{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,13]],"date-time":"2026-06-13T16:43:11Z","timestamp":1781368991194,"version":"3.54.1"},"reference-count":60,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,4,24]],"date-time":"2025-04-24T00:00:00Z","timestamp":1745452800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Combating antimicrobial resistance is one of the most pressing public health challenges of our time. The rapid spread of resistant, zoonotic bacterial strains in livestock farming is increasingly raising concerns about the need to reduce antibiotic use. Because of this, there is an urgent need for safe and effective alternatives in animal husbandry.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>This study aimed to perform an <jats:italic>in vivo<\/jats:italic> the dose\u2013response analysis of fenugreek (<jats:italic>Trigonella foenum-graecum<\/jats:italic>), as a plant-based antibiotic alternative feed supplement in B\u00e1bolna Tetra-SL chicks (1\u201342\u202fdays old) with a 1:1 sex ratio. A total of 270 chicks were randomly assigned to 18 groups (15 birds per group) and subjected to six different treatment groups in three replicates: fenugreek at 1\u00d7, 10\u00d7, and 100\u00d7 doses, an antibiotic-treated group (enrofloxacin), a positive control group (infection only), and a negative control group (no infection or treatment). The infection was induced using mixed <jats:italic>Salmonella Enteritidis<\/jats:italic> and <jats:italic>Escherichia coli<\/jats:italic>, administered via gavage on days 3 and 4 of life. The birds were monitored for clinical symptoms, body weight, feed intake, and <jats:italic>Salmonella<\/jats:italic> shedding through cloacal swab samples. Statistical analyses included mixed-effect logistic regression for mortality, mixed-effect linear models for weight gain, two-way ANOVA for feed efficiency, and random effects continuation ratio models for <jats:italic>Salmonella<\/jats:italic> isolation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Significant interactions for Group:Day and Sex:Day in weight gain were identified (<jats:italic>p<\/jats:italic>\u202f&amp;lt;\u202f0.0001 for both). Additionally, the 1\u202f\u00d7\u202fdose group showed significantly reduced <jats:italic>Salmonella<\/jats:italic> shedding compared to the positive control group on day 33 (<jats:italic>p<\/jats:italic>\u202f=\u202f0.0031). The low-dose group (1\u00d7) demonstrated the most promising results, showing a 63% reduction in <jats:italic>Salmonella<\/jats:italic> shedding on day 10 and 31% on day 17. This group exhibited the fewest clinical symptoms, no diarrhea, and the lowest individual and specific feed intake up to day 24.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>The findings of this study suggest that low-dose fenugreek supplementation could be a viable strategy for reducing <jats:italic>Salmonella<\/jats:italic> shedding in poultry, potentially contributing to reduced antibiotic use in poultry farming and thus playing a role in the global effort to combat antimicrobial resistance. Future research will involve large-scale industrial trials and next-generation sequencing to evaluate the additive\u2019s impact on gut microbiota composition.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fvets.2025.1570387","type":"journal-article","created":{"date-parts":[[2025,4,24]],"date-time":"2025-04-24T05:30:17Z","timestamp":1745472617000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Dose\u2013response study of a fenugreek-based antibiotic alternative in B\u00e1bolna Tetra-SL chicks (1\u201342\u202fdays old) with mixed bacterial infections"],"prefix":"10.3389","volume":"12","author":[{"given":"\u00c1d\u00e1m","family":"Kerek","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1bel","family":"Szab\u00f3","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P\u00e9ter Ferenc","family":"Dobra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krisztina","family":"B\u00e1rdos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bettina","family":"Paszerbovics","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zs\u00f3fia","family":"Bata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vivi\u00e1na","family":"Moln\u00e1r-Nagy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1kos","family":"Jerzsele","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L\u00e1szl\u00f3","family":"\u00d3zsv\u00e1ri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,4,24]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"e51034","DOI":"10.15252\/embr.202051034","article-title":"Antibiotic