Resistance Crisis Research Articles

Polyelectrolytes and Polyelectrolyte Complexes as Future Antibacterial Agents

Antibiotic-resistance (ABR) poses a critical public health challenge within the broader antimicrobial resistance crisis. This review evaluates the potential of polyelectrolytes (PEs) and polyelectrolyte complexes (PECs) for controlled antibiotic delivery as a strategy to combat ABR and biofilm-related infections. PECs, particularly those incorporating chitosan and other polycations, enhance antibacterial efficacy by disrupting bacterial cell walls and obstructing their nutrient flow. They are also effective in penetrating biofilms and providing sustained drug release. Despite these advantages, there is a need for further in vivo research and clinical trials to validate these findings. This review provides a comprehensive overview of PECs’ potential to advance antibacterial therapies and outlines future research directions to further explore their applications.

Antibiotic Resistance in humans due to Microbes

Background Antibiotics are now facing a crisis of resistance. Overuse and misuse have led to bacteria developing resistance, making infections harder to treat. This is exacerbated by the availability of antibiotics without prescriptions and their presence in the environment. The spread of resistant bacteria poses a significant threat to global health, requiring urgent action. Governments and international organizations are working to combat this issue, but the problem persists. Addressing antibiotic resistance requires responsible use and the development of new antibiotics. Objective This study aimed to review antibiotic resistance in humans due to microbes. Discussion Antimicrobial resistance (AMR) is a natural process where microorganisms stop responding to antibiotics, making infections harder to treat and increasing the risk of spreading serious diseases. The overuse of antibiotics accelerates this process, promoting AMR spread through distinct mechanisms like natural and acquired resistance. Genetic mechanisms involve inherent and acquired resistance in bacteria, impacting antibiotic effectiveness. Biological resistance mechanisms include enzymatic inhibition, reduced penetration, enhanced extrusion, and target changes. Strategies against antimicrobial agents involve restricting drug entry, modifying targets, rendering drugs inactive, and actively removing drugs from cells. Gram-negative and gram-positive bacteria exhibit variations in resistance mechanisms due to structural differences. Conclusion The development of antibiotic resistance in bacteria is a multifaceted issue, involving both inherent and acquired mechanisms. Inherent resistance stems from genetic mutations that occur spontaneously within bacterial populations. On the other hand, acquired resistance can occur through the transfer of genetic material between bacteria, enabling the spread of resistance traits. It is imperative to grasp these genetic, biological, and strategic mechanisms to effectively tackle the escalating challenge of antibiotic resistance and devise successful strategies to mitigate its impact.

Multiomics comparative analysis of feces AMRGs of Duroc pigs and Tibetan and the effect of fecal microbiota transplantation on AMRGs upon antibiotic exposure.

To the best of our knowledge, this study represents the first comprehensive analysis of antimicrobial resistance gene (AMRGs) expression in the fecal microbiota of Tibetan and Duroc pigs, employing an integrated metagenomic and metatranscriptomic approach. Our findings indicate a higher risk of AMRGs transmission in the feces of Duroc pigs compared to Tibetan pigs. Given the escalating antimicrobial resistance crisis, novel therapeutic interventions are imperative to mitigate gut colonization by pathogens and AMRGs. In this regard, we investigated the impact of fecal microbiota from Tibetan and Duroc pig sources on AMRGs excretion in Duroc Landrace Yorkshire (DLY) piglets' feces following acute antibiotic exposure. Remarkably, only fecal microbiota sourced from Tibetan pigs exhibited a reduction in AMRGs excretion in DLY piglets' feces. This underscores the significance of evaluating the presence of AMRGs within donor fecal microbiota for effective AMRGs decolonization strategies.

