Unraveling the potential and constraints associated with corn steep liquor as a nutrient source for industrial fermentations.

Costly complex media components such as yeast extract and peptone are still widely used in industrial bioprocesses, despite their ill-defined composition. Side stream products such as corn steep liquor (CSL) present a compelling economical alternative that contains valuable nutrients required for microbial growth, that is, nitrogen and amino acids, but also vitamins, trace elements, and other minerals. However, as a side stream product, CSL may be subject to batch-to-batch variations and compositional heterogeneity. In this study, the Respiration Activity MOnitoring System designed for shake flasks (RAMOS) and 96-well microtiter plates (µTOM) were applied to investigate the potential and constraints of CSL utilization for two model microorganisms: E. coli and B. subtilis. Considering the dry substance content of complex nutrients involved, CSL-based media are more efficient in biomass production than the common lysogeny broth (LB) medium, containing 5 g/L yeast extract, 10 g/L peptone, and 5 g/L NaCl. At a glucose to CSL (glucose/CSL, g/g) ratio of 1/1 (g/g) and 2/1 (g/g), a secondary substrate limitation occurred in E. coli and B. subtilis cultivations, respectively. The study sheds light on differences in the metabolic activity of the two applied model organisms between varying CSL batches, which relate to CSL origin and production process, as well as the effect of targeted nutrient supplementation. Through a targeted nutrient supplementation, the most limiting component of the CSL-glucose medium used for these applied model microorganisms was identified to be ammonium nitrogen. This study proves the suitability of CSL as an alternative nutrient source for E. coli and B. subtilis. The RAMOS and µTOM technique detected differences between CSL batches, allowing easy and early identification of varying batches. A consistent performance of the CSL batches in E. coli and B. subtilis cultivations was demonstrated.

Assembled mixed co-cultures for emerging pollutant removal using native microorganisms from sewage sludge.

The global pharmaceutical pollution caused by drug consumption (>100,000 tonnes) and its disposal into the environment is an issue which is currently being addressed by bioremediation techniques, using single or multiple microorganisms. Nevertheless, the low efficiency and the selection of non-compatible species interfere with the success of this methodology. This paper proposes a novel way of obtaining an effective multi-domain co-culture, with the capacity to degrade multi-pharmaceutical compounds simultaneously. To this end, seven microorganisms (fungi and bacteria) previously isolated from sewage sludge were investigated to enhance their degradation performance. All seven strains were factorially mixed and used to assemble different artificial co-cultures. Consequently, 127 artificial co-cultures were established and ranked, based on their fitness performance, by using the BSocial analysis web tool. The individual strains were categorized according to their social behaviour, whose net effect over the remaining strains was defined as 'Positive', 'Negative' or 'Neutral'. To evaluate the emerging-pollutant degradation rate, the best 10 co-cultures, and those which contained the social strains were then challenged with three different Pharmaceutical Active compounds (PhACs): diclofenac, carbamazepine and ketoprofen. The co-cultures with the fungi Penicillium oxalicum XD-3.1 and Penicillium rastrickii were able to degrade PhACs. However, the highest performance (>80% degradation) was obtained by the minimal active microbial consortia consisting of both Penicillium spp., Cladosporium cladosporoides and co-existing bacteria. These consortia transformed the PhACs to derivate molecules through hydroxylation and were released to the media, resulting in a low ecotoxicity effect. High-throughput screening of co-cultures provides a quick, reliable and efficient method to narrow down suitable degradation co-cultures for emerging PhAC contaminants while avoiding toxic metabolic derivatives.

Assessment of the antioxidative response and culturable micro-organisms of Lygeum spartum Loefl. ex L. for prospective phytoremediation applications.

Abundant plant species in arid industrial areas are mining phyto-resources for sustainable phyto-management. However, the association with their rhizosphere is still poorly known for phytoremediation purposes. This study aims to assess the heavy metals (HMs) and metalloids uptake of Lygeum spartum Loefl. ex L. growing in cement plant vicinity and screen associated culturome for potential phytoremediation use. Bioaccumulation factor (BAF), the translocation factor (TF), and the mobility ratio (MR) were studied along with four sites. Lipid peroxidation (MDA), free proline (Pro), Non-protein thiols (NPTs), and reduced glutathione (GSH) were tested for evaluating the plant antioxidative response. Bacteria and fungi associated with L. spartum Loefl. ex L. were identified by 16S rRNA and fungal internal transcribed spacer (ITS1-ITS2) gene sequencing. Our results showed an efficient uptake of As, Pb, and Zn and enhanced GSH (0.34 ± 0.03) and NPTs (528.7 ± 14.4 nmol g-1 FW) concentrations in the highly polluted site. No significant variation of Arbuscular Mycorrhizal Fungi (AMF) was found. Among 29 bacterial isolates, potential bioremediation were Bacillus simplex and Bacillus atrophaeus. Thus, L. spartum Loefl. ex L. and its associated microbiota have the potential for phytoremediation applications. Novelty statement: This work has been set in line with the investigation of the integrative biology of Lygeum spartum Loefl ex L. and the screening of its associated microbiome for potential phytoremediation applications. This work is the first work conducted in a cement plant vicinity investigating the associated fungi and bacteria of L. spartum Loefl. ex L. and been part of a sectorial research project since 2011, for assessing the impact of industrial pollution and recognizing the accumulation potential of plant species for further phyto-management applications.

