Microbial secondary metabolites remain an important source of structurally diverse bioactive compounds with huge therapeutic potential. Yeasts, particularly Candida albicans, possess remarkable metabolic plasticity; thus, it paves the way for identifying antibacterial substances from their extracellular metabolome as it remains underexplored. In the present study, a semi-preparative high-performance liquid chromatography (HPLC) method was developed and optimized for the efficient isolation and purification of hydrocinnamic acid (3-phenylpropionic acid) from the extracellular metabolite extract of C. albicans. The purified compound was characterized using UV-Vis, FTIR, NMR (1H and 13C), GC-MS, and X-ray diffraction analyses. This study represents the first report of hydrocinnamic acid isolated and purified from the extracellular metabolites of C. albicans. The antibacterial potential of the purified hydrocinnamic acid was assessed against multidrug-resistant (MDR) strains of Staphylococcus aureus, Enterobacter cloacae, and Escherichia coli, which exhibited concentration-dependent inhibition against all tested pathogens, with distinct activity against S. aureus and E. cloacae. The minimum inhibitory concentrations were 10 μg/mL for S. aureus, 15 μg/mL against E. cloacae, and 40 μg/mL against E. coli. Collectively, these results underscore the unexplored antibacterial potential of C. albicans-derived extracellular metabolites and position hydrocinnamic acid as a promising natural candidate for combating multidrug-resistant bacteria.

Antimicrobial resistance (AMR) in Enterobacteriaceae poses a critical threat to global health, particularly in regions where antibiotic misuse and limited surveillance prevail. Intensive care units (ICUs) act as focal points for resistance emergence and dissemination. This study aimed to evaluate long-term resistance trends among clinically significant Enterobacteriaceae isolated from two hospitals in North Lebanon (2016-2024). A retrospective analysis was conducted on 14,013 Enterobacteriaceae isolates obtained from Al Monla and Dar Al Shifa hospitals between 2016 and 2024. Identification was achieved using biochemical testing or MALDI-TOF mass spectrometry, and susceptibility testing followed EUCAST standards. No molecular characterization of resistance genes was performed. Data were analyzed using SPSS v23.0, applying Chi-square or Fisher's exact tests (p < 0.05). Among 14,013 isolates, Escherichia coli (E. coli) was predominant (66.4%), followed by Klebsiella pneumoniae (K. pneumoniae) (12.4%), Proteus mirabilis (P. mirabilis) (5.5%), and Enterobacter cloacae (E. cloacae) (3.7%). Overall, β-lactam resistance remained widespread across species. In E. coli, carbapenem non-susceptibility increased from 2.7% to 4.8% (p < 0.001). P. mirabilis and E. cloacae exhibited significant increases in ESBL production (p = 0.006 and p = 0.007, respectively). K. pneumoniae showed fluctuating resistance trends with a mild decline in carbapenem resistance (p = 0.029). ICU isolates demonstrated higher ESBL and MDR rates than non-ICU isolates (p < 0.01), and hospital-acquired infections showed significantly higher resistance across most drug classes compared to community-acquired cases (p < 0.05). Over the 9-year study period, Enterobacteriaceae isolates in North Lebanon demonstrated persistently high β-lactam resistance and increasing carbapenem non-susceptibility, particularly in E. coli. These findings underscore the importance of ongoing AMR surveillance and provide regional data that may support future evidence-based clinical and public health decision-making.

Salad vegetables are essential in our daily diet because they provide affordable nutrients. However, they can serve as vectors for the transfer of antibiotic resistance to consumers. In this study, we demonstrated an inter-farm measurement that provides a unique assessment to better understand the proximate causes of drug resistance in human food chains. Salad vegetable samples (cucumber, carrot and tomato) were collected from three agricultural farms in Kharkhari, Rajshahi, Bangladesh. Isolation of tetracycline-resistant bacteria was carried out by the agar dilution method using nutrient agar supplemented with 32 µg/ml tetracycline at 37ºC for 24 hours. Antimicrobial susceptibility testing was performed by the Kirby-Bauer disc diffusion method on Mueller Hinton agar using commercial antibiotic discs. Resistant isolates were identified by 16S rRNA sequencing. A total of 34 tetracycline-resistant bacteria were isolated randomly from three types of salad vegetables to determine their susceptibility to ampicillin, kanamycin and ciprofloxacin. The result shows three resistance patterns among vegetable isolates in a very close geographical area, suggesting various patterns of drug resistance in environmental bacteria. The bacteria identified are: Enterobacter cloacae, Bacillus cereus, Chryseobacterium spp., Pseudomonas sp., and Enterobacter sp. The presence of antibiotic-resistant bacteria in agricultural products of plant origin may offer valuable information on the transmission of resistant bacteria from farms to consumers via the food chain, raising serious concerns for human health.

