{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,13]],"date-time":"2026-09-13T05:04:44Z","timestamp":1789275884011,"version":"build-2803163510"},"reference-count":47,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:00:00Z","timestamp":1789084800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Major Science and Technology Special Program of Henan Province","award":["251100110400"],"award-info":[{"award-number":["251100110400"]}]},{"name":"National Key Research And Development Program Of China","award":["2021YFD1300300"],"award-info":[{"award-number":["2021YFD1300300"]}]},{"name":"Henan International Cooperation and Exchange Program","award":["252102520031"],"award-info":[{"award-number":["252102520031"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Enterotoxigenic Escherichia coli (ETEC) infection severely impairs post-weaning piglet health, causing intestinal dysfunction, systemic inflammation, and significant economic losses in the swine industry. This study aims to evaluate a synthesized palmatine-intercalated montmorillonite composite (MP) for improving growth performance, immune response, and intestinal health in ETEC-challenged piglets. Following structural characterization, an in vivo trial was conducted using 30 crossbred piglets (Yorkshire \u00d7 Landrace \u00d7 Duroc; 15.12 \u00b1 2.95 kg BW) challenged with ETEC K88 and supplemented with 0.02%, 0.5%, and 1.5% MP. Structural analysis confirmed successful intercalation of palmatine into montmorillonite, with enhanced antibacterial and anti-inflammatory properties. In vivo, dietary inclusion of the 1.5% MP significantly alleviated ETEC-induced growth depression, as evidenced by increased average daily gain and improved apparent total tract digestibility of crude protein. These effects were associated with improved jejunal morphology and upregulation of PEPT1, B0AT1, and SGLT1. Moreover, MP supplementation attenuated systemic inflammation and hepatic stress, as indicated by reduced serum AST, TNF-\u03b1, and IL-1\u03b2 levels. Microbiota analysis showed that MP modulated ETEC-associated alterations in the cecal microbial community. In conclusion, the MP represents a promising nutritional approach to improve intestinal resilience, nutrient utilization, and growth performance of ETEC-challenged piglets.<\/jats:p>","DOI":"10.3390\/ani16182868","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T14:39:24Z","timestamp":1789137564000},"page":"2868","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Palmatine-Intercalated Montmorillonite Improves Intestinal Health and Attenuates Systemic Inflammation in ETEC-Challenged Piglets"],"prefix":"10.3390","volume":"16","author":[{"given":"Liping","family":"Gan","sequence":"first","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Yifeng","family":"Zhao","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Shiya","family":"Li","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Yingying","family":"Sheng","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"}]},{"given":"Dongtian","family":"Zhao","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"given":"Yuan","family":"Yuan","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"investigation","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"}]},{"given":"Hanzhen","family":"Qiao","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"supervision","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7737-8448","authenticated-orcid":false,"given":"Peng","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"given":"Erzhen","family":"Duan","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"given":"Tengpeng","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1450-1784","authenticated-orcid":false,"given":"Jinrong","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Bioengineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.aninu.2025.10.010","article-title":"The protective role of a moderate protein diet in ETEC-infected piglets: Optimization of growth, immunity, and microbial balance","volume":"25","author":"Li","year":"2026","journal-title":"Anim. Nutr."