Planktothrix Blooms Research Articles

The recent disappearance of a persistent Planktothrix bloom: Characterization of a regime shift in the phytoplankton of Sandusky Bay (USA)

Sandusky Bay is the drowned mouth of the Sandusky River in the southwestern portion of Lake Erie. The bay is a popular recreation location and a regional source for drinking water. Like the western basin of Lake Erie, Sandusky Bay is known for being host to summer cyanobacterial harmful algal blooms (cHABs) year after year, fueled by runoff from the predominantly agricultural watershed and internal loading of legacy nutrients (primarily phosphorus). Since at least 2003, Sandusky Bay has harbored a microcystin-producing bloom of Planktothrix agardhii, a species of filamentous cyanobacteria that thrives in low light conditions. Long-term sampling (2003–2018) of Sandusky Bay revealed regular Planktothrix-dominated blooms during the summer months, but in recent years (2019–2022), 16S rRNA gene community profiling revealed that Planktothrix has largely disappeared. From 2017–2022, microcystin decreased well below the World Health Organization (WHO) guidelines. Spring TN:TP ratios increased in years following dam removal, yet there were no statistically significant shifts in other physicochemical variables, such as water temperature and water clarity. With the exception of the high bloom of Planktothrix in 2018, there was no statistical difference in chlorophyll during all other years. Concurrent with the disappearance of Planktothrix, Cyanobium spp. have become the dominant cyanobacterial group. The appearance of other potential toxigenic genera (i.e., Aphanizomenon, Dolichospermum, Cylindrospermopsis) may motivate monitoring of new toxins of concern in Sandusky Bay. Here, we document the regime shift in the cyanobacterial community and propose evidence supporting the hypothesis that the decline in the Planktothrix bloom was linked to the removal of an upstream dam on the Sandusky River.

Fate of Planktothrix-derived toxins in aquatic food webs: A case study in Lake Mindelsee (Germany)

Blooms of the red, filamentous cyanobacterium Planktothrix rubescens occur frequently in pre-alpine lakes in Europe, often with concomitant toxic microcystin (MC) production. Trophic transfer of MCs has been observed in bivalves, fish, and zooplankton species, while uptake of MCs into Diptera species could facilitate distribution of MCs into terrestrial food webs and habitats. In this study, we characterized a Planktothrix bloom in summer 2019 in Lake Mindelsee and tracked possible trophic transfer and/or bioaccumulation of MCs via analysis of phytoplankton, zooplankton (Daphnia) and emergent aquatic insects (Chaoborus, Chironomidae and Trichoptera). Using 16 S rRNA gene amplicon sequencing, we found that five sequence variants of Planktothrix spp. were responsible for bloom formation in September and October of 2019, and these MC-producing variants, provisionally identified as P. isothrix and/or P. serta, occurred exclusively in Lake Mindelsee (Germany), while other variants were also detected in nearby Lake Constance. The remaining cyanobacterial community was dominated by Cyanobiaceae species with high species overlap with Lake Constance, suggesting a well-established exchange of cyanobacteria species between the adjacent lakes. With targeted LC–HRMS/MS we identified two MC-congeners, MC-LR and [Asp3]MC-RR with maximum concentrations of 45 ng [Asp3]MC-RR/L in lake water in September. Both MC congeners displayed different predominance patterns, suggesting that two different MC-producing species occurred in a time-dependent manner, whereby [Asp3]MC-RR was clearly associated with the Planktothrix spp. bloom. We demonstrate an exclusive transfer of MC-LR, but not [Asp3]MC-RR, from phytoplankton into zooplankton reaching a 10-fold bioconcentration, yet complete absence of these MC congeners or their conjugates in aquatic insects. The latter demonstrated a limited trophic transfer of MCs from zooplankton to zooplanktivorous insect larvae (e.g., Chaoborus), or direct transfer into other aquatic insects (e.g. Chironomidae and Trichoptera), whether due to avoidance or limited uptake and/or rapid excretion of MCs by higher trophic emergent aquatic insects.

