Determination of minimal concentration of Piscirickettsia salmonis in water columns to establish a fallowing period in salmon farms.

Abstract A highly sensitive real-time PCR procedure to detect and quantify the number of Pisciricketsia salmonis units in seawater samples from affected farm sites has been developed. The purpose was to determine a fallowing period that would allow safe restocking of the target farm with new fish. Bacterial load was determined in water samples by comparing the obtained amplification values against a standard curve generated by the amplification of known concentrations of the ITS-ribosomal component of P. salmonis DNA, cloned in a suitable vector. The standard curve was linear over the range of 10(1)-10(10) log units. Target samples were taken every 10 days over a 40-day period, at 5 m depth and at the surface. In a highly affected area of southern Chile, the number of bacterial units in farm water decreased to zero at day 50. Therefore, a fallowing period of 50 days post-removal of cages of affected fish appears to be appropriate before restocking. This procedure could be adapted to control disease problems because of other pathogens in fish farm waters.