Der Großteil von Seen und Flüssen in Deutschland befindet sich nicht in dem von der europäischen Wasserrahmenrichtlinie geforderten guten ökologischen Zustand. Die Ursache hierfür besteht in den meisten Gewässern nach wie vor in zu hohen Nährstoffbelastungen. Dadurch wird besonders im Sommer das Wachstum des Phytoplanktons (Algen) gefördert, das Wasser wird trübe, zeitweise sauerstoffarm und riecht unangenehm. Solche Gewässer stellen für viele Tiere und Pflanzen keinen geeigneten Lebensraum dar und sind für den Menschen unattraktiv. Die Hauptnährstoffe, um die es dabei geht, sind Stickstoff und Phosphor. Dabei galt Phosphor (P) lange Zeit als der begrenzende Faktor der Phytoplanktonbiomasse in Binnengewässern: Je geringer die PKonzentration desto geringer die Biomasse und desto besser die Gewässergüte. Dies ist bis heute Lehrbuchmeinung. In der Praxis wurde und wird daher auf eine Senkung der Phosphorkonzentrationen gesetzt, was in vielen, aber längst nicht allen Gewässern zum Erfolg führte. Deutlich weniger Studien zeigten, dass Stickstoff die Phytoplanktonbiomasse begrenzt, was allerdings auch darauf zurückzuführen ist, dass deutlich weniger Studien zum Einfluss von Stickstoff durchgeführt wurden. Eine systematische Analyse zur Bedeutung von Phosphor im Vergleich zu Stickstoff fehlte bisher. Bis heute wird daher die Bedeutung von Stickstoff als begrenzender Faktor der Phytoplanktonbiomasse weitgehend negiert. In NITROLIMIT I (2011 - 2013) wurde dagegen gezeigt, dass die Algenbiomasse in fast der Hälfte der Seen der Norddeutschen Tiefebene durch N begrenzt wird. In der Praxis wird bislang die gezielte Reduktion von Stickstoffeinträgen abgelehnt, weil man befürchtet, dass dies besonders in Seen durch Stickstofffixierung von Cyanobakterien ausgeglichen werden kann und sinkende Nitratkonzentrationen die Freisetzung von Phosphor aus den Gewässersedimenten steigern. Beides könnte einer Verbesserung der Gewässergüte entgegenwirken. Für diese Argumente fehlte jedoch eine fundierte wissenschaftliche Grundlage. Stickstoff wird sowohl in Seen als auch in Fließgewässern intensiv umgesetzt und kann über verschiedene mikrobielle Umsatzprozesse (insbesondere Denitrifikation) auch wieder aus dem System entfernt werden. Fließgewässer transportieren schließlich die nicht zurückgehaltenen Nährstoffe aus den Einzugsgebieten in die Ästuare, Küstengewässer und Meere, wo in weiten Bereichen Stickstoff der limitierende Nährstoff ist. Über den Umsatz und den Rückhalt von Stickstoff in großen Flüssen bestehen bis heute allerdings große Unsicherheiten. Zur Beantwortung der Frage „Ist Stickstoffreduktion ökologisch sinnvoll?“ bestand daher umfangreicher Forschungsbedarf. Die bisherige Strategie zur Verbesserung der Gewässergüte zielte auf Minderung der Phosphorkonzentration ab. Hierzu existieren Erfahrungswerte zu Wirkung und Kosten von Maßnahmen. Viele Maßnahmen zur Phosphorreduktion gehen zu einem gewissen Teil auch mit Stickstoffreduktion einher. Der Erfolg der Begleiterscheinung „Stickstoffreduktion“ wurde jedoch meist nicht analysiert. Fallstudien zur Verbesserung der Gewässergüte durch gezielte Stickstoffminderung wurden bisher nicht durchgeführt, weshalb Daten und Erfahrungen zu Kosten und Wirksamkeit solcher Maßnahmen fehlen. Unabhängig davon, ob eine Verbesserung der Gewässergüte über Phosphor- oder Stickstoffreduktion angestrebt wurde, fehlte bisher eine Strategie, nach der im Voraus Kosten, Wirksamkeit und Nutzen ermittelt und abgewogen werden. Daher bestand auch zur Beantwortung der Frage, „Ist Stickstoffreduktion wirtschaftlich vertretbar?“ deutlicher Forschungsbedarf.