resistance and persistence\u2014implications for human health and treatment perspectives","volume":"21","author":"Huemer","year":"2020","journal-title":"EMBO Rep"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"e0116523","DOI":"10.1371\/journal.pone.0116523","article-title":"Long-lasting effects of early-life antibiotic treatment and routine animal handling on gut microbiota composition and immune system in pigs","volume":"10","author":"Schokker","year":"2015","journal-title":"PLoS One"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"6644","DOI":"10.3382\/ps\/pez517","article-title":"The technical and economic impact of veterinary interventions aimed at reducing antimicrobial use on broiler farms","volume":"98","author":"Roskam","year":"2019","journal-title":"Poult Sci"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"1673","DOI":"10.3390\/antibiotics11111673","article-title":"Potential of aromatic plant-derived essential oils for the control of foodborne Bacteria and antibiotic resistance in animal production: a review","volume":"11","author":"Zhang","year":"2022","journal-title":"Antibiotics"},{"key":"ref5","first-page":"397","article-title":"The possibilities of antibiotic-free broiler-hen fattening, with special reference to the use of pre- and probiotics","volume":"142","author":"Ess\u0151sy","year":"2020","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref6","first-page":"281","article-title":"Usage of antibiotic alternatives in pig farming: literature review","volume":"143","author":"Kov\u00e1cs","year":"2021","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"181","DOI":"10.56385\/magyallorv.2024.03.181-191","article-title":"Antimicrobial peptides as new tools to combat infectious diseases","volume":"146","author":"Seb\u0151k","year":"2024","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"497","DOI":"10.56385\/magyallorv.2023.08.497-510","article-title":"Antimicrobial properties of essential oils \u2013 animal health aspects","volume":"145","author":"Kov\u00e1cs","year":"2023","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"706","DOI":"10.3390\/vetsci10120706","article-title":"Determining the in vivo efficacy of plant-based and probiotic-based antibiotic alternatives against mixed infection with Salmonella enterica and Escherichia coli in domestic chickens","volume":"10","author":"Kerek","year":"2023","journal-title":"Vet Sci"},{"key":"ref10","first-page":"77","article-title":"Effects of fermented wheat germ extract on artificial Salmonella Typhimurium infection in broiler chickens","volume":"142","author":"Jerzsele","year":"2020","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"461","DOI":"10.56385\/magyallorv.2023.08.461-475","article-title":"In vivo efficacy of different extracts of Propolis in broiler salmonellosis","volume":"145","author":"Olasz","year":"2023","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref12","first-page":"285","article-title":"Antibacterial efficiency of propolis \u2013 part 1","volume":"144","author":"Kerek","year":"2022","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref13","first-page":"691","article-title":"Antiprotozoal and antifungal efficiency of propolis \u2013 part 2","volume":"144","author":"Kerek","year":"2022","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"651","DOI":"10.56385\/magyallorv.2024.11.651-659","article-title":"Physiological effects of medium-chain fatty acids and triglycerides, and their potential use in poultry and swine nutrition: a literature review","volume":"146","author":"Het\u00e9nyi","year":"2024","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"419","DOI":"10.56385\/magyallorv.2023.07.419-438","article-title":"Antibacterial therapy based on pharmacokinetic\/pharmacodynamic models in small animal medicine-1. Literature review","volume":"145","author":"Mag","year":"2023","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"723","DOI":"10.56385\/magyallorv.2024.12.723-742","article-title":"Biosecurity situation of large-scale