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.     

Effect of matured compost as a bulking and inoculating agent on the microbial community and maturity of cattle manure compost

Cattle manure composts were consecutively manufactured. Compost that reached maturity first was used as a bulking and inoculating agent for subsequent compost production. The microbial community was measured through phospholipid fatty acid analysis. Changes in the content of fatty acid methyl esters derived from phospholipids were similar in all the composts. The diversity index for the fatty acid methyl ester content increased in the secondary-produced compost from the onset of composting. Microbial succession was accelerated using matured compost. The proportion of biomarker fatty acids for gram-positive bacteria also increased in the secondary-produced compost from the early stage of composting. Changes in germination index indicated the maturity stage of the compost. The proportion of biomarker fatty acids for gram-positive bacteria was positively correlated to the germination index, indicating that phospholipid fatty acid analysis is an indicator for evaluating the maturity of cattle manure composts.     

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.     

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.     

Chemical transformation,phytotoxicity and nutrient availability in progressive composting stages of wheat straw

Chemical maturity parameters in addition to plant growth limiting factors have been monitored in the course of a 2-month composting experiment. Wheat straw with 5% dry w horse manure was adjusted to C/N= 45 with urea. The pile was rotated and homogeneous samples were taken every four days. The most intense changes in straw fractions occurred in the first 20 days of composting, as suggested by wet chemical analyses, thermogravimetry and ¹³C NMR spectrometry. Nevertheless, plant response to compost application gave significant changes at between 20–60 days that were not clearly reflected by the above techniques. Glasshouse experiments with a soil treated with compost samples taken at the successive transformation stages suggested no linear correlation between composting time and the potential of compost in improving plant yield. In the samples taken after 20 days in the conditions studied, referred to as postmature composts, the ryegrass yield did not depend on most of the organic matter characteristics, but closely paralleled the concentration of available nitrogen and – to lesser extent – phosphorous in the compost. The probable immobilization of these elements in the course of composting was also suggested by plant response experiments with different doses of compost and the addition or not of mineral solution.     

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.     

Investigating microbial activities in compost using mushroom (Agaricus bisporus) cultivation as an experimental system

Investigations into the dynamic nature of composting environments are necessary to understand and ultimately optimise the complex processes that occur. In this study, various parameters were measured to investigate physical, chemical and biological changes that occur in compost during the production of Agaricus bisporus. In addition to monitoring the compost samples during mushroom cultivation, uninoculated samples were maintained for comparative purposes. Principal components analysis of the variables measured showed a clear distinction between the thermophilic Phase I composts, uninoculated Phase II composts and mushroom inoculated composts. Leucine assimilation, a novel technique to composting environments, is presented as suitable method for assessing microbial activity in such systems. Strict agreement between leucine assimilation and fluorescein diacetate (FDA) hydrolysis, a rarely used technique in composting environments, was not observed, suggesting that neither should be used as a sole measure of microbial activity in compost. Association of FDA hydrolysis with the culturable heterotrophic count suggests that FDA hydrolysis may indicate bacterial as opposed to total microbial activity.     

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.     

Composting of de-inking paper sludge with poultry manure at three nitrogen levels using mechanical turning: behavior of physico-chemical parameters.

De-inking paper sludge (DPS) is rich in carbon (C) but poor in nitrogen (N). Thus, it has a high C:N ratio which limits the composting process. Accordingly, the goal of this study was to investigate the effect of three N treatments on DPS composting. Compost piles of 100 m3 were formed by mixing raw DPS with poultry manure and chicken broiler floor litter, giving on average 0.6%, 0.7% and 0.9% total N. The changes in physico-chemical parameters, total weight and fiber losses, and maturity of composting piles of DPS were monitored during 24 weeks. The compost piles had a neutral to alkaline pH throughout the study. Inorganic N decreased whereas organic N increased over time for all treatments. These changes in magnitude were different among N treatments resulting in a final total N content of 0.9% for the 0.6% N treatment whereas final total N contents of 0.7% and 0.9% N were measured for the 0.7% and 0.9% N treatments. The total weight, cellulose and hemicellulose losses were higher in 0.6% N treatment giving the lowest C:N ratio after 24 weeks of composting. However, none of the 24 week-old composts of DPS were mature based on their final C:N ratio and colorimetric test of maturity. Except for copper, their final total trace element contents meet most known standards or guidelines for organic soil conditioners. Overall, 0.6% N treatment was the best to enhance DPS composting using mechanical turning, but a period of more than 24 weeks was required to reach compost maturity.     

