Antibiotics

23 pages, 2061 KB

Open AccessReview

Artificial Intelligence and the Discovery of Antibiotics: Reinventing with Opportunities, Challenges, and Clinical Translation

by Bharat Kumar Reddy Sanapalli, Shrestha Palit, Ashwini Deshpande, Ramya Tokala, Dilep Kumar Sigalapalli and Vidyasrilekha Sanapalli

Antibiotics 2026, 15(2), 233; https://doi.org/10.3390/antibiotics15020233 - 23 Feb 2026

Cited by 9 | Viewed by 3739

Abstract

Background: The outbreak and spreading of antimicrobial resistance (AMR) in a very short time has made most of the old-fashioned antibiotics ineffective, and thus new therapeutic substances have to be developed. The traditional methods of antibiotics discovery are defined by long periods of [...] Read more.

Background: The outbreak and spreading of antimicrobial resistance (AMR) in a very short time has made most of the old-fashioned antibiotics ineffective, and thus new therapeutic substances have to be developed. The traditional methods of antibiotics discovery are defined by long periods of time, high levels of expenditure, and high rates of failure, which contributes to the necessity of new approaches. Artificial intelligence (AI) has become a disruptive technology that can be used to accelerate and optimize various steps of antibiotic discovery, such as target detection and virtual screening, new molecular design, and early-stage testing. Methods: This review provides an in-depth discussion of the role of AI methodologies in the form of machine learning, deep learning, natural language processing, and generative models in the discovery of small-molecule antibiotics and antimicrobial peptides (AMPs). The major areas that are discussed include virtual screening, pharmacokinetics optimization, resistance mechanism prediction, and AMPs design, which is accompanied by relevant case studies, including the AI-based discovery of Abaucin. Results: The article highlights how AI can be used in a synergistic relationship with synthetic biology, nanotechnology, and multi-omics data as a core component in the next generation of antimicrobial approaches, such as personalized therapy and predictive stewardship. The existing issues, i.e., the lack of data, bias in algorithms, and the translational divide between research and clinical use, are addressed, as well as suggested measures of responsible, collaborative, and ethical AI use. Conclusions: The combination of computational innovation with experimentation validation, AI-driven antibiotic discovery paves the way for a potent and scalable approach in addressing the rising threat of AMR. Full article

(This article belongs to the Special Issue Evaluation of Emerging Antimicrobials, 2nd Edition)

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21 pages, 1779 KB

Open AccessArticle

Clinical and Microbiological Profile of Hospital-Acquired and Ventilator-Associated Pneumonia in Critically Ill Patients: A Retrospective Observational Study

by Mihnea Miron, Anca Irina Ristescu, Mihaela Blaj, Diana Gabriela Iosep, Alexandru-Florinel Oancea, Gabriel Iosep, Radu Crișan-Dabija, Daniela Diculencu, Costin Damian and Mihaela Cătălina Luca

Antibiotics 2026, 15(2), 232; https://doi.org/10.3390/antibiotics15020232 - 22 Feb 2026

Cited by 4 | Viewed by 2221

Abstract

Background/Objectives: Severe respiratory infections remain a major cause of morbidity and mortality in critically ill patients admitted to an intensive care unit (ICU), particularly in the context of increasing antimicrobial resistance (AMR). This study aimed to describe the clinical, microbiological and resistance [...] Read more.

Background/Objectives: Severe respiratory infections remain a major cause of morbidity and mortality in critically ill patients admitted to an intensive care unit (ICU), particularly in the context of increasing antimicrobial resistance (AMR). This study aimed to describe the clinical, microbiological and resistance profiles of ICU patients diagnosed with hospital-acquired or ventilator-associated pneumonia. Methods: We conducted a retrospective, single-center observational study including adult ICU patients admitted between January and December 2025, with clinically significant positive endotracheal aspirates. Clinical severity scores (APACHE II, SOFA, SOFA-2), inflammatory biomarkers (neutrophil-to-lymphocyte ratio—NLR, platelets-to-lymphocyte ratio—PLR, C-reactive protein—CRP), microbiological findings, antimicrobial resistance patterns and ICU-related outcomes were analyzed. Results: Out of the 606 endotracheal aspirates collected, 76 (12.5%) were culture-positive and 62 (10.2%) patients met the final inclusion criteria. Ventilator-associated pneumonia accounted for 90% of infections, 25 episodes (44.6%) being classified as early-onset and 31 cases (55.4%) as late-onset, without significant differences in bacterial distribution between the two subtypes. In total, 85.5% of infections were monomicrobial, with Gram-negative bacteria representing 76% of isolates. Acinetobacter baumannii and Pseudomonas aeruginosa were the most frequently isolated pathogens, with high resistance rates. Acute kidney injury occurred in 25.8% of patients and was associated with higher APACHE II, SOFA, and SOFA-2 scores. Conclusions: Severe respiratory infections in critically ill patients were predominantly caused by Gram-negative, frequently drug-resistant pathogens and were associated with high disease severity and poor outcomes. These findings provide insight into the local epidemiology and antimicrobial resistance patterns of severe respiratory infections in critically ill patients. Full article

(This article belongs to the Special Issue Epidemiology and Treatment of Microbial Infections in Respiratory Diseases)

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11 pages, 228 KB

Open AccessArticle

Use of RESERVE-Antibiotics in Newborns: Clinical Experience of Two NICUs in the Metropolitan Area of Palermo

by Veronica Notarbartolo, Deborah Bacile, Bintu Ayla Badiane, Agnese Lo Leggio, Vita Maria Angileri, Vincenzo Duca and Mario Giuffré

Antibiotics 2026, 15(2), 231; https://doi.org/10.3390/antibiotics15020231 - 21 Feb 2026

Cited by 1 | Viewed by 1124

Abstract

Background: The increasingly indiscriminate use of antibiotic therapy in the neonatal period has led to the emergence of multidrug-resistant organisms (MDROs), which are responsible for sepsis that is increasingly difficult to treat and associated with high morbidity and mortality. Increasingly frequently, in [...] Read more.

Background: The increasingly indiscriminate use of antibiotic therapy in the neonatal period has led to the emergence of multidrug-resistant organisms (MDROs), which are responsible for sepsis that is increasingly difficult to treat and associated with high morbidity and mortality. Increasingly frequently, in neonatal intensive care units (NICUs), it is necessary to use last-generation antibiotics belonging to the RESERVE group according to the current classification of the World Health Organization (WHO). Methods: Among these drugs, ceftazidime-avibactam, ceftolozane-tazobactam and meropenem-vaborbactam are increasingly used in infections caused by Enterobacterales (i.e., E. cloacae complex, Klebsiella spp.), which are often responsible for late-onset sepsis (LOS) in newborns, especially in preterms. Results: Here, we present the experience of four newborn patients in the city of Palermo, treated over a period of 3 years. Conclusions: The comparison between different diagnostic–therapeutic management approaches and a review of the most recent literature can contribute to identifying more standardized pharmacological schemes, especially in the neonatal period, where scientific evidence about this topic is still very limited. Full article

(This article belongs to the Special Issue Neonatal Infection: Antibiotics for Prevention and Treatment)

17 pages, 570 KB

Open AccessArticle

Substandard and Falsified Antibiotics Seized in Belgium: Quality Control Analysis Reveals High Prevalence of WHO Watch List Molecules and Bioavailability Non-Compliance

by Celine Vanhee, Cloë Degrève, Michael Canfyn, Niels Boschmans, Bram Jacobs, Koenraad Van Hoorde, Eric Deconinck, Marie Willocx, Hans Van der Meersch and Bart Ceyssens

Antibiotics 2026, 15(2), 230; https://doi.org/10.3390/antibiotics15020230 - 21 Feb 2026

Cited by 1 | Viewed by 1194

Abstract

Background: Antimicrobial resistance (AMR) poses a critical global public health challenge requiring comprehensive intervention strategies, including robust antibiotic stewardship programs. The European Commission’s 2017 One Health Action Plan against AMR established guidelines based on WHO’s AWaRe classification system, which categorizes antibiotics into Access, [...] Read more.

Background: Antimicrobial resistance (AMR) poses a critical global public health challenge requiring comprehensive intervention strategies, including robust antibiotic stewardship programs. The European Commission’s 2017 One Health Action Plan against AMR established guidelines based on WHO’s AWaRe classification system, which categorizes antibiotics into Access, Watch, and Reserve groups to promote prudent use. However, the proliferation of substandard and falsified (SF) medical products increasingly undermines these stewardship efforts, with European regulatory agencies, including Belgium’s Federal Agency for Medicines and Health Products (FAMHP), reporting rising seizures of SF antibiotics. Objective: To assess the pharmaceutical quality of confiscated antibiotic samples and evaluate their potential contribution to AMR development. Methods: We conducted comprehensive pharmaceutical quality control testing on 40 SF antibiotic samples seized by the FAMHP between early 2024 and late 2025. Results: The analysis revealed significant deficiencies: 35% of samples contained antibiotics listed on international watch lists, while nearly 43% failed quality control testing. Dissolution defects represented the predominant failure mode, accounting for 29% of all samples tested. These defects can severely compromise drug bioavailability, clinical efficiency, and expose bacterial populations to sub-lethal concentrations of active pharmaceutical ingredients—a recognized driver of resistance development. Conclusions: Many seized samples appeared to be unregistered/unlicensed medicines that, while prohibited in the EU, may circulate legally elsewhere. This transnational dimension highlights how substandard products threaten global AMR control initiatives beyond individual patient safety. These findings underscore the urgent need to raise more awareness within the EU and for enhanced pharmaceutical quality assurance systems and much more international regulatory cooperation, particularly in low- and middle-income countries where such products circulate more readily. Full article

(This article belongs to the Section Antibiotics Use and Antimicrobial Stewardship)

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17 pages, 694 KB

Open AccessArticle

Prospective Evaluation of ESBL Risk Factors and Appropriateness of Empirical Therapy in Hospitalized Patients with Community-Onset Pyelonephritis

by Gülşah Gelişigüzel, Şerife Altun Demircan, Murat Aysin, Esra Kaya Kılıç, Serap Yağcı, Sami Kınıklı and Rukiye Berkem

Antibiotics 2026, 15(2), 229; https://doi.org/10.3390/antibiotics15020229 - 20 Feb 2026

Cited by 1 | Viewed by 989

Abstract

Background/Objectives: The rising prevalence of extended-spectrum beta-lactamase (ESBL)-producing pathogens has emerged as a significant challenge in the treatment of pyelonephritis. This study aims to determine the frequency of ESBL-producing agents in hospitalized patients with pyelonephritis, identify associated risk factors, and assess the [...] Read more.