National Multicenter Study on the Prevalence of Carbapenemase-Producing Enterobacteriaceae in the Post-COVID-19 Era in Argentina: The RECAPT-AR Study

The COVID-19 pandemic has exacerbated the global antimicrobial resistance (AMR) crisis. Consequently, it is more urgent than ever to prioritize AMR containment and support countries in improving the detection, characterization, and rapid response to emerging AMR threats. We conducted a prospective, multicenter study to assess the prevalence of carbapenemase-producing Enterobacterales in infectious processes in Argentina during the post-COVID-19 pandemic period and explore therapeutic alternatives for their treatment (RECAPT-AR study). Methods: A total of 182 hospitals participated by submitting Enterobacterales clinical isolates to the National Reference Laboratory (NRL) during the first three weeks of November 2021. Inclusion criteria were defined as an ertapenem MIC ≥ 0.5 mg/L, a zone diameter ≤ 22 mm. Carbapenemase genes and those coding for major extended-spectrum β-lactamases were molecularly characterized using multiplex PCR at the NRL. Antibiotic susceptibility testing followed international standards (CLSI and EUCAST). Results: The NRL analyzed 821 Enterobacterales isolates. Metallo-β-lactamase (MBL, 42.0%) and KPC (39.8%) accounted for 81.8% of carbapenemases, followed by OXA-163 (7.4%), a variant of OXA-48 with additional activity against extended-spectrum cephalosporins, and enzyme combinations (8.3%). These combinations included NDM plus KPC (3.4%), OXA-163 plus KPC (2.4%), and OXA-163 plus NDM (2.1%). Klebsiella pneumoniae was the main species recovered, representing 76% of the isolates. According to the carbapenemase classes or combinations, tigecycline exhibited a susceptibility range of 33–83%, fosfomycin 59–81%, colistin 27–78%, and amikacin 17–81%. Ceftazidime-avibactam (CZA) and imipenem-relebactam (IMR) showed 92% and 98% susceptibility against serine carbapenemases, respectively. Meanwhile, aztreonam-avibactam (AZA) exhibited 96–98% susceptibility against all carbapenemase classes. Conclusions: A new epidemiological landscape has emerged, characterized by the equivalent circulation of NDM and KPC. K. pneumoniae remains the primary species responsible for their dissemination. The co-production of carbapenemase combinations, particularly KPC plus NDM, was confirmed, mainly in K. pneumoniae. High activity was observed for AZA against MBLs and for CZA and IMR against KPC and OXA-163 carbapenemases.

Synergistic antimicrobial activity of lynronne-1 and EDTA against bovine mastitis pathogens.

Bovine mastitis is the costliest disease in the dairy sector and the main cause of antibiotic use in dairy cattle, potentially contributing to the antimicrobial resistance crisis. Antimicrobial peptides (AMPs) offer promise as antibiotic alternatives for controlling mastitis pathogens. The efficacy of five AMPs (Lynronne-1 [Lyn-1], Lynronne-2 [Lyn-2], Bovicin HC5, AMP 660, and AMP 1043) and two bioactive compounds (disodium ethylenediaminetetraacetic acid [EDTA] and glycerol monolaurate) was assessed against a range of 35 mastitis-causing pathogens. The fractional inhibitory concentrations index (FICI) was calculated to determine the interaction effect and values ≤0.5 were indicative of synergism. Time-dependent killing assays were performed to assess bactericidal efficacy of the combination. Cytotoxicity was evaluated using the MTT assay and haemolytic activity was assessed against fresh bovine erythrocytes. Lyn-1 and EDTA exhibited the highest broad spectrum antimicrobial activity and reduced bacterial growth (OD600 nm) by 95.1% and 86.9%, respectively. FICI values ranged from 0.1 to 0.5, indicating synergism. The combination of lyn-1 (0.03 mg/mL) and EDTA (1.02 mg/mL) exhibited higher antimicrobial activity against all bacterial strains, at significantly lower concentrations than each compound individually. Lyn-1-EDTA combination reduced viable population by >10 000-fold within 12 h. The combination was non-haemolytic in concentrations up to 8-fold the established MIC values (P > 0.05), although cytotoxic effects were observed at concentrations above MIC (P < 0.01). These findings highlight the therapeutic potential of Lyn-1 and Lyn-1-EDTA for developing antibiotic-free formulations to combat contagious and environmental mastitis pathogens and treat udder infections.

Characterization and Comparative Genomic Analysis of vB_BceM_CEP1: A Novel Temperate Bacteriophage Infecting Burkholderia cepacia Complex.