Microcystin profiles in European noble crayfish Astacus astacus and water in Lake Steinsfjorden, Norway

Lake Steinsfjorden, an important noble crayfish (Astacus astacus) habitat, is often affected by blooms of Planktothrix spp. that produce microcystins (MCs). A poor correlation between MCs by ELISA in the water and in crayfish tissue in a study in 2015 prompted further investigation by LC–HRMS. LC–HRMS analyses of filters from water samples and on selected crayfish tissue extracts from the 2015 study revealed the presence of known and previously unreported MCs. Crayfish samples from May and June 2015 were dominated by MCs from the Planktothrix bloom, whereas in September novel MCs that appeared to be metabolites of MC-LR were dominant, even though neither these nor MC-LR were detected in the water in 2015. A water sample from October 2016 also showed MCs typical of Planktothrix (i.e., [d-Asp3]- and [d-Asp3,Dhb7]MC-RR and -LR), but low levels of MC-RR and MC-LR were detected in the lake water for the first time. In late summer and autumn, the MC profiles of crayfish were dominated by the homonorvaline (Hnv) variant MC-LHnv, a putative metabolite of MC-LR. Taken together, ELISA, LC–HRMS and previous PCR analyses showed that although Planktothrix was part of the crayfish diet, it was not the sole source of MCs in the crayfish. Possibly, crayfish in Lake Steinsfjorden may be ingesting MCs from benthic cyanobacteria or from contaminated prey. Therefore, information on the cyanobacterial or MC content in the water column cannot safely be used to make predictions about MC concentrations in the crayfish in Lake Steinsfjorden. Interestingly, the results also show that targeted LC–MS analysis of the crayfish would at times have underestimated their MC content by nearly an order of magnitude, even if all previously reported MC variants had been included in the analysis.

Early and late cyanobacterial bloomers in a shallow, eutrophic lake.

Cyanobacterial blooms present challenges for water treatment, especially in regions like the Canadian prairies where poor water quality intensifies water treatment issues. Buoyant cyanobacteria that resist sedimentation present a challenge as water treatment operators attempt to balance pre-treatment and toxic disinfection by-products. Here, we used microscopy to identify and describe the succession of cyanobacterial species in Buffalo Pound Lake, a key drinking water supply. We used indicator species analysis to identify temporal grouping structures throughout two sampling seasons from May to October 2018 and 2019. Our findings highlight two key cyanobacterial bloom phases - a mid-summer diazotrophic bloom of Dolichospermum spp. and an autumn Planktothrix agardhii bloom. Dolichospermum crassa and Woronichinia compacta served as indicators of the mid-summer and autumn bloom phases, respectively. Different cyanobacterial metabolites were associated with the distinct bloom phases in both years: toxic microcystins were associated with the mid-summer Dolichospermum bloom and some newly monitored cyanopeptides (anabaenopeptin A and B) with the autumn Planktothrix bloom. Despite forming a significant proportion of the autumn phytoplankton biomass (>60%), the Planktothrix bloom had previously not been detected by sensor or laboratory-derived chlorophyll-a. Our results demonstrate the power of targeted taxonomic identification of key species as a tool for managers of bloom-prone systems. Moreover, we describe an autumn Planktothrix agardhii bloom that has the potential to disrupt water treatment due to its evasion of detection. Our findings highlight the importance of identifying this autumn bloom given the expectation that warmer temperatures and a longer ice-free season will become the norm.

Cyanobacterial blooms contribute to the diversity of antibiotic-resistance genes in aquatic ecosystems

Cyanobacterial blooms are a global ecological problem that directly threatens human health and crop safety. Cyanobacteria have toxic effects on aquatic microorganisms, which could drive the selection for resistance genes. The effect of cyanobacterial blooms on the dispersal and abundance of antibiotic-resistance genes (ARGs) of concern to human health remains poorly known. We herein investigated the effect of cyanobacterial blooms on ARG composition in Lake Taihu, China. The numbers and relative abundances of total ARGs increased obviously during a Planktothrix bloom. More pathogenic microorganisms were present during this bloom than during a Planktothrix bloom or during the non-bloom period. Microcosmic experiments using additional aquatic ecosystems (an urban river and Lake West) found that a coculture of Microcystis aeruginosa and Planktothrix agardhii increased the richness of the bacterial community, because its phycosphere provided a richer microniche for bacterial colonization and growth. Antibiotic-resistance bacteria were naturally in a rich position, successfully increasing the momentum for the emergence and spread of ARGs. These results demonstrate that cyanobacterial blooms are a crucial driver of ARG diffusion and enrichment in freshwater, thus providing a reference for the ecology and evolution of ARGs and ARBs and for better assessing and managing water quality.