In Vitro Activity of Cefiderocol Against Clinically Relevant Gram-Negative Bacilli Collected in China in 2023.

Antimicrobial resistance (AMR) in Neonatal Intensive Care Units (NICUs) is a growing public health concern, particularly due to ESKAPE pathogens, which are major causes of severe healthcare-associated infections. This retrospective cohort study analyzed the incidence, resistance patterns, and risk factors of invasive ESKAPE infections in a Brazilian NICU from 2015 to 2024, using National Healthcare Safety Network surveillance data. Among 2035 admitted neonates, 6.7% developed invasive ESKAPE infections, with incidence increasing from 3.5% in 2015 to 9.2% in 2024. A total of 173 infection episodes were identified, predominantly bloodstream (59.5%) and urinary tract infections (34.7%). Klebsiella pneumoniae (52%) was the most frequent pathogen, followed by Staphylococcus aureus (26.6%) and Enterobacter cloacae (16.8%). As the study included only ESKAPE isolates exhibiting resistance to at least one antimicrobial agent, all isolates had an AMR phenotype; among them, 54.9% classified as multidrug-resistant (MDR) and 3.5% as extensively drug-resistant (XDR). ESBL production was detected in 28.3% of isolates. Prior exposure to broad-spectrum antimicrobials-especially carbapenems (OR = 25.71), glycopeptides (OR = 8.69), and fourth-generation cephalosporins (OR = 8.23)-was significantly associated with infection. Infections prolonged hospital stay and were associated with a 23.5% case-fatality rate, which was more than twice the mortality rate observed among neonates without invasive healthcare-associated infections. Mortality was particularly high in Acinetobacter baumannii infections. These findings underscore the urgent need for strengthened surveillance, infection prevention, and antimicrobial stewardship strategies in NICUs.

The rapid global spread of antimicrobial resistance via β-lactamase (bla) demands targeted strategies that selectively eliminate resistant pathogens without exacerbating resistance. Herein, we report BIN-3I, a photosensitizer (PS) that enables bla-selective activation and-more importantly-covalent retention and accumulation within resistant bacteria, allowing potent photodynamic eradication of bla-expressing pathogens and exerting counter-selection pressure. BIN-3I adopts an enzyme-triggered "one-to-multi" design: Upon bla hydrolysis, it undergoes a hydrophilic-to-hydrophobic and low-to-high permeability transition, facilitating uptake and generating a reactive quinone methide intermediate that covalently binds intracellular proteins, leading to >2,000-fold accumulation within resistant bacteria, thiol depletion and light-triggered reactive oxygen species generation. In vitro, BIN-3I demonstrated bla-specific activation, selective enrichment in bla-expressing MRSA, and potent photodynamic killing (>99.999% reduction), outperforming the corresponding uncaged photosensitizer. Crucially, it selectively eradicates bla-producing MRSA within mixed populations, exerting counter-selection pressure against resistant strains. The PS also activates and accumulates in bla-expressing Gram-negative Enterobacter cloacae, albeit with reduced killing efficiency compared to Gram-positive bacteria. In vivo, it exhibited prolonged retention at infection sites and targeted imaging capability in murine myositis and abscess models. Notably, BIN-3I-mediated photodynamic therapy effectively cleared MRSA infections across multiple models-including thigh infection, wound, and biofilm-associated infections-achieving >5-log reduction in bacterial load and surpassing vancomycin in efficacy. This work presents a targeted antimicrobial platform that exploits bacterial resistance mechanisms to achieve species-specific eradication, offering a promising strategy to combat multidrug-resistant infections and alleviate the selection pressure that drives the enrichment of resistant strains.

