Antibiotic-resistant Pathogens Research Articles

Redox-Modulating Capacity and Effect of Ethyl Acetate Roots and Aerial Parts Extracts from Geum urbanum L. on the Phenotype Inhibition of the Pseudomonas aeruginosa Las/RhI Quorum Sensing System

Pseudomonas aeruginosa is an opportunistic pathogen that causes nosocomial infections of the urinary tract, upper respiratory tract, gastrointestinal tract, central nervous system, etc. It is possible to develop bacteremia and sepsis in immunocompromised patients. A major problem in treatment is the development of antibiotic resistance. Therefore, new preparations of natural origin are sought, such as plant extracts, which are phytocomplexes and to which it is practically impossible to develop resistance. Geum urbanum L. (Rosacea) is a perennial herb known for many biological properties. This study aimed to investigate the redox-modulating capacity and effect of ethyl acetate (EtOAc) extracts from roots (EtOAcR) and aerial parts (EtOAcAP) of the Bulgarian plant on the phenotype inhibition of the P. aeruginosa Las/RhI quorum sensing (QS) system, which primarily determines drug resistance in pathogenic bacteria, including biofilm formation, motility, and pigment production. We performed QS assays to account for the effects of the two EtOAc extracts. At sub-minimal inhibitory concentrations (sub-MICs) ranging from 1.56 to 6.25 mg/mL, the biofilm formation was inhibited 85% and 84% by EtOAcR and 62% and 39% by EtOAcAP extracts, respectively. At the same sub-MICs, the pyocyanin synthesis was inhibited by 17–27% after treatment with EtOAcAP and 26–30% with EtOAcR extracts. The motility was fully inhibited at 3.12 mg/mL and 6.25 mg/mL (sub-MICs). We investigated the inhibitory potential of lasI, lasR, rhiI, and rhiR gene expression in biofilm and pyocyanin probes with the PCR method. Interestingly, the genes were inhibited by two extracts at 3.12 mg/mL and 6.25 mg/mL. Antiradical studies, assessed by DPPH, CUPRAC, and ABTS radical scavenging methods and superoxide anion inhibition showed that EtOAcAP extract has effective antioxidant capacity. These results could help in the development of new phytocomplexes that could be applied as biocontrol agents to inhibit the phenotype of the P. aeruginosa QS system and other antibiotic-resistant pathogens.

Strain phylogroup and environmental constraints shape Escherichia coli dynamics and diversity over a 20-year human gut time series.

Escherichia coli is an increasingly antibiotic-resistant opportunistic pathogen. Few data are available on its ecological and evolutionary dynamics in its primary commensal niche, the vertebrate gut. Using Illumina and/or Nanopore technologies, we sequenced whole genomes of 210 E. coli isolates from 22 stools sampled during a 20-year period from a healthy man (ED) living in Paris, France. All phylogroups, except C, were represented, with a predominance of B2 (34.3%), followed by A and F (19% each) phylogroups. Thirty-five clones were identified based on their haplogroup and pairwise genomic single nucleotide polymorphism distance and classified in three phenotypes according to their abundance and residence time: 25 sub-dominant/transient (52 isolates), five dominant/transient (48 isolates) and five dominant/resident (110 isolates). Four over five dominant/resident clones belonged to B2 and closely related F phylogroups, whereas sub-dominant/transient clones belonged mainly to B1, A and D phylogroups. The long residence times of B2 clones seemed to be counterbalanced by lower colonization abilities. Clones with larger within-host frequency persisted for longer. By comparing ED strain genomes to a collection of commensal E. coli genomes from 359 French individuals, we identified ED-specific genomic properties including an enrichment in genes involved in a metabolic pathway (mhp cluster) and the presence of a very rare antiviral defense island. The E. coli colonization within the gut microbiota was shaped by both the intrinsic properties of the strain lineages, in particular longer residence of phylogroup B2, and the environmental constraints such as diet or phages.

