Multidrug-resistant Bacteria Research Articles

First case report of a vertebral osteomyelitis caused by carbapenem-resistant Enterobacter cloacae treated with imipenem/cilastatin/relebactam prolonged infusion then meropenem/vaborbactam in continuous infusion.

Bone and joint infections (BJIs) caused by multidrug-resistant bacteria are becoming more frequent. However, data on the use of novel β-lactam/β-lactamase inhibitors, such as imipenem/cilastatin/relebactam (I-R) and meropenem/vaborbactam (MVB), to treat BJIs is lacking. Furthermore, prolonged infusions of these β-lactams should theoretically optimize pharmacokinetic/pharmacodynamics target in these indications, but there are currently no reports on this type of infusions, especially in the setting of BJI. We report a case of a vertebral osteomyelitis caused by carbapenem-resistant Enterobacter cloacae successfully treated with extended-infusion of I-R (1.25g q6h over 2h), then with continuous infusion of MVB (2g q4h as over 4h). Therapeutic drug monitoring confirmed that extended-infusion of I-R and continuous infusion of MVB achieved serum concentrations up to 12mg/L of imipenem and 19mg/L of meropenem, respectively. The favourable outcome of this patient treated for a vertebral osteomyelitis caused by carbapenem-resistant E. cloacae suggest that extended- and continuous infusions of I-R and MVB, are promising regimens for treatment of BJIs caused by carbapenem-resistant Enterobacterales.

Highly oxygenated dihydrostilbenoids and flavones from the aerial part of Glycyrrhiza uralensis and their antimicrobial activities

Abstract Background Multidrug-resistant bacteria (MDRB) represent a significant global challenge due to their high mortality rates, substantial economic burden, and rapid spread. Traditional triple or quadruple therapies have demonstrated limited efficacy as a result of increasing drug resistance. Thus, it is urgent to develop novel anti-MDRB drugs with high efficiency and low toxicity. Objectives To isolate and identify the dihydrostilbenoids and flavones from the aerial part of Glycyrrhiza uralensis (Fabaceae) and their antimicrobial activities were screened. Materials and methods The aerial part of G. uralensis was extracted with 75% aqueous EtOH. The crude extract was repeatedly isolated by macroporous resin, silica gel, Sephadex LH-20, C18-MPLC, and MCI-MPLC, which were then purified by semipreparative RP-HPLC to obtain monomer compounds. The structures of the isolates were assigned by a combination of optical rotations, UV spectra, nuclear magnetic resonance, and high-resolution electrospray ionization mass spectrometry, and the absolute configurations of compounds 2, 3, and 7 were further confirmed by electronic circular dichroism calculations. Subsequently, we investigated their antimicrobial activities by the broth microdilution method. Results Seventeen previously undescribed phenolic compounds (1–17) and 26 known analogs (18–43), including dihydrostilbenoids, flavones, and dihydroflavones, were identified from the aerial part of G. uralensis. In vitro, antimicrobial bioassays demonstrated that compound 31 displayed the strongest inhibitory effect against 4 drug-resistant Helicobacter pylori strains (MIC = 2–4 μg/mL), comparable to metronidazole (MIC = 1–32 μg/mL). Additionally, compounds 10, 13, and 15 demonstrated bactericidal activity against Staphylococcus aureus (MIC = 4 μg/mL), while compounds 15 and 22 exhibited inactivation effects against Mycobacterium smegmatis, Enterococcus faecium, and E. faecalis (MIC = 4–8 μg/mL). Conclusions These monomeric compounds with antimicrobial activities were isolated from the aerial parts of G. uralensis, providing valuable insights into the potential anti-MDRB properties of its nonmedicinal parts.

Chemical Space for Peptide-based Antimicrobials.

Multidrug-resistant (MDR) bacteria represent a global public health threat, and antimicrobial peptides (AMPs), derived from naturally occurring linear or cyclic peptides, can provide the solution. However, most AMPs are sensitive to proteases and have poor pharmacokinetics. The EU-funded ERC Advanced Grant SPACE4AMPS aims to identify new AMPs by applying the concepts of chemical space and ligand-based virtual screening, which are well known for small molecule drug discovery, to the world of peptides. We create virtual libraries of peptides and related molecules and use these approaches to select a few tens of compounds for synthesis and detailed. evaluation of antibacterial, toxicity and stability effects. Recent results and prospects of this ongoing project are presented in this review.

