Fusobacterium spp. are Gram-negative, obligate anaerobic bacteria associated with a broad clinical spectrum, including head and neck infections; soft tissue infections; gastrointestinal and genitourinary infections; and bacteremia without an identified source. Clinical manifestations vary by species and site of colonization. F. necrophorum is linked to head and neck infections that may culminate in Lemierre syndrome, a life-threatening septic thrombophlebitis of the internal jugular vein. Although Fusobacterium bacteremia is uncommon, it is associated with substantial mortality. F. necrophorum bacteremia may occur as a complication of head and neck infection in younger, healthy individuals, whereas F. nucleatum bacteremia is reported predominantly in older patients with malignancy, secondary to an abdominal source or without an identified origin. Fusobacterium isolates are usually susceptible to penicillins, cephalosporins, aminopenicillins with β-lactamase inhibitors, carbapenems, and metronidazole, while resistance to clindamycin and moxifloxacin has been increasingly reported. Because susceptibility testing is not routinely performed and susceptibility data remain limited, severe infections are commonly treated with a β-lactam/β-lactamase inhibitor, a carbapenem, or a β-lactam in combination with metronidazole. This review provides an overview of the microbiology, clinical spectrum, and treatment of Fusobacterium spp.
Representatives of the phylum Methanobacteriota occur in various anoxic environments, but only members of the genera Methanosphaera and Methanobrevibacter exclusively colonize the digestive tract of animals. Recent phylogenomic analyses revealed that the genus Methanobrevibacter, which harbors the majority of the intestinal species, is severely underclassified and represents a family-level taxon, "Methanobrevibacteraceae", that evolved entirely in the digestive tract of animals. Comparative genome analysis of 158 species of Methanobacteriota, including uncultured representatives in the Genome Taxonomy Database (GTDB), demonstrated that the intestinal lineages are clearly separated from the remaining members of the phylum. They differ from the non-intestinal lineages in genome size, GC content, coding density, an increased number of pseudogenes and adhesin-like proteins, and show numerous adaptations to the copiotrophic gut environment. A decreased biosynthetic potential led to a dependence on other community members and limits the dispersal of intestinal species into other habitats, which is reflected in coevolutionary patterns with their major host groups among arthropods, ungulates, and primates. Certain lineages even engaged in symbiotic associations with intestinal protists, presumably benefiting from the H2 produced by the hydrogenosomes of their anaerobic hosts. Our results reveal that the transition of free-living Methanobacteriota to a host-associated lifestyle involves the same genomic changes that were previously recognized in gut bacteria and bacterial endosymbionts of protists, reflecting resemblances between the two prokaryotic domains that are caused by evolutionary convergence in similar environments.
The transformation products (TPs) of pharmaceuticals and pesticides are ubiquitous in aquatic environments, necessitating the development of advanced treatment technologies for effective removal. This study systematically evaluated the degradation efficiency of six typical TPs of carbamazepine and atrazine by UV222-based advanced oxidation processes (AOPs) combined with different oxidants (periodate, hydrogen peroxide, peroxymonosulfate, persulfate, and chlorine). The results indicated that the oxidants were efficiently activated under 222 nm irradiation to generate diverse radical species, thereby accelerating TPs degradation. The dominant reactive species varied significantly across different treatment systems. Notably, the high photon energy of UV222 facilitated substantial ozone formation (6.5 × 10-9-1.61 × 10-6 M•cm2•mW-1), and potentially promoted electron transfer through the formation of excited states of oxidants and TPs. For CBZ-EP, the degradation pathways under UV222/oxidant treatments involved amide hydrolysis, epoxide hydrolysis/oxidation, decarboxylation, dehydrogenation, and aromatization. Economic analysis further confirmed the feasibility of UV222-based technologies. Moreover, the presence of dissolved organic matter (DOM) inhibited TPs removal in the UV222-AOPs, while concurrently altering the optical properties of DOM. DOM underwent dealkylation, oxygenation, and decarboxylation pathways, resulting in the formation of more saturated and highly oxidized molecules, along with the potential generation of disinfection by-products. This study provides insights into the mechanisms of TPs removal by UV222-AOPs and the molecular-level transformation of coexisting DOM, offering guidance for the optimization of advanced wastewater treatment processes.