Background/Objectives: The rising prevalence of extended-spectrum beta-lactamase (ESBL)-producing pathogens has emerged as a significant challenge in the treatment of pyelonephritis. This study aims to determine the frequency of ESBL-producing agents in hospitalized patients with pyelonephritis, identify associated risk factors, and assess the appropriateness of empirical antimicrobial therapy. Methods: This prospective study included patients hospitalized with pyelonephritis in the Infectious Diseases Clinic of Ankara Training and Research Hospital between 1 October 2022 and 29 February 2024. Demographic features, comorbidities, urinary system pathologies, history of urinary tract interventions, hospitalization more than one month prior, antibiotic use within the previous three months, and prior urinary tract infections were compared between patients infected with ESBL-producing and non-ESBL-producing organisms. Antimicrobial susceptibility profiles and the appropriateness of empirical treatments were evaluated. Statistical analyses were performed using SPSS version 25.0, with p < 0.05 considered statistically significant. Results: Escherichia coli (n = 142) and Klebsiella spp. (n = 43) were isolated in 180 of 204 patients. ESBL positivity was detected in 95 patients (52.7%). In the multivariate logistic regression analysis, male sex (p = 0.038) and hospitalization more than one month prior (p = 0.016) were identified as independent risk factors for ESBL positivity, while prior antibiotic use in the last three months showed a borderline association (p = 0.055) and did not reach statistical significance. ESBL production was not associated with prolonged hospitalization; however, bacteremia significantly increased length of stay (p < 0.001). Antimicrobial susceptibility rates were markedly lower in the ESBL-positive group. The appropriateness of empirical therapy was also significantly reduced, with piperacillin–tazobactam being the most frequently inappropriate agent due to high resistance rates and unnecessary broad-spectrum use. Conclusions: ESBL-producing pathogens were highly prevalent among hospitalized patients with pyelonephritis. The low appropriateness of empirical therapy in ESBL-positive cases underscores the need for careful evaluation of ESBL risk factors prior to treatment initiation, as ESBL rates may approach 50%. Full article

(This article belongs to the Special Issue Urinary Tract Infections and Antibiotic Intervention, 2nd Edition)

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12 pages, 340 KB

Open AccessReview

The Angel in the Marble: Antibiotic Duration in the Neonatal Intensive Care Unit

by Joseph B. Cantey and Dalyn B. Guinn

Antibiotics 2026, 15(2), 228; https://doi.org/10.3390/antibiotics15020228 - 20 Feb 2026

Cited by 1 | Viewed by 1559

Abstract

Antimicrobial stewardship in the neonatal intensive care unit is a critically important tool to optimize clinical outcomes. The ideal duration of antimicrobial treatment is a key area that contains many knowledge gaps. This narrative review has three aims. One, to highlight the existing [...] Read more.

Antimicrobial stewardship in the neonatal intensive care unit is a critically important tool to optimize clinical outcomes. The ideal duration of antimicrobial treatment is a key area that contains many knowledge gaps. This narrative review has three aims. One, to highlight the existing evidence for empiric and definitive antibiotic treatment durations for infants; two, to focus on clinical situations where further studies are needed; and three, to propose a rational, goal-based approach to clinical studies that provide for infant safety as shorter treatment durations are investigated. Full article

(This article belongs to the Special Issue Neonatal Infection: Antibiotics for Prevention and Treatment)

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14 pages, 1229 KB

Open AccessArticle

Biofilm Formation in Chicken-Derived Extraintestinal Pathogenic Escherichia coli Alters the Expression of Biofilm- and Virulence-Associated Genes

by Yanze He, Nianling Kuang, Zhihui Chang, Chi Feng, Long Cheng, Jianan Liu, Pei Li, Yuxiang Shi, Fangfang Wang, Yongying Zhang and Cuihong Zhong

Antibiotics 2026, 15(2), 227; https://doi.org/10.3390/antibiotics15020227 - 20 Feb 2026

Viewed by 1098

Abstract

Background: Extraintestinal pathogenic Escherichia coli (ExPEC) poses significant health risks to poultry and humans, with biofilm formation often complicating treatment by enhancing bacterial persistence and resistance. Understanding the genetic mechanisms underlying this lifestyle transition is crucial for controlling infections. This study [...] Read more.

Background: Extraintestinal pathogenic Escherichia coli (ExPEC) poses significant health risks to poultry and humans, with biofilm formation often complicating treatment by enhancing bacterial persistence and resistance. Understanding the genetic mechanisms underlying this lifestyle transition is crucial for controlling infections. This study aimed to investigate the effect of biofilm formation on the transcriptional expression of specific biofilm- and virulence-associated genes in chicken-derived ExPEC strains. Methods: Biofilm formation conditions for three strong biofilm-producing chicken-derived ExPEC strains were optimized using an orthogonal experimental design (L₉(3³)), evaluating culture medium, incubation time, and initial inoculum concentration. Biofilm biomass was quantified via crystal violet staining. Subsequently, the transcription levels of 10 biofilm-associated genes and 17 virulence-associated genes were compared between planktonic and biofilm states using Reverse Transcription-quantitative PCR (RT-qPCR). Results: Optimal culture conditions varied among strains, though nutrient-rich media consistently promoted rapid biofilm formation. Transcriptional analysis revealed significant reprogramming in the biofilm state. Among biofilm-associated genes, flhC, tolA, qseC, mhpB, and bdcR were consistently and significantly upregulated across all strains (p < 0.05). Regarding virulence determinants, the expression of eaeA, LT, fimH, ompF, and iss was significantly upregulated (p < 0.05), whereas Sta levels were significantly reduced (p < 0.05). Conclusions: Biofilm formation induces a distinct transcriptional shift in chicken-derived ExPEC, simultaneously enhancing the expression of key genes involved in biofilm maintenance and pathogenicity. The conserved upregulation of flhC, tolA, qseC, mhpB, and bdcR suggests these genes are critical drivers of biofilm development. Consequently, they represent potential targets for novel therapeutic strategies aimed at preventing E. coli infections and eradicating biofilms in clinical and agricultural settings. Full article

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15 pages, 490 KB

Open AccessReview

Oritavancin for Gram-Positive Bone and Joint Infections: A Comprehensive Review of the Literature

by Zain Ahmed Raza, Alex Giannini and Marco Bongiovanni

Antibiotics 2026, 15(2), 226; https://doi.org/10.3390/antibiotics15020226 - 19 Feb 2026

Cited by 2 | Viewed by 1614

Abstract

Background: Bone and joint infections (BJIs), including osteomyelitis, septic arthritis, and periprosthetic joint infections, typically require prolonged antimicrobial therapy and often involve complex outpatient management. Oritavancin, a long-acting lipoglycopeptide approved for the treatment of acute bacterial skin and skin structure infections caused by [...] Read more.

Background: Bone and joint infections (BJIs), including osteomyelitis, septic arthritis, and periprosthetic joint infections, typically require prolonged antimicrobial therapy and often involve complex outpatient management. Oritavancin, a long-acting lipoglycopeptide approved for the treatment of acute bacterial skin and skin structure infections caused by Gram-positive bacteria, has emerged as a potential off-label option for BJIs owing to its favourable pharmacokinetic and pharmacodynamic properties. Objectives: To provide a comprehensive overview of the pharmacological rationale, microbiological activity, and available clinical evidence supporting the use of oritavancin in BJIs. Methods: A comprehensive narrative review of the literature was performed using MEDLINE and the Cochrane Central Register of Controlled Trials (CENTRAL), focusing on publications from 2011 to 2025. Observational studies, case series, and case reports describing the off-label use of oritavancin in BJIs were considered. Results: The available literature primarily consists of observational studies and real-world experiences. Eighteen studies met the inclusion criteria. Oritavancin was most frequently evaluated for osteomyelitis (n = 14 studies), prosthetic joint infections (n = 10) and septic arthritis (n = 5). Multi-dose regimens, typically including a 1200 mg loading dose followed by weekly doses of 800–1200 mg, were the most commonly described strategies. Reported clinical success rates generally ranged from approximately 70% to over 90%. Oritavancin was overall well tolerated, with adverse events being mostly mild and self-limiting. Conclusions: Current evidence suggests that oritavancin may represent an effective and well-tolerated off-label option for selected patients with Gram-positive BJIs. Its use may offer practical advantages, including reduced hospitalization and avoidance of prolonged intravenous antimicrobial therapy, particularly in patients for whom standard treatment approaches are challenging. Full article

(This article belongs to the Special Issue Diagnostics, Antibiotic Usage and Surgery Related Techniques in Prevention and Treatment of Prosthetic Joint and Implant Related Infections)

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14 pages, 1505 KB

Open AccessArticle

Antibiotic Use Among Children Requiring Respiratory Support in Intensive Care Unit (ICU) from Sofia, Bulgaria: A Single-Center Retrospective Experience

by Lilia Bozadzhieva, Dimitrinka Miteva, Lyubomila Ilarionova, Tania Teneva, Blagomir Zdravkov and Guergana Petrova

Antibiotics 2026, 15(2), 225; https://doi.org/10.3390/antibiotics15020225 - 19 Feb 2026

Cited by 1 | Viewed by 1137

Abstract

Antibiotic use in critically ill children requiring respiratory support remains controversial, particularly in the absence of standardized guidelines for patients managed with non-invasive ventilation (NIV). Evidence in this area remains limited, and real-world data are therefore valuable. Objective: This retrospective single-center study aimed [...] Read more.

