This study aimed to demonstrate the bioadhesive characteristics of gelatin biofilm to rat skin. The biofilm was manufactured from gelatin extracted from the acetic acid treated-skin of the yellowfin tuna Thunnus albacares. The bioadhesive strength of tuna gelatin biofilm was compared to that of porcine gelatin biofilm. The tuna gelatin biofilm exhibited a higher bioadhesive strength than the porcine gelatin biofilm. Gelatin biofilm was subjected to glutaraldehyde treatment at different concentrations, temperatures and pH in order to improve its bioadhesive strength. Glutaraldehyde treatment improved the bioadhesive strength of gelatin biofilm up to three-fold. The bioadhesive strength of glutaraldehyde treated-biofilm was significantly decreased by application of sodium borohydride.