Legionella Biofilm Formation Potential on Different Pipe Materials Used for Low Nutrient Municipality Tap Water

Factors affecting biofilm development, specifically the materials of the pipe, were investigated. Two laboratory scale bioreactor systems were constructed to study biofilm formations: a pipe loop bioreactor with continuous flow at 10.1 liters per minute (LPM), and a tank bioreactor under stagnant conditions with a minimal flow of 0.0095 LPM. The continuous flow bioreactors were constructed using cross-linked polyethylene (PEX), copper, and galvanized steel pipes. The tank bioreactors consisted of glass chambers containing coupons made from the pipe materials, as well as glass microscope slides. Municipality tap water was used in the experimentation, with no nutrients added. Legionella pneumophila was spiked into all the pipe loop bioreactors, and only in one tank bioreactor. Detection of heterotrophic bacteria, coliforms and Legionella using tryptic soy agar (TSA), Brilliance, and buffered yeast charcoal extract (BYCE), respectively. Over ten weeks, biofilms were developed on PEX, copper, and steel, in the pipe loop bioreactors and the tank bioreactors. Heterotrophic bacteria were detected in all systems; however, no coliforms were detected, and Legionella pneumophila was only detected on a coupon in the copper pipe loop bioreactor, as measured by bacterial concentration on test materials. In the tank bioreactors, biofilms developed the most rapidly on PEX, followed by galvanized steel, and finally copper. Out of the four materials, copper had the lowest bacterial growth, which can be ascribed to the bactericidal impact of copper ions on the bacterial cells attaching to the copper surface. After biofilm aging, higher bacterial colonization on copper and accumulation of dead bacterial layer on the surface may act as a protective barrier against copper ions. Bacterial densities in the biofilm reached a high concentration of 1.40 x 105 CFU/cm2 on the PEX pipe loop bioreactor, and 1.05 x 104 CFU/cm2in the PEX coupon in the tank bioreactors. Comparing the turbulent conditions in the pipe loop bioreactors to the stagnant conditions in the tank bioreactor, showed that biofilms formed more rapidly under stagnant conditions, but in larger quantities under turbulent conditions.