Microbial pathogenesis最新文献

{"title":"First comprehensive genome analysis and antimicrobial resistance profile of Clostridium innocuum from a domestic cat","authors":"Doğancan Yarım,&nbsp;Seçil Abay","doi":"10.1016/j.micpath.2025.108076","DOIUrl":"10.1016/j.micpath.2025.108076","url":null,"abstract":"<div><div>This study aimed to investigate the molecular and phenotypic properties of a <em>C. innocuum</em> isolate from a healthy cat, which was previously misidentified as <em>C. difficile</em> by molecular testing (<em>tpi</em>-PCR). To resolve the misidentification and obtain more information on the isolate at the genetic level, whole genome sequencing (WGS) on the long-read sequencing platform Oxford Nanopore PromethION was performed. WGS data were analysed to identify virulence factors, antibiotic resistance, and hypothetical prophage genes. The isolate was identified as <em>C. innocuum</em> as a result of WGS. While several virulence genes, including <em>pilM</em>, <em>yabQ</em>, <em>HrcA</em>, and <em>CptIN</em> family toxins, were detected, toxin-coding genes in other <em>Clostridium</em> species were not detected. Four prophage regions were detected; three appeared as intact phage sites, while one was incomplete. The isolate was susceptible to ampicillin, clindamycin, vancomycin, and metronidazole but was resistant to meropenem using the E-test method. The genome also contained resistance genes such as <em>tet(M), aph(2″)-Ia,</em> and <em>VanT, VanW, VanY, VanG,</em> and <em>VanZ</em>. Genetic similarities between our isolate and those obtained from global human-derived metagenome-associated genomes (MAGs) and non-MAG genomes from healthy humans and animals were illustrated through WGS-SNP-based phylogenetic analyses.</div><div>This study provides insights into the phenotypic and genotypic characteristics of <em>C. innocuum</em>, an emerging pathogen. Taken together, the isolation of this agent-possessing intrinsic resistance mechanisms and emerging after the treatment of <em>C. difficile</em> infections from a domestic cat highlights its potential threat to public health. Nevertheless, many clinical questions remain unanswered, underscoring the need for further research to deepen our understanding of this pathogen and to develop effective control and treatment strategies.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108076"},"PeriodicalIF":3.5,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145206753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to \"Feilike mixture targeting the STAT1-NLRP3 axis represses the pyroptosis of epithelial cells to alleviate Mycoplasma pneumoniae pneumonia\" [Microb. Pathogenesis. 206 (2025) 107754].","authors":"Chen Huang, Yanbin Fu","doi":"10.1016/j.micpath.2025.108066","DOIUrl":"https://doi.org/10.1016/j.micpath.2025.108066","url":null,"abstract":"","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":" ","pages":"108066"},"PeriodicalIF":3.5,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145192028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunological insights and vaccine advances against apicomplexan parasites: Emerging concepts and innovations","authors":"Ayed Alshammari","doi":"10.1016/j.micpath.2025.108074","DOIUrl":"10.1016/j.micpath.2025.108074","url":null,"abstract":"<div><div>The apicomplexan parasites are globally considered as the major cause of numerous infectious diseases in humans and animals. Apicomplexan parasites include <em>Plasmodium</em>, <em>Toxoplasma gondii</em>, <em>Cryptosporidium</em>, <em>Eimeria</em> and <em>Babesia</em>. The rise in the drug resistance have made the traditional control measures, such as chemotherapy and vector management, inadequate against them. These are the intracellular infectious agent and possess complex life cycles, antigenic variability, and immune evasion abilities. These different abilities hinder the development of vaccines against them. Hence, there is urgent need for development of effective vaccines by novel measures. However, notable progress has been made in past years due to the advancements in immunology, molecular biology, and biotechnology. Different types of vaccines including subunit vaccines have been developed and have demonstrated favorable efficiency. In the meantime, live-attenuated vaccines (LAV) continue to provide protection in animals. Apart from that, there are different innovations like CRISPR/Cas9 gene editing that have enabled the creation of genetically attenuated strains for <em>T. gondii</em> and <em>Eimeria</em>. These attenuated strains are