Antibiotic use in critically ill children requiring respiratory support remains controversial, particularly in the absence of standardized guidelines for patients managed with non-invasive ventilation (NIV). Evidence in this area remains limited, and real-world data are therefore valuable. Objective: This retrospective single-center study aimed to describe antibiotic prescribing patterns and infectious outcomes in pediatric patients admitted to the intensive care unit (PICU) with respiratory failure, according to the type of respiratory support. Methods: Children aged 0–17 years admitted between January 2021 and February 2025 who required oxygen supplementation, NIV, or invasive mechanical ventilation (IMV) were included. Demographic characteristics, underlying conditions, infectious complications, antibiotic exposure, length of PICU stay, and outcomes were analyzed using descriptive statistics and univariate comparisons. Results: Eighty-nine patients were included. Ventilator-associated pneumonia (VAP) occurred exclusively in patients receiving IMV, and infection complications were observed more in this group compared to those receiving NIV (p = 0.005). Pseudomonas aeruginosa was the most frequently isolated pathogen. Antibiotics were administered in 82% of patients, with no significant association between the respiratory support and initiation of antibiotic therapy (p = 0.195). A higher number of antibiotics was administered in patients receiving IMV compared with those receiving oxygen therapy alone. Conclusions: Antibiotic use in children requiring respiratory support in the PICU was common and appears to be driven primarily by underlying disease and illness severity rather than by the ventilation modality alone. Infections specific to invasive ventilation, such as VAP, were more frequent in patients receiving IMV, while infection-related outcomes in non-invasive groups should be interpreted cautiously due to differences in diagnostic definitions. These findings are descriptive and hypothesis-generating and highlight the need for prospective multicenter studies to create evidence-based antibiotic stewardship strategies in pediatric critical care. Full article

(This article belongs to the Special Issue Therapy of Infectious Diseases Among Children and Adults: The Role of Antibiotics in Daily Practice)

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25 pages, 7494 KB

Open AccessArticle

Escaping the ESKAPE Antimicrobial Resistant Cycle with EVQ-218

by Ali Sakawa Sharif, Kayla C. Maas, Isabella Fratangelo, Kenneth J. Woolley, David B. Nilson and William H. Niedermeyer

Antibiotics 2026, 15(2), 224; https://doi.org/10.3390/antibiotics15020224 - 18 Feb 2026

Viewed by 2103

Abstract

Background/Objectives: Antimicrobial resistance (AMR) continues to expand under sustained exposure to conventional antibiotics, contributing to the emergence of multidrug- and pan-resistant bacterial pathogens. There remains a critical need for antimicrobial agents that maintain activity during prolonged selective pressure while minimizing the potential for [...] Read more.

Background/Objectives: Antimicrobial resistance (AMR) continues to expand under sustained exposure to conventional antibiotics, contributing to the emergence of multidrug- and pan-resistant bacterial pathogens. There remains a critical need for antimicrobial agents that maintain activity during prolonged selective pressure while minimizing the potential for resistance development. This study aimed to evaluate EVQ-218, a non-ionic silver-based antimicrobial, against World Health Organization-designated ESKAPE pathogens. Methods: EVQ-218 was assessed using extended serial passage experiments performed under both sub- and supra-minimum inhibitory concentration (MIC) conditions. Comparative resistance selection experiments were conducted in parallel using tobramycin and ciprofloxacin, and susceptibility was evaluated through MIC determination and phenotypic analysis. Results: Across extended serial passage experiments, EVQ-218 did not exhibit measurable increases in MIC or phenotypic indicators of adaptive resistance. In contrast, parallel exposure to tobramycin and ciprofloxacin resulted in rapid and sustained MIC elevation. Notably, isolates that acquired resistance to either comparator antibiotic retained susceptibility to EVQ-218, indicating a lack of cross-resistance. Mechanistic analyses were consistent with a non-lytic, intracellular mode of antibacterial activity involving disruption of sulfur-associated biomolecular processes, suggestive of a multi-site mechanism distinct from classical antibiotics. Conclusions: These findings support EVQ-218 as a promising broad-spectrum antimicrobial candidate with resistance-resilient activity and warrant further investigation of its potential role in addressing unmet needs in AMR therapeutics. Full article

(This article belongs to the Special Issue Innovations in Antibacterial Agents: Combating Resistance, Improving Efficacy and Exploring New Mechanisms)

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19 pages, 2748 KB

Open AccessArticle

Penicillin G and Cloxacillin in Costa Rican Dairy Products: Quantitative Analysis and Lactic Acid Bacteria Resistance Profiling

by María Cristina Solís-Robles, Melissa Quesada-Solano, Fabio Granados-Chinchilla, Carolina Cortés-Herrera, Mauricio Redondo-Solano and Adriana Fernández-Campos

Antibiotics 2026, 15(2), 223; https://doi.org/10.3390/antibiotics15020223 - 18 Feb 2026

Cited by 1 | Viewed by 1954

Abstract

Background/Objectives: Milk and dairy products are among the most relevant foods both nutritionally and commercially. Costa Rica stands out as one of the main producers and consumers of dairy products in Central America. However, in recent years, the use of antibiotics in the [...] Read more.

Background/Objectives: Milk and dairy products are among the most relevant foods both nutritionally and commercially. Costa Rica stands out as one of the main producers and consumers of dairy products in Central America. However, in recent years, the use of antibiotics in the livestock industry has increased, with implications for public health and food security, generating a need to monitor residues of these drugs in food. The present research focuses on developing a liquid chromatography method for the simultaneous quantification of penicillin G (PEN) and cloxacillin (CLO) in raw and commercial bovine milk, as well as in various dairy products, including fresh cheese and liquid yogurt. Methods/Results: During the validation of the methodology, average sensitivities of (960 ± 8)·10¹ mg L⁻¹ and (1580 ± 9)·10¹ mg L⁻¹ were achieved for PEN and CLO, respectively. Determination coefficients of 0.9995 and 0.9998 were also achieved, respectively. The limits of detection (LOD) and quantification (LOQ) for PEN and CLO were (0.330 ± 0.025) mg L⁻¹ and (0.65 ± 0.12) mg L⁻¹, (1.10 ± 0.083) mg L⁻¹ and (2.2 ± 0.4) mg L⁻¹, respectively, on both accounts. Recoveries were 68–77%, 92–106%, and 78–112% for PEN and 57–79%, 99–114%, and 95–120% for CLO in milk, cheese, and yogurt, respectively, across all three concentration levels evaluated. The precision of the method, in terms of reproducibility, was assessed for liquid yogurt (3–12% RSD_R for PEN and 4–12% RSD_R for CLO) and in cheese (8–14% RSD_R for PEN and 4–12% RSD_R for CLO). Nineteen bovine milk samples from the Cartago area were evaluated, including commercial and milk samples. Additionally, cheese (n = 17) and yogurt samples (n = 8) were analyzed. Conclusions: None of the samples showed detectable signals of the antibiotics. In addition, antibiotic sensitivity testing was performed on n = 9 Lactic Acid Bacteria (LAB) strains isolated from the raw milk samples, revealing the presence of some resistant traits to several antibiotics, including β-lactams. Full article

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20 pages, 632 KB

Open AccessArticle

Aminoglycoside Resistance Among Clinical Bacterial Isolates in Sétif, Algeria: Epidemiology, Multidrug Resistance, and Virulence Features

by Anfal Kara, Chiara Massaro, Naouel Boussoualim, Giovanni M. Giammanco, Rosa Alduina, Zineb Daoudi, Noussaiba Douadi, Fatma Gridi, Mohammad Raish, Byong-Hun Jeon, Hyun-Jo Ahn and Yacine Benguerba

Antibiotics 2026, 15(2), 222; https://doi.org/10.3390/antibiotics15020222 - 17 Feb 2026

Viewed by 1313

Abstract

Background. Antibiotic resistance is a growing global health challenge, complicating the management of infections. Aminoglycosides are increasingly associated with resistance, raising the risk of clinical complications and mortality in severe infections. This study aimed to characterize the epidemiological profile of 135 aminoglycoside-resistant [...] Read more.

