Changes in fungal population of fly ash and vinasse mixture during vermicomposting by Eudrilus eugeniae and Eisenia fetida: documentation of cellulase isozymes in vermicompost

Fly ash (FA) and vinasse (VN), two industrial wastes, are generated in huge amounts and cause serious hazards to the environment. In this experiment, different proportions of these two wastes were used as food for two epigeic earthworms (Eisenia fetida and Eudrilus eugeniae) to standardize the recycling technique of these two wastes and to study their effect on fungal especially cellulolytic fungal population, cellulase activity and their isozyme pattern, chitin content and microbial biomass of waste mixture during vermicomposting. Increasing VN proportion from 25% to 50% or even higher, counts of both fungi and cellulolytic fungi in waste mixtures were significantly (P ? 0.05) increased during vermicomposting. Higher cellulase activity in treatments having 50% or more vinasse might be attributed to the significantly (P ? 0.05) higher concentration of group I isozyme while concentrations of other isozymes (group II and III) of cellulase were statistically at par. Higher chitin content in vinasse-enriched treatments suggested that fungal biomass and fungi-to-microbial biomass ratio in these treatments were also increased due to vermicomposting. Results revealed that Eudrilus eugeniae and Eisenia fetida had comparable effect on FA and VN mixture during vermicomposting. Periodical analysis of above-mentioned biochemical and microbial properties and nutrient content of final vermicompost samples indicated that equal proportion (1:1, w/w) of FA and VN is probably the optimum composition to obtain best quality vermicompost.     

Changes in microbial properties and nutrient dynamics in bagasse and coir during vermicomposting: Quantification of fungal biomass through ergosterol estimation in vermicompost

In this experiment, different microorganisms viz., Trichoderma viridae, Aspergillus niger and Bacillus megaterium were inoculated in bagasse and coir with the objective to study their effect on nutrient dynamics and microbial properties, specially effect on fungal status in these waste materials. Fungal biomass (FBC) was calculated from the ergosterol content in the vermicompost samples. Inoculation of B. megaterium registered comparatively higher TP content in the final stabilized product. Vermicomposting increased microbial biomass carbon (MBC) and nitrogen (MBN) content in bagasse and coir. Microbial biomass carbon to nitrogen ratio (MBC/ MBN) was comparatively narrower in fungi inoculated vermicomposts and FBC/MBC ratio was increased up to 11.69 from 9.51 of control during vermicomposting.     

Microbial biomass in a soil amended with different types of organic wastes.

Application of different types of organic wastes may have a marked effect on soil microbial biomass and its activity. The objective of this study was to quantify the amount of microbial biomass in a loamy-clayey soil, amended with different types of organic waste residues (composts of municipal solid waste of different ages, sewage sludge and farmyard manure) and incubated for 8 weeks at 25 degrees C and two-thirds of field capacity, using the fumigation-extraction method. Both microbial biomass-C and -N (BC and BN, respectively) appeared to be dependent on the type of organic waste residues, on their degree of stability, and on their chemical characteristics. In general, organic wastes increased the microbial biomass-C content in the soil and the microbial BC was positively correlated with the organic C content, the C/N, neutral detergent fibre/N (NDF/N) and acid detergent fibre/N (ADF/ N) ratios. The microbial biomass content decreased according to the period of incubation, especially when the compost used was immature. The microbial biomass-N was positively correlated with the total N and percentage of hemicellulose. The microbial biomass-C was linearly related with the microbial biomass-N and the ratio BC/BN was exponentially related with the BC.     

Microbial inoculants for small scale composting of putrescible kitchen wastes

This research looked at the need for ligno-cellulolytic inoculants (EM bacteria and Trichoderma sp.) in small to medium scale composting of household wastes. A mixture of household organic waste comprised of kitchen waste, paper, grass clippings and composted material was subjected to various durations of thermo composting followed by vermicomposting with and without microbial inoculants for a total of 28 days. The results revealed that ligno-celluloytic inoculants are not essential to speed up the process of composting for onsite small scale household organic waste treatment as no significant difference was observed between the control and those inoculated with Trichoderma and EM in terms of C:N ratio of the final product. However, it was observed that EM inoculation enhanced reproductive rate of earthworms, and so probably created the best environment for vermicomposting, in all treatment groups.     