Antibiotic-resistant bloodstream infections are a major public health concern, particularly in children under five years of age. The objective of this study was to determine the prevalence of bloodstream infections and assess the antimicrobial resistance profiles and their associated factors among febrile children under five years of age in Bahir Dar, northwest Ethiopia. A cross-sectional study was conducted from November to December 2025 among 281 children under five years of age who attended the pediatric clinic of Felege Hiwot Comprehensive Specialized Hospital in Bahir Dar. Sociodemographic and clinical data were collected using a structured questionnaire through face-to-face interviews with the parents or guardians. Approximately 3 mL of venous blood was aseptically collected and cultured using standard microbiological techniques. Bacterial identification was performed based on colony characteristics and biochemical tests. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method. Data were entered into EpiData version 3.1 and analyzed using SPSS version 26. Logistic regression analysis was used to identify factors associated with bloodstream infections, and statistical significance was set at p < 0.05. The prevalence of bloodstream infection was 19.2% (54/281; 95% CI: 14.0%-24.4%). The most common isolate was Klebsiella pneumoniae (31%), followed by Enterobacter cloacae (11%), Escherichia coli (9%), and Acinetobacter baumannii (9) %. Gram-negative bacteria accounted for 80% of the isolates. Antimicrobial susceptibility testing revealed high resistance rates to commonly used antibiotics, particularly ampicillin (87.0%, 40/46), trimethoprim-sulfamethoxazole (86.3%, 44/51), and cefepime 42/54 (77.8). Resistance was also high to ciprofloxacin (68.5%, 37/54), tobramycin (68.6%, 35/51), amoxicillin-clavulanic acid (66.0%, 31/47), gentamicin (63.3%, 31/49), tetracycline (61.1%, 33/54), and chloramphenicol (60.5%, 26/43). In contrast, lower resistance rates were observed for cefotaxime (37.0%, 20/54) and meropenem (20.4%, 11/54). Significant associated factors of blood stream infections included age < 1 year (AOR = 2.11; 95% CI: 1.46-2.86), fever duration > 7 days (AOR = 2.74; 95% CI: 1.22-6.15), partially immunized children (AOR = 3.21; 95% CI: 1.08-9.51; p = 0.036), and non-immunized children (AOR = 9.87; 95% CI: 3.61-26.9; p < 0.001). The prevalence of bloodstream infections among febrile children under five years of age was high. Younger age, fever duration > 7 days, and partially immunization and non-immunization were significant predictors of bloodstream infection. Strengthening antimicrobial stewardship, improving immunization coverage, and enhancing early diagnosis are essential to reduce the burden of bloodstream infections and combat antimicrobial resistance.

Azo dyes are widely used synthetic colorants characterized by one or more azo (-N = N-) bonds and are major contributors to environmental pollution due to their persistence and toxicity in industrial effluents. In the present study, indigenous bacterial strains were isolated from textile wastewater and evaluated for their potential to degrade reactive violet (RV) dye. A total of fifteen isolates were obtained through enrichment, among which four strains namely, Enterobacter cloacae, Stenotrophomonas geniculata, Pseudomonas inefficax, and Stenotrophomonas maltophilia demonstrated significant dye decolourization efficiency (>75%) at 200 ppm and were selected for further studies. Among these, P. inefficax exhibited the highest decolorization efficiency (up to 86%) under optimized conditions (pH 7, 25 °C, 2% inoculum, 400 ppm dye concentration, and 150 rpm) within 48 h. Fourier-transform infrared (FTIR) analysis indicated the cleavage of azo bonds in the dye molecules after incubation. Chemotaxis, bioaccumulation, and biofilm formation assays revealed adaptive traits that enhanced bacterial interactions with dye molecules and improved degradation efficiency. To the best of our knowledge, no previous study has evaluated the degradation of RV dye by P. inefficax while simultaneously investigating chemotaxis, biofilm formation, and bioaccumulation traits associated with dye removal. Overall, this study demonstrates the metabolic versatility and significant biodegradation potential of P. inefficax, which can be used as a promising candidate for the treatment of dye-contaminated textile effluents.

