Introduction: Inappropriate use of antibiotics is one of the main drivers of antimicrobial resistance (AMR). The „Competency Framework for Education and Training on AMR for Health Workers” (WHO, 2018) defines the knowledge, attitudes, and practices required throughout the various domains of healthcare—including direct patient care, pharmaceutical services, laboratory diagnostics, and health management—to effectively reduce AMR-related burden.

Methods: A quantitative, single-center cross-sectional study was performed with purposive sampling among undergraduate biology and pharmacy students between 01/03/2021 and 01/02/2022. Data collection on AMR-related knowledge and practices was carried out using a 70-item, self-administered, online questionnaire based on the methodology of the Special Eurobarometer (EBM) 478 survey (European Commission, 2018). In addition, participants’ ability to recognize AMR-related terms and to identify pharmaceuticals (as antibiotic vs. non-antibiotic) was also assessed. Statistical analyses (descriptive statistics, Welch’s t-tests, and Pearson correlation) were performed using IBM SPSS 22.0. The study followed the Checklist for Reporting Results of Internet E-Surveys (CHERRIES) guidelines.

Results: Among participants (N=388), 20.4% reported antibiotic use in the past 12 months; 90.7% obtained antibiotics via medical prescription, of which only 18.8% were preceded by sampling or microbiological tests. Frequently reported indications for antibiotic use were sore throat (37.9%), cough (21.9%), and fever (21.6%). No relevant associations were identified between AMR-related knowledge and gender, academic performance, or type of study (p > 0.05). Strong, positive correlations were shown among AMR-related knowledge scores, the number of correctly recognized AMR-related terms, and the number of correctly identified pharmaceutical agents (r=0.644 and r=0.518, respectively; p<0.001).

Conclusions: Graduate biologists and pharmacists possess specialized competencies that are essential for mitigating AMR. Similarly to recent, population-based EBM surveys, our sample frequently reported antibiotic use for indications suggestive of antibiotic consumption in inappropriate indications. Our findings highlight the need for continuous evaluation and targeted strengthening of AMR-related content within undergraduate curricula.

Introduction: The consequences of antimicrobial resistance (AMR) are observed across different socio-economic strata and geographical regions; however, its key drivers and consequences are exacerbated by persisting economic and social inequalities. The AMR Quadripartite and the United Nations (UN) High-Level Political Forum have both empasized the importance of intersectoral actions addressing structual inequalities, as well as the need for sustained political commitment and policy predictability.

Methods: An ecological study was carried out based on secondary data collection, where the association between systemic (including community and hospital-based) antibiotic use (SABU) and thirty-three (N=33) indicators—corresponding to economic, developmental, political conditions, health systems status and inequality—for Hungary were assessed, over the period of 2010–2022. Data on SABU, expressed as defined daily doses per 1,000 inhabitants per day (DDD/1,000 inhabitants/day), were obtained from the European Centre for Disease Prevention and Control (ECDC) ESAC-Net database, while other indicators were sourced from the Hungarian Central Statistical Office (KSH), WHO Health for All, UN Human Development Reports, Our World in Data, and the World Bank DataBank databases, respectively. Statistical analyses (descriptive statistics, Spearman’s rho [ρ]) were conducted using jamovi version 2.4.5. The study followed the STrengthening the Reporting of OBservational studies in Epidemiology (STROBE) guidelines.

Results: SABU showed negative correlations with the mean number of years in education (rₛ=−0.688; p=0.009), number of nurses and midwives/10,000 inhabitants (rₛ=−0.564; p=0.044), gross domestic product (GDP) per capita (rₛ=−0.523; p=0.067), political stability index (rₛ=−0.529; p=0.062), gross national product (GNP) (rₛ=−0.502; p=0.081), and the Gini coefficient (rₛ=−0.484; p=0.094). Conversely, positive correlations were observed between SABU and the human rights index (rₛ=0.565; p=0.044) and with the proportion of the population living in poverty (rₛ=0.517; p=0.070).

Conclusions: Incorporating social stratifiers and the perspectives of intersectional is warranted in AMR-related data collection (e.g., surveillance), policy design and mitigation of AMR-related inequalities.

Introduction

Antibiotic resistance is one of the most pressing global health threats, diminishing the effectiveness of standard antimicrobial therapies and contributing to increased morbidity, mortality, and healthcare costs. In response to this challenge, ozone therapy has emerged as a potential adjunctive treatment against multidrug-resistant (MDR) bacterial infections. Notably, protocols endorsed by the Italian Scientific Society of Oxygen-Ozone Therapy (SIOOT) and clinical insights from Prof. Marianno Franzini, have advanced understanding of ozone dual mechanisms of action in infection control.

