This study evaluated the efficacy of Barnouf (Pluchea indica Less) extract (BE) as a natural antioxidant and immune-modulating supplement in broiler chickens. A total of 210 chicks were assigned to three treatments in a completely randomized design, with each treatment consisting of seven replicates of ten unsexed chicks. The control group received water without additives, whereas the other treatments received Barnouf extract (BE) at 3 or 6 mL/L. Supplementation with BE at both inclusion levels improved growth performance (P ≤ 0.05). Birds receiving 6 mL/L exhibited greater body weight gain (BWG) from days 5 to 20 and days 21 to 33 compared with the control. Feed intake (FI) decreased (P ≤ 0.05) in both supplemented groups, accompanied by improved feed conversion ratio (FCR); additionally, the 3 mL/L group demonstrated a higher performance index (PI) (P ≤ 0.05) on day 33. Broilers receiving 3 or 6 mL/L of the extract showed increased water consumption (WC) and higher water-to-feed ratios between days 5 and 20 (P ≤ 0.05), with the 6 mL/L group showing the greatest values, while still exhibiting reduced FI. Supplementation with BE increased abdominal fat percentage, live body weight, and relative weights of the gizzard and liver, whereas carcass percentage varied among treatments. Heart and spleen weights showed numerical reductions (P ≥ 0.05) with increasing extract levels. Health-related biochemical indices were favorably altered by BE supplementation. Levels of aspartate aminotransferase (AST), urea, total lipids, and malondialdehyde (MDA) were reduced (P ≤ 0.05), whereas concentrations of high-density lipoprotein (HDL), antioxidant enzymes—glutathione peroxidase (GPX), glutathione S-transferase (GST), and reduced glutathione (GSH)—and immunoglobulins G (IgG) and M (IgM) increased (P ≤ 0.05). Meat quality also improved, as reflected by more stable pH values, lower microbial loads, and reduced storage-related water loss, with significant effects observed for color parameters (P ≤ 0.05). Additionally, both supplementation levels increased lactobacilli count while reducing total bacterial count (TBC), bacilli, Escherichia coli, coliforms, and Salmonella (P ≤ 0.05). In conclusion, dietary inclusion of BE at 3 or 6 mL/L enhanced growth performance, immune competence, antioxidant status, meat quality, and gut microbial balance in broiler chickens without adverse effects, supporting its potential as a safe natural feed additive.
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