Land application of biosolids. Soil response to different stabilization degree of the treated organic matter

The effect of land application of biosolids on an agricultural soil was studied in a 2-month incubation experiment. The soil microbial biomass and the availability of heavy metals in the soil was monitored after the application of four different composting mixtures of sewage sludge and cotton waste, at different stages of composting. Land application caused an increase of both size and activity of soil microbial biomass that was related to the stabilization degree of the composting mixture. Sewage sludge stabilization through composting reduced the perturbance of the soil microbial biomass. At the end of the experiment, the size and the activity of the soil microbial biomass following the addition of untreated sewage sludge were twice those developed with mature compost. For the mature compost, the soil microbial biomass recovered its original equilibrium status (defined as the specific respiration activity, qCO2) after 18 days of incubation, whereas the soil amended with less stabilized materials did not recover equilibrium even after the two-month incubation period. The stabilization degree of the added materials did not affect the availability of Zn, Ni, Pb, Cu, Cr and Cd in the soil in the low heavy metal content of the sewage sludge studied. Stabilization of organic wastes before soil application is advisable for the lower perturbation of soil equilibria status and the more efficient C mineralization.     

Potential emissions from two mechanically-biologically pretreated (MBT) wastes

The interaction of parameters determining the potential emissions of two different mechanically-biologically pretreated municipal solid wastes (MBT wastes) is elucidated in this work. The origins of the wastes are Germany and Sweden. By means of lab-scale experiments, increased stabilisation through composting is preferably determined by a decrease in respiration activity. Concurrently, the stabilisation is verified for the leachates by a decrease in COD, DOC, and BOD(5). Total organic carbon content reflects stabilisation less accurately. FT-IR and thermal analytical methods add valuable information about the state of degradation, especially when several distinct thermal parameters are taken into account. Mobility of Cr, Ni, Pb, and Zn produced by a batch leaching test with deionized water is reduced by the pretreatment of both materials. Mobility of copper unambiguously increased. A principle component analysis (PCA) of membrane fractionated leachates indicates an affinity of Cu to mobile humic acids or dissolved organic carbon. High Cr, Zn, and Ni contents in the solid co-occur with high contents of solid humic acids. To a lesser extent, this is also true for solid Cd, Cu, and Pb contents. Due to differences in required landfilling conditions, actual emissions and after-care phase length will depend on whether each waste is landfilled in Germany or Sweden.     

Sludge earthworm composting technology by Eisenia fetida

Prior to vermicompost application to the soils, there is a need to determine the heavy metal concentrations in the final vermicomposts. Acute toxicity on Eisenia fetida for copper (Cu) and cadmium (Cd) were conducted by artificial soils tests. The dosage of interest is typically the median lethal concentration (LC₅₀) that will kill 50 % of the population of organisms within the test period. The dry artificial soil is pre-moistened 1 or 2 days before the test by adding deionized water to obtain approximately half of the required final water content of 40–60 % of the maximum water holding capacity. The 14-day LC₅₀ values (95 % confidence interval) for Cu and Cd were 530 (450–624) and 1118 (988–1265) mg kg^?1, respectively. The vermicomposting was designed to evaluate the effects of earthworm activity on heavy metals in sewage sludge. Compared with the sludge before vermicomposting by Eisenia fetida, the results are as follows: (1) water content, the pH value and organic matter content decreased, (2) total nitrogen content increased, total phosphorus content and total potassium content decreased, (3) available nitrogen concentration, available phosphorus content increased, and (4) the total content of five metals (Cu, Ni, Cd, Pb and Zn) decreased, and the bioaccumulation factor shows that vermicomposting can efficiently remove heavy metals. Therefore, it can be concluded that the soil use of sludge of the Wastewater Treatment Plant in Huaibei is feasible.     

