{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T17:40:07Z","timestamp":1789839607209,"version":"4.0.1"},"reference-count":47,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,2,10]],"date-time":"2025-02-10T00:00:00Z","timestamp":1739145600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32072748"],"award-info":[{"award-number":["32072748"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The modern livestock industry faces significant challenges due to the extension of production cycles and a growing shortage of feed resources. Low-protein (LP) diets offer potential environmental advantages whereas damage intestinal integrity and increase pro-inflammatory cytokines. Bile acids (BAs), metabolic products of cholesterol in the liver, possess strong antioxidative and immune properties. This study evaluated the effects of dietary porcine BA supplementation at different protein levels on the antioxidant status, immune function, and gut microbiota of late-laying hens. A 2 \u00d7 2 factorial design was employed with 198 Hy-Line Brown laying hens (62 weeks old), randomly divided into one of four treatment groups with 8 replicates for each treatment. Diets included normal (16.42%) or low-protein (15.35%) levels, with porcine BAs supplemented at 0 or 120 mg\/kg (62\u201369 weeks) and 0 or 200 mg\/kg (70\u201375 weeks) over 14 weeks. Dietary protein levels and bile acids had no effect on organ development in laying hens. Low-protein diets increased serum anti-inflammatory cytokines IL-4 and IL-10 but elevated pro-inflammatory cytokines IL-1\u03b2 (p &lt; 0.05), corroborated by higher IL-10 gene expression in the ileum mucosa (p &lt; 0.05). The supplementation of bile acids improved immune function by increasing serum IL-4 and TGF-\u03b2 while decreasing IL-6 levels (p &lt; 0.05). Moreover, bile acids treatments upregulated IL-10, TGF-\u03b2 and ZO-1 gene expression in the ileum mucosa (p &lt; 0.05) and alleviated oxidative stress by enhancing serum GSH-Px activity (p &lt; 0.05). Additionally, both low-protein diets and bile acids enriched beneficial cecal bacteria, including Hungatella and Blautia, primarily linked to immune function. These findings demonstrate that porcine bile acids administration improve antioxidant capacity, immune function and gut microbiota dysbiosis. These beneficial effects were associated with Hungatella and Blautia levels, which may be responsible for the porcine bile acids efficacy in poultry nutrition.<\/jats:p>","DOI":"10.3390\/ani15040500","type":"journal-article","created":{"date-parts":[[2025,2,11]],"date-time":"2025-02-11T11:54:06Z","timestamp":1739274846000},"page":"500","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Porcine Bile Acids Improve Antioxidant Status and Immune Function by Increasing Hungatella Abundance with Different Protein Level Diets in Late-Laying Hens"],"prefix":"10.3390","volume":"15","author":[{"given":"Ronghui","family":"Xing","sequence":"first","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengfei","family":"Du","sequence":"additional","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyang","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zongze","family":"Fan","sequence":"additional","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Longfei","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanqun","family":"Huang","sequence":"additional","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuemeng","family":"Si","sequence":"additional","affiliation":[{"name":"Institute of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1234","DOI":"10.14202\/vetworld.2020.1234-1244","article-title":"Supplementation of a low-protein diet with tryptophan, threonine, and valine and its impact on growth performance, blood biochemical constituents, immune parameters, and carcass traits in broiler chickens","volume":"13","author":"Reham","year":"2020","journal-title":"Vet. World"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"101465","DOI":"10.1016\/j.psj.2021.101465","article-title":"Effect of dietary serine supplementation on performance, egg quality, serum indices, and ileal mucosal immunity in laying hens fed a low crude protein diet","volume":"100","author":"Zhou","year":"2021","journal-title":"Poult. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"102798","DOI":"10.1016\/j.psj.2023.102798","article-title":"Effects of dietary protein levels on performance, nitrogen excretion, and odor emission of growing pullets and laying hens","volume":"102","author":"Heo","year":"2023","journal-title":"Poult. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1016\/j.tips.2017.04.005","article-title":"Targeting free radicals in oxidative stress-related human diseases","volume":"38","author":"Poprac","year":"2017","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Li, J., Zhou, D., Li, H., Luo, Q., Wang, X., Qin, J., Xu, Y., Lu, Q., and Tian, X. (2022). Effect of purple corn extract on performance, antioxidant activity, egg quality, egg amino acid, and fatty acid profiles of laying hen. Front. Vet. Sci., 9.","DOI":"10.3389\/fvets.2022.1083842"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1152\/physrev.00026.2013","article-title":"Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release","volume":"94","author":"Zorov","year":"2014","journal-title":"Physiol. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.cmet.2016.05.005","article-title":"Intestinal crosstalk between bile acids and microbiota and its impact on host metabolism","volume":"24","author":"Sayin","year":"2016","journal-title":"Cell Metab."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1152\/physrev.00010.2008","article-title":"Role of bile acids and bile acid receptors in metabolic regulation","volume":"89","author":"Lefebvre","year":"2009","journal-title":"Physiol. