Dataset Information

Cultural and Molecular Evidence of Legionella spp. Colonization in Dental Unit Waterlines: Which Is the Best Method for Risk Assessment?

ABSTRACT: Legionella spp. are ubiquitous in aquatic habitats and water distribution systems, including dental unit waterlines (DUWLs). The aim of the present study was to determine the prevalence of Legionella in DUWLs and tap water samples using PMA-qPCR and standard culture methods. The total viable counts (TVCs) of aerobic heterotrophic bacteria in the samples were also determined. Legionella spp. were detected and quantified using the modified ISO 11731 culture method. Extracted genomic DNA was analysed using the iQ-Check Quanti Legionella spp. kit, and the TVCs were determined according to the ISO protocol 6222. Legionella spp. were detected in 100% of the samples using the PMA-qPCR method, whereas these bacteria were detected in only 7% of the samples using the culture method. The number of colony forming units (CFUs) of the TVCs in the DUWL and tap water samples differed, with the bacterial load being significantly lower in the tap water samples (p-value = 0). The counts obtained were within the Italian standard range established for potable water in only 5% of the DUWL water samples and in 77% of the tap water samples. Our results show that the level of Legionella spp. contamination determined using the culture method does not reflect the true scale of the problem, and consequently we recommend testing for the presence of aerobic heterotrophic bacteria based on the assumption that Legionella spp. are components of biofilms.

SUBMITTER: Ditommaso S

PROVIDER: S-EPMC4772231 | biostudies-other | 2016 Feb

REPOSITORIES: biostudies-other

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Cultural and Molecular Evidence of Legionella spp. Colonization in Dental Unit Waterlines: Which Is the Best Method for Risk Assessment?

Ditommaso Savina S Giacomuzzi Monica M Ricciardi Elisa E Zotti Carla M CM

International journal of environmental research and public health 20160206 2

Legionella spp. are ubiquitous in aquatic habitats and water distribution systems, including dental unit waterlines (DUWLs). The aim of the present study was to determine the prevalence of Legionella in DUWLs and tap water samples using PMA-qPCR and standard culture methods. The total viable counts (TVCs) of aerobic heterotrophic bacteria in the samples were also determined. Legionella spp. were detected and quantified using the modified ISO 11731 culture method. Extracted genomic DNA was analys ...[more]

PMID: 26861373

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Project description:BackgroundDental unit waterlines (DUWLs) provide water for handpieces, air/water syringes, and mouth-rinse water outlets. DUWL contamination can negatively affect the operating environment and public health. Therefore, it is important to elucidate the bacterial concentrations and microbial composition in the DUWLs from different dental specialties.MethodsWe collected 350 5-mL dental water samples (from high-speed handpieces, air/water syringes, and mouth-rinse water outlets) from 60 dental chair units (DCUs) at a dental hospital to determine the bacterial concentrations by culture methods. Meanwhile, to investigate the diversity and community structure of microbe in the DUWLs, 17 high-quality DNA from 60 250-mL air/water syringe water samples, which were collected from the same 60 DCUs, were analyzed using 16S rDNA high-throughput sequencing.ResultsThe median bacterial concentration was 166 (31.5, 672.5) CFU/mL and the range was 0-3,816,000 CFU/mL. Only 42.6% of the water samples had bacterial concentrations below 100 CFU/mL. The Kruskal-Wallis H-test revealed that the water samples from three dental specialties had significantly different bacterial concentrations (H = 27.441, P < 0.01). High-throughput sequencing results showed significant differences in bacterial community structure between periodontics and the other two dental specialties. In the samples from three dental specialties, 508 OTUs were detected, with 160, 182 and 176 OTUs unique to the periodontics, endodontics and prosthodontics specialties, respectively. Linear discriminant analysis (LDA) effect size (LEfSe) suggested that Hydrocarboniphaga, Zoogloea, Aquabacterium, and Hydrogenophaga were enriched in the periodontics specialty; Acinetobacter, Geothrix, and Desulfovibrio were enriched in the prosthodontics specialty; and Alistipes, Clostridium XIVa, and Serratia were enriched in the endodontics specialty. Seven potentially human-pathogenic genera (Pseudomonas, Acinetobacter, Sphingomonas, Ochrobactrum, Rhizobium, Brevundimonas, and Methylobacterium) with relative abundance exceeding 1% were also detected in the DUWLs.ConclusionsThe bacterial concentrations and microbial composition were influenced by different dental specialties, so a validated disinfection protocol should be used to control DUWL contamination in different dental specialties.

Project description:Dental unit water lines (DUWL) are susceptible to biofilm development and bacterial growth leading to water contamination, causing health and ecological effects. This study monitors the interactions between a commonly used nanosilver disinfectant (ASAP-AGX-32, an antimicrobial cleaner for dental units, 0.0032% Ag) and biofilm development in DUWL. To simulate the disinfection scenario, an in-house DUWL model was assembled and biofilm accumulation was allowed. Subsequent to biofilm development, the disinfection process was performed according to the manufacturer's instructions. The pristine nanosilver particles in the cleaner measured between 3 and 5 nm in diameter and were surrounded by a stabilizing polymer. However, the polymeric stabilizing agent diminished over the disinfection process, initiating partial AgNPs aggregation. Furthermore, surface speciation of the pristine AgNPs were identified as primarily AgO, and after the disinfection process, transformations to AgCl were observed. The physicochemical characteristics of AgNPs are known to govern their fate, transport and environmental implications. Hence, knowledge of the AgNPs characteristics after the disinfection process (usage scenario) is of significance. This study demonstrates the adsorption of AgNPs onto biofilm surfaces and, therefore, will assist in illustration of the toxicity mechanisms of AgNPs to bacteria and biofilms. This work can be an initial step in better understanding how AgNPs transform depending on the conditions they are exposed to during their lifetime. Until this date, most research has been focused on assessing the impacts of pristine (lab synthesized) nanomaterials on various systems. However, it is our belief that nanoparticles may undergo transformations during usage, which must be taken into consideration. Furthermore, this experiment is unique as it was conducted with a commonly used, commercially available nanosilver suspension leading to more realistic and applicable findings.

Dataset Information

Cultural and Molecular Evidence of Legionella spp. Colonization in Dental Unit Waterlines: Which Is the Best Method for Risk Assessment?

Publications

Cultural and Molecular Evidence of Legionella spp. Colonization in Dental Unit Waterlines: Which Is the Best Method for Risk Assessment?

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