Study of the organic matter evolution during municipal solid waste composting aimed at identifying suitable parameters for the evaluation of compost maturity

In this work the composting process of municipal solid wastes was studied in order to characterize the transformations of organic matter, particularly humic acid (HA). A composting process, lasting three months, was monitored by chemical methods; the following parameters were measured: water-soluble carbon concentration (WSC) and humic substances content (humic and fulvic acid (FA)). The effects of humification on the molecular structure of humic acid (HA) were also evaluated by Fourier transform infrared (FT-IR) and (13)C NMR spectroscopy. WSC concentration rapidly increased reaching a maximum at day-14 of the composting process and then declined. The humic and fulvic acid content (HA and FA, respectively) slightly increased during the process. The FT-IR and (13)C NMR spectra of HA indicate a high rate of change in structure during composting. The groups containing aromatic and carboxylic C increased, while polysaccharides and other aliphatic structures degraded during composting, resulting in HA structures of higher aromaticity. Therefore, spectrometric measurements could provide information significantly correlated to conventional chemical parameters of compost maturity.     

Impact of DOM from composted "alperujo" on soil structure, AM fungi, microbial activity and growth of Medicago sativa

Water-soluble extracts from compost may represent an alternative nutrient and organic matter source for crop production under drip irrigation. Dissolved organic matter (DOM), extracted from composted "alperujo", the main by-product from the Spanish olive oil industry, was applied to soil alone or in combination with either Glomus intraradices Schenck and Smith or a mixture of G. intraradices, Glomus deserticola (Trappe, Bloss. and Menge) and Glomus mosseae (Nicol and Gerd.) Gerd. and Trappe. Response measurements included mycorrhizal colonisation, nutrient uptake and growth of Medicago sativa and microbiological and physical properties in the rhizosphere. Dissolved organic matter was added to soil at concentrations of 0, 50, 100 or 300mgCkg(-1) substrate. During the four months of the experiment, the plants were harvested three times. Both mycorrhizal inoculation treatments significantly increased soil aggregate stability. Only the mycorrhizal inoculations increased microbial biomass C and protease and phosphatase activities and decreased water-soluble C, particularly the mixture of arbuscular mycorrhizal fungi. At the third harvest, the greatest increase in growth of M. sativa was observed in the inoculated plants with shoot biomass being 38% greater than for plants grown in the soil amended with the highest dose of DOM and 57% greater than for control plants. The addition of DOM was not sufficient to restore soil structure and microbial activity and did not affect the mycorrhizal development of introduced populations of arbuscular mycorrhizal fungi, but, depending on the dose, its fertiliser efficiency for improving plant growth was apparent.     

Changes in enzymatic activities and microbial properties in vermicompost of water hyacinth as affected by pre-composting and fungal inoculation: A comparative study of ergosterol and chitin for estimating fungal biomass

In this experiment, three different fungal species, viz. Trichoderma viridae, Aspergillus niger and Phanerochaete chrysosporium, were inoculated in 7 day and 15 day partially decomposed water hyacinth to study their effect on enzymatic activities, microbial respiration and fungal biomass of the final stabilized product. The results suggested that increasing the duration of pre-composting from 7 days to 15 days did not show any significant effect on the activities of hydrolytic enzymes. Inoculation of fungi significantly (P ? 0.05) increased cellulase, protease and acid and alkaline phosphatase activities. The highest value of ergosterol was recorded in A. niger-inoculated vermicomposts. Inoculation of P. chrysosporium in initial organic waste registered the highest chitin content in vermicompost. A comparison of fungal biomass and chitin content revealed a conversion factor of 2.628 with a standard deviation of 0.318. Due to significant correlation (r = 0.864), this conversion factor allows for the calculation of fungal biomass from chitin, which is comparatively more stable than ergosterol.     