Background. Antibiotic resistance is a growing global health challenge, complicating the management of infections. Aminoglycosides are increasingly associated with resistance, raising the risk of clinical complications and mortality in severe infections. This study aimed to characterize the epidemiological profile of 135 aminoglycoside-resistant clinical strains collected in Setif between 2021 and 2023. Methods. Antibiotic susceptibility testing was performed according to EUCAST guidelines, and phenotypic assays were conducted to assess key virulence traits, including biofilm formation and enzyme production. Results. Aminoglycoside resistance was more frequently observed in female patients (55.6%) and was found to be predominant among adults (68.1%). Urinary tract infections represented the main clinical presentation (76.3%), with Escherichia coli being the most common isolate (40.7%). High resistance rates were detected for amoxicillin (83%), amoxicillin–clavulanic acid (80%), cephalexin (74.8%), cefixime (71.1%), trimethoprim–sulfamethoxazole (74.8%), and gentamicin (72.6%). Conversely, chloramphenicol (53.3%), imipenem (47.4%), amikacin (47.4%), and piperacillin–tazobactam (31.1%) remained comparatively more effective. Multidrug resistance involving seven antibiotics occurred in 25.6% of isolates, with notable cross-resistance patterns between gentamicin and β-lactam antibiotics (5 out of 22). Genotypic analysis showed that 43% of isolates carried at least one β-lactamase gene, whereas 9.6% harbored a qnr determinant. Regarding virulence factors, isolates with low biofilm-forming ability were found to be the most common (62.96%). Conclusion. In conclusion, this study revealed substantial variations in aminoglycoside resistance in Setif, shaped by demographic, clinical, and bacteriological factors. Full article

(This article belongs to the Section Mechanism and Evolution of Antibiotic Resistance)

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27 pages, 953 KB

Open AccessArticle

Arms-Based Meta-Analysis of Microbiological Endpoints of 88 VAP Prevention Studies Using Antimicrobial Versus Non-Antimicrobial Strategies—Towards ‘VAP-Zero’?

by James C. Hurley

Antibiotics 2026, 15(2), 221; https://doi.org/10.3390/antibiotics15020221 - 17 Feb 2026

Viewed by 971

Abstract

Background/Objectives: In traditional contrast-based meta-analyses of randomized concurrent controlled trials (RCCTs), topical antibiotic prophylaxis (TAP) appears more effective than either antiseptic-based or non-antimicrobial-based interventions for preventing ventilator-associated pneumonia (VAP). The objective here is to use arm-based methods to determine whether this effectiveness translates [...] Read more.

Background/Objectives: In traditional contrast-based meta-analyses of randomized concurrent controlled trials (RCCTs), topical antibiotic prophylaxis (TAP) appears more effective than either antiseptic-based or non-antimicrobial-based interventions for preventing ventilator-associated pneumonia (VAP). The objective here is to use arm-based methods to determine whether this effectiveness translates towards achieving VAP-zero, both overall and specifically for VAP in association with Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter species among the same RCCTs. Methods: Data were extracted from RCCTs sourced primarily from Cochrane reviews of VAP prevention interventions. Arms-based and contrast-based methods of meta-analyses of the VAP prevention effect size and the VAP incidence per 100 patients receiving mechanical ventilation were obtained using random effects methods. Results: The VAP prevention intervention effect sizes derived by contrast-based versus arms-based meta-analyses were similar for each of the three broad types of interventions. The overall VAP prevention effect of antibiotic-based interventions by contrast-based and arms-based methods were 0.39 (95% confidence interval 0.33 to 0.46; n = 28) versus 0.39 (95% confidence interval 0.32 to 0.47; n = 28), respectively. Surprisingly, the arms-based analysis revealed that the summary VAP incidence, both overall and for each of Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter species within antibiotic intervention groups, were similar to the respective summary incidences within intervention groups of non-antimicrobial RCCTs. Conclusions: VAP-zero, both overall and in association with specific microbial sub-types, has remained elusive using antimicrobial-based interventions. This inference was not evident from a contrast-based analysis. Full article

(This article belongs to the Special Issue Antimicrobial Stewardship and Infection Prevention in Intensive Care Unit)

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17 pages, 1700 KB

Open AccessEditor’s ChoiceArticle

Risk Factors Associated with the Emergence of Multidrug-Resistant Bacteria and Fungal Infections in Walled-Off Pancreatic Necrosis

by Michael Fernandez Y Viesca, Alia Hadefi, Lukas Otero Sanchez, Martina Pezzullo, Morgane Van Wettere, Eleni Karakike, Maya Hites, Viviane De Maertelaer, Myriam Delhaye and Marianna Arvanitakis

Antibiotics 2026, 15(2), 220; https://doi.org/10.3390/antibiotics15020220 - 17 Feb 2026

Cited by 1 | Viewed by 1400

Abstract

Background: Infected pancreatic necrosis (IPN) is a serious complication of moderate-to-severe acute pancreatitis (AP), associated with high morbidity, intensive care unit (ICU) admission, organ failure, and mortality. Initial management relies on antibiotics and drainage of walled-off necrosis (WON). In the context of increasing [...] Read more.

Background: Infected pancreatic necrosis (IPN) is a serious complication of moderate-to-severe acute pancreatitis (AP), associated with high morbidity, intensive care unit (ICU) admission, organ failure, and mortality. Initial management relies on antibiotics and drainage of walled-off necrosis (WON). In the context of increasing multidrug-resistant (MDR) bacteria, identifying risk factors for MDR emergence is crucial. The impact of fungal infections (FIs) on outcomes also remains unclear. This study aimed to identify risk factors associated with the emergence of MDR bacteria and FIs during intervention for IPN. Methods: This retrospective study included 71 consecutive patients undergoing intervention for suspected IPN or symptomatic WON. Results: At first intervention, IPN was confirmed in 52 patients (73%), MDR bacteria in 19 (27%), extensively drug-resistant (XDR) bacteria in 4 (5.6%), and FI in 21 (30%). After all interventions, MDR/XDR bacteria and fungi were detected in 25 (35%)/11 (15.5%) and 42 (59%) patients, respectively. Independent risk factors for MDR emergence were the number of antibiotic changes (b, 1.70; 95% CI 1.18–2.43; p = 0.004) and need for nutritional support (NS) (b, 5.69; 95% CI 1.52–20.50; p = 0.010). No independent factor was associated with FI. The 180-day mortality did not differ across groups. The 90-day cumulative ICU admission rate was higher in IPN vs. non-IPN (63.1% vs. 29.4%, p = 0.030) and in MDR vs. non-MDR (72.2% vs. 37.1%, p = 0.005). Conclusions: Antibiotic changes and NS were independently associated with MDR emergence in IPN. No independent factors were linked to FI. ICU admission was significantly higher in IPN and MDR cases. Full article

(This article belongs to the Special Issue Therapy of Infectious Diseases Among Children and Adults: The Role of Antibiotics in Daily Practice)

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20 pages, 4280 KB

Open AccessArticle

Silybin Interferes with the Intracellular Replication of Piscirickettsia salmonis in SHK-1 Cells and Confers Protection in Salmo salar

by Mick Parra, Meraiot Rubio, Katherin Izquierdo, Valentina Barsotti, Ana María Sandino and Brenda Modak

Antibiotics 2026, 15(2), 219; https://doi.org/10.3390/antibiotics15020219 - 17 Feb 2026

Cited by 1 | Viewed by 954

Abstract

Background/Objectives: The salmon industry plays an important role in the Chilean economy, positioning the country as the second-largest producer of salmonids worldwide after Norway. However, this rapid growth has led to an increase in outbreaks of infectious diseases, which cause significant economic [...] Read more.

Background/Objectives: The salmon industry plays an important role in the Chilean economy, positioning the country as the second-largest producer of salmonids worldwide after Norway. However, this rapid growth has led to an increase in outbreaks of infectious diseases, which cause significant economic losses to the industry. The pathogen that most affects the salmon industry is the bacterium Piscirickettsia salmonis, accounting for 43.1% of infection-related deaths. In the search for new treatment alternatives against P. salmonis, we have previously reported that the effect of co-incubating silybin at sub-IC50 concentrations decreases the intracellular presence of P. salmonis in SHK-1 cells. Methods: This article evaluates the effect of silybin on the immune response and oxidative stress of SHK-1 cells infected with P. salmonis, as well as the reduction in intracellular bacterial replication during the first 72 h of infection. Furthermore, we assess the ability of silybin administration to modulate the immune response in S. salar and protect against P. salmonis infection. Results: The results show that co-incubation of silybin during infection in SHK-1 cells modulates the expression levels of the genes gsh-px, cat, tnf-α, and il-1β and also decreases the levels of intracellular ROS generated by the infection. Furthermore, the mechanism of action of silybin in SHK-1 cells is related to interference with the intracellular replication of P. salmonis after 72 h of infection and not to adherence or internalization of the bacteria. Finally, silybin is able to generate protection in S. salar infected with P. salmonis independently of stimulation of the immune response. Conclusions: In conclusion, silybin administration may be an effective treatment against P. salmonis in salmonids; however, further studies are needed to clarify the mechanism of action. Full article

(This article belongs to the Special Issue Antibiotics and Antimicrobial Resistance in Aquaculture: Alternative and Complementary Treatment Methods)

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18 pages, 1501 KB

Open AccessArticle

Imipenem in the Rat Brain: A Multidimensional Study on Hippocampal Behavior, GABAergic System, Astrocyte Response, and Neurogenesis

by Leonardo Araújo-Andrade, Bárbara Caetano-Mota, Inês Silva, Ana Rogeiro, Pedro Nogueira, Ana Silva, Pedro A. Pereira, Maria Dulce Madeira and Armando Cardoso

Antibiotics 2026, 15(2), 218; https://doi.org/10.3390/antibiotics15020218 - 17 Feb 2026

Cited by 1 | Viewed by 1150

Abstract

Background: After imipenem was introduced in clinical practice, neurologic adverse effects led to recommendations against its use in patients with neurologic conditions. However, these conclusions were drawn without considering pharmacokinetic variations in such patients. Furthermore, animal studies lack the use of clinically relevant [...] Read more.

