The occurrence of the cyanobacterial toxins anatoxin-a (ATX) and cylindrospermopsin (CYN) in surface waters has been reported throughout the world. Beside degradation, sorption is an important pathway for toxin elimination if these resources are used for drinking water production via sediment passage. However, to date studies that systematically investigated sorption of these toxins onto sediments are lacking. Therefore, the aim of our work was (i) to determine the adsorption coefficients of ATX and CYN according to the Freundlich and Langmuir model for sediments of various textures and (ii) to derive sorptionrelevant sediment characteristics. We determined sorption parameters in air-dried samples of eight differently textured sediments using batch experiments. Results for both toxins showed best fits with the Langmuir model. Organic C proved to be the main sediment parameter determining CYN sorption. There was no or little CYN sorption on sandy and silty sediments (0e39 mg kg-1), respectively, presumably due to charge repulsion from the negatively charged surfaces. Sorption of ATX (max. sorbent loading ranging from 47 to 656 mg kg-1) was much stronger than that of CYN (max. sorbent loading ranging from 0 to 361 mg kg-1) and predominantly controlled by clay and to a minor degree also by organic C and silt. While ATX sorption to most sediments occurred mainly through cation exchange this mechanism played only a minor role in CYN sorption to organic C. Hence, high mobility for CYN and moderate mobility for ATX during sediment passage has to be expected.

Cyanobacteria of the order Nostocales – native species as well as alien species from tropical regions – were found to increase in many Brandenburg lakes while the formerly dominating microcystin (MC) producing cyanobacteria (Microcystis and Planktothrix) occurred less often and in lower amounts. As a consequence, lower MC concentrations were observed while the toxin cylindrospermopsin (CYN) that is produced by Nostocales was found to be widely distributed and to exceed sometimes the recommended guideline value for drinking-water of 1 µg L-1. Recent data on the occurrence of further neurotoxins (paralytic shellfish poisoningtoxin, PSP and anatoxin, ATX) produced by cyanobacteria of the order Nostocales did not exist. Nostocales are superior competitors under conditions of high light intensity and nitrogen depletion because they can fix molecular nitrogen. Their germination is regulated by temperature and the temporal starting point of the pelagic population determines the population size (the earlier the larger). Therefore, the following working hypothesis has been put forward: Combined effects of declining trophic state and global warming favor the development of Nostocales and cause a shift in the species composition as well as in the occurrences of cyanobacterial toxins. The NOSTOTOX project aimed to determine the present occurrence and future development of Nostocales and their toxins in waterbodies. Special emphasis was paid to answer the question, which Nostocales species and which toxins can be expected under conditions of a proceeding decline in trophic state and increasing water temperature. The outcome of the project aims to contribute to developing recommendations and guidelines for the management of inland waters and drinking water supplies.

Recent results show that cylindrospermopsin is more frequent and widespread in surface waters than previously assumed. Studies on the fate of CYN in sediments are lacking, but this is important if these resources are used for drinking-water production via sediment passage. Therefore, the aim of our study was to determine a) CYN retention in two sandy sediments as a function of flow rate, CYN concentration, the presence of DOM and the content of fines (1% and 4%, respectively) and b) the influence of sediment preconditioning and DOM composition of the water (aquatic DOM versus DOM released from lysed cells) on CYN degradation. Retention of CYN proved negligible under the investigated conditions. Degradation in virgin sediments showed the highest lag phases (20 days). Preconditioned sediments showed no lag phase. The presence of aquatic DOM yielded highest degradation rates (k1 ¼0.46 and 0.49 day 1) without a lag phase. Readily available organic carbon sources were preferentially metabolized and hence induced a lag phase. Thus, the presence and composition of DOM in the water proved important for both CYN degradation rates in preconditioned sediments and for the lag phase. Cylindrospermopsin degradation took place solely in the sediment and not in the water body.

Some tropical cyanobacteria have spread to temperate freshwaters during the last decades. To evaluate their further development in temperate lakes, we studied the temperature- and light-dependent growth of three invasive (Cylindrospermopsis raciborskii, Anabaena bergii and Aphanizomenon aphanizomenoides) and three native (Aphanizomenon gracile, Aphanizomenon flos-aquae and Anabaena macrospora) cyanobacterial species (Nostocales) from German lakes. We also included one potentially invasive (Aphanizomenon ovalisporum) Nostocales species. We conducted semi-continuous culture experiments and a microcosm experiment along a natural light gradient. Temperature data were used to design a model to simulate the development of selected species according to three temperature scenarios (past, present and future). Native species had significantly higher growth rates than invasive species and the potential invader A. ovalisporum at low temperatures (<= 10 °C), while the opposite was true at high temperatures (>= 35 °C). Maximum growth rates of A. ovalisporum, A. aphanizomenoides and C. raciborskii were clearly higher than those of A. bergii and the native species. Regarding light-dependent growth, significant differences were found between single species but not between all native and invasive species. The model simulation demonstrates a shift in dominance from the native A. gracile in the historic scenario to C. raciborskii populations in the future scenario, in which also the potential invader A. ovalisporum is able to establish populations in temperate lakes. Our findings suggest that any further temperature increase would promote the growth and development of Nostocales species in general, and that of the invasive species in particular, and would enable a more northward expansion of A. ovalisporum.

