Comparison of two molecular methods used for subtyping of Legionella pneumophila 1 strains isolated from a hospital water supply.

The results of the pulsed-field gel electrophoresis and the sequence-based typing (using the loci flaA, pilE, asd, mip, mompS and proA) were compared for subtyping of Legionella pneumophila 1 strains isolated from a hospital water supply. Molecular typing was carried out on 61 isolates (38% of the positive samples) selected on space and temporal criteria in order to follow the evolution of the water-system colonization. For all the 61 isolates, the sequence of the amplified mip gene fragment identified Legionella pneumophila strain Wadsworth. Genotype testing by PFGE analysis showed three different patterns, correspondent to three SBT types according to the allelic profiles. Both PFGE and SBT indicated the circulation and the persistence in the hospital potable water-system of three types randomly distributed in space and time. The two molecular methods adopted showed a 100% concordance, although a low degree of genetic heterogeneity characterized the isolates. The electrophoretic patterns were sufficiently unambiguous to consider PFGE a highly discriminatory typing method, but the SBT technique besides accurately characterizing isolates, was able to identify Legionella strains through analysis of the mip gene. A typing method with this level of discriminatory power has great potential for assisting in epidemiological studies.

Molecular epidemiology of Legionella pneumophila serogroup 1 isolates following long-term chlorine dioxide treatment in a university hospital water system.

This paper describes the results of a five-year monitoring programme applied to the water distribution system of the University Hospital of Pisa (Italy). The purpose of the programme was to evaluate the efficacy of an integrated water safety plan in controlling Legionella spp. colonisation of the potable water system. The impact of the safety plan on the ecology of legionella in the water network was evaluated by studying the genetic variability and the chlorine susceptibility of the strains isolated prior to, and throughout, the application of continuous chlorine dioxide treatment. After 45 months of water hyperchlorination, Legionella spp. were still present but the positive supply points were reduced by 79.4%. The samples exceeding 10(3)cfu/L were reduced by 83.8% and the mean counts showed a decrease of 94.6%. The majority of the isolates belonged to Legionella pneumophila serogroup 1 (overall positivity rate: 161/423; 38%). Molecular typing was performed on 61 isolates (37.9% of the positive samples) selected on spatial and temporal criteria. This revealed the circulation and the persistence in the hospital environment of three prevalent types of L. pneumophila Wadsworth, demonstrating allelic and electrophoretic characteristic profiles and different chlorine susceptibility. Two of these, one predominant and pre-dating the sanitation regimen, and one other isolated after three years of water treatment, were chlorine tolerant. Despite the ineffectiveness of chlorine dioxide in eradicating L. pneumophila, the risk management plan adopted appeared to discourage further cases of nosocomial legionellosis.