At present, with the development of research on substitute products for feed antibiotics, there is a diversification of varieties and functions of substitute antibiotics in the market, which have certain effects in promoting the growth and health of livestock and poultry, improving the quality of livestock products, etc. The main antibiotic substitutes in livestock and poultry breeding include probiotics, antimicrobial peptides, enzyme preparations, polysaccharides, and plant extracts. The evaluation methods for these antibiotic alternatives include: ① using omics techniques (genomics, proteomics, transcriptomics, etc.) to elucidate the mechanism of action, determine the multi-target effects of antibiotic alternatives on animals, explore the main and auxiliary functions of alternatives, and thus better match different alternatives; ② Optimize conventional animal production evaluation methods, establish multiple in vitro models for antibiotic substitution technology research, develop a safety evaluation method system that combines in vivo animal models with in vitro detection techniques, screen early biomarkers and sensitive markers, establish a systematic evaluation of effectiveness, safety, and healthy growth performance; ③ Evaluate antibiotic alternatives based on diagnosis, prognosis, efficacy, and safety, measure their dynamic changes in vivo content using high-throughput experimental methods, and attempt to simulate the effects of these changes on cells, tissues, organs, and the overall animal body by establishing mathematical and computer models.