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BMC Microbiol. 2013 Feb 26;13:49. doi: 10.1186/1471-2180-13-49.

Detection limits of Legionella pneumophila in environmental samples after co-culture with Acanthamoeba polyphaga.

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Swiss National Reference Centre for Legionella, Cantonal Institute of Microbiology, Via Mirasole 22a, 6500, Bellinzona, Switzerland.



The efficiency of recovery and the detection limit of Legionella after co-culture with Acanthamoeba polyphaga are not known and so far no investigations have been carried out to determine the efficiency of the recovery of Legionella spp. by co-culture and compare it with that of conventional culturing methods. This study aimed to assess the detection limits of co-culture compared to culture for Legionella pneumophila in compost and air samples. Compost and air samples were spiked with known concentrations of L. pneumophila. Direct culturing and co-culture with amoebae were used in parallel to isolate L. pneumophila and recovery standard curves for both methods were produced for each sample.


The co-culture proved to be more sensitive than the reference method, detecting 10²-10³ L. pneumophila cells in 1 g of spiked compost or 1 m³ of spiked air, as compared to 10⁵-10⁶ cells in 1 g of spiked compost and 1 m³ of spiked air.


Co-culture with amoebae is a useful, sensitive and reliable technique to enrich L. pneumophila in environmental samples that contain only low amounts of bacterial cells.

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