Lignocellulose degradation in mushroom composts

The edible mushroomAgaricus bisporus is grown commercially on composted manure/straw mixtures. However, this proven composting procedure is wasteful of raw materials. A nonmanure compost was developed (Smith, 1980) with two main aims:

1. To conserve raw materials, while still producing a compost favoringAgaricus bisporus colonization and giving an economic yield of mushrooms.
2. To speed up composting, hence making more efficient use of labor, farm equipment, and buildings.

A “conservation compost” (wheat straw, bran, whey, urea, peat, and gypsum) is ready for inoculation with mushroom mycelium (spawning) after 7 d preparation, i.e., 2 d pre-wetting of straw, then 4–5 d composting under controlled conditions. Whereas a traditional manure/wheat straw compost is produced by composting in windrows (8–11 d) followed by a controlled pasteurization phase (5–7 d). In the preparation of a traditional mushroom compost, as much as 60% of the initial dry matter is lost by microbial degradation prior to spawning. By shortening the composting process to 7 d conservation of cellulose and hemicellulose is achieved with only some 30% loss in dry matter. Straw hemicelluloses are degraded much quicker than cellulose during composting. Hence, the measurable extracellular laminarinase and xylanase activities of the compost microflora appear much greater than their cellulase activities at this period in both composts. A peak in laminarinase and xylanase activity after 48 h in manure compost corresponds with the increase in microbial populations. A pronounced increase in thermophilic bacterial and actinomycete populations occurs in “conservation composts” as readily available soluble carbohydrates are assimilated. Initially, this results in higher uniform compost temperatures (60?C+) and leads to a reduced thermophilic fungal population (10³ viable propagules g^-1 dry wt compost), which may explain the lowered enzyme activities found in the “conservation composts” and thus the reduced degradation of lignocellulose. The compost microflora showed no laccase activity during composting, and little if any lignin was degraded. However,Agaricus bisporus does possess a moderately active lignolytic system and a strongly active cellulolytic system. Subsequent experiments have shown that increased mushroom yields may be obtained from these composts when urea is replaced by chicken manure as the nitrogen supplement (Smith, 1983); this has not affected compost “selectivity” for mushroom growth, dry matter loss, or the duration of the process. Although yield of mushrooms, based on compost weights at spawning tend to be lower than what would be expected from traditional composts, yield calculated on the basis of weight of starting materials is usually much higher.     

An electrochemical on-field sensor system for the detection of compost maturity

A maturity sensor system was developed, based on the combination of three electrically measured parameters, pH, NH₄⁺ concentration, and phosphatase activity in the water extracts of compost samples. One of these parameters, the apparent phosphatase activity in crude test solutions was determined using screen-printed carbon strips (SPCSs) coated with α-naphthyl phosphate (α-NP) in Nafion film. The phosphatase activity was monitored in connection with differential pulse voltammetry (DPV) with an aliquot (30 μL) of the test solution on SPCS. The phosphatase activity sensor was validated using alkaline phosphatase (ALP) in Tris-HCl buffer (pH 8.0) and acid phosphatase (ACP) in citric acid buffer (pH 5.0). The activity of the spiked enzymes in the water extract of the compost sample could be confirmed with the change of corresponding oxidation peak current signal of the product, α-naphthol. The water extracts of compost samples (n = 24) collected in various composting days were applied to our compost maturity sensor system, and the conventional germination tests. Using multiple regression analysis, the germination index (GI) was expressed by the multi-linear regression equation consisting of pH, NH₄⁺ concentration, and the phosphatase activity. The calculated GI from the regression equation had a good correlation with the measured GI of the corresponding composts (r = 0.873). As a result, we have determined an equation for the determination of the compost stability using our portable sensor system rapidly at the composting site.     

