Chlamydia psittaci pneumonia is often underdiagnosed due to nonspecific presentation and limited diagnostic tools, with scarce data on the comparative utility of metagenomic next-generation sequencing (mNGS) versus targeted next-generation sequencing (tNGS) in this setting. This study aimed to characterize the clinical features of C psittaci pneumonia and provide comparative data on mNGS and tNGS, with emphasis on diagnostic accessibility, while also quantifying underreported features such as hyponatremia. A retrospective analysis was conducted on 10 patients with C Psittaci pneumonia admitted to the Department of Respiratory and Critical Care Medicine at the Second People's Hospital of Hunan Province between January 2022 and February 2025. Clinical data, laboratory tests, imaging findings, treatment regimens, and outcomes were collected and analyzed descriptively. Among the 10 patients (6 male, 4 female; median age 70 years), 9 had underlying comorbidities and 2 had severe pneumonia. A history of avian exposure was reported in 40% of cases. All patients presented with fever (mean temperature: 39.5 ± 0.7°C), accompanied by nonspecific symptoms including cough (n = 4), chills (n = 5), anorexia (n = 5), and myalgia (n = 3). Laboratory findings revealed normal white blood cell counts in 8 cases, while lymphocytopenia, elevated inflammatory markers, abnormal liver enzymes, and hyponatremia were observed in 9 patients. Imaging predominantly showed unilateral lung consolidation (n = 5) or ground-glass opacities (n = 3), with pleural involvement in 9 cases. mNGS and tNGS were performed on bronchoalveolar lavage fluid in 5 cases each; tNGS provides a more cost-saving and rapid alternative. Post-diagnosis, treatment with moxifloxacin, doxycycline, or omadacycline for 10 to 21 days yielded generally favorable outcomes. C psittaci pneumonia is characterized by high fever and systemic symptoms, with imaging typically showing unilateral consolidation, ground-glass opacities, or mass-like lesions with pleural involvement, accompanied by liver injury, elevated inflammatory markers, and hyponatremia. Both tNGS and mNGS are effective diagnostic tools, and tNGS provides a more cost-saving and rapid alternative. Early treatment with fluoroquinolones or tetracyclines confers significant therapeutic benefits.

The objective of this study was to examine the depletion kinetics of enrofloxacin and doxycycline in quail eggs and develop suitable withdrawal times. Four hundred healthy laying Chinese White-feathered quails were randomly assigned to five groups (n = 80 each): a control group and two dose groups (high and low) for each drug. The doses were set at the maximum dose recommended for chickens and at half of that dose, according to the Chinese Veterinary Pharmacopeia. Both drugs were administered continuously for 5 days via drinking water. Eggs were collected at different time points during the administration period and after withdrawal, until residues fell below the detection limit. During the administration period, residues in quail eggs increased progressively, with the highest accumulation found in albumen. After drug withdrawal, the elimination patterns were similar across the dose levels. Residues in albumen declined more rapidly than in yolk. For both drugs, residue concentrations declined rapidly during the initial period after withdrawal and more slowly thereafter. Using the whole-egg residue data and the Chinese national standard (GB 31650.1-2022) maximum residue limits (MRLs) of 10 µg/kg, the withdrawal times were calculated as follows: for enrofloxacin, 10.75 days (0.15 g/L) and 9.03 days (0.075 g/L); for doxycycline, 25.56 days (0.3 g/L) and 21.30 days (0.15 g/L). These results provide valuable data for residue monitoring and risk management, and offer a scientific reference for food safety assurance of quail eggs.

