Development of functional consortia for the pretreatment of compostable lignocellulosic waste: A simple and effective solution to a large-scale problem.

Microorganisms drive the degradation of organic matter thanks to their enzymatic versatility. However, the structure of lignocellulose poses a great challenge for the microbiota inhabiting a compost pile. Our purpose was to increase the biodegradability of vegetable waste in the early stages of the composting process by applying a microbial consortium with lignocelllulolytic capacity. For this, a previous screening was performed among the culturable microbiota from different composting processes to find inoculants with ligninocellulolytic activity. Selected strains were applied as a pure culture and as a microbial consortium. The starting material was composed of tomato plant and pruning remains mixed in a ratio (50:50 v/v), whose humidity was adjusted to around 65%. To determine the ability of both treatments to activate the biodegradation of the mixtures, moisture, organic matter, ash, C/N ratio, 4-day cumulative respirometric index (AT4) and degradation rates of cellulose, hemicellulose and lignin were evaluated. Subsequently, a real composting process was developed in which the performance of the microbial consortium was compared with the composting process without inoculum (control). According to our tests, three microbial strains (Bacillus safensis, Bacillus licheniformis and Fusarium oxysporum) were selected. The results showed that the application of the bacteria strains at low doses (104 CFU g-1 on the complete residual material of the pile) resulted in higher rates of lignocelullose degradation after 10 days of treatment compared to that observed after application of the fungus in pure culture or untreated controls. The implementation of the strategy described in this work resulted in obtaining compost with better agronomic quality than the uninoculated controls. Therefore, the application of this consortium could be considered as an interesting tool for bioactivation of lignocellulosic waste prior to the composting process.

Sustainable approach to the control of airborne phytopathogenic fungi by application of compost extracts.

The search for new sustainable alternatives for plant disease control has gained interest in the last decades. Compost extracts are nowadays considered a remarkable alternative to agrochemicals due to their biopesticidal properties. However, these properties could be affected by the different variables of extraction protocols and by starting compost. This work focused on the physicochemical and biological characterization of compost extracts obtained from a wide range of composted materials and different extraction protocols (CEP). CEP-1 and CEP-4 involved incubation at 20 °C for 48 h and 14 days, respectively; CEP-2 incubation for 24 h at 40 °C; while CEP-3 were incubated for 12 h at 70 °C. Electrical conductivity, pH, total organic carbon (TOC) and phenolic content were determined as well as the actinobacterial count and enzyme profiles related to plant pathogen suppression. Additionally, the influence of the different materials and protocols on the in vitro growth inhibition of Alternaria alternata and Botrytis cinerea was determined. The starting materials and extraction protocols significantly influenced the physicochemical and biological characteristics of extracts. Treatments based on long incubation times at 20 °C, as well as those based on short incubation times at 40 °C, resulted in extracts with increased suppressive properties. However, extracts derived from CEP-3 protocol were characterized by high phenolic and TOC content, low functional biodiversity, and a more discreet antagonistic capacity. Therefore, the development and optimization of suitable extraction protocols could lead to compost extracts with increased phytoprotective capacities, thus becoming an effective and sustainable alternative to chemical inputs.

Mitigation of phytotoxic effect of compost by application of optimized aqueous extraction protocols.

The abuse of chemical fertilizers in recent decades has led the promotion of less harmful alternatives, such as compost or aqueous extracts obtained from it. Therefore, it is essential to develop liquid biofertilizers, which in addition of being stable and useful for fertigation and foliar application in intensive agriculture had a remarkable phytostimulant extracts. For this purpose, a collection of aqueous extracts was obtained by applying four different Compost Extraction Protocols (CEP1, CEP2, CEP3, CEP4) in terms of incubation time, temperature and agitation of compost samples from agri-food waste, olive mill waste, sewage sludge and vegetable waste. Subsequently, a physicochemical characterization of the obtained set was performed in which pH, electrical conductivity and Total Organic Carbon (TOC) were measured. In addition, a biological characterization was also carried out by calculating the Germination Index (GI) and determining the Biological Oxygen Demand (BOD5). Furthermore, functional diversity was studied using the Biolog EcoPlates technique. The results obtained confirmed the great heterogeneity of the selected raw materials. However, it was observed that the less aggressive treatments in terms of temperature and incubation time, such as CEP1 (48 h, room temperature (RT)) or CEP4 (14 days, RT), provided aqueous compost extracts with better phytostimulant characteristics than the starting composts. It was even possible to find a compost extraction protocol that maximize the beneficial effects of compost. This was the case of CEP1, which improved the GI and reduced the phytotoxicity in most of the raw materials analyzed. Therefore, the use of this type of liquid organic amendment could mitigate the phytotoxic effect of several composts being a good alternative to the use of chemical fertilizers.