High-Throughput Microbial Community Analyses to Establish a Natural Fungal and Bacterial Consortium from Sewage Sludge Enriched with Three Pharmaceutical Compounds.

Emerging and unregulated contaminants end up in soils via stabilized/composted sewage sludges, paired with possible risks associated with the development of microbial resistance to antimicrobial agents or an imbalance in the microbial communities. An enrichment experiment was performed, fortifying the sewage sludge with carbamazepine, ketoprofen and diclofenac as model compounds, with the aim to obtain strains with the capability to transform these pollutants. Culturable microorganisms were obtained at the end of the experiment. Among fungi, Cladosporium cladosporioides, Alternaria alternata and Penicillium raistrickii showed remarkable degradation rates. Population shifts in bacterial and fungal communities were also studied during the selective pressure using Illumina MiSeq. These analyses showed a predominance of Ascomycota (Dothideomycetes and Aspergillaceae) and Actinobacteria and Proteobacteria, suggesting the possibility of selecting native microorganisms to carry out bioremediation processes using tailored techniques.

Design of Bio-Absorbent Systems for the Removal of Hydrocarbons from Industrial Wastewater: Pilot-Plant Scale.

The objective of this study was the development and design of a treatment system at a pilot-plant scale for the remediation of hydrocarbons in industrial wastewater. The treatment consists of a combined approach of absorption and biodegradation to obtain treated water with sufficient quality to be reused in fire defense systems (FDSs). The plant consists of four vertical flow columns (bioreactors) made of stainless steel (ATEX Standard) with dimensions of 1.65 × 0.5 m and water volumes of 192.4 L. Each bioreactor includes a holder to contain the absorbent material (Pad Sentec polypropylene). The effectiveness of the treatment system has been studied in wastewater with high and low pollutant loads (concentrations higher than 60,000 mg L-1 of total petroleum hydrocarbons (TPH) and lower than 500 mg L-1 of TPHs, respectively). The pilot-plant design can function at two different flow rates, Q1 (180 L h-1) and Q2 (780 L h-1), with or without additional aeration. The results obtained for strongly polluted wastewaters showed that, at low flow rates, additional aeration enhanced hydrocarbon removal, while aeration was unnecessary at high flow rates. For wastewater with a low pollutant load, we selected a flow rate of 780 L h-1 without aeration. Different recirculation times were also tested along with the application of a post-treatment lasting 7 days inside the bioreactor without recirculation. The microbial diversity studies showed similar populations of bacteria and fungi in the inlet and outlet wastewater. Likewise, high similarity indices were observed between the adhered and suspended biomass within the bioreactors. The results showed that the setup and optimization of the reactor represent a step forward in the application of bioremediation processes at an industrial/large scale.

Evaluation of the Potential of Sewage Sludge Mycobiome to Degrade High Diclofenac and Bisphenol-A Concentrations.

One of the most challenging environmental threats of the last two decades is the effects of emerging pollutants (EPs) such as pharmaceutical compounds or industrial additives. Diclofenac and bisphenol A have regularly been found in wastewater treatment plants, and in soils and water bodies because of their extensive usage and their recalcitrant nature. Due to the fact of this adversity, fungal communities play an important role in being able to safely degrade EPs. In this work, we obtained a sewage sludge sample to study both the culturable and non-culturable microorganisms through DNA extraction and massive sequencing using Illumina MiSeq techniques, with the goal of finding degraders adapted to polluted environments. Afterward, degradation experiments on diclofenac and bisphenol A were performed with the best fungal degraders. The analysis of bacterial diversity showed that Dethiosulfovibrionaceae, Comamonadaceae, and Isosphaeraceae were the most abundant families. A predominance of Ascomycota fungi in the culturable and non-culturable population was also detected. Species such as Talaromyces gossypii, Syncephalastrum monosporum, Aspergillus tabacinus, and Talaromyces verruculosus had remarkable degradation rates, up to 80% of diclofenac and bisphenol A was fully degraded. These results highlight the importance of characterizing autochthonous microorganisms and the possibility of selecting native fungal microorganisms to develop tailored biotransformation technologies for EPs.