Antimicrobial resistance (AMR) in Salmonella enterica from aquatic animals represents an emerging food safety and public health concern, yet the global distribution patterns and trends remain insufficiently understood. Here, we performed a global genomic survey of Salmonella from aquatic animals using 2911 genomes collected from 67 countries across six continents between 1974 and 2025. These genomes represented 194 serovars and 364 sequence types. Notably, the dataset exhibited unbalanced geographical sampling, with Asian isolates accounting for 69.98% of all strains, which may limit the generalizability of our global trends. Globally, resistance to WHO-recommended clinically important antimicrobials showed substantial variation among antibiotic classes, with resistance rates for azithromycin (2.10%), cefepime (2.89%), cefotaxime (2.71%), ceftazidime (2.71%), and ceftriaxone (0.62%). Geographical differences were observed, with isolates from Asia exhibiting higher resistance levels, particularly for ciprofloxacin (13.50%) and doxycycline (18.02%). AMR temporal trends differed across geographical regions. A total of 111 acquired ARGs and 10 resistance-associated chromosomal mutations were identified, accompanied by changes in the distribution of resistance determinants. Collectively, this study provides a global genomic perspective on priority AMR in Salmonella from aquatic animals and highlights the importance of integrating aquaculture, environmental, and public health surveillance to monitor and mitigate AMR risks within the One Health framework.

Ehrlichia chaffeensis is an obligate intracellular bacterium that proliferates within monocytes or macrophages and causes human monocytic ehrlichiosis (HME), an emerging life-threatening zoonosis. Doxycycline is the choice of treatment for HME, yet it has prominent side effects. Host cells lack the lysine biosynthetic pathway; thus, the enzymes in this pathway are essential for bacterial growth and recognized as potential targets for the development of novel antibiotics. Here, we demonstrated that inhibitors targeting DapE, which is a key enzyme in the lysine biosynthetic pathway, especially disulfiram, effectively inhibit E. chaffeensis infection and intracellular growth. Through complementation experiments and peptide nucleic acid-mediated dapE knockdown, we showed that DapE in E. chaffeensis is functional and essential for bacterial intracellular growth. Using purified recombinant protein, we found that DapE induces IL-8 expression in host cells. Finally, we identified that NtrX, the response regulator of the two-component system NtrY/NtrX, regulates dapE expression using an electrophoretic mobility shift assay and a reporter assay. Our findings deepen the understanding of E. chaffeensis pathogenesis as well as illustrate that DapE in E. chaffeensis is a potential therapeutic target for the development of novel HME treatments.

Epidermal growth factor receptor variant III (EGFRvIII) is a constitutively active EGFR deletion variant found in a subset of glioblastomas (GB). Its association with stem-like tumor populations is well documented, but the outcome of EGFRvIII expression may vary with the developmental and molecular state of the recipient cell. Here, we used human induced pluripotent stem cell (iPSC)-derived, glial fibrillary acidic protein (GFAP)-negative-induced neural stem cells (iNSCs) to explore how this receptor variant affects a defined neural stem/progenitor background. EGFRvIII was introduced either constitutively by lentiviral transduction or through a doxycycline-responsive Tet-On system. Both experimental approaches were associated with remodeling of the neural stem/progenitor marker profile, with reduced SRY-box transcription factor 2 (SOX2) expression in the inducible model and significant reductions in both SOX2 and nestin in the constitutive model. Constitutive expression was also associated with increased senescence-associated β-galactosidase (SA-β-Gal) activity, while doxycycline induction produced a distinct change in cell-number dynamics at low doxycycline concentration. Thus, in this experimental setting, EGFRvIII was linked to substantial remodeling of the SOX2+/nestin+ marker profile. These observations do not define a differentiation pathway, prove cellular senescence or malignant transformation, or identify the cell of origin of EGFRvIII-positive GB. Rather, they support the view that the cellular response to EGFRvIII should be considered in relation to the specific neural-glioblastoma precursor context.

