Introduction: During the early stages of the COVID-19 pandemic—when many health services were inaccessible—the role of community pharmacists became particularly prominent, playing key roles in pharmaceutical care and patient education. This study aimed to explore the perceptions and attitudes of community pharmacy staff regarding the conditions of medication supply and patterns of self-medication, with a focus on antimicrobials, during the first year of the COVID-19 pandemic.

Methods: A qualitative, descriptive study was conducted using semi-structured, focus-group interviews with community pharmacy staff in the Southern Great Plain of Hungary. Data collection was carried out between 15/05/2020 and 15/02/2021, using a non-probability sampling approach. Interviews were conducted online via the Zoom™ videoconference platform, transcribed manually, and analysed via thematic analysis through an inductive approach. A total of N=16 pharmacists participated across four focus groups (3–5 participants per group). Recruitment continued until saturation was achieved, i.e. when no new relevant information emerged. The study followed the Consolidated criteria for Reporting Qualitative Research (COREQ) guidelines.

Results: Of the participants, 11/16 were women, and their median age was 35 years (range: 26–57); 6/16 had board certifications in pharmacy management. The following major themes were identified: (i) the role of the National eHealth Infrastructure (EESzT) and e-prescriptions during the pandemic; (ii) the emergence and monitoring of COVID-19 “miracle cures” (e.g., hydroxychloroquine, ivermectin, azelastine) and medicines “to be avoided” (e.g., ibuprofen, ACE inhibitors); and (iii) the legitimization of inappropriate antibiotic use through "defensive medicine”.

Conclusions: Rapid dissemination of health-related information—regardless of its basis in evidence—shaped public interest in the perceived efficacy and availability of certain medicines; the trend of "information noise” generated by the media was clearly reflected in community pharmacies. Patterns of antibiotic self-medication have also shifted, as many patients obtained antimicrobials through physician prescriptions issued for “prophylactic” purposes during the pandemic.

Background and Aims
In the delicate realm of neonatal care, peripheral venous catheters (PVCs) are indispensable but also serve as potential gateways for catheter-associated infections (CAIs). These infections, often caused by Staphylococcus aureus, including methicillin-resistant strains (MRSA), contribute significantly to the growing challenge of bacterial resistance in hospital settings. This study aimed to assess catheter colonization, characterize resistance profiles—especially methicillin resistance—and identify factors linked to infection risk. Particular attention was given to Staphylococcus spp. and their resistance mechanisms.

Methods
We collected 403 catheter tip samples from 325 neonates hospitalized in NICUs. Bacterial colonization was evaluated using the quantitative Cleri–Brun–Buisson method. Emphasis was placed on S. aureus. Methicillin resistance was confirmed through PCR detection of the mecA gene. We conducted a genetic diversity analysis of mecA sequences to explore clonal variation. Additionally, the D-test was used to detect inducible clindamycin resistance, offering insight into phenotypic resistance expression.

Results
Colonization by MRSA and methicillin-resistant CoNS was identified in a subset of samples. Prolonged catheter dwell time and inconsistent aseptic technique were associated with higher colonization rates. The mecA gene was detected in several isolates, displaying notable genetic diversity indicative of multiple circulating clones. The D-test revealed inducible clindamycin resistance in various strains, underscoring the relevance of phenotypic testing.

Conclusion
Our findings highlight the need to reinforce infection prevention protocols and closely monitor bacterial resistance in NICUs. The diversity of mecA-positive strains and the detection of inducible resistance patterns underscore the importance of combining molecular and phenotypic methods. Improved catheter management and targeted surveillance can significantly reduce CAIs and enhance the protection of vulnerable neonatal patients.

Aim and scope

In compliance with the One Health concept, it is important to look at the use of antibiotics as a whole, i.e. the use of both human and veterinary antibiotics together. According to the AWaRe classification of antibiotics for human use and the AMEG categorisation of antibiotics for use in animals, antibiotics are divided into different categories based on their resistance risk and medical importance. The same ingredient may be classified in the lowest risk category for humans and the risk for public health in veterinary medicine is estimated higher. The aim is comparing the selection of antimicrobial ingredients in treatment of humans by AWaRe classification and in treatment of companion animals by AMEG categorisation. This analysis includes data of human and veterinary antibiotics in pharmaceutical form as tablets.