Toward Disentangling the Multiple Nutritional Constraints Imposed by Planktothrix: The Significance of Harmful Secondary Metabolites and Sterol Limitation.

The harmful bloom-forming cyanobacterium Planktothrix is commonly considered to be nutritionally inadequate for zooplankton grazers, resulting in limited top-down control. However, interactions between Planktothrix and zooplankton grazers are poorly understood. The food quality of Planktothrix is potentially constrained by morphological properties (i.e., filament formation), the production of harmful secondary metabolites, and a deficiency in essential lipids (i.e., primarily sterols). Here, we investigated the relative significance of toxin production (microcystins, carboxypeptidase A inhibitors, protease inhibitors) and sterol limitation for the performance of Daphnia feeding on one Planktothrix rubescens and one P. agardhii wild-type/microcystin knock-out mutant pair. Our data suggest that the poor food quality of both Planktothrix spp. is due to deleterious effects mediated by various harmful secondary metabolites and that the impact of sterol limitation is partially or completely superimposed by toxicity. The significance of the different factors seems to depend on the metabolite profile of the considered Planktothrix strain and the Daphnia clone that is used for the experiments. The toxin-responsive gene expression (transporter genes, gpx, and trypsin) and enzyme activity patterns revealed strain-specific food quality constraints and that Daphnia is capable of modulating its physiological responses according to the ingested Planktothrix strain. Future studies need to consider that Planktothrix–grazer interactions are simultaneously modulated by multiple factors to improve our understanding of top-down influences on Planktothrix bloom formation.

Identification of Planktothrix (Cyanobacteria) Blooms and Effects on the Aquatic Macroinvertebrate Community in the Non-Tidal Potomac River, USA

Identification of Planktothrix (Cyanobacteria) Blooms and Effects on the Aquatic Macroinvertebrate Community in the Non-Tidal Potomac River, USA

Light stress in green and red Planktothrix strains: The orange carotenoid protein and its related photoprotective mechanism

Photosynthetic organisms need to sense and respond to fluctuating environmental conditions, to perform efficient photosynthesis and avoid the formation of harmful reactive oxygen species. Cyanobacteria have developed a photoprotective mechanism that decreases the energy arriving at the reaction centers by increasing thermal energy dissipation at the level of the phycobilisome, the extramembranal light-harvesting antenna. This mechanism is triggered by the photoactive orange carotenoid protein (OCP). In this study, we characterized OCP and the related photoprotective mechanism in non-stressed and light-stressed cells of three different strains of Planktothrix that can form impressive blooms. In addition to changing lake ecosystemic functions and biodiversity, Planktothrix blooms can have adverse effects on human and animal health as they produce toxins (e.g., microcystins). Three Planktothrix strains were selected: two green strains, PCC 10110 (microcystin producer) and PCC 7805 (non-microcystin producer), and one red strain, PCC 7821. The green strains colonize shallow lakes with higher light intensities while red strains proliferate in deep lakes. Our study allowed us to conclude that there is a correlation between the ecological niche in which these strains proliferate and the rates of induction and recovery of OCP-related photoprotection. However, differences in the resistance to prolonged high-light stress were correlated to a better replacement of damaged D1 protein and not to differences in OCP photoprotection. Finally, microcystins do not seem to be involved in photoprotection as was previously suggested.

Water quality and transparency characteristics during the Planktothrix abundance period in shallow Lake Kasumigaura, Japan

AbstractCyanobacterial blooms in lakes cause serious environmental problems on a global scale. Planktothrix (a filamentous cyanobacterium) blooms occurred during winter and spring from 2007 to 2011 in Lake Kasumigaura, a eutrophic lake located in Japan. This study analysed water quality during the period of Planktothrix abundance (2007–2010) at the centre of Lake Kasumigaura and compared it to that in the succeeding period, which did not have Planktothrix blooms (2012–2015). The average phytoplankton biovolume during the abundance period was larger than that in the succeeding period, which would contribute to the high chemical oxygen demand (CODMn; annual average of 9.7 mg/L) in the abundance period, compared to the succeeding period (7.6 mg/L). The total phosphorus concentration peaked in spring and summer in the abundance period, but only in summer in the succeeding period, whereas the seasonal variation in total nitrogen concentration between the two periods was relatively small. The annual average transparency (Secchi disc depth) increased before the abundance period due to a decrease in fixed suspended solid, which is comprised mainly of inorganic matter. It is likely that the change in irradiance conditions affected the occurrence of Planktothrix blooms.