Klebsiella in Wildlife: Clonal Dynamics and Antibiotic Resistance Profiles, a Systematic Review

Klebsiella spp. are a genus of Gram-negative, opportunistic bacteria frequently found in the flora of the mucosal membranes of healthy animals and humans, and in the environment. Species of this group can cause serious infections (meningitis, sepsis, bacteraemia, urinary tract infections, liver damage) and possible death in immunocompromised organisms (and even in immunocompetent ones in the case of hypervirulent K. pneumoniae) that are exposed to them. K. pneumoniae is part of the ESKAPE organisms, and so it is important to understand this genus in terms of multidrug-resistant bacteria and as a carrier of antibiotic resistance mechanisms. As it is a durable bacterium, it survives well even in hostile environments, making it possible to colonize all kinds of habitats, even the mucosal flora of wildlife. This systematic review explores the prevalence of Klebsiella spp. bacteria in wild animals, and the possibility of transmission to humans according to the One Health perspective. The isolates found in this review proved to be resistant to betalactams (blaTEM, blaOXA-48…), aminoglycosides (strAB, aadA2…), fosfomycin, tetracyclines, sulphonamides, trimethoprim, phenicols (catB4), and polymyxins (mcr4).

Computational prediction of a phage cocktail active against multidrug-resistant bacteria

Background Antibiotic misuse and overuse have contributed to the emergence of multi-drug resistant (MDR) bacteria, posing a serious public health problem across the globe. Phage cocktails, which combine multiple phages, provide an efficient method to combat multidrug-resistant bacterial infections. This study integrated a computational pipeline to design a phage cocktail against the bacterial strains Acinetobacter baumannii AB0057, Klebsiella pneumoniae subsp. pneumoniae HS11286, and Pseudomonas aeruginosa UCBPP-PA14. Methods The whole genome sequences of selected multidrug-resistant bacteria were accessed. Prophage sequences were identified from them which could be expressed to produce viable lytic phages against MDR bacterial strains, thereby reducing the severity of infection. Prophages were annotated for open reading frames (ORFs), putative promoters, virulence factors, transcriptional terminators, ribosomal RNAs, and transfer RNAs. A dot plot was also generated to investigate similar phages and phylogenetic analysis was performed. Results A total of 11 prophages were predicted from the bacterial genomes. About 472 open reading frames were predicted along with 3 transfer RNAs. Additionally, the presence of 754 putative promoters and 281 transcription terminator sequences was also detected. Comparative genomic and phylogenetic analyses provided insight into the diversity, relatedness, and lytic potential of the phages. The final designed phage cocktail consisted of five selected prophages including Acinetobacter baumannii prophages (2759376-2809756) and (3311844-3364667), and Klebsiella pneumoniae prophages (1288317-1338719), (1778306-1808606), and (2280703-2325555). Conclusion The phage cocktail designed in this study might be useful against MDR Acinetobacter baumannii and Klebsiella pneumoniae infections, especially where conventional antibiotics fail. Sequence similarity analysis suggested that the phage cocktail may also be effective against other carbapenemase-producing K. pneumoniae strains.

Selective Digestive Decontamination: A Comprehensive Approach to Reducing Nosocomial Infections and Antimicrobial Resistance in the ICU.

Background/Objective: Multidrug-resistant (MDR) bacteria pose a significant threat to global health, especially in intensive care units (ICUs), where high antibiotic consumption drives antimicrobial resistance. Selective digestive decontamination (SDD) is a strategy designed to prevent nosocomial infections and colonization by MDR pathogens. This study aimed to evaluate the impact of implementing an SDD protocol on antibiotic consumption and colonization by carbapenemase-producing Enterobacterale (CPE) in a specific ICU setting. Methods: This quasi-experimental study was conducted in the ICU of a university hospital from June 2021 to June 2023. Patients were divided into two groups: pre-intervention (before SDD) and post-intervention (after SDD implementation). Data on antibiotic consumption (expressed as defined daily doses (DDDs) per 100 stays), nosocomial infections, colonization rates, and the incidence of MDR bacteria were collected. A statistical analysis was conducted to compare the pre- and post-intervention groups. Results: A total of 3266 patients were included, with 1532 in the pre-intervention group and 1734 in the post-intervention group. The implementation of the SDD protocol resulted in a significant reduction in total antibiotic consumption (p = 0.028), with notable decreases in carbapenem use (p < 0.01) and colonization by CPE (p = 0.0099). The incidence of nosocomial infections also decreased in the post-SDD group, although this reduction was not statistically significant. Conclusions: The implementation of the SDD protocol in this ICU setting significantly reduced antibiotic consumption and colonization by CPE. These findings suggest that SDD may be a valuable tool in managing antimicrobial resistance in critical care settings, without contributing to the development of MDR bacteria.