The increasing prevalence of multidrug-resistant bacteria imminently threatens public health and jeopardizes nearly all aspects of modern medicine. The Burkholderia cepacia complex (Bcc) comprises Burkholderia cepacia and the related species of Gram-negative bacteria. Members of the Bcc group are opportunistic pathogens responsible for various chronic illnesses, including cystic fibrosis and chronic granulomatous disease. Phage therapy is emerging as a potential solution to combat the antimicrobial resistance crisis. In this study, a temperate phage vB_BceM_CEP1 was isolated from sewage and fully characterized. Transmission electron microscopy indicated that vB_BceM_CEP1 belongs to the family Peduoviridae. The isolated phage demonstrated enhanced environmental stability and antibiofilm potential. One-step growth analysis revealed a latent period of 30min and an average burst size of 139 plaque-forming units per cell. The genome of vB_BceM_CEP1 consists of 32,486bp with a GC content of 62.05%. A total of 40 open reading frames were annotated in the phage genome, and none of the predicted genes was annotated as tRNA. Notably, genes associated with antibiotic resistance, host virulence factors, and toxins were absent from the vB_BceM_CEP1 genome. Based on its unique phenotype and phylogeny, the isolated phage vB_BceM_CEP1 is classified as a new temperate phage with lytic activity. The findings of this study enhance our understanding of the diversity of Bcc phages.

FAGOTERAPIA COMO ALTERNATIVA NO COMBATE A INFECÇÕES RESISTENTES: AVANÇOS E DESAFIOS NO TRATAMENTO DE E. COLI UPEC E EPEC

Resistant bacterial infections represent one of the biggest public health challenges today, resulting in a significant increase in morbidity and mortality linked to infectious diseases. On the other hand, Escherichia coli stands out as one of the most significant bacteria, particularly the uropathogenic (UPEC) and enteropathogenic (EPEC) variants. Infections caused by these strains have become a problem due to their ability to develop resistance to various antibiotics, thus harming therapeutic alternatives. The objective of this research was to investigate the effectiveness of phage therapy as an alternative in combating resistant bacterial infections, focusing on UPEC and enteropathogenic EPEC strains of Escherichia coli. Methodologically, it was an exploratory and comparative bibliographic review through qualitative research. As a result, it was found that future directions in phage therapy present a promising outlook, full of technological innovations, research needs and ethical considerations. The success of phage therapy as a viable alternative to treating resistant bacterial infections will depend on collaboration between different disciplines, careful consideration of ethical implications, and a commitment to ensuring that new technologies benefit society as a whole. The development of robust guidelines and regulations, along with increased public awareness, will be critical to the effective integration of phage therapy into clinical practice and public health. It was concluded that progress in this area represents a response to the bacterial resistance crisis, as it opens up new perspectives for biomedical research and clinical practice. Therefore, the continued engagement of researchers, health professionals and policy makers will be essential to ensure that phage therapy is established as an integral component in the fight against bacterial infections, benefiting public health in a broad and effective way.

Antibiofilm carbon-wrapped copper-nickel nanoparticles loaded amorphous hydrogel accelerated healing of Pseudomonas aeruginosa infected wounds in rat model

Pseudomonas aeruginosa induced infections are becoming very common and bacterial resistance to antibiotics causes a significant challenge to health systems globally by imposing massive financial burdens. Finding antibiotics substitutes with better antibacterial activity and lower risk are highly desired. Metallic nanoparticles have been found to be very attractive antibacterial agents. In current research, Carbon-wrapped Copper-Nickel nanoparticles were prepared and characterized using Fourier Transform Infrared spectroscopy (FT-IR) and X-ray diffraction analysis. Propylene glycol, sodium alginate, and 2-phenoxy ethanol-based hydrogels were loaded with two different concentrations of Copper-Nickel nanoparticles 30 mg/ml (ALG-30) and 50 mg/ml (ALG-50). The synthesized gels were found to be cytocompatible in cell culture study using fibroblasts. Carbon-wrapped Copper-Nickel nanoparticles, ALG-30 and ALG-50 showed remarkable antibacterial activity against both Gram-positive (S. aureus and MRSA) and Gram-negative (E. coli and P. aeruginosa) bacteria. Pseudomonas aeruginosa and MRSA biofilms were significantly decreased when exposed to nanoparticles and hydrogels (ALG-30 and ALG-50) for 24 hours. In full thickness infected skin wound model in rat, Pseudomonas aeruginosa infected wound was almost 97 % healed with ALG-30 over 16 days. In ALG-30 treated group, H & E stained tissues depicted enhanced angiogenesis, reduced inflammation and stimulate re-epithelialization, while Masson trichrome stained tissues showed increased collagen deposition. Overall, Carbon-wrapped Copper-Nickel nanoparticles embedded hydrogels exhibited excellent antimicrobial activity and holds a considerable potential to alleviate the current antibiotic resistance crisis in chronically infected wounds and expected widespread use in clinical practice.