Antimicrobial and Antibiofilm Activities of the Essential Oil from the Inflorescence of Etlingera elatior (Zingiberaceae) and Its Main Constituents (Dodecanal and 1-Dodecanol)

The spread of antibiotic-resistant pathogens had motivated the search for plant-derived antimicrobial agents, such as essential oils. This work investigated the antibacterial, antifungal and antibiofilm properties of the essential oil from the inflorescence of Etlingera elatior and its major compounds. Dodecanal (42.46%) and 1-dodecanol (25.84%) were the major compounds. The oil, dodecanal and 1-dodecanol were active against Staphylococcus aureus, with minimum bactericidal concentrations of 124, 31 and 62 µg mL-1, respectively. For Pseudomonas aeruginosa, the essential oil had an inhibitory effect against a multidrug resistant isolate (minimum inhibitory concentration (MIC): 4,000 µg mL-1). Dodecanal had an MIC of 60,000 µg mL-1 for all Candida species, but fungicidal activity was only detected for Candida albicans (minimum fungicidal concentration: 60,000 µg mL-1). 1-Dodecanol (60,000 µg mL-1) exhibited fungistatic activity for C. albicans. Biofilm inhibition linked to an anti-adhesive effect of the oil and dodecanal was only detected for the standard S. aureus isolate and C. albicans, respectively. In conclusion, the essential oil from the inflorescence of E. elatior and its major components were more active against nonresistant strains S. aureus. Essential oil, dodecanol and dodecanal showed excellent activity against S. aureus, however, these same materials showed low activity against P. aeruginosa.

Contaminant-degrading bacteria are super carriers of antibiotic resistance genes in municipal landfills: A metagenomics-based study.

Municipal landfills are hotspot sources of antimicrobial resistance (AMR) and are also important habitats of contaminant-degrading bacteria. However, high diversity of antibiotic resistance genes (ARGs) in landfills hinders assessing AMR risks in the affected environment. More concerned, whether there is co-selection or enrichment of antibiotic-resistant bacteria and contaminant-degrading bacteria in these extremely polluted environments is far less understood. Here, we collected metagenomic datasets of 32 raw leachate and 45 solid waste samples in 22 municipal landfills of China. The antibiotic resistome, antibiotic-resistant bacteria and contaminant-degrading bacteria were explored, and were then compared with other environmental types. Results showed that the antibiotic resistome in landfills contained 1,403 ARG subtypes, with the total abundance over the levels in natural environments and reaching the levels in human feces and sewage. Therein, 49 subtypes were listed as top priority ARGs for future surveillance based on the criteria of enrichment in landfills, mobilizable and present in pathogens. By comparing to those in less contaminated river environments, we elucidated an enrichment of antibiotic-resistant bacteria with contaminant-degrading potentials in landfills. Bacteria in Pseudomonadaceae, Moraxellaceae, Xanthomonadaceae and Enterobacteriaceae deserved the most concerns since 72.2% of ARG hosts were classified to them. Klebsiella pneumoniae, Acinetobacter nosocomialis and Escherichia coli were abundant multidrug-resistant pathogenic species in raw leachate (∼10.2% of total microbiomes), but they rarely carried contaminant-degradation genes. Notably, several bacterial genera belonging to Pseudomonadaceae had the most antibiotic-resistant, pathogenic, and contaminant-degrading potentials than other bacteria. Overall, the findings highlight environmental selection for contaminant-degrading antibiotic-resistant pathogens, and provide significant insights into AMR risks in municipal landfills.

The protective role of commensal gut microbes and their metabolites against bacterial pathogens

ABSTRACT Multidrug-resistant microorganisms have become a major public health concern around the world. The gut microbiome is a gold mine for bioactive compounds that protect the human body from pathogens. We used a multi-omics approach that integrated whole-genome sequencing (WGS) of 74 commensal gut microbiome isolates with metabolome analysis to discover their metabolic interaction with Salmonella and other antibiotic-resistant pathogens. We evaluated differences in the functional potential of these selected isolates based on WGS annotation profiles. Furthermore, the top altered metabolites in co-culture supernatants of selected commensal gut microbiome isolates were identified including a series of dipeptides and examined for their ability to prevent the growth of various antibiotic-resistant bacteria. Our results provide compelling evidence that the gut microbiome produces metabolites, including the compound class of dipeptides that can potentially be applied for anti-infection medication, especially against antibiotic-resistant pathogens. Our established pipeline for the discovery and validation of bioactive metabolites from the gut microbiome as novel candidates for multidrug-resistant infections represents a new avenue for the discovery of antimicrobial lead structures.