A recent update on the antibacterial effects ofdistinct bioactive molecules derived from theCannabis plant.

The number of human infections with multidrug-resistant (MDR) bacteria is increasing worldwide and constitutes a serious threat to human health. Given the lack of novel antibiotic compounds worsening this dilemma, alternative antibiotic-independent treatment and prevention strategies of infectious diseases applying natural compounds appear highly appreciable. Given the long-known health-beneficial and disease-alleviating properties of Cannabis, we performed a literature search summarizing current knowledge regarding the antibacterial effects of extracts from different parts of the Cannabis sativa plant and of defined Cannabis-derived molecules and their potential mode of action. Theincluded studies revealed that various extracts and essential oils of C. sativa as well as major cannabinoids exerted potent activities against a broad spectrum of Gram-positive bacteria and against some Gram-negative bacterial species including MDR strains. Particularly the disruption of the bacterial cytoplasmic membrane by some cannabinoids resulted in potent antibacterial effects against Gram-positivebacteria including methicillin-resistant Staphylococcus aureus. Furthermore, defined cannabinoids inhibited the formation of and eradicated existing bacterial biofilms. In conclusion, given their antibacterial properties distinct Cannabis-derived molecules expand the repertoire of antibiotics-independent treatment options in the combat of bacterial infectious diseases which should be further addressed in future studies including clinical trials.

COVID-19 and COVID-19 Vaccine-Related Skin Ulcerations in the Lower Extremities: A Case Report and Literature Review

Several associations have been made between COVID-19 and vasculitis. Recent data also shows the prevalence and association of de novo vasculitis with either COVID-19 infection or COVID-19 post vaccination. In this article, we present the case of new-onset leukocytoclastic vasculitis, secondary to COVID-19 vaccination, that was complicated by severe infected and nonhealing ulcers in the lower extremities. Case A 53-year-old male patient presented to the dermatology clinics with a three-week history of painful necrotic patches coalescent of the lateral malleolus of the right and left ankles. History goes back to when the patient reported developing pruritic papules two weeks after receiving his second shot of the Pfizer BioNTech COVID-19 vaccine (BNT162b2). Punch biopsy was consistent with leukocytoclastic vasculitis. He was prescribed a four-week course of systemic corticosteroids and antibiotics as per cultures. Vascular assessment confirmed normal peripheral arterial and venous system. Two months later, the patient re-presented with fever and worsening of his lower extremity ulcers. He underwent debridement of his wounds. Intra-operative cultures revealed multidrug resistant bacteria. He required an additional debridement session a few days later and a 14-day course of Piperacillin-Tazobactam. The patient was subsequently discharged on corticosteroids and Azathioprine and followed up in the vascular surgery and rheumatology clinics. At four months follow-up, the patient's wounds were almost completely healed. Conclusion This article highlights a case of severe new-onset COVID-19 vaccine-associated leukocytoclastic vasculitis complicated with infected ulcers that required debridement twice in addition to a prolonged course of antibiotics and immunosuppression therapy. To our knowledge, none of the cases reported in the literature were this severe in nature. In this post-pandemic era, it must remain high on the differential list, and healthcare specialists should maintain a high index of suspicion when evaluating sudden new-onset skin lesions that do not have an immediately apparent etiology.

Antimicrobial and antibiofilm activity of human recombinant H1 histones against bacterial infections.

