The skins of frogs of the family Ranidae are particularly rich sources of biologically active pep-tides,among which antimicrobial peptides (AMPs) constitute the major portion.Some of these have attracted the interest of researchers because they possess both antimicrobial and anticancer activities.In this study,with\'shotgun\'cloning and MS/MS fragmentation,three AMPs,homo-Iogues of family brevinin-1 (brevinin-1HL),and temporin (temporin-HLa and temporin-HLb),were discovered from the skin secretion of the broad-folded frog,Hylarana/atouchii.They exhibited various degrees of antimicrobial and antibiofilm activities against test microorganisms and hemol-ysis on horse erythrocytes.It was found that they could induce bacteria death through disrupting cell membranes and binding to bacterial DNA.In addition,they also showed different poten-cies towards human cancer cell lines.The secondary structure and physicochemical properties of each peptide were investigated to preliminarily reveal their structure-activity relationships.Cir-cular dichroism spectrometry showed that they all adopted a canonical α-helical conformation in membrane-mimetic solvents.Notably,the prepropeptide of brevinin-1HL from H.latouchii was highly identical to that of brevinin-1GHd from Hylarana guentheri,indicating a close relationship between these two species.Accordingly,this study provides candidates for the design of novel anti-infective and antineoplastic agents to fight multidrug-resistant bacteria and malignant tumors and also offers additional clues for the taxonomy of ranid frogs.