Nitrogen cycling in Sandusky Bay, Lake Erie: oscillations between strong and weak export and implications for harmful algal blooms

Abstract. Recent global water quality crises point to an urgent need for greater understanding of cyanobacterial harmful algal blooms (cHABs) and their drivers. Nearshore areas of Lake Erie such as Sandusky Bay may become seasonally limited by nitrogen (N) and are characterized by distinct cHAB compositions (i.e., Planktothrix over Microcystis). This study investigated phytoplankton N uptake pathways, determined drivers of N depletion, and characterized the N budget in Sandusky Bay. Nitrate (NO3-) and ammonium (NH4+) uptake, N fixation, and N removal processes were quantified by stable isotopic approaches. Dissimilatory N reduction was a relatively modest N sink, with denitrification, anammox, and N2O production accounting for 84, 14, and 2 % of sediment N removal, respectively. Phytoplankton assimilation was the dominant N uptake mechanism, and NO3- uptake rates were higher than NH4+ uptake rates. Riverine N loading was sometimes insufficient to meet assimilatory and dissimilatory demands, but N fixation alleviated this deficit. N fixation made up 23.7–85.4 % of total phytoplankton N acquisition and indirectly supports Planktothrix blooms. However, N fixation rates were surprisingly uncorrelated with NO3- or NH4+ concentrations. Owing to temporal separation in sources and sinks of N to Lake Erie, Sandusky Bay oscillates between a conduit and a filter of downstream N loading to Lake Erie, delivering extensively recycled forms of N during periods of low export. Drowned river mouths such as Sandusky Bay are mediators of downstream N loading, but climate-change-induced increases in precipitation and N loading will likely intensify N export from these systems.

Low temperatures can promote cyanobacterial bloom formation by providing refuge from microbial antagonists.

Freshwater cyanobacteria are prone to a wide range of highly potent microbial antagonists. Most of these exploit their prey in a frequency-dependent manner and are therefore particularly well suited to prevent any accumulation of cyanobacteria. Mass developments of cyanobacteria, the so-called blooms, should therefore be rare events, which is in striking contrast to what we actually see in nature. Laboratory experiments of the present study showed that the temperature range 5.8–10 °C forms a thermal refuge, inside which the cyanobacterium Planktothrix can grow without being exploited by two otherwise highly potent microbial antagonists. In nature, access of Planktothrix to this refuge was associated with positive net growth and a high probability of bloom formation, confirming that refuge temperatures indeed allow Planktothrix to grow with a minimum of biomass loss caused by microbial antagonists. Contact to higher temperatures, in contrast, was associated with decreases in net growth rate and in probability of bloom formation, with population collapses and with the occurrence of parasite infection. This is in agreement with the finding of laboratory experiments that above 10 °C exploitation of Planktothrix by multiple microbes increases in a temperature-dependent manner. Taken together, above findings suggest that temperature modulates the microbial control of natural Planktothrix populations. Low temperatures form a thermal refuge that may promote Planktothrix bloom formation by shielding the cyanobacterium from otherwise highly potent microbial antagonists.

Environmental Conditions Determine the Course and Outcome of Phytoplankton Chytridiomycosis.

Chytrid fungi are highly potent parasites of phytoplankton. They are thought to force phytoplankton organisms into an evolutionary arms race with high population diversity as the outcome. The underlying selection regime is known as Red Queen dynamics. However, our study suggests a more complex picture for chytrid parasitism in the cyanobacterium Planktothrix. Laboratory experiments identified a “cold thermal refuge”, inside which Planktothrix can grow without chytrid infection. A field study in two Norwegian lakes underlined the ecological significance of this finding. The study utilized sediment DNA as a biological archive in combination with existing monitoring data. In one lake, temperature and light conditions forced Planktothrix outside the thermal refuge for most of the growing season. This probably resulted in Red Queen dynamics as suggested by a high parasitic pressure exerted by chytrids, an increase in Planktothrix genotype diversity over time, and a correlation between Planktothrix genotype diversity and duration of bloom events. In the second lake, a colder climate allowed Planktothrix to largely stay inside the thermal refuge. The parasitic pressure exerted by chytrids and Planktothrix genotype diversity remained low, indicating that Planktothrix successfully evaded the Red Queen dynamics. Episodic Planktothrix blooms were observed during spring and autumn circulation, in the metalimnion or under the ice. Interestingly, both lakes were dominated by the same or related Planktothrix genotypes. Taken together, our data suggest that, depending on environmental conditions, chytrid parasitism can impose distinct selection regimes on conspecific phytoplankton populations with similar genotype composition, causing these populations to behave and perhaps to evolve differently.