Harnessing Bacillus Species for Antibiotic Discovery: A Review

The genus Bacillus is a group of rod-shaped, gram-positive bacteria known for their vast potential of the production of antibiotics. This review examines the potentials of various Bacillus species in the production of antibiotics. There is a global threat of antimicrobial resistance and have spurred the need in scientific research for novel antibiotics and Bacillus species offers a ray of hope in the discovery of novel antibiotics. Notable among the antibiotics produced by Bacillus species include fengycin, gramicidin, iturin, polymyxin, subtilin, difficidin, surfactin, megacin, lichenysin, bacitracin, cerein, pumilicin, circulin, each with greater stability and wide spectrum of activity against gram-positive as well as gram-negative bacteria, fungi. This review highlights the various Bacillus species with antibiotics producing abilities such as B. subtilis, B. polymyxa, B. cereus, B. pumilus, B. megaterium, B. circulans, the antibiotics produced by these Bacillus species, mechanisms of actions of these antibiotics, potential application in medicine, agriculture, biotechnology and industry. Bacillus species with antibiotics producing potentials can be applied in combatting the threat posed by antibiotic-resistant pathogens as well as multi-drug-resistant pathogens.

Cryo-EM structure and complementary drug efflux activity of the Acinetobacter baumannii multidrug efflux pump AdeG.

Multidrug-resistant Acinetobacter baumannii has emerged as one of the most antibiotic-resistant bacterial pathogens associated with nosocomial infection, with its resistance highly depending on multiple multidrug efflux pumps. Here, we report the cryoelectron microscopy (cryo-EM) structure of Acinetobacter drug efflux G (AdeG), the inner membrane component of one of three important resistance-nodulation-cell division (RND) pump family members in A.baumannii, which is involved in drug resistance to chloramphenicol, trimethoprim, ciprofloxacin, and clindamycin. We systematically compare the structures and substrate binding specificities of AdeG, AdeB, and AdeJ multidrug efflux pumps via molecular docking, revealing potential determinants for drug binding. Knockout experiments demonstrate a functional complementarity between AdeABC, AdeFGH, and AdeIJK. Our study provides a structural understanding of A.baumannii multidrug efflux pump AdeG and reveals complementary drug efflux activity between AdeG and other RND efflux pumps, which may promote further rational drug discovery efforts targeting multidrug efflux pumps.

Quorum quenching effects of linoleic and stearic acids on outer membrane vesicle-mediated virulence in Chromobacterium violaceum

Chromobacterium violaceum is a pathogenic bacterium that can infect humans and animals, yet the role of its outer membrane vesicles (OMVs) in mediating pathogenicity remains underexplored. This study evaluated the effects of linoleic acid (LA) and stearic acid (SA) on quorum sensing (QS)-mediated violacein production, biofilm formation, and OMV biogenesis in C. violaceum. Our findings revealed that 2 mM LA and 1 mM SA effectively quench QS, leading to a significant reduction in violacein production, biofilm formation, and OMV biogenesis. Gene expression analysis confirmed the downregulation of QS-related genes, including cviI, cviR, vioA, vioB, and vioC, in fatty acid-treated C. violaceum. Additionally, we assessed the antimicrobial activity of C. violaceum-derived OMVs on Rhizobium sp., a PGPR and observed a marked reduction in bactericidal activity in the treated OMVs. This study suggests that LA and SA have potential as anti-infective agents to mitigate OMV-mediated virulence and combat antibiotic resistance in pathogens.

Aminopeptidase-Responsive NIR Photosensitizer for Precision Targeting and Eradication of Pseudomonas aeruginosa Biofilms.