used for the development of vaccines. Furthermore, mRNA vaccine technology, which was successfully utilized during the COVID-19 pandemic, is now being used against parasitic infections. It is now offering fast and rapid development along with vigorous cellular immunity. The use of nanoparticles and novel adjuvants such as TLR agonists and saponins has improved the stability and effectiveness of vaccines. Approaches like mucosal delivery, especially for enteric parasites such as <em>Cryptosporidium</em> and <em>Eimeria</em>, is achieving attention for their ability to provide the localized protection. In spite of these advancements some challenges still persist. Antigenic diversity, short-lived immunity, regulatory barriers, and limited funding need to be addressed. Some of the emerging technologies including systems vaccinology, reverse vaccinology, and vectored delivery platforms, are paving the way for more targeted and effective vaccination. There is need for concerted effort incorporating multidisciplinary research, One Health integration, and scalable manufacturing methodologies for effective translation of these scientific innovations into solutions. By harnessing these emerging technologies within a One Health framework, the next generation of vaccines has the potential to transform the management of apicomplexan diseases worldwide.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108074"},"PeriodicalIF":3.5,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145206745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chlamydia pneumoniae invades into vascular smooth muscle cells through the CXCR4-β-arrestin 2 pathway via TLR2/CXCR4 crosstalk","authors":"Yuke Zhang ,&nbsp;Lijun Zhang ,&nbsp;Yi Li ,&nbsp;Shuang Pan ,&nbsp;Guangyan Wang ,&nbsp;Qi Zhang ,&nbsp;Lijun Zhang","doi":"10.1016/j.micpath.2025.108067","DOIUrl":"10.1016/j.micpath.2025.108067","url":null,"abstract":"<div><div>Vascular smooth muscle cell (VSMC) dysfunction plays an important role in the pathogenesis of atherosclerosis. <em>Chlamydia pneumoniae</em> (<em>C. pneumoniae</em>) has been shown to infect and grow in VSMCs, and causes VSMC dysfunction, thereby promoting atherosclerosis development. However, it is an enigma how <em>C. pneumoniae</em> invades into VSMCs. In this study, we explored the specific mechanism of <em>C. pneumoniae</em> invading into VSMCs, and found that TLR2 expression and CXCR4 phosphorylation level were elevated after <em>C. pneumoniae</em> infection, and the translocation of TLR2 and CXCR4 to the membrane were increased at 15 min postinfection and reached the peak at 60 min, and then decreased at 180 min. The interaction between TLR2 and CXCR4 was enhanced in this process by co-immunoprecipitation and proximity ligation assays (PLA). The results from immunofluorescence staining demonstrated that <em>C. pneumoniae</em> invasion rate was dramatically decreased after knockdown of both TLR2 and CXCR4 compared with silencing of either TLR2 or CXCR4. Moreover, during <em>C. pneumoniae</em> invasion into VSMCs, the interaction between CXCR4 and β-arrestin2 was enhanced, and mutation of CXCR4 Ser339 resulted in the lack of β-arrestin2 recruitment. After TLR2 knockdown by siRNA, CXCR4 phosphorylation was decreased, and the translocation of CXCR4 to the membrane was abrogated, and CXCR4 was unable to recruit β-arrestin2 during <em>C. pneumoniae</em> invasion into VSMCs. In conclusion, <em>C. pneumoniae</em> invades into VSMCs through the CXCR4-β-arrestin2 pathway via TLR2/CXCR4 crosstalk, providing the first evidence for TLR2/CXCR4 interaction as well as receptor-mediated intracellular signaling that is exploited for <em>C. pneumoniae</em> invasion.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108067"},"PeriodicalIF":3.5,"publicationDate":"2025-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145192070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An on-site detection assay for screening of sendai virus by using palm-sized handheld system based on the reverse transcription multienzyme isothermal rapid amplification","authors":"You Jinwei ,&nbsp;Hong Jun ,&nbsp;Liang Lei ,&nbsp;Lu Yang ,&nbsp;Dong Min ,&nbsp;Ma Chang ,&nbsp;Zhang Xuliang ,&nbsp;Feng Xiaobo ,&nbsp;Chen Hui","doi":"10.1016/j.micpath.2025.108064","DOIUrl":"10.1016/j.micpath.2025.108064","url":null,"abstract":"<div><div>Sendai virus (SV) is the required inspection item for specific-pathogen-free grade rabbits and clean grade mice in the China National standard (GB14922.2–2011). Health monitoring should be performed regularly to