Pseudomonas is a ubiquitous genus that also causes human, animal and plant diseases. Most studies have focused on clinical P. aeruginosa strains from humans, but they are scarce on animal strains. This study was aimed to determine the occurrence of Pseudomonas spp. among faecal samples of healthy animals, and to analyse their antimicrobial resistance, and pathogenicity. Among 704 animal faecal samples analysed, 133 Pseudomonas spp. isolates (23 species) were recovered from 46 samples (6.5%), and classified in 75 different PFGE patterns. Low antimicrobial resistance levels were found, being the highest to aztreonam (50.3%). Five sequence-types (ST1648, ST1711, ST2096, ST2194, ST2252), two serotypes (O:3, O:6), and three virulotypes (analysing 15 virulence and quorum-sensing genes) were observed among the 9 P. aeruginosa strains. Type-3-Secretion System genes were absent in the six O:3-serotype strains that additionally showed high cytotoxicity and produced higher biofilm biomass, phenazine pigments and motility than PAO1 control strain. In these six strains, the exlAB locus, and other virulence genotypes (e.g. RGP69 pathogenicity island) exclusive of PA7 outliers were detected by whole genome sequencing. This is the first description of the presence of the ExlA exolysin in P. aeruginosa from healthy animals, highlighting their pathological importance.

Mask usage has increased over the last few years due to the COVID-19 pandemic, resulting in a mask shortage. Furthermore, their prolonged use causes skin problems related to bacterial overgrowth. To overcome these problems, atmospheric pressure cold plasma was studied as an alternative technology for mask disinfection. Different microorganisms (Pseudomonas aeruginosa, Escherichia coli, Staphylococcus spp.), different gases (nitrogen, argon, and air), plasma power (90-300 W), and treatment times (45 seconds to 5 minutes) were tested. The best atmospheric pressure cold plasma treatment was the one generated by nitrogen gas at 300 W and 1.5 minutes. Testing of breathing and filtering performance and microscopic and visual analysis after one and five plasma treatment cycles, highlighted that these treatments did not affect the morphology or functional capacity of the masks. Considering the above, we strongly believe that atmospheric pressure cold plasma could be an inexpensive, eco-friendly, and sustainable mask disinfection technology enabling their reusability and solving mask shortage.

Carbapenem-resistant Pseudomonas aeruginosa (CRPA) are a global health concern. The antimicrobial resistance, virulence, and molecular typing of 57 CRPA isolated from 43 patients who attended a specific Tunisian hospital from September 2018 to July 2019 were analyzed. All but one were multidrug-resistant CRPA, and 77% were difficult-to-treat-resistant (DTR) isolates. The bla_VIM-2 gene was detected in four strains (6.9%), and among the 36 bla_GES-positive CRPA (62%), the bla_GES-5 gene was the predominant variant (86%). Three strains co-harbored the bla_VIM-2 and bla_GES-45 genes, and seven CRPA carried the bla_SHV-2a gene (14%). OprD alterations, including truncations by insertion sequences, were observed in 18 strains. Regarding the 46 class 1 integron-positive CRPA (81%), the bla_GES-5 gene was located in integron In717, while the bla_GES-29 and bla_GES-45 genes were found in two new integrons (In2122 and In4879), and the bla_VIM-2 gene was found in In1183 and the new integron In2142. Twenty-four PFGE patterns and thirteen sequence types (three new ones) were identified. The predominant serotype O:11 and exoU (81%) were mostly associated with ST235 and the new ST3385 clones. The seven bla_SHV-2a-CRPA from different patients belonged to ST3385 and the same PFGE pattern. The bla_GES-5- and bla_VIM-2 + bla_GES-45-positive CRPA recovered mostly from ICU patients belonged to the high-risk clone ST235. Our results highlight the alarming prevalence of bla_GES-5- and ST235-CRPA, the co-existence of bla_GES-45 and bla_VIM-2, and their location within integrons favoring their dissemination.