerobic composting, and the availability of said metals in the finished composts. A high C-to-N ratio mixed biodegradable municipal solid waste (MSW) feedstock was successfully composted on its own and in combination with green waste. Changes in heavy metal speciation throughout the composting process were studied using the modified BCR sequential extraction protocol. It was found that total Cu, Pb and Zn concentrations increased over time due to the progressive mineralization of the compost feedstock. Metals were fractionated differently within the two feedstocks, although only Cu showed significant redistribution (mostly to the oxidisable fraction) over the 5 month composting period. The MSW-derived composts performed comparably with other commercially-available composts in a series of plant growth trials. Plant metal accumulation was not influenced by the heavy metals present in the MSW-derived compost implying that they are not plant available. It is recommended that these relatively low value/quality composts may be used for remediation of acidic heavy metal contaminated sites.     

A modified static respiration assay and its relationship with an enzymatic test to assess compost stability and maturity

Despite the numerous compost stability and maturity tests, no universally accepted compost stability or maturity index exists. The fluorescein di-acetate (FDA) enzymatic assay, originating from soil studies, is examined here as a potential new compost stability test, and is compared to microbial respiration and phytotoxicity indices. Thirteen composts were used in the study from different source materials. Static microbial respiration activity indices calculated were the cumulative O₂ consumptions, O₂ consumption rates, total C-CO₂ production, the respiratory quotient and the bio C/N ratio. Compost phytotoxicity was quantified via a 7-day tomato seed germination assay. Results showed that the net fluorescein release rates correlated with all stability indices. The germination index marginally correlated with the fluorescein release rates, but not with any of the other stability indices. New limits to classify composts regarding their stability were proposed.     

Comparison of characterization and microbial communities in rice straw- and wheat straw-based compost for <Emphasis Type="Italic">Agaricus bisporus</Emphasis> production

Rice straw (RS) is an important raw material for the preparation of Agaricus bisporus compost in China. In this study, the characterization of composting process from RS and wheat straw (WS) was compared for mushroom production. The results showed that the temperature in RS compost increased rapidly compared with WS compost, and the carbon (C)/nitrogen (N) ratio decreased quickly. The microbial changes during the Phase I and Phase II composting process were monitored using denaturing gradient gel electrophoresis (DGGE) and phospholipid fatty acid (PLFA) analysis. Bacteria were the dominant species during the process of composting and the bacterial community structure dramatically changed during heap composting according to the DGGE results. The bacterial community diversity of RS compost was abundant compared with WS compost at stages 4–5, but no distinct difference was observed after the controlled tunnel Phase II process. The total amount of PLFAs of RS compost, as an indicator of microbial biomass, was higher than that of WS. Clustering by DGGE and principal component analysis of the PLFA compositions revealed that there were differences in both the microbial population and community structure between RS- and WS-based composts. Our data indicated that composting of RS resulted in improved degradation and assimilation of breakdown products by A. bisporus, and suggested that the RS compost was effective for sustaining A. bisporus mushroom growth as well as conventional WS compost.     

Characterization of compost produced from separated pig manure and a variety of bulking agents at low initial C/N ratios

The aim of this study was to investigate the composting of separated pig manure solids with or without a variety of bulking agents at a low initial C/N ratio (12.5-23.3). Compost stability was investigated using an oxygen uptake rate (OUR) test and compost maturity was investigated using a germination index test. All treatments showed typical patterns of compost temperature. Temperatures above 60 °C were achieved by Day 2, followed by a thermophilic phase (50-60 °C), which lasted for 1 to 2 weeks followed by a cooling phase. The stability of one of treatments which did not contain any bulking agent - OUR of 25 mmol O₂ kg⁻¹ OM hour⁻¹ - was negatively affected by its initial high water content (69%). The addition of a bulking agent and initial water content below 60% were necessary to compost the separated solid fraction of pig manure at a low initial C/N ratio.     

Effect of chitin waste-based composts produced by two-phase composting on two oomycete plant pathogens

Biphasic composts were prepared by first mixing peat moss and sawdust with a nitrogen-rich biomass such as chitinous waste or cow manure and composting them until termination of the thermophilic phase. These partially stabilized composts were then amended with shrimp waste inducing a second thermophilic phase. Filter-sterilized water extracts obtained from two mature biphasic composts (SP₂W₂+S and MPW+S) reduced the growth of two oomycete plant pathogens, Phytophthora fragariae var. rubi and Pythium ultimum. Both SP₂W₂+S and MPW+S composts significantly reduce the incidence of cucumber damping-off caused by Pythium ultimum as compared to a commercial brand of compost made from shrimp waste and peat moss. Hydrolysis products of chitin were unlikely to be responsible for growth inhibition since no oligomeric forms of chitin were detected in SP₂W₂+S. The shrimp waste amendment carried out after the first thermophilic phase modified the microbial populations of biphasic composts. Following the amendment, the proportion of branched-chain microbial fatty acids typical of Gram-positive bacteria increased considerably suggesting that this group of bacteria became more prevalent within the total microbial population. These data suggests that the two-phase composting process promotes the proliferation of Gram-positive bacteria antagonistic to oomycete plant pathogens.     

