Detection of legionella with an amoebal co-culture method

Abstract

On the authority of the Netherlands VROM-Inspectorate, the RIVM has developed a new method for detecting live Legionella bacteria in water. The method known as the amoebal co-culture method can be used in addition to the regular culture method using agar plates, which probably does not detect all harmful Legionella bacteria in water samples. The co-culture method is particularly suitable for samples that contain a lot of other bacteria, like samples from sewage treatment plants.

With the regular culture methods, water samples are applied to agar plates that contain specific nutrients on which Legionella can grow and be seen as colonies. Other, non-Legionella, bacteria can also grow on the agar plates and inhibit the growth of Legionella so that they are hard to detect. Moreover, Legionella bacteria at a certain stage (viable-but-not-culturable) cannot grow on agar plates.

With the amoebal co-culture method, water samples are placed on amoebae. These single-celled organisms are the natural host for Legionella, in which the bacteria can multiply. This has the advantage of stimulating the growth of Legionella and not other bacteria. This process of enrichment allows Legionella bacteria to subsequently be detected by plating on agar plates.

In the Netherlands, in-premise plumbing and cooling towers are checked regularly for Legionella in order to prevent human exposure to Legionella. A Legionella infection can lead to a severe form of pneumonia. When people have incurred a Legionella infection, it is important to track down and eradicate the source of the bacteria to prevent further exposure. Legionella is detected with the regular culture method on agar plates. The amoebal co-culture method can now also be used for detecting the bacteria, especially for samples where the bacteria is difficult to detect.