Results and conclusions

In 2020–2024, 36 different antimicrobial ingredients were used in human treatment and 28 in companion animal treatment, 7 of them for both: Amoxicillin, Amoxicillin/clavulanic acid, Cefadroxil, Cefalexin, Clindamycin, Doxycycline, Metronidazole. Both financially and quantitatively, the most sold antibiotic for humans and companion animals was Amoxicillin/clavulanic acid, classified for humans by AWaRe classification as “Access” (green, first-line) and for animals by AMEG classification as “Caution” (yellow, limited use) category. Amoxicillin/clavulanic acid accounted for 23% of the total cost of human antibiotics and for 64% of the total cost of antibiotics used in treatment of companion animals. The overall sales of AWaRe category “Access” antibiotics accounted for 78% of total human antibiotics, and the sales of AMEG category “Caution” antibiotics accounted for 90% of all antibiotics used for companion animals, in Estonia.

AWaRe and AMEG classifications are intended as tools for optimizing and monitoring antibiotic use, also for monitoring the impact of stewardship policies implemented to manage antimicrobial resistance.

Enterobacterale strains like E. coli, Klebsiella pneumoniae, and Enterobacter spp. pose a serious epidemiological threat due to their potential as pathogenic organisms. Furthermore, bacteria belonging to the order Enterobacterales frequently exhibit resistance to carbapenem antibiotics and are capable of producing extended-spectrum β-lactamases (ESBLs). Consequently, they have been designated as critical priority pathogens by the World Health Organization (WHO, 2024).

Therefore, this study aimed to determine the impact of treated wastewater discharge on bacterial biodiversity within the order Enterobacterales and the presence of carbapenem resistance genes (CRGs) in river water samples at the catchment level.

River water was sampled at eight points from the upper Pilica River to its confluence with the Vistula (Poland). A total of 32 samples in four seasons were collected. Metagenomic DNA was extracted from the samples, and single-read Illumina sequencing was carried out. The number of CRGs in the samples was established through digital PCR. Physico-chemical parameters were studied in accordance with standard methods.

In the current study, the prevalence of Enterobacterales was lower than that of the other identified taxa. Enterobacterales accounted for about 1% of all amplicon sequence variants (ASVs), including Escherichia-Shigella (65.97% of Enterobacterales ASVs), Yersinia spp. (10.63%), Klebsiella spp. (9.69%), Enterobacter spp. (7.17%), Serratia spp. (4.21%) and others. All studied CRGs were present in the analysed samples. The most prevalent genes in river samples were bla_VIM and bla_OXA, while other studied genes (bla_IMP, bla_NDM, and bla_KPC) occurred at lower concentrations. The research confirmed a relationship between selected Enterobacterales ASVs and CRG concentrations in the analysed samples. Moreover, we observed a statistically significant relationship between the studied micropollutants and temperature, as well as BOD5, COD, total phosphorus and nitrogen.

Concentrations of all studied micropollutants, including CRGs, Enterobacterales ASVs, and physico-chemical parameters, were sensitive indicators of anthropogenic influence in river water.

The overuse of antimicrobials in humans and animals is driving the emergence of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs), which spread between the One Health compartments: humans, animals, and the environment (water, air, soil). The extent to which surface waters play a role in the dissemination of ARB/ARGs is poorly understood, but recreational use and drinking water may result in human exposure. This is the first large scale survey of the prevalence of cefotaxime-resistant E. coli (CTX-EC) and ARGs in drinking water sources and final water in Scotland.

Drinking water sources with different catchments were selected to determine the effect of faecal pollution sources (human, livestock, agriculture, and wildlife) on CTX-EC and ARGs. Samples from 29 drinking water sources and three final waters (post-treatment) were analysed. Total E. coli and CTX-EC were enumerated and HTqPCR was used to quantify 56 ARGs.

CTX-EC were detected in seven drinking water sources (24%). In the majority (95%) of samples positive for CTX-EC, the proportion of CTX-EC to total E. coli was low (<2%). Logistic regression found significant associations between CTX-EC presence and two catchment variables: septic tanks and pastures. ARGs were present in 90% of drinking water sources and 44 ARGs from eight antibiotic classes were quantified; however, mean relative abundances of ARGs were low (3E-06 to 0.006 ARG copies per 16S rRNA gene). In final waters, all ARGs were below the limit of detection, and no E. coli or CTX-EC were recovered.

This study found that the overall AMR load in Scottish drinking water sources is low. CTX-EC concentrations were low, and their prevalence was around half of that found in coastal waters in Scotland. ARGs were widely detected in drinking water sources, but their relative abundances were low, and drinking water treatment processes effectively removed them.

Subclinical mastitis caused by Staphylococcus aureus remains a major challenge due to persistence, recurrence and antimicrobial use. Bacterial adaptation to mammary gland conditions and intracellular survival contribute to therapeutic failure. Rifaximin α (RIFα), a semisynthetic rifamycin widely used for intramammary therapy, exhibits strong antibacterial activity; however, an integrated pharmacodynamic evaluation considering both extracellular and intracellular compartments is limited.

