Malgorzata Tarczynska

Establishment of an Alert Level Framework for cyanobacteria in drinking water resources by using the Algae Online Analyser for monitoring cyanobacterial chlorophyll a.

The Algae Online Analyser (AOA) fluorometer simultaneously distinguishes four different phytoplankton groups by their specific fluorescence spectra and thus allows for real-time in-situ chlorophyll a measurements per algal group. This AOA was used for monitoring cyanobacterial chlorophyll a in the drinking water at the Bronisławow Bay abstraction point in Sulejow Reservoir (Poland). The main goal of this research was to develop an early warning method for the detection of cyanobacterial biovolume in the source water, in order to establish an Alert Level Framework for the drinking water abstraction point in Sulejow Reservoir. A positive correlation between cyanobacterial biovolume, as determined by conventional methods, and cyanobacterial chlorophyll a, as measured by the AOA, was found (p<0.05). The results of this study were used to determine threshold values for the Alert Level Framework, based on cyanobacterial chlorophyll a concentrations in the source water of Sulejow Reservoir. The presented threshold values are determined specifically for this abstraction point, but the principles can be applied to other locations.

Effect of microcystin-LR and cyanobacterial extract from Polish reservoir of drinking water on cell cycle progression, mitotic spindle, and apoptosis in CHO-K1 cells.

Microcystin-LR is a cyanobacterial toxin possessing a potent tumor-promoting activity mediated through inhibition of protein phosphatases PP1 and PP2A. Because these enzymes are involved in fundamental cell processes, we decided to examine the influence of microcystin-LR on cell cycle progression, onset of anaphase, segregation of chromosomes by the mitotic spindle, and apoptosis in Chinese hamster ovary (CHO-K1) cells. Cells were incubated with 25, 50, and 100 microM of pure microcystin-LR and a cyanobacterial extract for 14, 18, and 22 h. Giemsa staining of cells treated with these toxins revealed a dose- and time-dependent increase of mitotic indices, accumulation of abnormal G(2)/M figures with hypercondensed chromosomes, abnormal anaphases with defective chromosome separation, and polyploid cells. Because spindle checkpoint is a fundamental regulatory mechanism that assures the onset of anaphase and subsequent exit from mitosis, we examined the spindle organization in microcystin-treated cells. The majority of the mitotic cells showed monopolar and multipolar mitotic spindles (multiple asters). Microtubule bundles were present in interphase cells. Our results indicate that microcystin-LR induces apoptosis and necrosis in a dose- and time-dependent manner and that the frequency of dead cells cells is positively correlated with the frequency of polyploid cells.

The toxicity of cyanobacterial blooms as determined by microbiotests and mouse assays

Cyanobactrial blooms are common throughout the world in eutrophic water bodies. Cyanobacterial hepato- and neurotoxins represent a health threat for humans as well as for animals in drinking water supplies. Mouse bioassays have been used extensively to detect cyanobacterial blooms toxicity but the use of warm-blooded vertebrates is limited to specialised laboratories and poses ethical problems. Hence, it is necessary to find alternative assays to detect and quantify cyanobacterial hazards. This study reports on the toxicity of 7 cyanobacterial bloom samples from the Jeziorsko. Reservoir in central Poland as determined with 4 microbiotests: the Thamnotoxkit F™, the Daphtoxkit F™ magna, the ProtoxkitF™ and the Spirotox. The phytoplankton samples were lyophilized, sonicated and passed through octadecyl cartridges. The results were compared to those obtained with mouse bioassays and HPLC analysis of microcystin-LR (MC/LR) in the bloom samples..