Background/Objectives: Non-HACEK Gram-Negative Infective Endocarditis (NHGNIE) is a rare but increasingly recognized condition associated with high morbidity and mortality. Its incidence is rising among people who inject drugs (PWID), patients with prosthetic valves or cardiac devices, and those with significant healthcare exposure. We aimed to provide a comprehensive review of the epidemiology, pathogenesis, diagnosis, clinical features, and management of NHGNIE. Methods: We conducted a narrative synthesis of published cohort studies, case series, guideline documents, and recent registry data addressing NHGNIE. Evidence was extracted and critically appraised with emphasis on epidemiological patterns, microbial etiology, diagnostic frameworks, therapeutic strategies, and outcomes. Special focus was given to pathogen-specific differences and the impact of antimicrobial resistance. Results: NHGNIE accounts for approximately 1.5–10.7% of IE cases worldwide, with marked geographical variability. Pseudomonas aeruginosa, Serratia marcescens, Klebsiella pneumoniae, and Escherichia coli are the predominant pathogens, with clinical profiles differing between younger, PWID-based populations and older, comorbidity-affected cohorts. Advances in molecular diagnostics and imaging have improved case identification, though pathogen-specific diagnostic performance remains limited. Outcomes are poor, with in-hospital mortality up to 41%. Antimicrobial therapy is complicated by biofilm formation and potential for multidrug resistance; evidence for combination therapy versus monotherapy is conflicting. Surgical intervention appears to improve outcomes when performed according to guideline-based indications, but results are heterogeneous across studies. Conclusions: NHGNIE is a clinically significant form of IE with complex epidemiology, diagnostic challenges, and limited evidence to guide treatment. Effective management requires individualized care coordinated within a multidisciplinary “endocarditis team”.

Background/Objectives: This study aimed to analyze the consumption of systemic antibacterials (WHO ATC J01) in primary care in Kazakhstan from 2017 to 2024 and to estimate the potential contribution of different primary care providers to the use of “Watch” and “Reserve” antibiotics. Methods: The Global Antimicrobial Resistance and Use Surveillance System methodology was applied to calculate defined daily doses per 1000 inhabitants per day (DID) for ATC J01 overall, as well as at ATC levels 3 and 5. Forecast modeling was performed using ARIMA (0,1,0), ARIMA (0,0,0), and Simple models to project trends through 2030. Pearson’s correlation was computed for each category of primary healthcare provider and each chemical agent belonging to the “Watch” and “Reserve” groups. Results: Antibiotic consumption increased by approximately 1.5 times in 2024 after remaining relatively stable from 2017 to 2023. The forecasts suggest that the share of “Access” group antibiotics will continue to decline, reaching 33.71% by 2030, whereas the share of “Watch” group antibiotics is expected to increase proportionally, reaching 65.73% by 2030. The per capita densities of primary care providers correlate with the consumption of certain “Watch” and “Reserve” group antibiotics. Conclusions: This study demonstrates that the primary care sector in Kazakhstan is characterized by a disproportionately high and increasing use of “Watch” antibiotics, a rising trend in “Reserve” antibiotic consumption, and a declining share of “Access” antibiotics. These findings highlight a need to prioritize stewardship interventions that target the reduction in “Watch” and “Reserve” antibiotics, while promoting the use of “Access” antibiotics.

Colistin resistance, driven by chromosomal mutations and the spread of plasmid-mediated MCR genes, has emerged as a critical challenge in combating multidrug-resistant Gram-negative bacteria. This resistance compromises the efficacy of colistin, leading to higher treatment failure rates, prolonged hospitalizations, and increased mortality. Recent studies have highlighted key mechanisms, including lipid A modifications, that enable bacteria to evade colistin’s effects. The global spread of MCR genes exacerbates the issue, underlining the need for improved diagnostics and rapid detection of resistant strains to prevent adverse patient outcomes. To combat this growing threat, a multifaceted approach is essential, involving enhanced antimicrobial stewardship, stricter infection control measures, and continued research into alternative therapies and diagnostic methods. Collaborative efforts from researchers, healthcare providers, policymakers, and the pharmaceutical industry are crucial to preserving colistin’s effectiveness and mitigating the broader impact on public health.