In vitro pharmacodynamic studies were performed to characterize the extracellular and intracellular activity of RIFα against bovine S. aureus. Extracellular activity was assessed using minimum inhibitory concentration determination, post-antibiotic effect, and time–kill curves under mammary-simulated conditions, including different pH values (7.4, 6.5 and 5.0) and media supplemented with bovine serum or milk. Antibacterial activity was quantified using the antibacterial effect index (E). Intracellular activity was evaluated in polymorphonuclear cells (PMNs) isolated from bovine blood. PMNs were infected with S. aureus ATCC 29213, extracellular bacteria were eliminated, and infected cells were exposed to RIFα at 3×MIC and 10×MIC. Intracellular viable bacteria were quantified by colony-forming unit enumeration following cell lysis.

RIFα demonstrated potent extracellular activity, achieving stable bactericidal effects (E ≤ –3) at 4–8×MIC across pH conditions and in the presence of milk or serum, confirming high antibacterial efficacy under mammary-relevant conditions. In contrast, intracellular exposure resulted in only partial reductions in intracellular S. aureus, with decreases of approximately 1.5–1.7 log₁₀ CFU after 8 h, without reaching bactericidal thresholds.

From a PK/PD standpoint, the contrast between potent extracellular bactericidal activity and limited intracellular efficacy underscores the importance of early exposure-driven antimicrobial action. These findings emphasize the value of dynamic pharmacodynamic approaches that integrate both extracellular and intracellular compartments to better inform rational dosing strategies against persistent Staphylococcus aureus infections.

Laguna de Bay, the largest lake in the Philippines, is fed by 21 tributary river systems and serves as a multi-use water resource for economic and recreational purposes. Backyard farms are also prevalent in the communities around the lake. Unregulated use of antibiotics in these farms and local communities contributes to the emergence of antimicrobial-resistant (AMR) bacteria, posing a significant threat to public health and the environment. While there are existing studies on AMR in the Philippines, research on the prevalence of bacteria harboring antimicrobial-resistance genes (ARGs) in environmental waters and animals remains limited. Therefore, this study aimed to determine the antibiotic resistance profiles of Escherichia coli isolated from selected stations of Laguna Lake, 11 surrounding tributary rivers, and backyard farms near the lake. Environmental water and fecal samples were collected from these sites, and a total of 5,051 E. coli were isolated and genotypically screened for the presence of ARGs: β-lactamase (blaCTX-M, blaSHV, blaTEM, and blaOXA-1), carbapenemase (blaNDM), sulfonamide (sul1 and sul2), and tetracycline (tetA). Among these, tetA was the most frequently detected gene in both environmental water (31%) and animal hosts (41%), followed by blaTEM (27%; 32%), sul2 (16%; 19%), sul1 (9%; 5%), blaSHV (4%; 2%), and blaCTX-M (2%; 2%). blaOXA-1 and blaNDM had lower detection frequencies (>1%) in environmental water and were absent in any animal hosts. Among isolates harboring at least two ARGs, the blaTEM/tetA (n=405) combination was the most frequently detected in water samples. In animals, tetA/blaTEM gene combination was most frequent in poultry and swine, while tetA-blaTEM-sul2 combination dominated in ducks. Overall, the results confirm the presence of ARGs in E. coli isolates from Laguna de Bay, its tributaries, and surrounding farms, which can be used as a reference to monitor environmental contamination and mitigate the spread of antimicrobial resistance.

The burden of increased antimicrobial resistance in microorganisms has complicated disease management and healthcare systems. Mechanisms like enzymatic modification of antibiotics, biofilm formation and genetic mutations enable pathogens to evade antibiotic treatments. In this view, alternatives for antimicrobial agents from natural sources are a subject of concern and should be explored immediately. Bacteriocins are antimicrobial peptides with targeted activity against specific bacteria. They are effective at low concentrations (Soltani et al., 2022) and demonstrate both narrow- and broad-spectrum activity. Bacteriocins selectively target and inhibit pathogenic bacteria, minimizing the development of resistance and preserving beneficial microbiota through quorum sensing. Their molecular weight ranges from <5 to 80 kDa with a smaller number of amino acids according to RiPP nomenclature of bacteriocin classification. Our study has emphasized isolation of novel peptides from the species of Levilactobacillus brevis through microbiological techniques and their purification using Prep-HPLC and characterization by biomolecular techniques—High-Performance Thin-Layer Chromatography (HPTLC), Proton-Nuclear Magnetic Resonance (H-NMR) and MALDI-TOF. The antimicrobial activity of purified bacteriocin was determined using an agar-well assay against pathogenic organisms—Streptococcus mutans, Escherichia coli and Candida auris. The isolated peptide is characterized as a Class II bacteriocin with amino acid no. 8 (partial N-terminal sequence) and size 0.86 kDa (in accordance with RiPP nomenclature of bacteriocin classification). Peptides displayed potential antimicrobial effects with zones of inhibition against Escherichia coli at 0.2 cm, Streptococcus mutans at 0.3 cm and Candida albicans at 0.12 cm; p value = 0.00004 (p≤0.05). This is statistically significant. There is a current need to explore other such peptides and further apply them in pharmacological, food and biomedical sectors to strengthen their effectiveness against antimicrobial resistance. The current research data adds to the need to search for such alternate molecules.

