Identification of Cylindrospermopsin and Cylindrospermopsis raciborskii from Anuradhapura District, Sri Lanka

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Cylindrospermopsin (CYN) is one of the potent cyanotoxin which affects the functions of kidney and liver. This study was performed to identify CYN in Anuradhapura water reservoirs using molecular and biochemical methods. Water samples were collected from Kala wewa, Nuwara wewa, Tissa wewa and Jaya ganga. Under microscope, Cylindrospermopsis species were recorded as the dominant cyanobacterial species along with Microcystis, Anabaena, Chroococcus, Phormidium, Oscillatoria, which were comparatively moderate to low. In molecular detection, the presence of cyanobacteria, the presence of Cylindrospermopsis and Cylindrospermopsis strains that have the genetic potential to produce CYN were detected using specific PCR primers targeting 16S rRNA gene, C. raciborskii specific cylindrospermopsin synthetase gene and cylindrospermopsin specific peptide synthase (PS) gene respectively. The presence of toxin producing Cylindrospermopsis raciborskii in those water samples was reconfirmed by a nested PCR using C. raciborskii specific primers cyl2, cyl4 and cyl-int and by direct sequencing. Further, ELISA detection kit confirmed the presence of CYN and the toxin concentration. All were positive for CYN with mean concentrations of 0.104, 0.091, 0.255 and 0.0967 ng/ml for Jaya ganga, Kala wewa, Nuwara wewa and Tissa wewa respectively. Water samples collected from water purification centre in Anuradhapura showed 0.245 and 0.154 ng/ml of CYN before purification and after addition of chlorine respectively.