Evolution of enzymatic activities and carbon fractions throughout composting of plant waste.

Many alternatives for the proper disposal of horticultural plant wastes have been studied, and composting is one of the most attractive due to its insignificant environmental impact and low cost. The quality of compost for agronomical use is related to the degree of organic matter maturation and stabilization. Traditional parameters as well as temperature, ratio C/N, cationic exchange capacity, extractable carbon, or evolution of humificated substances have been successfully used to assess compost maturity and stability. However, microorganisms frequently isolated during composting release a wide range of hydrolytic enzymes, whose activity could apparently give interesting information on the rate of decomposition of organic matter and, therefore, on the product stability. The aim of this work was to study the evolution of some important enzymatic activities during composting of agricultural wastes and their comparison with other chemical parameters commonly employed as quality and maturity indexes, to establish a relationship between the degradation intensity of specific organic carbon fractions throughout the process. In this work, the chemical and biochemical parameters of plant wastes were studied along a composting process of 189 days to evaluate their importance as tools for compost characterization. Results showed an intense enzymatic activity during the first 2-3 weeks of composting (bio-oxidative phase), because of the availability of easily decomposable organic compounds. From a biological point of view, a less intense phase was observed between second and third month of composting (mesophilic or cooling phase). Finally, chemical humification parameters were more closely associated with the period between 119 and 189 days (maturation phase). Significant correlations between the enzymatic activities as well as between enzyme activities and other more traditional parameters were also highlighted, indicating that both kind of indexes can be a reliable tool to determine the degree of stability and maturation of horticultural plant wastes based-compost.

Microbial-induced MnO2 precipitation in a carbonate coastal aquifer.

A study was carried out to identify biogeochemical reactions along a transect of a coastal dolomitic aquifer. In this transect, the physicochemical parameters of the groundwater as well as the microbial composition of samples taken at different depths and salinities were measured. Many of the dissolved ions measured in the groundwater follow a pattern that reflects the distribution of the water masses (fresh, interface and salt) in the aquifer, while others such as Ca and Mg ions deviate from this trend by identifying the zones of maximum dissolution of the carbonate matrix. The concentrations of minor ions, such as Fe and Mn, also follow a singular pattern, with maximum concentrations in the reducing zones of the aquifer and lower values in the oxidizing zones. Precipitates of Mn oxides along with other metals, such as Fe, Ba, Zn and Ni, were observed in the saline zone displaying oxidizing conditions close to the coastline, where a continuous core was recovered. This zone, which is located below the freshwater-seawater mixing zone and features percentages of seawater higher than 80 %, is characterized by the presence of Marinobacter as the predominant genus. These bacteria are also related to the formation of Mn-rich polymetallic oxides in other contexts such as the ocean floor (Wang et al., 2012; Cao et al., 2021). All in all, a biogeochemical reaction model is proposed that describes the formation of these oxides in areas close to the discharge zone of coastal aquifers. To do this, it has been necessary to integrate the results obtained from geochemical, hydrogeological and microbiological information.

Evolution of the pathogen content during co-composting of winery and distillery wastes.

The aim of this study was to monitor some microbial indicators and pathogen contents (sulphite reducers clostridia, total enterobacteriaceae, total coliforms, faecal coliforms (Escherichia coli), enterococci, Staphylococcus aureus and Salmonella spp.) throughout the co-composting of wastes from the winery and distillery industry with other organic residues, as well as the effect of the composting system used. Seven different piles using mixtures of winery-distillery wastes with other organic materials were prepared. P1 and P2 were made using grape stalk (GS), grape marc (GM), exhausted grape marc (EGM) and sewage sludge (SS), whereas in P3 and P4 were also used exhausted grape marc with cow manure (CW) and poultry manure (PM), respectively, using the Rutgers system. Additionally, P2 was watered with vinasse (V). The rest of piles (P5, P6 and P7) were prepared with grape marc, exhausted grape marc, cow manure and poultry manure, using the turning system. The effectiveness of the composting process to reduce the pathogen content was higher in the static aerated piles than in those elaborated with the turning. The relatively high temperatures (50-60 degrees C) reached in some of the piles produced a notable decrease in some microbial groups, such as total and faecal coliforms (E. coli), but the characteristics of the raw materials used notably influenced the pathogen contents of the end-product.