avoid microbiological interference and obtain reliable experimental results. Reverse transcription multienzyme isothermal rapid amplification (RT-MIRA) technology allows for a high degree of specificity and sensitivity in pathogen detection. The aim of this study was to develop an on-site RT-MIRA assay with a handheld digitally thermostatic device to permit rapid visualization SV detection with naked eyes or displaying on phones connected via Bluetooth. SV-specific primers were targeted at the well-conserved L gene. The assay obtained results under isothermal conditions at 39 °C for 20 min. And the 95 % limit of detection was 33.5 copies/μL of SV RNA standards. Absence of non-specific amplification was confirmed by other common murine viruses. A nucleic acid releasing agent was used to lyse and release SV RNA within 5 min. Diagnostic performance of RT-MIRA on RNA extracted from 80 mice biological samples was compared with the reference method RT-PCR. SV was detected in 29 of 30 artificial positive samples and 0 of 50 clinical samples. The obtained results statistical analysis demonstrated that a good concordance was found between on-site RT-MIRA and the reference method RT-PCR (Kappa = 0.949). The on-site RT-MIRA assay exhibited high sensitivity and specificity, and the total time from sample collection to final result was approximately 30 min. We introduced a novel amplification technology and described an easy and efficient RT-MIRA assay for SV detection. This assay could be applied for simple on-site visual detection of SV.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108064"},"PeriodicalIF":3.5,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145182094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rhizopus arrhizus infection induces histopathological lung injury and alters immune and membrane-associated pathways in a murine model","authors":"Mingquan Qiu ,&nbsp;Zishi Liu ,&nbsp;Li Wang ,&nbsp;Yong Wei","doi":"10.1016/j.micpath.2025.108058","DOIUrl":"10.1016/j.micpath.2025.108058","url":null,"abstract":"<div><div>As a major pathogenic fungus of the Rhizopodaceae family, <em>Rhizopus arrhizus</em> can infect humans and lead to severe outcomes, including life-threatening systemic infections. In this study, a fungal strain designated XML01 was isolated from a goat specimen. Following cultivation on potato dextrose agar (PDA), the isolate was identified as <em>Rhizopus arrhizus</em> through lactophenol cotton blue staining, scanning electron microscopy, and ITS gene sequencing. To investigate the pulmonary pathogenicity and mechanistic underpinnings of this strain, a murine model was established and analyzed using histopathological techniques (HE and Gomori's methenamine silver staining), RT-qPCR, and transcriptome sequencing. Infection with <em>R. arrhizus</em> resulted in significant pulmonary damage, characterized by alveolar wall thickening, severe hemorrhage, inflammatory cell infiltration, and interstitial hyperplasia. Transcriptome analysis revealed 318 significantly differentially expressed genes, predominantly enriched in pathways related to ciliary motility, cGMP-PKG signaling, and calcium homeostasis—indicating profound disruption of normal lung function. Key downregulated genes were associated with the IL-17 and B-cell receptor signaling pathways. Notably, <em>SftpC</em> and <em>GSN</em> were significantly upregulated, while <em>Fth1</em>, <em>Scgb3a1</em>, and <em>Scgb1a1</em> were downregulated, findings that were consistent with RT-qPCR validation. Collectively, this work provides a novel and highly reproducible animal model that deepens the understanding of pathogenesis and offers a valuable tool for the development of new therapies for mucormycosis.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108058"},"PeriodicalIF":3.5,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176508","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combining experimental and AI approaches to identify antifungal compounds of Origanum vulgare and Mentha pulegium essential oils targeting Candida albicans","authors":"Amel Benmessaoud ,&nbsp;Wassim Yezli ,&nbsp;Mohamed Ouassini Bensaid ,&nbsp;Dou El Kefel Mansouri","doi":"10.1016/j.micpath.2025.108039","DOIUrl":"10.1016/j.micpath.2025.108039","url":null,"abstract":"<div><div>The growing issue of antifungal resistance and the persistence of microbial biofilms underscores the need for novel therapeutic strategies. This study presents an Artificial Intelligence (AI)-based framework to predict the antifungal and antibiofilm effectiveness