Workers examining the transmission route(s) and reservoir(s) of infection for Helicobacter pylori have postulated several environmental reservoirs for the organism, including water. Such work has, to date, concentrated on the bulk liquid in drinking water systems rather than on biofilms. Previous investigations by the authors have suggested biofilms in water distribution systems are a possible reservoir of infection. This current study comprised of an analysis of a section of cast iron mains distribution pipe removed from an urban environment in the north-east of Scotland during routine maintenance work. Immediately upon removal of the pipe section, the interior lumen was swabbed to remove the biofilm layer. Subsequent analysis for the presence of Helicobacter DNA using a nested PCR approach produced a positive result. This data provides the first evidence for the existence of Helicobacter in biofilms found in water distribution systems anywhere in the world.