poultry farms in Hungary according to the databases of National Food Chain Safety Office Centre for disease control and biosecurity audit system of poultry product Board of Hungary in the period of 2021\u20132022","volume":"146","author":"Farkas","year":"2024","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"291","DOI":"10.56385\/magyallorv.2024.05.291-305","article-title":"Study report on the effect of a litter treatment product containing Bacillus licheniformis and zeolite in male fattening Turkey flock","volume":"146","author":"Kov\u00e1cs","year":"2024","journal-title":"Magyar \u00c1llatorvosok Lapja"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"863","DOI":"10.3390\/ijerph15050863","article-title":"Campylobacteriosis, salmonellosis, Yersiniosis, and Listeriosis as zoonotic foodborne diseases: a review","volume":"15","author":"Chlebicz","year":"2018","journal-title":"Int J Environ Res Public Health"},{"key":"ref19","doi-asserted-by":"publisher","DOI":"10.1128\/microbiolspec.AME-0011\u20132019","article-title":"Molecular epidemiology of infectious zoonotic and livestock diseases","volume":"8","author":"Gebreyes","year":"2020","journal-title":"Microbiol Spectr"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"407","DOI":"10.3390\/microorganisms8030407","article-title":"Salmonella virulence and immune escape","volume":"8","author":"Wang","year":"2020","journal-title":"Microorganisms"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.imlet.2017.07.006","article-title":"Salmonella infection: interplay between the bacteria and host immune system","volume":"190","author":"Kurtz","year":"2017","journal-title":"Immunol Lett"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"110","DOI":"10.1016\/j.cmi.2015.12.004","article-title":"Salmonellosis: the role of poultry meat","volume":"22","author":"Antunes","year":"2016","journal-title":"Clin Microbiol Infect"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"4273","DOI":"10.1128\/AEM.00598-11","article-title":"Population dynamics of Salmonella enterica serotypes in commercial egg and poultry production","volume":"77","author":"Foley","year":"2011","journal-title":"Appl Environ Microbiol"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"e0268231","DOI":"10.1371\/journal.pone.0268231","article-title":"The prevalence and concentration of Salmonella enterica in poultry litter in the southern United States","volume":"17","author":"Dunn","year":"2022","journal-title":"PLoS One"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"47","DOI":"10.3390\/biom10010047","article-title":"Anticancer plants: a review of the active phytochemicals, applications in animal models, and regulatory aspects","volume":"10","author":"Khan","year":"2020","journal-title":"Biomol Ther"},{"key":"ref26","first-page":"20","article-title":"Therapeutic applications of fenugreek","volume":"8","author":"Basch","year":"2003","journal-title":"Altern Med Rev"},{"key":"ref27","author":"Gaikwad","year":"2018"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"53","DOI":"10.30466\/vrf.2019.80961.2072","article-title":"The effect of different levels of fenugreek (Trigonella foenum-graecum L.) powder and extract on performance, egg quality, blood parameters and immune responses of laying hens in second production cycle","volume":"11","author":"Khalili Samani","year":"2020","journal-title":"Vet Res Forum"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"57","DOI":"10.3390\/vetsci11020057","article-title":"Modulation of immune response and Cecal microbiota by dietary fenugreek seeds in broilers","volume":"11","author":"Paneru","year":"2024","journal-title":"Vet Sci"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"11","DOI":"10.3390\/microorganisms5010011","article-title":"The food production environment and the development of antimicrobial resistance in human pathogens of animal origin","volume":"5","author":"Lekshmi","year":"2017","journal-title":"Microorganisms"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"e8856","DOI":"10.2903\/j.efsa.2024.8856","article-title":"EFSA panel on additives and products or substances used in animal feed (FEEDAP). Guidance on the assessment of the efficacy of feed additives","volume":"22","author":"Bampidis","year":"2024","journal-title":"EFSA