High Prevalence of GES-5 Variant and Co-Expression of VIM-2 and GES-45 among Clinical Pseudomonas aeruginosa Strains in Tunisia.

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 blaVIM-2 gene was detected in four strains (6.9%), and among the 36 blaGES-positive CRPA (62%), the blaGES-5 gene was the predominant variant (86%). Three strains co-harbored the blaVIM-2 and blaGES-45 genes, and seven CRPA carried the blaSHV-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 blaGES-5 gene was located in integron In717, while the blaGES-29 and blaGES-45 genes were found in two new integrons (In2122 and In4879), and the blaVIM-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 blaSHV-2a-CRPA from different patients belonged to ST3385 and the same PFGE pattern. The blaGES-5- and blaVIM-2 + blaGES-45-positive CRPA recovered mostly from ICU patients belonged to the high-risk clone ST235. Our results highlight the alarming prevalence of blaGES-5- and ST235-CRPA, the co-existence of blaGES-45 and blaVIM-2, and their location within integrons favoring their dissemination.

Phenotypic Characteristics and Clonal Relationships of Stenotrophomonas maltophilia Isolates in Hospitalized Adults from a Private Center in Lima, Peru.

Stenotrophomonas maltophilia is an opportunistic pathogen, often associated with nosocomial infections. Ten S. maltophilia were isolated from clinical samples during the period January 2021 and June 2022. Eight (80%) patients had cancer as a background disease and 2 patients had coronavirus disease 2019. A fatal outcome was recorded in 4 cases (40% of patients). All the isolates were susceptible to minocycline and levofloxacin. Trimethoprim/sulfamethoxazole and ceftazidime resistance rates were 20% and 40% respectively. Eight different patterns were observed by Pulsed-Field Gel Electrophoresis, only two isolates being clonally identical. The isolation of S. maltophilia in clinical settings requires the implementation of infection prevention measures.

Mask disinfection using atmospheric pressure cold plasma.

OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Development and characterization of anti-biofilm coatings applied by Non-Equilibrium Atmospheric Plasma on stainless steel.

Biofilm-mediated microbial persistence of pathogenic and spoilage bacteria is a serious problem in food industries. Due to the difficulty of removing mature biofilms, great efforts are being made to find new strategies to prevent bacterial adherence to surfaces, the first step for biofilm development. In this study, coatings of (3-aminopropyl)triethoxysilane (APTES), tetraethyl orthosilicate (TEOS) and acrylic acid (AA) were applied by Non-Equilibrium Atmospheric Plasma on stainless steel (SS) AISI 316, the SS most commonly used in food industry equipment. Their anti-biofilm activity was assessed against Listeria monocytogenes CECT911 and Escherichia coli CECT515 after incubation at 37 °C. The best results were obtained for L. monocytogenes, with coatings consisting of a base coating of APTES and a functional coating of TEOS (AP10 + TE6) or AA (AP10 + AA6) that reduced biofilm production by 45% and 74%, respectively, when compared with the uncoated SS. These coatings were further characterized, together with a variation of the best one that replaced the acrylic acid with succinic acid (AP10 + SA6). Their anti-biofilm activity was assessed under different incubation conditions, including two strains of L. monocytogenes isolated from processing environments of a meat industry. The coating AP10 + AA6 reduced the biofilm formation by 90% after incubation at 12 °C, a temperature more representative of those commonly found in food processing environments. The morphological and physico-chemical characterization of the selected coatings showed that the coating with the highest anti-biofilm activity (i.e., AP10 + AA6) had lower surface roughness and higher hydrophilicity. This suggests that the formation of a hydration layer prevents the adherence of L. monocytogenes, an effect that seems to be enhanced by low temperature conditions, when the wettability of the strains is increased.

Streptococcus dysgalactiae subsp. equisimilis from invasive and non-invasive infections in Spain: combining epidemiology, molecular characterization, and genetic diversity.

The purpose of this study was to characterize the antibiotic resistance, virulence, and genetic diversity among invasive and non-invasive Streptococcus dysgalactiae subsp. equisimilis (SDSE) isolates. SDSE were isolated from clinical samples of outpatients and inpatients cares in La Rioja region (Spain) during 2012-2015. The analyses performed were susceptibility testing by disc diffusion, resistance and virulence genes by PCR, emm typing by PCR and sequencing, and other molecular typing by SmaI-PFGE and MLST. Forty-two SDSE isolates were recovered (64.3% non-invasive, 35.7% invasive) that were grouped in 31 PFGE patterns, 17 ST, and 14 emm types, being stC1400, stG6792, and stG62647 the most frequent, and stC74a and stC5345 exclusive in invasive SDSE. Twenty-one SDSE were resistant to at least one antibiotic. The erm(TR) and erm(B) genes were linked with resistance to macrolides; tet(M) and tet(T) to tetracycline; dfrF to trimethoprim; ant(6)-Ia and aph(3')-IIIa to aminoglycosides; and the substitutions Asp80Ala in GyrA and Ser79Phe in ParC with resistance to levofloxacin. The sagA, slo, scpA, and ska virulence genes were amplified in 93% SDSE. Streptococcal superantigenic speGdys gene was identified in 80% of invasive and 63% of non-invasive SDSE and correlated with certain emm types (e.g., stG62647 or stG6792). SDSE invasive infections were most frequent in elderly patients, and half of our SDSE were resistant to at least one antibiotic tested. This work is the first detection of tet(T), dfrF, and new substitution in GyrA protein in SDSE. A high diversity of circulating genetic lineages was found among our SDSE.

Antibiofilm coatings through atmospheric pressure plasma for 3D printed surgical instruments.

Recently, medical applications for 3D printing are expanding rapidly and are expected to revolutionize health care, specifically, manufacturing surgical guides and protective face mask against coronavirus (COVID-19). These instruments come in contact with the human tissues, being necessary 3D printed materials free of pathogenic microbes or other contaminants. Therefore, they must be sterilized to avoid that bacteria can attach to the surface and produce biofilm. With the aim of avoiding bacterial biofilm formation and minimize the health risks, acrylic acid (AcAc) coatings applied by plasma-polymerization have been deposited on 3D printed polylactic acid (PLA) Petri dishes. Six antimicrobial-resistant clinical and two susceptible control strains of Pseudomonas aeruginosa and Staphylococcus aureus species were analyzed. AcAc coatings provide the surface with greater hydrophilicity and, consequently, the formation of a hydration layer, whose thickness is related to the surface roughness. This hydration layer could explain the reduction of bacterial attachment and, consequently, the biofilm formation. Antibiofilm coatings are more successful against P. aeruginosa strains than against S. aureus ones; due to some coatings presents a smaller topography scale than the P. aeruginosa length, reducting the contact area between the bacteria and the coating, and causing a potential rupture of the cellular membrane. AcAc coatings with less number of plasma passes were more effective, and showed up to a 50% relative biofilm reduction (in six of the eight strains studied) compared with the untreated plates.

Antimicrobial resistance and virulence of Pseudomonas spp. among healthy animals: concern about exolysin ExlA detection.

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.