The emergence of resistance in Pseudomonas aeruginosa represents a significant global health challenge, particularly due to the hurdle of effectively penetrating biofilms with antimicrobials. Moreover, the rise of antibiotic-resistant pathogens has driven the urgent need for developing innovative therapeutic approaches to overcome antibiotic resistance. Antibacterial phototherapy strategies have shown great potential for combating pathogens due to their broad-spectrum antimicrobial activity, spatiotemporal controllability, and relatively low rate of resistance emergence. However, due to the lack of bacterial specificity and penetration, photosensitizers cause considerable damage to mammalian cells and normal tissues and are less effective against bacterial biofilms. Herein, we developed a novel dual-targeting antibacterial strategy to construct a near-infrared photosensitizer, Cy-NEO-Leu. Cy-NEO-Leu showed great bacterial targeting affinity, penetrating and accumulating in biofilms. At the site of infection, it was specifically activated by P. aeruginosa aminopeptidase (PaAP), producing Cy-NEO-NH2, which demonstrated outstanding photothermal (PTT) and photodynamic (PDT) properties, with a photothermal conversion efficiency of up to 70.34%. Both in vitro and in vivo results demonstrated that Cy-NEO-Leu significantly reduced the biofilm biomass and bacterial viability in P. aeruginosa biofilms. Moreover, phototherapy with Cy-NEO-Leu further activated the immune system, enhancing therapeutic efficacy and promoting wound healing. RNA-seq analysis revealed that the antibacterial mechanism of Cy-NEO-Leu-mediated phototherapy involves disruption of the transcriptional and translational processes of P. aeruginosa under laser irradiation. Overall, our results present a promising therapeutic approach against P. aeruginosa biofilms and inspire the development of next-generation antimicrobials.

Antimicrobial Resistance in the Context of Animal Production and Meat Products in Poland-A Critical Review and Future Perspective.

The prevalence of antimicrobial-resistant bacteria in meat and meat products is a significant public health challenge, largely driven by the excessive and inappropriate use of antimicrobials in animal husbandry. In Poland, a key meat producer in Europe, antibiotic-resistant pathogens such as Campylobacter spp., Staphylococcus spp., Enterococcus spp., Listeria monocytogenes, and Enterobacterales have been detected in meat, posing serious risks to consumers. This review examines the use of antimicrobial agents in meat production and the resulting antimicrobial resistance (AMR) in microorganisms isolated from meat products in Poland. The mechanisms of AMR, genetic factors, and prevalence in Poland are presented. It highlights key factors contributing to AMR, such as antibiotic misuse in livestock farming, and discusses the legal regulations governing veterinary drug residues in food. This review emphasizes the importance of monitoring and enforcement to safeguard public health and calls for further research on AMR in the meat industry. Antimicrobial resistance in meat and meat products in Poland is a huge challenge, requiring stricter antibiotic controls in animal husbandry and improved surveillance systems. Additionally, the impact of husbandry practices on the environment and food requires further research. Future efforts should focus on nationwide monitoring, alternative strategies to reduce antibiotic use, and stronger enforcement to combat antimicrobial resistance and protect public health.

Antibacterial, antibiofilm, and gene expression assessment of ajwain (Trachyspermum ammi) essential oil on drug-resistant gastrointestinal pathogens and its combination effect with ampicillin.

Essential oils are natural substances used as therapeutic agents and food preservatives to inhibit harmful microorganisms. This study aimed to assess the synergistic effect of Trachyspermum ammi essential oil and ampicillin on antibiotic-resistant gastrointestinal pathogens, including Escherichia coli, Enterococcus faecalis, Shigella flexneri, and Salmonella serotype Typhimurium. Using gas chromatography-mass spectrometry (GC-MS), the main components of T. ammi essential oil were identified as thymol, gamma terpenes, and cymene. The antibacterial and antibiofilm properties were evaluated by minimum inhibitory concentration (MIC), disk diffusion, and microtiter plate methods, revealing MIC values of 2, 1, 4, and 4mg ml-1 for E. coli, E. faecalis, S. flexneri, and S. Typhimurium, respectively, and inhibition zones between 10 and 14mm. Pathogens were examined for their biofilm-related virulence genes, including aggR, esp, icsA, and fliC, using real-time polymerase chain reaction (RT-PCR) in E. coli, E. faecalis, S. flexneri, and S. Typhimurium, respectively. The methyl thiazole tetrazolium (MTT) assay was used to evaluate the essential oil's effect on the viability of human embryonic kidney 293 (HEK293) cells, which showed cell viability of over 80%. The combination of T. ammi oil and ampicillin demonstrated a synergistic effect, and biofilm formation was inhibited. E. faecalis exhibited the greatest sensitivity, while S. flexneri exhibited the lowest sensitivity.

Effect of colistin-tigecycline combination on colistin-resistant and carbapenem-resistant Klebsiella pneumoniae and Acinetobacter baumannii.

Colistin resistance in carbapenem-resistant Gram-negative pathogens is a serious challenge in clinical settings. Our study showed that low concentration of colistin can be effective when combined with tigecycline. It presents the possibility of an available treatment option for antibiotic-resistant pathogens.