Effect of the raw materials and mixing ratio of composted wastes on the dynamic of organic matter stabilization and nitrogen availability in composts of Sub-Saharan Africa

The effect of raw materials and their proportions in initial mixtures on organic matter (OM) stabilization and nitrogen (N) availability during pit composting in Sub-Saharan Africa was assessed using biochemical fractionation and laboratory incubations to characterize composts sampled throughout the composting process. Stabilization of OM occurred more rapidly in mixtures with slaughter-house wastes, it was progressive in mixture with household refuses while tree leaves compost remained unstable. Carbon mineralization from compost samples was positively correlated to water soluble and hemicellulose-like organic fractions. Mixtures containing large proportions of household refuses reached the highest stability and total N but available N remained weak. Slaughter-house wastes in the initial mixtures made possible to reach good OM stabilization and the largest N availability. The nature of initial mixing influenced composting parameters, OM stabilization and N availability. It is suggested mixing household refuses and slaughter-house wastes with tree leaves to reach better amending and fertilizer qualities of composts.     

Estimation of maturity of compost from food wastes and agro-residues by multiple regression analysis

The composting process of food wastes and tree cuttings was examined on four composting types composed from two kinds of systems and added mixture of microorganisms. The time courses of 32 parameters in each composting type were observed. The efficient composting system was found to be the static aerated reactor system in comparison with the turning pile one. Using the multiple regression analysis of all the data (159 samples) obtained from this study, some parameters were selected to predict the germination index (GI) value, which was adopted as a marker of compost maturity. For example, using the regression model generated from pH, NH(4)(+) concentration, acid phosphatase activity, and esterase activity of water extracts of the compost, GI value was expressed by the multi-linear regression equation (p<0.0001). High correlations between the measured GI value and the predicted one were made in each type of compost. As a result of these observations, the compost maturity might be predicted by only sensing of the water extract at the composting site without any requirements for a large-size equipment and skill, and this prediction system could contribute to the production of a stable compost in wide-spread use for the recycling market.     

Effects of the maturity of wood waste compost on the structural features of humic acids

An analytical scheme for the separation of humic substances (HSs) and non-humic substances (non-HSs) was established to estimate the humification index (HI), which was defined as the ratio of HS carbon content to non-HS carbon content. The alkaline compost-extract contained a mixture of HSs and non-HSs, while acidification of the compost-extract resulted in precipitation of humic acid (HA). The acidified supernatant contained fulvic acid (FA) and non-HSs. In the present study, DAX-8 resin was used to separate FA and non-HSs. HI values, which were estimated to evaluate the maturity of wood waste compost, increased with composting duration. To determine the effects of compost maturity on HA structural features, correlations between HI and indicators of the degree of HA humification (atomic ratios, acidic functional group contents, spectroscopic parameters and molecular weight) were investigated. HI values were significantly related to the indicators of the extent of HA humification during composting.     

Comparison between UV spectroscopy and Nirs to assess humification process during sewage sludge and green wastes co-composting

The humification of organic matter during composting was studied by the quantification and monitoring of the evolution of humic substances (Humic Acid-HA and Fulvic Acid-FA) by UV spectra deconvolution (UVSD) and near-infrared reflectance spectroscopy (NIRS) methods. The final aim of this work was to compare UVSD to NIRS method, already applied on the same compost samples in previous studies. Finally, UVSD predictions were good for HA and HA/FA (r² of 0.828 and 0.531) but very bad for FA (r² of 0.092). In contrary, all NIRS correlations were accurate and significant with r² of 0.817, 0.806 and 0.864 for HA, FA and HA/FA ratio respectively. From these results, HA/FA ratio being a well-used index of compost maturity, UVSD and NIRS represent two invaluable tools for the monitoring of the composting process. However, we can note that NIRS predictions were more accurate than UVSD calibrations.     