Assessment of bacterial and fungal communities in a full-scale thermophilic sewage sludge composting pile under a semipermeable cover.

Bacterial and fungal communities in a full-scale composting pile were investigated, with sewage sludge and a vegetal bulking agent as starting materials. Bacillales and Actinomycetales were predominant throughout the process, showing significant abundance. Ascomycota was the predominant fungal phylum during the thermophilic phase, with a shift to Basidiomycota at the end of the process. The bulking material was the principal contributor to both communities by the end of the process, with a signal above 50%. The presence of genera, such as Pedomicrobium, Ureibacillus and Tepidimicrobium at the end of the process, and Chaetomium and Arthrographis in the maturation phase, showed an inverse correlation with indicators of organic matter stabilisation. A semipermeable cover was an effective technology for excluding pathogens. These results indicate that changes in the microbial population and their interrelation with operational variables could represent a useful tool for monitoring composting processes.

Assessment of the diversity and abundance of the total and active fungal population and its correlation with humification during two-phase olive mill waste (''alperujo") composting.

Metagenomic and transcriptomic techniques applied to composting could increase our understanding of the overall microbial ecology and could help us to optimise operational conditions which are directly related with economic interest. In this study, the fungal diversity and abundance of two-phase olive mill waste ("alperujo") composting was studied using Illumina MiSeq sequencing and quantitative PCR, respectively. The results showed an increase of the fungal diversity during the process, with Ascomycota being the predominant phylum. Penicillium was the main genera identified at the mesophilic and maturation phases, with Debaryomyces and Sarocladium at the thermophilic phase, respectively. The fungal abundance was increased during composting, which confirms their important role during thermophilic and maturation phases. Some Basidiomycota showed an increased during the process, which showed a positive correlation with the humification parameters. According to that, the genus Cystofilobasidium could be used as a potential fungal biomarker to assess alperujo compost maturation.

Effect of Composting Under Semipermeable Film on the Sewage Sludge Virome.

The addition of compost from sewage sludge to soils represents a sustainable option from an environmental and economic point of view, which involves the valorisation of these wastes. However, before their use as a soil amendment, compost has to reach the quality levels according to the normative, including microbial parameters. Viruses are not included in this regulation and they can produce agricultural problems and human diseases if the compost is not well sanitised. In this study, we carried out the analysis of the viral populations during a composting process with sewage sludge at an industrial scale, using semipermeable cover technology. Viral community was characterised by the presence of plant viruses and bacteriophages of enteric bacteria. The phytopathogen viruses were the group with the highest relative abundance in the sewage sludge sample and at 70 days of the composting process. The diversity of bacterial viruses and their specificity, with respect to the more abundant bacterial taxa throughout the process, highlights the importance of the interrelations between viral and bacterial communities in the control of pathogenic communities. These results suggest the possibility of using them as a tool to predict the effectiveness of the process.

Leadership and Motivational Climate: The Relationship with Objectives, Commitment, and Satisfaction in Base Soccer Players.

The objective of the present study is to analyze non-professional soccer players' preferences regarding coach leadership style and motivational climate and to determine the relationship of these variables with players' satisfaction, sport commitment, and sport objectives. The participants were 151 players, aged between 10 and 24 years, divided into five categories: Alevín, Infantil, Cadet, Feminine, and Juvenile, all belonging to the Aragonese Soccer Federation. The participants completed questionnaires assessing their perception of their coach's leadership style, the team's motivational climate, their individual satisfaction, degree of sport commitment, and sport objectives. The results show that the leadership styles of training and instruction (M = 3.98, SD = 0.43) and positive feedback (M = 4.02, SD = 0.53) are the most valued by the players in all categories. The training and instruction leadership style had the highest correlations with task-oriented motivational climate (r = 0.40). The findings of the regression analysis show that a training and instruction leadership style and a task-oriented motivational climate significantly predict players' satisfaction (13.3%) and sport commitment (14.5%).

Effect of semi-permeable cover system on the bacterial diversity during sewage sludge composting.

