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Therefore, the aim of this study is to investigate the effects of dietary supplementation with multispecies probiotic (MSP) on diarrhea, average daily gain (ADG) and intestinal development of neonatal calves challenged with <jats:italic>Escherichia coli<\/jats:italic> K99.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>Thirty\u2010six neonatal Holstein calves were randomly assigned to three treatment groups. After <jats:italic>E. coli<\/jats:italic> K99 challenge, calves in the control (C) and MSP treatment groups had significantly higher ADG and feed efficiency, and significantly lower fecal scores than those of calves in the diarrhea (D) group. The mean time of diarrhea resolution was 4.5 and 3.1\u2009days for calves in the D and MSP treatment groups, respectively. Furthermore, the structures of the various segments (duodenum, jejunum and ileum) of the small intestine of the calves, activities of several small intestinal enzymes, and expression of several energy metabolism\u2010related genes in the small intestine segments were significantly affected by MSP treatments.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSION<\/jats:title><jats:p>Dietary supplementation of MSP had a positive effect in treating calf diarrhea; it improved ADG and feed efficiency and promoted development of the small intestine. \u00a9 2022 The Authors. <jats:italic>Journal of The Science of Food and Agriculture<\/jats:italic> published by John Wiley &amp; Sons Ltd on behalf of Society of Chemical Industry.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jsfa.11791","type":"journal-article","created":{"date-parts":[[2022,1,23]],"date-time":"2022-01-23T06:47:55Z","timestamp":1642920475000},"page":"4373-4383","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Effects of dietary supplementation with multispecies probiotics on intestinal epithelial development and growth performance of neonatal calves challenged with <i>Escherichia coli<\/i> K99"],"prefix":"10.1002","volume":"102","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1451-4603","authenticated-orcid":false,"given":"Yan\u2010yan","family":"Wu","sequence":"first","affiliation":[{"name":"College of Animal Science and Technology Shihezi University  Shihezi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cun\u2010xi","family":"Nie","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Shihezi University  Shihezi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunsheng","family":"Xu","sequence":"additional","affiliation":[{"name":"College of Animal Science and Technology Shihezi University  Shihezi China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rui\u2010qing","family":"Luo","sequence":"additional","affiliation":[{"name":"Xinjiang Tianshan Junken Animal Husbandry Co. 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