Histones possess significant antimicrobial potential, yet their activity against biofilms remains underexplored. Moreover, concerns regarding adverse effects limit their clinical implementation. We investigated the antibacterial efficacy of human recombinant histone H1 subtypes against Pseudomonas aeruginosa PAO1, both planktonic and in biofilms. After the in vitro tests, toxicity and efficacy were assessed in a P. aeruginosa PAO1 infection model using Galleria mellonella larvae. Histones were also evaluated in combination with ciprofloxacin (Cpx) and gentamicin (Gm). Our results demonstrate antimicrobial activity of all three histones against P. aeruginosa PAO1, with H1.0 and H1.4 showing efficacy at lower concentrations. The bactericidal effect was associated with a mechanism of membrane disruption. In vitro studies using static and dynamic models showed that H1.4 had antibiofilm potential by reducing cell biomass. Neither H1.0 nor H1.4 showed toxicity in G. mellonella larvae, and both increased larvae survival when infected with P. aeruginosa PAO1. Although in vitro synergism was observed between ciprofloxacin and H1.0, no improvement over the antibiotic alone was noted in vivo. Differences in antibacterial and antibiofilm activity were attributed to sequence and structural variations among histone subtypes. Moreover, the efficacy of H1.0 and H1.4 was influenced by the presence and strength of the extracellular matrix. These findings suggest histones hold promise for combating acute and chronic infections caused by pathogens such as P. aeruginosa.IMPORTANCEThe constant increase of multidrug-resistant bacteria is a critical global concern. The inefficacy of current therapies to treat bacterial infections is attributed to multiple mechanisms of resistance, including the capacity to form biofilms. Therefore, the identification of novel and safe therapeutic strategies is imperative. This study confirms the antimicrobial potential of three histone H1 subtypes against both Gram-negative and Gram-positive bacteria. Furthermore, histones H1.0 and H1.4 demonstrated in vivo efficacy without associated toxicity in an acute infection model of Pseudomonas aeruginosa PAO1 in Galleria mellonella larvae. The bactericidal effect of these proteins also resulted in biomass reduction of P. aeruginosa PAO1 biofilms. Given the clinical significance of this opportunistic pathogen, our research provides a comprehensive initial evaluation of the efficacy, toxicity, and mechanism of action of a potential new therapeutic approach against acute and chronic bacterial infections.

Results of a 5 year Antimicrobial Stewardship program in a North Italian Hospital

Abstract Issue A National Action Plan (PNCAR) to fight antimicrobial resistance has been launched in Italy in 2017, further revised in 2022. In order to reduce inappropriate exposure to antibiotics and infections from multidrug resistant bacteria, a structured system of surveillance and monitoring according to PNCAR standards has been set up in all Italian regions. In the Hospital of Cittadella (District of Padua, Veneto region), special focus has been given to the reduction of antibiotic consumption, in particular quinolones, third generation cephalosporins, carbapenems and macrolides. Description Multidisciplinary meetings have been monthly organized by the hospital Antimicrobial Stewardship Group, with the aim of coordinating the implementation of specific stewardship actions. A quantitative estimate of antibiotic consumption (defined daily dose - DDD - consumption per 100 days of hospitalization) has been provided by the Regional data warehouse from 2019 to 2023 regarding all classes of antibiotics. Results Between 2019 and 2023 a clear decrease in the consumption of quinolones (from 8 to 5,8 DDD, -28%) and carbapenems (from 2,5 to 1,3 DDD, -48%) was observed but, at the same time, an increase in the consumption of third generation cephalosporins (from 9 to 15,7 DDD, +74%) has been reported. This can be explained as a direct consequence of the correct application of therapy guidelines, which in several cases suggest the use of ceftriaxone instead of meropenem. Finally, after an increase in the consumption of macrolides during the first COVID-19 pandemic wave, the consumption of this class of antibiotics has returned to pre-pandemic level. Lessons Constant monitoring of antimicrobial consumption and the identification of warning situations that may need to implement specific actions are the cornerstone of Antimicrobial Stewardship programs. Targets must be defined analyzing data of bacterial resistance rates and infections from multidrug resistant bacteria. Key messages • In Antimicrobial Stewardship programs it is important to adopt constant monitoring of antimicrobial consumption. • Antimicrobial stewardship programs need to be adapted according to the characteristics of each hospital.