Background: After imipenem was introduced in clinical practice, neurologic adverse effects led to recommendations against its use in patients with neurologic conditions. However, these conclusions were drawn without considering pharmacokinetic variations in such patients. Furthermore, animal studies lack the use of clinically relevant doses and supporting morphological studies in both naïve and disease models. Objectives: We aim to study the effects of imipenem in the hippocampus of naïve animals, evaluating potential behavioral and morphological alterations. Methods: Naïve Wistar rats received a 10-day course of intraperitoneal imipenem (40 mg/kg) while controls received a saline injection. After that, they were put through the Morris water maze, elevated plus maze, open-field test, and then euthanized. We analyzed neurogenesis (using doublecortin immunoreactivity), astrogliosis, and the γ-Aminobutyric acid (GABA)ergic system (using parvalbumin (PV), calretinin (CR) and calbindin (CB) immunoreactive (IR) neurons) in the hippocampus. Results: Interestingly, our results showed no significant alterations in both short and long-term memory, nor in anxiety. There were also no significant changes in the neuronal density of doublecortin-immunoreactive (IR) neurons nor in astrogliosis. Furthermore, the areal density of PV- and CR-IR was preserved in all hippocampal subfields. The density of CB-IR neurons also showed no changes in the dentate gyrus, CA3, and subiculum; however, a significant increase was found in the CA1 region. Conclusions: Our results indicate that in naïve individuals, a clinically relevant dose of imipenem does not seem to cause overt behavioral deficits or widespread morphological alterations in the hippocampus. However, a specific increase in the CB-IR neuronal population in the CA1 region highlights a localized cellular alteration/plasticity induced by the imipenem. Hence, pharmacokinetic factors seem to be the potential contributors of imipenem side effects. Further studies should focus on this as a possible cause and focus on individuals with brain diseases. Full article

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18 pages, 929 KB

Open AccessArticle

Susceptibility Profile and Multiple Antibiotics Resistance of Escherichia coli, Klebsiella spp., and Enterococci from Small-Scale Cattle Farms in Tennessee

by Goodness Olakanmi, Maureen Nzomo, Bharat Pokharel, Abdullah Mafiz and Agnes Kilonzo-Nthenge

Antibiotics 2026, 15(2), 217; https://doi.org/10.3390/antibiotics15020217 - 17 Feb 2026

Viewed by 1260

Abstract

Background/Objectives: Antimicrobial resistance in food–animal environments threatens sustainable production and public health, yet small farms remain poorly characterized as potential reservoirs of antimicrobial resistant bacteria. To address this, we investigated the prevalence and antimicrobial resistance profiles of Escherichia coli, Klebsiella spp., [...] Read more.

Background/Objectives: Antimicrobial resistance in food–animal environments threatens sustainable production and public health, yet small farms remain poorly characterized as potential reservoirs of antimicrobial resistant bacteria. To address this, we investigated the prevalence and antimicrobial resistance profiles of Escherichia coli, Klebsiella spp., and Enterococcus spp. from small-scale cattle farms in Tennessee, USA. Methods: Over one year, 153 environmental samples (soil, manure, water) were collected from 17 farms. Target bacteria were isolated and confirmed using selective agar, biochemical tests, and PCR, and tested against 12 antibiotics using the Kirby–Bauer disk diffusion test. Multiple Antibiotic Resistance Index (MARI) and multidrug resistance (MDR) profiles were summarized. A complementary farmer survey of 26 farmers captured veterinary access, antibiotic use, manure handling, record keeping, and awareness of antimicrobial resistance. Results: Prevalence was highest for Enterococcus spp. (41.8%), followed by E. coli (23.5%) and Klebsiella spp. (12.4%). Seasonal variation was significant for E. coli and Enterococcus (p < 0.05). Winter manure yielded highest detection of E. coli (55.6%) and Enterococcus (53.8%), whereas Klebsiella peaked in Fall soil (19.1%). Resistance patterns varied across species, with Enterococcus showing consistent resistance to all three. E. coli frequently resisted erythromycin, ampicillin, and azithromycin; and Klebsiella commonly resisted erythromycin, ampicillin, and cefotaxime, though some of these reflect intrinsic resistance rather than acquired clinical resistance. MARI values were 0.92 in manure and soil, identifying them as high-risk reservoirs. We identified 29 distinct MDR pattern. Bipartite network visualization highlighted “resistance hubs” around erythromycin, ampicillin, and vancomycin, particularly in Enterococcus. In our study, 76.9% of farmers consulted veterinarians before antibiotic use, 57.7% kept written antibiotic records, and 65.4% were aware of AMR as a public health issue. Small-scale cattle farms are potential reservoirs of multidrug resistant commensal bacteria. Conclusions: These findings provide an evidence-based foundation to guide targeted antimicrobial stewardship and promote sustainable management practices in small-scale food animal farms. Full article

(This article belongs to the Section Antibiotics in Animal Health)

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12 pages, 207 KB

Open AccessFeature PaperArticle

Implementing Information Resources to Support Shared Decisions in Australian Primary Care: A Qualitative Perspective of an Antimicrobial Stewardship Strategy

by Ruby Biezen, Kaleswari Somasundaram, Stephen Ciavarella, Tim Monaghan, Kirsty Buising and Jo-Anne Manski-Nankervis

Antibiotics 2026, 15(2), 216; https://doi.org/10.3390/antibiotics15020216 - 17 Feb 2026

Viewed by 1106

Abstract

Background/Objectives: Inappropriate use of antibiotics contributes to the rise in antibiotic-resistant bacteria and can result in adverse drug effects for individuals. Informed discussions between patients and general practitioners (GPs) can help ensure that treatment decisions about antibiotic use align with the best health [...] Read more.

Background/Objectives: Inappropriate use of antibiotics contributes to the rise in antibiotic-resistant bacteria and can result in adverse drug effects for individuals. Informed discussions between patients and general practitioners (GPs) can help ensure that treatment decisions about antibiotic use align with the best health outcomes for individuals. Methods: We implemented a set of information resources designed to support clinical discussions and decision-making for patients with common infections in primary care. A suite of patient information sheets, which had been co-designed with primary care providers and consumers, were implemented in eight general practices in metropolitan Melbourne and regional Victoria, from August to November 2020. Results: Post-implementation evaluation, conducted through interviews with 15 primary care providers and 13 patients, revealed that the information sheets were simple, easy to use and generated discussion within consultations. GPs reported using the sheets to reinforce their decision-making during consultations with patients, reduce potential conflict, provide alternatives to antibiotic prescriptions, and offer patients a written summary of management recommendations. Patients found the sheets informative and that they made it easier to understand their diagnosis and to manage their conditions. Both GPs and patients agreed that the content was relevant and effectively enhanced patients’ knowledge of disease conditions, treatment options, and when to seek medical advice and were facilitators of meaningful conversations during consultations. Conclusion: These resources are acceptable in Australian primary care and publicly available for use by GPs, pharmacists and patients in Australia. Full article

(This article belongs to the Special Issue Managing Appropriate Antibiotic Prescribing and Use in Primary Care)

13 pages, 2895 KB

Open AccessArticle

Evaluation of the Efficacy of Anthelmintic Drugs Against Trichinella spiralis Larvae

by Soon-Ok Lee, Su In Heo, Hyeon-Woo Nam, Ji-Hyun Lee, Ki Back Chu, Gi-Ja Lee, Tong In Oh, Sung Soo Kim and Fu-Shi Quan

Antibiotics 2026, 15(2), 215; https://doi.org/10.3390/antibiotics15020215 - 16 Feb 2026

Viewed by 1474

Abstract

Background: Albendazole, mebendazole, and ivermectin are effective against adult Trichinella spiralis but show limited efficacy against encapsulated muscle stage larvae. This limitation highlights the need for improved experimental approaches to evaluate anthelmintic activity at this stage and to identify alternative therapeutic candidates. [...] Read more.