Traditional Chinese medicine (TCM) products frequently face adulteration risks, demanding rapid, field-deployable authentication methods. We developed a novel duplex Proofman ladder-shape melting temperature isothermal amplification (Proofman-duplex-LMTIA) assay that simultaneously discriminates Schisandra chinensis (authentic ingredient) from Schisandra sphenanthera (common adulterant) by targeting species-specific single nucleotide polymorphisms (SNPs) in the ITS1 region. Incorporating proofreading enzyme-mediated probe cleavage to enhance specificity, the assay uses dual fluorescence reporting (FAM/JOE) in a single closed-tube reaction at a constant temperature of 60 °C for 20 min. The duplex format achieved a detection limit of 10 fg/μL plasmid DNA for both targets, with no cross-reactivity against related plant species. It reliably detected adulteration levels as low as 0.1% (S. sphenanthera in S. chinensis) and 1% (vice versa) in binary mixtures. Validation using 13 commercial TCM products demonstrated 100% agreement with labeling. This isothermal method offers a practical, equipment-minimal molecular tool that complements chromatographic fingerprinting for on-site quality assurance in TCM supply chains.
Toxoplasma gondii is an important opportunistic pathogen in people living with HIV (PLWH), and the risk of reactivation increases markedly when CD4 counts fall below 100/mm³ . We aimed to determine T. gondii seroprevalence in PLWH, assess its associations with demographic and clinical variables, and emphasize preventive strategies for seronegative individuals. This retrospective cohort study analyzed data from 1419 people living with HIV who were diagnosed by Western blot and followed at a tertiary care center between January 2006 and January 2024.Demographic characteristics, HIV RNA levels, CD4 counts, education level, probable route of HIV transmission, and T. gondii IgG/IgM results at the time of diagnosis were reviewed. Associations between categorical variables were evaluated using the chi-square test, and independent predictors were assessed using logistic regression analysis. Of the 1419 patients, 88.2% were male; the mean age was 45.7 years and the mean CD4 count was 302.6/mm³ . T. gondii IgG seropositivity was 41.1%. Seropositivity was significantly higher among patients aged ≥ 50 years and those with CD4 < 100/mm³ according to the chi-square test (p < 0.001). Heterosexual individuals had higher seropositivity, whereas men who have sex with men (MSM) were more often seronegative. Higher education level was significantly associated with seronegativity. This study, conducted in one of Türkiye's long-standing HIV care centers, provides comprehensive data on T. gondii seroprevalence among PLWH. The high seronegativity observed among younger and more educated individuals underscores the need for targeted education on hygiene and food safety. Regular screening and appropriate prophylactic strategies in seronegative patients at risk of advanced immunosuppression are critical to prevent life-threatening toxoplasmosis reactivation.
