This study evaluated the effects of chemical biocides used to control dental unit waterline biofilm on the bond strength of resin to enamel. Sixty bovine teeth were randomly assigned to six treatment groups. One-way ANOVA revealed a significant difference in means (p < 0.001) and Tukey's multiple range test indicated that three of the experimental groups had significantly lower mean shear bond strengths than the control (p < 0.05). This finding suggests that dental unit waterline biocides may adversely affect adhesion of resin to enamel.