The potential utility of phage therapy in the treatment of periprosthetic infection caused by multidrug-resistant pathogens: a review

Background. One of the main and most costly problems in large joint arthroplasty associated with socially significant losses is periprosthetic infection. Of particular importance is periprosthetic joint infection (PJI) caused by antibiotic-resistant pathogens and followed by the worst treatment results. Currently, when antibiotic-resistant pathogens are spreading rapidly, particularly in health facilities, researchers are constantly looking for alternative treatments for severe cases of PJI. Phage therapy can be one of these methods. The aim of the review was to analyse the application of personalized bacteriophages in the management of periprosthetic infection caused by multidrug-resistant pathogens during arthroplasty of large joints of the lower extremities. Methods. A 20-year literature search (2003-2023) was performed in the eLIBRARY, PubMed, Google Scholar, and Scopus electronic databases. Results. During the search, 1.482 publications meeting the objectives of this review were found. Taking into account the inclusion/non-inclusion criteria adopted by the authors, the review included 12 studies describing 45 cases of the use of bacteriophages in the management of hip and knee PJI caused by multidrug-resistant pathogens. The infection was eradicated without recurrence in 42 cases (89.4%), recurrence was recorded only in 5 cases (10.6%). Conclusions. The analysis of the literature confirms that a combination therapy of personalized phages and antibiotics as adjuvant therapy is safe and effective. Bacteriophage treatment is a promising direction in the fight against resistant pathogens of periprosthetic infection. Its combination with antibiotics seems to be the most promising, as it can significantly increase the effectiveness of therapy for infections caused by multidrug-resistant pathogens due to the synergy of these drug classes.

Highly Sensitive In Vivo Imaging of Bacterial Infections with a Hydrophilicity-Switching, Self-Immobilizing, Near-Infrared Fluorogenic β-Lactamase Probe Enriched within Bacteria.

The emergence of antibiotic resistance, particularly bacterial resistance to β-lactam antibiotics, the most widely prescribed therapeutic agents for infectious diseases, poses a significant threat to public health worldwide. The discovery of effective therapies against antibiotic-resistant pathogens has become an urgent need, necessitating innovative approaches to accelerate the identification and development of novel antibacterial agents. On the other hand, the expression of the β-lactam-hydrolyzing enzyme (β-lactamase), the major cause of bacterial resistance to β-lactam antibiotics, provides a distinctive opportunity to visualize bacterial infection, evaluate the efficacy of existing antibiotics, screen for novel antibacterial agents, and optimize drug dosing regimens in live animals. Herein, a hydrophilicity-switching, self-immobilizing, near-Infrared fluorogenic β-lactamase probe for the highly sensitive imaging of bacterial infection in live mice is reported. This probe, in addition to a significant increase in fluorescence upon selective hydrolysis by β-lactamases as conventional β-lactamase probes, also massively enriches within β-lactamase-expressing bacteria (over 1500-folds compared to the incubation medium), which renders excellent sensitivity in the imaging of bacterial infections in living animals. This agent has proven to enable the assessment of antibiotic therapeutic efficacy and potency of β-lactamase inhibitors in living animals in a non-invasive and much more convenient manner.

Diversity of Endolysin Domain Architectures in Bacteriophages Infecting Bacilli.

The increasing number of antibiotic-resistant bacterial pathogens is a serious problem in medicine. Endolysins are bacteriolytic enzymes of bacteriophages, and a promising group of enzymes with antibacterial properties. Endolysins of bacteriophages infecting Gram-positive bacteria have a modular domain organization. This feature can be used to design enzymes with new or improved properties by modifying or shuffling individual domains. This work is a detailed analysis 1of the diversity of endolysin domains found in bacteriophages infecting bacilli. During the course of the work, a database of endolysins of such bacteriophages was created, and their domain structures were analyzed using the NCBI database, RASTtk, BLASTp, HHpred, and InterPro programs. A phylogenetic analysis of endolysins was performed using MEGA X. In 438 phage genomes, 454 genes of endolysins were found. In the endolysin sequences found, eight different types of catalytic domains and seven types of cell wall binding domains were identified. The analysis showed that many types of endolysin domains have not yet been characterized experimentally. Studies of the properties of such domains will help to reveal the potential of endolysins for the creation of new antibacterial agents.

