Bacterial resistance to antibiotics and residues of antibiotics in poultry products have encouraged the use of probiotics, prebiotic substrates, and synbiotic combinations of prebiotics and probiotics as alternative approaches to the use of antibiotics in poultry. The present study was carried out to evaluate the effect of a new probiotic CS61 culture on growth performance, feed conversion efficiency, and safety in broiler chickens, and to evaluate its value as an alternative for antibiotics used as a feed additive. Two dosages of CS61 culture (0.1% and 1%) were fed to chickens for 28 days. The results showed that terminal body weight and daily weight gain in the treatment groups increased in a dose-dependent manner when compared with the control group. Dietary supplementation with CS61 culture also improved feed conversion rate compared to the control group. There were no treatment-related toxic effects in terms of clinical findings, mortality, necropsy findings, hematology, or serum biochemistry parameters in any group tested. The nitric oxide assay showed that CS61 peptide has a dose-dependent inhibitory effect on lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells. The results of this experiment indicated that dietary supplementation of CS61 culture may improve growth performance and feed conversion efficiency in chickens through its anti-inflammatory effect.