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In laying hens, feather-pecking behaviour is proposed to be a consequence of gut-brain axis dysregulation. <jats:italic>Lactobacillus rhamnosus<\/jats:italic> decreases stress-induced severe feather pecking in adult hens, but whether its effect in pullets is more robust is unknown. Consequently, we investigated whether early-life, oral supplementation with a single <jats:italic>Lactobacillus rhamnosus<\/jats:italic> strain can prevent stress-induced feather-pecking behaviour in chickens. To this end, we monitored both the short- and long-term effects of the probiotic supplement on behaviour and related physiological parameters. We hypothesized that <jats:italic>L. rhamnosus<\/jats:italic> would reduce pecking behaviour by modulating the biological pathways associated with this detrimental behaviour, namely aromatic amino acid turnover linked to neurotransmitter production and stress-related immune responses. We report that stress decreased the proportion of cytotoxic T cells in the tonsils (P\u2009=\u20090.047). Counteracting this T cell depression, birds receiving the <jats:italic>L. rhamnosus<\/jats:italic> supplementation significantly increased all T lymphocyte subset proportions (P\u2009&lt;\u20090.05). Both phenotypic and genotypic feather peckers had lower plasma tryptophan concentrations compared to their non-pecking counterparts. The probiotic supplement caused a short-term increase in plasma tryptophan (P\u2009&lt;\u20090.001) and the TRP:(PHE\u2009+\u2009TYR) ratio (P\u2009&lt;\u20090.001). The administration of stressors did not significantly increase feather pecking in pullets, an observation consistent with the age-dependent onset of pecking behaviour. Despite minimal changes to behaviour, our data demonstrate the impact of <jats:italic>L. rhamnosus<\/jats:italic> supplementation on the immune system and the turnover of the serotonin precursor tryptophan. Our findings indicate that <jats:italic>L. rhamnosus<\/jats:italic> exerts a transient, beneficial effect on the immune response and tryptophan catabolism in pullets.<\/jats:p>","DOI":"10.1038\/s41598-021-98459-x","type":"journal-article","created":{"date-parts":[[2021,10,1]],"date-time":"2021-10-01T11:02:54Z","timestamp":1633086174000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["L. rhamnosus improves the immune response and tryptophan catabolism in laying hen pullets"],"prefix":"10.1038","volume":"11","author":[{"given":"Claire","family":"Mindus","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nienke","family":"van Staaveren","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dietmar","family":"Fuchs","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna M.","family":"Gostner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joergen B.","family":"Kjaer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wolfgang","family":"Kunze","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. 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