Enzymatic Potential of Bacteria and Fungi Isolates from the Sewage Sludge Composting Process

The aim of this study was the isolation and characterisation of the fungi and bacteria during the composting process of sewage sludge under a semipermeable membrane system at full scale, in order to find isolates with enzymatic activities of biotechnological interest. A total of 40 fungi were isolated and enzymatically analysed. Fungal culture showed a predominance of members of Ascomycota and Basidiomycota division and some representatives of Mucoromycotina subdivision. Some noticeable fungi isolated during the mesophilic and thermophilic phase were Aspergillus, Circinella, and Talaromyces. During the maturation phase, some lignin modifying enzyme producers, like Purpureocillium, Thielavia, Bjerkandera, or Dichotomyces, were found. Within this group, Thielavia and Bjerkandera showed high activity with production of laccases and peroxidases. In the bacterial culturome, a total of 128 strains were selected and enzymatically analysed. Bacillales, Actinomycetales, Pseudomonadales, and Lactobacillales were the orders most represented in culture-bacteria. Bacillus pumilus, B. stratosphericus, B. safensis, and Pseudomonas formosensis were the species most e cient in enzyme production, particularly peroxidases, polyphenol oxidases ammonifying activity, and amylases. These results showed that sewage sludge composting piles could represent a source of microorganisms which have adapted to adverse conditions.Junta de Andalucía RNM-7370 CTM2017-84332-RJunta de Andalucía RYC-2013-1248

First Report of in Vitro Biological Agent to Biocontrol of Date Palm Stipe Rot Affected by Fusarium brachygibbosum by Using Pergularia tomentosa L., Aqueous Extract

Palm (Phoenix dactylifera L.) is a specie cultivated in Mauritania. The present study is focus on the potential of the aqueous extract of the Pergularia tomentosa L., as biological control agent against Fusarium brachygibbosum, the main agent of the date palm stipe rot disease in Mauritania. Fungal pathogen was isolated from symptomatic date palm stipe rot disease in Mauritania. The morphological characterization and the molecular identification by sequencing ITS1, ITS2 5.8 RNAr region showed homology of 98% with Fusarium brachygibbosum strain UOA/HCPF 16982 s during in vitro tests on leaves performed to verify its phytopathogenicity. the inhibitory effect of aqueous extract of Pergularia tomentosa L. on the phytopathogenic isolate (Fusarium brachygibbosum) of date palm, on mycelial growth and spore germination was observed. In fact, in vitro on PDA, mycelial growth ranged from 39.23 to 67.7 %, depending on the concentration of the aqueous extract of P. tomentosa. The aqueous extract showed a reduction of spore germination varying from 5.3 to 23.8 %. The minimum inhibitory concentrations varied from 1 to 100 mg/ml. the various extracts give high antioxidant activities exhibited by DPPH assay. Multivariant analysis by PCA plot and the heatmap were done, to evaluate the correlation between the tested parameters. These results suggest the use of Pergularia tomentosa L., extract as a biological agent to control and reduce damage caused by Fusarium brachygibbosum.National Agency for Scientific Research and Innovation in Mauritani

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.Compañía Logística de Hidrocarburos S.

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.Spanish GovernmentEuropean Research Founds CTM2017-84332-RJunta de Andalucia B-RNM-204-UGR20Consejo Nacional de Ciencia y Tecnologia (CONACyT) 377965 739637Spanish Ministry of Universities (Maria Zambrano Program) - Next Generation EU (NGEU

El escape room como técnica de estudio para la asignatura de Microbiología I

La motivación es un elemento fundamental para el aprendizaje del alumnado. Sin embargo, gran parte del contenido al que a veces se enfrentan no resulta motivante y genera una falta de implicación. En este sentido, las estrategias metodológicas deberán ir enfocadas estimular la motivación del alumnado. Una de las estrategias de innovación es la gamificación. Este enfoque se utiliza para aplicar los conocimientos adquiridos de manera lúdica en contextos que, a priori, no lo son. La gamificación es una estrategia didáctica que permite mejorar la motivación y, en consecuencia, la experiencia del aprendizaje haciéndolo más atractivo. El objetivo de este proyecto es aplicar la gamificación en la asignatura de Microbiología I del Grado de Farmacia como herramienta motivadora. Para ello, se llevará a cabo un escape room al final del primer cuatrimestre, una vez finalizado el contenido de la asignatura con fecha cercana la realización del examen final, dónde los alumnos previamente inscritos realicen esta prueba en grupos de 4 ó 5 personas con una duración de aproximadamente 20 minutos. El enigma del escape room irá enfocado a conceptos relativos a la asignatura, y se considerará como una actividad voluntaria dentro de la asignatura de Microbiología I. De este modo, se pretende usar la actividad del escape room como una herramienta de estudio previa al examen para afianzar y mejorar los conocimientos adquiridos durante la asignatura y la preparación por parte del estudiante para su evaluación en el examen final. Para valorar los efectos de esta actividad, se evaluará el rol de los alumnos durante el desarrollo de la misma, así como si ésta ha sido resuelta. También se analizará el impacto de esta actividad en la calificación del examen final. Los alumnos también valorarán esta actividad y una autoevaluación con respecto a su aprendizaje