},{"key":"ref_2","first-page":"71","article-title":"Light and scanning electron microscopy evaluation of the postnatal small intestinal mucosa development in pigs","volume":"56","author":"Skrzypek","year":"2005","journal-title":"J. Physiol. Pharmacol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"6012","DOI":"10.1038\/s41598-022-10059-5","article-title":"Influence of Chlorella vulgaris on growth, digestibility and gut morphology and microbiota of weaned piglet","volume":"12","author":"Martins","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"skad401","DOI":"10.1093\/jas\/skad401","article-title":"Dietary butyrate and valerate glycerides impact diarrhea severity and immune response of weaned piglets under ETEC F4-ETEC F18 coinfection conditions","volume":"101","author":"Kovanda","year":"2023","journal-title":"J. Anim. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Kim, K., Song, M., Liu, Y., and Ji, P. (2022). Enterotoxigenic Escherichia coli infection of weaned pigs: Intestinal challenges and nutritional intervention to enhance disease resistance. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.885253"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1038\/s41522-024-00514-8","article-title":"Protecting the piglet gut microbiota against ETEC-mediated post-weaning diarrhoea using specific binding proteins","volume":"10","author":"Jenkins","year":"2024","journal-title":"npj Biofilms Microbiomes"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Xu, J., Jia, Z., Xiao, S., Long, C., and Wang, L. (2023). Effects of Enterotoxigenic Escherichia coli Challenge on Jejunal Morphology and Microbial Community Profiles in Weaned Crossbred Piglets. Microorganisms, 11.","DOI":"10.3390\/microorganisms11112646"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1186\/s40104-025-01172-2","article-title":"Impacts of zinc caproate supplementation on growth performance, intestinal health, anti-inflammatory activity, and Zn homeostasis in weaned piglets challenged with Escherichia coli K88","volume":"16","author":"Xu","year":"2025","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"5024","DOI":"10.1038\/s41598-022-08446-z","article-title":"Effect of chronic and acute enterotoxigenic E. coli challenge on growth performance, intestinal inflammation, microbiome, and metabolome of weaned piglets","volume":"12","author":"Boeckman","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Wang, Y., Zhang, Z., Du, M., Ji, X., Liu, X., Zhao, C., Pang, X., Jin, E., Wen, A., and Li, S. (2024). Berberine alleviates ETEC-induced intestinal inflammation and oxidative stress damage by optimizing intestinal microbial composition in a weaned piglet model. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1460127"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e0374422","DOI":"10.1128\/spectrum.03744-22","article-title":"Impact of Intestinal Microbiota on Growth Performance of Suckling and Weaned Piglets","volume":"11","author":"Mahmud","year":"2023","journal-title":"Microbiol. Spectr."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Bin, P., Tang, Z., Liu, S., Chen, S., Xia, Y., Liu, J., Wu, H., and Zhu, G. (2018). Intestinal microbiota mediates Enterotoxigenic Escherichia coli-induced diarrhea in piglets. BMC Vet. Res., 14.","DOI":"10.1186\/s12917-018-1704-9"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1016\/j.aninu.2017.10.001","article-title":"Intestinal challenge with enterotoxigenic Escherichia coli in pigs, and nutritional intervention to prevent postweaning diarrhea","volume":"3","author":"Sun","year":"2017","journal-title":"Anim. Nutr."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e11","DOI":"10.1017\/anr.2026.10029","article-title":"Antimicrobial peptides produced by Clostridium butyricum alleviate LPS-induced intestinal injury in piglet by modulating gut microbiota, bile acid, and GPR43-NLRP3 pathway","volume":"3","author":"Chen","year":"2026","journal-title":"Anim. Nutr."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"367","DOI":"10.24425\/pjvs.2023.145042","article-title":"Andrographolide loaded montmorillonite attenuated enterotoxigenic Escherichia coli induced intestinal barrier injury and inflammation in a mouse model","volume":"26","author":"Wang","year":"2023","journal-title":"Pol. J. Vet. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.clay.2013.11.009","article-title":"The effect of dietary bentonite on post-weaning diarrhoea, growth performance and blood parameters of weaned piglets","volume":"90","author":"Trckova","year":"2014","journal-title":"Appl. Clay Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"118405","DOI":"10.1016\/j.lfs.2020.118405","article-title":"Palmatine regulates bile acid cycle metabolism and maintains intestinal flora balance to maintain stable intestinal barrier","volume":"262","author":"Ning","year":"2020","journal-title":"Life Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1614","DOI":"10.5713\/ajas.2012.12698","article-title":"Effects of graded levels of montmorillonite on performance, hematological parameters and bone mineralization in weaned pigs","volume":"26","author":"Duan","year":"2013","journal-title":"Asian-Australas. J. Anim. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"105608","DOI":"10.1016\/j.psj.2025.105608","article-title":"The effects of copper-loaded montmorillonite on intestinal morphology, microbiota, barrier function, antioxidant capacity, and gut-related gene expression in broilers","volume":"104","author":"Hu","year":"2025","journal-title":"Poult. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"21369","DOI":"10.1039\/C8RA03651A","article-title":"Study on montmorillonite\u2013chlorhexidine acetate\u2013terbinafine hydrochloride intercalation composites as drug release systems","volume":"8","author":"Sun","year":"2018","journal-title":"RSC Adv."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Zhang, Y., Tong, Y., Li, P., Zhu, L., Zhang, H., Xie, H., Nasir, S., Li, W., Fang, M., and Wang, J. (2025). Palmatine potentiates cefquinome efficacy against multidrug-resistant Escherichia coli via sulfur\/taurine metabolism and oxidative stress modulation. Front. Microbiol., 16.","DOI":"10.3389\/fmicb.2025.1644399"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"115110","DOI":"10.1016\/j.intimp.2025.115110","article-title":"Palmatine ameliorates intestinal epithelial barrier injury in ulcerative colitis via targeting enolase 3","volume":"162","author":"Zhang","year":"2025","journal-title":"Int. Immunopharmacol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"290","DOI":"10.3109\/00498254.2010.529180","article-title":"Intestinal absorption mechanisms of berberine, palmatine, jateorhizine, and coptisine: Involvement of P-glycoprotein","volume":"41","author":"Zhang","year":"2011","journal-title":"Xenobiotica"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"105491","DOI":"10.1016\/j.clay.2020.105491","article-title":"Purification of montmorillonite and the influence of the purification method on textural properties","volume":"187","author":"Qiao","year":"2020","journal-title":"Appl. Clay Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/j.cemconres.2018.11.015","article-title":"Influence of experimental procedure on d-spacing measurement by XRD of montmorillonite clay pastes containing PCE-based superplasticizer","volume":"116","author":"Borralleras","year":"2019","journal-title":"Cem. Concr. Res."},{"key":"ref_26","unstructured":"NRC (2012). Nutrient Requirements of Swine: Eleventh Revised Edition, The National Academies Press."},{"key":"ref_27","unstructured":"AOAC International (2019). Loss on Drying (Moisture) at 95\u2013100 \u00b0C for Feeds: AOAC Official Method 934.01. Official Methods of Analysis of AOAC International (21st ed.), AOAC International."},{"key":"ref_28","unstructured":"AOAC International (2006). Protein (crude) in animal feed, combustion method: AOAC official method 990.03. Official Methods of Analysis of AOAC International (18th ed., Rev. 1), AOAC International."},{"key":"ref_29","unstructured":"AOAC International (2019). Fat (crude) or ether extract in animal feed: AOAC official method 920.39 (A). Official methods of analysis of AOAC International (21st ed.), AOAC International."},{"key":"ref_30","unstructured":"(2022). Animal Feeding Stuffs\u2014Determination of Crude Ash (Standard No. ISO 5984:2022)."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1017\/S1751731114000068","article-title":"A comparison of two methods for determining titanium dioxide marker content in broiler digestibility studies","volume":"8","author":"Morgan","year":"2014","journal-title":"Animal"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3594","DOI":"10.1016\/j.psj.2020.04.007","article-title":"Effects of dietary vitamins supplementation level on the production performance and intestinal microbiota of aged laying hens","volume":"99","author":"Gan","year":"2020","journal-title":"Poult. Sci."},{"key":"ref_34","unstructured":"Coates, J. (2006). Interpretation of Infrared Spectra, A Practical Approach. Encyclopedia of Analytical Chemistry, John Wiley & Sons Ltd."