Cell elongation and cytokinesis are mediated by two large multi-protein complexes in rod-shaped bacteria: the elongasome and divisome. These membrane-associated complexes ultimately recruit peptidoglycan cell wall remodeling and synthesis machineries, thus sculpting and extending growing cells or driving septation of two daughter cells. The elongasome and divisome have been well studied in model bacterial species, including Bacillus subtilis, Caulobacter crescentus, and Escherichia coli. Here, we present an analysis of these complexes in an obligate intracellular bacterium with a highly reduced genome. Orientia tsutsugamushi, a cytoplasm-dwelling Gram-negative alphaproteobacterium, only retains a subset of proteins normally found in the elongasome and divisome. It also lacks all copies of the major peptidoglycan polymerase, Class A Penicillin Binding Protein, and has been shown to build an intermediate peptidoglycan cell wall-like structure that is low in abundance and insufficient to consistently confer a rod shape to the bacterium. We have carried out a first analysis of the elongasome and divisome in Orientia tsutsugamushi, quantifying the expression and subcellular localization of five key proteins through early stages of the intracellular infection cycle. We show how these are affected by antibiotic treatment and present a model for minimal elongation and division in an obligate intracellular bacterium.

Direct-Fed Microbials in Diets of Market Pigs Did Not Impact Salmonella Levels, Shiga Toxin-Producing Escherichia coli, Growth Performance, and Carcass Characteristics.

The construction of minimal-genome microbes offers an ideal platform for understanding fundamental biological processes and synthetic biology, yet the research is hindered by incomplete lists of essential genes in microbes and by multiple rounds of genome trimming with a trial-and-error nature. To address this, we introduce CREAT (CRISPR-based genome trimming with a multi-homology-arm template)-a streamlined approach that integrates CRISPR-targeted genome cleavage and homology arm walking to classify essential from non-essential genomic subregions, thus providing the basis for predicting essential genes in a given organism. These essential genes were then assembled into synthetic gene cassettes for one-step replacement of the targeted non-deletable genomic regions for further genome trimming. Eight consecutive rounds of CREAT genome trimming achieved a 20.8% reduction in genome size in Saccharolobus islandicus. Furthermore, Cas9-based CREAT genome trimming was developed for Bacillus subtilis and Escherichia coli, with efficiency greatly enhanced by the λ-Red recombinase in the latter. Together, this iterative application of CREAT provides a scalable and generally applicable strategy for rapidly constructing minimal genomes across diverse microorganisms.

Class-Incremental Learning for Foodborne Pathogen Prediction Based on Clinical Surveillance Data.

AimsRecombinant human bone morphogenetic protein-2 (rhBMP-2) is used clinically for bone regeneration. The Food and Drug Administration-approved clinical product (Infuse) is Chinese hamster ovary (CHO)-derived, but Escherichia coli (EC)-derived rhBMP-2 may offer manufacturing advantages. This study compared CHO- and EC-derived rhBMP-2 potency in vitro and in vivo.MethodsFour EC-derived rhBMP-2 preparations were screened using alkaline phosphatase (ALP) assays in C2C12 and W20-17 cells. The most potent (EC-MAX) was compared to CHO-rhBMP-2. A total of 15 male athymic rats received bilateral absorbable collagen sponge (ACS) implants loaded with 17 µg rhBMP-2 (CHO or EC-MAX) in muscle pouches for four weeks. Bone formation was assessed by radiography, micro-CT, and histology.ResultsEC-MAX showed 2.2-fold higher osteogenic activity in W20-17 cells and 1.3-fold in C2C12 cells versus CHO-rhBMP-2. In vivo, EC-MAX produced ~ ten-fold greater bone volume (88.5 mm³ (SD 12.8) vs 9.2 mm³ (SD 5.4)) and tissue volume (203.3 mm³ (SD 47.9) vs 15.0 mm³ (SD 8.8)). Histology confirmed more mature bone architecture in EC-MAX implants with adipocyte-filled marrow spaces.ConclusionPotency cannot be generalized across products. Inter-supplier potency variation exists among EC-derived rhBMP-2 products. EC-MAX demonstrated superior osteoinductive potential both in vitro and in vivo versus CHO. However, substantial variability highlights the need for product-specific characterization.Cite this article: Bone Joint Res 2026;15(6):686–695.

Effects of combined Lactococcus lactis and Escherichia coli Nissle 1917 cell-free supernatants on metabolic activity and inflammation-associated signaling in HT-29 cells.