Introduction: Severe periodontitis, corresponding to Stage III and Stage IV disease according to the 2018 World Workshop classification, represents a substantial global health burden. Systemic antibiotics have been investigated as adjuncts to mechanical periodontal therapy; however, their effectiveness and appropriate use in severe periodontitis remain uncertain. This umbrella review aimed to synthesize the available evidence on adjunctive systemic antibiotics in Stage III and Stage IV periodontitis and assess the methodological quality of the available systematic reviews. Methods: This umbrella review was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A systematic search of MEDLINE, Embase, and other relevant databases was conducted from June 2018 to June 2026. Systematic reviews evaluating adjunctive systemic antibiotics in patients with Stage III or Stage IV periodontitis, classified according to the 2018 World Workshop, were eligible. Methodological quality was assessed using AMSTAR-2 and overlap among primary studies was evaluated using the Corrected Covered Area (CCA). Results: Two systematic reviews met the inclusion criteria: a network meta-analysis by Lin et al. and a systematic review by Jones and Hoyle focusing on azithromycin. The CCA was approximately 52.9%, indicating substantial overlap between the included reviews. Lin et al. reported moderate-certainty evidence that adjunctive amoxicillin plus metronidazole (AMX + MTZ) resulted in greater clinical attachment level gain and probing pocket depth reduction compared with mechanical debridement alone. Jones and Hoyle found insufficient evidence to support routine adjunctive azithromycin, owing to low certainty and heterogeneity among studies. Discussion and Conclusions: Adjunctive systemic antibiotics, particularly AMX + MTZ, may provide additional clinical benefits in severe periodontitis. However, the limited number of systematic reviews, substantial overlaps, and uncertainty surrounding some antibiotic protocols limit the strength of the evidence. Current findings support selective rather than routine antibiotic use, guided by individual patient and disease characteristics and antimicrobial stewardship. Future studies should emphasize long-term outcomes and consistently distinguish Stage III from Stage IV periodontitis.

Background/Objectives: The application of organic soil amendments constitutes an important source of antibiotic residues in agricultural soils. However, studies have mainly focused on a few parent tetracyclines, whereas information on their transformation products (TPs) in organic amendments remains scarce. Therefore, this study assessed the occurrence and environmental risk of six tetracyclines and seven TPs in organic amendments. Methods: Processed livestock manures applied in the European Union (horse, poultry, and bovine manure), as well as fresh and treated sewage sludge, were analyzed using matrix solid-phase dispersion (MSPD) combined with online SPE-LC-MS/MS. Environmental risk was assessed using risk quotients (RQs) based on predicted environmental concentrations in soil (PECsoil) calculated from the maximum measured concentrations and predicted no-effect concentrations in soil (PNECsoil) obtained either directly from terrestrial ecotoxicity data or derived from aquatic ecotoxicity data using the equilibrium partitioning method and soil-water distribution coefficients. Results: Tetracyclines and their TPs were widely detected in both sample types, although concentrations varied according to matrix type and treatment. Overall, sludge showed higher detection frequencies and concentrations than manure. Doxycycline and tetracycline were the predominant parent compounds, reaching concentrations up to 2838 ng g[-1] dry weight (dw) in manure and 2892 ng g[-1] dw in sludge. Epimerized TPs were frequently detected and sometimes exceeded the concentrations of their parent compounds, especially epitetracycline and epioxytetracycline. Among the sludge types and treatment conditions investigated, anaerobically digested sludge showed the highest tetracycline concentrations, whereas the composted sludge sample presented the lowest concentrations. Individual RQs indicated insignificant to low ecotoxicological risk, whereas cumulative RQ (ΣRQ) values, used as conservative estimates of co-exposure to all evaluated tetracyclines and their TPs, fell within the medium-risk category for bovine manure and anaerobically digested sludge, with the composted sludge sample showing the lowest ΣRQ. Conclusions: These findings highlight the importance of including TPs in environmental monitoring and the differences in tetracycline occurrence and environmental risk to soil among the processed manure types and sludge treatment conditions investigated.

