Genomic and epidemiological insight of an outbreak of carbapenemase-producing Enterobacterales in a Portuguese hospital with the emergence of the new KPC-124.

BACKGROUND: Carbapenemase-producing Enterobacterales (CPE) is an increasing problem in healthcare settings. This study aimed to identify the source of a CPE outbreak that occurred in 2022, in a tertiary hospital in the North of Portugal, to identify exposed patients, and to assess the risk of becoming CPE-positive following hospital admission. METHODS: A multi-disciplinary investigation was conducted including descriptive, analytical, and molecular epidemiology, environmental screening, and assessment of infection control measures. Clinical and environmental isolates were analyzed using whole-genome sequencing and phylogenetic analysis. Additionally, a prospective observational cohort study was conducted to further investigate the risk factors associated with the emergence of new cases in cohorts of CPE-negative admitted patients. RESULTS: We observed the presence of multispecies KPC-, IMP-, and/or NDM-producing isolates. Genetically indistinguishable clinical and environmental isolates were found on the same room/ward. The ST45 KPC-3-producing Klebsiella pneumoniae clone was the responsible for the outbreak. During patients' treatment, we detected the emergence of resistance to ceftazidime-avibactam, associated with mutations in the blaKPC-3 gene (blaKPC-46, blaKPC-66 and blaKPC-124, the last variant never previously reported), suggesting a vertical evolutionary trajectory. Patients aged ≥ 75 years, hygiene/feeding-care dependent, and/or subjected to secretion aspiration were risk factors for CPE colonization after hospital admission. Additionally, cases with previous admission to the emergency department suggest that CPE dissemination may occur not only during hospitalization but also in the emergency department. CONCLUSION: Overall, the study highlights that selection pressure with antibiotics, like ceftazidime-avibactam, is a contributing factor to the emergence of new ß-lactamase variants and antibiotic resistance. It also shows that the hospital environment can be a significant source of CPE transmission, and that routine use of infection control measures and real-time molecular epidemiology investigations are essential to ensure the long-term termination of CPE outbreaks and prevent future resurgences.

Airborne spores' dissemination of a swine associated Clostridioides difficile clone.

This study supports the airborne dissemination of Clostridioides difficile spores. Of the sieve impaction samples collected at a swine production unit, 66.7% were positive and all belonged to the predominantly established clone. Spores' density varied according to the characteristics of the animal population, suggesting the possibility of airborne transmission.

Occurrence of polycyclic aromatic hydrocarbons, microplastics and biofilms in Alqueva surface water at touristic spots.

Freshwater pollution is a huge concern. A study aiming to evaluate physico-chemical characteristics, microbiota, occurrence of two groups of persistent environmental pollutants with similar chemical properties (polycyclic aromatic hydrocarbons- PAHs and microplastics - MPs) in Alqueva's surface water was performed during 2021. Water samples were collected at three spots related to touristic activities (two beaches and one marina) during the Winter, Spring, Summer and Autumn seasons. In addition, the presence of biofilms on plastic and natural materials (stone, wood/ vegetal materials) were assessed and compared. Water quality based on physicochemical parameters was acceptable with a low eutrophication level. PAHs concentration levels were lower than the standard limits established for surface waters by international organizations. However, carcinogenic compounds were detected in two sampling locations, which can pose a problem for aquatic ecosystems. PAHs profiles showed significant differences when comparing the dry seasons with the rainy seasons, with a higher number of different compounds detected in Spring. Low molecular weigh compounds, usually associated with the atmospheric deposition and petroleum contamination, were more prevalent. MPs were detected in all samples except one during the Winter season. The polymers detected were poly(methyl-2-methylpropenoate), polystyrene, polyethylene terephthalate, polyamide, polypropylene, styrene butadiene, polyvinyl chloride and low /high density polyethylene with the last being the most frequent. Biofilms were more often detected on plastics than on natural materials. In addition, biofilms detected on plastics were more complex with higher microbial diversity (e.g., bacteria, fungi/yeast and phytoplancton organisms) and richer in extrapolymeric material. Based on morphological analysis a good agreement between microbiota and microorganism present in the biofilms was found. Among microbiota were identified microorganisms previously linked to plastic and PAHs detoxification suggesting the need for further studies to evaluate the viability of using biofilms as part of a green bioremediation strategy to mitigate water pollution.

The impact of indoor air quality on respiratory health of older people living in nursing homes: spirometric and exhaled breath condensate assessments.

In the Portuguese Geriatric Study of the Health Effects of Indoor Air Quality in Senior Nursing Homes, we aimed to evaluate the impact of indoor air contaminants on the respiratory symptoms and biomarkers in a sample of elderly living in nursing homes. A total of 269 elderly answered a health questionnaire, performed a spirometry and 150 out of these collected an exhaled breath condensate sample for pH and nitrites analysis. The study included the evaluation of indoor chemical and microbiological contaminants. The median age of the participants was 84 (78-87) years and 70.6% were women. The spirometric data indicated the presence of airway obstruction in 14.5% of the sample. Median concentrations of air pollutants did not exceed the existing standards, although increased peak values were observed. In the multivariable analysis, each increment of 100 µg/m3 of total volatile organic compounds was associated with the odds of respiratory infection in the previous three months ( OR̂ =1.05; 95% CI: 1.00-1.09). PM2.5 concentrations were inversely associated with pH values ( ß̂ = -0.04, 95%: -0.06 to -0.01, for each increment of 10 µg/m3). Additionally, a direct and an inverse association were found between total bacteria and FEV1/FVC and FVC, respectively.

Environmental and ventilation assessment in Child Day Care Centers in Porto: the ENVIRH Project.

Children attending day care centers (CDCC) have been reported to be more prone to infectious diseases when compared with those cared for at home, and are exposed to conditions that may increase the risk of allergies and asthma. Several studies revealed that consequences of poor ventilation conditions include high levels of carbon dioxide (CO2) and many other indoor pollutants commonly detected in schools. Nine child day care centers were selected randomly to participate in this study. Fifty-two classrooms were assessed for chemical, biological, physical, and allergen parameters in spring and winter seasons in these nine CDCC located in Porto, Portugal. Outdoor measurements were also conducted for comparison. Our results indicated that (i) particulate matter (PM10) median levels were above the national reference levels, both by classroom type and by season; (ii) TVOC kindergarten peak values may raise some concern; (iii) CO2 was present at high median and maximum levels during spring and winter assessment in both nurseries and kindergartens classrooms; (iv) total bacteria concentrations were 57- and 52-fold higher in the nursery and kindergarten than outdoors, respectively, for the spring season; (v) winter and spring median predicted mean vote (PMV) indices were between "neutral" (0) and "slightly cool" (≤ -1) in the thermal sensation scale for comfort situations (-2 to 2) for both types of classrooms; (vi) there were significant differences for both PMV and predicted percentage of dissatisfied (PPD) indices by season; and (vii) CO2, total bacteria, and gram-negative bacteria were associated with low airflow rates. These data will help to evaluate the effectiveness of current building operation practices in child day care centers regarding indoor air quality and respiratory health.

Reduced graphene oxide: firm support for catalytically active palladium nanoparticles and game changer in selective hydrogenation reactions.

Simultaneous decomposition and reduction of a Pd(2+) complex in the presence of graphene oxide (GO) lead to the formation of Pd(0)-nanoparticles (Pd-NPs) with average sizes of 4 nm firmly anchored on reduced graphene oxide (RGO) sheets. The Pd-NP/RGO hybrids exhibited remarkable catalytic activity and selectivity in mild hydrogenation reactions where the acidic properties of RGO play an active role and may act as an important game-changer.