Der Großteil der bundesdeutschen Binnengewässer wird bis 2015 nicht den guten ökologischen Zustand erreichen, der von der EU-Wasserrahmenrichtlinie gefordert wird. Bisher ging man davon aus, dass die Gewässergüte in erster Linie durch Phosphor bestimmt wird. In jüngster Zeit mehrten sich aber Hinweise, dass in vielen Gewässern auch Stickstoff eine entscheidende Steuergröße der Phytoplanktonentwicklung darstellt. Daher wird die Reduzierung von Stickstoffeinträgen gefordert. Die Kosten für Maßnahmen zur Reduktion der Stickstoffeinträge aus punktuellen (beispielsweise Kläranlagen) und diffusen Quellen (beispielsweise aus der Landwirtschaft) werden um ein Vielfaches höher geschätzt im Vergleich zu Maßnahmen zur Reduktion von Phosphoreinträgen. Ob Maßnahmen zur Stickstoffreduktion ökologisch wirksam werden, kann aufgrund unzureichender Kenntnisse zur Herkunft, Umsetzung und Wirkung von Stickstoff derzeit nicht eingeschätzt werden. Daher fordern öffentliche und wirtschaftliche Maßnahmenträger nachdrücklich eine Klärung des Nutzens von Stickstoffelimination. An diesem Punkt setzt NITROLIMIT an. Es sollte eine fundierte wissenschaftliche Grundlage zur Beurteilung des Einflusses von Stickstoff auf die Gewässergüte geschaffen, die Kosten und Nutzen von Maßnahmen zur Verringerung von Stickstoffeinträgen analysiert und darauf basierend Empfehlungen für eine nachhaltige Gewässerbewirtschaftung erarbeitet werden.

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.

Tropical cyanobacteria in German waters: Causes and consequences - Toxic cyanobacteria of tropical origin have spread to water bodies in northern Germany. Here their population size is determined by the onset of germination: the earlier the time of germination, the larger the summer population. Climate-related early increase in water temperatures over the course of the years has promoted their spreading to temperate regions. Toxins known from these species in tropical regions were also found in German lakes. Surprisingly, these toxins are not produced by the invaders but by native species. Thus, the invasion have drawn our attention to an old problem. However, the invaders itself potentially also poses a health risk in German lakes since they produce other – so far unidentified – toxic substances.

The cyanobacterial toxin cylindrospermopsin (CYN) is widely distributed in German lakes, but volumetric data for risk assessment are lacking and it is unclear which cyanobacterial species produce CYN in Europe. We therefore analyzed CYN concentration and cyanobacterial composition of 21 German lakes in 2005. CYN was detected in 19 lakes (102 of 115 samples). In total, 45 samples contained particulate CYN only, and 57 contained both dissolved and particulate CYN. The concentrations were 0.002–0.484 mgL-1 for particulate CYN and 0.08–11.75 mgL-1 for dissolved CYN with a maximum of 12.1 mgL-1 total CYN. A drinking-water guideline value of 1 mgL-1 proposed by Humpage and Falconer [2003. Oral toxicity of the cyanobacterial toxin CYN in male Swiss albino mice: determination of no observed adverse effect level for deriving a drinking water guideline value. Environ. Toxicol. 18, 94–103] was exceeded in 18 samples from eight lakes due to high concentrations of dissolved CYN. CYN occurrence in the German lakes could not be ascribed to the three known CYN-producing species Cylindrospermopsis raciborskii, Anabaena bergii and Aphanizomenon flos-aquae, which were detected in some lakes in low abundances. The highest correlation coefficients were observed between particulate CYN and the native Aphanizomenon gracile. It occurred in 98 CYN-positive samples, was the most abundant Nostocales and was the only Nostocales in five samples. This indicates that A. gracile is a potential CYN producer in German lakes.