Dysbiosis of the gut microbiota in patients with chronic kidney disease (CKD) causes an increase in proteolytic bacteria and a decrease in saccharolytic bacteria, leading to an increase in the production of uremic toxins such as indoxyl sulphateeeee, which will induce inflammation through the gut-kidney axis. The research objective is to analyse the effect of administering Trigona honey on the number of Bifidobacterium Sp., Lactobacillus Sp., Clostridium Sp., Escherichia coli, Indoxyl Sulfat levels, and IL-6 before and after intervention in patients with chronic kidney disease. The method used was a double-blind randomised controlled trial for 60 days on CKD patients aged 18-60 years, stage 3b-5 non-dialysis, who had not consumed prebiotics, probiotics, synbiotics, antioxidants, and antibiotics in the past month. Dietary education was provided for 14 days and participants were randomised into 2 groups, namely the treatment group (n=16), receiving trigona honey, and the control group (n=18), receiving a placebo. Both groups received the intervention for 60 days. Blood biochemical tests (WBC, Hb, and platelets), urea, creatinine, GFR, Indoxyl Sulfat levels, IL-6, urine albumin creatinine ratio, and stool examinations to assess the number of Bifidobacterium Sp., Lactobacillus Sp., Clostridium Sp., and and and and and Escherichia coli ore and after the intervention were conducted. From the research results, 40 patients were randomised into 20 patients in the treatment group and 20 patients in the control group. At the end of the study, there were 16 patients in the treatment group and 18 patients in the control group. The gender variable in both groups had more males (61.76%) compared to females (38.24%). In the age variable, the average age of patients in the treatment group was 44.75±9.40 years, while in the control group it was 45.94±6.94 years. There were no significant differences in the average food intake of energy, protein, fibre, carbohydrates, and fat composition between the two groups, both before and after the intervention. There was a significant decrease in the number of Bifidobacterium sp. in the treatment group compared to the control group (p=0.025 vs p=0.269). There were no significant changes in the number of Lactobacillus sp., Clostridium sp., E. colii, IS levels, and IL-6 in both groups. The conclusion of this study is that It can be said that in CKD patients, trigona honey greatly lowers the amount of Bifidobacterium sp. but does not affect the amounts of Lactobacillus sp., Clostridium sp., E. coli, IS levels, and IL-6.

The emergence of antibiotic-resistant microbes requires the development of superior antimicrobial materials. This study focuses on the chemical modification of xanthan gum (XG) by free-radical graft copolymerization with butyl acrylate (BuA) initiated by potassium persulfate (KPS), to significantly improve its structural, thermal, and functional characteristics. The relationship between reaction parameters and grafting performance was comprehensively analyzed, focusing on grafting yield (G), homopolymer formation (H%), and grafting efficiency (GE). Optimal grafting conditions (0.6 M BuA, 8 × 10–2 M KPS, 60 °C, and 120 min) produced a graft copolymer with up to a 500% grafting yield. Zinc oxide nanoparticles (ZnO NPs) were manufactured in situ and integrated into the grafted copolymer matrix, resulting in the formation of the xanthan gum grafted by butyl acrylate with zinc oxide (XG-g-BuA/ZnO) nanocomposite. The structural, thermal, morphological, and crystalline properties were investigated utilizing FTIR, TGA, SEM/EDX/mapping, TEM, and XRD to validate successful grafting and incorporation of ZnO nanoparticles into the copolymer matrix. Thermal studies showed that the grafted XG displayed improved thermal stability compared to its unmodified counterpart, with thermal resistance increasing with increasing BuA content. The antibacterial efficacy was evaluated against Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) and Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae), demonstrating substantial inhibitory effects, with the nanocomposite attaining up to 87.6%, 42.9%, 85.7% and 72.6% reduction in bacterial growth against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Klebsiella pneumoniae, respectively. These outcomes highlight the potential of the fabricated biopolymer nanocomposite as a sustainable and efficient platform for antimicrobial applications in both packaging and biomedical fields.Supplementary InformationThe online version contains supplementary material available at 10.1186/s13065-026-01856-4.

