Antibiotic Resistance in Agricultural Soil and Crops Associated to the Application of Cow Manure-Derived Amendments from Conventional and Organic Livestock Farms

he application of organic amendments to agricultural soil can enhance crop yield, while improving the physicochemical and biological properties of the recipient soils. However, the use of manure-derived amendments as fertilizers entails environmental risks, such as the contamination of soil and crops with antibiotic residues, antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs). In order to delve into these risks, we applied dairy cow manure-derived amendments (slurry, fresh manure, aged manure), obtained from a conventional and an organic farm, to soil. Subsequently, lettuce and wheat plants were grown in the amended soils. After harvest, the abundance of 95 ARGs and MGE-genes from the amended soils and plants were determined by high-throughput qPCR. The structure of soil prokaryotic communities was determined by 16S rRNA amplicon sequencing and qPCR. The absolute abundance of ARGs and MGE-genes differed between treatments (amended vs. unamended), origins of amendment (conventional vs. organic), and types of amendment (slurry vs. fresh manure vs. aged manure). Regarding ARG-absolute abundances in the amendments themselves, higher values were usually found in slurry vs. fresh or aged manure. These abundances were generally higher in soil than in plant samples, and higher in wheat grain than in lettuce plants. Lettuce plants fertilized with conventional amendments showed higher absolute abundances of tetracycline resistance genes, compared to those amended with organic amendments. No single treatment could be identified as the best or worst treatment regarding the risk of antibiotic resistance in soil and plant samples. Within the same treatment, the resistome risk differed between the amendment, the amended soil and, finally, the crop. In other words, according to our data, the resistome risk in manure-amended crops cannot be directly inferred from the analysis of the amendments themselves. We concluded that, depending on the specific question under study, the analysis of the resistome risk should specifically focus on the amendment, the amended soil or the cropThis work has been financially supported by the Basque Government (projects: URAGAN and KONTRAE-Elkartek-KK-2020-00007) and the Spanish Ministry of Science and Innovation (project: PRADA PID2019-110055RB-C21). LJ was the recipient of a predoctoral fellowship from the Department of Economic Development and Infrastructure of the Basque Governmen

Agricultural Soils Amended with Thermally-Dried Anaerobically-Digested Sewage Sludge Showed Increased Risk of Antibiotic Resistance Dissemination

The application of sewage sludge (SS) to agricultural soil can help meet crop nutrient requirements and enhance soil properties, while reusing an organic by-product. However, SS can be a source of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs), resulting in an increased risk of antibiotic resistance dissemination. We studied the effect of the application of thermally-dried anaerobically-digested SS on (i) soil physicochemical and microbial properties, and (ii) the relative abundance of 85 ARGs and 10 MGE-genes in soil. Soil samples were taken from a variety of SS-amended agricultural fields differing in three factors: dose of application, dosage of application, and elapsed time after the last application. The relative abundance of both ARGs and MGE-genes was higher in SS-amended soils, compared to non-amended soils, particularly in those with a more recent SS application. Some physicochemical parameters (i.e., cation exchange capacity, copper concentration, phosphorus content) were positively correlated with the relative abundance of ARGs and MGE-genes. Sewage sludge application was the key factor to explain the distribution pattern of ARGs and MGE-genes. The 30 most abundant families within the soil prokaryotic community accounted for 66% of the total variation of ARG and MGE-gene relative abundances. Soil prokaryotic alpha-diversity was negatively correlated with the relative abundance of ARGs and MGE-genes. We concluded that agricultural soils amended with thermally-dried anaerobically-digested sewage sludge showed increased risk of antibiotic resistance dissemination.This work has been financially supported by the Basque Government (projects: URAGAN and KONTRAE-Elkartek-KK2020-00007) and the Spanish Ministry of Science and Innovation (project: PRADA PID2019-110055RB-C21). LJ was the recipient of a predoctoral fellowship from the Department of Economic Development and Infrastructure of the Basque Government

Organic amendment treatments for antimicrobial resistance and mobile element genes risk reduction in soil-crop systems

Abstract Agricultural fertilization with organic amendments of animal origin often leads to antibiotic resistance dissemination. In this study, we evaluated the effect of different treatments (anaerobic digestion, biochar application, ozonation, zerovalent iron nanoparticle application, and spent mushroom substrate addition) on the resistome in dairy cow manure-derived amendments (slurry, manure, and compost). Anaerobic digestion and biochar application resulted in the highest reduction in antibiotic resistance gene (ARG) and mobile genetic element (MGE) gene abundance. These two treatments were applied to cow manure compost, which was then used to fertilize the soil for lettuce growth. After crop harvest, ARG and MGE gene absolute and relative abundances in the soil and lettuce samples were determined by droplet digital PCR and high-throughput qPCR, respectively. Prokaryotic diversity in cow manure-amended soils was determined using 16S rRNA metabarcoding. Compared to untreated compost, anaerobic digestion led to a 38% and 83% reduction in sul2 and intl1 absolute abundances in the soil, respectively, while biochar led to a 60% reduction in intl1 absolute abundance. No differences in lettuce gene abundances were observed among treatments. We conclude that amendment treatments can minimize the risk of antibiotic resistance in agroecosystems