Background: As of 2022, 80% of all documented microbial infections are biofilm-associated: communities of microorganisms adhered to a surface and enclosed in a complex extracellular polymeric substance (EPS). The EPS acts as a physical barrier protecting the bacteria from antimicrobial agents and host immune responses. To combat this hurdle, the application of glycoside hydrolases (GH) has been investigated due to their ability to cleave particular structural polysaccharides within the EPS, thus breaking down the protective barrier and improving antibiotic clearance. While various studies demonstrate the capacity of GHs to improve antibiotic efficacy against biofilms in combination, there is clear differential success between these treatments depending on the GH and antibiotic chosen. Due to the overlap of GH targets and antibiotic structures, it is imperative to ensure that the antibiotics in combinatorial treatments are not degraded by the GH. Methods: This study aimed to screen the GH α-amylase produced from Aspergillus oryzae (AO) and Bacillus subtilis (BS), combined with various antibiotics from different classes, charges, and mode of actions by determining MICs. against the bacterium Pseudomonas aeruginosa (PA) of 6 antibiotics with or without α-amylase and treat 2-day PA biofilms with antibiotics with or without GHs. Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) stability assays and Differential Scanning Fluorimetry (DSF) were conducted to determine antibiotic and GH degradation as well as antibiotic sequestration. Results: Increased MICs in the presence of GHs as well as decreased antibiotic clearance against 2-day biofilms were suggestive of antibiotic degradation. LC-MS/MS stability assays of tetracycline and ciprofloxacin in the presence and absence of α-amylase further demonstrated the α-amylase-mediated antibiotic sequestration. Differential scanning fluorimetry (DSF) assays confirmed α-amylase-antibiotic interactions. Conclusions: This study suggests that α-amylase is capable of degrading and sequestering a variety of antibiotics, and the degree to which these phenomena occur varies depending upon the source of the GH. As a potential treatment for biofilm-associated infections, it is imperative that the GH + antibiotic combinations are determined compatible prior to clinical use.

Introduction: The rise in hospital-acquired pneumonia (HAP) due to antibiotic-resistant bacteria is increasing morbidity, mortality, and inappropriate empirical antibiotic use. This prospective research aimed to evaluate the performance of a real-time polymerase chain reaction (PCR) assay for detecting causative microorganisms and antibiotic-resistance genes from respiratory specimens compared to traditional methods. Additionally, we aimed to determine the molecular epidemiology of antibiotic resistance genes among HAP patients at The University of Jordan hospital. Methods: Lower respiratory tract samples were collected from HAP patients, including those with ventilator-associated pneumonia (VAP), between May 2024 and October 2024. Clinical data from the medical files were used to collect and analyze demographic and clinical information, including clinical outcomes. Real-time PCR was run to detect causative microbes and antibiotic resistance genes. Results: Among 83 HAP patients (median age 63, 61.45% male), 48.15% died. Culture identified Klebsiella (25.53%), Acinetobacter (22.34%), and Candida (24.47%) as the most common pathogens, while qPCR showed higher detection rates, including for A. baumannii (62.20%, p = 0.02) and K. pneumoniae (45.12%, p < 0.001). Carbapenem resistance was high; A. baumannii showed 100% resistance to most antibiotics except colistin (92.31%). The resistance genes ndm (60%) and oxa-48 (58.46%) were frequently detected and significantly associated with phenotypic resistance (p < 0.001). The qPCR identified resistance genes in all carbapenem-resistant cases. No gene significantly predicted mortality. Conclusions: Real-time PCR diagnostic technique combined with epidemiology of antibiotic resistance genes data may be a rapid and effective tool to improve HAP management. Large, multicenter studies are needed in the future to validate the performance of real-time PCR in HAP diagnosis, and appropriate management is also required.

Antimicrobial resistance (AMR) is a growing threat to global health, accelerated by the widespread inappropriate use of antibiotics. Although educational initiatives have been launched worldwide, there is little evidence on how younger generations in high-income countries (HICs) understand and address AMR. Addressing the AMR crisis requires proactive education of younger generations, including children, adolescents, and young adults, who will shape future healthcare practices. This review analyzes existing research on AMR literacy among these age groups in HICs, as knowledge gaps and risky behaviors persist even in HICs, despite their strong education and health infrastructures. The purpose of this review is to examine the knowledge, attitudes, and behaviors related to antibiotic use and antibiotic resistance in younger generations while identifying effective educational interventions. Methods: We performed a comprehensive literature search in PubMed until June 2025, followed by AI-assisted screening (Claude 4.0 Sonnet) and a manual review. The search strategy combined terms from the areas of health literacy, antibiotics, antibiotic resistance/AMR, and young populations. Studies in HICs that examined the younger generation’s knowledge about antibiotics and AMR, analyzed their attitudes or behavior toward them, or evaluated relevant educational interventions were included. Data were synthesized thematically across all included studies. Results: Nineteen studies from 11 HICs were included, including thirteen cross-sectional surveys and six educational intervention studies. The results showed that misconceptions about how antibiotics work are still very common. Several of those asked (22–80%) incorrectly stated that resistance develops in the human body and not in bacteria. Many (26–77%) mistakenly agreed with the statement that antibiotics treat viral infections. Concerning behaviors included high rates of self-medication, non-adherence to treatment, and unsafe storage practices. Several authors propose an amendment of curricula. Educational interventions, particularly gamification and peer education approaches, showed improvements in knowledge and sustained learning outcomes. Conclusions: Knowledge of AMR among young people in HICs is still inadequate, despite educational advantages. Most existing studies focus on college students, while children and adolescents, crucial groups for early prevention, are underrepresented. Targeted, age-appropriate education employing interactive methods represents an evidence-based strategy to improve antibiotic use behavior and support global AMR control efforts.