Infectious diarrhoea is a serious threat to farm animals, especially newborns. This study aimed to investigate in vitro selective growth-inhibitory effects of 15 Indian ethnoveterinary plant ethanolic extracts, 5 antibiotic growth promoters, 5 antibiotics, 15 phytochemicals and their synthetic analogues against 12 diarrhoea-associated and 12 probiotic bacteria of veterinary concern. Minimum inhibitory concentrations (MIC≥ 80) of crude extracts and chemicals were determined by the broth microdilution method and log value of the ratio of selective indices (SI) were calculated. Among the tested agents, 8-hydroxyquinoline and chloroxine (SI = 1.3 and 0.7) showed selective antibacterial effects comparable to those of ciprofloxacin, doxycycline, and tetracycline (SI = 1.1, 0.8 and 0.9), respectively. The extracts of Embelia ribes fruit and Punica granatum epicarp (SI = 0.3 and 0.2, respectively) produced weak selectivity, inhibiting the growth of diarrhoea-causing strains at lower concentrations than beneficial microbiota. An ultra-high-performance liquid chromatography equipped with a mass spectrometer and a high-resolution accurate mass (UHPLC/MS-HRAM) analysis of E. ribes extract, that showed a certain degree of selectivity, highest amongst tested species, quantified abundant embelin (256.7-327.6 µg/mg of dry extract), which could be responsible for its antibacterial activity. To our knowledge, this is the first study investigating the growth-inhibitory effects of E. ribes and embelin toward gastrointestinal bacteria of veterinary significance. In conclusion, our study identifies potential growth-inhibitory activities of 8-hydroxyquinoline and E. ribes extract, supporting further investigation of their potential relevance for veterinary antidiarrhoeal research. However, further studies are necessary to explore their applications in farm animals.

The implementation of HIV pre-exposure prophylaxis and doxycycline post-exposure prophylaxis may influence antimicrobial resistance in Neisseria gonorrhoeae among men who have sex with men (MSM). We examined trends in antimicrobial resistance among MSM, stratified by HIV and PrEP status, and assessed tetracycline resistance in relation to doxyPEP use. We included all MSM with an N. gonorrhoeae isolate at Melbourne Sexual Health Centre (MSHC) between 2015 and 2024. Resistance was defined using 2025 EUCAST breakpoints and epidemiological cutoff values. Trends were assessed using the Cochran-Armitage test, and tetracycline resistance was compared between doxyPEP users and non-users using the chi-square test. A total of 11,334 isolates from 6,611 MSM were included. Ceftriaxone resistance remained rare (3/11,334; 0.03%). The proportion of isolates with acquired azithromycin resistance mechanisms increased from 0.9% (8/940) in 2015 to 6.4% (102/1,584) in 2024 (Ptrend < 0.001), above the 5% level at which the WHO recommends that an antimicrobial no longer be used for empirical treatment. Ciprofloxacin resistance increased substantially, from 18.5% (170/920) to 55.2% (875/1,584) (Ptrend < 0.001), with significant trends in all three groups. Tetracycline resistance, assessable from 2019 onward, was consistently high (around 80%), with no significant trend. Among 1,020 isolates with known doxyPEP status, tetracycline resistance was significantly higher in doxyPEP users than in non-users (89.2% [107/120] vs 77.2% [695/900], P = 0.003). High-level tetracycline resistance (MIC > 8 mg/L) was also more common among doxyPEP users (54.2% vs 29.7%, P < 0.001). Ceftriaxone remains effective for empirical gonorrhea treatment among MSM in Melbourne. Rising the proportion of azithromycin non-wild-type raises concerns about its ongoing utility in dual therapy. Higher tetracycline resistance among doxyPEP users, particularly high-level tetracycline resistance, underscores the need to monitor for multidrug-resistant strains as doxyPEP use expands.IMPORTANCEAntimicrobial-resistant Neisseria gonorrhoeae is an urgent global health threat, and the expanding use of doxyPEP among MSM has raised concerns regarding its selective pressure on resistance. Linking a decade of individual-level clinical data to over 11,000 isolates from a major Australian sexual health clinic, we show that the proportion of azithromycin non-wild-type has risen to 6.4%, surpassing the 5% threshold at which the WHO recommends discontinuing an agent for empirical therapy. This finding raises concerns about the ongoing utility of dual ceftriaxone-azithromycin treatment. Furthermore, tetracycline resistance was significantly higher among doxyPEP users, with a disproportionate increase in high-level resistance (MIC >8 mg/L) that was associated with ciprofloxacin co-resistance. These findings provide timely real-world evidence to inform gonorrhea treatment guidelines and antimicrobial surveillance as doxyPEP implementation expands globally.

