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Bacteriophages (phages) may constitute a natural, safe, and effective strategy to prevent and control multidrug-resistant organisms (MDROs), and ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) pathogens in particular. Few clinical studies have assessed the safety and efficacy of phages in patients infected with MDROs. This systematic review summarizes and critically evaluates published studies of phages in clinical practice and presents the appropriate phage selection criteria, as well as recommendations for clinicians and scientists for a successful therapy. Articles were identified through a search of the PubMed, Ovid, EMBASE, and Cochrane Library databases. Among 1102 articles and abstracts, 30 studies were selected and evaluated using selective inclusion criteria, phage criteria, and study characteristics. Most studies showed efficacy (87%) and safety (67%) of the tested phages, but few studies examined phage resistance (35%). Clinical studies and regulatory changes are needed to determine the safety and efficacy of phages and to advance their use in patients with MDRO infections.

Antimicrobial resistance is a major threat for public health. Plasmids play a critical role in the spread of antimicrobial resistance via horizontal gene transfer between bacterial species. However, it remains unclear how plasmids originally recruit and assemble various antibiotic resistance genes (ARGs). Here, we track ARG recruitment and assembly in clinically relevant plasmids by combining a systematic analysis of 2420 complete plasmid genomes and experimental validation. Results showed that ARG transfer across plasmids is prevalent, and 87% ARGs were observed to potentially transfer among various plasmids among 8229 plasmid-borne ARGs. Interestingly, recruitment and assembly of ARGs occur mostly among compatible plasmids within the same bacterial cell, with over 88% of ARG transfers occurring between compatible plasmids. Integron and insertion sequences drive the ongoing ARG acquisition by plasmids, especially in which IS26 facilitates 63.1% of ARG transfer events among plasmids. In vitro experiment validated the important role of IS26 involved in transferring gentamicin resistance gene aacC1 between compatible plasmids. Network analysis showed four beta-lactam genes (blaTEM-1, blaNDM-4, blaKPC-2, and blaSHV-1) shuffling among 1029 plasmids and 45 clinical pathogens, suggesting that clinically alarming ARGs transferred accelerate the propagation of antibiotic resistance in clinical pathogens. ARGs in plasmids are also able to transmit across clinical and environmental boundaries, in terms of the high-sequence similarities of plasmid-borne ARGs between clinical and environmental plasmids. This study demonstrated that inter-plasmid ARG transfer is a universal mechanism for plasmid to recruit various ARGs, thus advancing our understanding of the emergence of multidrug-resistant plasmids.

Antimicrobial resistance is now considered a major global challenge; compromising medical advancements and our ability to treat infectious disease. Increased antimicrobial resistance has resulted in increased morbidity and mortality due to infectious diseases worldwide. The lack of discovery of novel compounds from natural products or new classes of antimicrobials, encouraged us to recycle discontinued antimicrobials that were previously removed from routine use due to their toxicity, e.g., colistin. Since the discovery of new classes of compounds is extremely expensive and has very little success, one strategy to overcome this issue could be the application of synthetic compounds that possess antimicrobial activities. Polymers with innate antimicrobial properties or that have the ability to be conjugated with other antimicrobial compounds create the possibility for replacement of antimicrobials either for the direct application as medicine or implanted on medical devices to control infection. Here, we provide the latest update on research related to antimicrobial polymers in the context of ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) pathogens. We summarise polymer subgroups: compounds containing natural peptides, halogens, phosphor and sulfo derivatives and phenol and benzoic derivatives, organometalic polymers, metal nanoparticles incorporated into polymeric carriers, dendrimers and polymer-based guanidine. We intend to enhance understanding in the field and promote further work on the development of polymer based antimicrobial compounds.