Uncovering the arsenal of class II bacteriocins in salivarius streptococci

Facing the antibiotic resistance crisis, bacteriocins are considered as a promising alternative to treat bacterial infections. In the human commensal Streptococcus salivarius, the production of unmodified bacteriocins (or salivaricins) is directly controlled at the transcriptional level by quorum-sensing. To discover hidden bacteriocins, we harnessed here the unique molecular signatures of salivaricins not yet used in available computational pipelines and performed genome mining followed by orthogonal reconstitution and expression. From 100 genomes of S. salivarius, we identified more than 50 bacteriocin candidates clustered into 21 groups. Strain-based analysis of bacteriocin combinations revealed significant diversity, reflecting the plasticity of seven independent loci. Activity tests showed both narrow and broad-spectrum bacteriocins with overlapping activities against a wide panel of Gram-positive bacteria, including notorious multidrug-resistant pathogens. Overall, this work provides a search-to-test generic pipeline for bacteriocin discovery with high impact for bacterial ecology and broad applications in the food and biomedical fields.

Biofilm targeting with chitosan-based nanohydrogel containing Quercus infectoria G. Olivier extract against Streptococcus mutans: new formulations of a traditional natural product

BackgroundBiofilm formation has a crucial role in the cariogenic virulence of Streptococcus mutans, which leads to resistance to common antibacterials. The antimicrobial resistance crisis has led to increased research about traditional natural products.PurposeQuercus infectoria extract (QI extract) and nano hydrogels containing QI extract (QI-NH) and tannic acid (TA-NH) were evaluated against this pathogen.MethodsQI extract was analyzed by HPLC and the physiological characteristics of nanohydrogels were assessed by SEM, FTIR, zeta potential, DLS and determination of release kinetics and encapsulation efficiency. Determination of MIC and MBC of the material and their anti-biofilm effect was done by the microtiter method and on the extracted tooth surface. The properties of extracts and nano hydrogels in the expression of genes codifying glucosyltransferases (gtfB, gtfC and gtfD) and glucan binding protein B (gbpB) were quantified. Their toxicity was tested by the MTT method against the KB cell line.ResultsAccording to HPLC, 55.18% of QI extract contained TA. The encapsulation efficiency of QI-NH and TA-NH was equal to 60% and 80%, respectively. SEM and FTIR exhibited that QI extract and TA were successfully entrapped in the networks resulting from the chemical bonding of chitosan and TPP. The average size of QI-NH and TA-NH was 70.45 and 58.43 nm, and their zeta potential was 6.17 ± 2.58 and 0.25 ± 0.03 mv, respectively. PDI < 0.3 of nano hydrogels indicated the favorable polydispersity of nanohydrogels. MIC of QI extract, QI-NH and TA-NH were 937.5, 30 and 10 µg/ml, respectively. Also their MBIC50 was 35.1, 2.1 and 0.95 µg/ml, respectively, and the extracts and nano hydrogels restrained the biofilm maturation on enamel. The pivotal genes of S. mutans in biofilm formation were significantly less expressed by treatment with QI-NH and TA-NH than others. Based on the MTT test, QI-NH had less acute toxicity than QI extract and TA-NH. IC50 of QI-NH was calculated as 775.4 µg/ml, while it was equal to 3.12 µg/ml for chlorhexidine as a common antibacterial agent.ConclusionQI-NH, a new formulation derived from traditional anti-caries, can be a safe and efficient option to combat dental biofilm.

