Emerging Microbes & Infections最新文献

{"title":"Carbapenemase-producing <i>Klebsiella oxytoca</i> complex in France (2017-2023): molecular epidemiology integrated with virulence and biofilm phenotypes.","authors":"Inès Rezzoug, Océane Vanparis, Joséphine Poiraud, Subidsha Nandakumaran, Delphine Girlich, Léna Latour, Haniel Defoi, Cécile Emeraud, Agnès B Jousset, Aurelien Birer, Benoit Pilmis, Aymeric Jacquemin, Rémy A Bonnin, Laurent Dortet","doi":"10.1080/22221751.2026.2713319","DOIUrl":"10.1080/22221751.2026.2713319","url":null,"abstract":"<p><strong>Objectives: </strong>This study analysed a large French collection of carbapenemase-producing <i>Klebsiella oxytoca</i> complex (KoC) isolates (2017-2023) to map species and carbapenemase distributions and identify emerging lineages. We further investigated virulence traits to understand the epidemiological success of the dominant ST-2 lineage.</p><p><strong>Methods: </strong>We conducted a retrospective study of all KoC isolates received at the French National Reference Center for Antimicrobial Resistance from 2017 to 2023. Whole-genome sequencing was performed to assess species assignment, phylogeny, and sequence types. Virulence analyses included biofilm formation assays (on abiotic surfaces and eukaryotic cells) and <i>Galleria mellonella</i> infection models.</p><p><strong>Results: </strong><i>K. oxytoca sensu stricto</i> remained the most prevalent species among the KoC. OXA-48-like enzymes were the most common carbapenemases, and two major lineages predominated: ST-141 and, far ahead, ST-2. The success of ST-2 appears linked to its low virulence combined with a strong capacity for long-term persistence.</p><p><strong>Conclusion: </strong>Our findings confirm that <i>K. oxytoca</i> comprises a complex of distinct species whose taxonomy warrants continued revision. ST-2 circulates widely in France and, despite its limited virulence, shows remarkable persistence, making it an epidemiological important lineage.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2713319"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13508490/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148663555","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A rapid detection of Nipah virus nucleocapsid antigen in inactivated viral cultures.","authors":"Gyeongmu Choi, Jung Kim, Hansaem Lee, Pyeonghwa Jeon, Jae Hun Jeong, Yujin Jung, Jin Seo Park, Mi Kyoung Lee, Kyung-Chang Kim, Joo-Yeon Lee, Ha Yeon Park, Changwoo Park, Seil Kim, Hana Yi, Jihwan Song, Hyun-Joo Kim, Jong-Hwan Lee, Hong Gi Kim","doi":"10.1080/22221751.2026.2716499","DOIUrl":"10.1080/22221751.2026.2716499","url":null,"abstract":"<p><p>Nipah virus (NiV) is a highly lethal zoonotic henipavirus that causes severe respiratory disease and encephalitis and remains a priority pathogen for outbreak preparedness. Timely diagnosis is essential for containment, yet nucleic acid-based testing is often impractical in outbreak-prone, resource-limited settings. Here, we report the development of a rapid antigen test for NiV based on a cellulose nanobead (CNB)-enhanced lateral flow immunoassay (LFIA) targeting the viral nucleocapsid protein (NP). Anti-NiV NP antibodies were isolated from human synthetic phage-display libraries (scFv and Fab) by iterative bio-panning, reformatted as full-length IgGs, and screened using ELISA and immunofluorescence assays. Four lead antibodies were advanced to a sandwich-pairing matrix, which identified an optimized capture-detection configuration (mAb #5 as capture; mAb #2 as detection). Bio-layer interferometry confirmed nanomolar binding affinities for the selected antibodies (K_D = 39.50 nM for mAb #5; K_D = 26.36 nM for mAb #2). The finalized CNB-LFIA produced results within 15 min and achieved an analytical limit of detection of 1 ng/mL for recombinant NiV NP. The assay observed no cross-reactivity with SARS-CoV-2, SARS-CoV, MERS-CoV, or influenza A, whereas weak cross-reactivity for measles virus NP was observed only at a high antigen concentration (2 µg/mL). Using γ-irradiated cultured NiV quantified by RT-ddPCR, the LFIA detected viral load down to 4.1 × 10⁷ copies/mL (4.1 × 10⁶ copies/reaction) by a portable reader and 8.3 × 10⁷ copies/mL (8.3 × 10⁶ copies/reaction) by the naked eye. Collectively, this CNB-LFIA provides rapid, equipment-free NiV detection, supporting its potential utility for point-of-care diagnosis in resource-limited settings.