Microbial community structures and bacteria-Cylindrospermopsis raciborskii interactions in Yilong Lake.

Cylindrospermopsis raciborskii-dominated harmful algae blooms have been reported globally in recent years. However, our understanding of the ecology of C. raciborskii in natural conditions is still poor. In this study, we collected the water samples from a C. raciborskii-blooming lake, Yilong Lake, in Yunnan province, China, and used both culture-dependent and culture-independent approaches to investigate their microbial communities and the interactions between C. raciborskii and the other bacteria. The composition and diversity of microbial communities were revealed with 16S rRNA gene high-throughput sequencing data analysis. Microbial co-occurrences analysis suggests C. raciborskii may have complex associations with other bacteria. Based on co-inoculation tests, we obtained 14 strains of bacterial strains from the water samples that exhibited either algicidal or promoting effects on a strain of C. raciborskii. Two bacterial isolates exhibited a consistent performance between co-occurrence analysis and experimental results. Effects of these bacteria-algae interspecies interactions on the bloom event are discussed. All these results may provide new insights into the C. raciborskii-dominated blooms and how its interspecies relationships with other bacteria may influence the bloom events in eutrophic waters throughout the world.

Ecophysiological Aspects and sxt Genes Expression Underlying Induced Chemical Defense in STX-Producing Raphidiopsis raciborskii (Cyanobacteria) against the Zooplankter Daphnia gessneri.

Cyanobacteria stand out among phytoplankton when they form massive blooms and produce toxins. Because cyanotoxin genes date to the origin of metazoans, the hypothesis that cyanotoxins function as a defense against herbivory is still debated. Although their primary cellular function might vary, these metabolites could have evolved as an anti-predator response. Here we evaluated the physiological and molecular responses of a saxitoxin-producing Raphidiopsis raciborskii to infochemicals released by the grazer Daphnia gessneri. Induced chemical defenses were evidenced in R. raciborskii as a significant increase in the transcription level of sxt genes, followed by an increase in saxitoxin content when exposed to predator cues. Moreover, cyanobacterial growth decreased, and no significant effects on photosynthesis or morphology were observed. Overall, the induced defense response was accompanied by a trade-off between toxin production and growth. These results shed light on the mechanisms underlying zooplankton-cyanobacteria interactions in aquatic food webs. The widespread occurrence of the cyanobacterium R. raciborskii in freshwater bodies has been attributed to its phenotypic plasticity. Assessing the potential of this species to thrive over interaction filters such as zooplankton grazing pressure can enhance our understanding of its adaptive success.

Comparative proteomics of the toxigenic diazotroph Raphidiopsis raciborskii (cyanobacteria) in response to iron.

Raphidiopsis raciborskii is an invasive bloom-forming cyanobacteria with the flexibility to utilize atmospheric and fixed nitrogen. Since nitrogen-fixation has a high requirement for iron as an ezyme cofactor, we hypothesize that iron availability would determine the success of the species under nitrogen-fixing conditions. This study compares the proteomic response of cylindrospermopsin-producing and non-toxic strains of R. racibroskii to reduced iron concentrations, under nitrogen-fixing conditions, to examine any strain-specific adaptations that might increase fitness under these conditions. We also compared their proteomic responses at exponential and stationary growth phases to capture the changes throughout the growth cycle. Overall, the toxic strain was more competitive under Fe-starved conditions during exponential phase, with upregulated growth and transport-related proteins. The non-toxic strain showed reduced protein expression across multiple primary metabolism pathways. We propose that the increased expression of porin proteins during the exponential growth phase enables toxic strains to persist under Fe-starved conditions with this ability providing a potential explanation for the increased fitness of cylindrospermoipsin-producing strains during unfavourable environmental conditions.

Effects and possible mechanisms of sanguinarine on the competition between Raphidiopsis raciborskii (Cyanophyta) and Scenedesmus obliquus (Chlorophyta): A comparative toxicological study.

The phytogenic algicide sanguinarine shows strong inhibitory effects on some bloom-forming cyanobacteria and exhibits great potential in cyanobacterial bloom mitigation. To evaluate the possible ecological effects of sanguinarine on microalgae, the effects and possible mechanisms of sanguinarine on the competition between bloom-forming cyanobacterium Raphidiopsis raciborskii (formerly named Cylindrospermopsis raciborskii) and green alga Scenedesmus obliquus were investigated through co-culture competition test and comparative toxicological study including growth characteristics, chlorophyll fluorescence transients, activities of antioxidant enzymes, and lipid peroxidation. The results of Raphidiopsis-Scenedesmus co-culture competition test showed that sanguinarine decreased the competition ability of R. raciborskii, which benefitted S. obliquus in winning the competition. Toxicological studies have shown that sanguinarine exhibited high inhibitory effects on the growth and photosynthesis of R. raciborskii but no obvious toxicity on S. obliquus at concentrations of no more than 80 µg L-1. Oxidative damage partially contributed but was not the primary mechanism for the toxicity of sanguinarine on R. raciborskii. The results presented in this study indicate that sanguinarine may be a good algicidal candidate in mitigation of Raphidiopsis-based water bloom.