Twitter as a Tool for Teaching and Communicating Microbiology: The #microMOOCSEM Initiative

Online social networks are increasingly used by the population on a daily basis. They are considered a powerful tool for science communication and their potential as educational tools is emerging. However, their usefulness in academic practice is still a matter of debate. Here, we present the results of our pioneering experience teaching a full Basic Microbiology course via Twitter (#microMOOCSEM), consisting of 28 lessons of 40-45 minutes duration each, at a tweet per minute rate during 10 weeks. Lessons were prepared by 30 different lecturers, covering most basic areas in Microbiology and some monographic topics of general interest (malaria, HIV, tuberculosis, etc.). Data analysis on the impact and acceptance of the course were largely affirmative, promoting a 330% enhancement in the followers and a >350-fold increase of the number of visits per month to the Twitter account of the host institution, the Spanish Society for Microbiology. Almost one third of the course followers were located overseas. Our study indicates that Massive Online Open Courses (MOOC) via Twitter are highly dynamic, interactive, and accessible to great audiences, providing a valuable tool for social learning and communicating science. This strategy attracts the interest of students towards particular topics in the field, efficiently complementing customary academic activities, especially in multidisciplinary areas like Microbiology.Versión del edito

Estudio de los procesos biológicos y de la estructura de las comunidades microbianas en el proceso de compostaje de lodos de depuradora de aguas residuales urbanas, en sistemas de membranas semipermea

El compostaje es un proceso viable para el tratamiento final de los lodos procedentes de la depuración del agua, en un contexto en el que la tendencia de este residuo tiende a incrementarse. Su uso final como enmienda en suelos representa una opción sostenible desde un punto de vista ambiental y económico y conforme con las nuevas políticas ambientales de reciclaje y revalorización de residuos. A pesar de existir numerosas tecnologías para el compostaje de lodos de depuradora, el empleo de cubiertas semipermeables representa una tecnología que permite el control de olores y el mantenimiento de la temperatura y humedad en el interior de la pila, lo que puede acelerar dicho proceso. Los microorganismos son los principales responsables, por lo que la monitorización de los cambios poblacionales y su interrelación con las variables operacionales puede representar una herramienta muy útil para optimizar estos procesos. En este trabajo se llevó a cabo el estudio de dos pilas de compostaje con lodos de depuradora a escala industrial. En ambas pilas se utilizó la tecnología de cubierta semipermeable conectada con un sistema de insuflación de aire, modificando únicamente los tiempos de compostaje. Se llevó a cabo la monitorización de los parámetros físico-químicos y microbiológicos, como la determinación de las actividades enzimáticas y el estudio de la diversidad microbiana (bacteriana, fúngica y vírica) tanto mediante técnicas dependientes como independientes de cultivo. Finalmente se realizó un cribado de la producción de enzimas extracelulares en los aislados bacterianos y fúngicos obtenidos con el fin de la obtención de enzimas de valor biotecnológico. Las variables físico-químicas revelaron una alta correlación entre la actividad microbiana y la diversidad, reflejadas en una intensa fase de degradación bajo la cubierta. La diversidad bacteriana y fúngica se vio influenciada principalmente por las altas temperaturas, que favorecieron a las comunidades termotolerantes. La comunidad vírica estuvo caracterizada por la presencia de virus de plantas y bacteriófagos de bacterias entéricas, sugiriendo el importante papel de los virus en el control de las comunidades patógenas.Composting is a viable process for the final treatment of sewage sludge, in a context in which these wastes tend to increase. The addition of compost in soil represents a sustainable option from an environmental and economic point of view. Likewise, this application is in agreement with new environmental policies of recycling and in the revaluation of waste. Despite the existence of numerous technologies for the composting of sewage sludge, the use of semipermeable covers represent a technology that allows the control of odors and the maintenance of temperature and humidity inside the system, which can accelerate the composting process. Microorganisms are the main responsible players of the biological transformation, thus, monitoring the changes in the microbial population and their interrelation with operational variables can represent a very useful tool to optimize these processes. In this work, we carried out the study of two composting piles with sewage sludge at industrial scale. Both piles used the semipermeable cover technology connected with an air insufflation system, and were tested under variable composting times. The monitored physical-chemical and microbiological parameters included the determination of enzymatic activities and the study of microbial diversity (bacterial, fungal and viral population). These variables were tracked by both dependent and independent culture technologies. Finally, the production of extracellular enzymes in bacterial and fungal isolates was studied to obtain enzymes of biotechnological value. Physico-chemical variables revealed high correlation between microbial activity and diversity, reflected in an intense phase of degradation under the semipermeable cover. The bacterial and fungal diversity was most influenced by the high temperatures, which favored thermotolerant communities. The viral community was characterized by the presence of plant viruses and bacteriophages of enteric bacteria. Results suggest the important role of viruses in the control of pathogenic communities.Tesis Univ. Granada.Este trabajo ha sido financiado por el Proyecto de Excelencia Motriz RNM 7370 de la Junta de Andalucía y se ha llevado a cabo dentro del Grupo RNM-270 de Microbiología Ambiental de la Universidad de Granada. La autora agradece el contrato predoctoral ofrecido por la Junta de Andalucía.Programa Oficial de Doctorado en Biología Fundamental y de Sistema

