This study aimed to evaluate the frequency of isolation of Staphylococcus aureus from different pathological samples processed in the Microbiology Laboratory of the National Institute of Infectious Diseases “Prof. Dr. Matei Balș”, Romania, between 1 January 2017 and 31 December 2022, aiming to establish the ratio of methicillin-resistant to methicillin-susceptible Staphylococcus aureus strains and the antibiotic resistance pattern of isolated microorganisms. The data of isolates originating from routine diagnostic tasks were analyzed retrospectively using laboratory data from the microbiology department. Up to 39.11% of Staphylococcus aureus strains were resistant to oxacillin (MRSA), with 49.97% resistance to erythromycin and 36.06% inducible resistance to clindamycin. Resistance rates to ciprofloxacin, rifampicin, gentamicin, and trimethoprim-sulfamethoxazole were 9.98%, 5.38%, 5.95%, and 0.96%, respectively. There was no resistance to vancomycin. Between 2017 and 2022, the percentage of MRSA strains decreased from 41.71% to 33.63%, sharply increasing to 42.42% in 2021 (the year of the COVID-19 pandemic, when the percentage of strains isolated from lower respiratory tract infections was higher than that of strains isolated from wounds or blood, as in previous years). This study showed a high percentage of MRSA strains (39.11% overall) with a higher proportion of these strains isolated from the blood (42.49%) compared to other clinical specimens.

The ESCPM group (Enterobacter species including Klebsiella aerogenes - formerly Enterobacter aerogenes, Serratia species, Citrobacter freundii complex, Providencia species and Morganella morganii) has not yet been incorporated into systematic surveillance programs. We conducted a multicentre retrospective observational study analysing all ESCPM strains isolated from blood cultures in 27 European hospitals over a three-year period (2020-2022). Diagnostic approach, epidemiology, and antimicrobial susceptibility were investigated. Our study comprised 6,774 ESCPM isolates. MALDI-TOF coupled to mass spectrometry was the predominant technique for bacterial identification. Susceptibility to new β-lactam/β-lactamase inhibitor combinations and confirmation of AmpC overproduction were routinely tested in 33.3% and 29.6% of the centres, respectively. The most prevalent species were E. cloacae complex (44.8%) and S. marcescens (22.7%). Overall, third-generation cephalosporins (3GC), combined third- and fourth-generation cephalosporins (3GC + 4GC) and carbapenems resistance phenotypes were observed in 15.7%, 4.6%, and 9.5% of the isolates, respectively. AmpC overproduction was the most prevalent resistance mechanism detected (15.8%). Among carbapenemase-producers, carbapenemase type was provided in 44.4% of the isolates, VIM- (22.9%) and OXA-48-enzyme (16%) being the most frequently detected. E. cloacae complex, K. aerogenes and Providencia species exhibited the most notable cumulative antimicrobial resistance profiles, with the former displaying 3GC, combined 3GC + 4GC and carbapenems resistance phenotypes in 15.2%, 7.4%, and 12.8% of the isolates, respectively. K. aerogenes showed the highest rate of both 3GC resistant phenotype (29.8%) and AmpC overproduction (32.1%), while Providencia species those of both carbapenems resistance phenotype (42.7%) and carbapenemase production (29.4%). ESCPM isolates exhibiting both 3GC and combined 3GC + 4GC resistance phenotypes displayed high susceptibility to ceftazidime/avibactam (98.2% and 95.7%, respectively) and colistin (90.3% and 90.7%, respectively). Colistin emerged as the most active drug against ESCPM species (except those intrinsically resistant) displaying both carbapenems resistance phenotype (85.8%) and carbapenemase production (97.8%). This study presented a current analysis of ESCPM species epidemiology in Europe, providing insights to inform current antibiotic treatments and guide strategies for antimicrobial stewardship and diagnostics.

Background: The antimicrobial resistance (AMR) of Klebsiella pneumoniae recorded a steep upward trend over the last two decades, among which carbapenem-resistant Klebsiella pneumoniae (CRKP) is one of the most concerning strains considering the development and spread of AMR. The aim of this study was to analyze the evolution of AMR for Klebsiella pneumoniae and to describe the risk factors of AMR for Klebsiella pneumoniae, including the COVID-19 pandemic. Methods: We conducted a retrospective study on Klebsiella pneumoniae non-duplicative isolates collected from patients admitted to a tertiary hospital in Bucharest, Romania, from January 2019 to December 2021. We evaluated AMR changes by comparing resistance between 2019 and the mean of 2020–2021. Results: The rates of AMR increased for third-generation cephalosporins, carbapenems, aminoglycosides, fluoroquinolones, and colistin and decreased for trimethoprim/sulfamethoxazole (TMP/SMX), 45.7% in 2019 vs. 28.3% in 2021. A longer length of hospital stay (ꭓ² = 49.68, p < 0.01); recent antibiotic consumption, RR = 1.38, 95% CI [1.21, 1.57]; and recent contact with hospital settings, RR = 1.54, 95% CI [1.32, 1.8] were risk factors for multidrug-resistant (MDR) Klebsiella pneumoniae. Conclusions: The AMR of Klebsiella pneumoniae increased during 2020–2021 for most of the potential active antibiotics; only TMP/SMX resistance decreased, and it may represent a treatment option for CRKP or MDR Klebsiella pneumoniae infections. Decreasing the excessive use of antibiotics and the implementation of prevention and control measures in healthcare settings are mandatory for avoiding further increases in the AMR rate of Klebsiella pneumoniae.