β-lactam antibiotics induce metabolic perturbations linked to ROS generation leads to bacterial impairment.

Understanding the impact of antibiotics on bacterial metabolism is crucial for elucidating their mechanisms of action and developing more effective therapeutic strategies. β-lactam antibiotics, distinguished by their distinctive β-lactam ring structure, are widely used as antimicrobial agents. This study investigates the global metabolic alterations induced by three β-lactam antibiotics-meropenem (a carbapenem), ampicillin (a penicillin), and ceftazidime (a cephalosporin)-in Escherichia coli. Our comprehensive metabolic profiling revealed significant perturbations in bacterial metabolism, particularly in pathways such as glutathione metabolism, pantothenate and CoA biosynthesis, pyrimidine metabolism, and purine metabolism. Antibiotic treatment markedly increased reactive oxygen species levels, with meropenem reaching nearly 200 ± 7%, ampicillin at 174 ± 11%, and ceftazidime at 152 ± 7%. Additionally, β-lactam antibiotics elevated 8-OHdG levels to 4.73 ± 0.56-fold for meropenem, 2.49 ± 0.19-fold for ampicillin, and 3.19 ± 0.34-fold for ceftazidime; 8-OHG levels increased to 5.57 ± 0.72-fold for meropenem, 3.08 ± 0.31-fold for ampicillin, and 4.45 ± 0.66-fold for ceftazidime, indicating that oxidative stress enhances oxidative damage to bacterial DNA and RNA. Notably, we observed a selective upregulation of specific amino acids associated with cellular repair mechanisms, indicating a metabolic adaptation to counteract oxidative damage. These findings illustrate that β-lactam antibiotics induce a complex metabolic perturbations associated with ROS production, potentially compromising critical cellular components. This study enhances our understanding of the intricate relationship between antibiotic action and bacterial metabolism, providing valuable insights for developing effective strategies against antibiotic-resistant pathogens.

Inducing programmed cell death trough MazEF system to combat Staphylococcus aureus: A non-antibiotic treatment candidate.

The aim of the present study was to evaluate the role of extracellular death factor (EDF) derived from Escherichia coli in the induction of programmed cell death (PCD) in methicillin-resistant and -susceptible Staphylococcus aureus (MRSA and MSSA). The confirmation of bacterial strains as well as the minimum inhibitory concentration (MIC) test were performed according to CLSI, 2022. The extraction and efficacy determination of EDF as well as the CFU assessment were done. The expression of mazE and mazF gens in different conditions was evaluated by Real-time PCR. The likely formation of persister cells from MRSA and MSSA, and the possible synthesis of EDF in old cultures of these pathogens was evaluated, as well. The combination of EDF of two E. coli strains and sub-MIC rifampin reduced the CFUs of MRSA and MSSA strains in mid-logarithmic growth phase while increased the expression of mazF several times more than mazE gene. The expression of these genes in different conditions were unlike. EDF was produced in the old cultures of MRSA and MSSA. The supernatant of E. coli 25922 was more powerful than the clinical strain ones to decrease the CFUs of the MRSA and MSSA. The EDF derived from E. coli in combination with sub-MIC rifampin could induce PCD in MRSA and MSSA through activation of the MazEF system. This phenomenon could be exploited as a non-antibiotic treatment candidate to combat the infections caused by the antibiotic-resistant pathogens. However, more studies should be performed in this regard.

Cell-free supernatant of Lactobacillus gasseri 1A-TV shows a promising activity to eradicate carbapenem-resistant Klebsiella pneumoniae colonization.