Background: Albendazole, mebendazole, and ivermectin are effective against adult Trichinella spiralis but show limited efficacy against encapsulated muscle stage larvae. This limitation highlights the need for improved experimental approaches to evaluate anthelmintic activity at this stage and to identify alternative therapeutic candidates. Methods: Seven antiparasitic drugs, albendazole (ABZ), miltefosine (MLT), ivermectin (IVM), tribendimidine (TBD), praziquantel (PZQ), artesunate (ART), and mefloquine (MEQ), were evaluated for in vitro activity against T. spiralis muscle larvae. Larval viability was quantified using a tetrazolium salt XTT assay to determine IC₅₀ values and compare with microscopic assessments. Based on in vitro activity, TBD was selected for in vivo evaluation in a mouse model, where efficacy was assessed by muscle larval burden and histopathological changes. Results: TBD, MEQ, IVM, and ABZ exhibited measurable in vitro efficacies against T. spiralis larvae, with TBD showing the lowest IC₅₀ value at 135.2 μM. XTT formazan absorbance correlated strongly with larval number and incubation time. In vivo, TBD treatment significantly reduced larval burdens in diaphragm and gastrocnemius muscles and was associated with reduced collagen capsule thickness, inflammation, and fibrosis compared with ABZ-treated controls. Conclusions: This study validated an assay for quantitative evaluation of T. spiralis muscle larvae and demonstrates robust in vitro and in vivo activity of TBD against this stage. Full article

(This article belongs to the Section Novel Antimicrobial Agents)

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22 pages, 1234 KB

Open AccessArticle

Etiologic Patterns and Evolution of Healthcare-Associated Infections in the Pandemic and Post-Pandemic Periods: A County-Level Multicenter Study from Southeastern Romania

by Corina Voinea, Elena Mocanu, Elena Dantes, Sanda Jurja, Ana-Maria Neculai, Aurora Craciun, Lucian Serbanescu, Ana-Maria Dascalu, Mihaela Cezarina Mehedinti and Sorin Rugina

Antibiotics 2026, 15(2), 214; https://doi.org/10.3390/antibiotics15020214 - 15 Feb 2026

Viewed by 826

Abstract

Background/Objectives: Healthcare-associated infections (HAIs) remain a major source of morbidity, mortality, and healthcare burden, and were profoundly affected by the COVID-19 pandemic through changes in case mix, care organization, and antimicrobial use. This study aimed to compare the epidemiology, etiology, ward distribution, [...] Read more.

Background/Objectives: Healthcare-associated infections (HAIs) remain a major source of morbidity, mortality, and healthcare burden, and were profoundly affected by the COVID-19 pandemic through changes in case mix, care organization, and antimicrobial use. This study aimed to compare the epidemiology, etiology, ward distribution, risk factors, and outcomes of HAIs during the pandemic and post-pandemic periods in southeastern Romania, with particular emphasis on Clostridioides difficile infection (CDI), multidrug-resistant (MDR) pathogens, and in-hospital mortality. Methods: This retrospective observational study included 3929 patients with confirmed HAIs reported by 10 hospitals in one Romanian county between March 2020 and December 2024, divided into a pandemic period (March 2020–March 2022) and a post-pandemic period (April 2022–December 2024). Sociodemographic, clinical, ward-related, therapeutic, and microbiological variables, together with discharge status and cause of death, were analyzed using Fisher’s exact test, Z-tests with Bonferroni correction, the Mann–Whitney U test, and multivariable models, applying national and ECDC-aligned surveillance definitions for HAIs. Results: Patients were predominantly older adults (median age 67 years), with a slight male and urban predominance. Hospital stays were longer during the pandemic. Immunosuppression, previous surgery, antisecretory therapy, and chemotherapy were more frequent post-pandemic. HAIs were mainly reported from medical wards, with a relative shift towards intensive care units during the pandemic; pediatric wards carried a smaller burden. CDI was the leading HAI (about half of all cases) with higher post-pandemic prevalence, whereas SARS-CoV-2 infections predominated in medical and surgical wards; Acinetobacter baumannii and Klebsiella pneumoniae clustered in intensive care units during the pandemic, and were more often associated with mortality. Overall, 59.7% of patients improved and 17.5% died, with higher mortality during the pandemic, while post-pandemic deaths were more frequently unrelated to HAIs. Conclusions: This study demonstrates a substantial and ongoing burden of healthcare-associated infections in southeastern Romania, with elderly patients with prolonged hospital stays and complex medical conditions being most affected and experiencing considerable mortality, particularly in medical and intensive care units. After the pandemic, Clostridioides difficile infections became more prevalent in the context of repeated antibiotic use and immunosuppression. Mortality among patients with HAIs was higher during the pandemic, whereas in the post-pandemic period deaths were more often unrelated to HAIs, underscoring the need to strengthen antimicrobial stewardship programs and infection prevention strategies. Full article

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15 pages, 2552 KB

Open AccessArticle

A Cyclic Pentapeptide Inhibits AgrC as a Quorum-Sensing Quenching Agent in Staphylococcus aureus

by Duiyuan Ai, Huanhuan Duan and Jiahao Yao

Antibiotics 2026, 15(2), 213; https://doi.org/10.3390/antibiotics15020213 - 15 Feb 2026

Cited by 1 | Viewed by 938

Abstract

Background/Objectives: Staphylococcus aureus virulence is tightly regulated by the agr (accessory gene regulator) quorum-sensing system. Targeting AgrC, the histidine kinase receptor that serves as a core regulator of agr signaling, represents a promising antivirulence strategy that circumvents conventional bactericidal pressure. Methods: In this [...] Read more.

Background/Objectives: Staphylococcus aureus virulence is tightly regulated by the agr (accessory gene regulator) quorum-sensing system. Targeting AgrC, the histidine kinase receptor that serves as a core regulator of agr signaling, represents a promising antivirulence strategy that circumvents conventional bactericidal pressure. Methods: In this study, structure-based virtual screening using AutoDock Vina was performed, followed by molecular dynamics simulations, to identify potent analogs of known AgrC inhibitors. Results: A cyclo[Ala-Phe-OLeu-Phe-D-Leu] exhibiting high binding affinity and stable receptor interaction was selected for further evaluation. Antimicrobial susceptibility testing confirmed that the compound did not inhibit bacterial growth. However, at a concentration of 16 µg/mL, it significantly inhibited hemolytic activity with high reproducibility, and the inhibition rate reached 77.60%. Quantitative reverse transcription PCR (RT-qPCR) demonstrated that the compound decreased some key AgrC-mediated genes, including agrC, agrA, saeS, hla, spa, fnbA, and lukS. Conclusions: These findings identify a promising cyclic pentapeptide inhibitor of AgrC that effectively attenuates S. aureus virulence without exerting bactericidal pressure. This work provides a valuable lead compound and offers novel insights for the development of advanced, safe, and effective antivirulence therapeutics. Full article

(This article belongs to the Section Novel Antimicrobial Agents)

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27 pages, 14586 KB

Open AccessArticle

Genomic Characterisation of Pyometra-Associated Escherichia coli in a Lombardy Veterinary Clinic: A Nanopore-Based Case Series

by Gabriele Meroni, Alessio Soggiu, Davide Sciannimanico, Raul Alexandru Pop, Luigi Bonizzi and Piera Anna Martino

Antibiotics 2026, 15(2), 212; https://doi.org/10.3390/antibiotics15020212 - 15 Feb 2026

Viewed by 2264

Abstract

Background/Objectives: Pyometra is a life-threatening uterine infection of intact bitches and queens. Despite growing reports of multidrug-resistant (MDR) Escherichia coli in canine reproductive and urinary infections, no whole-genome data were previously available for pyometra isolates from Italy. This study aimed to characterise, by [...] Read more.

Background/Objectives: Pyometra is a life-threatening uterine infection of intact bitches and queens. Despite growing reports of multidrug-resistant (MDR) Escherichia coli in canine reproductive and urinary infections, no whole-genome data were previously available for pyometra isolates from Italy. This study aimed to characterise, by whole-genome sequencing and comparative genomics, the population structure, resistome and virulome of E. coli causing pyometra in companion animals from northern Italy in the context of European datasets. Methods: Four E. coli isolates (two canine, two feline) from pyometra cases underwent nanopore long-read sequencing. Genomes were compared with Brazilian and Finnish pyometra isolates using core- and accessory-genome analyses, pan-genome partitioning, phylogeny, and gene-based profiling of antimicrobial resistance and virulence determinants. Results: All Italian isolates belonged to phylogroup B2 and to recognised ExPEC sequence types (ST706/O51:H1, ST141/O2:H6, ST372/O75:H31, ST646/O22:H5). Phenotypically, they were uniformly resistant to several penicillins and early/third-generation cephalosporins but remained susceptible to fluoroquinolones, aminoglycosides and trimethoprim–sulphonamide. The combined 57-genome pan-genome was open yet strongly core-dominated; Italian strains shared an efflux- and regulator-centred intrinsic resistome and a rich ExPEC virulence repertoire (P, S, F1C and type 1 fimbriae, multiple siderophores, colibactin, Vat, haemolysin, CNF1) with Brazilian and Finnish isolates. Conclusions: Pyometra-associated E. coli from northern Italian pets belong to globally disseminated high-risk B2 lineages that combine extensive virulence with a largely intrinsic resistome, and currently retain susceptibility to several key drug classes, underscoring an important but vulnerable therapeutic window. Full article

(This article belongs to the Special Issue Gram-Negative Infections in Humans and Companion Animals: A Global Threat)

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12 pages, 2335 KB

Open AccessCommunication

The Antimicrobial Peptide C14R Is Active Against All Pathogenic Species of the ESKAPE Group

by Daniel Gruber, Verena Vogel, Jan-Christoph Walter, Grigory Bolotnikov, Armando Rodríguez, Nico Preising, Ludger Ständker, Carolina Firacative, Barbara Spellerberg, Ann-Kathrin Kissmann and Frank Rosenau

Antibiotics 2026, 15(2), 211; https://doi.org/10.3390/antibiotics15020211 - 15 Feb 2026

Cited by 1 | Viewed by 1258

Abstract

The global rise in antimicrobial resistance among the ESKAPE pathogens represents a major challenge to public health. Here, we report the broad-spectrum antibacterial activity of the synthetic antimicrobial and pore-forming peptide C14R against all six ESKAPE species. Using a radial diffusion assay and [...] Read more.