Pseudouridine (Ψ) modifications are the most abundant RNA modifications; however, their distribution and functional significance in bacteria remain largely unexplored compared to eukaryotic systems. In this study, we present the first transcriptome-wide and quantitative mapping of Ψ modifications across five diverse bacterial species (Bacillus cereus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Pseudomonas syringae) at single-base resolution, utilizing the optimized baBID-seq method for bacterial RNA. Our analysis revealed growth phase-dependent dynamics of pseudouridylation in bacterial tRNA and mRNA, particularly in genes enriched in core metabolic pathways. Comparative analysis demonstrated evolutionarily conserved features of Ψ modifications, such as dominant motif contexts, Ψ clustering within operons, etc. Functional analysis indicated Ψ modifications affect bacterial mRNA stability, translation, and interactions with specific RNA-binding proteins in response to changing cellular demands during growth phase transitions. The integrated computational analysis on local RNA architecture was conducted to elucidate the structure-dependent Ψ modifications in bacterial RNA. Furthermore, we developed an integrated deep learning framework, combining LSTM-transformer-GNN-based neural networks (pseU_NN) to capture both RNA sequence and local structure features for effective prediction of Ψ-modified sites. Overall, our study provides valuable insights into the landscapes of bacterial RNA Ψ modifications and establishes a foundation for future mechanistic investigations on bacterial Ψ functions.

Surveillance of Enterobacteriaceae, Escherichia coli, and STEC in Food from the Piedmont Region (Italy): Results of Official Control Activities (2022-2024).

Enhancing agrochemical efficiency through their integration as building block in MOFs

In-Plant Validation of Postharvest Water Sanitation Practices to Prevent Cross-Contamination in Michigan Commercial Apple Packinghouses.

ABSTRACTFood safety in basic schools across developing countries poses significant public health challenges. Soya kebabs, popular protein‐rich snacks sold in Ghanaian schools, have received limited microbiological safety assessment despite widespread consumption. This study investigated the microbial quality of soya kebabs sold in basic schools (covering primary and junior high education, grades 1–9) within Sunyani Municipality, Ghana, examining relationships between vendor hygiene practices and contamination levels. A cross‐sectional study was conducted across 25 basic schools from March to June 2025. Soya kebab samples (n = 50) were collected from 25 vendors at two time points (early week: Monday–Tuesday; late week: Thursday–Friday) and analyzed for total aerobic counts, coliforms, Enterobacteriaceae, Escherichia coli, Staphylococcus aureus, and fungi using standard microbiological methods. Vendor hygiene assessments were performed using structured observation checklists for all 25 vendors. Bacterial contamination levels varied substantially throughout the school week. Total aerobic counts ranged from 4.18 ± 0.13 to 4.71 ± 0.20 log10 CFU/g, while S. aureus contamination ranged from 2.28 ± 0.13 to 4.12 ± 0.10 log10 CFU/g (p < 0.001). E. coli levels ranged between 1.85 ± 0.16 and 2.43 ± 0.19 log10 CFU/g. Public schools showed significantly higher contamination than private schools across multiple parameters. Vendor assessments revealed critical deficiencies: only 16% demonstrated adequate hand washing, 28% had hand washing facilities, and 12% maintained proper temperature control. A strong negative correlation (r = −0.780, p < 0.001) existed between hygiene scores and contamination levels. The study reveals substantial quality variation in soya kebabs throughout the school week and widespread hygiene deficiencies posing significant health risks to students. Targeted interventions addressing vendor training and infrastructure provision could substantially reduce these food safety risks.

Human papillomavirus (HPV) infection, cervical intraepithelial neoplasia (CIN), pelvic organ prolapse (POP), cervical polyps, abnormal uterine bleeding, and vaginitis are common gynecological disorders. This study characterized the vaginal microbiome and its cytokine-mediated immune interactions in Chinese women with gynecological disorders. Vaginal swabs from 310 patients diagnosed with gynecological disorders and 112 healthy controls were analyzed via 16S rRNA gene sequencing and cytometric bead array (CBA) to evaluate proinflammatory cytokines. Findings on correlation were validated through in vitro co-culture experiments. HPV infection and CIN were associated with dysbiotic microbial profiles and elevated levels of IL-1α, IL-1β, IL-6, IL-8, MCP-1, and MIG. Stratification by microbial composition demonstrated that HPV/CIN patients with non-Lactobacillus-dominant microbiomes harbored the highest IL-1α and IL-1β concentrations. Validated through computational modeling and in vitro analyses, Lactobacillus crispatus and Lactobacillus iners were identified as being strongly inversely correlated with IL-1α and IL-1β expression. Conversely, non-Lactobacillus taxa, including Bifidobacterium breve, Prevotella bivia, Gardnerella vaginalis, Sneathia amnii, Sneathia sanguinegens, Prevotella amnii, Escherichia coli, and Chlamydia trachomatis, exhibited positive correlations with proinflammatory cytokines. These findings highlight a connection between Lactobacillus species and reduced inflammation, while non-Lactobacillus dominance is linked to increased cytokine-driven responses. This study provides valuable insights into microbiome-immune crosstalk in gynecological diseases.