The use of beneficial bacteria like Lactobacillus spp. is a potential innovative approach to fight antibiotic-resistant pathogens. Klebsiella pneumoniae is one of the most concerning multi drug-resistant (MDR) pathogens, and its ability to colonize the human gut is considered to be the main reason for recurrent infections in critically ill patients. In this study, Lactobacillus gasseri 1A-TV, already described for its probiotic activity, was characterized at the genomic level. Moreover, its cell-free supernatant (CFS) was tested for antimicrobial activity against extended-spectrum β-lactamase (ESBL)- and carbapenemase (KPC)-producing K. pneumoniae clinical isolates. Whole-genome sequencing showed that the L. gasseri 1A-TV genome was of 2,018,898 bp in size with 34.9% GC content, containing 1,937 putative protein coding sequences, 55 tRNA, and 4 rRNA detected by RAST and classified in 20 functional groups by Cluster of Orthologous Genes (COG). BAGEL4 (BActeriocin GEnome minimal tooL) and the antiSMASH 7.0 pipeline identified two bacteriocin biosynthetic gene clusters (BBGCs), namely, BBGC1 that comprises two class IIc bacteriocins including gassericin A-like bacteriocin, and BBGC2 carrying the class III bacteriocin helveticin J. Strikingly, 1A-TV CFS inhibited the growth of all K. pneumoniae isolates only after 8 h of incubation, showing a bactericidal effect at 24 h and interfering, even at lower concentrations, with the biofilm production of biofilm-producer strains independently of a bactericidal effect. NMR analysis of CFS identified and quantified several metabolites involved in carbohydrate metabolism and amino acid metabolism, and organic acids like ethanol, lactate, acetate, and succinate. Finally, in vitro assays of 1A-TV showed significant co-aggregation effects against carbapenem-resistant K. pneumoniae, namely, strains 1, 2, 3, and 7. Our findings highlight the antimicrobial activity of 1A-TV as a probiotic candidate or its CFS as a natural bioproduct active against MDR K. pneumoniae strains, underlining the importance of novel therapeutic strategies for prevention and control of ESBL- and carbapenemase-producing K. pneumoniae colonization.

Tuning antibacterial efficacy against Pseudomonas aeruginosa by using green AgNPs in chitosan thin films as a plastic alternative

Nanotechnology advancements have facilitated the development of eco-friendly strategies to combat bacterial infections caused by antibiotic-resistant pathogens. This study promotes a green method for the synthesis of silver nanoparticles (AgNPs) utilizing Eucalyptus globulus leaf extracts as an alternative to traditional colloidal AgNPs obtained through chemical synthesis, investigating their antibacterial efficacy against Pseudomonas aeruginosa and their impact on the expression of bacterial virulence factors (pyocyanin, pyoverdine, rhamnolipids). This work demonstrates that: i. while colloidal AgNPs showed ineffective up to 120 μM, green AgNPs had a bactericidal effect already at 20 μM, without impacting bacterial virulence factors at sub-inhibitory concentrations; ii. the polyphenolic shell surrounding green AgNPs could play a crucial role in the antibacterial mechanisms, with a pro-oxidant action confirmed by a greater sensitivity to hydrogen peroxide (H2O2); iii. AgNPs improved the antibacterial properties of chitosan when incorporated into thin films. Consequently, an environmentally friendly nanocomposite film with antibacterial and antibiofilm properties was produced, which holds promise for application in food packaging to mitigate the emergence of microbial contamination in food products.

Plastic Litter: A Hidden Reservoir for Antibiotic-Resistant Pathogens in Coastal Ecosystems

Plastic litter carrying pathogenic bacteria pose serious threats to the health of human and aquatic life. We enumerated the abundance of Escherichia coli and Vibrio species in different types of plastic litter (biomedical, food packages, sanitary, fishing material and miscellaneous) collected from the beaches of Agatti and Kavaratti in the Lakshadweep islands of India using quantitative real-time PCR. Further the antimicrobial resistance profile of E. coli and Vibrio species isolated from these samples were also tested. A very high number (more than three thousand gene copies per gram of plastic litter) of Escherichia coli, and Vibrio spp. were detected in the plastic litter. Vibrio spp. in the plastic litter were identified as V. harveyi, V. neocaledonicus, V. japonicus, V. parahaemolyticus and V. campbelli. More than sixty percentage of E. coli and Vibrio spp. isolated from all samples except the E. coli isolated from fishing gear of Kavaratti had multiple antibiotic resistance index of more than 0.25. Most of the bacteria were resistant towards ampicillin, erythromycin, and moxifloxacin, while least resistance was observed against gatifloxacin, gentamicin, imipenem, meropenem, norfloxacin and trimethoprim. These results indicate that plastic litter may act as a conduit for the attachment and transport of multiple antibiotic resistant pathogens, causing severe setbacks to the global efforts on attaining sustainable development goals 3 (health), 6 (clean water) and 14 (life under water). Therefore, strict implementation of plastic waste management rules, awareness campaigns for cleaning beaches and citizen science initiatives to monitor and remove plastics are proposed for protecting the beaches from the deleterious effects of mismanaged plastics.