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e0080521","DOI":"10.1128\/msystems.00805-21","article-title":"Dominant bacterial phyla from the human gut show widespread ability to transform and conjugate bile acids","volume":"6","author":"Lucas","year":"2021","journal-title":"mSystems"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Zhu, F., Zheng, S., Zhao, M., Shi, F., Zheng, L., and Wang, H. (2023). The regulatory role of bile acid microbiota in the progression of liver cirrhosis. Front. Pharmacol., 14.","DOI":"10.3389\/fphar.2023.1214685"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Li, W., Zhang, Y., Yang, J., Xu, H., Ye, R., Wu, J., Cao, M., Zhao, C., Yang, B., and Liu, C. (2024). Effect of bile acids supplementation in fatty liver hemorrhagic syndrome, production performance, physiological and quality characteristics of laying hen eggs. Animals, 14.","DOI":"10.3390\/ani14131910"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Li, Z., Yuan, H., Chu, H., and Yang, L. (2023). The crosstalk between gut microbiota and bile acids promotes the development of non-alcoholic fatty liver disease. Microorganisms, 11.","DOI":"10.3390\/microorganisms11082059"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1038\/s41586-019-1785-z","article-title":"Bile acid metabolites control T(H)17 and T(reg) cell differentiation","volume":"576","author":"Hang","year":"2019","journal-title":"Nature"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1038\/s41586-020-2193-0","article-title":"Bacterial metabolism of bile acids promotes generation of peripheral regulatory T cells","volume":"581","author":"Campbell","year":"2020","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Gr\u00fcner, N., and Mattner, J. (2021). Bile acids and microbiota: Multifaceted and versatile regulators of the liver\u2013gut axis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22031397"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1016\/j.intimp.2012.11.017","article-title":"Dose-dependent antiinflammatory effect of ursodeoxycholic acid in experimental colitis","volume":"15","author":"Requena","year":"2013","journal-title":"Int. Immunopharmacol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Sannasiddappa, T.H., Lund, P.A., and Clarke, S.R. (2017). In vitro antibacterial activity of unconjugated and conjugated bile salts on staphylococcus aureus. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.01581"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"172393","DOI":"10.1016\/j.ejphar.2019.05.022","article-title":"Obeticholic acid protects against diabetic cardiomyopathy by activation of FXR\/Nrf2 signaling in db\/db mice","volume":"858","author":"Wu","year":"2019","journal-title":"Eur. J. Pharmacol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Liu, Y., Azad, M.A.K., Kong, X., Zhu, Q., and Yu, Z. (2022). Dietary bile acids supplementation modulates immune response, antioxidant capacity, glucose, and lipid metabolism in normal and intrauterine growth retardation piglets. Front. Nutr., 9.","DOI":"10.3389\/fnut.2022.991812"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Yin, C., Xia, B., Tang, S., Cao, A., Liu, L., Zhong, R., Chen, L., and Zhang, H. (2021). The effect of exogenous bile acids on antioxidant status and gut microbiota in heat-stressed broiler chickens. Front. Nutr., 8.","DOI":"10.3389\/fnut.2021.747136"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"184","DOI":"10.1016\/j.chroma.2014.10.064","article-title":"A liquid chromatography-tandem mass spectrometry-based method for the simultaneous determination of hydroxy sterols and bile acids","volume":"1371","author":"John","year":"2014","journal-title":"J. Chromatogr. A"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Yang, B., Huang, S., Li, S., Feng, Z., Zhao, G., and Ma, Q. (2022). Safety Evaluation of Porcine Bile Acids in Laying Hens: Effects on laying performance, egg quality, blood parameters, organ indexes, and intestinal development. Front. Vet. Sci., 9.","DOI":"10.3389\/fvets.2022.895831"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1016\/j.aninu.2022.08.003","article-title":"Dietary supplementation of porcine bile acids improves laying performance, serum lipid metabolism and cecal microbiota in late-phase laying hens","volume":"11","author":"Yang","year":"2022","journal-title":"Anim. Nutr."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"skad334","DOI":"10.1093\/jas\/skad334","article-title":"Effect of exogenous bile salts supplementation on the performance and hepatic lipid metabolism of aged laying hens","volume":"101","author":"Sun","year":"2023","journal-title":"J. Anim. Sci."},{"key":"ref_25","unstructured":"Hy-Line International (2018). Hy-Line Brown Management Guide, Hy-Line International."},{"key":"ref_26","unstructured":"AOAC (2005). Official Methods of Analysis of AOAC International, AOAC International. [18th ed.]. Available online: https:\/\/www.aoac.org\/."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1111\/j.1439-0396.2008.00898.x","article-title":"Long-term feeding effects of dietary protein levels on egg production, immunocompetence and plasma amino acids of laying hens in subtropical condition","volume":"94","author":"Poosuwan","year":"2010","journal-title":"J. Anim. Physiol. Anim. Nutr."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Kim, D.H., Lee, Y.K., Lee, S.D., and Lee, K.W. (2022). Impact of relative humidity on the laying performance, egg quality, and physiological stress responses of laying hens exposed to high ambient temperature. J. Therm. Biol., 103.","DOI":"10.1016\/j.jtherbio.2021.103167"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e2004433","DOI":"10.1002\/advs.202004433","article-title":"Cytokines: From clinical significance to quantification","volume":"8","author":"Liu","year":"2021","journal-title":"Adv. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"718","DOI":"10.4049\/jimmunol.1700183","article-title":"The bile acid receptor GPBAR1 regulates the M1\/M2 phenotype of intestinal macrophages and activation of GPBAR1 rescues mice from murine colitis","volume":"199","author":"Biagioli","year":"2017","journal-title":"J. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1038\/s41385-019-0162-4","article-title":"Emerging roles of bile acids in mucosal immunity and inflammation","volume":"12","author":"Chen","year":"2019","journal-title":"Mucosal Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"81","DOI":"10.31083\/j.rcm2403081","article-title":"Role of the stress- and inflammation-induced cytokine GDF-15 in cardiovascular diseases: From basic research to clinical relevance","volume":"24","author":"Schwarz","year":"2023","journal-title":"Rev. Cardiovasc. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1016\/j.clinbiochem.2010.07.022","article-title":"Malondialdehyde (MDA) and protein carbonyl (PCO) levels as biomarkers of oxidative stress in subjects with familial hypercholesterolemia","volume":"43","author":"Pirinccioglu","year":"2010","journal-title":"Clin. Biochem."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Attia, Y.A., Al-Harthi, M.A., and Abo El-Maaty, H.M. (2020). The effects of different Oil sources on performance, digestive enzymes, carcass traits, biochemical, immunological, antioxidant, and morphometric responses of broiler chicks. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.00181"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/j.bpg.2007.11.002","article-title":"The mucosal immune system at the gastrointestinal barrier","volume":"22","author":"Schenk","year":"2008","journal-title":"Best Pract. Res. Clin. Gastroenterol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"6549","DOI":"10.1016\/j.psj.2020.09.038","article-title":"Effects of glucose oxidase on growth performance, immune function and intestinal barrier of ducks infected with Escherichia coli O88","volume":"99","author":"Liu","year":"2020","journal-title":"Poult. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1111\/bph.14095","article-title":"Tauroursodeoxycholic acid inhibits intestinal inflammation and barrier disruption in mice with non-alcoholic fatty liver disease","volume":"175","author":"Wang","year":"2018","journal-title":"Br. J. Pharmacol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e2200649","DOI":"10.1002\/mnfr.202200649","article-title":"Bile acid derivatives effectively prevented high-fat diet-induced colonic barrier dysfunction","volume":"67","author":"Ma","year":"2023","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"277","DOI":"10.4110\/in.2014.14.6.277","article-title":"Gut microbiota-derived short-chain fatty acids, T cells, and inflammation","volume":"14","author":"Kim","year":"2014","journal-title":"Immune Netw."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e51","DOI":"10.1097\/MD.0000000000000051","article-title":"Characteristics of fecal and mucosa-associated microbiota in Chinese patients with inflammatory bowel disease","volume":"93","author":"Chen","year":"2014","journal-title":"Medicine"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Rawat, P.S., Li, Y., Zhang, W., Meng, X., and Liu, W. (2022). Hungatella hathewayi, an efficient glycosaminoglycan-degrading firmicutes from human gut and its chondroitin ABC exolyase with high activity and broad substrate specificity. Appl. Environ. Microbiol., 88.","DOI":"10.1128\/aem.01546-22"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2068367","DOI":"10.1080\/19490976.2022.2068367","article-title":"Utilization of glycosaminoglycans by the human gut microbiota: Participating bacteria and their enzymatic machineries","volume":"14","author":"Rawat","year":"2022","journal-title":"Gut Microbes."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1007\/s00394-021-02649-2","article-title":"Dietary inflammatory index and its relationship with gut microbiota in individuals with intestinal constipation: A cross-sectional study","volume":"61","author":"Costa","year":"2022","journal-title":"Eur. J. Nutr."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Cani, P.D., and de Vos, W.M. (2017). Next-generation beneficial microbes: The case of akkermansia muciniphila. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.01765"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1016\/j.celrep.2017.03.080","article-title":"NLRP6 Protects Il10(-\/-) mice from colitis by limiting colonization of akkermansia muciniphila","volume":"19","author":"Seregin","year":"2017","journal-title":"Cell Rep."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1111\/imm.12760","article-title":"The immune response to prevotella bacteria in chronic inflammatory disease","volume":"151","author":"Larsen","year":"2017","journal-title":"Immunology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2281014","DOI":"10.1080\/19490976.2023.2281014","article-title":"Gut-microbiota prompt activation of natural killer cell on alcoholic liver disease","volume":"15","author":"Eom","year":"2023","journal-title":"Gut Microbes."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/4\/500\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:30:32Z","timestamp":1760027432000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/15\/4\/500"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,10]]},"references-count":47,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,2]]}},"alternative-id":["ani15040500"],"URL":"https:\/\/doi.org\/10.3390\/ani15040500","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,10]]}}}