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2716499"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13508541/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148683798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Early detection of pertussis related activity by wastewater surveillance of <i>Bordetella pertussis-</i> and <i>Bordetella parapertussis-</i>associated DNA concentrations in Osaka, Japan.","authors":"Koichiro Suzuki, Daisuke Motooka, Naomi Sakon, Kazunori Tomono","doi":"10.1080/22221751.2026.2717860","DOIUrl":"10.1080/22221751.2026.2717860","url":null,"abstract":"<p><p>Pertussis causes recurrent outbreaks worldwide. Clinical surveillance can be affected by healthcare-seeking behaviour, diagnostic practices, testing sensitivity, and reporting delays. This study evaluated the utility of wastewater surveillance (WWS) by quantifying <i>Bordetella-</i>associated <i>IS481</i> and <i>IS1001</i> DNA concentrations. Weekly wastewater samples were collected from four treatment plants in Osaka Prefecture, Japan, between April 2023 and July 2025, and <i>IS481</i>- and <i>IS1001</i>-associated DNA concentrations were measured using quantitative PCR. The putative <i>B. pertussis</i>-associated <i>IS481</i> DNA concentration increased markedly from 2024 onwards. Both threshold-based onset detection and cross-correlation analysis indicated that the wastewater <i>IS481</i> DNA concentration preceded reported cases by one to several months. In contrast, the <i>B. parapertussis-associated IS1001</i> DNA concentration exhibited a transient peak in 2023, which aligned with contemporaneous clinical reports but remained at low levels thereafter. These findings suggest that WWS may detect increases in pertussis-related activity earlier than clinical reporting and highlight the potential of WWS as a complementary tool to clinical surveillance for monitoring respiratory infectious diseases.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2717860"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13495037/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148705698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human pulmonary infection by <i>Brucella suis</i> vaccine strain S2 revealed by genome sequencing in China.","authors":"Kun Li, Xiaoyan Zhou, Lingling Tian, Leyu Wu, Dongri Piao, Yu Fan, Qingqing Xu, Hongyan Zhao, Pinggui Wang, Hai Jiang","doi":"10.1080/22221751.2026.2703398","DOIUrl":"10.1080/22221751.2026.2703398","url":null,"abstract":"<p><p>Brucellosis the most common zoonosis worldwide. For control and prevention of animal brucellosis, various <i>Brucella</i> vaccines have been developed. <i>Brucella suis</i> S2, a live attenuated vaccine strain proven safe to humans, is widely used in China. In this report, a 77-year-old woman with a history of coughing and expectoration for three years was diagnosed with brucellosis by next-generation sequencing. Subsequently, the <i>Brucella</i> isolate from this patient was determined to be <i>Brucella suis</i> vaccine strain S2 by phage lysis experiments, quantitative real-time PCR, and third-generation sequencing. Its nucleotide identity with vaccine strain S2 was as high as 99.9996%, and only 14 deletion/insertion mutations or single nucleotide polymorphisms were detected. A phylogenetic tree constructed based on the genome sequences revealed that this strain clustered together with strain S2. As the patient denied having a history of contact with animals or exposure to other known sources, an epidemiological investigation was carried out on environmental and food samples. The results did not reveal a clear infection source except for a <i>B. suis</i> S2-like strain (99.9939% identity to <i>B. suis</i> S2 strain) from a milk sample. Our study may provide the first reliable evidence that <i>B. suis</i> vaccine strain S2 has the potential to infect humans. While this needs further confirmation, it points to the need to survey the virulence and risks of vaccine strain S2 in humans. In addition, the route of transmission of vaccine strain S2 still needs to be clarified to prevent more cases of brucellosis originating from this strain.