Thermal Drying of <Emphasis Type="BoldItalic">Lactobacillus delbrueckii</Emphasis> subsp. <Emphasis Type="BoldItalic">bulgaricus</Emphasis> and its Efficient Use as Starter for Whey Fermentation and Unsalted Cheese Making

Lactobacillus bulgaricus grown on whey was dried by a simple thermal drying method at the range 35–55°C and its efficiency for lactic acid fermentation of whey was evaluated. Drying of cells in whey suspension in the examined temperature range did not affect significantly their viability (82–87% survival), indicating a protective effect of whey as both growth and drying medium. The kinetics of fermentation of whey and mixtures of whey/molasses using the dried culture were comparable to those of non-dried cells, and only low pH had a detrimental effect on the fermentation ability of the dried cells. Furthermore, dried L. bulgaricus, free or immobilized on casein coagulates, was used as starter for the production of unsalted hard-type cheese. The effects of the amount of starter culture and the immobilization technique, the evolution of microbial counts, and the sensory properties of the produced cheeses were evaluated during ripening at various temperatures.     

Biodegradation and hydrolysis rate of aliphatic aromatic polyester

The biodegradation and hydrolysis rates of an aliphatic aromatic copolyester were measured in manure, food, and yard compost environments and in phosphate buffer solution (pH = 8.0) and vermiculite at 58 °C. Mineralization, molecular weight reduction, and structural changes determined by DSC, FTIR, and ¹H NMR were used as indicators of the biodegradation and hydrolysis rates. Poly(butylene adipate-co-terephthalate), PBAT, film biodegraded at distinctive rates in manure, food, and yard compost environments having different microbial activities. The highest biodegradation rate was found in manure compost, which had the highest CO₂ emissions and lowest C/N ratio. The possible presence of extracellular enzymes in manure and food composts may facilitate the hydrolytic reaction since greater molecular weight reduction rates were observed in these composts. ¹H NMR and thermal analysis revealed that, while PBAT is a semi-crystalline copolyester with cocrystallization of BT and BA dimers, the soft aliphatic domain (BA) and the amorphous region are more susceptible to hydrolysis and biodegradation than the rigid aromatic domain (BT) and the crystalline region.     

A morphological study of mullite long fiber prepared using polyvinyl butyral as spinning aids

Morphologies of aluminosilicate gel fiber and mullite long fiber prepared from spinning sol of Al(NO₃)₃·9H₂O, tetraethylorthosilicate and ethanol using polyvinyl butyral as a spinning aids have been studied experimentally. The weight loss of fibers as function of drying time and calcination temperature was summarized and the fiber morphologies at different temperature were also discussed. The results indicated that fiber sample lost the main weight at the first several minutes at 40°C. A main axial crack was observed due to the heat stress if gel fiber was not pre-dried before calcination. Accompanied by weight loss during heat treatment, “black” fibers were shown at the temperature range of 300–500°C and some attachment was observed among fiber surface attributed by the decomposition of organic materials. The actual elimination temperature of organic materials was different from our former heat analysis measurement result, and it could be explained by the pre-treating procedure and high heating rate of heat analysis. The morphological study of mullite fiber calcined at different temperature could help to understand and obtain the mullite fiber with smooth surface.     

Inactivation of Botrytis cinerea during thermophilic composting of greenhouse tomato plant residues

The effectiveness of in-vessel thermophilic composting on the inactivation of Botrytis cinerea was evaluated. The bioreactor operated on an infected mixture of tomato plant residues, wood shavings, and municipal solid compost (1∶1.5∶0.28). Tap water and urea were added to adjust the moisture content and C∶N ratio to 60% and 30∶1, respectively. Used cooking oil was added as a bioavailable carbon source to compensate for heat losses from the system and extend the thermophilic compositing stage. The controlled thermophilic composting process was successful in inactivating B. cinerea. During all experiments, the average reactor temperature increased gradually, reaching its peak after 31 h of operation. Temperatures in the range of 62.6–63.9°C were maintained during the thermophilic stage by the intermittent addition of used cooking oil. The results of the enzyme-linked immunosorbent assay test indicated that the initial concentration of B. cinerea in the compost samples (14.6 μg of dried mycelium/g of compost) was reduced to 12.9, 8.8, and 2.4 μg/g after 24, 48, and 72 h of thermophilic composting, respectively. Plating assay indicated that the mold was completely inactivated in samples after 48 h of thermophilic composting. No significant reduction in B. cinerea was observed during the transient phase (first 30 h of rising temperature) because the temperature reached the lethal level of 55°C after 23 h, thus allowing only 7 h of exposure to temperatures higher than 55°C during this phase. The relatively short time required for complete inactivation of B. cinerea was achieved by maintaining a constant high temperature and a uniform distribution of temperature and extending the duration of the thermophilic stage by the addition of the proper amount of bioavailable carbon (used cooking oil).     