A full-scale study of treatment of pig slurry by composting: kinetic changes in chemical and microbial properties

Since the indiscriminate disposal of pig slurry can cause not only air pollution and bad odours but also nutrient pollution of ground waters and superficial waters, composting is sometimes used as one environmentally acceptable method for recycling pig manure. The aim of this study was to evaluate the effect of composting pig slurry on its sanitation (evaluated by ecotoxicity assays and pathogen content determination), as well as to determine the effect of a carbon-rich bulking agent (wood shavings, WS) and the starting C/N ratio on the changes undergone by different chemical (volatile organic matter, C and N fractions) and microbiological (microbial biomass C, ATP, dehydrogenase activity, urease, protease, phosphatase, and beta-glucosidase activities) parameters during composting. Pig slurry mixed with bulking agent (P+WS) and the solid faction separated from it, both with (PSF+WS) and without bulking agent (PSF), were composted for 13 weeks. Samples for analysis were taken from composting piles at the start of the process and at 3, 6, 9, and 13 weeks after the beginning of composting. The total organic carbon, water soluble C and ammonium content decreased with composting, while Kjeldahl N and nitrate content increased. The nitrification process in the PSF+WS pile was more intense than in the PSF or P+WS composting piles. The pathogen content decreased with composting, as did phytotoxic compounds, while the germination index increased with compost age. Piles with bulking agent showed higher values of basal respiration, microbial biomass carbon, ATP and hydrolase activities during the composting process than piles without bulking agent.     

Selective organic compounds degradation under controlling composting conditions

Organic matter stabilization resulted from the decrease of cellulose, xylan, arabinan, acetyl groups, glucuronic acids, galacturonic acids (easily biodegradable fractions) and the increase of lignin (resistant compound) and humic substances coming from the initial wastes have been studied. A central composite experimental design was used to investigate the influence of environmental composting parameters (moisture, aeration and particle size) on organic matter evolution. The organic matter evolution was clearly influenced by the studied composting parameters. All results were concordant, with an increase of humic substances and lignin and a decrease of the rest of the cellulose and hemicellulose compounds. Lower cellulose, xylan, acetyl groups and glucuronic acids contents (higher degradation) have been observed under low particle size (1 cm) and higher moisture content (70%). However lower lignin and higher humic substances under medium (3 cm) to low particle size and low moisture content (40%) have been found.     

CHARACTERIZATION OF URBAN WASTES ACCORDING TO FERTILITY AND PHYTOTOXICITY PARAMETERS

Several urban wastes of different nature and level of organic matter stability (municipal solid wastes, sewage sludges and composts) have been characterized analysing fertility and phytotoxicity parameters. Sewage sludges showed the highest N and P values of all the wastes. The total K content of the wastes was low but almost all was available to plants. Mature composts had the lowest organic carbon and humic substances values as a consequence of organic matter mineralization during the composting process. The soluble C at pH 2/precipitated C ratio decreased as the stability of the organic matter increased. In spite of the heterogeneity of the wastes analysed, the organic matter/total organic C ratio was largely constant with a mean value of 2.05. In no case did the heavy metal levels exceed the maximum allowed by the EU disposition for sewage sludges use in agriculture. The highest levels of phytotoxic substances occurred in the fresh wastes. The germination rates and root length were highest with mature composts. Germination index and root length were negatively correlated with water soluble carbon (WSC) and WSC/N ratio. The principal component analysis showed that extractable C, soluble C at pH 2 and water soluble C were the C fractions which most contributed to the total variability.     

Functional changes in culturable microbial communities during a co-composting process: Carbon source utilization and co-metabolism

Microbial communities in sewage sludge and green waste co-composting were investigated using culture-dependent methods and community level physiological profiles (CLPP) with Biolog Microplate. Different microbial groups characterized each stage of composting. Bacterial densities were high from beginning to end of composting, whereas actinomycete densities increased only after bio-oxidation phase i.e. after 40 days. Fungal populations become particularly high during the last stage of decomposition. Cluster analyses of metabolic profiles revealed a similar separation between two groups of composts at 67 days for bacteria and fungi. Principal component analysis (PCA) applied to bacterial and fungal CLPP data showed a chronological distribution of composts with two phases. The first one (before 67 days), where the composts were characterized by the rapid decomposition of non-humic biodegradable organic matter, was significantly correlated to the decrease of C, C/N, organic matter (OM), fulvic acid (FA), respiration, cellulase, protease, phenoloxidase, alkaline and acid phosphatases activities. The second phase corresponding to the formation of polycondensed humic-like substances was significantly correlated to humic acid (HA) content, pH and HA/FA. The influent substrates selected on both factorial maps showed that microbial communities could adapt their metabolic capacities to the particular environment. The first phase seems to be focused on easily degradable substrate utilization whereas the maturation phase appears as multiple metabolisms, which induce the release of metabolites and their polymerization leading to humification processes.     