Biofilm-driven infections represent a critical barrier to effective antimicrobial therapy, contributing significantly to the global burden of antibiotic resistance. Overcoming the limited biofilm penetration, rapid drug clearance, and suboptimal local antibiotic concentrations associated with conventional formulations remains a key therapeutic challenge. In this context, cefuroxime axetil-loaded polymeric nanoparticles based on Eudragit RS100 and RL100 were designed to enhance antibacterial and antibiofilm activity. Polymeric nanoparticle systems offer several advantages, including improved drug stability, enhanced penetration into the biofilm matrix, controlled and sustained drug release, and increased local drug concentration at the site of infection. Antibacterial activity was evaluated against clinically relevant biofilm-forming pathogens (Staphylococcus aureus ATCC 29213 and Escherichia coli ATCC 35218), revealing minimum inhibitory concentration (MIC) values ranging from 500 to 15.62 μg/mL. Importantly, the nanoparticulate system demonstrated pronounced antibiofilm efficacy across all tested concentrations. At 500 μg/mL, biofilm inhibition exceeded ≥94±1.48%, indicating near-complete suppression of biofilm formation. Remarkably, substantial inhibition ≥75±1.00% was retained even at the lowest concentration tested (15.62 μg/mL), underscoring the robustness of the formulation. The enhanced antibiofilm activity is likely driven by improved antibiotic diffusion into the biofilm matrix combined with sustained drug release characteristics of the Eudragit polymers, enabling prolonged antimicrobial exposure. Collectively, these findings position Eudragit-based cefuroxime axetil nanoparticles as a robust and promising platform to combat biofilm-associated infections and antimicrobial resistance. This approach highlights the potential of nanoparticle-mediated antibiotic delivery systems for next-generation therapeutic strategies targeting persistent infections.

This study was supported by Anadolu University Scientific Research Projects Commission under the grant no: YTT-2025-3144.

ONEPERÚ es una plataforma bioinformática desarrollada para la vigilancia genómica de bacterias resistentes a los antimicrobianos (RAM), enfocada en Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa y Acinetobacter baumannii. Integra datos fenotípicos, genómicos, epidemiológicos y clínicos provenientes de muestras humanas, ambientales y de animales de compañía, permitiendo analizar patrones de resistencia y apoyar decisiones en salud pública bajo el enfoque UNASALUD.

Durante el primer año (junio 2024 – junio 2025) se construyó la plataforma en su totalidad: arquitectura, diseño, mapas, tablas y módulo detallado de muestras. Asimismo, se recopilaron los requerimientos técnicos y funcionales, se amplió la capacidad para incluir cuatro microorganismos y se implementaron y validaron los procedimientos de carga y verificación de datos.

En el segundo año (julio 2025 – presente) se descargaron 115 genomas completos en formato FASTA desde el repositorio NCBI/GenBank, correspondientes a E. coli (30), K. pneumoniae (30), P. aeruginosa (30) y A. baumannii (25). Se desarrolló un script para automatizar la descarga y análisis genómico, registrándose variables como procedencia, accesión y características del genoma para determinar resistoma, genes de bombas de eflujo, viruloma, serotipos, filogrupos, secuenciotipos, plásmidos, fimbrias y k-locus.

Paralelamente, se secuenciaron 83 muestras mediante tecnología MinION (Oxford Nanopore), obtenidas de 75 aislados clínicos MDR, 4 de animales domésticos y 4 ambientales. A partir de archivos FASTQ, se ensamblaron genomas completos con alta calidad, lo que permitió caracterizar en profundidad el resistoma y viruloma, e identificar clones de alto riesgo como ST-486 de P. aeruginosa y ST11–KL111 de K. pneumoniae, reportados por primera vez en el país.

Finalmente, se proyecta continuar con el procesamiento genómico de 62 nuevos aislados ambientales y de animales de compañía mediante MinION, empleando scripts desarrollados para su análisis.