River networks are critical pathways for antibiotic discharge and the spread of antimicrobial resistance (AMR). This study investigates the spatial and temporal distribution of AMR in sediments of the Qiantang river estuary, a typical transitional zone between terrestrial and marine ecosystems. Seven sampling sites were strategically established along the estuary. Major antibiotic resistance gene (ARG) classes, including multidrug, aminoglycoside, tetracycline, bacitracin, and macrolide-lincosamide-streptogramin (MLS) resistance genes were analyzed. Nine high-risk Rank I ARG subtypes were identified, with horizontal gene transfer (HGT) emerging as a key mechanism for ARG dissemination. A strong positive correlation between ARG abundance and mobile genetic elements (MGEs) (R2 = 0.856, Pearson's r = 0.925), highlighting the role of transposons and integrons in facilitating ARG propagation within sedimentary microbial communities. Sediment bacterial hosts, particularly Proteobacteria (e.g., Phenylobacterium, Thioalkalivibrio, Cupriavidus) and Chloroflexi (e.g., Anaerolinea), were strongly associated with Risk Rank I genes such as aac(3)-I and aac(3)-IV, encoding aminoglycoside acetyltransferases (AACs). The co-occurrence patterns of ARGs with hosts and human pathogens in sediments suggest that ARG subtypes such as golS, ompR, and ksgA are most closely related to their hosts. Notably, antibiotic concentrations in water and sediments correlated positively with both MGEs and ARG diversity, emphasizing the synergistic effects of anthropogenic pollutants. This study provides new insights into the occurrence and transmission of ARGs in estuarine environments and offer an omics-based assessment strategy for the public health risks associated with AMR.
Oleanolic acid (OA) is a natural anti-inflammatory triterpenoid. This study investigates its therapeutic potential against rheumatoid arthritis (RA), a chronic autoimmune condition characterized by inflammatory activity, and explores the novel mechanism of gut microbiota modulation in a rat model. The anti-arthritic effect of OA was assessed in collagen-induced arthritis (CIA) in rats following oral or intravenous administration. Arthritis severity, joint pathology, and mesenteric Th17/Treg cell frequencies were evaluated. The role of gut microbiota was investigated using 16S rRNA sequencing and antibiotic ablation. Oral, but not intravenous, OA administration significantly alleviated arthritis and joint damage in CIA rats, indicating a gastrointestinal-dependent mechanism. Therapeutically, oral OA modulated gut microbiota by reducing Prevotella abundance and restored immune balance by decreasing Th17 and increasing Treg cell frequencies. This protective effect was abolished by co-treatment with antibiotics, confirming gut microbiota-dependency. Taken together, our findings show that the improvement in CIA after oral administration of OA is mainly due to changes in the structure and function of the gut microbiota. This study, therefore, suggests that OA is a promising therapeutic candidate for RA, based on its ability to restore the intestinal microenvironment.
Portal hypertension (PH) is one of the major complications of liver cirrhosis, traditionally assessed using invasive methods such as the hepatic venous pressure gradient (HVPG). Soluble endoglin (sENG), a marker of endothelial dysfunction and fibrosis, has been proposed as a non-invasive biomarker of various liver diseases. This study aimed to evaluate serum sENG concentrations in cirrhotic patients with PH and investigate its relationship with PH severity, alcohol consumption, and smoking. Serum concentrations of sENG were measured in clinically well-examined patients with liver cirrhosis (n = 60, age range 24-82 years) with PH classified as mild, moderate, or severe according to the HVPG values measured invasively using the classical wedge technique. sENG concentrations were also compared to healthy controls (n = 54). Liver enzyme activities, alcohol consumption history, and smoking habits were also recorded to assess their association with sENG. sENG concentrations were significantly higher in patients with PH compared to healthy controls (6.31; 5.14-7.30 vs. 3.70; 3.24-4.20 ng/mL, p < 0.001) but did not correlate with the severity of HVPG-diagnosed portal hypertension. A moderately significant correlation was observed between sENG concentrations and GGT activities (p < 0.001). Alcohol consumption, but not smoking, was associated with higher serum sENG concentrations (p < 0.01). Based on our results, sENG appears to be a non-invasive marker of endothelial dysfunction/fibrosis in cirrhotic patients with PH, particularly in alcohol-related liver disease. Although it does not reflect PH severity and thus cannot be used as a diagnostic tool, it has the potential for early disease detection and risk prediction as a screening component in non-invasive approaches in clinical hepatology.