J"},{"key":"ref32","author":"Rizopoulos","year":"2023"},{"key":"ref33","author":"Jos\u00e9","year":"2024"},{"key":"ref34","article-title":"Analysis of variance; designed experiments","volume-title":"Statistical Models in S","author":"Chambers","year":"1992"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.18637\/jss.v067.i01","article-title":"Fitting linear mixed-effects models using lme4","volume":"67","author":"Bates","year":"2015","journal-title":"J Stat Softw"},{"key":"ref36","author":"Chambers","year":"1992"},{"key":"ref37","doi-asserted-by":"publisher","first-page":"637","DOI":"10.29261\/pakvetj\/2024.215","article-title":"Impact of dietary Bacillus toyonensis M44 as an antibiotic alternative on growth, blood biochemical properties, immunity, gut microbiota, and meat quality of IR broilers","volume":"3","author":"Fatimah","year":"2024","journal-title":"Pak Vet J"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"4","DOI":"10.21767\/2248-9215.100004","article-title":"Antimicrobial activity of Trigonella foenum-graecum L. (fenugreek)","volume":"7","author":"Sharma","year":"","journal-title":"Eur J Exp Biol"},{"key":"ref39","doi-asserted-by":"publisher","first-page":"5392","DOI":"10.3892\/mmr.2020.11620","article-title":"In vitro anti-bacterial activity of diosgenin on Porphyromonas gingivalis and Prevotella intermedia","volume":"22","author":"Cong","year":"2020","journal-title":"Mol Med Rep"},{"key":"ref40","doi-asserted-by":"publisher","first-page":"122","DOI":"10.15406\/ppij.2015.02.00028","article-title":"Screening of antimicrobial activity of fenugreek seeds. Pharmacology","volume":"2","author":"Rehab","year":"2015","journal-title":"Int J"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"1450","DOI":"10.3390\/plants11111450","article-title":"Revisiting Trigonella foenum-graecum L.: pharmacology and therapeutic potentialities","volume":"11","author":"Visuvanathan","year":"2022","journal-title":"Plants"},{"key":"ref42","doi-asserted-by":"publisher","first-page":"207","DOI":"10.3390\/vetsci9050207","article-title":"Effect of graded levels of fenugreek (Trigonella foenum-graecum L.) seeds on the growth performance, hematological parameters, and intestinal histomorphology of broiler chickens","volume":"9","author":"Paneru","year":"2022","journal-title":"Vet Sci"},{"key":"ref43","doi-asserted-by":"publisher","first-page":"1245","DOI":"10.1038\/s41598-020-58005-7","article-title":"Fenugreek counters the effects of high fat diet on gut microbiota in mice: links to metabolic benefit","volume":"10","author":"Bruce-Keller","year":"2020","journal-title":"Sci Rep"},{"key":"ref44","doi-asserted-by":"publisher","first-page":"3654","DOI":"10.3390\/ijms23073654","article-title":"Cross-omics analysis of fenugreek supplementation reveals beneficial effects are caused by gut microbiome changes not mammalian host physiology","volume":"23","author":"Jones","year":"2022","journal-title":"Int J Mol Sci"},{"key":"ref45","doi-asserted-by":"publisher","first-page":"619","DOI":"10.29261\/pakvetj\/2024.207","article-title":"GC-MS analysis of chemical composition and determination of antimicrobial activity of Laurel leaf extracts prepared by different methods and solvents","volume":"3","author":"Ayse","year":"2024","journal-title":"Pak Vet J"},{"key":"ref46","doi-asserted-by":"publisher","first-page":"104130","DOI":"10.1016\/j.psj.2024.104130","article-title":"Immune modulation and cecal microbiome changes in broilers fed with fenugreek seeds and Bacillus-based probiotics","volume":"103","author":"Paneru","year":"2024","journal-title":"Poult Sci"},{"key":"ref47","doi-asserted-by":"publisher","first-page":"423","DOI":"10.1016\/j.sajb.2022.04.014","article-title":"Fenugreek (Trigonella foenum-graecum): nutraceutical values, phytochemical, ethnomedicinal and pharmacological overview","volume":"151","author":"Ruwali","year":"2022","journal-title":"S Afr J Bot"},{"key":"ref48","first-page":"1304","article-title":"Medicinal values of fenugreek \u2013 a review","volume":"4","author":"Mullaicharam","year":"2013","journal-title":"Res J Pharm, Biol Chem Sci"},{"key":"ref49","doi-asserted-by":"publisher","first-page":"592","DOI":"10.29261\/pakvetj\/2024.251","article-title":"A review on the use of phytochemicals for the control of zoonotic giardiasis","volume":"3","author":"Bader","year":"2024","journal-title":"Pak