Microbial adhesion and biofilm formation on surfaces pose major problems and risks to human health. One way to circumvent this problem is to coat surfaces (in this report stainless steel) with a non-toxic fish extract that generates an abiotic surface with less bacterial attachment than uncoated surfaces or surfaces coated with, for example, tryptone soy broth. The bacteria grow well in the fish extract; hence a general bacteriocidal effect is not the reason for the antifouling effect. Bacterial attachment was quantified by different methods including (a) direct fluorescence microscopy, (b) removal by ultrasound and subsequent quantification of the adhered bacteria, and (c) regrowth of the adhered bacteria measured by indirect conductometry. Surprisingly, the bacterial counts on surfaces coated with aqueous fish extract were 10–100 times lower than on surfaces coated with laboratory broths when surfaces were submerged in bacterial suspensions. The effect was seen for Pseudomonas fluorescens AH2, Pseudomonas aeruginosa PAO1, Escherichia coli MG1655, Vibrio anguillarum 90-11-287 and Aeromonas salmonicida Jno 3175/88. It lasted for at least 7 days. Atomic force microscopy showed that steel surfaces conditioned with fish extract were covered by a thin layer of spherical, nanosized particles. Chemical analysis of the surfaces coated with adsorbed fish extract using X-ray photoelectron spectroscopy revealed that the layer was proteinaceous and had a thickness less than 2 nm. Numerous protein bands/peaks were also detected by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry techniques. We conclude that coating the stainless steel surface with fish extract results in a thin protein layer that reduces bacterial adhesion significantly.