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Antibiotics, Volume 15, Issue 6 (June 2026) – 5 articles

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21 pages, 1042 KB

Open AccessArticle

Multiplex PCR-Based Detection of Eight Carbapenemase Genes and Their Clinical Characteristics in Urinary Tract Infections

by Nishadi Jayathilaka, Upeksha Kulasekara, Dilini Nakkawita, Dharshan De Silva, Samanmalee Gunasekara and Thamarasi Senaratne

Antibiotics 2026, 15(6), 529; https://doi.org/10.3390/antibiotics15060529 (registering DOI) - 22 May 2026

Abstract

Background: The emergence and spread of urinary carbapenem-resistant organisms (CROs) are a major public health concern, particularly in Sri Lanka. Therefore, we aimed to detect and genotypically characterize CROs in urinary tract infections (UTIs) and their clinical outcomes. Methods: Urinary CROs were collected from two hospitals in Sri Lanka from January to December, 2023. Among 7640 urine samples, 100 CROs were identified by disk diffusion method, and 99 were detected by BD Pheonix^TM automated system. The presence of eight carbapenemase genes; bla_KPC, bla_NDM, bla_VIM, bla_IMP, bla_OXA-23, bla_OXA-48, bla_OXA-51, and bla_OXA-58, among 97 CROs was detected by a multiplex PCR kit. Results: Out of 99 urinary CROs, K. pneumoniae (33.3%; n = 33/97) was the most common species. Among the 97 isolates tested by PCR, a single carbapenemase gene was detected in 35.05% (34/97), while two or more genes co-occurred in 39.18% (38/97). The most frequently identified gene was bla_OXA-51 (47.4%), followed by bla_OXA-58 (41.2%). Most patients (95.74%; n = 90/97) showed clinical improvement within seven days of treatment. Among the 93 patients discharged and followed for three months, 74.20% (n = 69/93) experienced at least one mild UTI recurrence. A total of 10 patients died during the study period. Of which, four (40%) during hospitalization and six (60%) during follow-up, though none of the deaths were attributed to UTIs. Conclusions: K. pneumoniae, showed the highest carbapenemase gene diversity. Recurrent UTIs were observed during the follow-up period. Continuous surveillance and implementation of targeted infection control programs are needed to minimize further emergence and spread of carbapenemase genes. Full article

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

23 pages, 3382 KB

Open AccessArticle

Epidemiology of Clostridioides difficile Infection in Argentina and Associated Risk Factors Evaluated Through a Meta-Analysis

by Angela María Barbero, Nicolás Diego Moriconi, Sabina Palma, Josefina Celano, María Gracia Balbi, Lorenzo Sebastián Morro, María Martina Calvo Zarlenga, Jorgelina Suárez, María Guadalupe Martínez, Mónica Graciela Machain, Carlos Gabriel Altamiranda, Gabriel Erbiti, Rodrigo Emanuel Hernández Del Pino and Virginia Pasquinelli

Antibiotics 2026, 15(6), 528; https://doi.org/10.3390/antibiotics15060528 (registering DOI) - 22 May 2026

Abstract

Background: Clostridioides difficile is classified within the first 18 threats for antimicrobial resistance and is the leading cause of hospital-acquired enteric infection. Community-associated cases have notably increased in recent decades, highlighting that accurate and up-to-date statistics characterizing the epidemiology of C. difficile infection (CDI) are critical. Methods: We conducted a retrospective (2019–2023) case-control study evaluating the prevalence of CDI in 249 stool samples from hospitalized patients in the sanitary region III of Buenos Aires, Argentina. The presence of C. difficile was detected by combining EIA, PCR, and toxigenic culture via a diagnostic algorithm. Clinical and demographic data from patients were analyzed to identify CDI-associated risk factors. We also conducted a systematic review and a meta-analysis contrasting our results with 38 studies selected from different countries. Results: One in five patients presented C. difficile as the etiological agent of diarrhea. Eighty percent of the CDI+ cases carried toxigenic strains, with a third of cases associated with community environments. Age ≥ 69 years, previous use of antibiotics, previous hospitalization, and previous episodes of CDI emerged as predisposing factors for CDI in our study cohort. In an exploratory evaluation of clinical data, CDI+ patients showed higher leukocytes and platelets counts, a decreased basophil count, and increased urea concentration. At the global level, the meta-analysis reinforced advanced age, previous consumption of antibiotics, previous consumption of proton pump inhibitors, previous hospitalization, and previous CDI as risk factors for CDI. Conclusions: These results emphasize the importance of continued epidemiological surveillance of CDI. Our findings confirm previously described risk factors, both in our cohort and at the global level. Exploratory alterations in laboratory parameters were observed, although their clinical relevance and specificity require further investigation. Full article