Cylindrospermopsis raciborskii, an invasive freshwater cyanobacterium, originated from the tropics but has spread to temperate zones over the last few decades. Its northernmost populations in Europe occur in North German lakes. How such dramatic changes in its biogeography are possible and how its population dynamics in the newly invaded habitats are regulated are still unexplained. We therefore conducted a long-term (1993–2005) study of two German lakes to elucidate the mechanisms behind C. raciborskii population dynamics and to identify the abiotic constraints on its development. Our data revealed that pelagic populations of C. raciborskii thrived for three months during the summer, contributing up to 23% of the total cyanobacteria biovolume. Population sizes varied greatly between years without exhibiting any distinct long-term trends. In the annual lifecycle, C. raciborskii filaments emerged in the pelagic habitat when the temperature rose above 15–17 C. At that time, mean photosynthetically active radiation in the mixed water column (Imix) overstepped its maximum. Rates of population net increase were highest at the beginning of the season (0.15– 0.28 day–1), declined continuously over time, and were significantly positively correlated with Imix. This indicates that the onset of the pelagic population is temperaturemediated and that Imix controls its growth. Since Imix peaks before the population onset, the time of germination is of crucial importance for successful development. To test this hypothesis, we designed a model to simulate pelagic population size, starting at different dates in the annual cycle. Moving the population onset forward by 30 days resulted in a doubling of the population size. We therefore conclude that an earlier rise in water temperature associated with climate change has promoted the spread of C. raciborskii to the temperate zone. Earlier warming permits earlier germination, thereby shifting the pelagic populations to a phase with higher Imix, which advances growth and the population establishment.

Cylindrospermopsis raciborskii, a cyanobacterium of tropical origin, can produce the toxin cylindrospermopsin (CYN). This originally tropical cyanobacterium (blue-green alga) had spread to the waters of the Berlin area. Cylindrospermopsin had been detected in two lakes in the area, but none of the C. raciborskii strains isolated here so far were found to produce the toxin. The main objectives of the CYLIN project were therefore to analyze the distribution and regulation of C. raciborskii and cylindrospermopsin and to determine which cyanobacteria are producing this toxin in order to establish a basis to predict the further course of development of this species and the related health hazards for humans. The CYLIN project was implemented as a three-part program. A screening program was first conducted in 2004 to test regional water bodies for the presence of cylindrospermopsin and potential CYN-producing cyanobacteria in order to obtain an overview of their distribution in the study region. A total of 142 regional water bodies were sampled once each in this qualitative analysis of cylindrospermopsin and cyanobacteria. The screening program was followed by a monitoring program designed to generate quantitative data on the concentrations of dissolved CYN, particulate CYN, cyanobacteria and target environmental parameters at 20 selected lakes, which were sampled 3 times each. Furthermore, we investigated the seasonal dynamics of these parameters at two selected lakes in 2004 and 2005. Apart from this we isolated different cyanobacterial strains and conducted chemical and molecular biological analyses of CYN and CYN-coding genes, in order to identify CYN-producing cyanobacteria. The results show that C. raciborskii and CYN are much more widespread than was previously assumed for the region. C. raciborskii was detected in 22 % of the investigated water bodies, and cylindrospermopsin in 52 %. Additionally, two other toxic cyanobacteria of tropical origin were found for the first time in the Berlin-Brandenburg region, Anabaena bergii and Aphanizomenon aphanizomenoides. The mean and maximum CYN concentrations were 1 µg L-1 and 12 µg L-1, respectively. Since the particulate CYN fraction did not exceed 0.5 µg L-1, the dissolved CYN fraction was found to be responsible for the high CYN concentrations. The proposed guideline value for cylindrospermopsin in drinking water (1 µg L-1; Humpage and Falconer 2003) was exceeded 18 times at 8 different lakes. Although Aphanizomenon flos-aquae (Nostocales) has been unequivocally identified as a producer of cylindrospermopsin, the observed cylindrospermopsin concentrations cannot be attributed to this cyanobacterial species alone. Aphanizomenon gracile was also identified as a potential CYN-producing cyanobacterium. Based on the findings of the CYLIN project, we recommend that cylindrospermopsin be included as in hazard analyisis for drinking and bathing water quality assessments. To identify risk conditions associated with this cyanotoxin, further investigations are needed to identify all cyanobacteria that produce cylindrospermopsin and to elucidate the mechanisms regulating the occurrence of CYN-producing cyanobacteria, CYN synthesis by these organisms, and CYN decomposition in aquatic ecosystems. Our analysis of C. raciborskii population dynamics showed that its germination is temperature-dependent and its population growth light-dependent. Population size was determined by the time of germination, that is, the earlier the time of germination, the bigger the population. Based on these findings, it appears highly likely that the climate-related early rise in water temperatures over the course of the years has promoted the spread of this species to temperate regions. Our hypothesis for the future course of cyanobacterial and cyanotoxin development in German waters is as follows: The combination of trophic decline and global warming works to the general benefit of cyanobacteria of the order Nostocales and leads to a shift in cyanobacterial species and toxin composition. This may ultimately lead to an increase in the incidence of neurotoxins as well as cylindrospermopsin.