Statistical Design and Analysis for Enhanced Production of Tetracycline‐Conjugated Copper Oxide Nanoparticles with Improved Antimicrobial Properties

AbstractPurpose: Since decades, numerous antibiotics have been used to treat different bacterial illnesses; among these, tetracycline is one of the commonly used antibiotics, which has led to development of bacterial resistance. To tackle this critical issue, we have developed a novel, biocompatible nanodrug that improves the antibacterial activity of tetracycline by conjugating it with copper oxide nanoparticles, thereby addressing the problem of multidrug resistance in bacteria. Methods: In this study, using unipot approach, tetracycline conjugated copper oxide nanoparticles were synthesized where tetracycline act as reducing as well as stabilizing agent. These nanoparticles were characterized by different analytical and microscopic technique. For statistical analysis, FFD and CCD model were employed to assess the effect of various parameters (concentration, pH, temperature and time). Results: The synthesis of nanoparticles was confirmed through different techniques. Synthesized nanoparticles exhibited the highest antibacterial activity against various microbes including multi‐drug resistant microbes compared to its individual constituents. DPPH and ABTS assays revealed high antioxidant activity of nanoparticles whereas, biocompatibility was assessed via MTT assay. The drug release profile confirms the sustained release of tetracycline molecules. Conclusions: We have successfully developed tetracycline conjugated copper oxide nanoparticles exhibiting potent antibacterial, antioxidant and biocompatible activity.

The Challenge of Treating Infections Caused by Metallo-β-Lactamase-Producing Gram-Negative Bacteria: A Narrative Review.

Gram-negative multidrug-resistant (MDR) bacteria, including Enterobacterales, Acinetobacter baumannii, and Pseudomonas aeruginosa, pose a significant challenge in clinical practice. Infections caused by metallo-β-lactamase (MBL)-producing Gram-negative organisms, in particular, require careful consideration due to their complexity and varied prevalence, given that the microbiological diagnosis of these pathogens is intricate and compounded by challenges in assessing the efficacy of anti-MBL antimicrobials. We discuss both established and new approaches in the treatment of MBL-producing Gram-negative infections, focusing on 3 strategies: colistin; the recently approved combination of aztreonam with avibactam (or with ceftazidime/avibactam); and cefiderocol. Despite its significant activity against various Gram-negative pathogens, the efficacy of colistin is limited by resistance mechanisms, while nephrotoxicity and acute renal injury call for careful dosing and monitoring in clinical practice. Aztreonam combined with avibactam (or with avibactam/ceftazidime if aztreonam plus avibactam is not available) exhibits potent activity against MBL-producing Gram-negative pathogens. Cefiderocol in monotherapy is effective against a wide range of multidrug-resistant organisms, including MBL producers, and favorable clinical outcomes have been observed in various clinical trials and case series. After examining scientific evidence in the management of infections caused by MBL-producing Gram-negative bacteria, we have developed a comprehensive clinical algorithm to guide therapeutic decision making. We recommend reserving colistin as a last-resort option for MDR Gram-negative infections. Cefiderocol and aztreonam/avibactam represent favorable options against MBL-producing pathogens. In the case of P. aeruginosa with MBL-producing enzymes and with difficult-to-treat resistance, cefiderocol is the preferred option. Further research is needed to optimize treatment strategies and minimize resistance.

Metallic nanoparticles: a promising novel therapeutic tool against antimicrobial resistance and spread of superbugs.

In recent years, antimicrobial resistance (AMR) has become an alarming threat to global health as notable increase in morbidity and mortality has been ascribed to the emergence of superbugs. The increase in microbial resistance because of harboured or inherited resistomes has been complicated by the lack of new and effective antimicrobial agents, as well as misuse and failure of existing ones. These problems have generated severe and growing public health concern, due to high burden of bacterial infections resulting from scarce financial resources and poor functioning health systems, among others. It is therefore, highly pressing to search for novel and more efficacious alternatives for combating the action of these super bacteria and their infection. The application of metallic nanoparticles (MNPs) with their distinctive physical and chemical attributes appears as promising tools in fighting off these deadly superbugs. The simple, inexpensive and eco-friendly model for enhanced biologically inspired MNPs with exceptional antimicrobial effect and diverse mechanisms of action againsts multiple cell components seems to offer the most promising option and said to have enticed many researchers who now show tremendous interest. This synopsis offers critical discussion on application of MNPs as the foremost intervening strategy to curb the menace posed by the spread of superbugs. As such, this review explores how antimicrobial properties of the metallic nanoparticles which demonstrated considerable efficacy against several multi-drugs resistant bacteria, could be adopted as promising approach in subduing the threat of AMR and harvoc resulting from the spread of superbugs.