Gyroviruses are widespread small DNA viruses infecting poultry and other hosts, yet their diversity and coinfection dynamics in backyard systems remained poorly understood. Tissue samples from 100 clinically healthy backyard chickens across five provinces in Türkiye, between 2020 and 2021, were screened using species-specific PCR assays targeting six gyroviruses. The Gyroviral DNA was detected in 90.0% of samples (90/100), with CAV identified in 71.0% (71/100), AGyV2 in 59.0% (59/100), GyV3 in 8.0% (8/100), GyV4 in 22.0% (22/100), GyVTu789 in 19.0% (19/100), and GyV7-SF in 18.0% (18/100) of samples. Co-infections were frequent, most notably between CAV and AGyV2, with a co occurrence rate of 25.6%. Phylogenetic analysis of partial VP1 sequences revealed that circulating strains were largely consistent with global lineages, while also displaying detectable genetic variability, particularly within GyV4. Notably, GyVTu789 and GyV7-SF were identified for the first time in chickens in Türkiye, extending their known geographic range. The high rate of co-detection highlights the concurrent circulation of multiple gyroviruses in backyard poultry populations. These findings provide a comprehensive overview of gyrovirus diversity and circulation dynamics, pinpointing the need for continued surveillance to better understand their epidemiology and potential impact on poultry health.
Bone remodelling is essential for maintaining skeletal integrity by preserving the balance between bone formation and resorption, with excessive osteoclast activity contributing to osteoporosis. Osteocytes act as central regulators of osteoclastogenesis through mechanically sensitive paracrine signals, yet the influence of osteoblasts and their mesenchymal precursors remains less defined. Extracellular vesicles (EVs) have recently emerged as mediators of bone cell communication, although their role in osteoclast regulation are still underexplored. This study demonstrates that mesenchymal-derived bone cells inhibit osteoclastogenesis through an EV-dependent mechanism shaped by their differentiation stage and mechanical environment. Mechanically stimulated osteocyte-derived EVs showed the strongest anti-catabolic response. Notably, we identify miR-150-5p as a mechano-responsive miRNA enriched within osteocyte EVs, capable of inducing a dose-dependent reduction in osteoclastogenesis. Transcriptomic analyses reveal that EV treatment and miR-150-5p delivery induce substantial transcriptional changes in osteoclast precursors, including downregulation of shared target genes linked to bone remodelling. Overall, we highlight mechanically activated osteocytes as key regulators of osteoclastogenesis through an EV-mediated mechanism, in which miR-150-5p represents a promising candidate contributor within the broader EV cargo landscape, highlighting their potential for future cell-free therapeutic strategies.
The urobiome of renal transplant recipients is poorly defined. The purpose of this study was to investigate whether there are characteristic changes in the urobiome between pre- to post-transplant states, at varying degrees of post-transplant allograft function, and between those with acute T-cell mediated rejection (TCMR) versus a non-rejector cohort. 41 patients who consented to have urine stored in our transplant biobank were included in this study: 1) Rejectors (n = 10 pts: 6 borderline, 1 Banff IA, 3 Banff IIA TCMR, mean age: 47.4 ± 12.4 yrs); 2) Women (n = 16 pts; mean age: 49.3 ± 17.3 yrs); 3) Men (n = 15 pts, mean age: 47.5 ± 17.2 yrs). Urine was collected via mid-stream clean-catch technique prior to transplant (n = 21), at the time of TCMR (n = 9; within 1 month of transplant), 1-month (n = 15), and 3-months post-transplant (n = 38). Samples were processed and stored at -80 [Formula: see text] until 16S rRNA sequencing. Alpha diversity, beta diversity, and differential abundance analysis was performed. The urobiome was altered post-transplant, with rejectors gaining Corynebacterium and Pseudomonas at time of rejection, and non-rejectors gaining Lactobacillus among other taxa. Within individuals, post-transplant urobiome composition was ~ 75% dissimilar from pre-transplant (p < 0.001). Urobiome composition differed by sex (p = 0.002), but not by age. Differential abundance analysis based on 3-month post-transplant eGFR revealed consistent loss of Lactobacillus with decreased renal function. Our results suggest that renal transplantation has a strong impact on individual urobiome composition, but not diversity, and microbial imbalance may be associated with acute rejection and post-transplant renal function. Our findings indicate a need for further research into the urobiome during renal transplantation to elucidate its potential as a biomarker of and/or contributor to post-transplant allograft health.
