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Columnaris disease in channel catfish, <jats:italic>Ictalurus punctatus<\/jats:italic>, is primarily caused by <jats:italic>Flavobacterium covae<\/jats:italic> (formerly <jats:italic>F.\u2009columnare<\/jats:italic>). Immunostimulants may enhance nonspecific immune responses and offer an alternative to antibiotic treatments in catfish. Furthermore, dietary protein sources and inclusions are also essential to fish health and nutrition and may enhance overall fish performance in pond culture. The current project evaluated two immunostimulants: a protease complex (PC) and a humic substance (HS) derived from a reed\u2010sedge peat product. A 60\u2010day trial examined the effects of five diets on growth performance, immune response and resistance to experimental <jats:italic>F.\u2009covae<\/jats:italic> infection in channel catfish. Diets included a high\u2010quality fishmeal diet (32%; CF32), a high\u2010protein soy\u2010based diet (32%; C32), a low\u2010protein soy\u2010based diet (28%; C28; predominately used in industry), a low\u2010protein soy diet supplemented with C28\u2009+\u2009PC at 175\u2009g metric ton<jats:sup>\u22121<\/jats:sup> and C28\u2009+\u2009HS in a low\u2010protein diet at 23\u2009g metric ton<jats:sup>\u22121<\/jats:sup>. Following feeding for 60 d, juvenile channel catfish were sampled for growth performance and baseline health indicators (<jats:italic>n<\/jats:italic>\u2009=\u20093; body mucus, blood for sera, kidney and spleen). A subset of fish was then subjected to an immersion\u2010based in vivo challenge trial with <jats:italic>F.\u2009covae<\/jats:italic> (ALG\u201000\u2010530; 10<jats:sup>6<\/jats:sup>\u2009CFU\u2009mL<jats:sup>\u22121<\/jats:sup> exposure). At 60d post\u2010initiation, there were no dietary differences in the relative growth rate (<jats:italic>p<\/jats:italic>\u2009=\u2009.063) or thermal growth coefficient (<jats:italic>p<\/jats:italic>\u2009=\u2009.055), but the 32% diets generally appeared to perform best. Post\u2010challenge, the C32 group's mortality was higher than the C28\u2009+\u2009PC (<jats:italic>p\u2009=<\/jats:italic>\u2009.006) and C28\u2009+\u2009HS diets (<jats:italic>p\u2009=<\/jats:italic>\u2009.005). Although not significant, the C28 and CF32 groups also demonstrated higher mortality compared to both PC and HS diets. Sera lysozyme concentration was found to increase following pathogen challenge (<jats:italic>p<\/jats:italic>\u2009&lt;\u2009.001) and in comparison with mock\u2010challenged catfish (<jats:italic>p<\/jats:italic>\u2009&lt;\u2009.001). Elevated expression levels of proinflammatory cytokines (<jats:italic>il\u20101\u03b2<\/jats:italic>, <jats:italic>il\u20108<\/jats:italic>, <jats:italic>tnf\u2010\u03b1<\/jats:italic> and <jats:italic>tgf\u2010\u03b2<\/jats:italic>) were observed at trial midpoint and post\u2010infection when compared to 60d. The C28 treatment was found to have lower <jats:italic>tnf\u2010\u03b1<\/jats:italic> expression than the C28\u2009+\u2009PC (<jats:italic>p<\/jats:italic>\u2009=\u2009.042) and C28\u2009+\u2009HS (<jats:italic>p<\/jats:italic>\u2009=\u2009.042) groups following exposure to <jats:italic>F.\u2009covae<\/jats:italic>. These challenge data suggest that the immunostimulants (PC and HS) in plant\u2010based protein may be beneficial in protecting against <jats:italic>F.\u2009covae<\/jats:italic> when offered in low\u2010protein channel catfish diets.<\/jats:p>","DOI":"10.1111\/jfd.13835","type":"journal-article","created":{"date-parts":[[2023,7,18]],"date-time":"2023-07-18T01:44:03Z","timestamp":1689644643000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Investigation of dietary exogenous protease and humic substance on growth, disease resistance to <i>Flavobacterium covae<\/i> and immune responses in juvenile channel catfish (<i>Ictalurus punctatus<\/i>)"],"prefix":"10.1111","volume":"48","author":[{"given":"Abdulmalik A.","family":"Oladipupo","sequence":"first","affiliation":[{"name":"School of Fisheries, Aquaculture, and Aquatic Sciences Auburn University  Auburn Alabama USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9657-6594","authenticated-orcid":false,"given":"Anita M.","family":"Kelly","sequence":"additional","affiliation":[{"name":"School of Fisheries, Aquaculture, and Aquatic Sciences Auburn University  Auburn Alabama USA"},{"name":"Alabama Fish Farming Center Auburn University  Greensboro Alabama USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3822-3624","authenticated-orcid":false,"given":"D. 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