A Retrospective Closed Cohort Study on Distribution of Multidrug-Resistant Bacteria in Ventilator-Associated Pneumonia and its Impact on Patient Outcome.

Ventilator-associated pneumonia (VAP) is a common and serious nosocomial infection affecting critically ill patients undergoing mechanical ventilation. This study investigated the prevalence of multidrug-resistant (MDR) organisms in VAP, the VAP rate, and the outcomes associated with MDR-VAP. This retrospective single-center study, conducted in 2022, included adult ICU cases from April 2021 to March 2022, receiving mechanical ventilation for more than 48 h. Patient data were analyzed for demographics, comorbidities, empirical antibiotic use, and outcomes. MDR organisms were identified in respiratory cultures. Among 447 patients, 133 developed VAP, with 96 cases being MDR-VAP. The mean age of the overall VAP population was 52 years, 70% of which were males. The incidence of VAP was 30.0% (95% CI: 25.7%-34.5%), while that of MDR-VAP was 21.6% (95% CI: 17.9%-25.8%). The most prevalent MDR organisms were Acinetobacter species (50%) and Klebsiella pneumoniae (46.9%). Empirical antibiotics were administered in 96% of VAP cases. The overall VAP rate was 38.03/1000 ventilator days. No single antimicrobial agent seemed to offer an empirical cover, as the susceptibility rate for most tested antimicrobials was less than 85%. Patients with MDR-VAP had a low survival rate (64.6%) and were less likely to be extubated at 13.5% compared to non-MDR-VAP (survival rate of 62.2%). COVID-19 patients had a high incidence of MDR VAP, especially with Acinetobacter. Overall, VAP mortality was 57.1%. The median ventilator days were 16 for VAP and only four for non-VAP. Gram-negative organisms, particularly Klebsiella and Acinetobacter, were the main MDR VAP culprits. MDR-VAP exhibited higher morbidity and mortality. A study focused on developing resistance by microorganisms is warranted for further understanding.

Antibacterial activity of green synthesized copper oxide nanoparticles against multidrug-resistant bacteria

Using plant extracts in the green synthesis of nanoparticles has become an environmentally acceptable approach. In our study, copper oxide nanoparticles (CuO NPs) were synthesized using ethanolic extracts of Azadirachta indica and Simmondsia chinensis. CuO NP formation was confirmed by the change in color and by UV‒visible spectroscopy (CuO NPs peaked at a wavelength of 344 nm). TEM images confirmed the semispherical shape of the CuO NPs, with particle sizes ranging from 30.9 to 10.7 nm. The antibacterial activity of these NPs was evaluated by using the agar diffusion method against clinical isolates, including methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, Pseudomonas aeruginosa, Acinetobacter spp., Klebsiella pneumoniae, and Stenotrophomonas maltophilia. The minimum inhibitory concentration (MIC) of CuO NPs ranged from 62.5 to 125 µg/ml. In contrast, the antioxidant activity and antibiofilm activity of CuO NPs ranged from 31.1 to 92.2% at 125–500 µg/ml and 62.2–95%, respectively, at 125 –62.5 µg/ml. Our results confirmed that CuO NPs had IC50s of 383.41 ± 3.4 and 402.73 ± 1.86 at 250 µg/mL against the HBF4 cell line. Molecular docking studies with CuO NPs suggested that penicillin-binding protein 4 (PBP4) and beta-lactamase proteins (OXA-48) strongly bind to S. aureus and K. pneumoniae, respectively, with CuO NPs. Our study confirms the promising use of CuO NPs in treating pathogenic bacteria and that CuO NPs could be possible alternative antibiotics. This study supports the pharmaceutical and healthcare sectors in Egypt and worldwide.