of essential oils from <em>Origanum vulgare</em> L. and <em>Mentha pulegium</em> L., both individually and in combination with nystatin, against <em>Candida albicans</em>. Three machine learning models—linear regression, AdaBoost, and Random Forest—were employed to predict the Minimum Inhibitory Concentrations (MICs) and antibiofilm activity. Furthermore, a feature selection algorithm was used to identify the most influential bioactive compounds, with thymol and pulegone emerging as key predictors in the models. Models'performance was assessed using Root Mean Square Error (RMSE) and Mean Absolute Error (MAE), and validated through experimental methods, including disk diffusion, microdilution method, antibiofilm efficacy, and the checkerboard method to assess synergistic effects with nystatin. Experimental results closely corroborated the predictions of the proposed AI models, further substantiating the significant antifungal and antibiofilm properties of these essential oils. GC-MS/GC-FID analysis for the identification of active compounds revealed that thymol (53.23 %) and pulegone (70.50 %) were the major constituents of <em>O. vulgare</em> and <em>M. pulegium</em>, respectively. Moreover, the combination of nystatin with <em>O. vulgare</em> essential oil showed synergistic activity against both planktonic and sessile forms of <em>C. albicans</em>. These findings validate the predictive power of AI models, highlighting the potential of integrating data science with biology to optimize personalized antifungal treatments and discover new compounds with high therapeutic potential in combating resistant fungal infections.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108039"},"PeriodicalIF":3.5,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145156262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hinokitiol disrupts iron homeostasis and reduces virulence in Acinetobacter baumannii","authors":"Horng-Ren Lo ,&nbsp;Yun-En Wu ,&nbsp;Chung-Cheng Lo ,&nbsp;Tzu-Hui Yeh ,&nbsp;Rui-Yu Yeh ,&nbsp;Wen-Yu Lin","doi":"10.1016/j.micpath.2025.108062","DOIUrl":"10.1016/j.micpath.2025.108062","url":null,"abstract":"<div><h3>Background</h3><div>Multidrug-resistant gram-negative bacterial infections pose a serious threat to human health. Given that iron levels affect bacterial pathogenesis, iron homeostasis is a promising target for antimicrobial therapies. We investigated the effects and mechanisms of action of hinokitiol against <em>Acinetobacter baumannii</em>, a notorious opportunistic pathogen that causes nosocomial infections worldwide.</div></div><div><h3>Main methods</h3><div>The minimum inhibitory concentration was determined using the broth microdilution method. The inhibition of <em>A. baumannii</em> isolates was determined by growth kinetics analysis and viable counts with or without iron supplementation following hinokitiol exposure. The expression status of acinetobactin-related genes and a motility-related gene was evaluated using RT-qPCR. The bioenergetics-related activity of hinokitiol-treated bacteria was assessed by determining the intracellular ATP levels and bacterial motility. The fluorescent probe 2ʹ,7′-dichlorodihydrofluorescein diacetate was used to detect intracellular reactive oxygen species.</div></div><div><h3>Results</h3><div>Hinokitiol decreased the number of viable bacteria in time- and concentration-dependent manners against <em>A. baumannii</em>. Hinokitiol-treated bacteria exhibited reduced intracellular iron levels and elevated expression of acinetobactin-related genes compared with untreated cells. Intracellular ATP levels, motility, and virulence-related characteristics were reduced; however, intracellular reactive oxygen species levels increased in <em>A. baumannii</em> following hinokitiol exposure. More survival nematodes were recorded for the <em>Caenorhabditis elegans</em> infection model using hinokitiol.</div></div><div><h3>Significance</h3><div>Hinokitiol affects bacterial iron homeostasis. Impaired bioenergetics-related activity and induced oxidative stress after hinokitiol treatment may contribute to the reduced viability of <em>A. baumannii</em>. Hinokitiol reduced bacterial adhesion, invasion, and cytotoxicity, which may be beneficial for treating <em>A. baumannii</em> infections.