Cylindrospermopsis raciborskii, a cyanobacterium of tropical origin, can produce the toxin cylindrospermopsin (CYN). This originally tropical cyanobacterium (bluegreen algae) has now spread to the distant waters of the Berlin area. Cylindrospermopsin has been detected in two lakes in the area, but none of the C. raciborskii strains isolated here so far were found to produce the toxin. The main objectives of the CYLIN project were therefore to analyze the distribution and regulation of C. raciborskii and cylindrospermopsin and to determine which cyanobacteria are producing this toxin in order to establish a basis with which to predict the further course of development of this species and the related health hazards for humans. The CYLIN project was implemented as a three-part program. A screening program was first conducted in 2004 to test regional water bodies for the presence of cylindrospermopsin and potential CYN-producing cyanobacteria in order to obtain an overview of their distribution in the study region. A total of 142 regional water bodies were sampled once each in this qualitative analysis cylindrospermopsin and cyanobacteria. The screening program was followed by a monitoring program designed to generate quantitative data on the concentrations of dissolved CYN, particulate CYN, cyanobacteria and target environmental parameters at 20 selected lakes, which were sampled 3 times each. Last but not least, we investigated the seasonal dynamics of these parameters at two selected lakes in 2004 and 2005. Apart from this we isolated different cyanobacterial strains and conducted chemical and molecular biological analyses of CYN and CYNcoding genes, in order to identify CYN-producing cyanobacteria. The results show that C. raciborskii and CYN are much more widespread than was previously assumed. C. raciborskii was detected in 22 % of the investigated water bodies, and cylindrospermopsin in 52 %. Additionally, two other toxic cyanobacteria of tropical origin were found for the first time in the BerlinBrandenburg region, Anabaena bergii and Aphanizomenon aphanizomenoides. The mean and maximum CYN concentrations were 1 µg L-1 and 12 µg L-1, respectively. Since the particulate CYN fraction did not exceed 0.5 µg L-1, the dissolved CYN fraction was found to be responsible for the high CYN concentrations. The proposed guideline safety value for cylindrospermopsin in drinking water (1 µg L-1) was exceeded 18 times at 8 different lakes. Although Aphanizomenon flos-aquae (Nostocales) has been unequivocally identified as a producer of cylindrospermopsin, the observed cylindrospermopsin concentrations cannot be attributed to this cyanobacterial species alone. Aphanizomenon gracile was also identified as a potential CYN-producing cyanobacterium. Based on the findings of the CYLIN project, we recommend that cylindrospermopsin be included as a risk factor in drinking and bathing water quality assessments. To identify hazard conditions associated with this cyanotoxin, further investigations are needed to identify all cyanobacteria that produce cylindrospermopsin and to elucidate the mechanisms regulating the occurrence of CYN-producing cyanobacteria, CYN synthesis by these organisms, and CYN decomposition in aquatic ecosystems. Our analysis of C. raciborskii population dynamics showed that its germination is temperature-dependent and its population growth light-dependent. Population size was determined by the time of germination, that is, the earlier the time of germination, the bigger the population. Based on these findings, it appears highly likely that the climate-related early rise in water temperatures over the course of the years has promoted the spread of this species to temperate regions. Our hypothesis for the future course of cyanobacterial and cyanotoxin development in German waters is as follows: The combination of trophic decline and global warming works to the general benefit of cyanobacteria of the order Nostocales and leads to a shift in cyanobacterial species and toxin composition. This may ultimately lead to an increase in the incidence of neurotoxins as well as cylindrospermopsin.