Hepatitis B virus (HBV) drives hepatocellular carcinoma in part through the activity of its X protein (HBx), yet the mechanisms by which HBx alters hepatocyte function remain incompletely understood. Progress has been limited by the lack of relevant human models that support controlled HBx expression in mature hepatocytes. To address this limitation, here, we use an improved hepatocyte-like organoid (HLO) platform that supports enhanced hepatocyte maturation to investigate HBx function in a differentiated hepatocyte context. Adult stem cell-derived HLOs were differentiated using an optimized protocol to generate hepatocyte-like cells with enhanced maturation and transcriptional similarity to primary liver tissue. HBx function was interrogated using both cognate promoter-driven expression and doxycycline-inducible systems across multiple donor-derived organoid lines. Transcriptomic, pathway, and single-cell imaging analyses were performed to assess the impact of HBx expression on hepatocytes. HBx expression consistently suppressed apoptosis-associated transcripts and reduced expression of core hepatocyte identity genes, including CYP3A4. Pathway analysis revealed downregulation of liver-specific functions, including metabolism, detoxification, complement, and coagulation. At the single-cell level, higher HBx expression was associated with reduced caspase 3/7 activation following apoptotic challenge and decreased hepatocyte marker expression. Functionally, the expression of HBx increased resistance to apoptosis and enhanced the ability of differentiated hepatocyte-like cells to revert to a proliferative state. HBx expression in differentiated human liver organoids reduces apoptosis and impairs hepatocyte identity, consistently across donors and expression systems. Together, these findings support a model in which HBx promotes a survival-permissive state of reduced hepatocyte differentiation. This state may contribute to early HBV-driven tumorigenesis. This HLO platform provides a relevant system to dissect HBV-host interactions and reveals a mechanism by which HBV may prime the liver for malignant transformation. IMPACT AND IMPLICATIONS: Understanding how HBV promotes hepatocellular carcinoma remains a critical challenge, partly due to the lack of physiologically relevant human derived model systems to study HBx function. Using a differentiated adult human liver organoid system, we show that HBx simultaneously suppresses apoptosis and reduces hepatocyte differentiation, providing a mechanistic framework for how HBV may prime hepatocytes for malignant transformation. These findings are particularly relevant for researchers studying HBV pathogenesis and liver cancer, as well as for clinicians aiming to better understand early disease progression. While further validation in more complex multicellular systems is needed, this platform can support the identification of HBx-targeted therapeutic strategies and guide the development of improved adult human derived models for virus-host interaction studies.

Human dirofilariasis is an underreported, vector-borne disease caused by the filarioid Dirofilaria immitis. A case of pulmonary dirofilariasis is reported in a woman from Costa Rica, manifesting respiratory distress, marked eosinophilia of 47%, and severe chylothorax. Pleural biopsies revealed a lymphoplasmacytic reaction with eosinophils, and bronchoalveolar fluid analysis revealed 28S and internal transcribed spacer 1 fragments that were 100% and 99.3% similar to D. immitis, respectively, and a coxA sequence identical to Wolbachia from D. immitis. Other infectious, autoimmune, and malignant causes were excluded through an extensive diagnostic workup. Initial treatment included methylprednisolone, albendazole, and a subsequent course of albendazole plus doxycycline; however, the chylothorax recurred. A biopsy of the left upper lobe confirmed a pleural lymphangiomatosis. The patient was discharged with continued treatment but was clinically stable. This case reveals the diagnostic complexity of filariasis and underlying lymphadenopathies coexisting in endemic regions and highlights the value of an integrated clinical, molecular, and histopathological approach.