Metabolic interactions within microbial communities are essential for the efficient degradation of complex organic compounds, and underpin natural phenomena driven by microorganisms, such as the recycling of carbon-, nitrogen-, and sulfur-containing molecules. These metabolic interactions ultimately determine the function, activity and stability of the community, and therefore their understanding would be essential to steer processes where microbial communities are involved. This is exploited in the design of microbial fuel cells (MFCs), bioelectrochemical devices that convert the chemical energy present in substrates into electrical energy through the metabolic activity of microorganisms, either single species or communities. In this work, we analyzed the evolution of the microbial community structure in a cascade of MFCs inoculated with an anaerobic microbial community and continuously fed with a complex medium. The analysis of the composition of the anodic communities revealed the establishment of different communities in the anodes of the hydraulically connected MFCs, with a decrease in the abundance of fermentative taxa and a concurrent increase in respiratory taxa along the cascade. The analysis of the metabolites in the anodic suspension showed a metabolic shift between the first and last MFC, confirming the segregation of the anodic communities. Those results suggest a metabolic interaction mechanism between the predominant fermentative bacteria at the first stages of the cascade and the anaerobic respiratory electrogenic population in the latter stages, which is reflected in the observed increase in power output. We show that our experimental system represents an ideal platform for optimization of processes where the degradation of complex substrates is involved, as well as a potential tool for the study of metabolic interactions in complex microbial communities.

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The rapid emergence of antibiotic resistant bacterial pathogens constitutes a critical problem in healthcare and requires the development of novel treatments. Potential strategies include the exploitation of microbial social interactions based on public goods, which are produced at a fitness cost by cooperative microorganisms, but can be exploited by cheaters that do not produce these goods. Cheater invasion has been proposed as a 'Trojan horse' approach to infiltrate pathogen populations with strains deploying built-in weaknesses (e.g. sensitiveness to antibiotics). However, previous attempts have been often unsuccessful because population invasion by cheaters was prevented by various mechanisms including the presence of spatial structure (e.g. growth in biofilms), which limits the diffusion and exploitation of public goods.Here we followed an alternative approach and examined whether the manipulation of public good uptake and not its production could result in potential 'Trojan horses' suitable for population invasion. We focused on the siderophore pyoverdine produced by the human pathogen Pseudomonas aeruginosa MPAO1 and manipulated its uptake by deleting and/or overexpressing the pyoverdine primary (FpvA) and secondary (FpvB) receptors. We found that receptor synthesis feeds back on pyoverdine production and uptake rates, which led to strains with altered pyoverdine-associated costs and benefits. Moreover, we found that the receptor FpvB was advantageous under iron-limited conditions but revealed hidden costs in the presence of an antibiotic stressor (gentamicin). As a consequence, FpvB mutants became the fittest strain under gentamicin exposure, displacing the wildtype in liquid cultures, and in biofilms and during infections of the wax moth larvae Galleria mellonella, which both represent structured environments. Our findings reveal that an evolutionary trade-off associated with the costs and benefits of a versatile pyoverdine uptake strategy can be harnessed for devising a Trojan horse candidate for medical interventions.

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The clinical and industrial importance of clavulanic acid (CA) promotes the search for novel strategies to boost productivity at lower costs. In this contribution, the production of CA was evaluated in two reference strains of Streptomyces clavuligerus (Sc), ATCC27064 and DSM41826; the strains were tested in different culture media, using product yields and culture media costs as selection criteria. On average, ScATCC reached a concentration of CA 1.6-fold higher than ScDSM, using the isolated soybean protein (ISP) culture medium. A central composite experimental design was used to optimize the medium, by studying the influence of ISP and K2HPO4 concentration on the production of CA. The maximum CA concentration obtained in this optimized medium was 56 mg·L -1 , close to the theoretical optimum. Our results confirm that a culture medium containing a rich carbon source such as glycerol and/or starch provides the best conditions for CA production.Keywords: clavulanic acid; culture media design; strain selection; Streptomyces clavuligerus; antibiotics.Selección de una cepa de Streptomyces clavuligerus para la biosíntesis de ácido clavulánico: un estudio basado en los efectos de la composición del medio de cultivo y análisis estadístico Resumen La importancia clínica e industrial del ácido clavulánico (AC) promueve la búsqueda de nuevas estrategias para aumentar su productividad a un menor costo. En esta contribución, se evaluó la producción de AC en dos cepas de referencia de Streptomyces clavuligerus (Sc), ATCC27064 y DSM41826; éstas se probaron en diferentes medios de cultivo; los rendimientos de producto y costo de los medios se usaron como criterios de selección. En promedio, ScATCC alcanzó una concentración de AC 1.6-veces mayor que ScDSM, en el medio de proteína de soya (ISP). Se optimizó el medio con un diseño experimental central compuesto, estudiando la influencia del ISP y K2HPO4 sobre la producción de AC. La concentración máxima de AC obtenida en el medio optimizado (56mg·L -1 ) estuvo cerca del óptimo teórico. Nuestros resultados confirman que medios de cultivo con fuentes de carbono como glicerol y/o almidón proporcionan las mejores condiciones para la producción de AC.Palabras clave: ácido clavulánico; diseño de medios de cultivo; selección de cepa; Streptomyces clavuligerus; antibióticos.