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Trans-cinnamaldehyde (TC), a plant-derived antimicrobial, was examined against Edwardsiella ictaluri, including how the bacterium adapts after prolonged exposure and whether TC-adapted strains can act as vaccine candidates. TC inhibited E. ictaluri at an MIC of 120 µg/mL; subinhibitory dosing (16 µg/mL) enabled long-term adaptation. Adapted strains showed reduced TC and florfenicol susceptibility, increased SXT susceptibility, attenuated virulence, and partial to high survival after vaccination challenge. Proteomics and comparative genomics indicated efflux pump and stress protein upregulation, metabolic reprogramming, and shared mutations in membrane and regulatory pathways during adaptation.",{"@graph":69,"@context":123},[70,84,106],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/investigation-mechanisms-of-action-and-resistance-of-edwardsiella-ictaluri-to-trans-cinnamaldehyde/455378/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/investigation-mechanisms-of-action-and-resistance-of-edwardsiella-ictaluri-to-trans-cinnamaldehyde/455378.png","ImageObject",300,407,{"name":92,"@type":93},"acoobca","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-30","2026-09-30",true,{"@type":102,"interactionType":103,"userInteractionCount":105},"InteractionCounter",{"@type":104},"ViewAction",12,{"@type":107,"mainEntity":108},"FAQPage",[109,115,119],{"name":110,"@type":111,"acceptedAnswer":112},"What is the main threat addressed by the study in aquaculture?","Question",{"text":113,"@type":114},"Multidrug-resistant (MDR) bacterial pathogens threaten food safety and public health by enabling resistance transfer through the food chain.","Answer",{"name":116,"@type":111,"acceptedAnswer":117},"How was trans-cinnamaldehyde (TC) used to study adaptation in Edwardsiella ictaluri?",{"text":118,"@type":114},"TC antimicrobial activity was quantified by MIC testing, and long-term adaptation was induced using a subinhibitory concentration (16 µg/mL) with serial passages for 30 and 60 days.",{"name":120,"@type":111,"acceptedAnswer":121},"What evidence supports the potential of TC-adapted strains as vaccines?",{"text":122,"@type":114},"Vaccination with D30-TC and D60-TC strains significantly protected catfish against wild-type challenge, with reported survival increases compared with controls.","https://schema.org",{"og:url":83,"og:type":125,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":127,"canonical":83},"index,follow",{"doc_id":129,"site_id":62},455378,1790789991,{"code":4,"msg":5,"data":132},{"doc_id":129,"user_id":133,"nickname":92,"user_avatar":134,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":135,"file_id":136,"file_url":137,"file_type":138,"file_size":139,"view_count":105,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":140,"language":141,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":142,"faqs":143,"seo_title":144,"seo_description":67,"update_tm":145,"read_time":146,"preview_page_count":4},3985747870733,"https://ap-avatar.wpscdn.com/davatar_a8503ba1806abce46bf441b54a3ca4cd","OPEN ACCESS  \nCitation: Abdelhamed H, Riman MM, Arick MA, Dawood AS, Gomaa B, Chowdhury M, et al.(2026) Investigation mechanisms of action and resistance of Edwardsiella ictaluri to transcinnamaldehyde. PLoS One 21(1): e0340053 .  \n[https://doi.org/10.1371/journal.pone.0340053](https://doi.org/10.1371/journal.pone.0340053)  \n[Editor:](Editor: Nermeen M)[ Nermeen M](Editor: Nermeen M). Abu-Elala, CU Faculty of Veterinary Medicine: Cairo University Faculty of Veterinary Medicine, EGYPT  \nReceived: August 14, 2025  \nAccepted: December 16, 2025  \nPublished: January 7, 2026  \nCopyright: © 2026 Abdelhamed et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.  \nData availability statement: All relevant data are within the manuscript and its Supporting  Information files.  \nFunding: The research was funded by the United States Department of Agriculture (USDA) National Institute of Food and  \nRESEARCH ARTICLE  \nInvestigation mechanisms of action and resistance of Edwardsiella ictaluri to trans-cinnamaldehyde  \nHossam Abdelhamed1*, Munshi Mustafiz Riman1, Mark A. Arick II2, Ali Sobhy Dawood1, Basant Gomaa1, Monzur Chowdhury1, Larry A. Hanson1, Attila Karsi1  \n1 Department of Comparative Biomedical Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, Mississippi, United States of America, 2 Institute for Genomics, Biocomputing and Biotechnology, Mississippi State University, Mississippi State, Mississippi, United States of America  \n* [abdelhamed@cvm.msstate.edu](abdelhamed@cvm.msstate.edu)  \nAbstract  \nThe rise of multidrug-resistant (MDR) pathogens in aquaculture poses a significant threat to food safety and public health by facilitating the transfer of resistance across the food chain, underscoring the need for sustainable, non-antibiotic control measures. Trans-cinnamaldehyde (TC), a phytochemical with antimicrobial activity, is a promising alternative. While previous publications have established the antibacterial efficacy of TC against Edwardsiella ictaluri, the agent of enteric septicemia of catfish, its mechanism of action and the potential for bacterial adaptation after prolonged exposure remain undefined. Here, we addressed these gaps by investigating the antibacterial mechanism, the adaptive response of E. ictaluri to TC, and the vaccine potential of TC-adapted strains, with relevance to One-Health. The minimum inhibitory concentration (MIC) of TC against E. ictaluri was 120 µg/mL, whereas 16 µg/mL was not inhibitory and was used for long-term adaptation. Serial passages for 30 and 60 days generated D30-and D60-adapted strains. Disk diffusion assays indicated reduced susceptibility to TC and florfenicol but increased susceptibility to sulfamethoxazole/trimethoprim (SXT) in adapted strains. Virulence assays in catfish fingerlings (three tanks containing 10 fish per tank) demonstrated that D30-TC and D60-TC strains were highly attenuated. Moreover, vaccination with these strains conferred significant protection against wild-type challenge (63 .64% and 73.64% survival compared to 20% in controls) . Quantitative proteomics revealed the upregulation of multidrug efflux pumps and stress adaptation proteins and the downregulation of central metabolic pathways during TC exposure. Long-term adaptation was characterized by metabolic reprogramming, including enhanced energy and carbohydrate metabolism and suppression of virulence-associated secretion systems. Comparative genomics of D30-TC and D60-TC strains revealed shared mutations in genes related to membrane biosynthesis, energy metabolism, stress response, and regulatory pathways.  \nPLOS One | [https://doi.org/10.1371/journal.pone.0340053](https://doi.org/10.1371/journal.pone.0340053) January 7, 2026 1 / 27  \nAgriculture (NIFA) awarded number 2022- 67015-36339","cbCail5OEwmERBhi","https://ap.wps.com/l/cbCail5OEwmERBhi","pdf",1288773,27,"English","# Abstract\n# Introduction","[{\"question\":\"What is the main threat addressed by the study in aquaculture?\",\"answer\":\"Multidrug-resistant (MDR) bacterial pathogens threaten food safety and public health by enabling resistance transfer through the food chain.\"},{\"question\":\"How was trans-cinnamaldehyde (TC) used to study adaptation in Edwardsiella ictaluri?\",\"answer\":\"TC antimicrobial activity was quantified by MIC testing, and long-term adaptation was induced using a subinhibitory concentration (16 µg/mL) with serial passages for 30 and 60 days.\"},{\"question\":\"What evidence supports the potential of TC-adapted strains as vaccines?\",\"answer\":\"Vaccination with D30-TC and D60-TC strains significantly protected catfish against wild-type challenge, with reported survival increases compared with controls.\"}]","Investigation mechanisms of action and resistance of Edwardsiella ictaluri to trans-cinnamaldehyde | PDF",1790742731,68]