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2703398"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148591124","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dissemination of <i>bla</i>_KPC-3-harbouring <i>Klebsiella pneumoniae</i> across ST48 and ST628 in multiple healthcare facilities in the Republic of Korea.","authors":"Min Kyeong Kim, Kyeong Woo Noh, Eunkyung Shin, Jungsun Park, Ae Kyung Park, Seongjae Joo, Seo Hyun Lee, Da Eun Oh, Min Ji Kim, Min Hee Heo, Seung Yeon Jang, Junyoung Kim","doi":"10.1080/22221751.2026.2691352","DOIUrl":"10.1080/22221751.2026.2691352","url":null,"abstract":"<p><p><i>Klebsiella pneumoniae</i> carbapenemase-3 (KPC-3) remains rare in South Korea, where KPC-2 is the dominant carbapenemase, making the repeated detection of a concentrated <i>bla</i>_KPC-3 signal over five years notable. We performed genomic analyses of <i>bla</i>_KPC-3-harbouring <i>K. pneumoniae</i> from a regional healthcare network. Two chromosomally distinct lineages with concordant capsule loci (ST628/KL15 and ST48/KL62) presented multidrug-resistant phenotypes, and the virulence-associated loci were confined to ST48. Single-nucleotide polymorphism (SNP) analyses revealed near-clonal relatedness within lineages, with 0-38 pairwise SNPs among ST628 isolates and 8 SNPs between the two ST48 isolates. Core-genome multilocus sequence typing (cgMLST) supported this structure, as ST628 isolates were assigned to complex type 19149 with 0-7 allelic differences, and ST48 isolates were assigned to complex type 19150 with 5 allelic differences. These patterns support vertical spread via clonal expansion across multiple facilities. Despite substantial chromosomal separation, most isolates carried the same IncFII(K) plasmid backbone and <i>bla</i>_KPC-3, and they were nearly indistinguishable from a plasmid previously reported in South Korea. One isolate carried <i>bla</i>_KPC-3 on a distinct multireplicon IncFIB(K)/IncFII(K) plasmid, indicating that the signal was not confined to a single plasmid backbone. In both plasmids, <i>bla</i>_KPC-3 was embedded within Tn<i>4401b</i>. These findings indicate that a rare <i>bla</i>_KPC-3 genotype can persist regionally through sustained clonal dissemination and that cross-lineage linkage is compatible with past horizontal transfer involving a conserved plasmid. These findings underscore the need for subtype-resolved, regionally coordinated genomic surveillance in connected healthcare networks to detect uncommon carbapenemase variants early.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":"15 1","pages":"2691352"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13537060/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148873412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunogenicity and efficacy of a rabies-based vaccine against highly pathogenic influenza H5N1 virus.","authors":"Nir Paran, Christoph Wirblich, Randy Albrecht, Leila Zabihi-Diba, Alessandro Tarquinio, Drishya Kurup, Charalambos C Solomides, Adolfo García-Sastre, Matthias J Schnell","doi":"10.1080/22221751.2026.2620221","DOIUrl":"10.1080/22221751.2026.2620221","url":null,"abstract":"<p><p>The recent spillover of highly pathogenic influenza A/H5N1 (HPAI-H5N1) viruses to cattle, other mammals, and humans poses a major risk to animal and human health. Virus adaptation to new species highlights the need for effective vaccines for animals and humans. We recently developed a rabies virus-based H5 vaccine encoding the HPAI-H5 antigen and presenting it on the surface of the rabies virus particle. To test the immunogenicity and efficacy of the vaccine in eliciting systemic and mucosal immune response, we vaccinated mice intramuscularly or intranasally with either live or inactivated and adjuvanted vaccine. The vaccine elicited neutralizing antibodies against RABV and H5N1 Influenza virus and protected mice from a lethal challenge with PR8 recombinants reassorted with the HA of clade 1 (Viet Nam 1203) or clade 2.3.4.4b HPAI-H5N1 viruses, highlighting its potential use in mitigating the risk of HPAI-H5N1 pandemic.