Pharmaceutical active compounds in sewage sludge: Degradation improvement and conversion into an organic amendment by composting processes

This work was supported by the Spanish Ministry of Economy and Competitiveness and European Research Funds (FEDER) [grant numbers CTM2017-84332-R] and from The Junta de Andalucía FEDER/Junta de Andalucía-Consejería de Transformación Económica, Industria, Conocimiento y Universidades [B-RNM-204-UGR20]. GAP studies were funded by a CONACyt fellowship [grant number 772485] the contract of TRM has been funded by María Zambrano Program (Next Generation Funds, UE) and SG received a Undergraduate Initiation to Research Scholarship from the University of Granada [grant number 669608935].Appendix A. Supplementary material Supplementary data to this article can be found online at https://doi. org/10.1016/j.wasman.2023.05.055.Around 143,000 chemicals find their fate in wastewater treatment plants in the European Union. Low efficiency on their removal at lab-based studies and even poorer performance at large scale experiments have been reported. Here, a coupled biological technology (bioaugmentation and composting) is proposed and proved for pharmaceutical active compounds degradation and toxicity reduction. The optimization was conducted through in situ inoculation of Penicillium oxalicum XD 3.1 and an enriched consortium (obtained from non-digested sewage sludge), into pilot scale piles of sewage sludge under real conditions. This bioaugmentation-composting system allowed a better performance of micropollutants degradation (21 % from the total pharmaceuticals detected at the beginning of the experiment) than a traditional composting process. Particularly, inoculation with P. oxalicum allowed the degradation of some recalcitrant compounds like carbamazepine, cotinine and methadone, and also produced better stabilization features in the mature compost (significant passivation of copper and zinc, higher macronutrients value, adequate physicochemical conditions for soil direct application and less toxic effect on germination) compared to the control and the enriched culture. These findings provide a feasible, alternative strategy to obtain a safer mature compost and a better removal of micropollutants performance at large scale.Grupo RNM-270 UGRSpanish Ministry of Economy and Competitiveness and European Research Funds (FEDER) [grant numbers CTM2017-84332-R]Junta de Andalucía FEDER/Junta de Andalucía-Consejería de Transformación´Económica, Industria, Conocimiento y Universidades [B-RNM-204-UGR20]CONACyt fellowship [grant number 772485]María Zambrano Program (Next Generation Funds, UE)Undergraduate Initiation to Research Scholarship from the University of Granada [grant number 669608935

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

First Report of in vitro Biological Agent to Biocontrol of Date Palm Stipe Rot Affected by Fusarium brachygibbosum by Using Pergularia tomentosa L., Aqueous Extract

Palm (Phoenix dactylifera L.) is a specie cultivated in Mauritania. The present study is focus on the potential of the aqueous extract of the Pergularia tomentosa L., as biological control agent against Fusarium brachygibbosum, the main agent of the date palm stipe rot disease in Mauritania. Fungal pathogen was isolated from symptomatic date palm stipe rot disease in Mauritania. The morphological characterization and the molecular identification by sequencing ITS1, ITS2 5.8 RNAr region showed homology of 98% with Fusarium brachygibbosum strain UOA/HCPF 16982 s during in vitro tests on leaves performed to verify its phytopathogenicity. the inhibitory effect of aqueous extract of Pergularia tomentosa L. on the phytopathogenic isolate (Fusarium brachygibbosum) of date palm, on mycelial growth and spore germination was observed. In fact, in vitro on PDA, mycelial growth ranged from 39.23 to 67.7 %, depending on the concentration of the aqueous extract of P. tomentosa. The aqueous extract showed a reduction of spore germination varying from 5.3 to 23.8 %. The minimum inhibitory concentrations varied from 1 to 100 mg/ml. the various extracts give high antioxidant activities exhibited by DPPH assay. Multivariant analysis by PCA plot and the heatmap were done, to evaluate the correlation between the tested parameters. These results suggest the use of Pergularia tomentosa L., extract as a biological agent to control and reduce damage caused by Fusarium brachygibbosum