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Direct-fed microbials (DFM) can support gastrointestinal function and alleviate the potential negative effects of bacterial infections. In the present study, the capacity of a multispecies bacterial-based DFM containing Ligilactobacillus (formerly Lactobacillus) animalis 506, Propionibacterium freudenreichii 507, Bacillus licheniformis 809, and B. subtilis 597 to reduce S. Typhimurium ATCC14028 invasion was investigated using a co-incubation model with the HT29-MTX-E12 cell line (experiment 1). Next, a possible antagonistic effect of the DFM against S. Dublin ATCC 41286 was evaluated using an in vitro agar well diffusion method following a co-incubation of 48 h (experiment 2). At last, a series of experiments were performed to evaluate how different doses (6.25\u2005\u00d7\u2005106, 2.50\u2005\u00d7\u2005107, or 1.00\u2005\u00d7\u2005108 CFU\/well) of the DFM would support the integrity of intestinal epithelial cells challenged or not with S. Typhimurium ATCC14028 or hydrogen peroxide under a transepithelial electrical resistance (TEER) assay with Caco-2 cells (experiments 3 and 4). In experiment 1, BDP significantly (P\u2005&amp;lt;\u20050.001) reduced by 90.8% the invasion of S. Typhimurium into HT29-MTX-E12 cells, whereas viability of the potentially harmful bacteria was reduced by 21.0% (P\u2005&amp;lt;\u20050.0001). In experiment 2, the antagonistic properties of BDP towards S. Dublin were confirmed by the detection of a clear inhibition zone (size = 8.6 mm). Lastly, without challenge, the lowest dose of the DFM (6.25\u2005\u00d7\u2005106 CFU) provided the greatest support to the cells (treatment \u00d7 hour; P\u2005&amp;lt;\u20050.0001). However, when the cells were challenged with S. Typhimurium, all doses alleviated the loss of integrity caused by the pathogen (treatment \u00d7 hour; P\u2005&amp;lt;\u20050.0001). In cells challenged with hydrogen peroxide, the greater dose (1.00\u2005\u00d7\u2005108 CFU) supported the cells for a longer period of time (treatment \u00d7 hour; P\u2005&amp;lt;\u20050.0001). These in vitro findings set the stage for exploring the potential benefits of using a novel DFM as a promising tool and strategy to mitigate S. enterica infections in ruminants and improve animal health, food safety, and public health. Further, in vivo confirmation needs to be developed to validate these preliminary in vitro results.<\/jats:p>","DOI":"10.1093\/jas\/skae304","type":"journal-article","created":{"date-parts":[[2024,10,9]],"date-time":"2024-10-09T17:49:06Z","timestamp":1728496146000},"source":"Crossref","is-referenced-by-count":4,"title":["A multispecies bacterial-based direct-fed microbial alleviates <i>Salmonella<\/i> invasion and supports in vitro epithelial integrity"],"prefix":"10.1093","volume":"102","author":[{"given":"Raphaele","family":"Gresse","sequence":"first","affiliation":[{"name":"Novonesis, Planetary Health & Biosolutions , H\u00f8rsholm 2970 ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1819-4013","authenticated-orcid":false,"given":"Bruno I","family":"Cappellozza","sequence":"additional","affiliation":[{"name":"Novonesis, Planetary Health & Biosolutions , H\u00f8rsholm 2970 ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lena C","family":"Capern","sequence":"additional","affiliation":[{"name":"Novonesis, Planetary Health & Biosolutions , H\u00f8rsholm 2970 ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tine T M","family":"Knudsen","sequence":"additional","affiliation":[{"name":"Novonesis, Planetary Health & Biosolutions , H\u00f8rsholm 2970 ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Copani","sequence":"additional","affiliation":[{"name":"Novonesis, Planetary Health & Biosolutions , H\u00f8rsholm 2970 ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,10,9]]},"reference":[{"key":"2024111411245479300_CIT0001","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1111\/j.1574-6976.2010.00244.x","article-title":"Diversity and applications of Bacillus bacteriocins","volume":"35","author":"Abriouel","year":"2011","journal-title":"FEMS Microbiol. 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