{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:42:15Z","timestamp":1760060535427,"version":"build-2065373602"},"reference-count":59,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T00:00:00Z","timestamp":1756425600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry of University and Research under National Recovery and Resilience Plan (NRRP)","award":["D93C22000890001","PE00000003"],"award-info":[{"award-number":["D93C22000890001","PE00000003"]}]},{"name":"ON Foods\u2014Research and innovation network on food and nutrition Sustainability, Safety and Security\u2014Working ON Foods","award":["D93C22000890001","PE00000003"],"award-info":[{"award-number":["D93C22000890001","PE00000003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Nowadays, the consumption of \u201cfree from\u201d foods by non-specific consumers is increasing, partly due to a misperception of labels that make them seem healthier. These foods are formulated for consumers with allergies or diseases that limit their diet, and it is not known if there are more benefits than risks for healthy consumers. For example, there is no work investigating the interaction between lactose-free milk and the colonic microbiome of healthy individuals. To focus on the potential modulation of gut microbiota of healthy subjects by lactose-free milk, we performed an in vitro simulation of digestion and fermentation, integrating microbiomics and metabolomics approaches to study changes in gut microbiota populations and metabolite production. Results indicated that lactose-free and lactose-containing milk differently modulated colonic microbiota based on several microbiological indicators, including the reduction in Bifidobacteriaceae (approximately more than two times) and Lactobacillales and the reduction in the beneficial production of microbial compounds (approximately six times less acetic acid and two times less butanoic acid). Such features suggest that lactose-free milk increases the risk of dysbiosis in healthy subjects. Our work identifies the drivers of this dysbiosis among hundreds of molecules and microbes of the gut microbiota, assigning specific names and ecological niches for the first time. It employs an in vitro model, which represents a new standard for sustainable research and improves translatability. Our findings support the European Society for Clinical Nutrition and Metabolism (ESPEN) guidelines, which do not recommend the routine consumption of lactose-free diets in the absence of diagnosed intolerance.<\/jats:p>","DOI":"10.3390\/microorganisms13092021","type":"journal-article","created":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T16:18:15Z","timestamp":1756484295000},"page":"2021","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Are \u201cFree From\u201d Foods Risk-Free? Lactose-Free Milk Fermentation Modulates Normal Colon in a Gut Microbiota in Vitro Model"],"prefix":"10.3390","volume":"13","author":[{"given":"Flavia","family":"Casciano","sequence":"first","affiliation":[{"name":"DiSTAL\u2014Department of Agricultural and Food Sciences, <i>Alma Mater Studiorum<\/i>\u2014University of Bologna, P.za G. Goidanich, 60, 47521 Cesena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7875-6321","authenticated-orcid":false,"given":"Lorenzo","family":"Nissen","sequence":"additional","affiliation":[{"name":"DiSTAL\u2014Department of Agricultural and Food Sciences, <i>Alma Mater Studiorum<\/i>\u2014University of Bologna, P.za G. Goidanich, 60, 47521 Cesena, Italy"},{"name":"CIRI\u2014Interdepartmental Centre of Agri-Food Industrial Research, <i>Alma Mater Studiorum<\/i>\u2014University of Bologna, P.za G. Goidanich, 60, 47521 Cesena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4579-1662","authenticated-orcid":false,"given":"Alessandra","family":"Bordoni","sequence":"additional","affiliation":[{"name":"DiSTAL\u2014Department of Agricultural and Food Sciences, <i>Alma Mater Studiorum<\/i>\u2014University of Bologna, P.za G. Goidanich, 60, 47521 Cesena, Italy"},{"name":"CIRI\u2014Interdepartmental Centre of Agri-Food Industrial Research, <i>Alma Mater Studiorum<\/i>\u2014University of Bologna, P.za G. Goidanich, 60, 47521 Cesena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8847-6251","authenticated-orcid":false,"given":"Andrea","family":"Gianotti","sequence":"additional","affiliation":[{"name":"DiSTAL\u2014Department of Agricultural and Food Sciences, <i>Alma Mater Studiorum<\/i>\u2014University of Bologna, P.za G. Goidanich, 60, 47521 Cesena, Italy"},{"name":"CIRI\u2014Interdepartmental Centre of Agri-Food Industrial Research, <i>Alma Mater Studiorum<\/i>\u2014University of Bologna, P.za G. Goidanich, 60, 47521 Cesena, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1093\/ajcn\/nqz104","article-title":"Lactose digestion in humans: Intestinal lactase appears to be constitutive whereas the colonic microbiome is adaptable","volume":"110","year":"2019","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"157","DOI":"10.3184\/003685005783238408","article-title":"The molecular basis of lactose intolerance","volume":"88","author":"Campbell","year":"2005","journal-title":"Sci. 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