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1186\/2049-1891-4-52","article-title":"Escherichia coli challenge and one type of smectite alter intestinal barrier of pigs","volume":"4","author":"Almeida","year":"2013","journal-title":"J. Anim. Sci. Biotechnol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"678","DOI":"10.1111\/jpn.13522","article-title":"Dietary copper\/zinc-loaded montmorillonite improved growth performance and intestinal barrier and changed gut microbiota in weaned piglets","volume":"105","author":"Li","year":"2021","journal-title":"J. Anim. Physiol. Anim. Nutr."},{"key":"ref_37","first-page":"2943","article-title":"Excess of dietary montmorillonite impairs growth performance, liver function, and antioxidant capacity in starter pigs","volume":"95","author":"Zhao","year":"2017","journal-title":"J. Anim. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Shi, Z., Han, L., Yang, H., Lu, G., Shi, Z., and Ma, B. (2025). From traditional remedy to modern therapy: A comprehensive review of palmatine\u2019s multi-target mechanisms and ethnopharmacological potential. Front. Pharmacol., 16.","DOI":"10.3389\/fphar.2025.1624353"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"9090","DOI":"10.1038\/s41598-023-36137-w","article-title":"Influence of inorganic and organic salts on the hydration mechanism of montmorillonite based on molecular simulation","volume":"13","author":"Yang","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.clay.2017.07.034","article-title":"Drug release material hosted by natural montmorillonite with proper modification","volume":"148","author":"Wu","year":"2017","journal-title":"Appl. Clay Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.clay.2005.10.010","article-title":"Adsorption and antibacterial effect of copper-exchanged montmorillonite on Escherichia coli K88","volume":"31","author":"Hu","year":"2006","journal-title":"Appl. Clay Sci."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Wang, Q., Zhan, X., Wang, B., Wang, F., Zhou, Y., Xu, S., Li, X., Tang, L., Jin, Q., and Li, W. (2022). Modified Montmorillonite Improved Growth Performance of Broilers by Modulating Intestinal Microbiota and Enhancing Intestinal Barriers, Anti-Inflammatory Response, and Antioxidative Capacity. Antioxidants, 11.","DOI":"10.3390\/antiox11091799"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Liu, H., Wang, C., Gu, X., Zhao, J., Nie, C., Zhang, W., and Ma, X. (2020). Dietary Montmorillonite Improves the Intestinal Mucosal Barrier and Optimizes the Intestinal Microbial Community of Weaned Piglets. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.593056"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Gao, Q., Zhang, Y., Wu, Y., Gu, D., Chen, J., Yin, C., Wu, H., Zhu, D., Chen, D., and Wu, A. (2025). Dietary Fe-Gly supplementation attenuates enterotoxigenic Escherichia coli (ETEC)-induced inflammation response and intestinal barrier dysfunction in piglets. Front. Vet. Sci., 12.","DOI":"10.3389\/fvets.2025.1537604"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"8865503","DOI":"10.1155\/tbed\/8865503","article-title":"Enterotoxigenic Escherichia coli as a Modulator of the Entero-Pulmonary Axis in Piglets: Impacts on the Microbiota and Immune Responses","volume":"2025","author":"Breyer","year":"2025","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1186\/s42523-020-0020-4","article-title":"Late weaning is associated with increased microbial diversity and Faecalibacterium prausnitzii abundance in the fecal microbiota of piglets","volume":"2","author":"Massacci","year":"2020","journal-title":"Anim. Microbiome"},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Jin, W., Zhang, M., Lan, X., Huang, Y., Bai, Y., Li, Y., Shi, C., Song, Y., Wang, L., and Zhang, Y. (2025). Probiotic Weissella cibaria LAB_Weis_Camel_L4 mitigates Escherichia coli-induced enteritis via competitive exclusion and microbiota modulation. Front. Immunol., 16.","DOI":"10.3389\/fimmu.2025.1642209"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/18\/2868\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,13]],"date-time":"2026-09-13T04:10:49Z","timestamp":1789272649000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/18\/2868"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,11]]},"references-count":47,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ani16182868"],"URL":"https:\/\/doi.org\/10.3390\/ani16182868","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,11]]}}}