Revisiting the succession of microbial populations throughout composting: A matter of thermotolerance.

Composting has been traditionally considered a process in which a succession of mesophilic and thermophilic microbial populations occurs due to temperature changes. In order to deepen in this model, 1380 bacterial and fungal strains (the entire culturable microbiota isolated from a composting process) were investigated for their ability to grow across a wide range of temperatures (20 to 60 °C). First, qualitative tests were performed to establish a thermal profile for each strain. Then, quantitative tests allowed ascertaining the extent of growth for each strain at each of the tested temperatures. The identity of the isolates enabled to position them taxonomically and permitted tracking the strains throughout the process. Results showed that 90% of the isolates were classified as thermotolerant (they grew at all tested temperatures). Only 9% and 1% of the studied strains showed to be strictly mesophilic or thermophilic, respectively. Firmicutes exhibited the greatest thermal plasticity, followed by Actinobacteria and Ascomycota. Most of the Proteobacteria and all Basidiomycota strains were also able to grow at all the assayed temperatures. Thermotolerance was clearly demonstrated among the composting microbiota, suggesting that the idea of the succession of mesophilic and thermophilic populations throughout the process might need a reassessment.

Uncovering new indicators to predict stability, maturity and biodiversity of compost on an industrial scale.

Currently, the metagenomic study of the composting process has gained great importance since it has allowed the identification of the existence of microorganisms that, until now, had not been isolated during the process by traditional techniques. However, it is still complex to determine which bioindicators could reveal the degree of maturity and stability of a particular compost. Thereby, the main objective of this work was to demonstrate the possible correlation between traditional parameters of maturity and stability of compost, with other indicators of biodiversity in products highly heterogeneous from composting processes on an industrial scale. The results demonstrated the enormous influence of the raw materials in characterizing the products obtained. Even so, important relationships were established between the Chao1 and Shannon indexes, and certain parameters related to the maturity, stability and toxicity of the samples, such as nitrification index, humification rate, phenolic content, germination index or oxygen consumption.

Evaluating the influence of raw materials on the behavior of nitrogen fractions in composting processes on an industrial scale.

Success of composting as an ecological technology for organic waste management has allowed its implementation in the current circular economy models. However, composting on an industrial scale often shows drawbacks and peculiarities. In this work, a comparative analysis of 15 industrial composting facilities was carried out in which different anthropogenic organic waste were processed. Results showed that composting process on an industrial scale did not always evolve in a standard way. Monitoring parameters as well as enzymatic activity depended largely on the raw materials and were strongly linked to the transformation of nitrogen fractions. Despite the heterogeneity of the processes and raw materials, microbial activity managed to the optimal biotransformation, obtaining products that comply with the agronomic quality standards. This work represents a breakthrough in composting and provides new knowledge for better management of this process on an industrial scale.

Enzymatic profiles associated with the evolution of the lignocellulosic fraction during industrial-scale composting of anthropogenic waste: Comparative analysis.

In the new European Waste Law, composting is proposed as one of the best options to properly manage organic waste of anthropogenic origin. Currently, the massive generation of this type of waste, as well as its heterogeneity, makes difficult in many cases control this process of degradation on an industrial scale. In this work, 15 facilities were selected based on 5 types of organic waste: Urban Solid Waste, Vegetable Waste, Sewage Sludges, Agrifood Waste and "Alpeorujo". The samples were collected in different thermal phases. The results revealed very different physicochemical and enzymatic profiles, as well as different degrees of humification depending on the process and the raw materials. However, parameters such as ß-glucosidase, amylase, lignin/holocellulose ratio and humification rate showed similar trends in all cases. All of them could act as important indicators to evaluate the quality of a composting process, despite the heterogeneity of the starting materials.