Background: Proliferation of bacteria, such as Staphylococcus aureus and Staphylococcus epidermidis, on orthopedic implants has been a challenge in orthopedic surgery, highlighting the problem of the increas-ing antibiotic resistance and the necessity to develop new antimicrobial agents. In this sense, antimicro-bial peptides are promising candidates, which can be attached to titanium surfaces in order to make them safer. Mytilus mussels are characterized by adhering efficiently to a wide variety of surfaces, especially metallic ones, through adhesive proteins with a high content of dihydroxyphenylalanine, which is a post-translational modification of tyrosine. Results: This work refers to the synthesis of a coating based on a bifunctional peptide that combines a sequence derived from the mussel foot protein-5 and the antimicrobial peptide pEM-2, derived from Bothrops asper snake venom. The adhesive properties of this bifunctional peptide were evaluated, as well as the adhesive sequence without the pEM-2, using a Quartz Crystal Microbalance. The results showed that the presence of the antimicrobial peptide improved the adhesion; however, a loss of the bactericidal activity was observed. Even so, the adhesive sequence by itself exhibited an important antifouling activ-ity, preventing S. aureus and S. epidermidis adhesion to titanium by 75% and 45%, respectively, although the result against S. epidermidis was not significant. Conclusions: A 13-residue peptide derived from a natural and biocompatible source, like a Mytilus mussel adhesive protein, could be projected as a protective agent on titanium surfaces against S. aureus and S. epidermidis, being responsible for two thirds of the cases of orthopedic implant infection. How to cite: Gauna A, Mercado L, Guzman F. Antimicrobial characterization of a titanium coating derived from mussel-glue and Bothrops asper snake venom for the prevention of implant-associated infections caused by Staphylococcus. Electron J Biotechnol 2022;56. https://doi.org/10.1016/j.ejbt.2022.02.001. (c) 2022 Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).