Measurement of microbial biomass and activity in landfill soils

Two complementary techniques, which have been widely used to provide a general measure of microbial biomass or microbial activity in natural soils, were evaluated for their applicability to soils from the Mallard North and Mallard Lake Landfills, DuPage County, Illinois, U.S.A. Included were: (1) a potassium sulphate extraction technique with quantification of organic carbon for measurement of microbial biomass; and (2) an arginine ammonification technique for microbial activity. Four profiles consisting of replaced soils were sampled for this study; units included topsoil (mixed mollisol A and B horizons), compacted clay cover (local calcareous Wisconsinan age glacial till), and mixed soil/refuse samples. Internally consistent results across the four profiles and good correlations with other independent indicators of microbial activity (moisture, organic matter content, nitrogen, and phosphorus) suggest that, even though these techniques were developed mainly for natural mineral soils, they are also applicable to disturbed landfill soils.     

Heavy metal and nutrient changes during vermicomposting animal manure spiked with mushroom residues

A pilot-scale trial of four months was conducted to investigate the responses of heavy metal and nutrient to composting animal manure spiked with mushroom residues with and without earthworms. Results showed that earthworm activities accelerated organic matter mineralization (e.g. reduction in C/N ratio, increase in total concentrations of N, P, K) and humification (e.g. increase in humic acid concentration, humification ratio and humification index). Despite composting increased total heavy metal (i.e. As, Pb, Cu, Zn) concentrations irrespective of earthworm, the availability of heavy metals extracted by DTPA significantly (P < 0.05) decreased particularly in treatments with earthworms introduced. The shift from available to unavailable fractions of heavy metals was either due to earthworm bioaccumulation, as indicated by total heavy metal concentrations being higher in earthworm tissues, or due to the formation of stable metal-humus complexes as indicated by the promotion of humification. Our results suggest that vermicomposting process could magnify the nutrient quality but relieve the heavy metals risk of agricultural organic wastes.     

Continuous-feeding vermicomposting as a recycling management method to revalue tomato-fruit wastes from greenhouse crops

Huge quantities of discarded fruits generated from greenhouse crops represent a worldwide environmental problem. The aim of this work was to assess the efficiency of vermicomposting as a recycling management option for biotransforming tomato-fruit wastes from greenhouses into an organic nutrient-rich product available for agricultural purposes. A pilot vermireactor was constructed. It was provided with a manure layer, where an initial population of Eisenia fetida was introduced and fed continuously at a high organic loading rate (13.6 kg TOC m^?3 wk^?1) for 150 days. Vermicompost chemical and enzymatic parameters as well as the bacterial and fungal community structure were determined for 210 days (vermicomposting plus a maturation period). Earthworm biomass increased after 90 days, and then declined due to increasing pH, electrical conductivity and ammonium concentration. The temporal patterns of dehydrogenase, β-glucosidase, protease and urease were related to earthworm growth and the stabilization of organic matter. Bacterial DGGE profiles differed between the period of degradation of labile substrates and the maturation step. Fungal communities at the stage of maximum earthworm biomass differed most, suggesting a gut passage effect. The end product was chemically stable and enriched in nutrients, demonstrating that tomato-fruit wastes can be successfully vermicomposted into a valuable soil amendment. We suggest continuous-feeding vermicomposting as an environmentally sound management option for greenhouse wastes.     

