{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:37:27Z","timestamp":1759970247288,"version":"build-2065373602"},"reference-count":39,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2025,1,9]],"date-time":"2025-01-09T00:00:00Z","timestamp":1736380800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Guizhou Provincial Basic Research Program (Natural Science)","award":["QKHJC-ZK-2022-129","QKHJC-ZK-2023-109","32202854","32360899","QJH-KY-2022-151"],"award-info":[{"award-number":["QKHJC-ZK-2022-129","QKHJC-ZK-2023-109","32202854","32360899","QJH-KY-2022-151"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["QKHJC-ZK-2022-129","QKHJC-ZK-2023-109","32202854","32360899","QJH-KY-2022-151"],"award-info":[{"award-number":["QKHJC-ZK-2022-129","QKHJC-ZK-2023-109","32202854","32360899","QJH-KY-2022-151"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Guizhou Province General Colleges and Universities Youth Science and Technology Talent Growth Project","award":["QKHJC-ZK-2022-129","QKHJC-ZK-2023-109","32202854","32360899","QJH-KY-2022-151"],"award-info":[{"award-number":["QKHJC-ZK-2022-129","QKHJC-ZK-2023-109","32202854","32360899","QJH-KY-2022-151"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Isoflavone components extracted from red clover have anti-inflammatory, antioxidant and immune boosting effects. We hypothesize that red clover isoflavones (RCIs) achieve health-promoting effects via altering the gut microbiota. A total of 48 mice (20 \u00b1 2 g) were randomly divided into a control group, low-dose group (0.05% RCIs in feed), middle-dose group (0.1% RCIs in feed), and high-dose group (0.2% RCIs in feed) with 12 mice per group. The feeding period was 20 d. The results showed that RCIs can increase the daily gain and decrease the ratio of feed to gain in mice. The organ indexes and blood biochemical indexes of the mice in each RCI group were in the normal range, indicating that RCIs do not damage liver or kidney function. RCI supplementation increased serum immunity and altered the microbial community structure in the cecum of the mice. RCIs can increase the diversity of beneficial bacteria such as Bacteroidaceae, Muribaculaceae, and Akkermansiaceae, and reduced the pathogenic Staphylococcaceae. Therefore, supplementing the diet with RCIs results in improved growth performance and notable alterations in the cecal microbiota in mice, and has potential applications as a feed additive to improve livestock production.<\/jats:p>","DOI":"10.3390\/ani15020150","type":"journal-article","created":{"date-parts":[[2025,1,9]],"date-time":"2025-01-09T05:17:08Z","timestamp":1736399828000},"page":"150","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Effects of Red Clover Isoflavones on Growth Performance, Immune Function, and Cecal Microflora of Mice"],"prefix":"10.3390","volume":"15","author":[{"given":"Rongrong","family":"Guo","sequence":"first","affiliation":[{"name":"Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8671-3161","authenticated-orcid":false,"given":"Xuqin","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaodie","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-1058-6005","authenticated-orcid":false,"given":"Cheng","family":"Zeng","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8434-0127","authenticated-orcid":false,"given":"Ying","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunjie","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Pulmonary and Inflammation, Frontiers Science Center for Disease-Related Molecular Network, Sichuan University, Chengdu 610000, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jian","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Deyuan","family":"Ou","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, College of Animal Science, Guizhou University, Guiyang 550025, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,1,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Chen, G., Li, Z., Liu, S., Tang, T., Chen, Q., Yan, Z., Peng, J., Yang, Z., Zhang, G., and Liu, Y. (2023). Fermented Chinese Herbal Medicine Promoted Growth Performance, Intestinal Health, and Regulated Bacterial Microbiota of Weaned Piglets. Animals, 13.","DOI":"10.3390\/ani13030476"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"110868","DOI":"10.1016\/j.foodres.2021.110868","article-title":"The bioavailability of soy isoflavones in vitro and their effects on gut microbiota in the simulator of the human intestinal microbial ecosystem","volume":"152","author":"Chen","year":"2022","journal-title":"Food Res. Int."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"106676","DOI":"10.1016\/j.phrs.2023.106676","article-title":"Dietary genistein increases microbiota-derived short chain fatty acid levels, modulates homeostasis of the aging gut, and extends healthspan and lifespan","volume":"188","author":"Hou","year":"2023","journal-title":"Pharmacol. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"10879","DOI":"10.1021\/acs.jafc.3c09444","article-title":"Puerarin Delays Mammary Gland Aging by Regulating Gut Microbiota and Inhibiting the p38MAPK Signaling Pathway","volume":"72","author":"Liu","year":"2024","journal-title":"J. Agric. Food Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1016\/j.aninu.2023.12.005","article-title":"Effect of red clover isoflavones on hormone, immune, inflammatory, and plasma biochemistry in lactating dairy cows","volume":"16","author":"Zhang","year":"2024","journal-title":"Anim. Nutr."