The global rise in antimicrobial resistance among the ESKAPE pathogens represents a major challenge to public health. Here, we report the broad-spectrum antibacterial activity of the synthetic antimicrobial and pore-forming peptide C14R against all six ESKAPE species. Using a radial diffusion assay and resazurin-based viability testing, C14R exhibited a potent bactericidal effect with minimum inhibitory concentrations (MICs), defined as the lowest concentration of an antimicrobial agent that completely inhibits visible growth of planktonic microorganisms, ranging from 3.4 µg/mL (Enterococcus faecium, vancomycin-resistant) to 45.2 µg/mL (Klebsiella quasipneumoniae, ESBL). C14R also inhibited biofilm formation by Gram-positive pathogens, with minimum biofilm inhibitory concentrations (MBICs), referring to the minimal concentration required to prevent the development of biofilms, of 15.0 µg/mL (Staphylococcus aureus, MRSA) and 22.0 µg/mL (E. faecium, VRE), whereas Gram-negative biofilms showed higher tolerance. Together, these findings demonstrate that C14R retains high activity against multidrug-resistant ESKAPE strains, highlighting its potential as a lead compound for the development of next-generation antimicrobial drugs to expand the portfolio of available antibiotics and brace health systems against emerging severe infections. Full article

(This article belongs to the Special Issue Antimicrobial Peptides (AMPs) Against Human Pathogens)

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16 pages, 1057 KB

Open AccessReview

From Bench to Bedside: Personalized Genomics in the Diagnosis and Treatment of Osteomyelitis

by Amir Human Hoveidaei, Arian Rahimzadeh, Sara Mohammadi, Pranav Thota, Kimia Vakili, Parsa Yazdanpanahi, Ali Homaei, Seyed Arad Mosalamiaghili, Jakob Adolf and Janet D. Conway

Antibiotics 2026, 15(2), 210; https://doi.org/10.3390/antibiotics15020210 - 14 Feb 2026

Cited by 1 | Viewed by 1424

Abstract

Osteomyelitis (OM), an inflammatory condition of the bone tissue, is a complex orthopedic condition marked by chronic inflammation, diagnostic uncertainty, and recurrent infections. Despite standard treatments—including surgical debridement, antimicrobial therapy, and bone reconstruction—many patients continue to experience recurrence and treatment failure. Growing molecular [...] Read more.

Osteomyelitis (OM), an inflammatory condition of the bone tissue, is a complex orthopedic condition marked by chronic inflammation, diagnostic uncertainty, and recurrent infections. Despite standard treatments—including surgical debridement, antimicrobial therapy, and bone reconstruction—many patients continue to experience recurrence and treatment failure. Growing molecular evidence indicates that host genetic factors play a crucial role in shaping immune responses and influencing disease progression in OM. This narrative review synthesizes current knowledge from candidate gene single-nucleotide polymorphism (SNP) association studies to illustrate how specific genetic variations contribute to OM pathogenesis, diagnostic refinement, and treatment outcomes. We examined key immunogenetic variants within genes involved in inflammatory signaling, pathogen recognition, and neutrophil regulation. Our synthesis identifies a landscape of pro-inflammatory SNPs, such as IL-1β rs16944 and NLRP3 rs10754558, that are associated with increased susceptibility to chronic or post-traumatic OM, as well as SNPs that are associated with protective effects that may favor infection resolution, such as within the NOS2 and VDR genes. These SNP-driven differences in inflammasome activity, cytokine pathways, and oxidative stress responses highlight emerging opportunities for individualized therapeutic strategies. This review consolidates these variants, providing a genetic framework to analyze host susceptibility and differentiating high risk from protective genetic profiles. Integrating genomic insights into OM management represents a promising shift toward personalized medicine, enhancing diagnostic precision, informing targeted interventions, and improving prognostic assessment. Continued large-scale validation of candidate SNPs and translational genomic models will be essential to support their future clinical application. Full article

(This article belongs to the Section Antibiotic Therapy in Infectious Diseases)

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31 pages, 5095 KB

Open AccessSystematic Review

Prevalence of Carbapenem-Resistant Gram-Negative Bacilli from Intensive Care Units from Latin America and the Caribbean: A Systematic Review and Meta-Analysis

by Jesús D. Rojas, Mercy Carolina Merejildo Vera, Juan Carlos Benites Azabache, Valeria De La Cruz Surco, Juan Raúl Lucas López and Rafael Pichardo-Rodriguez

Antibiotics 2026, 15(2), 209; https://doi.org/10.3390/antibiotics15020209 - 14 Feb 2026

Cited by 2 | Viewed by 1836

Abstract

Background: Carbapenem-resistant Gram-negative bacilli (CR-GNB) represent a critical threat to patients in intensive care units (ICUs), where limited therapeutic options contribute to elevated mortality. In Latin America and the Caribbean (LAC), the epidemiological burden of CR-GNB remains insufficiently characterized due to fragmented surveillance [...] Read more.

Background: Carbapenem-resistant Gram-negative bacilli (CR-GNB) represent a critical threat to patients in intensive care units (ICUs), where limited therapeutic options contribute to elevated mortality. In Latin America and the Caribbean (LAC), the epidemiological burden of CR-GNB remains insufficiently characterized due to fragmented surveillance systems. This systematic review aimed to synthesize pooled prevalence estimates of CR-GNB among Gram-negative isolates recovered from ICUs across LAC countries. Methods: This systematic review was prospectively registered with PROSPERO (CRD420251177826), followed PRISMA 2020 guidelines and the JBI Manual for prevalence reviews. We searched PubMed, Scopus, LILACS, and SciELO from January 2015 to October 2025 without language restrictions. Observational studies reporting phenotypic carbapenem resistance data from ICUs in LAC countries were eligible. Two reviewers independently screened studies and extracted data. A two-level multilevel generalized linear mixed model (GLMM) with logit transformation was employed using a random-effects approach. Risk of bias was assessed using the JBI critical appraisal checklist. Results: Of 25 eligible studies spanning eight countries, 12 contributed 49 observations to quantitative synthesis. Overall pooled prevalence of CR-GNB was 28.88% (95% CI: 17.32–44.05%), with considerable heterogeneity (I² = 95.24%). Species-specific prevalence was highest for Acinetobacter baumannii (72.58%), followed by Klebsiella pneumoniae (37.48%) and Pseudomonas aeruginosa (29.93%). Regional stratification revealed higher prevalence in South America (31.81%) compared to North America (22.65%) and the Caribbean (11.63%). Conclusions: Nearly one-third of Gram-negative isolates from LAC ICUs exhibit carbapenem resistance, with A. baumannii predominating. Substantial inter-study heterogeneity underscores the need for standardized regional surveillance networks and coordinated antimicrobial stewardship initiatives. Full article

(This article belongs to the Special Issue Antimicrobial Resistance and Epidemiological Study of Clinically Relevant Pathogens)

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23 pages, 11094 KB

Open AccessArticle

Structure–Activity Relationship and Biosafety of Linear Pentapeptide Analogs Derived from Battacin for Antimicrobial Development

by Haixin Sun, Yujie Zhang, Guoqing Gi and Chen Yao

Antibiotics 2026, 15(2), 208; https://doi.org/10.3390/antibiotics15020208 - 13 Feb 2026

Viewed by 774

Abstract

Background: Natural antimicrobial peptides (AMPs) present a promising solution to address the global threat of drug-resistant infections; however, their clinical translation is challenged by limitations in stability, cytotoxicity, and production costs. Methods: In the present study, a linear Battacin-derived peptide (DDLFD) was modified [...] Read more.

Background: Natural antimicrobial peptides (AMPs) present a promising solution to address the global threat of drug-resistant infections; however, their clinical translation is challenged by limitations in stability, cytotoxicity, and production costs. Methods: In the present study, a linear Battacin-derived peptide (DDLFD) was modified at the N-terminus with lipid chains, cinnamic acid, or lipoic acid. The lipoic acid-modified variant was further crosslinked by UV irradiation to form stable nanoparticles. The antibacterial performance against planktonic and biofilm bacteria was systematically evaluated in vitro. Results: The results demonstrated that lauric acid-modified pentapeptide (C₁₂-5) and crosslinked lipoic acid-modified pentapeptide (cLA-5) exhibited potent and rapid-acting effects against various pathogens, including methicillin-resistant Staphylococcus aureus (MRSA). Moreover, they showed enhanced efficacy in eradicating bacterial biofilms. Biosafety assessments based on hemolysis and cytotoxicity assays indicated favorable biocompatibility profiles of cLA-5. Mechanistic investigations confirmed that the modified pentapeptides retained a membrane-targeting mode of action characteristic of natural AMPs, involving membrane depolarization and increased permeability. This physical mechanism effectively prevented the development of resistance in sequential passaging assays and showed strong synergistic effects with ciprofloxacin against ciprofloxacin-resistant strains, effectively restoring their antibiotic susceptibility. Conclusions: Together, these findings underscore the strategic potential of rational structural modification, especially the crosslinked nanostructure, in advancing engineered AMPs toward clinical application. Full article

(This article belongs to the Special Issue Multitalented Synthetic Antimicrobial Compounds: From Design to Effect)

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19 pages, 805 KB

Open AccessArticle

Trends of Microorganisms and Antibiotic Resistance Isolated from Patients with Bacterial Keratitis from a Tertiary Hospital in Southeastern Korea: A 26-Year Retrospective Medical Record Review

by Chan-Ho Cho, Jong Ho Lee and Sang-Bumm Lee

Antibiotics 2026, 15(2), 207; https://doi.org/10.3390/antibiotics15020207 - 13 Feb 2026

Cited by 2 | Viewed by 1116

Abstract

Background. The aim of this study is to analyze changing trends in isolated organisms and antibiotic resistance of bacterial keratitis (BK) over 26 years. Methods. A retrospective medical record review included 542 strains isolated from 462 BK patients between 1998 and 2023. We [...] Read more.