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2620221"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12875107/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146003338","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovery of the nucleoside analogue FMAU as a broad-spectrum anti-orthopoxvirus inhibitor in vitro and in vivo using a structure-guided hit expansion strategy.","authors":"Tingting Zheng, Yong Feng, Chang Xu, Arnaud Fondjo Kouam, Shiman Chen, Farkhod Eshboev, Yingxia Liu, Fuxiang Wang, Yang Yang, Jianxun Qi, Lifeng Fu","doi":"10.1080/22221751.2026.2735153","DOIUrl":"10.1080/22221751.2026.2735153","url":null,"abstract":"<p><p>Mpox remains a critical public health threat worldwide, and currently available therapeutics against orthopoxvirus infections remain very limited. In this study, we identified 2'-deoxy-2'-fluoro-5-methyl-1-β-D-arabinofuranosyluracil (FMAU) as a broad-spectrum anti-orthopoxvirus candidate through nucleos(t)ide analogue library screening combined with a structure-guided hit expansion strategy. In Vero cells, FMAU exerted potent in vitro antiviral activity against vaccinia virus (VACV) and monkeypox virus (MPXV), with EC₅₀ values of 1.99 μM and 30 nM, respectively, while exhibiting no apparent cytotoxicity, as reflected by a CC₅₀ value greater than 200 μM. In vivo mouse studies further showed that FMAU treatment provided full protection against lethal VACV and MPXV challenges, while significantly reducing viral loads in lung tissues. Importantly, oral FMAU administration retained potent therapeutic efficacy against MPXV in a mouse model of MPXV infection. Collectively, our findings identify FMAU as a promising oral anti-orthopoxvirus drug candidate and support its further preclinical evaluation.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2735153"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13600294/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148967145","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tropism of <i>Plasmodium malariae</i> for young red blood cells and the bone marrow.","authors":"Barbara Mouyama, Vivienne Fey, Dorenda J A Schoorl, Malika Serra-Hassoun, Fidelis Ambinintsoa, Barclaye Ngossanga, Leticia Hartmann, Ayola A Adegnika, Artur Scherf, Mario Recker, Rolf Fendel, Steffen Borrmann, Sylvie Garcia","doi":"10.1080/22221751.2026.2731507","DOIUrl":"10.1080/22221751.2026.2731507","url":null,"abstract":"<p><p>Fundamental aspects of the biology of the third most prevalent human malaria species <b><i>Plasmodium malariae</i></b>, including the basis for its capacity to establish chronic, low-density infections associated with debilitating quartan fevers, remain poorly characterized, hampering control efforts. Here, we combined ex vivo flowcytometric profiling of <i>P. malariae</i> isolates with infection studies in a recently developed humanized mouse model. We demonstrate a 5-fold higher tropism of <i>P. malariae</i> compared to <i>P. falciparum</i> for immature red blood cells, challenging a nearly century-old paradigm, and - for the first time - provide evidence for the parasite's capacity to home to and infect host bone marrow. Collectively, these findings reveal key life cycle features of this enigmatic and neglected human malaria parasite.