Effects of nutrients and warming on Planktothrix dynamics and diversity: a palaeolimnological view based on sedimentary DNA and RNA

Summary Decennial changes in Planktothrix rubescens diversity and dynamics were reconstructed by applying molecular tools to analyse DNA and RNA extracted from lake sediments. The sediments studied were sampled from a deep peri‐alpine lake that has experienced both dramatic shifts in trophic conditions and large‐scale climatic changes. Palaeolimnological proxies were combined with statistical modelling to investigate the relative influence of phosphorus concentrations and temperature changes on the extent of Planktothrix blooms over the last century. Phylogenetic analysis revealed that the overall composition of the cyanobacterial community changed over the transition from oligotrophic to eutrophic conditions. When the relative abundance of Planktothrix decreased in the 1970s, concomitant with eutrophication, total cyanobacterial abundance remained high and more Anabaena and Microcystis sequences were detected. In spite of such drastic environmental changes, the lake provided a constant niche for one particular Planktothrix species, which was consistently present from the 1920s to the present day. Phosphorus concentration was found to be the dominant driver of the relative abundance of P. rubescens, with the highest abundances observed during mesotrophic conditions. The relative role of climate was nutrient‐dependent, with warmer springs having a positive effect on P. rubescens abundance only during mesotrophic periods. Overall, this study confirms that analysis of genetic signatures preserved in sediment archives allows assessment of key palaeoenvironmental indicator species that have no diagnostic microscopic cellular features in the sediment record. In the case of cyanobacteria, palaeogenetics offer unique opportunities to anticipate how future climate change might affect the response of P. rubescens to phosphorus concentration.

Organic and inorganic nitrogen utilization by nitrogen-stressed cyanobacteria during bloom conditions

Cyanobacterial blooms often occur in lakes that have high phosphorus (P) and low nitrogen (N) concentrations, and the growth rate of the blooms is often constrained by N. For these reasons, many researchers have suggested that regulation of both P and N is required to control eutrophication. However, because N occurs in many bioavailable forms, regulation of a particular form may be beneficial rather than regulation of all N forms. To address how N-stressed cyanobacteria respond to various N inputs, N enrichment experiments (nitrate, ammonium, urea, and alanine) were performed during N-limited cyanobacterial blooms in Maumee and Sandusky Bays of Lake Erie and in Grand Lake St. Marys (GLSM). Bioavailable N (nitrate, urea, and ammonium) concentrations were also determined. Microcystis aeruginosa dominated the Maumee Bay bloom, where the highest growth rates were in response to ammonium additions, and lowest growth rates were in response to nitrate. Urea and the amino acid alanine resulted in intermediate growth rates. Planktothrix agardhii dominated the Sandusky Bay and GLSM blooms, where nitrate, ammonium, and urea addition resulted in similar growth rates. Additions of alanine did not stimulate growth of the Planktothrix blooms. Incubations using stable isotope 15N showed the cyanobacteria had a preference for ammonium, but the other forms were also assimilated in the presence of ammonium. These results show that cyanobacterial blooms will assimilate multiple forms of N to support growth. Thus, if lake managers do decide that N abatement is necessary, then all forms of bioavailable N need to be constrained.