ACTINOBACTERIAL DIVERSITY DETERMINATION IN MALAYSIAN BRIS (BEACH RIDGES INTERSPERSED WITH SWALES) SOIL BY CULTURE-DEPENDENT APPROACH

Globally, the rapid emergence of antibiotic-resistant bacteria poses a major threat to public health, threatening the effectiveness of antibiotics. Addressing the crisis of antimicrobial resistance requires urgency in new drug discovery and the effort in finding novel compounds is dramatically evolving. However, one of the main obstacles is the rediscovery of known compounds from natural resources. To overcome this challenge, exploring underexploited location for new resource of actinomycetes, such as Beach Ridges Interspersed with Swales (BRIS) soil in Malaysia, was selected. Soil samples from Jambu Series and Rudua Series in Setiu, Terengganu were collected and analysed. A total of 87 actinomycetes strains were isolated on yeast malt agar (ISP2), Actinomyces agar and Zobell marine agar, with the highest number from the Jambu series using serial dilution and spread plate method. Antimicrobial screening revealed that 31 isolates exhibited activity against ESKAPE pathogens, with 28 and 12 isolates showing activity against Gram-positive and Gram-negative bacteria, respectively. These isolates showed strong inhibiting response towards one or more ESKAPE pathogens. This study highlights the potential of BRIS soil as an underexploited habitat for discovering actinobacterial communities which could lead to the discovery of a new potential antimicrobial producer to serve as novel drug leads.

Bacteria carrying mobile colistin resistance genes and their control measures, an updated review.

The plasmid encoded mobile colistin resistance (MCRs) enzyme poses a significant challenge to the clinical efficacy of colistin, which is frequently employed as a last resort antibiotic for treating infections caused by multidrug resistant bacteria. This transferase catalyzes the addition of positively charged phosphoethanolamine to lipid A of the outer membrane of gram-negative bacteria, thereby facilitating the acquired colistin resistance. This review aims to summarize and critically discuss recent advancements in the distribution and pathogenesis of mcr-positive bacteria, as well as the various control measures available for treating these infections. In addition, the ecology of mcr genes, colistin-resistance mechanism, co-existence with other antibiotic resistant genes, and their impact on clinical treatment are also analyzed to address the colistin resistance crisis. These insights provide a comprehensive perspective on MCRs and serve as a valuable reference for future therapeutic approaches to effectively combat mcr-positive bacterial infections.

Discovery of a family of menaquinone-targeting cyclic lipodepsipeptides for multidrug-resistant Gram-positive pathogens

Menaquinone (MK) in bacterial membrane is an attractive target for the development of novel therapeutic agents. Mining the untapped chemical diversity encoded by Gram-negative bacteria presents an opportunity to identify additional MK-binding antibiotics (MBAs). By MK-binding motif searching of bioinformatically predicted linear non-ribosomal peptides from 14,298 sequenced genomes of 45 underexplored Gram-negative bacterial genera, here we identify a novel MBA structural family, including silvmeb and pseudomeb, using structure prediction-guided chemical synthesis. Both MBAs show rapid bacteriolysis by MK-dependent membrane depolarization to achieve their potent activities against a panel of Gram-positive pathogens. Furthermore, both MBAs are proven to be effective against methicillin-resistant Staphylococcus aureus in a murine peritonitis-sepsis model. Our findings suggest that MBAs are a kind of structurally diverse and still underexplored antibacterial lipodepsipeptide class. The interrogation of underexplored bacterial taxa using synthetic bioinformatic natural product methods is an appealing strategy for discovering novel biomedically relevant agents to confront the crisis of antimicrobial resistance.

Intersecting social and environmental determinants of multidrug-resistant urinary tract infections in East Africa beyond antibiotic use

The global health crisis of antibacterial resistance (ABR) poses a particular threat in low-resource settings like East Africa. Interventions for ABR typically target antibiotic use, overlooking the wider set of factors which drive vulnerability and behaviours. In this cross-sectional study, we investigated the joint contribution of behavioural, environmental, socioeconomic, and demographic factors associated with higher risk of multi-drug resistant urinary tract infections (MDR UTIs) in Kenya, Tanzania, and Uganda. We sampled outpatients with UTI symptoms in healthcare facilities and linked their microbiology data with patient, household and community level data. Using bivariate statistics and Bayesian profile regression on a sample of 1610 individuals, we show that individuals with higher risk of MDR UTIs were more likely to have compound and interrelated social and environmental disadvantages: they were on average older, with lower education, had more chronic illness, lived in resource-deprived households, more likely to have contact with animals, and human or animal waste. This suggests that interventions to tackle ABR need to take account of intersectional socio-environmental disadvantage as a priority.

Global emergence of Escherichia coli with PBP3 insertions.

