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Resumen Habiendo detectado la necesidad de actualizar las técnicas docentes empleadas por el profesorado universitario, estamos desarrollando, en colaboración con el Departamento de Ciencias Naturales del Colegio Cristo Rey de Jaén, el proyecto titulado “Transferencia de prácticas docentes de trabajo cooperativo del ámbito pre-universitario a la docencia práctica de asignaturas de Grado y Postgrado en la Universidad de Jaén”, concedido en la Convocatoria de Proyectos de Innovación e Incentivación de las Buenas Prácticas Docentes en la Universidad de Jaén y que implica a cuatro áreas de la Facultad de Ciencias Experimentales. En concreto en el área de Microbiología hemos desarrollado un bloque de prácticas mediante la técnica de Aprendizaje Basado en Problemas y el resto mediante la metodología tradicional de explicación previa del profesor y desarrollo de la práctica. Tanto la motivación del profesorado como de los estudiantes ha mejorado tras la aplicación de esta técnica docente. La adquisición de competencias también ha sido superior en las prácticas desarrolladas con esta metodología, en comparación con las desarrolladas por el método tradicional. Concluimos que esta metodología docente se puede adaptar perfectamente a la enseñanza universitaria, puede facilitar la comprensión del fundamento teórico y la adquisición de competencias en algunas clases prácticas, además de mejorar la motivación tanto del profesorado como de los estudiantes. Abstract Pre-university teachers have probed experience in applying new and innovative teaching methods, so we have been guided by professors from the High School “Cristo Rey Jaén” to develop these techniques in the scope of applicability of a Teaching Innovation Project titled “Transfer of collaborative teaching techniques from pre-university levels to practical classes at the University of Jaén”, that we obtained from this University and that implies the joint effort of four areas of knowledge in the Faculty of Experimental Sciences at the University of Jaén. Problem-Based Learning (PBL), a pedagogical approach and curriculum design methodology often used in higher education, has been used in practical classes in the Microbiology Unit. We have detected higher drive in both, teachers and students in those practical classes conducted with this methodology, when compared to traditional techniques. Proficiency acquired by students was also higher when applying this methodology, so we conclude it can be perfectly transposed to University in order to improve the understanding of the theoretical basis of practical classes, the competence acquisition by students and to increase the motivation of both, teachers and students.

Copper-derived compounds are often used in olive tree farms. In a previous study, a collection of bacterial strains isolated from olive tree farms were identified and tested for phenotypic antimicrobial resistance and heavy metal tolerance. The aim of this work was to study the genetic determinants of resistance and to evaluate the co-occurrence of metal tolerance and antibiotic resistance genes. Both metal tolerance and antibiotic resistance genes (including beta-lactamase genes) were detected in the bacterial strains from Cu-treated soils. A high percentage of the strains positive for metal tolerance genes also carried antibiotic resistance genes, especially for genes involved in resistances to beta-lactams and tetracycline. Significant associations were detected between genes involved in copper tolerance and genes coding for beta-lactamases or tetracycline resistance mechanisms. A significant association was also detected between zntA (coding for a Zn(II)-translocating P-type ATPase) and tetC genes. In conclusion, bacteria from soils of Cu-treated olive farms may carry both metal tolerance and antibiotic resistance genes. The positive associations detected between metal tolerance genes and antibiotic resistance genes suggests co-selection of such genetic traits by exposure to metals.

Natural A-type procyanidins have shown very interesting biological activities, such as their proven antiadherence properties against pathogenic bacteria. In order to find the structural features responsible for their activities, we describe herein the design and synthesis of six A-type procyanidin analogues and the evaluation of their antimicrobial and antibiofilm properties against 12 resistant bacteria, both Gram positive and Gram negative, isolated from organic foods. The natural A-type procyanidin A-2, which had known antiadherence activity, was also tested as a reference compound for the comparative studies. Within the series, analogue 4, which had a NO2 group on ring A, showed the highest antimicrobial activity (MIC of 10 μg/mL) and was one of the best molecules at preventing biofilm formation (up to 40% decreases at 100 μg/mL) and disrupting preformed biofilms (up to 40% reductions at 0.1 μg/mL). Structure–activity relationships are also analyzed.

A collection of 38 biocide-adapted strains with significant increases in their tolerance to biocides after step-wise exposure to these compounds were screened for sensitivity to essential oils and chemical preservatives. Several strains adapted to quaternary ammonium compounds (benzalkonium chloride, hexadecylpyridinium chloride or cetrimide) showed a generalized increase in the sensitivity to preservatives. Similar results were found among hexachlorophene- or chlorhexidine-adapted strains. Moreover, tolerance to hexadecylpyridinium chloride showed a very strong positive correlation with 4-hydroxybenzoic acid, thyme oil and sodium nitrite increased sensitivity, as well as a strong correlation with clove oil, potassium sorbate and potassium nitrate increased sensitivity. On the contrary, an increase in tolerance to preservatives was detected among triclosan-adapted strains. Results from this study suggest that exposure of bacteria from foods to biocides is not always associated with co-selection for other antimicrobial resistances, especially against essential oils or chemical preservatives used in the food industry.