Multi‐Locus Sequence Typing (MLST) is a key method for allocation of Sequence Types (STs) for bacterial isolates. Traditionally, this is performed by the Sanger sequencing method, which can be highly time‐consuming and laborious. In this study, we present NanoMLST, a high‐throughput MLST workflow using multiplex PCR, Oxford Nanopore Technologies Next‐Generation Sequencing, and the Krocus program for typing ESKAPE + E pathogens (Enterococcus faecium [E. faecium], Staphylococcus aureus, Klebsiella pneumoniae [K. pneumoniae], Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp., and Escherichia coli). Bacterial isolates were obtained from the Hospital Universitario La Paz's Microbiology Department and the Centro Nacional de Microbiología. Primers that can be multiplexed in a single PCR reaction were designed for the seven housekeeping genes for each species. DNA was extracted from single colonies by heating at 95°C for 10 min, mechanical lysis at 4.20 m/s for 2 min, and then by the MagCore extraction system. Multiplex PCRs were then performed with the respective primer mixes for each species, and libraries were prepared for sequencing by ONT Flongle cells. The Krocus program was then used to determine the STs from the raw FastQ reads. STs for 221 isolates were obtained through this workflow with an average time of 12 h per 24 isolates. In line with local data, the K. pneumoniae and E. faecium isolates were relatively oligoclonal, while the rest were polyclonal. STs from representative isolates showed 100% concordance between Sanger sequencing and the proposed workflow. NanoMLST offers a fast, cheaper, and less labor‐intensive alternative for large‐scale MLST applications targeting clinically important pathogens. NanoMLST is a workflow suggested as a viable alternative to high‐throughput ST determination for ESKAPE + E pathogens using ONT next‐generation sequencing.

In the modern era, the expanding demand for implants has transformed the healthcare system by restoring and enhancing the function of various biological structures, thereby increasing the patients’ quality of life. These include urinary catheters, dental, orthopedic, cardiovascular implants, and sutures designed to perform various functions. However, these devices are more prone to microbial attack, contributing to biofilm formation mainly caused by multidrug-resistant ESKAPE pathogens, thereby increasing the risk of implant-associated infections and implant failure. This review summarizes the diverse array of implants available on the market and their associated infections caused by biofilm-producing pathogens, with a particular emphasis on the ESKAPE pathogen. Specific keywords were used to conduct a literature review using Google Scholar, Web of Science, PubMed, and Scopus databases. The data were then screened and integrated to explore the underlying principles of biofilm formation, its consequences, diagnostic approaches, and therapeutic studies. Currently, various methods are employed to diagnose these infections, including culture-based methods (tissue swab, culture, sonication) and non-culture methods (Dithiothreitol, XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide), Resazurin, BioTimer assays, and PCR). However, these studies indicate an increased difficulty in detecting infections caused by ESKAPE pathogens due to biofilm formation, highlighting the need for developing novel strategies. The recent advancements in the development of antimicrobial coatings, implant surface modifications, phage therapy, nanoparticles, antimicrobial peptides, and quorum-sensing inhibitors have shown promise in controlling these infections. Thus, these findings underscore the importance of research on innovative approaches and the development of infection-resistant implants, thereby reducing the clinical burden and improving patient outcomes.

The article explores drug-resistant bacteria within the ESKAPE group, commonly associated with nosocomial infections, focusing on the resistance mechanisms of Acinetobacter baumannii and Klebsiella pneumoniae. The study delves into various β-lactamase enzymes and resistance mechanisms exhibited by ESKAPE bacteria, shedding light on the challenges posed by carbapenem-resistant infections. Notably, the article underscores the ongoing need for research to develop more effective treatments and address the persistent challenges associated with drug resistance in the context of nosocomial infections. The examination of this subset of bacteria aims to contribute to a comprehensive understanding of their resistance mechanisms and provides insights into the difficulties encountered in treating infections with carbapenem-resistant pathogens. The article serves as a valuable resource for clinicians, researchers and policymakers, offering a detailed perspective on the current state of drug resistance among nosocomial pathogens and advocating for continuous research to enhance treatment efficacy in the face of evolving challenges.

In recent years, the Infectious Diseases Society of America has highlighted a faction of antibiotic-resistant bacteria (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) - acronymically dubbed 'the ESKAPE pathogens' - capable of 'escaping' the biocidal action of antibiotics and mutually representing new paradigms in pathogenesis, transmission and resistance. This review aims to consolidate clinically relevant background information on the ESKAPE pathogens and provide a contemporary summary of bacterial resistance, alongside pertinent microbiological considerations necessary to face the mounting threat of antimicrobial resistance.