This study investigated the molecular and phenotypic characteristics of antimicrobial resistance in Escherichia coli isolates recovered from the ceca of healthy broilers in Tunisia. A total of 111 E. coli isolates were obtained from chicken samples collected at slaughterhouses and cultured on cefotaxime-supplemented MacConkey agar. All isolates exhibited a multidrug-resistant (MDR) phenotype, and 72.1% were confirmed as extended-spectrum β-lactamase (ESBL) producers. The most frequent β-lactamase gene was blaCTX-M-G1, followed by blaTEM and blaSHV. Carbapenem resistance genes (blaOXA-48 and blaIMP) were detected in 12.6% and 6.3% of isolates, respectively, while six isolates harbored the colistin resistance gene mcr-1. Among the tested virulence genes, fimH, traT, and iutA were the most prevalent, detected in over 70% of isolates. Class 1 integrons were present in 83% of isolates, and class 2 integrons in 39.6%, with gene cassettes encoding resistance to trimethoprim (dfrA) and streptomycin (aadA). These findings highlight the widespread presence of MDR and ESBL-producing E. coli strains with virulence traits and integrons in poultry, underscoring the risk of transmission to humans. This study provides essential data supporting the implementation of integrated surveillance strategies in line with the One Health approach.

Background/Objectives: This report documents the first confirmed case in Romania of fatal pneumonia and septicemia in a domestic rabbit caused by multidrug-resistant Escherichia coli, highlighting both its pathogenic potential and One Health implications. Case Study: An 8-month-old male German Giant Spotted rabbit raised on a rural farm under poor husbandry conditions developed acute respiratory distress and died within 48 h. Post-mortem examination revealed severe pulmonary congestion, tracheal inflammation, serofibrinous pericarditis, and systemic vascular lesions. Bacteriological analysis confirmed E. coli from lung, trachea, and bone marrow samples. The isolate demonstrated strong Congo red binding, was confirmed by MALDI-TOF mass spectrometry, and showed resistance to beta-lactams, fluoroquinolones, tetracyclines, sulfonamides, macrolides, and phenicols, remaining susceptible only to aminoglycosides. PCR screening identified virulence genes (fimH, papC, iutA, ompA) linked to adhesion, immune evasion, and iron acquisition, with potential for horizontal gene transfer. Conclusions: This first documented case in Romania emphasizes the clinical threat posed by multidrug-resistant E. coli in rabbits and the importance of early diagnosis, improved biosecurity, and responsible antimicrobial use. The zoonotic and environmental risks in backyard farming underscore the urgent need for integrated surveillance. Alternative control strategies, including phage therapy and probiotics, should be explored to reduce reliance on conventional antibiotics.

Background/Objectives: Antimicrobial stewardship (AMS) interventions are critical to reducing inappropriate antibiotic prescribing for acute respiratory infections (ARIs) in primary care and mitigating antimicrobial resistance (AMR). While interventions are routinely employed in hospitals, implementation in general practice is nascent. This qualitative study, part of the OPTIMAS-GP project, explored determinants influencing the implementation of evidence-based AMS strategies in Australian general practice. Methods: Using Experience-Based Co-Design, three rounds of online focus groups were conducted with ten healthcare professionals (GPs, pharmacists, microbiologist, practice staff) and ten adult patients who had experienced ARI management in primary care. Participants discussed the feasibility and acceptability of AMS interventions: shared decision-making (SDM) tools, delayed prescribing (DP) and point-of-care testing (PoCT) for C-reactive protein (CRP). Results: Thematic analysis of focus group transcriptions identified four interrelated themes: ‘Patient acceptance and engagement’, ‘Practising within a system’, ‘Prescribing stewardship’, and ‘Diagnostic stewardship’. Patient engagement was dependent upon expectations, trust, and personalised care, while systemic factors such as continuity of care, practice culture, and resource availability influenced implementation. DP was viewed as a pragmatic but potentially confusing strategy, requiring clear patient guidance and interprofessional collaboration. SDM tools were conceptually supported but challenged by time constraints and poor health literacy. PoCT-CRP was cautiously welcomed for selective use, with concerns expressed about workflow integration and overreliance on testing. Findings were mapped to the Capability, Opportunity, Motivation-Behaviour (COM-B) and Theoretical Domains Framework (TDF) to identify behavioural determinants and inform future implementation strategies. Recommendations include co-designing patient-centred AMS tools with clear instructions and red flags, enhancing GP-pharmacist collaboration, and addressing barriers to PoCT integration. Conclusions: These insights highlight the complexity of implementing AMS interventions in general practice and underscore the need for tailored, system-supported approaches to optimise antibiotic use and reduce AMR.

Antimicrobial resistance (AMR) has emerged as one of the greatest public health challenges in the last decades [...].