Conformational Flexibility and Net Charge are Key Determinants for the Antimicrobial Activity of Peptide Uy234 Against Multidrug-resistant Bacteria

BackgroundThe antimicrobial activity of two peptides, Uy234 derived from the venom of the scorpion Urodacus yaschenkoi and a consensus peptide QnCs-Buap, was evaluated. We tested different pathogenic bacteria: Acinetobacter baumannii, Klebsiella pneumoniae, Salmonella enterica, Bacillus subtilis, Enterococcus spp. and Staphylococcus aureus, including one methicillin resistant (MRSA) and two multidrug resistant (MDR) clinical isolates. In contrast to the QnCs-Buap peptide, Uy234 showed relevant growth inhibitory activity on A. baumannii and B. subtilis, and mostly on S. aureus strains.ObjectiveThe present research focused on elucidating the mechanism for this antibacterial activity.MethodologyWe carried out an in-depth analysis of the composition, structure, flexibility, and physicochemical properties of both peptides.ResultsWe found a crucial role of the C-terminal amide and composition in favoring the formation of a dense H-bond network in the Uy234 peptide. This H-bonding network slightly stiffens the peptide and keeps it in a preordered conformation in the aqueous phase.ConclusionsWe hypothesize that, given that Uy234 is a very short peptide (18 aa), it could have a destabilizing effect and favor micellization phenomena instead forming pores. In contrast, the QnCs-Buap peptide (13 aa), having only the positive charge at the N-terminal end and being significantly more hydrophobic and rigid, is not capable of overcoming the energy barrier to disturb the membrane. We propose that Uy234 peptide can be a scaffold to develop new derivatives with high potential against infections caused by diverse multidrug-resistant bacteria.Graphical

Colistin-niclosamide-loaded nanoemulsions and nanoemulsion gels for effective therapy of colistin-resistant Salmonella infections.

Colistin (COL) is regarded as a last-resort treatment for infections by multidrug-resistant (MDR) Gram-negative bacteria. The emergence of colistin-resistant Enterobacterales poses a significant global public health concern. Our study discovered that niclosamide (NIC) reverses COL resistance in Salmonella via a checkerboard assay. However, poor solubility and bioavailability of NIC pose challenges. In this study, we prepared a self-nanoemulsifying drug delivery system (SNEDDS) co-encapsulating NIC and COL. We characterized the physicochemical properties of the resulting colistin-niclosamide-loaded nanoemulsions (COL/NIC-NEs) and colistin-niclosamide-loaded nanoemulsion gels (COL/NIC-NEGs), assessing their antibacterial efficacy in vitro and in vivo. The COL/NIC-NEs exhibited a droplet size of 19.86 nm with a zeta potential of -1.25 mV. COL/NIC-NEs have excellent stability, significantly enhancing the solubility of NIC while also demonstrating a pronounced sustained-release effect. Antimicrobial assays revealed that the MIC of COL in COL/NIC-NEs was reduced by 16-128 times compared to free COL. Killing kinetics and scanning electron microscopy confirmed enhanced antibacterial activity. Antibacterial mechanism studies reveal that the COL/NIC-NEs and COL/NIC-NEGs could enhance the bactericidal activity by damaging cell membranes, disrupting proton motive force (PMF), inhibiting multidrug efflux pump, and promoting oxidative damage. The therapeutic efficacy of the COL/NIC-NEs and COL/NIC-NEGs is further demonstrated in mouse intraperitoneal infection models with COL-resistant Salmonella. To sum up, COL/NIC-NEs and COL/NIC-NEGs are a potentially effective strategies promising against COL-resistant Salmonella infections.

Plasmonic Supramolecular Nanozyme-Based Bio-Cockleburs for Synergistic Therapy of Infected Diabetic Wounds.

Diabetic wounds are a major devastating complication of diabetes due to hyperglycemia, bacterial invasion, and persistent inflammation, and the current antibiotic treatments can lead to the emergence of multidrug-resistant bacteria. Herein, a bimetallic nanozyme-based biomimetic bio-cocklebur (GNR@CeO2@GNPs) is designed and synthesized for diabetic wound management by depositing spiky ceria (CeO2) shells and gold nanoparticles (GNPs) on a gold nanorod (GNR) nanoantenna. The plasmonic-enhanced nanozyme catalysis and self-cascade reaction properties simultaneously boost the two-step enzyme-mimicking catalytic activity of GNR@CeO2@GNPs, leading to a significant improvement in overall therapeutic efficacy rather than mere additive effects. Under the glucose activation and 808nm laser irradiation, GNR@CeO2@GNPs material captures photons and promotes the transfer of hot electrons from GNR and GNPs into CeO2, realizing a "butterfly effect" of consuming local glucose, overcoming the limited antibacterial efficiency of an individual PTT modality, and providing substantial reactive oxygen species. In vitro and in vivo experiments demonstrate the material's exceptional antibacterial and antibiofilm properties against Gram-negative and Gram-positive bacteria, which can reduce inflammation, promote collagen deposition, and facilitate angiogenesis, thereby accelerating wound healing. This study provides a promising new strategy to develop plasmonic-enhanced nanozymes with a catalytic cascade mode for the antibiotic-free synergistic treatment of infected diabetic wounds.