Effects of a toxic cyanobacterial bloom (Planktothrix agardhii) on fish: Insights from histopathological and quantitative proteomic assessments following the oral exposure of medaka fish (Oryzias latipes)

Cyanobacterial toxic blooms often occur in freshwater lakes and constitute a potential health risk to human populations, as well as to fish and other aquatic organisms. Microcystin-LR (the cyanotoxin most commonly detected in the freshwater environment) is a potent hepatotoxin, deregulating the kinase pathway by inhibiting phosphatases 1 and 2A. Although toxicological effects have been clearly linked to the in vitro exposure of fish to purified microcystins, cyanotoxins are produced by the cyanobacteria together with numerous other potentially toxic molecules, and their overall and specific implications for the health of fish have still not been clearly established and remain puzzlingly difficult to assess.The medaka fish (Oryzias latipes) was chosen as an in vitro model for studying the effects of a cyanobacterial bloom on liver protein contents using a gel free quantitative approach, iTRAQ, in addition to pathology examinations on histological preparations. Fish were gavaged with 5μL cyanobacterial extracts (Planktothrix agardhii) from a natural bloom (La Grande Paroisse, France) containing 2.5μgequiv. MC-LR. 2h after exposure, the fish were sacrificed and livers were collected for analysis. Histological observations indicate that hepatocytes present glycogen storage loss, and cellular damages, together with immunological localization of MCs. Using a proteomic approach, 304 proteins were identified in the fish livers, 147 of them with a high degree of identification confidence. Fifteen of these proteins were statistically significantly different from those of controls (gavaged with water only). Overall, these protein regulation discrepancies clearly indicate that oxidative stress and lipid regulation had occurred in the livers of the exposed medaka fish. In contrast to previous pure microcystin-LR gavage experiments, marked induction of vitellogenin 1 protein was observed for the first time with a cyanobacterial extract. This finding was confirmed by ELISA quantification of vitellogenin liver content, suggesting that the Planktothrix bloom extract had induced the occurrence of an endocrine-disrupting effect.

Rarity, ecological memory, rate of floral change in phytoplankton—and the mystery of the Red Cock

In this article, we attempt to estimate the contemporary phytoplankton species pool of a particular lake, by assessing the rate of floral change over a period of 15 years. Phytoplankton time series data from Lake Stechlin, an oligo-mesotrophic lake in the Baltic Lake District (Germany) were used. Of the 254 algal species recorded during the 15-year of studies with roughly biweekly sampling, 212 species were planktonic. In the individual plankton years, the recorded total number of species changed between 97 and 122, of which the number of dominants (>1% contribution to the annual average of total biomass) was only 10–19. The 15-year cumulative number of species exhibited an almost linear increase after an initial saturation phase. This increase was attributed to two reasons: increase of sample size and immigration of species new to the flora. Based on a probabilistic model developed in this study, we estimated the number of co-existing planktonic species of the lake as some 180, and the rate of floral change as 1–2 species per year. Of these co-existing species, only few maintain the matter–energy processing ecosystem functions in any particular plankton year. Selection of these dominants is probably driven by mesoclimatic cycles, coupled with human-induced forcing, like eutrophication. All others are hiding as an ecological memory, in the sense of the capacity or experiences of past states to influence present or future responses of the community. Data analyses suggest that selection of the ‘memory species’ that show temporary abundance increases over shorter (several years) periods are largely dependent upon the dominants. These results show that interspecific interactions and the particular autecological features of the dominants, together with their effects on the whole ecosystem, act as a major organizing force. Some phytoplankton species, like Planktothrix rubescens, are efficient ecosystem engineers with cascading effects of both a top-down and bottom-up nature. Historical scientific data on Planktothrix blooms in Lake Stechlin suggest cyclic patterns in long-term development of phytoplankton which, as the legend of the Red Cock suggests, dates back much further than scientific archives.

Physiological stress and pathology in European whitefish ( Coregonus lavaretus) induced by subchronic exposure to environmentally relevant densities of Planktothrix rubescens