Methods

This review synthesizes data from experimental studies, modelling analyses, and clinical reports on the use of medical ozone in MDR infections. Mechanistic insights focus on two principal modes: (1) direct bactericidal activity of ozone and its reactive species through oxidative damage to microbial membranes and thiol groups, and (2) systemic immunomodulation via low-dose ozone exposure. The latter is mediated by secondary messengers such as 4-hydroxynonenal, which activate Nrf2 signaling, regulate inflammasomes, promote macrophage activity, and enhance mitochondrial function.

Results

Evidence indicates that ozone exerts rapid, broad-spectrum bactericidal effects largely independent of classical resistance mechanisms. When administered systemically within a hormetic dose range, ozone enhances host innate immunity through redox-sensitive signaling pathways and promotes tissue-level resilience. Modelling studies suggest synergistic effects when ozone is combined with antibiotics, accelerating pathogen clearance compared to antibiotic monotherapy. Clinically, ozone has demonstrated efficacy as an adjunct in treating chronic wounds and necrotizing infections caused by MDR pathogens.

Conclusions

Ozone therapy, when precisely dosed and integrated into standardized medical protocols, offers a compelling adjunctive approach in the management of antibiotic-resistant infections. Rather than replacing antibiotics, ozone augments their effectiveness and may reduce the selective pressure driving resistance. Future integration of ozone into antimicrobial stewardship programs could support more sustainable, system-based infection control strategies aligned with global health imperatives.

Bacterial biofilms represent a major challenge in the treatment of chronic infections due to their high tolerance to conventional antibiotics, notably seen in bone and joint infections. Bacteriophage therapy has re-emerged as a promising alternative or complementary strategy to address these limitations. In our lab, we investigate the use of bacteriophages encapsulated within a calcium phosphate-based delivery system known as biomimetic apatite powder, designed to mimic the mineral phase of bone tissue and ensure localized antimicrobial activity.

Phages specifically targeting Staphylococcus aureus and Escherichia coli were incorporated into this biomimetic apatite material using a precipitation process that preserves phage viability. The antibacterial efficacy of this system was evaluated against both planctonic infection and mature biofilms formed by abovementioned bacteriae. Our results demonstrate that phage-loaded biomimetic apatite significantly reduces viable bacterial counts and biofilm activity, highlighting the capacity of this encapsulation system to deliver active phages in a sustained and localized manner. Planctonic culture of E. coli and S. aureus were completely eradicated (0 CFU, 7 log reduction) after 2 hours of incubation with the phage loaded material. Biofilm culture of S.aureus were almost completely eradicated ( ∼6,4 CFU, 5 log reduction) after 5 days of incubation with the lyophiliosed phage-loaded material but not with the lyophilised version (∼2,8.10³ CFU, 3 log reduction).

Furthermore, we explored the synergistic effects between phage therapy and conventional antibiotics. Combined treatments resulted in enhanced biofilm disruption and bacterial eradication compared to phages or antibiotics alone, suggesting a strong synergistic interaction. This combined approach may help overcome antibiotic tolerance associated with biofilms while reducing the required antibiotic doses as shown in other studies.

Overall, our findings support the potential of biomimetic apatite powders as an effective delivery platform for bacteriophages and as a promising strategy for the treatment of biofilm-associated infections, particularly in the context of bone and joint diseases.

The Global Antibiotic Research & Development Partnership (GARDP) accelerates the development and access of treatments for drug-resistant infections. Together with public, private, and non-profit partners, GARDP works in partnership to develop new antibiotics and expand access to them. Through our Discovery & Exploratory Research (DER) programme, GARDP focuses on the search for novel molecules and new bacterial targets against WHO-critical Gram-negative bacterial pathogens. This work contributes to the global antibiotic R&D pipeline, aiming to create a steady stream of promising new candidate drugs. One research area of the DER programme focuses on the discovery of potentiating compounds that can restore the activity of clinical antibiotics lost to resistance, such as efflux inhibitors.

Many antibiotics are substrates of bacterial efflux pumps, and modifications to the structure or overexpression of efflux pumps are an important resistance mechanism used by many multidrug-resistant bacteria. Therefore, chemical inhibition of bacterial efflux to revitalize existing antibiotics is considered a promising approach for antimicrobial chemotherapy. We recently provided an overview of clinically relevant multidrug resistance efflux pumps in Gram-negative bacteria and further described over 50 efflux inhibitors that target such systems. Following our medicinal chemistry and microbiology review, we selected promising efflux inhibitors for benchmarking, using a K. pneumoniae ∆ramR mutant with decreased susceptibility to antibiotics effluxed by the AcrAB-TolC system. The results will determine if these compounds are viable starting points for hit-to-lead projects focused on efflux inhibition in Gram-negative bacteria.