Genomic analysis of an extensively drug-resistant Enterobacter cloacae NH77 from Thailand carrying an ∼82-kb chromosomal heavy-metal and antibiotic resistance island.

We report the draft genome sequence of Enterobacter cloacae TG-C5.2 isolated from shrimp aquaculture water in Vietnam. The genome harbors multiple antimicrobial resistance genes, including blaCTX-M-15.

BackgroundThe effectiveness of antibiotic therapy for urogenital tract infections (UTIs) is increasingly compromised by the global dissemination of extended‐spectrum β‐lactamase‐producing Enterobacterales (ESβL‐En). This study aimed to assess the involvement of broad‐spectrum β‐lactamase‐producing Enterobacterales (BSβL‐En) in UTIs and to establish their antibiotic susceptibility profile.MethodsA cross‐sectional study was conducted from October to December 2019 at the Army Training Hospital Omar Bongo Ondimba. All patients suspected of having a UTI were included. BSβL‐En isolates were recovered using MacConkey agar supplemented with cefotaxime. Phenotypic screening for β‐lactamase (ESβL, AmpC, and carbapenemases) using specific phenotypic confirmatory disc diffusion test. Antimicrobial susceptibility was determined using the disk diffusion method and minimum inhibitory concentration (MIC) assays. The presence of resistance genes (blaCTX-M, blaTEM, blaCARBA, blaIMP, mcr-1 and mcr-2) was investigated by polymerase chain reaction.ResultsA total of 295 patients were enrolled, among whom 18.3% (54/295) had UTIs. Of the isolated uropathogens 74.1% were BSβL‐En. The predominant species were Enterobacter cloacae (8/54, 14.8%), Escherichia coli (6/54, 11.1%) and Kluyvera spp (5/54, 9.3%). Among 40 BSβL‐En isolates, 87.5% (n = 35/40) were confirmed by molecular analysis. Notably 48.5% (17/35) harbored at least two bla genes, simultaneously. Fourteen isolates carried the blaCARBA and blaIMP genes, which in combination with the blaCTX-M and blaTEM. About 40.0% of the isolates (14/35) carried the blaCTX-M, 8.6% had the blaTEM (3/35), and 2.9% had the blaIMP (n = 1/35). All urinary isolates were multidrug‐resistant (MDR), exhibiting high levels of β‐lactamase production, with resistance rates of 81.6% ± 0.16% for first‐line antibiotics (e.g., β‐lactams, fluoroquinolones) and 61.7% ± 0.04% for last‐resort antibiotics (e.g., imipenen, ertapenem, and colistin). The prevalence of carbapenem resistance genes was 44.1%, whereas no colistin resistance genes (mcr‐1 and mcr‐2) were detected.ConclusionThis study highlights a high prevalence of multidrug‐resistant uropathogens producing BSβL, with significant resistance to both first‐line and last‐resort antibiotics.

Background:Antimicrobial resistance (AMR) in Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae is a global threat, yet data from Belize and the wider Caribbean are scarce. Understanding local resistance patterns is critical for effective treatment and stewardship.Objectives:To determine antibiotic susceptibility patterns, quantify multidrug resistance (MDR) in outpatient and inpatient settings, identify predictors of empirical treatment failure for urinary tract infections (UTIs), and analyze temporal and geographic trends.Design:Nationwide retrospective analysis of clinical isolates from Belize’s public health system (2018–2023).Methods:We analyzed 4591 clinical isolates. MDR was defined as non-susceptibility to ⩾3 drug categories. Multivariable logistic regression identified predictors of the risk of empirical treatment failure for UTIs. Joinpoint regression analyzed temporal trends. Geospatial analysis mapped hotspots of incidence and resistance.Results:E. coli was predominant (68%). Among 3005 urinary isolates, 1247 (41.5%) were resistant to both first-line oral antibiotics (ciprofloxacin and trimethoprim-sulfamethoxazole). Extended-spectrum β-lactamase (ESBL) production (adjusted odds ratio (aOR) 8.91, 95% CI 7.40–10.73), infection with K. pneumoniae (aOR 2.85, 95% CI 2.32–3.51) or E. cloacae (aOR 4.12, 95% CI 2.78–6.11), and older age were key predictors. Ciprofloxacin resistance in E. coli increased significantly post-2020 (Annual Percent Change 9.8%, p = 0.02). MDR was prevalent in outpatient settings (42.1% for E. coli, 61.2% for K. pneumoniae). Geospatial analysis revealed the Belize District as the incidence hotspot, while the Toledo District was the ESBL prevalence hotspot (29.5%).Conclusion:Belize faces a notable AMR crisis characterized by a high community burden of MDR Enterobacterales, a critical empirical treatment gap for UTIs, and accelerating resistance trends.