Sewage sludge composting is a profitable process economically viable and environmentally friendly. In despite of there are several kind of composting types, the use of combined system of semipermeable cover film and aeration air-floor is widely developed at industrial scale. However, the knowledge of the linkages between microbial communities structure, enzyme activities and physico-chemical factors under these conditions it has been poorly explored. Thus, the aim of this study was to investigate the bacterial dynamic and community structure using next generation sequencing coupled to analyses of microbial enzymatic activity and culturable dependent techniques in a full-scale real composting plant. Sewage sludge composting process was conducted using a semi-permeable Gore-tex cover, in combination with an air-insufflation system. The highest values of enzymatic activities such as dehydrogenase, protease and arylsulphatase were detected in the first 5 days of composting; suggesting that during this period of time a greater degrading activity of organic matter took place. Culturable bacteria identified were in agreement with the bacteria found by massive sequencing technologies. The greatest bacterial diversity was detected between days 15 and 30, with Actinomycetales and Bacillales being the predominant orders at the beginning and end of the process. Bacillus was the most representative genus during all the process. A strong correlation between abiotic factors as total organic content and organic matter and enzymatic activities such as dehydrogenase, alkaline phosphatase, and ß-glucosidase activity was found. Bacterial diversity was strongly influenced by the stage of the process, community-structure change was concomitant with a temperature rise, rendering favorable conditions to stimulate microbial activity and facilitate the change in the microbial community linked to the degradation process. Moreover, results obtained confirmed that the use of semipermeable cover in the composting of sewage sludge allow a noticeable reduction in the process-time comparing to conventional open windrows.

Response of autochthonous microbiota of diesel polluted soils to land-farming treatments.

This study investigated the response of autochthonous microorganisms of diesel polluted soils to land-farming treatments. Inorganic NPK (nitrogen, phosphorous, and potassium) fertilizer and Ivey surfactant were applied alone or in combination as biostimulating agents. The study was carried out in experimental separated land-farming plots performed with two soils: a sandy clay soil with low biological activity and a sandy clay soil with higher biological activity, contaminated with two concentrations of diesel: 10,000 and 20,000mgkg(-1). Bacterial growth, dehydrogenase activity and CO2 production were the biological parameters evaluated. Non-metric multidimensional scaling analysis proved that moisture content showed a tendency related to microbial growth and that heterotrophic and degrading microorganisms had the best relationship. Initial biological activity of soil influenced the response with 11.1% of variability attributed to this parameter. Soils with low activity had higher degree of response to nutrient addition.

Bioremediation of diesel-polluted soil using biostimulation as post-treatment after oxidation with Fenton-like reagents: assays in a pilot plant.

The present study focuses on the remediation of diesel-polluted soil using modified Fenton treatment coupled with inorganic NPK fertilizer ("Fenton+NPK"). Studies were carried out in a pilot plant containing 1 m(3) of sandy soil contaminated with 20,000 mg kg(-1) of diesel, placed outdoors at a temperature ranging between 5 and 10 °C. Results showed that NPK-fertilizer as post-treatment stimulated culturable degrading bacteria and enhanced dehydrogenase activity. Fenton+NPK treatment increased total petroleum hydrocarbon (TPH) removal efficacy. Natural attenuation removed 49% of TPH in the surface layer, 23% of TPH in the non-saturated layer and 4% of the TPH in the saturated layer, while the percentage removed of TPH after Fenton+NPK treatment was 58%, 57% and 32% respectively. The results from our study showed that, immediately after soil contamination, occurred a specialization and differentiation of the bacterial community, but after this initial modification, no significant changes of bacterial diversity was observed under natural attenuation conditions. In contrast, when the Fenton's reagent was applied a reduction of the bacterial biodiversity was observed. However, the post-biostimulation did enhance the degrading microbiota and stimulated their degrading biological activity. In conclusion, biostimulation, as a post-treatment step in chemical oxidation, is an effective solution to remediate hydrocarbon-polluted sites.

Necesidades de investigación en carotenoides en América Latina / Investigation needs on carotenoids in Latin America

Muchos de los fitoquímicos bioactivos son muy importantes para mantener un buen estado de salud y de ellos los carotenoides han sido los más estudiados. Se conocen alrededor de 637 y se ha propuesto que posiblemente 70 de ellos puedan tener un papel en la salud humana. En cortes de cerebro humano se han encontrado 16 en relativamente alta cantidad. La mayoría de los estudios encuentran relación entre los carotenoides y varias enfermedades de tipo crónico no transmisibles, sobre todo con ciertos tipos de cáncer, enfermedades aterosclerósicas y enfermedades degenerativas del ojo. Esta relación esta mediada por los genes y la edad. Los carotenoides tienen interés científico y también económico para los países latinoamericanos, por lo que se propone hacer análisis en los productos tropicales, frutas y verduras más comunes, lo mismo que hacer estudios sobre su disponibilidad. Se sugiere un trabajo coordinado en el que se determinen los 7 carotenoides más relacionados con la salud en 40 ó 50 productos tropicales, lo mismo que comenzar a evaluar su disponibilidad metabólica. Se considera asimismo posible hacer estudios epidemiológicos basados en las poblaciones consumidoras de algunos alimentos ricos en carotenos para hacer análisis de riesgo comparativos. En algunos países de Latinoamérica, con el patrocinio de FAO e Infoods, y se han llevado a cabo algunas reuniones y cursos para iniciar este importante trabajo, lo mismo que para completar las tablas de composición de alimentos y hacer comparación entre países