Burkholderia pseudomallei, an environmental Gram-negative saprophyte is the causative agent of melioidosis, a potentially fatal yet treatable clinical entity. We report an unusual colony morphology of the organism to minimize misidentification and enable accurate diagnosis.
The objective of this study is to conduct a comparative morphological analysis of the kidneys in the context of various infection pathways in models of acute obstructive pyelonephritis. The study investigated the structural condition of rabbit kidneys (n=40) in relation to the pathogenesis of acute pyelonephritis. The animals were randomly assigned to four groups: two experimental and two control groups. Experimental Groups I and II underwent urethral ligation to induce obstructive pyelonephritis. In Group I, a bacterial strain was introduced into the gastrointestinal tract to examine intestinal translocation, while in Group II, it was introduced into the urinary bladder to study ascending infection. Control Groups III and IV received a similar introduction of the infectious agent without urethral obstruction. On the third day, morphological examination was conducted using optical microscopy and a computerized microscope capable of digital microphotography. In Group I, inflammatory changes were detected in 60% of cases, while in Group II, they were found in 70% of cases. No morphological changes were observed in the control groups. The nature of morphological changes in the kidneys in the model of acute obstructive pyelonephritis did not show statistically significant differences when comparing between the experimental groups. This indicates the involvement of enterorenal bacterial translocation in the development of inflammatory processes in the kidneys in the presence of obstruction.
Blood-borne viral infections pose considerable health challenges to patients undergoing hemodialysis. This multicenter study evaluated the prevalence and demographic distribution of hepatitis B, hepatitis C, and HIV infections among hemodialysis patients. This retrospective cross-sectional study included 940 hemodialysis patients from fifteen facilities in Khartoum State, Sudan from July 2025 to November 2025. The outcomes of the serological tests for hepatitis B surface antigen (HBsAg), anti-hepatitis C virus (anti-HCV) antibodies, and anti-HIV antibodies, conducted via standard enzyme-linked immunosorbent assay (ELISA) test, were acquired from the Public Health Laboratory (PHL), Khartoum State. Data was analyzed with R software (version 4.4.2). The overall prevalence rates were 7.1% for HBsAg, 12.2% for anti-HCV, and 0.6% for HIV. The prevalence of hepatitis C virus (HCV) exhibited significant variability across dialysis centers (p<0.001), with higher rates observed in certain facilities. No notable correlations between infection status and age or gender were detected. Co-infections were rare. Blood-borne viral infections persist as a significant issue in Sudanese hemodialysis facilities, emphasizing the imperative for consistent infection control measures such as routine blood screening for transfusion. Targeted interventions at high-prevalence centers are essential for decreasing transmission and enhancing patient outcomes. Not applicable.
Chronic urticaria (CU) is a complex inflammatory skin disorder characterized by recurrent wheals and pruritus lasting longer than six weeks, often resulting in significant impairment in quality of life, sleep disturbance, and psychological distress. Although the majority of cases are classified as chronic spontaneous urticaria without identifiable triggers, a growing body of literature suggests that persistent infections may contribute to disease pathogenesis through immune-mediated mechanisms. Helicobacter pylori infection has been increasingly investigated because of its ability to induce chronic systemic inflammation, autoantibody production, and mast cell activation. We present a case of a 34-year-old male with a two-year history of recurrent urticaria that progressed to near-daily episodes over a three-month period. The patient also reported chronic dyspeptic symptoms including heartburn and epigastric discomfort. Laboratory investigations revealed normal complete blood count parameters, normal liver function tests, normal total IgE levels, and mildly elevated C-reactive protein. Stool antigen testing confirmed H. pylori infection and endoscopy demonstrated chronic gastritis. Following completion of eradication therapy, the patient remained symptom-free for two consecutive months, with subsequent stool antigen testing confirming successful eradication. This report highlights the potential pathogenetic role of H. pylori infection in selected patients with chronic urticaria and emphasizes the importance of targeted evaluation based on clinical presentation.