Effect of horticultural waste composting on infected plant residues with pathogenic bacteria and fungi: integrated and localized sanitation.

The aim of this work was to study the effect of composting on the viability of plant pathogenic fungi and bacteria. The research consisted of pilot-scale composting of horticultural waste in compost windrows. Studies were carried out on vegetable residues infected with plant pathogenic microorganisms included by either integrated or localized infection. In the first case, the plant pathogen viability was investigated when infected material was mixed throughout compost, while the localized infection was used to study the effect of the composting process on plant waste spot-inoculated with pathogenic microorganisms. Results for localized sanitation showed the total elimination of all tested phytopathogens between 48 and 120 h after composting began. In this case significant differences were observed in relation to 9 different zones in the pile. The disappearance of these microorganisms was similar when all plant waste included in the windrow was infected (integrated infection). Additionally, the results obtained confirmed that the bacteria showed a greater capacity to persist during composting than the fungi. Composting is therefore considered a useful method for recycling horticultural waste and eliminating phytopathogenic bacteria and fungi that inhabit this kind of residue.

Effect of inoculation in composting processes: modifications in lignocellulosic fraction.

Three microbial isolates, identified as Bacillus shackletonni, Streptomyces thermovulgaris and Ureibacillus thermosphaericus were tested as inoculants in composting processes in relation to their capacity to improve lignocellulose degradation. Different wastes from agricultural activities were used as raw material for the heaps: pepper plant waste (PPW) as the main component and olive-oil mill waste (OMW), almond shell (AS), pruning waste (PW) and rice straw (RS) as additives. Cellulose was more extensively degraded than hemicellulose and lignin, although the use of inoculants (B. shackletonni and S. thermovulgaris) improved the action of the autochthonous microbiota just in the AS heaps. A higher efficiency was observed for lignin, since lower concentrations of this polymer were detected in the inoculated heaps in relation to control heaps. U. thermosphaericus was the most efficient microorganism since inoculation with this strain decreased the final lignin content in a range between 17.23% and 24.34%. S. thermovulgaris and B. shackletonni led to a higher reduction of the lignin levels in the OMW and PW heaps (14.25% and 19.07% less lignin than control heaps) and OMW (13%), respectively. The composting process can therefore be improved by means of inoculation if the microorganisms used for this purpose are appropriate for the characteristics of the raw material.

Laboratory study of inocula production for composting processes.

Six ligno-cellulolytic fungi were tested regarding to examine their capability to grow on agricultural wastes and produce inocula for composting. Two residues were used: pepper plant wastes and almond shell residues. Results showed the latter as the most adequate substrate for growth of fungi tested. On the contrary, Trichoderma koningii, as well as HLC1 and HLC3, both fungi isolated from almond shell wastes, were able to persist in pepper plant wastes. Modifications of aeration and pH significantly influenced growth of Coriolus versicolor, HLC1 and Phanerochaete. flavido alba and P. flavido alba and Phlebia radiata, respectively, while P. flavido alba was the only microorganism whose growth was not significantly altered by temperature. In competitive assays, where fungi were growth together with other species, growth of both microorganisms isolated from almond shell residues, HLC1 and HLC3, were stimulated while T. koningii showed the better results in sterile conditions.

Dynamics of bacterial microbiota during lignocellulosic waste composting: Studies upon its structure, functionality and biodiversity.

An intensive isolation program carried out in three replicated composting piles allowed the identification of the resident and transient components of the composting microbiome. More than 4000 bacterial strains were isolated, enzymatically characterized and identified by partial sequencing of their 16S rRNA gene. While microorganisms isolated under mesophilic conditions were prominent throughout the process, thermophilic stages gathered the highest total counts and spore-forming bacteria prevailed at the bio-oxidative phase of composting. Enzymatic capabilities related to the degradation of polymeric materials were exhibited by most of the isolates and as a result of these activities, more soluble compounds could be made available to the entire composting microbiota. A high proportion of isolates showed to be thermotolerant as they were detected at mesophilic and thermophilic phases. Isolated strains belonged to 187 bacterial species. Biodiversity was greater at the central stages of composting and mesophilic, thermophilic and cooling phases shared 50% of species.