The frequent occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) in the (sub)-tropics has been largely associated with cyanobacteria of the order Nostocales of tropical origin, in particular Cylindrospermopsis raciborskii. C. raciborskii is currently observed to spread northwards into temperate climatic zones. In addition, further cyanobacteria of the order Nostocales typically inhabiting water bodies in temperate regions are being identified as CYN-producers. Therefore, data on the distribution of CYN in temperate regions are necessary for a first assessment of potential risks due to CYN in water used for drinking and recreation. A total of 127 lakes situated in the northeastern part of Germany were investigated in 2004 for the presence of the toxin CYN and the phytoplankton composition. The toxin could be detected in half of the lakes (n ¼ 63) and in half of 165 samples (n ¼ 88). Concentrations reached up to 73.2 _g CYN/g DW. CYN thus proved more widely distributed than previously demonstrated. The analyses of phytoplankton data suggest Aphanizomenon sp. and Anabaena sp. as important CYN producers in Germany, and confirm recent findings of Aphanizomenon flos-aquae as CYN-producing species frequently inhabiting water bodies in temperate climatic regions. The data shown here suggest that CYN may be an important cyanobacterial toxin in German water bodies and that further data are needed to assess this.

Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju and Aphanizomenon spp. are both freshwater cyanobacteria of the order Nostocales. C. raciborskii was thought confined to tropical and sub-tropical environments but spread to temperate climatic regions on all continents except Antarctica (Padisák, 1997). It is known to be widely distributed in Northern Germany reaching the northernmost margin of its distribution at latitudes of 53 - 54°N (Krienitz & Hegewald, 1996; Stüken et al., 2006). Strains of this species are detected to produce the hepatotoxin cylindrospermopsin, from which human injury has been clearly identified (Falconer & Humpage, 2006). Nostocales are characterized by trichomal structures such as vegetative cells, heterocysts and akinetes. The heterocysts can fix atmospheric nitrogen (N2) when aquatic nitrogen is depleted (Kim et al., 2005). Akinetes are non-motile, resistant cells that accumulate proteinaceous reserves in the form of cyanophycin granules (Wetzel, 2001). These akinetes can survive low temperatures, desiccation and other adverse environmental conditions. When favourable conditions return, they germinate to produce trichomes. The ability of Nostocales to form akinetes confers a distinct advantage in environmental adaptation and subsequent bloom formation (Kim et al., 2005; Wetzel, 2001). C. raciborskii akinetes are more commonly observed in subtropical and temperate populations late in the population cycle as a component of seasonal population dynamics (Kravchuk et al., 2006). Particular set of physico-chemical conditions stimulate akinetes to germinate (Moore et al., 2005). Light has been implicated as well as temperature and nutrients as triggering factors (Huber, 1985). According to Moore (2004) germination of akinetes occurs at temperatures between 15 °C and 30 °C in tropical Australian strains of C. raciborskii. Mischke (2003) found filaments of this species at temperatures of about 17 oC in temperate lakes with those in the Scharmützelsee region in Germany. We therefore, tried to find out at what temperature the akinetes germinate in temperate region hypothesizing it should be below 15 oC. Since Aphanizomenon species like A. gracile and A. flosaquae are the most abundant natural Nostocales in that habitat, we compare it to the invaded C. raciborskii. To figure out the germination temperature for both species, field observations are complemented by experiments at different temperatures.