Nanoscale Multipatterning Zn,Co-ZIF@FeOOH for Eradication of Multidrug-Resistant Bacteria and Antibacterial Treatment of Wounds.

The rising incidence of infections caused by multidrug-resistant bacteria highlights the urgent need for innovative bacterial eradication strategies. Metal ions, such as Zn2+ and Co2+, have bactericidal effects by disrupting bacterial cell membranes and interfering with essential cellular processes. This has led to increased attention toward metal-organic frameworks (MOFs) as potential nonantibiotic bactericidal agents. However, the uniform and enhanced localized release of bactericidal metal ions remains a challenge. Herein, we introduce a nanoscale multipatterned Zn,Co-ZIF@FeOOH, featuring a multipod-like morphology with spiky corners, and dual-bactericidal metal ions. Compared to pure Zn,Co-ZIF, the multipod-like morphology of Zn,Co-ZIF@FeOOH exhibits enhanced adhesion toward bacterial surfaces via topological and multiple interactions of electrostatic interaction, significantly increasing the local release of Zn2+ and Co2+. Additionally, the spiky corners of the spindle-shaped FeOOH nanorods physically penetrate bacterial membranes, causing damage and further enhancing adhesion to bacteria. Nine Gram-negative and one Gram-positive bacteria were selected for in vitro test. Notably, the nanoscale multipatterned Zn,Co-ZIF@FeOOH exhibited high bactericidal efficacy against various multidrug-resistant bacteria, including extended-spectrum β-lactamase-producing (ESBL+) bacteria and carbapenem-resistant bacteria, performing well in both acidic and neutral environments. The wound healing activity of Zn,Co-ZIF@FeOOH was further demonstrated using female Balb/c mouse models infected with bacteria, where the materials show robust antibacterial efficacy and commendable biocompatibility. This study showcases the assembly of metal oxide/MOF composites for nanoscale multipatterning, aims at synergistic bacterial eradication and offers insights into developing nanomaterial-based strategies against multidrug-resistant bacteria.

Metapopulation model of phage therapy of an acute Pseudomonas aeruginosa lung infection.

Phage therapy is increasingly employed as a compassionate treatment for severe infections caused by multidrug-resistant (MDR) bacteria. However, the mixed outcomes observed in larger clinical studies highlight a gap in understanding when phage therapy succeeds or fails. Previous research from our team, using in vivo experiments and single-compartment mathematical models, demonstrated the synergistic clearance of acute P. aeruginosa pneumonia by phage and neutrophils despite the emergence of phage-resistant bacteria. In fact, the lung environment is highly structured, prompting the question of whether immunophage synergy explains the curative treatment of P. aeruginosa when incorporating realistic physical connectivity. To address this, we developed a metapopulation network model mimicking the lung branching structure to assess phage therapy efficacy for MDR P. aeruginosa pneumonia. The model predicts the synergistic elimination of P. aeruginosa by phage and neutrophils but emphasizes potential challenges in spatially structured environments, suggesting that higher innate immune levels may be required for successful bacterial clearance. Model simulations reveal a spatial pattern in pathogen clearance where P. aeruginosa are cleared faster at distal nodes of the bronchial tree than in primary nodes. Interestingly, image analysis of infected mice reveals a concordant and statistically significant pattern: infection intensity clears in the bottom before the top of the lungs. The combined use of modeling and image analysis supports the application of phage therapy for acute P. aeruginosa pneumonia while emphasizing potential challenges to curative success in spatially structured in vivo environments, including impaired innate immune responses and reduced phage efficacy.