Roles of composts in soil based on the assessment of humification degree of fulvic acids

This study was conducted to assess the humification degree in fulvic acids (FA) from different composts, and to reveal their roles after soil amending based on their excitation-emission matrices (EEM) of the fluorescence spectra and projection pursuit regression (PPR) analysis. Two peaks were detected in EEM spectra of FA from all composts, and three components were identified by the parallel factor analysis (PARAFAC) model. The assessment of the humification degree of FA using the ratios between the values of the percent fluorescence response in the visible and ultraviolet regions (P_I,n/P_II,n) generally agreed with that using the distributions of FA components (C1, C2 and C3). The PPR considering the parameters (P_I,n/P_II,n, C1, C2 and C3) further ranked the composts with similar of FA, and the FA humification degree decreased in the order: GW, TSW, LW and SW > CM, MSW, and PM > MC and KW. The results showed that the compositions of FA were similar to each other in composts from different materials, but there were distinct differences in the humification degree of FA owing to the different distributions of each component in composts. Therefore, based on the redistribution of components, a method for regulating the humification degree of FA during composting was presented. Furthermore, the suitable soil application of composts with different humification degrees was also demonstrated.     

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     

Transformation of organic matter during co-composting of pig manure with sawdust

Co-composting of pig manure with sawdust was studied in order to characterize the organic transformation during the process, using both chemical and spectroscopic methods. Humic acids (HA) and fulvic acids (FA) were fractionated from immature and mature pig manure compost, and characterized. After 63 days of composting, the ratio of total organic carbon and soluble organic carbon decreased to a satisfactory low level and the solid and soluble C/N ratios decreased rapidly for the first 35 days before attaining a constant value, indicating compost maturity. Humification could be responsible for the increase in humic acid proportion during composting. The increase in the aromatic bonds after composting, as indicated by the reduction of C/H and C/O ratios of HA and FA, resulted in a more stabilized product. A substantial increase in high molecular weight compounds along with a small increase in low molecular weight compounds was found in mature compost. Moreover the HA also had more complex organic compounds at this stage. Fluorescence spectral analysis showed an increase in the maximum wavelength of HA associated with the contents of aromatic structures in solution. A decrease in relative absorbance of HA at 1160 cm(-1), 2950 cm(-1) and 2850 cm(-1) was seen in the FTIR spectra indicating the decomposition of complex organic constituents, into simpler ones. Increase in the aromatic compounds with higher stability could account for the relative increase in the absorbance of HA at 1650 cm(-1) and 1250 cm(-1) of the mature compost. The composition of FA was not much altered, indicating most of the degradation of organic matter occurred in HA. Data from organic carbon, C/N ratio, elemental analysis, E(4)/E(6) ratio, gel chromatography, fluorescence and FTIR spectra indicated an increase in polycondensed structures and the presence of more stable organic matter in the mature compost.     

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.     

Evaluation of maturity and stability parameters of composts prepared from agro-industrial wastes

The objective of this study was to evaluate changes in physical, chemical and biological parameters to assess the maturity and stability of composts prepared from mixture of different farm and agro-industrial wastes over a period of 150 days. All the composts appeared granular, dark grey in color without foul odor and attained an ambient temperature at 120 days of composting indicating the stable nature of composts. Correlation analysis showed that the optimal values of the selected parameters for our experimental conditions are as follows: organic matter loss >42%, C:N ratio <15, water soluble organic carbon (C(w)):organic N (N(org)) ratio <0.55, humic acid (HA):fulvic acid (FA) ratio >1.9, humification index (HI) >30%, cation exchange capacity (CEC):total organic carbon (TOC) ratio >1.7 and germination index (GI) >70%. Compost enriched with sewage sludge, pressmud and poultry waste matured earlier compared to composts either enriched with distillery effluent or un-enriched.     

Enhancing the co-composting of olive mill wastes and sewage sludge by the addition of an industrial waste

In this work, the effect of incorporating an acidic ferrous sulphate waste (SF) over co-composting process of sewage sludge and olive mill solid wastes in a 1:2 v/v wet basis was investigated. The SF used was an industrial by-product of titanium oxide synthesis and its addition resulted in a chemical stabilisation of the wastes at low pH. The optimum dose of SF to enhance the composting of the studied biowastes was a 20% v/v (wet basis) and the best moment for the addition turned out to be whenever the composting piles had achieved the thermophilic range. The addition of SF over the composting process made possible a faster stabilisation, increasing the composting rate from 0.033 to 0.13 d(-1), and leading to a Fe and S rich compost. All composts obtained fulfilled the limits determined by current European and Spanish regulations and presented better characteristics for its use as soil amendment and organic fertilizer than the traditional composts without SF. The optimum dose of compost containing SF was determined through agronomic tests being its value about 18 Ton ha(-1).