Planktothrix rubescens belongs to the most ubiquitous cyanobacterial species in mesotrophic and oligotrophic lakes in the pre-alpine regions. In most of these lakes, coregonids are among the dominant species of the ichthyofauna with great importance for the professional fishery. A possible link between the occurrence of toxic Planktothrix blooms and the recurrent slumps in coregonid yields has been suggested. Indeed, acute toxic effects of microcystins and other cyanobacterial toxins have been shown for various fish species. However, chronic exposure scenarios appear to be more common and thus more environmentally realistic than acute intoxications. The aim of this study was therefore to investigate the physiological stress response and organ pathology in coregonids sub-chronically exposed to ambient water containing low, medium and high P. rubescens densities, known to be typical of pre-alpine lakes. Coregonid hatchlings were exposed in four tanks containing 0 (sham-control) and approximately 1500 (low), 15,000 (medium) and 55,000 (high) P. rubescens cells/ml for up to 28 days. Temperature, oxygen concentration, pH-value, P. rubescens cell density and microcystin concentration were recorded and the fish were observed for behavioural changes and examined for parasite infestations. Gill ventilation rates, general condition factors and mortalities were determined and liver, kidney, gut and gill were assessed histopathologically and immunhistologically. Depending on the cell density, exposed fish showed behavioural changes, including increased ventilation rates possibly representing a physiological stress response. Susceptibility to ectoparasitic infestation and increased mortality in exposed fish suggested P. rubescens associated effects on fish fitness. Histopathological alterations in liver, gastrointestinal tract and kidney, which were also immunopositive for microcystin suggested causality of tissue damage and the presence of microcystins. In contrast, observed gill pathology appeared to result primarily from mechanical abrasion and irritation due to ectoparasitic infestation. The current exposure experiment confirmed the hypothesis that subchronic and chronic exposure to low cyanobacterial cell densities and hence microcystins can exacerbate physiological stress and sustained pathological alterations in exposed coregonids. The study therefore supports the theory that P. rubescens blooms may be causal to the observed weight reduction and hence fitness of coregonids in pre-alpine lakes such as Lake Ammersee (Germany).

Toxicity and microcystin content of extracts from a Planktothrix bloom and two laboratory strains

Toxic effects and microcystin content from various extracts of a Planktothrix agardhii bloom and two different strains of Planktothrix agardhii, HUB 076, and NIVA 34 were investigated. Extracts were obtained with solvents of different polarity such as hexane, dichloromethane, methanol, and water. Additionally, different pre-treatments were used to break the cells before extraction. Acute toxicity was determined with the fairy shrimp Thamnocephalus platyurus, subchronic effects were detected in embryos and larvae of the zebrafish Danio rerio. The extracts affected the test species to a different extent. Effects were severe in polar extracts (water and methanol) of all strains tested. Although the strain NIVA 34 did not contain any microcystins, aqueous extracts of this strain showed the highest acute toxicity to the crustacean species tested (LC 50=0.08 mg ml −1). In contrast aqueous extracts of the Planktothrix bloom containing high amounts of microcystins were less acutely toxic (LC 50=0.46 mg ml −1). Our results indicate the existence of further toxic metabolites in different Planktothrix agardhii strains.

Presence of Planktothrix sp. and cyanobacterial toxins in Lake Ammersee, Germany and their impact on whitefish (Coregonus lavaretus L.).

Due to the increasing oligotrophy of Lake Ammersee, southern Germany, metalimnic Planktothrix have become one of the dominant planktonic species causing regular blooms. Whitefish (Coregonus lavaretus) is the dominant local fish species with great importance for the fishing industry. Recently, whole age groups of this fish species have disappeared and since 1991, average body-weight has decreased. The causes for this remain unclear. Planktothrix species produce the cyclic peptide toxin desmethyl microcystin-RR, which inhibits glycogen metabolism and has detrimental effects on the development of aquatic organisms. During blooms, gut contents of whitefish displayed a blue discoloration, possibly representing phycobiliproteins typical for cyanobacteria. This study aimed to elucidate the impact of Planktothrix blooms on fish population dynamics. Planktothrix cell counts, performed by epifluorescence microscopy, showed blooms to contain up to 80,000 cells/ml. Microcystin levels of 1-5 micrograms/mg dry weight in Planktothrix extracts and 0.08 microgram/l in water samples, were determined via HPLC and protein-phosphatase inhibition assay. Planktothrix filaments were detected in gut contents of whitefish. In addition, microcystins could be detected in gut contents via ELISA. Similarly, immunoprobing with microcystin-antibodies demonstrated microcystin-protein adducts in liver homogenates of whitefish caught during Planktothrix blooms. Furthermore, Planktothrix extracts proved cytotoxic to trout hepatocytes as determined by MTT reduction. Whitefish eggs and larvae, obtained during blooms in winter 1998 and 2000 in a Lake Ammersee hatchery, were studied for developmental progress. Malformations typical for cyanobacterial toxin exposure were observed.