Planktonic free-living amoebae susceptibility to dental unit waterlines disinfectants.

A high diversity of microorganisms is encountered inside dental unit waterlines (DUWL). Among those the presence of free-living amoebae (FLA) appears currently underestimated, although human infections may occur due to contact with FLA-contaminated water during dental cares. In order to limit microbial DUWL contamination, disinfectants are provided by dental unit manufacturer, however, with limited documentation on their activities against FLA. The aim of this study was to evaluate the efficiency of three commercial DUWL disinfectants: the Calbenium© (Airel, Champigny-sur-Marne, France), the Oxygenal 6© (Kavo, Biberach, Germany) and the Sterispray© (Gammasonic, Billom, France), against two FLA species, i.e. Acanthamoeba castellanii and Vermamoeba vermiformis alone or co-cultured with Pseudomonas aeruginosa and Candida albicans at concentrations ranging from 0% to 5% (v/v). Results showed varied efficacies of disinfectants: the Oxygenal 6© did not exhibit FLA killing activity, while the Sterispray© and the Calbenium© displayed concentration- and species-dependent activities with a maximum eradication rates of 100% and 86%, and 79% and 97% for A. castellani and V. vermiformis, respectively. None of the disinfectants were able to totally eradicate FLA at concentrations recommended by manufacturers. Present results highlight unsatisfactory anti-FLA activities of 3 DUWL disinfectant preparations advocating deeper investigation of antimicrobial spectra of commercial disinfectants in use for DUWL maintenance.

Efficacy of dental unit disinfectants against Candida spp. and Hartmannella vermiformis.

Human oral commensal Candida yeasts, as well as environmental free-living amoebae (FLA) such as Hartmannella, are known to be direct or indirect human pathogens. These microorganisms may be isolated from dental unit waterlines (DUWL), because of contamination coming from the tap water and/or a patient's mouth. This study compared the efficacy of commonly used DUWL disinfectants (chlorine, H2 O2 , and Oxygenal 6©) against three species of Candida (C. albicans, C. glabrata, and C. parapsilosis) and one FLA species (H. vermiformis), growing either as single or as mixed biofilms in tap water. Results showed variable efficacies: H2 O2 had no significant activity, while chlorine was effective but only at the highest doses tested, probably not compatible with DUWL uses. Oxygenal 6© was the most efficacious in preventing the growth of yeasts in tap water. However, in the presence of FLA, Oxygenal 6© displayed a reduced antimicrobial activity against sessile C. albicans. In conclusion, none of the tested disinfectants could eradicate yeasts or FLA. Moreover, the antiyeast activity of Oxygenal 6© was reduced in the presence of FLA. Both sessile or planktonic and mixed or single-species conditions should be considered when evaluating the activity of disinfectants for DUWL maintenance. This study also highlighted that FLA should be included in the testing protocols.

Free-living Amoebae (FLA): morphological and molecular identification of Acanthamoeba in dental unit water.

UNLABELLED: The aim of our study was to detect free-living Amoebae (FLA) by morphological methods and to identify Acanthamoeba spp. by PCR in the dental unit water lines (DUWL). MATERIALS AND METHODS: it was a prospective study dealing with 196 water samples collected from DUWL (94 samples taken in the early morning before materials flush and patient consultations and 102 samples taken after consultations). At the same time, 39 samples from tap water were realized. RESULTS: 135 (69%) samples were positives by the morphological study with morpholypical diversity. The predominant morphotype was the monopodial (39.2%). 18 strains of Acanthamoeba spp. were detected in DUW (13.%) and three strains in tap water (10%). The amplification of 18S rDNA gene of these strains of Acanthamoeba spp. was positive for all samples. CONCLUSION: the FLA and Acanthamoeba were isolated both in tap water and in dental unit. The amoeba pathogenicity has not been demonstrated after oral or dental contamination; but the presence of intracellular and pathogenic bacteria in the amoeba could be a source of microbiological risks for patients in case of deep dental care or immunodepression. The improvement of this dental unit was necessary by putting a filter of 0.2 microns porosity before the arrival of the water in hand-pieces allowing the limitation of FLA passage.

Biofilms augment the number of free-living amoebae in dental unit waterlines.

Freshwater amoebae are ubiquitous. Some species can cause infections in humans while others can ingest and protect opportunistic bacteria. Although the presence of free-living amoebae in various water sources has been reported, few studies have looked at their concentration, which may be clinically relevant, especially if they are present in healthcare devices. A simple technique was used to detect, observe, and evaluate the concentration of free-living amoebae in dental unit and tap water samples. Fifty-three water samples were collected from 35 dental units (air/water syringes) and 18 water taps. The technique was based on the ability of waterborne bacteria to create a biofilm and serve as substratum for the development of amoebae naturally present in the water samples. Laboratory-grown freshwater biofilms support the proliferation of a wide variety of free-living amoebae. All the dental unit water samples tested contained amoebae at concentrations up to 330/mL, or more than 300 times the concentration in tap water from the same source. Hartmanella, Vanella, and Vahlkampfia spp. were the most frequently encountered. Naegleria and Acanthamoeba spp. were also present in 40% of the samples. Four of the samples collected from dental units, but none from water taps, contained amoebae able to proliferate at 44 degrees C. Biofilms that form inside some dental instruments can considerably increase the concentration of free-living amoebae, some of which are potential human pathogens.