Summary This study aimed to evaluate the antimicrobial and antibiofilm activities of two procyanidins isolated from an ethyl acetate extract of laurel wood against a selection of foodborne pathogens. The analysis of the extract by HPLC–DAD/ESI–MS allowed us to detect the presence of two procyanidins, which were selectively isolated and identified by chromatographic and spectroscopic means as cinnamtannin B-1 (1) and procyanidin B-2 (2). Procyanidins 1 and 2 exhibited two biological activities: inhibition of bacterial growth at high concentrations and prevention of biofilm formation at lower concentrations. Synergistic effect was also detected when both compounds were tested in combination against Listeria monocytogenes. Significant effects were also detected on disruption of preformed biofilm. The ability of procyanidins to inhibit microbial growth and biofilm formation and to synergistically work with each other may stimulate a market as natural food preservatives, and/or natural sanitisers for processing equipment where foodborne pathogens reside.

Background Heavy metal pollution may act as persistent selective pressure that favors the spread of antimicrobial resistance in natural environments. The aim of this study was to isolate and identify metal-tolerant bacteria from soils in olive tree fields routinely treated with copper-derived compounds and to evaluate the tolerance of bacterial strains to other metals and their resistance to clinically relevant antibiotics. Results Five hundred and ninety-five bacterial isolates from 45 olive tree agricultural fields were studied. Minimum inhibitory concentrations (MICs) ≥ 16 mmol L-1 were detected for copper (57% of isolates), zinc (37%) and lead (62%), while only 3% had MICs ≥ 12 mmol L-1 for nickel. Ninety-six metal-tolerant strains were selected for identification and antibiotic resistance determination. Most isolates belonged to the genera Pseudomonas (37%), Bacillus (23%) and Chryseobacterium (20%), while 6% were identified as Variovorax, 4% as Stenotrophomonas and 2% as Serratia or Burkholderia. Highest copper tolerance was detected among Pseudomonas. Over 75% of the strains with high copper tolerance were also resistant to vancomycin, 50% to ampicillin and 40% to erythromycin or trimethoprim/sulfamethoxazole. Conclusion Bacteria from olive soils are tolerant to metals, mainly copper, but also zinc and lead, as well as resistant to clinically important antibiotics, which could be a troublesome issue in clinical settings. © 2019 Society of Chemical Industry.

Heterofermentative lactic acid bacteria (76 strains) belonging to Lactobacillus, Leuconostoc and Weissella species which are important in fermentation, spoilage or as probiotics were screened in a factorial experiment for their ability to grow, produce catalase and consume oxygen in aerobiosis or in anaerobiosis, with or without supplementation with hemin and/or menaquinone in a medium containing glucose as a carbohydrate source. Aerobiosis improved growth with a few exceptions. The effect of supplementation with heme and/or menaquinone was strain specific and clear evidence of heme-boosted respiration was found in some cases. Heme-catalase was produced by strains of L. brevis, W. minor and Leuc. mesenteroides; some strains of the latter species produced non-heme catalase. Shaken flasks experiments showed that aerobic growth resulted in increased maximum growth rate and in a limited increase in biomass. Heme supplementation during aerobic growth resulted in a further increase in growth rate and final biomass only for a few strains; this was often related to catalase, which was also responsible for increased tolerance of H2O2. In both experiments we found evidence of heme toxicity, especially in anaerobiosis and in absence of menaquinone. Dose response curves for aerobic growth in the presence of combinations of hemin and menaquinone were non-monotonic, with growth stimulation at low doses of heme (

Chlorhexidine (CH) and quaternary ammonium compounds (QAC), such as cetrimide (CE), are widely used as disinfectants because of their broad antimicrobial spectrum. However, their frequent use for disinfection in different settings may promote bacterial drug resistance against both biocides and clinically relevant antibiotics. This study analyzes the effects of stepwise exposure to cetrimide (CE) and chlorhexidine (CH) of bacteria from organic foods and previously classified as biocide-sensitive. Gradual exposure of these strains to biocides resulted in mainly transient decreased antimicrobial susceptibility to other antibiotics and to biocides. Biocide-adapted bacteria also exhibit alterations in physiological characteristics, mainly decreased heat tolerance, or gastric acid tolerance in CE-adapted strains, while bile resistance does not seem to be influenced by biocide adaptation. Results from this study suggest that changes in membrane fluidity may be the main mechanism responsible for the acquisition of stable tolerance to biocides.