Biodegradation of packaging materials: composting of polyolefins

The compostability of LDPE, PP and heterophasic E-P Copolymers was studied for 5 months under normal and accelerated composting environments. Bio-susceptibility of pre-UV (290 nm) treated films (∼ 100μm, 5 X 5 cm) was measured by monitoring the weight loss, intrinsic viscosity [η], chain scission, functional group evolution (FT-IR) and surface morphology (SEM). It was found that with the increasing time of UV treatment, weight loss was increased in compost. Almost linear decrease in [η] was observed for irradiated and composted samples. The temperature of compost and extra addition of thermophilic microbes significantly influenced the biodegradation. In general, it was concluded that the composition of copolymer markedly affected the compostability and increased ethylene content, slowed down the microbial activity.     

Characterization of Compost with Respect to its Content of Heavy Metals. Part II: Sample Preparation

Abstract

This paper forms part of a study aimed at the development of a procedure for the determination of heavy metals in compost. The present paper deals with the sample preparation stage, including subdividing laboratory samples and reduction of the particle size.

The effect of particle size reduction on the random error due to the analysis of test portions was examined. It was found that milling for 20min in a quartz planet ball mill yields sufficiently homogeneous test samples. As the milling capacity of this device is too small, prior to milling subdivision of the laboratory sample is required. Subdivision before drying is recommended for practical reasons.     

The influence of biotic and abiotic factors on the rate of degradation of poly(lactic) acid (PLA) coupons buried in compost and soil

Poly(lactic) acid (PLA) is a compostable biopolymer and has been commercialised for the for the manufacture of short-shelf life products. As a result, increasing amounts of PLA are entering waste management systems and the environment; however, the degradation mechanism is unclear. While hydrolysis of the polymer occurs abiotically at elevated temperature in the presence of water, potential catalytic role for microbes in this process is yet to be established. In this study, we examined the degradation of PLA coupons from commercial packaging at a range of temperatures (25°, 37°, 45°, 50° and 55 °C) in soil and compost and compared with the degradation rates in sterile aqueous conditions by measuring loss of tensile strength and molecular weight (Mw). In addition, in order to assess the possible influence of abiotic soluble factors in compost and soil on degradation of PLA, degradation rates in microorganism-rich compost and soil were compared with sterile compost and soil extract at 50 °C. Temperature was determined to be the key parameter in PLA degradation and degradation rates in microorganism-rich compost and soil were faster than in sterile water at temperatures 45° and 50 °C determined by tensile strength and Mw loss. Furthermore, all tensile strength was lost faster after 30 and 36 days in microorganism-rich compost and soil, respectively, than in sterile compost and soil extract, 57 and 54 days, respectively at 50 °C. Significantly more Mw, 68% and 64%, was lost in compost and soil, respectively than in compost extract, Mw, 53%; and in soil extract, 57%. Therefore, degradation rates were faster in microorganism-rich compost and soil than in sterile compost and soil extract, which contained the abiotic soluble factors of compost and soil at 50 °C. These comparative studies support a direct role for microorganisms in PLA degradation at elevated temperatures in humid environments. No change in tensile strength or Mw was observed either 25° or 37 °C after 1 year suggesting that accumulation of PLA in the environment may cause future pollution issues.     

Reduction of pathogenic bacteria in organic compost using gamma irradiation

Organic compost is a useful fertilizer for organic farming. However, it poses a microbiological hazard to the farm products because most of the composts are originated from excremental matters of domestic animals. In this study, the radiation treatment was performed to improve microbiological safety of organic compost and the effectiveness of gamma irradiation for inactivating Salmonella Typhimurium and Escherichia coli was investigated. The total aerobic and coliform bacteria in the 16 commercial composts were ranged from 10⁵ to 10⁷ CFU/ml and 0 to 10³ CFU/ml, respectively. All coliform bacteria in the composts were eliminated by irradiation at a dose of 3 kGy, while about 10² CFU/ml of the total aerobic bacteria were survived up to 10 kGy. In the artificial inoculation test, the test organisms (inoculated at 10⁷ CFU/g) were eliminated by irradiation at 3 kGy. Approximate D₁₀ values of Salmonella Typhimurium and E. coli in the compost were 0.40 and 0.25 kGy, respectively. In the cultivation test, the test organisms of the compost had transfer a lettuce leaves. The growth pattern of lettuce was not different between irradiated and non-irradiated composts.     