BackgroundAntimicrobial stewardship programs (ASPs) are coordinated efforts designed to promote the judicious use of antimicrobials, decrease microbial resistance, reduce the spread of multidrug-resistant infections, and improve patient outcomes. This study aimed to evaluate the impact of an ASP on antimicrobial consumption and antimicrobial resistance in a tertiary cardiac centre in Egypt.MethodsA quasi-experimental study was conducted in the adult critical care unit of Aswan Heart Centre, Magdi Yacoub Foundation, Aswan, Egypt to assess the changes in antimicrobial consumption and resistance patterns before and after ASP implementation. All patients admitted to the adult critical care area from July 2023 to 30 June 2024 were included in the study.ResultsA total of 1000 patients were included in the pre-intervention period and 982 in the post-intervention period. Following ASP implementation, overall antimicrobial consumption decreased by 23% (from 14,414 to 11,012 DDDs per 1,000 patient days), although this was not statistically significant (p = 0.097). Colistin use declined significantly (817 to 53 DDDs per 1,000 patient days; p = 0.006). Infections caused by Klebsiella pneumoniae decreased significantly (42 to 19 isolates; p = 0.004). Among Enterobacteriaceae, Escherichia coli showed improved susceptibility to gentamicin (36% to 86%; p = 0.01) and meropenem (64% to 100%; p = 0.02). Reductions in fungal infections and improvements in antifungal susceptibility were observed, but were not statistically significant. The number of documented fungal infections decreased from 12 pre-intervention (9 Candida spp. 3 Cryptococcus spp.) to 6 post-intervention (4 Candida spp. 2 Cryptococcus spp.) (p = 0.236). Infections caused by Staphylococcus spp. also decreased slightly with improved susceptibility patterns (p = 0.683).ConclusionThe implementation of an ASP in the critical care unit was associated with reductions in antibiotic consumption and trends toward decreased antimicrobial resistance, highlighting its potential value in addressing the challenges of antimicrobial misuse and resistance in resource-limited settings.Clinical trial numberNot applicable.Supplementary informationThe online version contains supplementary material available at 10.1186/s12879-026-13358-8.

Heterologous Expression, Refolding, and Functional Characterization of Halocin C8 from Halophilic Archaea in Escherichia coli.

Bacteria-responsive DNAzymes have been widely used in pathogen detection. We report a "one stone, two birds" strategy that uses one selected DNA molecule to develop two DNAzymes for targeting two bacterial species. Through in vitro selection, we isolated an RNA-cleaving fluorogenic DNAzyme, brRFD2, that can be activated by Klebsiella pneumoniae (KP) and Escherichia coli (EC) under different reaction conditions. Target assessing experiments indicated that RNases from KP and EC function as the activating components for brRFD2, but they differed in properties: the KP RNase is acid-tolerant, while the EC RNase is heat-tolerant. By exploiting these differences, brRFD2 exhibited a strong response to KP at pH 3.6, with a kobs ∼6600-fold higher than that for EC, while showing high activity to EC at pH 4.5 using pre-heated bacteria, with a kobs ∼520-fold higher than that for KP. brRFD2 also demonstrated outstanding performance in the diagnosis of hospital-acquired pneumonia (HAP) caused by KP and EC, achieving an 88.9% sensitivity and 96.7% specificity for KP and 100% sensitivity and specificity for EC across 48 clinical samples. This work demonstrates that a single DNAzyme sequence can function as two different bacteria-responsive DNAzymes by simply regulating the reaction conditions.