Enhanced turnover of organic matter fractions by microbial stimulation during lignocellulosic waste composting.

Enhanced organic matter turnover was detected in lignocellulosic composting piles inoculated with microorganisms specifically capable of decomposing polymeric compounds. In comparison to uninoculated piles, the following results were obtained in the inoculated piles: degradation of hemicellulose, cellulose and lignin were 28%, 21% and 25% respectively higher. Total organic matter, total sugars and phenolic compounds also decreased more intensely. Greater amounts of soluble organic carbon, reducing sugars and soluble proteins were available to the composting microbiota. Recycling of organic to inorganic nitrogen was improved and humification was more intense and earlier attained. Microbial community structure was also affected by inoculation. It was initially thought that these effects were due to enzymatic capabilities of inoculants, however, microbial counts, especially those corresponding to functional groups, revealed that inoculation induced a true stimulation of microbial growth and activity in the entire composting microbiota which was actually responsible for all the beneficial effects reported here.

The effect of aeration on the biotransformation of lignocellulosic wastes by white-rot fungi.

The mineralisation and the humification of organic matter (OM) in sterile horticultural plant wastes inoculated with Coriolus versicolor or Phanerochaete flavido-alba was investigated under different aeration rates in order to determine their efficacy as potential inoculants for composting. The change in elemental composition, lignin content and OM fractions was analysed during a 90-day incubation. Both fungi degraded 30% of lignin at low aeration rates. Different aeration rates led to significant changes in OM mineralisation induced by C. versicolor, but did not have noticeable effect on P. flavido-alba activity. The mineralisation was more effectively carried out by P. flavido-alba than by C. versicolor. Lignin degradation and the linked humification process were equally achieved by the two fungi and were enhanced in aerated conditions. The fungi analysed may facilitate the composting of lignocellulosic wastes by means of an increase in substrate bioavailability and OM humification.

Microbial population dynamics and enzyme activities in composting processes with different starting materials.

A biological comparison based on differences in the starting material for composting processes was made. Mesophilic aerobic microbiota, fungi, actinomycetes and hemicellulolytic microorganisms reached significantly higher levels in the MSW final product. The population of cellulolytic microorganisms did not show a clear trend, although it was more numerous in the HW piles. Counts for N(2)-fixing and ammonifying bacteria were significantly higher in the SS pile at the early stages of the process, while populations tended to become equal as time progressed. The lowest populations were detected for nitrifying bacteria, with higher but not always significant levels for the SS pile. beta-Glucosidase and phosphatase activities were higher in the SS pile at the early stages. Protease reached its maximum activity during the bio-oxidative phase and final stages in the HW and MSW piles, respectively. Dehydrogenase activity, with an occasional high level for the MSW at the beginning of the process, was almost inexistent since the end of the bio-oxidative phase. On the contrary, urease showed higher levels at the final stage of the process, with the MSW pile showing the greatest levels most of the time. According to these results, the nature of the starting material causes differences in biological parameters.

Properties of polysaccharide produced by Azotobacter vinelandii cultured on 4-hydroxybenzoic acid.