Abstract

Antimicrobial resistance (AMR) poses a significant global health threat, ranking among the top ten threats to humanity according to the World Health Organization [1]. The rise of multidrug-resistant (MDR), extensively drug-resistant (XDR), and pan-drug-resistant (PDR) pathogens is linked to their increasing insensitivity to traditional antibiotics [2]. Antimicrobial peptides (AMPs) offer a promising solution to combat drug-resistant infections [3]. These peptides, which possess cationic and hydrophobic regions, can disrupt bacterial cell membranes, leading to bacterial death [4]. However, challenges such as instability, high costs, and toxicity to normal cells must be addressed. Metallacarboranes are stable compounds consisting of one or more boron hydride clusters coordinating a metal cation. They possess unique structures and properties that make them a promising platform for the development of new antimicrobial agents [5].

In our studies, we synthesized a series of metallacarborane–ultrashort cationic peptide conjugates and comprehensively characterized their physicochemical properties. Their antimicrobial activity was evaluated by determination of minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) against representative Gram‑positive and Gram‑negative bacteria, demonstrating broad‑spectrum and predominantly bactericidal effects at low micromolar levels. In parallel, cytotoxicity and hemolysis assays were performed, enabling calculation of selectivity indices that relate MIC/MBC values to toxicity thresholds and thus substantiate the favorable therapeutic window of these conjugates.

ACKNOWLEDGMENTS

The study was supported by a grant from the National Science Centre, Poland (grant number 2023/51/D/NZ7/02609).

REFERENCES

1] Kumar, S. EClinicalMedicine 2021, 41, 101221.

[2] Pulingam, T.; Parumasivam, T.; Gazzali, A. M.; Sulaiman, A. M.; Chee, J. Y.; Lakshmanan, M.; et al. Eur. J. Pharm. Sci. 2022, 170, 106103.

[3] Bucataru, C.; Ciobanasu, C. Microbiol. Res. 2024, 127822.

[4] Matthyssen, T.; Li, W.; Holden, J. A.; Lenzo, J. C.; Hadjigol, S.; O’Brien-Simpson, N. M. Front. Chem. 2022, 9, 795433.

[5] Fink, K.; Uchman, M. Coord. Chem. Rev. 2021, 477, 214940

Human adenovirus and norovirus are environmentally stable, non-enveloped viruses that remain major causes of infections worldwide. Despite their clinical significance, effective antiviral therapeutics targeting these pathogens remain limited. Previous in vitro studies demonstrated that the antimicrobial peptides Mel4 and lactoferricin (LFc) possess antiviral activity against adenovirus type 5 (HAdV-5) and murine norovirus type 1 (MNV-1) and do not cause toxicity at their antiviral concentrations to the host mammalian cells. The ID₅₀ of Mel4 was lower than that of LFc. However, although some data indicated that the mode of action was directed at the capsids of these viruses, the molecular basis of the antiviral activity was not fully elucidated. In this study, in silico approaches were employed, such as HDOCK docking and molecular dynamics (MD) simulations, to understand how Mel4 and LFc interact with key capsid proteins. Structural models of Mel4 and LFc were generated using AlphaFold3, and their physicochemical properties were characterised. Docking analyses against the HAdV-5 hexon, penton base, and fibre knob proteins, as well as the MNV-1 VP1 capsid protein, revealed consistently stronger binding affinities for Mel4 compared with LFc. MD simulations (100 ns) further demonstrated that Mel4–capsid complexes exhibited lower root mean square deviation values, reduced interfacial flexibility, and greater structural compactness relative to LFc complexes, indicating stronger and more stable binding over time. Increased solvent accessibility of Mel4 also suggested its greater interaction potential with viral surfaces. Together, these findings provide a detailed mechanistic framework supporting the superior antiviral activity of Mel4 and highlight specific capsid regions that might serve as targets for peptide-based antiviral development. This study has enhanced the understanding of peptide–virus interactions at the molecular level and offers valuable insights for the rational design of next-generation antivirals against environmentally resilient, non-enveloped viruses such as adenovirus and norovirus.

Antibiotic resistance (AMR) is a critical issue within the "One Health" approach, which encompasses human and animal health, agriculture, and environmental waste management. Fresh vegetables, which are frequently consumed raw, may contribute to the spread and transmission of AMR. Therefore, the aim of this research was to study the microbiological quality and prevalence of ARB and ARG in fresh vegetables.

Twenty-one samples were purchased directly from organic farms in Valencia (Spain), and two soil samples were taken from the cultivation sites. Samples were examined for total viable bacteria, coliforms, Escherichia coli, Listeria monocytogenes and Salmonella spp. Isolation was performed onto culture media supplemented with cefotaxime and meropenem.

A total of 122 strains were isolated. A total of 24 Gram-negative isolates were selected and identified by biochemical tests and sequencing of 16S rRNA. Their sensitivity to 15 antibiotics was determined, and PCR assays were performed for main expanded spectrum beta-lactamase (ESBL), carbapenemase- and plasmid-mediated quinolone resistance genes, both for the samples and for the isolates.