Diverse structures of mcr-10-bearing plasmids and high colistin resistance in Enterobacter cloacae complex clinical isolates from South Korea.

In agricultural applications, plant growth-promoting bacteria (PGPB) have demonstrated their efficacy as both biofertilizers and biocontrol agents against several phytopathogens. This study reports, for the first time, the isolation of an endophytic Enterobacter cloacae from the medical plant Portulaca oleracea L., which produces iturin-like lipopeptides effective against B. Sorokiniana that causes severe diseases that affects several important crops. The present study investigates the ability of E. cloacae as a PGPB and a producer of bioactive metabolites that participate in the biological control of Bipolaris sorokiniana. Twelve endophytic bacterial isolates obtained from P. oleracea, grown in El-Sharkia Governorate, Egypt, were examined for their antagonistic activity against B. sorokiniana through the disk diffusion method. Also, the most active isolates (2, 5, 7 and 8) were investigated using a dual culture technique against B. sorokiniana in vitro, and the percentages of inhibition were 43.48, 40.55, 36.60, and 35.57%, respectively. Therefore, the most active endophytic bacterial isolate (2) was molecularly identified as E. cloacae strain RO2 with accession number PV739238. In vitro assessment of PGP traits of E. cloacae strain RO2 proved that it can produce protease, amylase, cellulase, in addition to nitrogen fixation, siderophore production, phosphate and calcium carbonate solubilization and ammonia production. E. cloacae caused irregular mycelial growth and morphological deformation in B. sorokiniana hypha. HPLC analysis of E. cloacae’s lipopeptide extract revealed the occurrence of lipopeptide like Iturin that had inhibitory effects on B. sorokiniana. In addition, the growth-promoting potential of E. cloacae was evaluated on wheat (Triticum aestivum L.) seeds bio-primed with it, and the results showed enhanced shoot height, radicle length, fresh and dry weight of seedling as compared to the control. Consequently, E. cloacae strain could be used as biofertilizers and biocontrol agent in agriculture to reduce the risk of chemical pesticides.

ObjectivesNacubactam is a novel diazabicyclooctane carbapenemase inhibitor that, in combination with cefepime or aztreonam, has been submitted for regulatory approval in Japan for treating serious Gram-negative infections. This study aimed to evaluate the in vitro activity of nacubactam combined with cefepime or aztreonam against a nationwide collection of clinical carbapenem-resistant Enterobacterales (CRE) isolates in Japan.MethodsMinimum inhibitory concentration (MIC) values of 376 CRE isolates from 6 medical institutions in Japan were determined according to the Clinical and Laboratory Standards Institute (CLSI) methods. Results were interpreted using clinical breakpoints of the CLSI document M100 (2025) and the European Committee on Antimicrobial Susceptibility Testing clinical breakpoints, version 15.0 (2025).ResultsThe predominant CRE isolates were Klebsiella pneumoniae (n = 98), Klebsiella aerogenes (n = 62) and Escherichia coli (n = 43), with the Enterobacter cloacae complex accounting for 117 isolates. Carbapenemase-producing Enterobacterales (CPE) accounted for 48.9% (n = 184), with the predominant carbapenemases being IMP-1 (n = 70), IMP-6 (n = 43) and NDM (n = 17). Cefepime–nacubactam and aztreonam–nacubactam exhibited potent antibacterial activity against CRE isolates with MIC50/90 of 1/4 and 0.5/2 mg/L, respectively. Cefepime–nacubactam and aztreonam–nacubactam demonstrated potent activity against CPE, with MIC50/90 of 2/4 and 0.5/2 mg/L, respectively, which were lower than those for ceftazidime–avibactam (64/>64 mg/L) and imipenem–relebactam (2/16 mg/L). Both combinations exhibited potent antibacterial activity against non-carbapenemase-producing carbapenem-resistant Enterobacterales (non-CP-CRE), with MIC50/90 of 0.25/4 and 0.5/4 mg/L, respectively.ConclusionsCefepime–nacubactam and aztreonam–nacubactam have excellent antibacterial activities against CPE and non-CP-CRE, supporting their potential as therapeutic options for CRE infections.