Abiotic and biotic degradation of oxo-biodegradable foamed polystyrene

An alternative for improving the degradability of polyolefins and polystyrene is the addition of pro-oxidant substances to their formulations. The materials obtained are then called oxo-biodegradable. This work aimed to assess the biotic and abiotic degradation of atactic polystyrene (PS), utilising as test material foamed PS plates used in the manufacture of trays, formulated with Co- and Mn-based pro-oxidant additives. The plates were exposed to artificial weathering (ultraviolet radiation and heat) and were periodically analysed for changes in structural properties. The oxidised surface residues detached from the samples were incubated in a stabilised compost of urban waste (58 °C) or in an aqueous mineral medium (25 °C), the latter being inoculated with urban waste compost and also with a microbial consortium. It was found that the molar masses of the eroded materials from the pro-oxidant activated samples were significantly lower than the initial sample molar masses, with simultaneous incorporation of oxygen into the chains during the accelerated weathering. These samples underwent biodegradation and gave mineralisation values of 2-5% over 2-3 months of incubation in compost and perlite or in mineral aqueous medium. Biodegradation of the residues from the samples not containing pro-oxidant additives was also observed, but at levels which were lower than those obtained for oxo-biodegradable samples.     

A kinetic study of trace element leachability from abandoned-mine-polluted soil treated with SS-MSW compost and red mud. Comparison with results from sequential extraction

The effect of adding treated red mud, a by-product of alumina production, to soil polluted by an abandoned mine and characterised by high concentrations of heavy metals, relatively low reaction grade, and low organic carbon content, was investigated. Also studied was addition of both red mud and compost (produced from source-separated municipal solid waste)—the synergistic action of red mud and compost could be exploited to achieve both metal trapping and an increase in organic carbon content. Leaching batch tests were performed on four different systems: soil, soil and treated red mud, soil and compost, soil and compost plus treated red mud. Dilute sulfuric acid and EDTA solution (liquid/solid ratio 10:1) were used in the tests—sulfuric acid to mimic acid rain and EDTA in accordance with general methods for estimating plant-available metals. Sequential extraction was also applied to the same samples. The use of relatively non-specific extractant reagents in the leaching tests led to a kinetic approach (already proposed in literature), because measurements of trace elements extracted at equilibrium cannot be related to their speciation. Comparison of information obtainable by the kinetic approach to evaluation of data from leaching tests with results from sequential extraction enabled evaluation whether the kinetic fractionation method, a relatively rapid and simple procedure, furnishes adequate information about the mobility and bioavailability of trace elements. Especially interesting results were obtained for Mn, Zn, and Ni, present in large amounts in the soil studied—their leachability was significantly reduced by addition of red mud and compost, suggesting interesting perspectives in soil-remediation activity.     

Effect of storage of soil at 4°C on the microbial activity studied by microcalorimetry

The effect of the storage of soils on their microbial activity has been studied using a microcalorimetric method. Soil samples were kept in closed polyethene bags at 4°C during 3–6 months. Results show changes in the slope of the differentP?t curves recorded from the samples stored at 4°C. This fact strongly suggest the existence of changes of the microbial activity of soil as the heat evolution is a direct measurement of the cells metabolic activity. The value of the Peak-time (time in which the microcalorimetric signal reaches the maximum value) is related with the microbial density of soil samples. This parameter was affected by the time of storage increasing with time. The total heat evolutionQ(t), of the soil samples amended with glucose calculated from the area limited by the Power-Time curves, also decreases with the time of storage. The soil that had been stored for 6 months before experiments, showed the lowest valu ofQ(t).     