Escherichia coli producing metallo-β-lactamases with penicillin-binding protein 3 (PBP3) insertions have reduced susceptibility to aztreonam-avibactam and cefiderocol. Here, we analysed high-quality E. coli genomes for PBP3 insertions. E. coli genomes (n = 167 518) were retrieved from EnteroBase with CheckM2 for quality control, fastANI for species confirmation, multi-locus sequencing typing for sequence type (ST) determination and AMRFinderPlus for resistance gene identification. For PBP3 insertion analysis, we used Prokka for predicting coding sequences, BLAST+ for comparing resulted protein sequences and SnpEff for annotating variants. Among the included 159 341 genomes, PBP3 insertions with 11 variants were found in 2.01% (n = 3198). The predominant variant is a duplication of 334-337 amino acids (aa) (94.75%, n = 3030), comprising YRIN (65.92%, n = 2108) and its single-aa-variant YRIK (28.83%, n = 922), followed by a similar PYRI duplication of 333-336 (4.16%, n = 133). The less common ones are a TIPY duplication of 331-334 (n = 24) and its single-aa-variant TVPY's duplication: TVVPY (n = 1), TVPYTVPY (n = 1) and TVPYPVPY (n = 1). Rare duplications include VGDR of 106-109 (n = 3), ANALNIPL of 114-121 (n = 3), AL of 567-568 (n = 1) and TG of 584-585 (n = 1). Insertion variants were detected across 62 countries on six continents, primarily in human samples, and associated with 85 STs, concentrated in high-risk clones ST410 (29.18%, n = 1923), ST167 (23.40%, n = 1740) and ST405 (10.56%, n = 1334), with 83.32% (n = 2218) encoding metallo-β-lactamase NDM. Global spread of E. coli harbouring PBP3 insertion, often with NDM β-lactamase, high-risk ST410, ST167 and ST405 clones and various hosts, underscores the escalating antimicrobial resistance crisis and the urgency for a 'One Health' strategy.

VFDB 2025: an integrated resource for exploring anti-virulence compounds.

With the escalating crisis of bacterial multidrug resistance, anti-virulence therapeutic strategies have emerged as a highly promising alternative to conventional antibiotic treatments. Anti-virulence compounds are specifically designed to target virulence factors (VFs), disarming pathogens without affecting bacterial growth and thus reduce the selective pressure for resistance development. However, due to the complexity of bacterial pathogenesis, no anti-virulence small molecules have been approved for clinical use thus far, despite the documentation of hundreds of potential candidates. To provide valuable reference resources for drug design, repurposing, and target selection, the virulence factor database (VFDB, http://www.mgc.ac.cn/VFs/) has systematically collected public data on anti-virulence compounds through extensive literature mining, and further integrated this information with its existing knowledge of bacterial VFs. To date, the VFDB has curated a comprehensive dataset of 902 anti-virulence compounds across 17 superclasses reported by 262 studies worldwide. By cross-linking the current knowledge of bacterial VFs with information on relevant compounds (e.g. classification, chemical structure, molecular targets and mechanisms of action), the VFDB aims to bridge the gap between chemists and microbiologists, providing crucial insights for the development of innovative and effective antibacterial therapies to combat bacterial infections and address antibiotic resistance.

Recent Developments in Infectious Disease Research

The field of infectious disease research is rapidly evolving and advancing, driven by the demand to address both current and emerging threats to global health. The recent COVID-19 pandemic underscored the criticality of infectious disease outbreak preparedness and response, while factors such as climate change, migration, and globalization continue to influence disease transmission dynamics. Innovative approaches to infectious disease research are more essential than ever to prevent future pandemics and the spread of vectorborne diseases. This comprehensive review article investigates some recent pertinent developments in infectious disease research, including the development of a novel serosurveillance platform, modeled after the one designed during the COVID-19 pandemic, aimed at tackling the growing global threat of arboviruses. Also investigated in this article is the repurposing and application of mRNA vaccine technology, initially developed during the pandemic, to address the complex challenge of tick-borne Lyme disease. Finally, we outline several novel and cutting-edge strategies aimed at tackling the ever-mounting crisis of antimicrobial resistance (AMR). Each of these innovations marks a significant step forward in global health security and preparedness, offering promising solutions to the challenges of a rapidly evolving world, while drawing on the lessons learned from past outbreaks and epidemics.

Restoring Colistin Sensitivity in Multidrug-Resistant Pathogenic E. coli Using Cinacalcet Hydrochloride.