Background: Romania is one of the European countries reporting very high antimicrobial resistance (AMR) rates and consumption of antimicrobials. We aimed to characterize the AMR profiles and clonality of 304 multi-drug resistant (MDR) Acinetobacter baumannii (Ab) and Pseudomonas aeruginosa (Pa) strains isolated during two consecutive years (2018 and 2019) from hospital settings, hospital collecting sewage tanks and the receiving wastewater treatment plants (WWTPs) located in the main geographical regions of Romania. Methods: The strains were isolated on chromogenic media and identified by MALDI-TOF-MS. Antibiotic susceptibility testing and confirmation of ESBL- and CP- producing phenotypes and genotypes were performed. The genetic characterization also included horizontal gene transfer experiments, whole-genome sequencing (WGS), assembling, annotation and characterization. Results: Both clinical and aquatic isolates exhibited high MDR rates, especially the Ab strains isolated from nosocomial infections and hospital effluents. The phenotypic resistance profiles and MDR rates have largely varied by sampling point and geographic location. The highest MDR rates in the aquatic isolates were recorded in Galați WWTP, followed by Bucharest. The Ab strains harbored mostly bla_OXA-23, bla_OXA-24, bla_SHV, bla_TEM and bla_GES, while Pa strains bla_IMP, bla_VIM, bla_NDM, bla_VEB, bla_GES and bla_TEM, with high variations depending on the geographical zone and the sampling point. The WGS analysis revealed the presence of antibiotic resistance genes (ARGs) to other antibiotic classes, such as aminoglycosides, tetracyclines, sulphonamides, fosfomycin, phenicols, trimethoprim-sulfamethoxazole as well as class 1 integrons. The molecular analyses highlighted: (i) The presence of epidemic clones such as ST2 for Ab and ST233 and ST357 for Pa; (ii) The relatedness between clinical and hospital wastewater strains and (iii) The possible dissemination of clinical Ab belonging to ST2 (also proved in the conjugation assays for bla_OXA-23 or bla_OXA-72 genes), ST79 and ST492 and of Pa strains belonging to ST357, ST640 and ST621 in the wastewaters. Conclusion: Our study reveals the presence of CP-producing Ab and Pa in all sampling points and the clonal dissemination of clinical Ab ST2 strains in the wastewaters. The prevalent clones were correlated with the presence of class 1 integrons, suggesting that these isolates could be a significant reservoir of ARGs, being able to persist in the environment.

Background: New Delhi metallo-beta-lactamase (NDM)-producing Providencia stuartii has been reported from European Union/European Economic Area (EU/EEA) countries with increasing frequency. During 2018 to 2022, 355 cases of NDM-producing P. stuartii were detected in seven hospitals reporting on NDM-production in Enterobacterales in Romania. Aim: Our aim was to determine the extent of spread of NDM-producing P. stuartii in hospitals in Romania. Methods: We analysed whole genome sequences and epidemiological data of 74 P. stuartii isolates collected in six hospitals from December 2021 to September 2023. Results: We identified four multi-hospital clusters including isolates detected over more than a year, indicating sustained spread of bla _NDM-1-carrying P. stuartii within the healthcare system. These clusters consisted of isolates from up to four hospitals and three regions. Three multi-hospital clusters were caused by a specific multidrug-resistant P. stuartii sequence type 46 lineage carrying bla _NDM-1 and a large set of additional resistance markers. Investigation in an international context showed that this lineage had already been detected in nine countries (Bulgaria, France, Germany, Ireland, the Netherlands, Romania, Switzerland, United Kingdom, United States) since 2015. Conclusion: Our results alert about the risk of carbapenem-resistant P. stuartii transmission in healthcare settings. Enhanced infection prevention and control measures should be instituted as soon as cases are detected in healthcare facilities. National surveillance systems in EU/EEA countries should, in addition to carbapenem-resistant and/or carbapenemase-producing Klebsiella pneumoniae and Escherichia coli, consider reporting carbapenem-resistant and/or carbapenemase-producing P. stuartii and other Enterobacterales where relevant.

Background In aesthetic breast augmentation, especially by the transareolar approach, there is increasing concern regarding the occurrence of capsular contracture and its potential correlation with intraoperative implant contamination from putative endogenous breast flora of the nipple and lactiferous ducts. However, detectable bacteria cannot be considered synonymous with established resident microflora. Objectives The authors sought to elucidate the existence of endogenous breast flora and assess the microbiologic safety of transareolar breast augmentation. Methods In this prospective study (BREAST-MF), the authors collected microbiologic samples from the breast skin, ductal tissue, and parenchyma of 39 consecutive female patients who underwent breast procedures in a plastic surgery clinic. Swabs collected pre-, intra-, and postoperatively were processed for bacterial and fungal growth. Positive cultures underwent identification through VITEK and MALDI-TOF, as well as antimicrobial susceptibility testing. Results Staphylococcus species accounted for 95 of 106 (89.6%) positive results from native breast skin, 15 of 18 (83.3%) positive results from decontaminated breast skin, and 4 of 4 (100%) positive results from the breast parenchyma. Methicillin resistance was present in 26.4% of S. epidermidis, 25.3% of S. hominis, and 71.4% of S. haemolyticus strains. Conclusions During transareolar breast augmentation, in the nipple-areola region it is more likely to find bacteria populating the skin, rather than endogenous breast flora, as previously considered. Appropriate preoperative decontamination is essential for minimizing the risk of postoperative infections. Level of Evidence 3 Risk