Controlled composting of straw

Production of composts on the farm from surplus straw might provide a low-cost biotechnological approach for increasing the value of this lignocellulosic waste. Successful composting depends on the conversion of the polysaccharides (cellulose and hemicelluloses) of straw by inoculated microorganisms to products that can promote plant growth when applied to the land. None of the potentially useful products we have identified are produced by cellulolytic organisms. We have therefore studied mixed populations in which noncellulolytic bacteria depend for growth on the products of fungal cellulolysis. The nature and yield of bacterial products depends not only on conditions within the compost, but also on the microbial inoculants used. Under defined laboratory conditions, using pure cellulose, N₂ is fixed by the anaerobic bacteriumClostridium butyricum in association with a cellulolytic fungus such asTrichoderma sp. A similar association has been achieved on straw withPenicillium corylophilum as the cellulolytic fungus. Cellulolytic fungi can also provide available substrates for the production of bacterial polysaccharides that can improve the structure of unstable soils. The yield of polysaccharide and its efficacy in soil aggregate stabilization again varies with the inoculants used. Such composts can thus contribute to plant nutrition and to soil structure. The adoption ofTrichoderma spp. as the cellulolytic inoculants would further extend the potential value of the compost to include the biocontrol of plant pathogens.     

Evaluation of the aerobic composting process of winery and distillery residues by thermal methods

The possibility of using thermal analysis for a quick characterization of chemical changes was tested in the organic matter from composting materials. Differential thermal analysis (DTA), thermogravimetry (TG) and the first derivative of the TG (DTG) were calculated in oxidizing conditions on compost samples obtained from three composting piles. The composting piles were made by mixing winery and distillery residues with sewage sludge (pile 1), with cow manure (pile 2) and hen droppings (pile 3). The temperature values in the pile 1 showed a different evolution during the thermophilic stage of the composting process in relation to the piles 2 and 3. The thermophilic stage for pile 1 was 17 days, meanwhile for the piles 2 and 3 were around 80 and 110 days, respectively, and probably pile 1 was not well composted. The curves of ion current of CO₂ have been recorded in order to shed light on changes occurring in organic matter during composting. Particularly DTG curves allowed us to distinguish between well (piles 2 and 3) and poor (pile 1) stabilized organic matter. The energy released was calculated for each sample by integrating the DTA curves and these results are agreed with the previous hypothesis. Information deriving from weight losses, registered by the TG and DTG curves, enables to follow the evolution state of the organic matter and therefore changes in its stability. These data could determine the final point of the composting process of winery and distillery residues and then reduce the time for compost harvest.     

Biodegradability of bio-flour filled biodegradable poly(butylene succinate) bio-composites in natural and compost soil

This study investigated the biodegradability of PBS and bio-flour, which is a poly(butylene succinate) (PBS) bio-composite filled with rice-husk flour (RHF) reinforcing, in natural and aerobic compost soil. The percentage weight loss and the reduction in mechanical properties of PBS and the bio-composites in the compost soil burial test were significantly greater than those in the natural soil burial test. These results were supported by degraded surface of PBS and bio-composites observed through morphological study and the total colony count of natural soil was lower than that of compost soil. The biodegradability of the bio-composites was enhanced with increasing bio-flour content because the bio-flour is easily attacked by microorganisms. As the biodegradability test progressed over time up to 80 days, the molecular weight of PBS decreased in the soil burial test. We confirmed by attenuated total reflectance (FTIR-ATR) analyser that the chemical structures of PBS and the bio-composites were changed after the compost burial test. The glass transition temperature (T_g), melting temperature (T_m), crystallization temperature (T_c), heat of fusion (ΔH_f) and heat of crystallization (ΔH_c) of the natural and composted soil tested PBS were investigated using differential scanning calorimetry (DSC). From the results, we concluded that use of these bio-composites will reduce the environmental problems associated with waste pollution and the study findings support the predicted application of bio-composites as “green-composites” or “eco-materials”.     

