This study investigated the effects of puncture vine (Tribulus Terrestris) addition on the performance, carcass and meat characteristics, selected metabolic and immunological blood indicators, some microbiota, and short-chain fatty acids. A total of 252 1-day-old broilers were distributed to three treatments with 12 cages as replicates per treatment (T1 = 0.0%, T2 = 0.08%, and T3 = 0.16% puncture vine). Performance parameters and metabolic and immunological serum indicators were measured in each feeding phase, while carcass characteristics, meat quality, cecal microflora, and short-chain fatty acids were measured at 35 days. Results showed that live weight, weight gain, production efficiency, and meat component color were lower in initial and ultimate at T3, while the percentages of the legs and gizzard were higher at T2 than T1. The relative weight of cooking loss was higher in T2 and T3, but the myofibril fragmentation index was lower than T1. Total protein and globulin were higher in T2 and T3 (14 days old), and the glucose level was lower in T2 (35 days old) than at T1. Interleukins (IL-4, IL-6, and IL-10) and tumor necrosis factor-α (TNF-α) were higher in T2 than T1 and T3 (35 days old). Puncture vine has antimicrobial activity against Escherichia coli and Salmonella spp., while Lactobacillus spp. was higher in T2. The total short-chain fatty acid content was higher in chickens fed puncture vine. These results indicate that the use of puncture vine powder as a natural alternative at a dosage of 0.08% has a safe effect on the performance, carcass and meat characteristics, and health of broilers.