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The effects of anaerobic digestates on soil microbial communities have received increasing attention due to their potential impacts on soil health and antibiotic resistance. To date, no integrated analysis of rhizosphere bacterial community structure, antibiotic resistance genes (ARGs), and mobile genetic elements has been conducted in digestate-treated perennial ryegrass (Lolium perenne L.). We analyzed rhizosphere bacterial communities of this pasture using metabarcoding to study the effects of a manure-derived digestate on community structure and predicted functions. We also explored the association between digestate-enriched taxa and explanatory variables, including the abundance of two ARGs, class 1 integrons, and IncP-1ε plasmids. The greenhouse study included an unfertilized control and three fertilization treatments: digestate, inorganic fertilizer, and combined fertilizer (digestate + inorganic fertilizer). The results indicated a significant effect of the fertilizer type on bacterial communities and a stimulation of predicted functions related to genetic information processing by digestate and its combination. Digestate application resulted in the greatest differentiation in bacterial community structure relative to the unfertilized control and shifted communities toward amplicon sequence variants (ASVs) positively associated with class 1 integrons. Differential abundance analysis identified three ASVs and three genera (Arenimonas, Algoriphagus and Novosphingobium) that were significantly enriched under digestate treatment, relative to both urea and the unfertilized control. Our results demonstrate that anaerobic digestate application alters bacterial community structure and highlight the need for further studies to elucidate the potential adaptive role of class 1 integrons in rhizosphere microbiomes following digestate fertilization, including their contribution to antibiotic resistance.
Anaerobic gut bacteria have highly evolved functions that promote transmission between human hosts. These include resilient spores produced by many Bacillota (formerly Firmicutes) bacteria, which are tolerant to extended ambient oxygen exposure. Recent culturing and genomic studies have revealed the taxonomic diversity of gut spore-formers and their prevalence in human populations. However, due to a reliance on spore-forming model organisms that are not representative of commensal gut bacteria, we still have a limited understanding of gut microbiome-specific sporulation processes and their underlying genetics. In this opinion article, we outline a rational path for development of new model organisms derived from commensal spore-forming gut bacteria in order to obtain fundamental insights into sporulation and to provide a discovery platform for novel probiotic or microbiome-based therapeutic development.
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The high Arctic, particularly Spitsbergen, faces the combined challenges of climate change and other anthropogenic pressures - including waste and contaminant release from human activity - that influence microbial populations and the spread of antimicrobial resistance (AMR). This study presents a snapshot analysis of metagenomes from various environments across Spitsbergen, including untreated and treated wastewater outflows, fjords, and glacial ice cores, to explore the abundance of stress-response genes, including antibiotic resistance genes (ARGs), biocide resistance genes (BRGs), metal resistance genes (MRGs), and virulence genes (VGs), alongside the compositions of the associated bacterial populations. We reveal varying levels of stress-response genes and VGs in environments exposed to differing levels of human influence. ARGs and MRGs dominate in raw sewage, while VGs are more prevalent in fjord waters receiving both raw sewage and effluent, indicating that specific environmental conditions favor different resistance and virulence traits. We detect high abundance of ARGs and VGs downstream of both untreated and treated wastewater. Our analyses indicate the presence of bacterial populations with resistance and virulence traits - including Enterobacteriaceae, Enterococcaceae, Bacillaceae, and Staphylococcaceae - in downstream ecosystems. While we do not directly assess effects on human health or ecosystem function, these observations point to potential ecological impacts in Arctic environments and highlight the importance of continued monitoring to understand and manage the possible effects of human activities and climate change.