Development of an experimental procedure to analyse the ‘soil health state’ by microcalorimetry

This work is a ‘historical’revision of the evolution of an experimental procedure developed by Prof.Lisardo Núñez and his research group TERBIPROMAT to study thesustainability and the soil health state.From the very beginning,in 1993, the microbial activity was the main bioindicator selected to analysethe ‘soil health state’. For this reason, a microcalorimetrictechnique was used lately to analyse the influence of different human activitiessuch as reforestations, agricultural exploitation or pollution on the microbialactivity in different soils. Microcalorimetry is the main scientific techniqueused in this research to follow the stimulation of the microbial activityby addition of glucose. The data obtained were complemented by a study ofphysical, chemical and biological parameters of soil and allowed to followthe microbial activity in soils of Galicia (Spain) along the year.The final results, still in revision, will be helpful in establishinga data basis for real maps of the ‘health state’ of differentsoils. Such maps could be used to design processes that help us to decidehow we should exploit soils ensuring their sustainability.     

Quantitative determination of organic priority pollutants in the composts of sewage sludge with rice straw by gas chromatography coupled with mass spectrometry

In order to investigate the occurrence and distribution of organic contaminants in the compost of sewage sludge with rice straw, four different composting treatments at a low carbon/nitrogen (C/N) ratio (13:1) were conducted. Thirty semi-volatile organic compounds (SVOCs) listed as priority pollutants by both China and the US Environmental Protection Agency (EPA) were Soxhlet-extracted, separated and analyzed by gas chromatography coupled with mass spectrometry (GC-MS). The results showed that after composting (56 days) most of SVOCs were detected in the final composts. The total concentrations of polycyclic aromatic hydrocarbons (PAHs) and phthalic acid esters ranged from 1.8 to 10 mg/kg dry weight (d.w.) and from 9.8 to 18 mg/kg d.w., respectively, being significantly higher than those of chlorobenzenes and nitroaromatic compounds which were generally less than 1.0 mg/kg d.w. The concentrations and predominant compounds of organic contaminants in the different composts varied, and were affected by their physico-chemical properties and the composting processes. Concentrations of SVOCs in the static aerated composting processes especially intermittently aerated composting treatment were considerably lower than those in the manual-turned composting treatments. Concentrations of organic contaminants especially PAHs and di(2-ethylhexyl) phthalate in the final composts were in agreement with the maximum permissible limits for agricultural purposes proposed by the European Union and by the EPA.     

Comparison of extraction and derivatization methods for fatty acid analysis in solid environmental matrixes

This study involved comparison of different extraction and derivatization methods for determining FAs in soil and in four highly organic matrixes (cattle manure, pig slurry, compost, and vermicompost), by application of a multifactor categorical design. Although some studies have been carried out regarding the application of FA analysis to highly organic matrixes, comparison and verification are still required to test which methods of extraction and derivatization of FAs function best for these matrixes. We compared three extraction methods (one in which the same extraction mixture as used in the Folch method was employed, a modification of the Bligh and Dyer method, and a microwave-assisted extraction) and two derivatization procedures (alkaline methanolysis and derivatization with trimethylsulfonium hydroxide (TMSH)). The highest yields of FAs belonging to different structural classes, and of individual FAs used as microbial biomarkers were obtained by application of the same extraction mixture as in the Folch method and use of TMSH as derivatization agent. These methods also involved a significant reduction in the complexity and time involved in sample preparation.     

Alkoxy-derived multiscale porous TiO<Subscript>2</Subscript> gels probed by ultra-small-angle X-ray scattering and small-angle X-ray scattering

Titania (TiO₂) monoliths with well-defined bicontinuous macropores and gel skeletons were prepared through the alkoxy-derived sol–gel process accompanied by spinodal decomposition, and the structural evolution during evaporation drying and heat treatment was probed by a combination of ultra-small-angle X-ray scattering and small-angle X-ray scattering. X-ray scattering profiles of wet and dried gels revealed that microporous structures related to the existence of primary particles are present in the gel skeletons at the wet stage and are preserved during drying. Additionally, it is found that the primary particles swollen in the wet condition are dried to compact aggregates to produce the smooth surface of gel skeletons. Upon heating at 400 °C, the particle–particle correlation associated with regularity of mesostructures is enhanced. From nitrogen adsorption–desorption measurements, the average pore size is less than 1 nm in the dried gel and increases to 3.1 nm by the heat treatment. Homogeneous growth of primary particles due to interparticle-polycondensation reaction is responsible for the increased size and uniform distribution of mesopores in the heat-treated gel.