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This study examined the effects of adding lysozyme and\n                    <jats:italic>Bacillus subtilis<\/jats:italic>\n                    on growth indices, lipid profiles, immune responses, gut integrity, and gene expression in heat-stressed broilers. A total of 600 one-day-old (Ross 308) male chicks were randomly allocated into four treatment groups (six replicates) as follows: CON, a basal diet without feed additives; B.S, a basal diet supplemented with\n                    <jats:italic>B. subtilis<\/jats:italic>\n                    (500\u00a0mg\/kg diet); LYS, a basal diet supplemented with lysozyme (150\u00a0mg\/kg diet); and BSLY, a basal diet supplemented with both\n                    <jats:italic>B. subtilis<\/jats:italic>\n                    and lysozyme (500\u00a0mg\/kg and 150\u00a0mg\/kg diet, respectively). Chickens were exposed to a temperature of 33\u00a0\u00b0C for 4\u00a0h daily during the experimental period. Results indicated that broilers receiving a diet supplemented with BSLY showed higher body weight gain and lower feed conversion ratio than the control (\n                    <jats:italic>p<\/jats:italic>\n                    \u2009&lt;\u20090.05). The carcass characteristics and nutrient digestibility of the BSLY group were also improved. Total cholesterol decreased in chickens fed BSLY and B.S, while low-density lipoprotein (LDL) levels decreased in the LYS group. compared to the control group; however, high-density lipoprotein (HDL) increased in the BSLY, B.S, and LYS groups. Additionally, the addition of BSLY and B.S enhanced the immune response by increasing the levels of immunoglobulin G (IgG), immunoglobulin M (IgM), and the relative weight of the Bursa of Fabricius. Furthermore, gut integrity improved via expanding the ileum villus height in chickens fed BSLY, B.S, and LYS , as well as the counts of\n                    <jats:italic>Bacillus<\/jats:italic>\n                    spp. and\n                    <jats:italic>Lactobacillus<\/jats:italic>\n                    in the broilers fed BSLY and B.S. Dietary BSLY, B.S, and LYS modulated gene expression, via up-regulation of insulin-like growth factor 1 (IGF1), superoxide dismutase (SOD), glutathione peroxidase (GPx), and interferon\u2010gamma (IFN-\u03b3) gene expression compared to the control group, while the highest SOD and IGF1 genetic modification was in the group that received BSLY. In conclusion, adding\n                    <jats:italic>B. subtilis<\/jats:italic>\n                    -lysozyme mixture can have a positive impact on growth performance indices, immune response, gut health, and gene expression, thereby increasing heat stress resistance of broiler chickens.\n                  <\/jats:p>","DOI":"10.1038\/s41598-025-23910-2","type":"journal-article","created":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T15:30:26Z","timestamp":1762961426000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Lysozyme and Bacillus subtilis improve growth performance, nutrient utilization, physiological stress indices, intestinal integrity, and gene expression of heat-stressed broilers"],"prefix":"10.1038","volume":"15","author":[{"given":"Ahmed M.","family":"Elbaz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"G. 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