The neurotoxin anatoxin-a (ATX), has been detected in several northeast German lakes during the last two decades, but no ATX producers have been identified in German water bodies so far. In 2007 and 2008, we analyzed phytoplankton composition and ATX concentration in Lake tolpsee (NE Germany) in order to identify ATX producers. Sixty-one Aphanizomenon spp. strains were isolated, morphologically and phylogenetically characterized, and tested for ATX production potential by liquid chromatography–tandem mass spectrometry (LC–MS/MS). New primers were specifically designed to identify a fragment of a polyketide synthase gene putatively involved in ATX synthesis and tested on all 61 Aphanizomenon spp. strains from L. Stolpsee and 92 non-ATX-producing Aphanizomenon spp., Anabaena spp. and Anabaenopsis spp. strains from German lakes Langersee, Melangsee and Scharmützelsee. As demonstrated by LC–MS/MS, ATX concentrations in L. Stolpsee were undetectable in 2007 and ranged from 0.01 to 0.12 µg l-1 in 2008. Fifty-nine of the 61 strains isolated were classified as Aphanizomenon gracile and two as Aphanizomenon issatschenkoi. One A. issatschenkoi strain was found to produce ATX at concentrations of 2354 ± 273 µg g-1 fresh weight, whereas the other A. issatschenkoi strain and A. gracile strains tested negative. The polyketide synthase gene putatively involved in ATX biosynthesis was found in the ATX-producing A. issatschenkoi strain from L. Stolpsee but not in the non-ATX-producing Aphanizomenon spp., Anabaena spp. and Anabaenopsis spp. strains from lakes Stolpsee, Langersee, Melangsee, and Scharmützelsee. This study is the first confirming A. issatschenkoi as an ATX producer in German water bodies.

Neurotoxic paralytic shellfish poisoning (PSP) toxins, anatoxin-a (ATX), and hepatotoxic cylindrospermopsin (CYN) have been detected in several lakes in northeast Germany during the last 2 decades. They are produced worldwide by members of the nostocalean genera Anabaena, Cylindrospermopsis, and Aphanizomenon. Although no additional sources of PSP toxins and ATX have been identified in German water bodies to date, the observed CYN concentrations cannot be produced solely by Aphanizomenon flos-aquae, the only known CYN producer in Germany. Therefore, we attempted to identify PSP toxin, ATX, and CYN producers by isolating and characterizing 92 Anabaena, Aphanizomenon, and Anabaenopsis strains from five lakes in northeast Germany. In a polyphasic approach, all strains were morphologically and phylogenetically classified and then tested for PSP toxins, ATX, and CYN by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and enzyme-linked immunosorbent assay (ELISA) and screened for the presence of PSP toxin- and CYN-encoding gene fragments. As demonstrated by ELISA and LC-MS, 14 Aphanizomenon gracile strains from Lakes Melang and Scharmützel produced four PSP toxin variants (gonyautoxin 5 [GTX5], decarbamoylsaxitoxin [dcSTX], saxitoxin [STX], and neosaxitoxin [NEO]). GTX5 was the most prevalent PSP toxin variant among the seven strains from Lake Scharmützel, and NEO was the most prevalent among the seven strains from Lake Melang. The sxtA gene, which is part of the saxitoxin gene cluster, was found in the 14 PSP toxin-producing A. gracile strains and in 11 non-PSP toxin-producing Aphanizomenon issatschenkoi, A. flos-aquae, Anabaena planktonica, and Anabaenopsis elenkinii strains. ATX and CYN were not detected in any of the isolated strains. This study is the first confirming the role of A. gracile as a PSP toxin producer in German water bodies.

In summer 2007 & 2008, 100 water samples were collected from 10 freshwater reservoirs with cyanobacteria issues. Phytoplankton was determined according to the Utermohl method [1]. Intra- and extracellular CYN, ATX-a, STX were analyzed by LC-MS-MS or HPLC-PDA at UBA, and in addition, Veolia tested Abraxis ELISA kits for total CYN and total STX on the 2008 water samples (n=45). Cyanobacterial abundance was comparably low in 2007 & 2008 for all reservoirs, probably because of cooler summer months, with less sunlight, more rain and quickly decreasing fall temperatures (except in reservoir 10, which had low incoming nutrient charges). For instance, average chlorophyll content was 12 µg/L in 2007 and 35 µg/L in 2008 in Western France, when 60-80 µg/L concentrations are usually measured. In spite of these environmental conditions, cyanobacteria were detected in 97% of the samples and cyanotoxins in 55%. WHO level 3 for drinking water (>100 000 cell/mL) was reached for 20-25% of the samples. Among the species observed in the water samples, the following potential CYN, ATX-a, STX producers were observed: Cyanotoxin LC-MS-MS and HPLC-PDA results are given on the right. ELISA results for CYN and STX of the 2008 samples only partially agree with the LC-MS-MS data. This might be due to the differences in extraction procedures of the two methods, cross-reactivity issues of the ELISAs for derivatives, in combination with overall very low concentrations of the toxins.