Aerobic composting of chips from clear-cut trees with various co-materials

Swollen chips made from trees felled during clear-cutting were composted with various organic and inorganic materials in an aerobic composting reactor for 5 months and then piled for 5 months. The organic materials included chicken feces, urea, nitrogenous lime (calcium cyanamide, manure), and material rapidly composted from food garbage in 24-h bacterial fermentation, while the inorganic materials were coal ash and volcanic ash. In this paper, we first attempt to estimate the quality and degree of maturity of each compost from its chemical properties. Furthermore, we try to calculate the maturity of the fermented wood chip composts from their mixture ratio of the initial materials by multiple linear regression analysis. We measured changes in the C/N ratio, nitrate nitrogen (NO3-N) content, percentage of humic acid in the alkali soluble fraction (PQ), cation exchange capacity, pH, and EC during the composting period. The degrees of maturity of the composts were estimated via a plant growth test using Chinese cabbage. We found that the CN ratio, NO3-N concentration, and PQ were suitable for estimating the degree of maturity of wood chip composts. For maturity, the CN ratio should be less than 14, the PQ more than 66.2, and the NO3-N concentration more than 853 mgkg-1. We devised an equation to estimate the degree of maturity after 10 months by a multiple linear regression analysis from the mixing ratio of wood chips and the co-composted materials. From the multiple linear regression analysis, the above three indices of compost maturity could be estimated from the mixing ratio of the initial materials. This equation should enable us to determine the degree of compost maturity after 10 months based on the initial mixing ratio.     

湿度对堆肥理化性质的影响

水分是堆肥微生物生命活动的基础 ,也是堆肥中重要的工艺控制参数。弄清湿度对堆肥微生物及理化性质的影响 ,对于优化堆肥工艺参数、提高堆肥效率、降低投资和运行成本具有重要意义。综述了堆肥湿度研究的动态 ,指出了当前研究中存在的问题 ,并提出了未来的研究方向。大量的研究表明 ,湿度低于 4 5 %或高于 6 5 %都不利于堆肥处理。湿度太高会导致堆料的压实度增加、FAS减少、透气性能降低 ,从而导致堆体内氧气供应不足、堆肥升温困难、有机物降解速率降低、堆肥周期延长。湿度过低 ,水分会限制堆肥微生物的新陈代谢 ,导致微生物活性下降、堆肥腐熟困难。由于鼓风、散热、水蒸发等会使堆体内存在湿度的空间变异 ,也会降低堆肥效率和堆肥产品的质量。另外 ,堆肥湿度还影响堆肥的保肥能力。由各文献得出结论 ,堆肥的最佳湿度范围一般为 5 0 %～ 6 0 %左右     

Microbial studies of compost: bacterial identification,and their potential for turfgrass pathogen suppression

Composting is the degradation of organic materials through the activities of diverse microorganisms. This research examined microbial community dynamics, population levels and identification of bacteria throughout the composting process and in storage. In addition, an evaluation was performed to determine the potential for dominant bacterial isolates to suppress selected turfgrass pathogens: Sclerotinia homoeocarpa, Pythium graminicola, Typhula ishikariensis, and Microdochium nivale, responsible for causing the turfgrass diseases dollar spot, pythium blight, typhula blight, and fusarium patch, respectively. Composts supported high population levels of bacteria with 78% of cultures tested being Gram-negative. Proteolytic activity, found in 29% of cultures tested is a potential mechanism of suppression or competition with other microorganisms. Although the Biolog system did not identify a wide range of bacteria, the main Gram-negative genera identified in mature compost were Pseudomonas (28%), Serratia (20%), Klebsiella (11%), and Enterobacter (5%). Twenty-one percent of isolates tested were not identified by Biolog, and many more had similarity indexes < 0.50. The microbial identification system, based on whole cell fatty acid analysis, identified a wide range of bacteria, with a higher proportion of similarities than the Biolog system. Genera common to both testing procedures included Pseudomonas, Serratia, and Enterobacter. All Gram-positives were identified as Bacillus spp. Phospholipid fatty acid analysis, used to estimate the diversity of microbial communities, was useful in monitoring changes in microbial population in storage and during composting, as well as estimating levels of compost maturity. Plate challenge experiments revealed a number of cultures with antagonistic activity against turfgrass pathogens. There were 52, 31, 32 and 19% of the bacterial isolates tested that exhibited antagonistic activity against S. homoeocarpa, P. graminicola, T. ishikariensis, and M. nivale, respectively. Improved understanding of microbial populations and their dynamics in composts will expand their potential to act as suppressants on pathogenic fungi or turfgrass.     