(This article belongs to the Special Issue Clostridioides difficile Infection, 3rd Edition)

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19 pages, 1717 KB

Open AccessArticle

Antarctic Fish Antimicrobial Peptides Active Against Bacterial and Viral Pathogens of Aquacultural Importance

by Federica Massaro, Luana Cortinovis, Romy Lucon Xiccato, Eleonora Fiocchi, Amedeo Manfrin, Anna Rita Taddei, Paolo Roberto Saraceni, Fernando Porcelli, Anna Toffan and Francesco Buonocore

Antibiotics 2026, 15(6), 527; https://doi.org/10.3390/antibiotics15060527 - 22 May 2026

Abstract

Background/Objectives: The aquaculture industry represents a fundamental food sector. One of the main limiting factors for this sector is related to bacterial diseases, for which antibiotics have been widely used worldwide for decades. In recent years, a more conscious approach to the use of antimicrobials within the framework of the One Health approach has increased the need for alternatives capable of helping with disease management while avoiding the onset of antimicrobial resistance phenomena. Antimicrobial peptides, which have a broad spectrum of action against pathogens, are a promising solution. Methods: In this work, we investigated the capability of three peptides (Trematocine, Chionodracine, and Cnd-m3) isolated from Antarctic fish to target bacterial and viral pathogens affecting aquaculture. Successively, we investigated their cytotoxicity versus a continuous embryonic cell line (DLEC) derived from European sea bass and their haemolytic activity against fish erythrocytes. Moreover, we evaluated their immunomodulatory effect. Results: Regarding antibacterial properties, Cnd-m3 was identified as the best peptide, demonstrating good bactericidal and bacteriostatic activity against various bacterial strains, including Lactococcus garvieae. Concerning this bacterium, ANS permeability assays showed that the Cnd-m3 peptide has a great ability to interact with its outer membrane, while TEM analysis revealed that the peptide, after destabilization of the cell membrane, interacts with nucleic structures. Considering the antiviral activity, Trematocine was effective against two tested pathogenic enveloped viruses. Moreover, the toxicity of Trematocine and Cnd-m3 was evaluated by investigating their cytotoxicity against a cell line derived from Dicentrarchus labrax and haemolysis against sea bass erythrocytes. Both revealed good selectivity towards pathogens at the lowest concentration. Finally, Cnd-m3 manifested light in vitro immunomodulatory properties. Conclusions: Overall, these data provide a solid basis for future studies assessing the potential applications of two of the tested peptides in aquaculture. Full article

(This article belongs to the Section Antimicrobial Peptides)

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25 pages, 698 KB

Open AccessReview

Bacterial Persister Cells as Evolutionary Catalysts of Antibiotic Resistance: Mechanisms, Clinical Implications, and Therapeutic Strategies

by Tae-Jong Kim

Antibiotics 2026, 15(6), 526; https://doi.org/10.3390/antibiotics15060526 - 22 May 2026

