Survival of Mycobacterium avium in drinking water biofilms as affected by water flow velocity, availability of phosphorus, and temperature

Abstract

Mycobacterium avium is a potential pathogen occurring in drinking water systems. It is a slowly growing bacterium producing a thick cell wall containing mycolic acids, and it is known to resist chlorine better than many other microbes. Several studies have shown that pathogenic bacteria survive better in biofilms than in water. By using Propella biofilm reactors, we studied how factors generally influencing the growth of biofilms (flow rate, phosphorus concentration, and temperature) influence the survival of M. avium in drinking water biofilms. The growth of biofilms was followed by culture and DAPI (4',6'-diamidino-2-phenylindole) staining, and concentrations of M. avium were determined by culture and fluorescence in situ hybridization methods. The spiked M. avium survived in biofilms for the 4-week study period without a dramatic decline in concentration. The addition of phosphorus (10 microg/liter) increased the number of heterotrophic bacteria in biofilms but decreased the culturability of M. avium. The reason for this result is probably that phosphorus increased competition with other microbes. An increase in flow velocity had no effect on the survival of M. avium, although it increased the growth of biofilms. A higher temperature (20 degrees C versus 7 degrees C) increased both the number of heterotrophic bacteria and the survival of M. avium in biofilms. In conclusion, the results show that in terms of affecting the survival of slowly growing M. avium in biofilms, temperature is a more important factor than the availability of nutrients like phosphorus.

FIG. 1.

HPC (A), number of total bacteria (B), M. avium plate count (C), and M. avium analyzed by FISH (D) in the effluent waters and biofilms with different flow velocities. Symbols for biofilms: ▪, 0.1 m/s, CFU, or cells/cm²; □, 0.24 m/s, CFU, or cells/cm². Symbols for water: ▴, 0.1 m/s, CFU, or cells/ml; ▵, 0.24 m/s, CFU, or cells/ml; ×, inlet water, CFU, or cells/ml. Error bars indicate standard deviations.

FIG. 2.

HPC (A), number of total bacteria (B), M. avium plate count (C), and M. avium analyzed by FISH (D) in the effluent waters and biofilms with different phosphorus concentrations. Symbols for biofilms: ▪, 4.2 μg/liter, CFU, or cells/cm²; □, 13.8 μg/liter, CFU, or cells/cm². Symbols for water: ▴, 4.2 μg/liter, CFU, or cells/ml; ▵, 13.8 μg/liter, CFU, or cells/ml; ×, inlet water, CFU, or cells/ml. Error bars indicate standard deviations.

FIG. 3.

HPC (A), number of total bacteria (B), M. avium plate count (C), and M. avium analyzed by FISH (D) in the effluent waters and biofilms with different temperatures. Symbols for biofilms: ▪, 7°C, CFU, or cells/cm²; □, 20°C, CFU, or cells/cm². Symbols for water: ▴, 7°C, CFU, or cells/ml; ▵, 20°C, CFU, or cells/ml; ×, inlet water, CFU, or cells/ml. Error bars indicate standard deviations.