The choice of substrates and their composition play a crucial role in supporting the growth of microorganisms and enhancing overall microbial fuel cells (MFCs) performance. Traditionally used substrates, such as glucose and acetate, are now being replaced by innovative alternatives like biomass materials and food waste. Consequently, this research focuses on the use of cow dung as an organic substrate in MFCs, examining its role in microbial metabolism, bioenergy production, and the bioremediation of toxic metals. The MFC processes were conducted over 40 days, during which a peak voltage output of 485 mV was recorded on day 26. The MFC cycle generates maximum metal bioremediation efficiencies of 89.16 ± 0.04% for Al3+, 89.17 ± 0.46% for Cu2+, 92.86 ± 0.11% for Pb2+, 91.37 ± 0.4% for Zn2+, and 92.01 ± 0.16% for Hg2+, respectively indicating its potential in ameliorating toxic metal levels from wastewater. Two-way ANOVA revealed that bioremediation efficiency (%) was significantly influenced by metal ion type (p < 0.001) and operational duration (p < 0.001), with a significant Metal × Operational Days interaction (p < 0.001), indicating metal-specific temporal responses. Post-hoc Tukey HSD analysis confirmed significant differences among most metal-day combinations, demonstrating progressive, time-dependent enhancement of bioremediation efficiency. The substrate was applied to the anodic region constantly throughout the experiment. Cyclic voltammetry (CV) revealed distinct redox peaks, confirming active bioelectrochemical processes, while electrochemical impedance spectroscopy (EIS) indicated reduced internal resistance and enhanced electron transfer efficiency. SEM-EDX revealed the extent of biofilms and toxicity level of the operation. Furthermore, microbial analysis was investigated on the anode electrode after MFC operation. The probable bacterial species identified were Bacillus subtilis, Providencia rettgeri, Escherichia coli, Serratia marcescens, Staphylococcus aureus, Morgarella morganii and Providencia alcalifaciens. This study has shown that cow dung is a potential substrate for generation of electric current and bioremediation of toxic metals in MFC.

BackgroundEnterotoxigenic Escherichia coli (ETEC) remains a major cause of diarrheal disease in low- and middle-income countries (LMICs). To curb ETEC related diarrhoea, several candidate vaccines are in development, with ETVAX® being the most advanced. Although immunogenicity studies have primarily focused on measuring antibody titres, assessing antibody avidity offers additional valuable insight into antibody quality and immune maturity. This study assessed anti-CS3 and anti-LTB IgG avidity in Zambian children to better understand vaccine-induced antibody responses in an endemic setting.MethodsChildren aged 6–23 months (n = 60) received three quarter-doses of ETVAX® with dmLT adjuvant on days 1, 14, and 90. Plasma samples collected at baseline prior to the first vaccination (Day 1; V1), seven days after the second dose (Day 22; V5), and seven days after the third dose (Day 97; V7) were analysed by limiting antigen dilution ELISA to calculate avidity indices (AI). Naïve classification was performed using titre-based thresholds (20th percentile of baseline titres) and avidity-defined naivety (AI < 0.5). Receiver operating characteristic (ROC) analysis was used to evaluate the discriminatory performance of avidity indices against titre-defined naïve status.ResultsBaseline avidity was detectable for both CS3 and LTB, consistent with prior natural exposure. Mean CS3 IgG avidity decreased from 0.7 at baseline to 0.6 after the third dose (p < 0.001), while LTB IgG avidity showed transient decreases but no net gain. Naïve classification at baseline revealed that 9/60 children had titres but low avidity (functional naivety), and 6/60 had waned titres but high avidity. Only one child was naïve by both criteria for CS3, and none for LTB. ROC analysis demonstrated moderate discrimination for CS3 (AUC = 0.65; optimal cut-off AI = 0.36) but poor discrimination for LTB (AUC = 0.30).ConclusionIn this endemic population, ETVAX® induced strong antibody titres but minimal changes in avidity over time, with notable inter-individual variability. ROC analysis highlighted context-specific limitations in using avidity to discriminate immune maturity. Together, these findings suggest that while antibody avidity may provide complementary information on antibody quality, its interpretation should be cautious and considered alongside antibody titres in endemic settings.

We investigated the interaction of liquid crystal droplets, formed by a phase separating system of elongated virus particles and polymer with rod-like impurities. The virus particles were Pf4, which when mixed with sodium alginate as polymer, phase separated into an isotropic phase in coexistence with a nematic phase of the virus particles. The nematic phase appeared in the form of spindle-shaped liquid crystal droplets, which are called tactoids. We studied the interaction of this mixture with micrometer sized rods, which were either bacteria (Bacillus subtilis or Escherichia coli) or inert colloidal particles (made of SU8 photoresist polymer or silica). Confocal laser scanning microscopy was used to examine the mixtures and four different dominant configurations were found, classified as no attachment, partial attachment of the rod to the tactoid, a "sandwich" state, where two tactoids interact with the same rod, and lastly encapsulation of the rods by the tactoids. We categorized the results in terms of surface properties and rod geometry (size and aspect ratio). We further investigated the system through numerical calculation in a simplified two-dimensional model. Finally, we addressed the role of confining the experimental system from three dimensions to a quasi two-dimensional setup, a relevant scenario in bacteria biofilms.