La innovación escolar desde la perspectiva de personas con enfermedades raras en el País Vasco: historias de vida, prácticas escolares, necesidades del sistema educativo y propuestas de mejora para una escuela y sociedad inclusiva

140 p.[ES] En este trabajo se presenta la investigación “La innovación escolar desde la perspectiva de personas con enfermedades raras en el País Vasco” en el que un grupo de jóvenes con enfermedades raras y sus familias comparten sus experiencias de escolarización para avanzar en los procesos de innovación y mejora escolar en el marco de una escuela inclusiva como respuesta a los retos y necesidades de nuestra sociedad.[EN] In this paper we present a research work titled “School innovation from the perspective of people with rare diseases in the Basque Country”. The methodology is based on the voice and the life stories of pupils with rare diseases and their families. They share their experiences of schooling to progress in the innovation processes and school improvement. We also present an option for an inclusive school in response to the challenges and needs of our society.[EU] Lan honetan aurkezten da "Eskola berrikuntza gaixotasun arraroak dituzten pertsonen ikuspegitik Euskal Autonomi Erkidegoan" izeneko ikerketa. Bertan gaixotasun arraroak dituzten gazte talde batek eta euren familiek euren eskolatze esperientziak kontatzen dituzte; berrikuntza eta eskola hobekuntzan aurrerapausoak emateko asmoz. Eskola inklusiboaren aldeko aukera aurkezten da, egun ditugun gizarte honetako erronka eta beharrei aurre egiteko.Patrocina Fundación Mikel Uriart

La innovación escolar desde la perspectiva de personas con enfermedades raras en el País Vasco: historias de vida, prácticas escolares, necesidades del sistema educativo y propuestas de mejora para una escuela y sociedad inclusiva

140 p.[ES] En este trabajo se presenta la investigación “La innovación escolar desde la perspectiva de personas con enfermedades raras en el País Vasco” en el que un grupo de jóvenes con enfermedades raras y sus familias comparten sus experiencias de escolarización para avanzar en los procesos de innovación y mejora escolar en el marco de una escuela inclusiva como respuesta a los retos y necesidades de nuestra sociedad.[EN] In this paper we present a research work titled “School innovation from the perspective of people with rare diseases in the Basque Country”. The methodology is based on the voice and the life stories of pupils with rare diseases and their families. They share their experiences of schooling to progress in the innovation processes and school improvement. We also present an option for an inclusive school in response to the challenges and needs of our society.[EU] Lan honetan aurkezten da "Eskola berrikuntza gaixotasun arraroak dituzten pertsonen ikuspegitik Euskal Autonomi Erkidegoan" izeneko ikerketa. Bertan gaixotasun arraroak dituzten gazte talde batek eta euren familiek euren eskolatze esperientziak kontatzen dituzte; berrikuntza eta eskola hobekuntzan aurrerapausoak emateko asmoz. Eskola inklusiboaren aldeko aukera aurkezten da, egun ditugun gizarte honetako erronka eta beharrei aurre egiteko.Patrocina Fundación Mikel Uriart

Genetic and epigenetic modifications of Sox2 contribute to the invasive phenotype of malignant gliomas.

We undertook this study to understand how the transcription factor Sox2 contributes to the malignant phenotype of glioblastoma multiforme (GBM), the most aggressive primary brain tumor. We initially looked for unbalanced genomic rearrangements in the Sox2 locus in 42 GBM samples and found that Sox2 was amplified in 11.5% and overexpressed in all the samples. These results prompted us to further investigate the mechanisms involved in Sox2 overexpression in GBM. We analyzed the methylation status of the Sox2 promoter because high CpG density promoters are associated with key developmental genes. The Sox2 promoter presented a CpG island that was hypomethylated in all the patient samples when compared to normal cell lines. Treatment of Sox2-negative glioma cell lines with 5-azacitidine resulted in the re-expression of Sox2 and in a change in the methylation status of the Sox2 promoter. We further confirmed these results by analyzing data from GBM cases generated by The Cancer Genome Atlas project. We observed Sox2 overexpression (86%; N = 414), Sox2 gene amplification (8.5%; N = 492), and Sox 2 promoter hypomethylation (100%; N = 258), suggesting the relevance of this factor in the malignant phenotype of GBMs. To further explore the role of Sox2, we performed in vitro analysis with brain tumor stem cells (BTSCs) and established glioma cell lines. Downmodulation of Sox2 in BTSCs resulted in the loss of their self-renewal properties. Surprisingly, ectopic expression of Sox2 in established glioma cells was not sufficient to support self-renewal, suggesting that additional factors are required. Furthermore, we observed that ectopic Sox2 expression was sufficient to induce invasion and migration of glioma cells, and knockdown experiments demonstrated that Sox2 was essential for maintaining these properties. Altogether, our data underscore the importance of a pleiotropic role of Sox2 and suggest that it could be used as a therapeutic target in GBM