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108062"},"PeriodicalIF":3.5,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145156348","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improved biodegradation of 17β-estradiol by indigenous microorganisms and assessment of its ecotoxicity","authors":"Karpaga Valli Arumugam,&nbsp;Venkat Kumar Shanmugam","doi":"10.1016/j.micpath.2025.108063","DOIUrl":"10.1016/j.micpath.2025.108063","url":null,"abstract":"<div><div>Estrogen is a steroid hormone that damages aquatic animal ecology and human health because it contains two hydroxyl groups in its structure. Additionally, the environment is impacted by even a tiny ng/L. This study aims to isolate the organisms from drug-contaminated soil and to degrade 17β-estradiol from the environment. The isolated strains selected for degradation purposes were VITVK1, VITVK2, and VITVK3. The three strains were identified as rod-shaped bacteria the strains are <em>Bacillus infantis, Achromobacter xyloxidans,</em> and <em>Sphingomonas</em> sp. The genebank accession number was Bacillus infantis - PQ144088, Achromobacter xyloxidans - PQ155434, and Sphingomonas sp. - PQ144536. These strains are crucial to the current investigation; with Achromobacter xyloxidans achieving the highest degradation percentage of 17β-estradiol at 84 %, <em>Sphingomonas</em> sp. achieving 72 %, and <em>Bacillus infantis</em> achieving the lowest at 54 %. The maximum degraded bacteria sample was processed for optimization studies and analysed for SEM it was identified that degradation shows the changes in cell surface or the creation of extracellular materials are responsible for the degradation process of <em>Achromobacter xylosoxidans,</em> and GC-MS predicts the peak reduction of 0^th hr-12th hr. Helminth toxicity of <em>Eisenia fetida</em> and phytotoxicity of <em>Eleusine coracana</em> of 17β-estradiol shows high toxicity in the untreated sample. The study emphasizes how indigenous microbes can be used to bioremediate areas contaminated with 17β-estradiol and help degrade the endocrine-disrupting chemicals.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108063"},"PeriodicalIF":3.5,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145149987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Seed transmission and genome characterization of tobacco streak virus in marigold: Development of qPCR and RT-LAMP assays for host and vector detection","authors":"Niresh Kumar S ,&nbsp;G.S. Madhu ,&nbsp;V. Venkataravanappa","doi":"10.1016/j.micpath.2025.108054","DOIUrl":"10.1016/j.micpath.2025.108054","url":null,"abstract":"<div><div>Marigold plants (30 samples) exhibiting virus-like symptoms were collected from diverse agro-climatic regions of Karnataka, Maharashtra, and Tamil Nadu to identify the causal agent. Initial serological assays indicated a positive reaction to tobacco streak virus (TSV). Infectivity was confirmed by mechanically inoculating symptomatic leaf extracts onto cowpea indicator plants (cv. C-152), which developed characteristic symptoms, demonstrating biological activity of the virus. For molecular confirmation, total RNA was extracted from eight representative plant samples (seven marigold and one cowpea), two seed samples, and five thrips collected from infected fields. RT-PCR with coat protein (CP) gene-specific primers consistently amplified a ∼700 bp fragment from marigold, cowpea, and thrips samples. Sequencing of cloned amplicons revealed &gt;97 % nucleotide identity with Indian TSV isolates infecting okra, cotton, groundnut, sunflower, and cucumber. Given the frequency of TSV incidence and the extent of marigold cultivation, one representative isolate was selected for complete genome sequencing. Using segment-specific primers, RNA1, RNA2, and RNA3 were amplified, cloned, and sequenced. Comparative genome analysis showed &gt;97 % identity with Indian isolates and with TSV isolates from squash and pumpkin in the United States, suggesting cross-host adaptation. Sequencing of the mitochondrial cytochrome oxidase I (<em>mt</em>COI) gene identified <em>Thrips tabaci</em> as the vector transmitting TSV. Furthermore, quantitative PCR (qPCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays were developed and validated for rapid and sensitive TSV detection in marigold tissues and thrips. This study reports the first complete genome sequence of marigold-associated TSV isolate, confirming its dual transmission via seeds and thrips, and providing critical insights into its molecular epidemiology and threat to ornamental crops in India.</div></div>","PeriodicalId":18599,"journal":{"name":"Microbial pathogenesis","volume":"209 ","pages":"Article 108054"},"PeriodicalIF":3.5,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145120017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}