The invention relates to the technical field of protein engineering, in particular to a multifunctional hybrid peptide with antibacterial, anti-inflammatory, endotoxin neutralizing and immunomodulatory activities as well as a preparation method and application thereof. The hybrid peptide provided by the invention comprises a peptide I and a peptide II fused with the peptide I, wherein an amino acid sequence of the peptide I is as shown in SEQ ID NO.1, and an amino acid sequence of the peptide II is as shown in 19th-29th sites of SEQ ID NO.2. The hybrid peptide has antibacterial, endotoxin neutralizing, immunoregulation and anti-inflammatory functions, has the advantages of low cytotoxicity, high safety, convenience in preparation and low cost, can be used as an ideal antibacterial agent, endotoxin neutralizing agent, immunoregulator and anti-inflammatory agent, is widely applied to the fields of medicines, foods, health care, feeds, nutrition and the like of human beings and animals, and has good application potential and value.