Overall, 96% of the samples showed total viable bacteria levels over the accepted 10(5) cfu/g standard limit, and 87% showed coliforms levels over the accepted 10(4) cfu/g standard limit. In two samples (9%), E. coli levels were not acceptable. Salmonella and L. monocytogenes were not detected.

Seventy-nine percent of the isolates were resistant to at least one antibiotic, and 50% were ESBL productors. PCR showed the presence of bla_CMY-2 (13%), bla_TEM (8%), bla_SHV (4%), bla_OXA-48 (17%), bla_KPC (8%), bla_IMP (4%), qnrA (17%), qnrB (21%) and qnrS (8%) genes.

The detection of carbapenems resistance genes is particularly concerning because it poses a serious risk for immunocompromised patients. Thus, fresh organic vegetables harboring ARB and ARG constitute a potential risk to consumers. Further studies must be done to detect ARG and how they propagate in non-medical environments.

Staphylococcus aureus causes difficult-to-treat infections with current antibiotics, often linked to resistant biofilms. Honey has the potential to act as an antibiotic due to its acidity and high sugar content. Mechanisms by which honey acts as an antibiotic are unclear. We hypothesize that exposing S. aureus cells to Manuka honey induces physiochemical changes to cells in a concentration-dependent manner. To test our hypotheses, Atomic Force Microscopy (AFM) was used to image cells and quantify their adhesion to a model surface. The Hertz model of contact mechanics was used to estimate the Young’s moduli of the cells. S. aureus was exposed to 10%, 20%, 30% (MIC), and 40% honey for 2 hours. Untreated cells were the negative control. AFM was performed at room temperature on 3 cells per slide, 3 slides per culture, and 3 cultures per condition. Exposure of cells to 10%, 20% and 30% honey for 5 hours resulted in 0%, 45%, and 100% killing of cells, respectively. When cells were untreated, adhesion forces, energies, and elasticities were 17.79±39.72 pN, 4.87±5.06 aJ, and 221.19±100.22 kPa, respectively. Exposure to 10% honey enhanced adhesion forces and energies to 51.41±38.91 pN and 11.86±9.88 aJ, respectively, and did not affect elasticity. Exposure to 20% honey increased adhesion forces and energies further to 102.09±101.04 pN and 14.00±12.91 aJ, respectively, and did not affect elasticity. At 40% honey (above MIC), the adhesion forces and energies reached their maximum values at 356.22±276.98 pN and 81.07±66.37 aJ, respectively, while the elasticity dropped by 70% to 64.32±42.91 kPa. Cells were intact under all conditions investigated. The drop of elasticity could be the result of detachment of surface molecules, while increased adhesion may be associated with increased flexibility of surface molecules. Our results suggest the excellent potential for honey to be used as an antibacterial agent.

The widespread use of antibiotics as growth promoters in poultry production has contributed to the emergence of antimicrobial resistance and the dissemination of zoonotic pathogens, underscoring the urgent need for effective non-antibiotic alternatives. This study aimed to evaluate the in vitro antibacterial activity of oregano (Origanum vulgare) essential oil (OEO), rosemary (Salvia Rosmarinus) essential oil (REO), and their combination (OR) against Escherichia coli and Salmonella typhimurium strains of poultry origin.

Minimum inhibitory and bactericidal concentrations were determined, and the optimal OR combination was selected using checkerboard assays. Time–kill curves were performed using ATCC reference strains (E. coli ATCC 25922 and S. Typhimurium ATCC 14028), as well as poultry field isolates (n = 2 per species), exposed to individual essential oils and their combination at different multiples of the MIC, under pH conditions simulating the avian gastrointestinal tract (pH 5.0 and 7.4). Bacterial viability was monitored over time based on viable cell counts (CFU/mL), and antibacterial activity was quantified using the antibacterial effect index (E). Results were interpreted according to established cut-off values for bacteriostatic activity (E = 0), bactericidal activity (E ≤ −3), and virtual eradication (E ≤ −4).

Both essential oils exhibited rapid and concentration-dependent antibacterial activity. At 2× and 4× MIC, OEO and REO induced immediate bacterial killing, achieving virtual eradication (E ≤ −4) within 10–20 minutes of exposure. The OR combination consistently enhanced antibacterial activity, reaching bactericidal and eradication thresholds faster and at lower concentrations compared to individual oils. Comparable killing dynamics were observed across pH conditions and between reference strains and poultry field isolates for both E. coli and S. typhimurium.

These findings demonstrate the potent and rapid antimicrobial action of OEO and REO, particularly when used in combination, supporting their potential application as non-antibiotic growth-promoting alternatives in poultry production within a One Health framework.