Composting of bio-waste, aerobic and anaerobic sludges--effect of feedstock on the process and quality of compost

In-vessel composting of three stocks with originally different degree of organic matter degradation was conducted for: (1) kitchen source-separated bio-waste (BW), (2) aerobic (AS) as well as (3) anaerobic sludges (AnS) from municipal wastewater treatment plant. Composting experiment lasted over a year. The highest activity of the process was in the BW compost. It was implied by the highest temperature, CO(2) release, ammonification and nitrification, intensive accumulation and removal of low-weight carboxylic acids (water- and NaOH-extractable). Between the sludges higher mineralization and CO2 release was in AnS, while ammonification and nitrification were higher in AS compost; no significant difference between sludge composts was noticed for dynamics of pH, conductivity, concentrations of LWCA, and some nutrient compounds and heavy metals. Nitrogen content of the final compost increased in BW, but decreased in AS and AnS. Phytotoxicity of Lepidium sativum was eliminated faster in sludge composts compared to BW compost.     

Assessment of survival of Listeria monocytogenes,Salmonella Infantis and Enterococcus faecalis artificially inoculated into experimental waste or compost

Aims: To evaluate survival of pathogenic strains, Listeria monocytogenes and Salmonella Infantis and a sanitation indicator Enterococcus faecalis in composts at different stages of the composting process and during storage. Methods and Results: The studied pathogenic and indicator strains, originally isolated from compost, were inoculated into compost samples from the various stages of the composting process. During incubation, indigenous microflora diversity was monitored with DGGE analysis. After 90 days of incubation, strain survival was observed in compost sampled before the beginning of the cooling phase, and DGGE analysis demonstrated an increase of microbial diversity up to the cooling phase. However, inoculated strains were not detected in composts after 30, 60 or 90 days of incubation in compost sampled after the start of the cooling phase. Microbial diversity also became stable, and DGGE profiles reached a maximum number of bands at this stage. Conclusions: Strain survival was not observed in stabilized composts. The cooling phase seems to be the turning point for pathogen survival and at this stage the indigenous microflora appeared to play a significant role in suppression. Significance and Impact of the Study: The importance of indigenous microflora in the survival of pathogens in four different composts was demonstrated. Stabilized composts were recommended for spreading on land.     

Application of COMPOCHIP Microarray to Investigate the Bacterial Communities of Different Composts

A microarray spotted with 369 different 16S rRNA gene probes specific to microorganisms involved in the degradation process of organic waste during composting was developed. The microarray was tested with pure cultures, and of the 30,258 individual probe-target hybridization reactions performed, there were only 188 false positive (0.62%) and 22 false negative signals (0.07%). Labeled target DNA was prepared by polymerase chain reaction amplification of 16S rRNA genes using a Cy5-labeled universal bacterial forward primer and a universal reverse primer. The COMPOCHIP microarray was applied to three different compost types (green compost, manure mix compost, and anaerobic digestate compost) of different maturity (2, 8, and 16 weeks), and differences in the microorganisms in the three compost types and maturity stages were observed. Multivariate analysis showed that the bacterial composition of the three composts was different at the beginning of the composting process and became more similar upon maturation. Certain probes (targeting Sphingobacterium, Actinomyces, Xylella/Xanthomonas/Stenotrophomonas, Microbacterium, Verrucomicrobia, Planctomycetes, Low G + C and Alphaproteobacteria) were more influential in discriminating between different composts. Results from denaturing gradient gel electrophoresis supported those of microarray analysis. This study showed that the COMPOCHIP array is a suitable tool to study bacterial communities in composts.     

Growth potential of faecal bacteria in simulated psychrophilic/mesophilic zones during composting of organic waste

Aim: This study investigated the growth potential of Salmonella serotype Typhimurium and faecal indicator organisms in compost materials and the correlation between bacterial growth potential and the physico‐chemical composition of the compost substrate and temperature. Methods and Results: Survival of Salm. Typhimurium, Enterococcus spp. and total coliforms at 14, 24 and 37°C was determined in material of different degrees of maturity collected from composting plants for household waste and manure. All three micro‐organisms showed the potential for growth in the material from active composts (Solvita index 4) but inactivation generally occurred over time in mature compost material (Solvita index 7–8). Conclusions: Salm. Typhimurium had the potential for growth in psychrophilic/mesophilic (P/M) zones of immature compost material and its growth potential correlated negatively with the maturity of the compost and the temperature within the simulated P/M zone. Significance and Impact of the Study: The risk of pathogen regrowth in P/M zones during organic waste composting further emphasizes the importance of good management practices and of avoiding P/M zones in combination with low compost maturity.