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Luo, Z., Xu, Y., Qiu, L., Lv, S., Zeng, C., Tan, A., Ou, D., Song, X., and Yang, J. (2023). Optimization of ultrasound-assisted extraction based on response surface methodology using HPLC-DAD for the analysis of red clover (Trifolium pretense L.) isoflavones and its anti-inflammatory activities on LPS-induced 3D4\/2 cell. Front. Vet. Sci., 10.","DOI":"10.3389\/fvets.2023.1279178"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Breikaa, R.M., Algandaby, M.M., El-Demerdash, E., and Abdel-Naim, A.B. (2013). Multimechanistic antifibrotic effect of biochanin a in rats: Implications of proinflammatory and profibrogenic mediators. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069276"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"138764","DOI":"10.1016\/j.foodchem.2024.138764","article-title":"Enrichment of milk antioxidant activity by dietary supplementation of red clover isoflavone in cows and its improvement on mice intestinal health","volume":"446","author":"Zhang","year":"2024","journal-title":"Food Chem."},{"key":"ref_9","first-page":"1175","article-title":"Application of red clover isoflavone extract as an adjuvant in mice","volume":"19","author":"Chen","year":"2020","journal-title":"Exp. Ther. Med."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Feng, J., Sun, D., Wang, L., Li, X., Guan, J., Wei, L., Yue, D., Wang, X., Zhao, Y., and Yang, H. (2021). Biochanin A as an alpha-hemolysin inhibitor for combating methicillin-resistant Staphylococcus aureus infection. World J. Microbiol. Biotechnol., 38.","DOI":"10.1007\/s11274-021-03182-4"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"11987","DOI":"10.1039\/D1FO00007A","article-title":"Neuroprotective mechanisms of red clover and soy isoflavones in Parkinson\u2019s disease models","volume":"12","author":"Ambaw","year":"2021","journal-title":"Food Funct."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.aninu.2020.11.004","article-title":"The impact of probiotics on gut health via alternation of immune status of monogastric animals","volume":"7","author":"Ding","year":"2021","journal-title":"Anim. Nutr."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Sun, W., Chen, Z., Huang, Z., Wan, A., Zhou, M., and Gao, J. (2023). Effects of dietary traditional Chinese medicine residues on growth performance, intestinal health and gut microbiota compositions in weaned piglets. Front. Cell Infect. Microbiol., 13.","DOI":"10.3389\/fcimb.2023.1283789"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1111\/jam.12972","article-title":"Effect of Bacillus subtilis CGMCC 1.1086 on the growth performance and intestinal microbiota of broilers","volume":"120","author":"Li","year":"2016","journal-title":"J. Appl. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1111\/jpn.13929","article-title":"Dietary supplementation of water extract of Eucommia ulmoides bark improved caecal microbiota and parameters of health in white-feathered broilers","volume":"108","author":"Lu","year":"2024","journal-title":"J. Anim. Physiol. Anim. Nutr."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Xiao, C., Li, K., Teng, C., Wei, Z., Li, J., Zhang, S., Liu, L., Lv, H., and Zhong, R. (2023). Dietary Qi-Weng-Huangbo powder enhances growth performance, diarrhoea and immune function of weaned piglets by modulating gut health and microbial profiles. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1342852"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1111\/jpn.13247","article-title":"Effects of dietary supplementation of Lycium barbarum polysaccharides on growth performance, immune status, antioxidant capacity and selected microbial populations of weaned piglets","volume":"104","author":"Chen","year":"2020","journal-title":"J. Anim. Physiol. Anim. Nutr."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Ragland, S.A., and Criss, A.K. (2017). From bacterial killing to immune modulation: Recent insights into the functions of lysozyme. PLoS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006512"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"895","DOI":"10.1016\/S2221-1691(12)60249-4","article-title":"Effect of Commiphora mukul gum resin on hepatic marker enzymes, lipid peroxidation and antioxidants status in pancreas and heart of streptozotocin induced diabetic rats","volume":"2","author":"Ramesh","year":"2012","journal-title":"Asian Pac. J. Trop. Biomed."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1194","DOI":"10.1016\/j.fct.2012.02.008","article-title":"Quercetin prevents ethanol-induced dyslipidemia and mitochondrial oxidative damage","volume":"50","author":"Tang","year":"2012","journal-title":"Food Chem. Toxicol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1186\/s12934-019-1161-6","article-title":"Probiotics isolated from yaks improves the growth performance, antioxidant activity, and cytokines related to immunity and inflammation in mice","volume":"18","author":"Li","year":"2019","journal-title":"Microb. Cell Fact."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4508","DOI":"10.4049\/jimmunol.175.7.4508","article-title":"Two divergent isotypes of the fourth complement component from a bony fish, the common carp (Cyprinus carpio)","volume":"175","author":"Mutsuro","year":"2005","journal-title":"J. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1111\/asj.12444","article-title":"Effects of dietary lysozyme levels on growth performance, intestinal morphology, non-specific immunity and mRNA expression in weanling piglets","volume":"87","author":"Long","year":"2016","journal-title":"Anim. Sci. J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"269","DOI":"10.2478\/jvetres-2018-0038","article-title":"Saponin-based Mycoplasma Bovis Vaccine Containing Lysozyme Dimer Adjuvant Stimulates Acute Phase Response in Calves","volume":"62","author":"Dudek","year":"2018","journal-title":"J. Vet. Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.cyto.2014.02.001","article-title":"Human monocytes produce interferon-gamma upon stimulation with LPS","volume":"67","author":"Kraaij","year":"2014","journal-title":"Cytokine"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Wei, X., Bottoms, K.A., Stein, H.H., Blavi, L., Bradley, C.L., Bergstrom, J., Knapp, J., Story, R., Maxwell, C., and Tsai, T. (2021). Dietary Organic Acids Modulate Gut Microbiota and Improve Growth Performance of Nursery Pigs. Microorganisms, 9.","DOI":"10.3390\/microorganisms9010110"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Zhang, H., Shao, M., Huang, H., Wang, S., Ma, L., Wang, H., Hu, L., Wei, K., and Zhu, R. (2018). The Dynamic Distribution of Small-Tail Han Sheep Microbiota across Different Intestinal Segments. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.00032"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.aninu.2021.05.001","article-title":"Intestinal microbiota and its interaction to intestinal health in nursery pigs","volume":"8","author":"Duarte","year":"2022","journal-title":"Anim. Nutr."},{"key":"ref_29","first-page":"e2866","article-title":"Sustaining exposure to high concentrations of bifidobacteria inhibits gene expression of Mouse\u2019s mucosal immunity","volume":"5","author":"El","year":"2019","journal-title":"Heliyon"},{"key":"ref_30","first-page":"220","article-title":"The Immunostimulatory Effect of Lactic Acid Bacteria in a Rat Model","volume":"13","author":"Karamese","year":"2016","journal-title":"Iran. J. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.cell.2005.05.007","article-title":"An immunomodulatory molecule of symbiotic bacteria directs maturation of the host immune system","volume":"122","author":"Mazmanian","year":"2005","journal-title":"Cell"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Zhang, J., Shu, Z., Lv, S., Zhou, Q., Huang, Y., Peng, Y., Zheng, J., Zhou, Y., Hu, C., and Lan, S. (2023). Fermented Chinese Herbs Improve the Growth and Immunity of Growing Pigs through Regulating Colon Microbiota and Metabolites. Animals, 13.","DOI":"10.3390\/ani13243867"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"100279","DOI":"10.1016\/j.vas.2022.100279","article-title":"Determining the association between gut microbiota and its metabolites with higher intestinal Immunoglobulin A response","volume":"19","author":"Rajput","year":"2023","journal-title":"Vet. Anim. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1696","DOI":"10.1039\/C8FO02335B","article-title":"Dietary lysozyme supplementation contributes to enhanced intestinal functions and gut microflora of piglets","volume":"10","author":"Xiong","year":"2019","journal-title":"Food Funct."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Zhu, Y., Chen, B., Zhang, X., Akbar, M.T., Wu, T., Zhang, Y., Zhi, L., and Shen, Q. (2024). Exploration of the Muribaculaceae Family in the Gut Microbiota: Diversity, Metabolism, and Function. Nutrients, 16.","DOI":"10.3390\/nu16162660"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1080\/19490976.2015.1134082","article-title":"Formation of short chain fatty acids by the gut microbiota and their impact on human metabolism","volume":"7","author":"Morrison","year":"2016","journal-title":"Gut Microbes"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"937","DOI":"10.1007\/s10482-023-01857-6","article-title":"Phylogenomic and molecular markers based studies on Staphylococcaceae and Gemella species. Proposals for an emended family Staphylococcaceae and three new families (Abyssicoccaceae fam. nov., Salinicoccaceae fam. nov. and Gemellaceae fam. nov.) harboring four new genera, Lacicoccus gen. nov., Macrococcoides gen. nov., Gemelliphila gen. nov., and Phocicoccus gen. nov","volume":"116","author":"Bello","year":"2023","journal-title":"Antonie Van. Leeuwenhoek"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Ouyang, J., Lin, J., Isnard, S., Fombuena, B., Peng, X., Marette, A., Routy, B., Messaoudene, M., Chen, Y., and Routy, J.P. (2020). The Bacterium Akkermansia muciniphila: A Sentinel for Gut Permeability and Its Relevance to HIV-Related Inflammation. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.00645"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Bian, X., Wu, W., Yang, L., Lv, L., Wang, Q., Li, Y., Ye, J., Fang, D., Wu, J., and Jiang, X. (2019). Administration of Akkermansia muciniphila Ameliorates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.02259"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/2\/150\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,8]],"date-time":"2025-10-08T10:25:37Z","timestamp":1759919137000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/2\/150"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,1,9]]},"references-count":39,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["ani15020150"],"URL":"https:\/\/doi.org\/10.3390\/ani15020150","relation":{},"ISSN":["2076-2615"],"issn-type":[{"type":"electronic","value":"2076-2615"}],"subject":[],"published":{"date-parts":[[2025,1,9]]}}}