Bioreactor treatment of municipal solid waste landfill leachates: characterization of organic fractions

Quantitative and qualitative changes in organic matter were studied at different stages of treatment in a bioreactor designed to process leachates from a municipal solid waste landfill. The particulate matter (PM) and macromolecular fractions of the dissolved organic matter with solubility properties comparable to humic (acid-insoluble) and fulvic (acid-soluble) acid fractions (AI, AS, respectively) from the incoming black liquid, the bioreactor content, and the final processed effluent were isolated, quantified, and characterized by visible and infrared (IR) spectroscopies. The macromolecular signature either aliphatic (glycopeptides, carbohydrates) or aromatic (coinciding with infrared patterns of lignin, tannins etc.) enabled us to characterize the different organic fractions during the course of microbial transformation. The results reveal significant changes in the nitrogen speciation patterns within the different organic fractions isolated from the wastewater. The final increase in the relative proportions of nitrogen in the least aromatic AS fraction during microbial transformation could be related to protein formation inside the bioreactor. After biological treatment and ultrafiltration, the amount of organic matter was reduced by approximately 70%, whereas aromaticity increased in all fractions, indicating preferential elimination of aliphatic wastewater compounds. Most of the remaining fractions at the end of the process consisted of a yellow residue rich in low molecular weight AS fractions.     

Solid-phase fluorescence spectroscopy to characterize organic wastes

The production of solid organic waste (SOW) such as sewage sludge (SS) or municipal solid waste (MSW) has been continuously increasing in Europe since the beginning of the 1990'. Today, the European Union encourages the stabilization of these wastes using biologic processes such as anaerobic digestion and/or composting to produce bio-energy and organic fertilizers. However, the design and management of such biologic processes require knowledge about the quantity and quality of the organic matter (OM) contained in the SOW. The current methods to characterize SOW are tedious, time-consuming and often insufficiently informative. In this paper, we assess the potential of solid-phase fluorescence (SPF) spectroscopy to quickly provide a relevant characterization of SOW. First, we tested well known model compounds (tryptophan, bovine serum albumin, lignin and humic acid) and biologic matrix (Escherichia coli) in three dimensional solid-phase fluorescence (3D-SPF) spectroscopy. We recorded fluorescence spectra from proteinaceous samples but we could not record the fluorescence emitted by lignin and humic acid powders. For SOW samples, fluorescence spectra were successfully recorded for MSW and most of its sub-components (foods, cardboard) but impossible for SS, sludge compost (SC) and ligno-cellulosic wastes. Based on visual observations and additional assays, we concluded that the presence of highly light-absorptive chemical structures in such dark-colored samples was responsible for this limitation. For such samples, i.e. lignin, humic acid, SS, SC and ligno-cellulosic wastes, we show that laser induced fluorescence (LIF) spectroscopy enables the acquisition of 2D fluorescence spectra.     

Comprehensive quality assessment of municipal organic waste composts produced by different preparation methods

In the first part of this work, the effect of municipal organic waste (MOW) composts on plant growth was evaluated in a greenhouse trial. The treatments included soil amended with 14 different composts (prepared by shredding, adding wood shavings, cocomposting with biosolids or vermicomposting), an inorganically fertilized soil, and a control soil. All of the treatments significantly increased plant growth compared to the control, and yields of three of the amended treatments were as high as that of the inorganic fertilizer treatment. When comparing differently prepared composts to the conventional compost, it was found that cocomposting MOW with biosolids was the method which most positively influenced yields (26-41% yield increases). In the second part of this work, we evaluated the effects of the different preparation methods on compost quality, using a multivariate approach. Three main quality aspects were considered collectively in a principal component analysis: organic matter and nutrient concentrations, degradability and capacity to mineralize these nutrients, and plant growth. The model was restricted to the first and second components (PC1 and PC1) which accounted for 94% of data variance. On the resulting factorial plane, four groups were distinguished. Each of the groups was compared to the reference compost to determine quality increases or decreases. Based on this analysis, it was found that cocomposting MOW with biosolids produced the highest quality products (higher total nutrient and OM concentration, nutrient mineralization potential, and plant growth). Addition of wood shavings increased OM concentration, but reduced quality in terms of the other aspects studied. Shredding was only effective to increase product quality when it was not combined with other methods, whereas vermicomposting only increased quality when MOW was not mixed with biosolids.