Mucoid Coating Provides a Growth Advantage to Pseudomonas aeruginosa at Oil-Water Interfaces.

Chronic lung infection with bacterial biofilms is one of the leading causes of death in cystic fibrosis (CF) patients. Among many species infecting the lung airways, Pseudomonas aeruginosa is the major pathogen colonizing and persisting throughout the patient's life. The microorganism undergoes pathoadaptation, while switching from a nonmucoid to a mucoid phenotype, improving the mechanical properties of the resulting biofilms. Previous investigation of the dynamic rheological properties of nonmucoid (PANT) and mucoid (PASL) clinical P. aeruginosa isolates exposed to interfacial stresses demonstrated that the mucoid strains formed films with stronger resistance to bending and nonlinear relaxation to compression and tension. We hypothesize that the mucoid switch provides a growth advantage to P. aeruginosa through the development of interfacial films with viscoelastic properties enabling cell survival. Here, we investigate the physiological response of the mucoid and the nonmucoid P. aeruginosa to interfacial entrapment. Our results, both macroscopic and molecular, reveal that mucoid coating plays an important role in protecting the bacteria from interfacial stresses. Cell characterizations using electron and fluorescence microscopies showed higher proportion of dead nonmucoid cells compared to mucoid cells on interfacial exposure. For example, scanning transmission electron microscopy (STEM) imaging showed that 96.6% of nonmucoid cells vs only 22.2% of mucoid cells were lysed owing to interfacial stress. Furthermore, the transcriptional profiling of P. aeruginosa cells indicated the upregulation of pel, psl, and alginate genes encoding for exopolysaccharide biomaterials is associated with mucoid cells' ability to cope with the interfacial environments. Further characterization of real-time gene regulation at interfaces will elucidate the effects of interfacial environment on the regulation of bacterial virulence.

Droplet-based microsystems as novel assessment tools for oral microbial dynamics.

The human microbiome comprises thousands of microbial species that live in and on the body and play critical roles in human health and disease. Recent findings on the interplay among members of the oral microbiome, defined by a personalized set of microorganisms, have elucidated the role of bacteria and yeasts in oral health and diseases including dental caries, halitosis, and periodontal infections. However, the majority of these studies rely on traditional culturing methods which are limited in their ability of replicating the oral microenvironment, and therefore fail to evaluate key microbial interactions in microbiome dynamics. Novel culturing methods have emerged to address this shortcoming. Here, we reviewed the potential of droplet-based microfluidics as an alternative approach for culturing microorganisms and assessing the oral microbiome dynamics. We discussed the state of the art and recent progress in the field of oral microbiology. Although at its infancy, droplet-based microtechnology presents an interesting potential for elucidating oral microbial dynamics and pathophysiology. We highlight how new findings provided by current microfluidic-based methodologies could advance the investigation of the oral microbiome. We anticipate that our work involving the droplet-based microfluidic technique with a semipermeable membrane will lay the foundations for future microbial dynamics studies and further expand the knowledge of the oral microbiome and its implication in oral health.

A systematic comparison of ectoine production from upgraded biogas using Methylomicrobium alcaliphilum and a mixed haloalkaliphilic consortium.

Biogas is the byproduct of anaerobic digestion with the highest valorization potential, however its full exploitation is limited by the lack of tax incentives and the inherent presence of pollutants. The development of technologies for biogas conversion into added-value products is crucial in order to ensure the competitiveness of this bioresource. This study constitutes the first proof of concept of upgraded biogas bioconversion into the high profit margin product ectoine. Ectoine represents the most expensive product synthesized by microorganisms with a retail value of 1000 $ kg-1 and a yearly increasing demand that currently entails a total market opportunity of 15000 M€. First, the production of ectoine from upgraded biogas was assessed in batch bioreactors. The presence of H2S did not exert a negative effect on the growth of the haloalkaliphilic ectoine producers, and ectoine yields up to 49 mg g biomass-1 were obtained. A second experiment conducted in continuous bubble column bioreactors confirmed the feasibility of the process under continuous mode (with ectoine yields of 109 mg g biomass-1). Finally, this study revealed that the removal of toxic compounds (i.e. medium dilution rate of 0.5 day-1) and process operation with a consortium composed of methylotrophic/non-methylotrophic ectoine producers enhanced upgraded biogas bioconversion. This research discloses the basis for the application of this innovative technology and could boost the economic performance of anaerobic digestion.

Biochar does not attenuate triclosan's impact on soil bacterial communities.

Triclosan, a broad-spectrum antimicrobial, has been widely used in pharmaceutical and personal care products. It undergoes limited degradation during wastewater treatment and is present in biosolids, most of which are land applied in the United States. This study assessed the impact of triclosan (0-100 mg kg-1) with and without biochar on soil bacterial communities. Very little 14C-triclosan was mineralized to 14CO2 (<7%) over the course of the study (42 days). While biochar (1%) significantly lowered mineralization of triclosan, analysis of 16S rRNA gene sequences revealed that biochar impacted very few OTUs and did not alter the overall structure of the community. Triclosan, on the other hand, significantly affected bacterial diversity and community structure (alpha diversity, ANOVA, p < 0.001; beta diversity, AMOVA, p < 0.01). Dirichlet multinomial mixtures (DMM) modeling and complete linkage clustering (CLC) revealed a dose-dependent impact of triclosan. Non-Parametric Metastats (NPM) analysis showed that 150 of 734 OTUs from seven main phyla were significantly impacted by triclosan (adjusted p < 0.05). Genera harboring opportunistic pathogens such as Flavobacterium were enriched in the presence of triclosan, as was Stenotrophomonas. The latter has previously been implicated in triclosan degradation via stable isotope probing. Surprisingly, Sphingomonads, which include well-characterized triclosan degraders were negatively impacted by even low doses of triclosan. Analyses of published genomes showed that triclosan resistance determinants were rare in Sphingomonads which may explain why they were negatively impacted by triclosan in our soil.