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This study aimed to compare novel multi-strain and single-strain <jats:italic>Bacillus<\/jats:italic> probiotics and their respective influences on broiler chickens\u2019 performance, gut health, litter quality, immune response, and <jats:italic>NBN<\/jats:italic> and <jats:italic>TLR<\/jats:italic> gene expression. A total of 1200 Arbor-Acres 1-day-old broiler chicks were randomly allocated into three treatments (T1 was a control, T2 was supplemented with a combined <jats:italic>Bacillus coagulans<\/jats:italic> (2\u2009\u00d7\u200910<jats:sup>9<\/jats:sup>\u00a0cfu\/g) and <jats:italic>Bacillus licheniformis<\/jats:italic> (8\u2009\u00d7\u200910<jats:sup>9<\/jats:sup>\u00a0cfu\/g) probiotic strains (0.2\u00a0kg\/ton of feed), and T3 was supplemented with <jats:italic>Bacillus licheniformis<\/jats:italic> (3.2\u2009\u00d7\u200910<jats:sup>9<\/jats:sup>\u00a0cfu\/g) probiotic (0.5\u00a0kg\/ton of feed) with eight replicas of each. Supplementing the broiler diet with either the single-strain (T3) or the multi-strain (T2) <jats:italic>Bacillus<\/jats:italic>-based probiotic raised the overall birds\u2019 body weight, body weight gain, feed conversion ratio, and European production efficiency factor compared to the control (T1), with a significant enhancement achieved by the multi-strain <jats:italic>Bacillus<\/jats:italic> product (<jats:italic>P<\/jats:italic>\u2009=\u20090.005). T2 and T3 exhibited significantly improved cholesterol, Alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and alkaline phosphatase levels than the control (<jats:italic>P<\/jats:italic>\u2009\u2264\u20090.05). The transcript levels of both <jats:italic>NBN<\/jats:italic> and <jats:italic>TLR<\/jats:italic> genes were upregulated in the liver in the T2 and T3 groups. The T2 group experienced significant reductions in gut bacterial counts, especially for <jats:italic>Clostridia<\/jats:italic>, and recorded the lowest litter moisture and nitrogen. In conclusion, supplementing broiler diets with probiotics of multiple <jats:italic>Bacillus<\/jats:italic> strains increased production profitability by promoting bird growth, improving feed intake, enhancing gut mucosa and immune organs, and upregulating genes responsible for immunity. All these inhibit the overgrowth of enteric pathogens and sustain litter quality.<\/jats:p>","DOI":"10.1007\/s11250-023-03453-2","type":"journal-article","created":{"date-parts":[[2023,1,14]],"date-time":"2023-01-14T05:02:40Z","timestamp":1673672560000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["Influence of dietary Bacillus coagulans and\/or Bacillus licheniformis-based probiotics on performance, gut health, gene expression, and litter quality of broiler chickens"],"prefix":"10.1007","volume":"55","author":[{"given":"Ebtihal M. 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