Functional biodiversity and plasticity of methanogenic biomass from a full-scale mesophilic anaerobic digester treating nitrogen-rich agricultural wastes

The effect of ammonia on methanogenic biomass from a full-scale agricultural digester treating nitrogen-rich materials was characterized in batch activity assays subjected to increasing concentrations of total ammonia N. Acetotrophic and methanogenic profiles displayed prolonged lag phases and reduced specific activity rates at 6.0 gN-TAN L−1, though identical methane yields were ultimately reached. These results agreed with the expression levels of selected genes from bacteria and methanogenic archaea (qPCR of 16S rRNA and mrcA cDNA transcripts). Compound-specific isotope analysis of biogas indicated that ammonia exposure was associated to a transition in methanogenic activity from acetotrophy at 1.0 gN-TAN L−1 to intermediate and complete hydrogenotrophy at 3.5 and 6.0 gN-TAN L−1. Such pattern matched the results of 16S-Illumina sequencing of genes and transcripts in that predominant methanogens shifted, along with increasing ammonia, from the obligate acetotroph Methanosaeta to the hydrogenotrophic Methanoculleus and the poorly understood methylotrophic Methanomassiliicoccus. The underlying bacterial community structure remained rather stable but, at 6.0 gN-TAN L−1, the expression level increased considerably for a number of ribotypes that are related to potentially syntrophic genera (e.g. Clostridium, Bellilinea, Longilinea, and Bacteroides). The predominance of hydrogenotrophy at high ammonia levels clearly points to the occurrence of the syntrophic acetate oxidation (SAO), but known SAO bacteria were only found in very low numbers. The potential role of the identified bacterial and archaeal taxa with a view on SAO and on stability of the anaerobic digestion process under ammonia stress has been discussed.info:eu-repo/semantics/acceptedVersio

Acidification and solar drying of manure-based digestate to produce improved fertilizing products

The increase in energy and fertilizer consumption makes it necessary to develop sustainable alternatives for agriculture. Anaerobic digestion and digestates appeared to be suitable options. However, untreated digestates still have high water content and can increase greenhouse gas emissions during storage and land application. In this study, manure-derived digestate and solid fraction of digestate after separation were treated with a novel solar drying technology to reduce their water content, combined with acidification to reduce the gaseous emissions. The acidified digestate and acidified solid fraction of digestate recovered more nitrogen and ammonia nitrogen than their respective non-acidified products (1.5–1.3 times for TN; 14 times for TAN). Ammonia and methane emissions were reduced up to 94% and 72% respectively, compared to the non-acidified ones, while N2O increased more than 3 times. Dried digestate and dried acidified digestate can be labeled as NPK organic fertilizer regarding the European regulation, and the dried solid fraction and the improved dried acidified solid fraction can be labeled as N or P organic fertilizer. Moreover, plant tests showed that N concentrations in fresh lettuce leaves were within the EU limit with all products in all the cases. However, zinc concentration appeared to be a limitation in some of the products as their concentration exceeded the European legal limits.This work was funded by the European Union under the Circular Agronomics project (H2020 research and innovation project Nº.773649) and Nutry2Cycle project (H2020 research and innovation project Nº.773682). IRTA thanks the support of the CERCA Program and the Consolidated Research Group TERRA (ref.2017SGR1292), both from the Generalitat de Catalunya. L. Morey thanks the financial support of AGAUR, of the Generalitat de Catalunya (grant reference number 2019FI_B00694). We would like to thank the help of Celia Segura Godoy and Pau Berenguer i Planas during the sampling campaigns.Peer ReviewedPostprint (published version

Aberrant DNA Methylation of Matrix Remodeling and Cell Adhesion Related Genes in Pterygium

10.1371/journal.pone.0014687PLoS ONE62

Molecular Effects of Doxycycline Treatment on Pterygium as Revealed by Massive Transcriptome Sequencing

Pterygium is a lesion of the eye surface which involves cell proliferation, migration, angiogenesis, fibrosis, and extracellular matrix remodelling. Surgery is the only approved method to treat this disorder, but high recurrence rates are common. Recently, it has been shown in a mouse model that treatment with doxycycline resulted in reduction of the pterygium lesions. Here we study the mechanism(s) of action by which doxycycline achieves these results, using massive sequencing techniques. Surgically removed pterygia from 10 consecutive patients were set in short term culture and exposed to 0 (control), 50, 200, and 500 µg/ml doxycycline for 24 h, their mRNA was purified, reverse transcribed and sequenced through Illumina’s massive sequencing protocols. Acquired data were subjected to quantile normalization and analyzed using cytoscape plugin software to explore the pathways involved. False discovery rate (FDR) methods were used to identify 332 genes which modified their expression in a dose-dependent manner upon exposure to doxycycline. The more represented cellular pathways included all mitochondrial genes, the endoplasmic reticulum stress response, integrins and extracellular matrix components, and growth factors. A high correlation was obtained when comparing ultrasequencing data with qRT-PCR and ELISA results

Upgrading the temperature-phased anaerobic digestion of waste activated sludge by ultrasonic pretreatment

10 páginas; 5 figuras; 5 tablasBiochemical methane potential assays were performed to assess the influence of ultrasonic pretreatment on the temperature-phased anaerobic digestion (TPAD) of waste-activated sludge (WAS). Ultrasound (specific energy = 3380 kJ kg−1 TS) was applied to the WAS before the thermophilic stage or to the effluent after the thermophilic stage. In addition, a control system without pretreatment was also carried out. No significant differences were found in the overall performance of the TPAD process, but different behaviours were observed between the thermophilic and mesophilic stages. Total methane production was enhanced by more than 50% and the volatile solid removal increased by 13% in comparison to the TPAD control process. Finally, a previously defined kinetic model was applied successfully to the experimental data and showed an excellent fit.The authors wish to express their gratitude to the Spanish Ministry for the Environment, Rural Affairs and Marine Policy (Project 148/PC08/3-04.3) for providing financial support and to the 2008–2011 Research Plan of the University of Cadiz for supporting the first author through the ‘Contrato Puente para Doctores’.Peer reviewe

Closing nutrient loops in a maize rotation. Catch crops to reduce nutrient leaching and increase biogas production by anaerobic co-digestion with dairy manure

Three catch crop species, ryegrass, forage rape and black oat, were grown between successive rotations of maize to reduce nitrogen leaching due to maize fertilization with digested dairy manure. Catch crops showed a high nutrient uptake, but with a wide range, depending on the year and the specie. Ensiling was shown to be a feasible storing method increasing catch crop methane production per hectare between 14–36% compared with fresh catch crop. In semi-continuous co-digestion experiments, methane production was increased between 35–48%, in comparison with anaerobic digestion of dairy manure alone. Catch crops were shown to be a good co-substrate, being a sustainable option to prevent leaching of nutrients to the environment, thus closing the loops from production to utilization by optimal recycling measures.info:eu-repo/semantics/publishedVersio

Hygienization performances of innovative sludge treatment solutions to assure safe land spreading

n/