Overlevelse af indikatororganismer og patogener i ledningsnet

Following several of the larger contamination incidents in drinking water supplies, indicator organisms (coliforms, E. coli, enterococci) have been present in the distribution network for a long time even after extensive flushing. For example E. coli and coliforms were present in the distribution system of Aarhus for 5 to 6 weeks (2002), without the source being identified.

Numerous observations regarding the presence of indicator organisms in drinking water distribution systems can not be explained by the current knowledge. In order to evaluate the significance of such presence of coliforms, it is necessary to gain further knowledge on which circumstances they can survive - or maybe even grow - in distribution systems.

The purpose of this project was to investigate the survival of selected indicator organisms and pathogens in contact with different types of pipe material and in pipes collected from distribution systems with special focus on the interaction between water phase and biofilm. Furthermore the purpose was, to investigate the influence of working procedures when replacing distribution pipes, including whether renovation of pipes represents a risk for the microbial drinking water quality.

Firstly, the working procedures during renovation work were mapped. An assessment of the working procedures at Nordvand A/S identified a series of risks for the microbial drinking water quality before, during and after the renovation work. Based on the identified risks a number of recommendations for the water supplies were prepared – including storage of new pipe preventing the inner surfaces to get in contact with the environment, the renovation work should be performed as controlled as possible, the staff should be trained in hygiene, defined procedures for the renovation work etc.

Microbial analysis of the water from renovation dig-outs revealed high bacterial counts and a significant level of indicator organisms (coliforms, E. coli, enterococci). This water can cause problems in the distribution network if it gets in contact with the inner surfaces of the new water pipes. Therefore the working procedure during renovation should always include removal of water from the digouts.

This project has lead to Nordvand A/S having more structured work procedures for pipe renovation. After the implementation of the new procedures, Nordvand A/S has only experienced few occurrences of high bacterial counts in relation to pipe renovation. Thus the project demonstrated that high bacterial counts following pipe renovation can be prevented by good hygiene and standardised working procedures.

To evaluate the risk of a microbial contamination in relation with renovation work, the survival of a number of bacteria was investigated. This work included seven strains of Escherichia coli, seven strains of Klebsiella pneumoniae and six strains of Campylobacter jejuni, isolated from both clinical patients and the environment.

Initial concentrations of E. coli and K. pneumoniae were 100-200 cells/ml and of C. jejuni 1000 celler/ml, which are low concentrations close to realistic levels. All strains of the indicators E. coli and K. pneumoniae survived for more than four weeks in drinking water, which is a relative long time considering that the residence time of drinking water in distribution networks usually is shorter than one week.

Strains of the pathogen C. jejuni were only detected in the water phase for two to five days by the applied analysis method. In general strains of K. pneumoniae (T½: 11-42 days) survived better than strains of E. coli (T½: 6-18 days). All strains of K. pneumoniae and E. coli could also be detected in the biofilm on PE-pipes, while none of the C. jejuni could be detected in biofilms.

Generally K. pneumoniae colonised the biofilm of the pipe surfaces to a higher degree than E. coli. However, the water phase concentration of K. pneumoniae and E. coli was always higher than the biofilm concentration. K. pneumoniae and E. coli always decayed significantly in the water phase during the four weeks investigated, while the biofilm concentration was relatively stable.

This indicated a better survival in the biofilm. The survival of E. coli and K. pneumonia in drinking water was inhibited by the presence of plastic materials (PE (polyethylene), PEX (cross-linked polyethylene) and silicone) and the degree of the effect depended on material type. Silicone had the strongest inhibition on the survival in the water phase followed by PE and PEX (silicone >> PE > PEX).

Both indicator organisms were detected in the biofilm on all three materials during the four weeks investigated. Contact with plastic pipes had no effect on the survival of C. jejuni in drinking water and C. jejuni was not detected in biofilms on any of the materials. There was no clear distinction between the colonisation of the biofilm on the three materials, though the biofilm density was higher on silicone than on PE and PEX.

It was investigated how indicator organisms and pathogens survived under realistic conditions, i.e. in pipes collected from a real distribution system, with scalings and mature biofilms. Both indicator organisms - E. coli and K. pneumoniae – were detected both in the water phase and in the biofilm for more than four weeks in PE pipes, but only for two weeks in cast iron pipes.

C. jejuni was detected in the pipes for up to three days by the applied analysis method, and only in the water phase. C. jejuni might be present in a viable-but-not-culturable state, but it is unknown whether the cells are infectious in this state. There were significantly more bacteria (total ATP) in the water phase of cast iron pipes than in the investigated PE pipes, and the biofilm density on cast iron pipes was larger than on PE pipes.

The older the PE pipe was the higher was the biofilm density and the higher degree of colonisation of the biofilm with indicator organisms. The project has demonstrated that high bacterial counts following pipe renovation can be prevented by good hygiene and standardised working procedures. It is important that water from the renovation dig-outs do not get in contact with the inner surfaces of the new water pipes, since this can affect the microbial drinking water quality.

Laboratory investigations with new and old pipes collected from real distribution systems demonstrated that indicator organisms can colonise the surfaces of pipes and survive for more than four weeks. Laboratory investigations demonstrated that pipe material influenced the survival of indicator organisms and most likely also the survival of pathogens, with indicator organisms surviving in drinking water for two weeks in cast iron pipes and four weeks in plastic pipes (PE, PEX).