Evaluation of power ultrasound for disinfection of both Legionella pneumophila and its environmental host Acanthamoeba castellanii

The objectives of this study were to (1) examine the effect of power ultrasound on the viability of both Legionella pneumophila and Acanthamoeba castellanii trophozoites and cysts, (2) investigate if intracellular Legionella replication in trophozoites positively affects bacterial resistance to ultrasound and (3) study if Legionella renders viable but non-culturable (VBNC) due to ultrasound treatments. Using laboratory scale experiments, microorganisms were exposed for various time periods to power ultrasound at a frequency of 36 kHz and an ultrasound power setting of 50 and 100%. Due to a fast destruction, trophozoite hosts were not able to protect intracellular Legionella from eradication by ultrasound, in contrast to cysts. No significant effects of ultrasound on cyst viability could be detected and power settings of 100% for 30 min only made intracellular Legionella concentrations decrease with 1.3 log units. Due to intracellular replication of Legionella in trophozoites, ultrasound no longer affected bacterial viability. Concerning the VBNC state, ultrasound treatments using a power setting of 50% partly induced Legionella (±7%) to transform into VBNC bacteria, in contrast to power settings of 100%. Promising results obtained in this study indicate the possible application of power ultrasound in the control of both Legionella and Acanthamoeba concentrations in anthropogenic water systems.