The microbiological load and microbiome of the Dutch dental unit; 'please, hold your breath'.

Dental unit water systems are prone to biofilm formation. During use of the dental unit, clumps of biofilm slough off and can subsequently be aerosolized and inhaled by both patient and staff, potentially causing infections. The aim of this study was to determine the microbial load and microbiome of dental unit water, in the Netherlands, and the factors influencing these parameters. In total, 226 dental units were sampled and heterotrophic plate counts (HPC) were determined on the traditional effluent sample. Of all dental units, 61% exceeded the recommended microbiological guidelines of 100 colony forming units per milliliter. In addition, the microbiome, with additional q-PCR analysis for specific species, was determined on an effluent sample taken immediately after an overnight stagnancy period, in which the biofilm is in its relaxed state. These relaxed biofilm samples showed that each dental unit had a unique microbiome. Legionella spp., amoeba and fungi were found in 71%, 43% and 98% of all units, respectively. The presence of amoeba was positively associated with nine bacterial biomarkers and correlated positively with bacterial and fungal DNA and Legionella spp. concentrations, but not with HPC. Only when adhering to disinfection protocols, statistically significant effects on the microbial load and microbiome were seen. The relaxed biofilm sample, in combination with molecular techniques gives better insight in the presence of opportunistic pathogens when compared to the heterotrophic plate counts. Infection control measures should focus on biofilm analysis and control in order to guarantee patient safety.