Background: This study provides a comprehensive analysis of Acinetobacter baumannii in aquatic environments and fish microbiota by integrating culture-dependent methods, 16S metagenomics, and antibiotic resistance profiling. Methods: A total of 83 A. baumannii isolates were recovered using culture-dependent methods from intra-hospital infections (IHI) and wastewater (WW) and surface water (SW) samples from two southern Romanian cities in August 2022. The antibiotic susceptibility was screened using disc diffusion, microdilution, PCR, and Whole Genome Sequencing assays. Results: The highest microbial load in the analyzed samples was found in Glina, Bucharest, for both WW and SW samples across all investigated phenotypes. For Bucharest isolates, the resistance levels corresponded to fluoroquinolones > aminoglycosides > β-lactam antibiotics. In contrast, A. baumannii from upstream SW samples in Târgoviște showed the highest resistance to aminoglycosides. The bla_OXA-23 gene was frequently detected in IHI, WW, and SW isolates in Bucharest, but was absent in Târgoviște. Molecular phylogeny revealed the presence of ST10 in Târgoviște isolates and ST2 in Bucharest isolates, while other minor STs were not specifically correlated with a sampling point. Using 16S rRNA sequencing, significant differences in microbial populations between the two locations was identified. The low abundance of Alphaproteobacteria and Actinobacteria in both locations suggests environmental pressures or contamination events. Conclusions: These findings indicate significant fecal contamination and potential public health risks, emphasizing the need for improved water quality monitoring and management.

Nosocomial fungal infections caused by Candida auris pose a threat to public health due to their increased resistance to common antifungal drugs. Four thiourea derivatives of 2-thiophenecarboxylic acid were evaluated for their antifungal and antioxidant activity. The antifungal activity of the compounds was tested against strains of C. auris isolated from a hospital in Romania. With a notable inhibitory effect on C. auris biofilm growth and microbial adherence, the ortho-methylated derivative (SB2) showed the highest antifungal activity. Furthermore, emphasizing the impact of structural factors on the electron-donating capacity of these compounds, antioxidant activity assays (DPPH, FRAP, TEAC and CUPRAC) identified the SB2 compound as having the highest antihemolytic and antioxidant effects. The low cytotoxicity validated by hemocompatibility assays makes these compounds options for antifungal treatment. The results show that antifungal and antioxidant action is greatly influenced by structural modifications, especially the position of the methyl group on the aromatic ring. The possible clinical uses of these molecules as drugs for the treatment of multidrug-resistant C. auris infections needs further investigation.

Background/Objectives: This study aims to characterize antibiotic resistance (AR) and virulence markers in Salmonella spp. isolated from Romanian outpatients’ stool samples. Methods: In 2019, community-acquired Salmonella strains were collected and identified using MALDI-TOF mass spectrometry, antibiotic susceptibility profiles have been determined with the MicroScan system, and soluble virulence factors were evaluated using specific culture media, while biofilm formation was quantified in 96-well plates. Molecular analysis targeted resistance genes for β-lactams (e.g., bla_TEM and bla_SHV); tetracyclines (e.g., tet(A)); sulphonamides; and quinolones, as well as virulence genes (e.g., invA, spvC, pldA, and held). Whole-genome sequencing (WGS) was performed on 19 selected isolates. A silver nanoparticles (AgNPsol) alternative to conventional antibiotics was tested for effectiveness against multidrug-resistant (MDR) isolates. Results: From the total of 309 Salmonella isolates (65.05% from children under 4 years of age) belonging to four subtypes and four serovars, 27.86% showed resistance to at least one antibiotic, most frequently to tetracycline, ampicillin, and piperacillin. The strains frequently expressed haemolysin (67%), aesculinase (65%), and gelatinase (62%). Resistance to trimethoprim-sulfamethoxazole was encoded by the sul1 gene in 44.83% of the strains and to tetracyclines by the tet(A) gene (59.52%). The ESBL genes bla_TEM, bla_SHV, and bla_CTX-M were detected by PCR in 16.18%, 2.91%, and 0.65% of the strains, respectively. Additionally, 98.63% of the strains carried the invA marker, with notable positive associations between bla_SHV, qnrB, and sul1 with spvC. Conclusions: The present findings revealed significant patterns in Salmonella isolates, subtypes, serovars, AR, and virulence, emphasising the need for continuous surveillance of Salmonella infections. Additionally, the potential of AgNPs as an alternative treatment option was demonstrated, particularly for paediatric S. enterica infections.