Restoring colistin's efficacy is crucial in addressing the resistance crisis of colistin. This study utilized a high-throughput screening method to identify 43 compounds from 800 FDA-approved drugs that exhibited significant antibacterial effects when combined with colistin. Among these, cinacalcet hydrochloride (CH) was selected for its potential synergistic effect with colistin against multidrug-resistant (MDR) E. coli strains, including mcr-1-positive strains. A series of experiments revealed that the combination of CH and colistin showed strong synergy, especially in mcr-1-positive strains, restoring colistin sensitivity. The combination significantly inhibited bacterial growth and reduced CFU counts more effectively than either drug alone. Additionally, CH and colistin together significantly inhibited biofilm formation and eradicated existing biofilms, as visualized through confocal microscopy. Mechanistic studies showed that the combination increased bacterial membrane permeability and disrupted membrane integrity. The treatment also elevated extracellular ATP release and ROS production, indicating oxidative stress-induced bacterial death. Safety evaluations showed that the combination did not increase toxicity in host cells. Finally, animal models further validated the combination's efficacy. Overall, this study showed that the combination of colistin and CH significantly restored colistin sensitivity in mcr-1-positive E. coli, revealing their synergistic antibacterial mechanism involving membrane damage and oxidative stress, with promising clinical applications.

The treatment of Enterococcus faecalis related root canal biofilms with phage therapy

As one of the global concerns, antimicrobial resistance crisis increases the clinical importance of Enterococcus species. Enterococcus faecalis (E. faecalis) specifically penetrates the dentinal tubules and remains prevalent even after endodontic treatment. It has also biofilm forming character as well as the development of resistance to antibiotics. Sodium hypochlorite (NaOCl) is considered the gold standard among antibacterial washing solutions. However, due to its toxic effects, its usage limitations have led to the search for natural, non-toxic alternatives. Phages can be considered an important alternative because of their effects on specific bacteria. The objective of this study was to compare the effect of isolated active vB_Ef1 phage on the removal of E. faecalis biofilm in dentin, together and separately with the chemical irrigation solution NaOCl. As a result of study, the optimal NaOCl solution concentration to be applied with vB_Ef1 phage is 0.5 %, and the use of solution at this value reduces the biofilm mass by 84 %, reaching the highest biofilm mass reduction value. It was found that the combination of phage and NaOCl at appropriate concentrations had the strongest biofilm disruption effect.

Plasma activated water pre-treatment substantially enhances phage activity against Proteus mirabilis biofilms

The ongoing antimicrobial resistance crisis has incentivised research into alternative antibacterial and antibiofilm agents. One of them is plasma-activated water (PAW), which is produced by exposing water to a cold plasma discharge. This process generates a diverse array of reactive oxygen and nitrogen species (ROS/RNS) with antimicrobial properties. Another intensively studied class of alternative antimicrobials are bacteriophages, attracting attention due to their specificity and strong antibacterial activity. As combinations of different types of antimicrobials are known to often exhibit synergistic interactions, in this study we investigated the combined use of cold atmospheric-pressure plasma-activated water and the bacteriophage vB_PmiS_PM-CJR against Proteus mirabilis biofilms as a potential option for treatment of catheter-associated urinary tract infections (CAUTIs).We compared the effect of two cold plasma discharge setups for PAW production on its antimicrobial efficacy against P. mirabilis planktonic and biofilm cultures. Next, we assessed the stability of the phage vB_PmiS_PM-CJR in PAW. Finally, we tested the antimicrobial activity of the phages and PAW against biofilms, both individually and in combinations.Our findings demonstrate that the combination of PAW with phage is more effective against biofilms compared to individual treatments, being able to reduce the number of biofilm-embedded cells by approximately 4 log. We were also able to show that the order of treatment plays an important role in the anti-biofilm activity of the phage-PAW combination, as the exposure of the biofilm to PAW prior to phage administration results in a stronger effect than the reverse order.This research underlines PAW's ability to potentiate phage activity, showcasing a considerable reduction in biofilm viability and biomass. Additionally, it contributes to the growing body of evidence supporting the use of phage-based combinatorial treatments. Overall, this sequential treatment strategy demonstrates the potential of leveraging multiple approaches to address the mounting challenge of antibiotic resistance and offers a promising avenue for enhancing the efficacy of CAUTI management.