Successful isolation and cultivation of Cylindrospermopsis raciborskii strains isolated from finished drinking water samples

Menezes, Carina; Valério, Elisabete; Botelho, Maria João; Martins, Olga; Dias, Elsa

http://hdl.handle.net/10400.18/6707

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This work presents the successful establishment of Cylindrospermosis raciborskii cultures isolated from water samples collected at the exit point of Water Treatment Plant (WTP). An intense bloom dominated by filamentous cyanobacteria (Aphanizomenon spp., Planktothrix spp., Cylindrospermosis raciborskii, Anabaena spp.) occurred in the summer of 2015 in Roxo reservoir (south Portugal). Several cyanotoxins (microcystins, saxitoxins, cylindrospermopsin) were detected in raw and treated water, although at levels below the corresponding regulatory and/or guideline values. Nevertheless, this bloom caused intense unpleasant odour and taste in the water supplied to the populations and cyanobacterial cells (up to 1000 cells.mL-1) were detected in finished water samples collected at the exit point of WTP. Treated water samples collected at the WTP and at the city water deposit were inoculated in Z8 culture medium and cyanobacterial growth was followed by optical microscopy. After 30 days, cyanobacterial growth was observed showing resistance to the treatment processes and maintenance of reprodution capacity. Interestingly, morphometric and molecular analysis revealed the presence of C. raciborskii. Three isolates of this species were obtained and none were cylindrospermopsin- or microcystins-producers, as confirmed by Enzyme Linked Immunosorbent Assays (ELISA) and by amplification of genes (PS, PKS, mcyA-cd, mcyAB, mcyB) involved in those cyanotoxin synthesis. However, the ELISA for saxitoxins was positive for the 3 isolates and confirmation of this toxin production is in progress. To our knowledge, this is the first report on the establishment of successful cultures of C. raciborskii that survived to conventional water treatment processes.