Background: Health care-associated infections (HAIs) are a significant public health problem worldwide, favoring multidrug-resistant (MDR) microorganisms. The SARS-CoV-2 infection was negatively associated with the increase in antimicrobial resistance, and the ESKAPE group had the most significant impact on HAIs. The study evaluated the bactericidal effect of a high concentration of O3 gas on some reference and ESKAPE bacteria. Material and Methods: Four standard strains and four clinical or environmental MDR strains were exposed to elevated ozone doses at different concentrations and times. Bacterial inactivation (growth and cultivability) was investigated using colony counts and resazurin as metabolic indicators. Scanning electron microscopy (SEM) was performed. Results: The culture exposure to a high level of O3 inhibited the growth of all bacterial strains tested with a statistically significant reduction in colony count compared to the control group. The cell viability of S. aureus (MRSA) (99.6%) and P. aeruginosa (XDR) (29.2%) was reduced considerably, and SEM showed damage to bacteria after O3 treatment Conclusion: The impact of HAIs can be easily dampened by the widespread use of ozone in ICUs. This product usually degrades into molecular oxygen and has a low toxicity compared to other sanitization products. However, high doses of ozone were able to interfere with the growth of all strains studied, evidencing that ozone-based decontamination approaches may represent the future of hospital cleaning methods.

An increasing number of multidrug-resistant pathogens is a serious problem of modern medicine and new antibiotics are highly demanded. In this study, different n-alkyl acids (C2-C14) and aromatic acids (benzoic and trans-cinnamic) were conjugated to the N-terminus of KR12 amide. The effect of this modification on antimicrobial activity (ESKAPE bacteria and biofilm of Staphylococcus aureus) and cytotoxicity (human red blood cells and HaCaT cell line) was examined. The effect of lipophilic modifications on helicity was studied by CD spectroscopy, whereas peptide self-assembly was studied by surface tension measurements and NMR spectroscopy. As shown, conjugation of the KR12-NH2 peptide with C4-C14 fatty acid chains enhanced the antimicrobial activity with an optimum demonstrated by C8-KR12-NH2 (MIC 1–4 μg/mL against ESKAPE strains; MBEC of S. aureus 4–16 μg/mL). Correlation between antimicrobial activity and self-assembly behavior of C14-KR12-NH2 and C8-KR12-NH2 has shown that the former self-assembled into larger aggregated structures, which reduced its antimicrobial activity. In conclusion, N-terminal modification can enhance antimicrobial activity of KR12-NH2; however, at the same time, the cytotoxicity increases. It seems that the selectivity against pathogens over human cells can be achieved through conjugation of peptide N-terminus with appropriate n-alkyl fatty and aromatic acids.

Airborne microorganisms significantly contribute to hospital-associated infections (HAIs), particularly causing respiratory tract infections (RTIs). The spread of Multi-Drug Resistant (MDR) airborne bacteria further complicates disease prevention, challenging existing infection control strategies. Current air decontamination technologies are found to have limitations, necessitating novel approaches. In this study, we demonstrated the utility of zeta potential, a natural physicochemical electro-kinetic property of bacteria as a key target, which can be exploited to trap and eliminate MDR airborne bacteria. Multiple respiratory pathogens were included in the study, harbouring various resistance phenotypes. Zeta potentials of these clinical isolates were measured and compared against corresponding ATCC strains. Clinical isolates were aerosolized in a certified BSL-2 setting containing a ZeBox-powered air sterilization device. Viable bacteria were enumerated at various time points, before and after exposure to the air decontamination device. Our analyses revealed that Zeta potential is relatively independent of the origin and antibiotic susceptibility of the tested isolates. Exposure to ZeBox powered device for 5 min resulted in a minimum of 5 log reduction (99.999%) among majority of the isolates, irrespective of their genus and origin. Zeta potential measurements correlated to the kill kinetics of ZeBox technology. The current study underscores the reliability of zeta potential based air decontamination technologies such as ZeBox for potential elimination of diverse, airborne respiratory pathogens in healthcare and domestic settings, offering a promising strategy to combat HAIs in post antibiotic and post pandemic era.