Vet J"},{"key":"ref50","doi-asserted-by":"publisher","first-page":"e000247","DOI":"10.1136\/vetreco-2017-000247","article-title":"Motile Salmonella serotypes causing high mortality in poultry farms in three South-Western states of Nigeria","volume":"4","author":"Mshelbwala","year":"2017","journal-title":"Vet Rec Open"},{"key":"ref51","doi-asserted-by":"publisher","first-page":"134","DOI":"10.1111\/j.1365-2052.2010.02090.x","article-title":"Genome-wide SNP analysis identifies major QTL for Salmonella colonization in the chicken","volume":"42","author":"Fife","year":"2011","journal-title":"Anim Genet"},{"key":"ref52","doi-asserted-by":"publisher","first-page":"259","DOI":"10.3390\/ph17020259","article-title":"Outlining the Phytoconstituents of Greek clover herb extract and assessment of its effect against foodborne infections caused by Salmonella typhimurium","volume":"17","author":"Alqahtani","year":"2024","journal-title":"Pharmaceuticals"},{"key":"ref53","doi-asserted-by":"publisher","first-page":"40053","DOI":"10.1007\/s11356-021-18358-6","article-title":"Comparative immunomodulatory efficacy of rosemary and fenugreek against Escherichia coli infection via suppression of inflammation and oxidative stress in broilers","volume":"29","author":"Farouk","year":"2022","journal-title":"Environ Sci Pollut Res"},{"key":"ref54","doi-asserted-by":"publisher","first-page":"289","DOI":"10.4038\/tar.v24i3.8014","article-title":"Effects of fenugreek (Trigonella foenum-graecum L.) seed powder on growth performance, visceral organ weight, serum cholesterol levels and the nitrogen retention of broiler or chicken","volume":"24","author":"Weerasingha","year":"2015","journal-title":"Trop Agric Res"},{"key":"ref55","doi-asserted-by":"publisher","first-page":"1298587","DOI":"10.3389\/fvets.2023.1298587","article-title":"Effect of fenugreek seeds and Bacillus-based direct-fed microbials on the growth performance, blood biochemicals, and intestinal histomorphology of broiler chickens","volume":"10","author":"Paneru","year":"2023","journal-title":"Front Vet Sci"},{"key":"ref56","doi-asserted-by":"publisher","first-page":"4","DOI":"10.29261\/pakvetj\/2024.299","article-title":"Use of polyphenols for the control of chicken meat borne zoonotic Campylobacter jejuni serotypes O1\/44, O2, and O4 complex","author":"Bekzhan","year":"2024","journal-title":"Pak Vet J"},{"key":"ref57","doi-asserted-by":"publisher","first-page":"957","DOI":"10.29261\/pakvetj\/2024.227","article-title":"Nanoparticles as potent allies in combating antibiotic resistance: a promising frontier in antimicrobial therapy","volume":"4","author":"Abdul","year":"2024","journal-title":"Pak Vet J"},{"key":"ref58","doi-asserted-by":"publisher","first-page":"311","DOI":"10.3390\/foods8080311","article-title":"Effect of a combination of fenugreek seeds, linseeds, garlic and copper sulfate on laying hens performances, egg physical and chemical qualities","volume":"8","author":"Omri","year":"2019","journal-title":"Foods"},{"key":"ref59","doi-asserted-by":"publisher","first-page":"47","DOI":"10.2141\/jpsa.0170011","article-title":"Egg production, egg quality, blood profiles, cecal microflora, and excreta noxious gas emission in laying hens fed with fenugreek (Trigonella foenum-graecum L.) seed extract","volume":"55","author":"Park","year":"2018","journal-title":"J Poult Sci"},{"key":"ref60","doi-asserted-by":"publisher","first-page":"1041","DOI":"10.3390\/ani10061041","article-title":"Characterisation and in vitro evaluation of fenugreek (Trigonella foenum-graecum) seed gum as a potential prebiotic in growing rabbit nutrition","volume":"10","author":"Zemzmi","year":"2020","journal-title":"Animals"}],"container-title":["Frontiers in Veterinary Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1570387\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,24]],"date-time":"2025-04-24T05:30:20Z","timestamp":1745472620000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2025.1570387\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,24]]},"references-count":60,"alternative-id":["10.3389\/fvets.2025.1570387"],"URL":"https:\/\/doi.org\/10.3389\/fvets.2025.1570387","relation":{},"ISSN":["2297-1769"],"issn-type":[{"value":"2297-1769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,24]]},"article-number":"1570387"}}