bConstitutive overproduction of the pump MexXY-OprM is recognized as a major cause of resistance to aminoglycosides, fluoroquinolones, and zwitterionic cephalosporins in Pseudomonas aeruginosa. In this study, 57 clonally unrelated strains recovered from non-cystic fibrosis patients were analyzed to characterize the mutations resulting in upregulation of the mexXY operon.

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After characterization of the reactive skin microbiota, we investigated whether the active Halymenia durvillei (HD), rich in polysaccharides, could modulate this microbiota after 28 days of treatment, act on neuroinflammation parameters, and calm feelings of discomfort and redness. Skin microbiota was assessed using next-generation sequencing experiments (16S RNA gene fragment sequencing) on samples collected from 30 volunteers suffering from reactive, sensitive skin. To evaluate the effect of the HD extract on neuroinflammation, we used an ex vivo model. Finally, an in vivo study was performed using a clinical assessment (blood microcirculation via videocapillaroscopy) of functional signs employing the Sensitive Scale and the soothing effect was evaluated and compared to a placebo treatment. At the phylum level, the samples were mostly composed of Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes, which accounted for more than 97% of the total sequencing read in all samples, with no differences before or after treatment with the HD active ingredient. The Shannon Diversity index indicated lower microbial communities compared to healthy skin. Maintenance of the Shannon Diversity index was reported after 28 days of HD active ingredient treatment, wherein microbial communities continued to decrease in number during treatment with the placebo. The average taxonomic composition of associated skin microbial communities showed that reactive skin is characterized by a low proportion of the Chryseobacterium genus compared to a high proportion of the Corynebacterium genus. At the species level, Actinobacteria are mainly represented by Propionibacterium acnes (72.13%) and Corynebacterium kroppenstedtii (13.23%), representing species typically observed in clinical cases of redness, the main criteria for volunteer inclusion. Corynebacterium kroppenstedtii, with increased levels being associated with skin redness, decreased with HD treatment. This decrease coincided with the clinical improvement observed after 7 weeks of treatment. The ex vivo study revealed that the HD extract induced a significant decrease in the expression of TRPV-1 (−67%; p < 0.001) and NK1-R (−43%; p < 0.01) compared to the control after 6 days of treatment. These data support the use of polysaccharides, found in red alga, in the treatment of reactive and sensitive skin related to the modulation of skin microbiota.

Pseudomonas aeruginosa is a human opportunistic pathogen responsible for nosocomial infections, which is largely used as a model organism to study antibiotic resistance and pathogenesis. As other species of the genus, its wide metabolic versatility appears to be attractive to study biotechnological applications. However, its natural resistance to antibiotics and its capacity to produce a wide range of virulence factors argue against its biotechnological potential. By reducing the genome of the reference strain PAO1, we explored the development of four hypovirulent and hypersusceptible recombinant DNA hosts (rDNA hosts). Despite deleting up to 0.8% of the core genome, any of the developed strains presented alterations of fitness when cultured under standard laboratory conditions. Other features such as antibiotic susceptibility, cytotoxicity, in vivo pathogenesis, and expression of heterologous peptides were also explored to highlight the potential applications of these models. This work stands as the first stage of the development of a safe-platform strain of Pseudomonas aeruginosa that will be further optimized for biotechnological applications.