Background. The aim of this study is to analyze changing trends in isolated organisms and antibiotic resistance of bacterial keratitis (BK) over 26 years. Methods. A retrospective medical record review included 542 strains isolated from 462 BK patients between 1998 and 2023. We analyzed routinely generated in vitro antibiotic susceptibility testing results recorded in the laboratory information system and did not perform additional susceptibility testing for research purposes. The entire period was divided into two (first half: 1998–2010, 297 isolates from 255 patients; second half: 2011–2023, 245 isolates from 207 patients) and compared. Results. During the entire period, Staphylococcus spp. (32.3%) and Pseudomonas spp. (18.1%) were common isolates, and a significant increase in Acinetobacter spp. (1.3% vs. 10.6%, p < 0.001) was observed. Among Gram-positive bacteria, methicillin resistance rates remained stable between the two periods (52.6% vs. 46.7%, p = 0.525), and an increase in vancomycin-resistant Enterococci (VRE, 0% vs. 26.1%, p = 0.074) was found. Among Gram-negative bacteria (GNB), ciprofloxacin (7.5% vs. 14.4%, p = 0.108) and imipenem (2.9% vs. 6.5%, p = 0.255) resistance increased slightly, resistance to ceftazidime (8.3% vs. 8.8%, p > 0.999) was maintained, and resistance to aminoglycosides (17.8% vs. 7.2%, p = 0.010) decreased. Conclusions. Our study suggests that conventional topical fortified antibiotic eye drops (tobramycin, ceftazidime) can still be considered as an empirical treatment option for BK. However, our findings revealed a long-term trend of increasing Acinetobacter spp. and VRE, as well as a slight trend of increasing resistance to ciprofloxacin and imipenem in GNB, which may present future challenges in BK treatment. Full article

(This article belongs to the Collection Antibiotics in Ophthalmology Practice)

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20 pages, 5325 KB

Open AccessReview

Neonatal Listeriosis with Central Nervous System Involvement: A Case Series and Review of the Literature

by Chiara Maddaloni, Ludovica Martini, Domenico Umberto De Rose, Daniela Longo, Alessia Guarnera, Cinzia Auriti, Francesca Campi, Maria Paola Ronchetti and Andrea Dotta

Antibiotics 2026, 15(2), 206; https://doi.org/10.3390/antibiotics15020206 - 13 Feb 2026

Cited by 1 | Viewed by 1873

Abstract

Background: Listeria monocytogenes infection during pregnancy remains an underrecognized cause of severe neonatal disease, frequently leading to central nervous system (CNS) involvement with high mortality and long-term neurological sequelae. Case presentation: We report a case series of four neonates with confirmed neonatal listeriosis [...] Read more.

Background: Listeria monocytogenes infection during pregnancy remains an underrecognized cause of severe neonatal disease, frequently leading to central nervous system (CNS) involvement with high mortality and long-term neurological sequelae. Case presentation: We report a case series of four neonates with confirmed neonatal listeriosis and neurological complications, managed in a tertiary neonatal intensive care unit (NICU). Clinical features, microbiological findings, neuroimaging, treatments, and outcomes were analyzed. Our cases presented with early-onset disease and severe clinical courses, including sepsis, meningitis, ventriculitis, hydrocephalus, and seizures. Neuroimaging revealed extensive CNS injury, ranging from intraventricular hemorrhage to multiloculated hydrocephalus. Outcomes varied from near-normal neurodevelopment to profound neurological impairment, despite appropriate antimicrobial therapy. A narrative review of previous cases of neonatal listeriosis was also performed to contextualize our findings. Conclusions: Neonatal listeriosis remains associated with severe neurological morbidity. Early recognition, advanced neuroimaging, multidisciplinary management, and preventive maternal strategies are essential to improve outcomes. Full article

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13 pages, 253 KB

Open AccessArticle

Study on the Importance of Hygienic Hand Disinfection of Dental Practitioners and Students as an Infection Control Measure in Dental Practice

by Veselina Kondeva, Velina Stoeva, Yordan Kalchev and Rumyana Stoyanova

Antibiotics 2026, 15(2), 205; https://doi.org/10.3390/antibiotics15020205 - 13 Feb 2026

Cited by 2 | Viewed by 1032

Abstract

The hands of dental students and practitioners are a key epidemiological factor in the transmission of infections associated with dental care. Strict adherence to the established hand hygiene protocols, combined with regular training and monitoring the quality of the performed hygienic hand disinfection, [...] Read more.

The hands of dental students and practitioners are a key epidemiological factor in the transmission of infections associated with dental care. Strict adherence to the established hand hygiene protocols, combined with regular training and monitoring the quality of the performed hygienic hand disinfection, is crucial for ensuring safe dental practice. Objectives: The aim of the study is to assess the quality of hand antisepsis performed with alcohol-containing preparation among dental students and practicing dentists. Methods: A prospective epidemiological and microbiological study was conducted on 225 people—149 students from the 4th, 5th and 6th year of training at the Faculty of Dental Medicine, Plovdiv, and 76 dentists. The skin antiseptic was applied according to the “six steps” method with alcohol-based antiseptics. The samples were taken with a dry sterile swab. Results: The comparison between students and practicing dental medical doctors shows that the latter have a higher relative share of samples with microbiological growth 12 (15.8%), including coagulase-negative staphylococci (CoNS) above 10⁵ CoNS, compared to students 6 (4.0%), (p = 0.004). Gram-negative microbiological isolates indicate a statistically significant gender dependence (p = 0.016)—15 in men (15.8%), compared to 7 in women (5.4%). Growth of fungi (yeasts and mols) is statistically significant depending on gender (p = 0.015) and is observed only in men. Conclusions: The presence of significant microbial counts of CoNS is an indicator of insufficiently effective hygienic hand disinfection. The recovery of Gram-negative enteric bacteria is unacceptable and suggests serious shortcomings in the hygienic disinfection of some of the samples studied. Students demonstrated superior hand antisepsis performance compared to practicing dentists. Full article

(This article belongs to the Special Issue Evidence-Based Antibiotic Protocols and Digital Innovation in Oral Infection Management: Optimizing Use Through Teledentistry and Modern Diagnostics)

17 pages, 3310 KB

Open AccessArticle

Examining the Effects: Lack of Impact by Endolysin and Phage Treatment on Rotifer and Larvae Microbiota

by Jaime Romero, Carolina Ramírez, Alda Pardo, Marco Medina-Morillo, Luz Hurtado, Rodrigo Rojas and Claudio D. Miranda

Antibiotics 2026, 15(2), 204; https://doi.org/10.3390/antibiotics15020204 - 13 Feb 2026

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Abstract

Background: Bacteriophages and phage-derived lytic enzymes are increasingly considered to be targeted antimicrobial tools in aquaculture; however, their compatibility with non-target microbial communities under hatchery-relevant conditions remains insufficiently characterized. Objectives This study evaluates the impact of a lytic phage (CH20) and a phage-derived [...] Read more.

Background: Bacteriophages and phage-derived lytic enzymes are increasingly considered to be targeted antimicrobial tools in aquaculture; however, their compatibility with non-target microbial communities under hatchery-relevant conditions remains insufficiently characterized. Objectives This study evaluates the impact of a lytic phage (CH20) and a phage-derived lysin (LysVp1), applied under previously validated conditions for rapid Vibrio control, on the microbiota associated with seawater, rotifers, and zebrafish larvae challenged with Vibrio alginolyticus GV09. Methods: Treatments were independently applied to each biological matrix using short exposure times representative of hatchery practices, intentionally capturing the critical window during which microbial transfer from live feed to larvae occurs. Microbial communities were analyzed using 16S rRNA gene sequencing, with DNA- and RNA-derived datasets evaluated separately. Results: Alpha diversity indices were compared using appropriate statistical tests, while beta diversity was assessed using Aitchison distance, PERMANOVA, and dispersion analyses, and differential abundance was evaluated using ANCOM-BC2. Alpha diversity metrics showed no significant differences among treatments across all matrices, indicating the preservation of microbial richness and diversity. Beta diversity patterns differed according to the nucleic acid source, with RNA-based analyses revealing treatment-associated shifts in rotifer and water microbiota that were not consistently detected at the DNA level. In zebrafish larvae, neither phage nor lysin treatment significantly altered overall community structure, although dispersion effects reflected limitations related to sample size. Conclusions: Overall, these results indicate that phage CH20 and lysin LysVp1 exert minimal impact on alpha diversity and limited, context-dependent effects on microbial community structure, supporting their microbiota-safe potential for aquaculture applications. Full article

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Antibiotics, Volume 15, Issue 2 (February 2026) – 122 articles

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