Abstract

Antibiotic resistance is a growing global health threat. However, its evolution cannot be fully understood without considering antibiotic tolerance and persistence. Persister cells are phenotypic variants that survive lethal antibiotic exposure without heritable resistance, primarily through growth arrest, metabolic slowdown, and stress-adaptive states. Although persistence has been viewed as a transient survival phenomenon, increasing evidence suggests that it may also have a genetic basis by preserving populations during antibiotic-induced bottlenecks and enabling regrowth, mutation, and selection under certain conditions. This review examines the molecular mechanisms underlying persister formation, including toxin–antitoxin systems, stringent-response signaling, ATP depletion, translational arrest, and stress-response networks. We discuss how persistence contributes to antibiotic tolerance in biofilms, host environments, and recurrent infections, and how repeated antibiotic exposure may promote stepwise evolution from phenotypic survival to stable resistance in specific contexts. Evidence from experimental evolution, clinical observations, and system-level analyses supports a potential but context-dependent link between persistence and resistance. We also highlight therapeutic strategies targeting persister cells, including antipersister compounds, metabolic activation, combination therapies, bacteriophages, and alternative approaches. Finally, we outline future research directions, emphasizing single-cell technologies, systems biology, longitudinal clinical studies, and evolution-informed treatment design. A comprehensive understanding of persistence and its evolutionary implications is essential for improving treatment efficacy and limiting the emergence of long-term antibiotic resistance. Full article

(This article belongs to the Special Issue Feature Reviews in Mechanism and Evolution of Antibiotic Resistance 2026)

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13 pages, 1405 KB

Open AccessArticle

Sustainability and Impact of an Antimicrobial Stewardship Program on Broad-Spectrum Antibiotic Consumption in South Korea: A 14-Month Extended Follow-Up Study

by Tae-Hoon No and Kyeong Min Jo

Antibiotics 2026, 15(6), 525; https://doi.org/10.3390/antibiotics15060525 - 22 May 2026

Abstract

Background: Antimicrobial stewardship programs (ASPs) are critical for promoting rational antibiotic use. While early implementation outcomes have been reported, extended follow-up sustainability and the impact on high-priority broad-spectrum antibiotics in South Korean secondary/tertiary hospitals require further validation. This study aimed to evaluate the extended outcomes and sustainability of an ASP over a 14-month period. Methods: This retrospective, single-center study analyzed ASP activities from January 2025 to February 2026 at a tertiary hospital in South Korea. Interventions included prospective audit and feedback (PAF) for restricted antibiotics and recommendations for prolonged prescriptions (≥14 days). Primary outcomes were the monthly rejection rate of restricted antibiotics and the acceptance rate of ASP interventions. Secondary outcomes included the days of therapy (DOT) per 1000 patient–days for meropenem and piperacillin/tazobactam (Pip/Taz). Results: During the 14-month period, the ASP intervention acceptance rate increased significantly from a mean of 72.0% in the implementation phase (January–April 2025) to 81.2% in the stabilization phase (May 2025–February 2026) (p = 0.035). The DOT for Pip/Taz decreased significantly from 169.4 to 151.8 per 1000 patient–days (p = 0.002), with a significant negative correlation identified between the intervention acceptance rate and Pip/Taz consumption (r = −0.625, p = 0.017). Although overall meropenem DOT showed seasonal fluctuations without reaching statistical significance across phases, a year-over-year comparison revealed a 7.5% reduction in meropenem DOT (January–February 2025: 54.8 vs. January–February 2026: 50.7 per 1000 patient–days). The rejection rate for restricted antibiotics declined from 3.8% to 2.6%, suggesting that clinicians increasingly self-regulated inappropriate prescribing attempts. Conclusions: The ASP demonstrated extended follow-up sustainability with a significant reduction in the consumption of key broad-spectrum antibiotics. A progressive increase in clinician acceptance of ASP interventions from 72.0% to 81.2%, combined with a concurrent decline in the restricted antibiotic rejection rate, reflected a measurable shift in institutional prescribing culture and confirmed the successful transition to a stabilized program. These findings support the necessity of sustained multidisciplinary ASPs, even in resource-limited settings, to combat antimicrobial resistance effectively. Full article

(This article belongs to the Special Issue Antibacterial Utilisation from Different Perspectives: An Effective Tool to Develop and Evaluate Antimicrobial Stewardship Interventions)

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