Bacterial infections have become the second leading cause of death worldwide, and antimicrobial resistance (AMR) is a major factor contributing to treatment failure. Rapid and accurate bacterial identification (ID) and antibiotic susceptibility testing (AST) are critical for improving patient outcomes and controlling the spread of resistant strains. In this study, we developed a novel, rapid, and accurate diagnostic method for bacterial infections based on metabolic volatile organic compounds (VOCs). First, we performed untargeted profiling of metabolic VOCs from 9 clinically prevalent pathogens (Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas aeruginosa, Acinetobacter baumannii, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, and Enterococcus faecium) using proton transfer reaction mass spectrometry (PTR-MS). Notably, all 10 species (including the culture medium control) could be successfully distinguished using only four VOCs: isovaleraldehyde, methanethiol, acetaldehyde, and dimethyl disulfide. The entire ID workflow only takes 3 h, achieving 98.0% accuracy. And then, we established a phenotypic AST method leveraging bacterial activity-related characteristic VOCs (ARC-VOCs). Testing the susceptibility of these 9 pathogens to different antibiotics achieved 99.1% accuracy (455 bacterial-antibiotic combination), with a turnaround time of 3-4 h. In summary, this metabolic VOC-based method enables bacterial infection diagnosis (ID + AST) within 6-7 h. It cuts the diagnostic time of the widely used clinical platform VITEK 2 by more than 50%, while maintaining high accuracy. This approach holds promise for guiding clinicians to develop timely personalized treatment regimens and curbing the transmission of drug-resistant bacterial strains.

AI-optimized sanguinarine derivatives inhibiting sortase A for combating AmpC β-lactamase resistance in Enterobacter cloacae: An integrated computational approach.

Multidrug-resistant (MDR) Enterobacter cloacae has emerged as a serious public health threat, particularly in hospital settings, due to its ability to cause severe infections and rapidly acquire antibiotic resistance. The limited effectiveness of current antibiotics highlights the urgent need for alternative preventive strategies such as vaccination. In this study an immunoinformatics-based approach was employed to design a multi epitope vaccine (MEV) targeting the conserved and surface exposed outer membrane protein A (OmpA) of MDR E. cloacae. The entire proteome was screened to identify suitable vaccine targets based on antigenicity, allergenicity and homology analysis. Highly immunogenic B-cell, MHC class I and MHC class II epitopes were predicted and selected for vaccine construction. The selected epitopes were linked using suitable linkers, while the 50S ribosomal L7/L12 protein was incorporated as an adjuvant. The designed MEV construct was evaluated through structural modelling, molecular docking with TLR4 receptor, immune simulation, population coverage analysis, codon optimization and in silico cloning. The vaccine candidate demonstrated strong antigenicity, non-allergenicity, structural stability, and favourable physicochemical properties. Population coverage analysis indicated 99.92% global coverage. Molecular docking revealed stable and energetically favourable interactions with TLR4, while immune simulation predicted strong humoral and cellular immune responses, including memory cell formation and a Th1-biased cytokine profile. Codon optimization indicated favorable expression potential in an E. coli. Overall, the proposed MEV predicted a promising, safe, stable and broadly protective vaccine candidate against MDR Enterobacter cloacae.

Fabrication, characterisation and antimicrobial application of the ZnO@fGO nanocomposite

Extraction and characterization of chitin, chitosan, and nanochitosan from black soldier fly (Hermetia illucens) pupal exoskeletons: insights into structural, thermal, microstructural, color, and antimicrobial properties.