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2731507"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13651831/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148905412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An interpretable machine learning model integrating mucosal and cellular immunity to predict SARS-CoV-2 reinfection: a multicenter cohort study.","authors":"Jinqiu Ran, Wen Zhang, Wanying Zhang, Hui Huang, Lejun Gong, Shengxia Yin, Yu Chen, Aofan Wang, Huijin Min, Liguo Zhu, Jinjin Chu, Yuanfang Qin, Rui Huang, Chao Wu, Yuxin Chen","doi":"10.1080/22221751.2026.2746253","DOIUrl":"10.1080/22221751.2026.2746253","url":null,"abstract":"<p><p>SARS-CoV-2 reinfection remains a significant public health challenge that requires accurate risk assessment across populations. Using prospectively collected data from two independent cohorts, we developed and externally validated an interpretable machine learning model for predicting symptomatic SARS-CoV-2 reinfection. The cohort comprised 1,156 adults from Wuxi and Nantong, China, with a median follow-up duration of 10 months. Reinfection occurred in 128 participants (11.1%). The model integrated eighteen clinical and immune features, and ultimately identified spike-specific nasal secretory immunoglobulin A (sIgA) and spike-specific CD4⁺ central memory T cells (T_CM) as key predictors. Developed using the extreme gradient boosting (XGB) algorithm, the prediction model achieved an AUC of 0.835 in the internal validation cohort and demonstrated strong discrimination in the external validation cohort. SHAP analysis revealed that higher nasal spike-specific sIgA was associated with lower risk, while spike-specific CD4⁺ T_CM cells exhibited a non-linear association with reinfection. This interpretable immune-based model provides a framework for immune-informed risk stratification, enabling better prediction and management of SARS-CoV-2 reinfection risk in settings with heterogeneous population immunity.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":"15 1","pages":"2746253"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13700530/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149237286","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assembling the puzzle of antimicrobial resistance in staphylococcal biofilms.","authors":"Thuy Nguyen, David McGiffin, Bin Lou, Yao Sun, Changrui Qian, Xenia Kostoulias, Wenhong Zhang, Anton Y Peleg, Yue Qu","doi":"10.1080/22221751.2026.2627073","DOIUrl":"10.1080/22221751.2026.2627073","url":null,"abstract":"<p><p>Multiple mechanisms underpinning biofilm antimicrobial resistance (AMR) have been studied individually. This study aimed to integrate these mechanisms, to understand their contributions to staphylococcal biofilm AMR, as a part of a whole, and to elucidate key hurdles hindering effective biofilm eradication by antimicrobial agents. Nine antibiotics were selected against microplate-based biofilms formed by <i>Staphylococcus aureus</i> ATCC 25923 and <i>Staphylococcus epidermidis</i> RP62A. Four mechanisms, including repressed bacterial metabolism, the barrier effect of the biofilm extracellular polymeric substances (EPS) matrix, the acidic inner-biofilm pH, and inoculum effects associated with high-cell-density biofilm growth were studied. The impact of individual mechanism on biofilm AMR was quantitated by determining the fold increase of concentration that allows antibiotics to overcome the mechanism. Antibiotic concentrations were then incrementally increased from minimum bactericidal concentration (MBC) to sequentially address all four mechanisms, ultimately aiming to kill at least 99.9% of biofilm cells. A simplified method was developed to evaluate the dependence of antibiotics on bacterial metabolic states for the lethality. Gentamicin, tobramycin and ciprofloxacin at 1024 µg/mL overcame all four mechanisms and successfully killed <i>S. aureus</i> ATCC 25923 biofilms by at least 3 log units. Ciprofloxacin at 1024 µg/mL effectively killed <i>S. epidermidis</i> RP62A biofilms. The contribution of each mechanism to biofilm AMR was strain- and drug-dependent, with low-cell metabolism being the most important factor. This study underscores the individual contributions of each mechanism to staphylococcal biofilm AMR and highlights the necessity of targeting all four mechanisms to achieve effective biofilm eradication.</p>","PeriodicalId":11602,"journal":{"name":"Emerging Microbes & Infections","volume":" ","pages":"2627073"},"PeriodicalIF":7.0,"publicationDate":"2026-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12903938/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146112567","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}