AIMS: Characterization of the exopolysaccharide produced by Azotobacter vinelandii grown on 4-hydroxybenzoic acid (EPS I), and the comparison between this exopolysaccharide and commercial alginate, constituted the main objective of this work. METHODS AND RESULTS: Total carbohydrates, uronic acids, acetyl and pyruvyl groups and proteins were determined by colorimetric methods and composition was confirmed by Nuclear Magnetic Resonance studies. Rheological properties were analysed under different physical and chemical conditions. Results showed differences between EPS I and commercial alginate, in relation to both composition and viscosity. Higher amount of guluronnosyl residues were found in EPS I, whereas commercial alginate contained the same proportion of mannuronosyl and guluronnosyl residues. In accordance with this result, EPS I gave rise to solutions of higher viscosity than commercial alginate, although solutions of this polysaccharide showed greater stability when conditions were altered. CONCLUSIONS: The exopolysaccharide produced by A. vinelandii grown on 4-hydroxybenzoic acid showed a different composition in comparison with commercial alginate, which leads to higher viscosity values for the aqueous solutions of EPS I. SIGNIFICANCE AND IMPACT OF STUDY: This work describes for the first time the characteristics of an exopolysaccharide produced by A. vinelandii from 4-hydroxybenzoic acid, a substrate rarely used as sole carbon source.

Physiology of exopolysaccharide production by Azotobacter vinelandii from 4-hydroxybenzoic acid.

The relationship between exopolysaccharide (EPS) production by Azotobacter vinelandii ATCC 12837 from 4-hydroxybenzoic acid as sole carbon source and other physiological parameters was investigated. In relation to growth, Azotobacter needed more time in 4-hydroxybenzoic acid to reach levels of biomass similar to those obtained when sugars were used, although the phenolic compound led to a more extensive exponential phase. The encystment process was initiated after cells had grown for 24 h, in which small amounts of EPS were synthesized and poly-beta-hydroxybutyrate (PHB) accumulation began. Both polymers, EPS and PHB, showed a similar evolution with time, as well as the formation of cysts, which points out the existence of a relation between these parameters. This was corroborated by a statistical study, in which significant correlations (P<0.05) were observed when each parameter was compared to the two others.

Influence of nutritional and environmental factors on polysaccharide production by Azotobacter vinelandii cultured on 4-hydroxybenzoic acid.

The capacity of 4-hydroxybenzoic acid to support exopolysaccharide (EPS) biosynthesis was investigated. Carbon source concentration, nitrogen supplementation, and other nutritional and environmental factors were optimized to obtain maximal EPS recovery. Higher EPS yields were obtained in nitrogen-free media amended with 20-30 mM 4-hydroxybenzoic acid. In general, modifications in inorganic salt concentration did not alter EPS production, except in the case of magnesium ions. Increased levels of this cation were correlated to greater EPS yields. Production was strongly influenced by certain environmental factors. Optimal values of 34 degrees C, 80 rpm and neutral or slightly basic conditions were selected. Under these conditions, more than 25% of the carbon source supplied was converted to EPS and the production was improved about 42% in comparison to that observed in the initial media.

Temperature effect on Fusarium oxysporum f.sp. melonis survival during horticultural waste composting.

AIMS: The aim of this work was to study the effect of high temperatures generated during composting process, on the phytopathogen fungus Fusarium oxysporum f.sp. melonis. This investigation was achieved by both in vivo (semipilot-scale composting of horticultural wastes) and in vitro (lab-scale thermal treatments) assays. METHODS AND RESULTS: Vegetable residues infected with F. oxysporum f.sp. melonis were included in compost piles. Studies were conducted in several compost windrows subjected to different treatments. Results showed an effective suppression of persistence and infective capacity, as this process caused complete fungal elimination after 2-3 days of composting. In order to confirm the effect of high temperature during this process, in vitro experiments were carried out. Temperature values of 45, 55 and 65 degrees C were tested. All three treatments caused the elimination of fungal persistence. Treatment at 65 degrees C was especially effective, whereas 45 degrees C eliminated fungal persistence only after 10 days. CONCLUSIONS: The composting process is an excellent alternative for the management of plant wastes after harvesting, as this procedure is able to suppress infective capacity of several harmful phytopathogens such as F. oxysporum f.sp. melonis. SIGNIFICANCE AND IMPACT OF THE STUDY: Fusarium oxysporum f.sp. melonis is a plant pathogen fungus specially important in the province of Almería (south-east Spain), where intensive greenhouse horticulture is very extended. High temperatures reached during composting of horticultural plant wastes ensure the elimination of phytopathogen microorganisms such as F. oxysporum f.sp. melonis from vegetable material, providing an adequate hygienic quality in composts obtained.