In Vitro Toxicological Screening of Stable and Senescing Cultures of Aphanizomenon, Planktothrix, and Raphidiopsis.

Toxicity of cyanobacteria is the subject of ongoing research, and a number of toxic metabolites have been described, their biosynthesis pathways have been elucidated, and the mechanism of their action has been established. However, several knowledge gaps still exist, e.g., some strains produce hitherto unknown toxic compounds, while the exact dynamics of exerted toxicity during cyanobacterial growth still requires further exploration. Therefore, the present study investigated the toxicity of extracts of nine freshwater strains of Aphanizomenon gracile, an Aphanizomenon sp. strain isolated from the Baltic Sea, a freshwater strain of Planktothrix agardhii, and two strains of Raphidiopsis raciborskii obtained from 25- and 70-day-old cultures. An in vitro experimental model based on Cyprinus carpio hepatocytes (oxidative stress markers, DNA fragmentation, and serine/threonine protein activity) and brain homogenate (cholinesterase activity) was employed. The studied extracts demonstrated toxicity to fish cells, and in general, all examined extracts altered at least one or more of considered parameters, indicating that they possess, to some degree, toxic potency. Although the time from which the extracts were obtained had a significant importance for the response of fish cells, we observed strong variability between the different strains and species. In some strains, extracts that originated from 25-day-old cultures triggered more harmful effects on fish cells compared to those obtained from 70-day-old cultures, whereas in other strains, we observed the opposite effect or a lack of a significant change. Our study revealed that there was no clear or common pattern regarding the degree of cyanobacterial bloom toxicity at a given stage of development. This means that young cyanobacterial blooms that are just forming can pose an equally toxic threat to aquatic vertebrates and ecosystem functioning as those that are stable or old with a tendency to collapse. This might be largely due to a high variability of strains in the bloom.

Clethodim (herbicide) alters the growth and toxins content of Microcystis aeruginosa and Raphidiopsis raciborskii.

Increased agricultural intensification goes with the widespread use of herbicides that adversely affect aquatic biodiversity. The effects of herbicides on toxin-producing cyanobacteria have been poorly studied. The present study aimed to investigate the toxicological and physiological effects of the herbicide clethodim on Raphidiopsis raciborskii (a.k.a. Cylindrospermopsis raciborskii) ITEPA1 and Microcystis aeruginosa BCCUSP232. On day four of the experiment, the exposure to 25 mg/L clethodim resulted in the highest cell density of R. raciborskii. Similarly, exposure to the 1, 5, 20, and 50 mg/L clethodim treatments resulted in the highest cell densities of M. aeruginosa on day 4 of the experiment. Medium effect concentrations (EC50) after 96 h of exposure of both strains to clethodim were 192.98 mg/L and 168.73 mg/L for R. raciborskii and M. aeruginosa, respectively. The presence of clethodim significantly increased the total microcystin content of M. aeruginosa compared to the control cultures. At 400 mg/L, total saxitoxins content of R. raciborskii was 27% higher than that of the control cultures on day 4. In contrast, cultures exposed to 100 mg/L clethodim had the lowest saxitoxins levels per cell quota. There was an increase in the levels of intracellular hydrogen peroxide in both species during exposure to clethodim, which was followed by significant changes (p < 0.05) in the activity of antioxidant enzymes such as peroxidase and superoxide dismutase. These results revealed that the presence of low levels of clethodim in the aquatic environment might lead to the excessive proliferation of cyanobacteria and alteration of their cyanotoxins content.

Low invasion success of an invasive cyanobacterium in a chlorophyte dominated lake.

Biological invasions are a major threat to biodiversity and ecosystem functioning. Successful invasions depend on the interplay of multiple abiotic and biotic factors, however, the process of the invasion itself is often overlooked. The temporal variation of environmental factors suggests that a 'window of opportunity' for successful invasions exists. Especially aquatic habitats, like temperate lakes, undergo pronounced seasonal fluctuations and show temporally varying environmental conditions in e.g. nutrient availability, temperature and the composition of the resident community including competitors and consumers. We experimentally tested if an invasion window for the globally invasive cyanobacterium Cylindrospermopsis raciborskii exists. From May to September, we determined the invasion success of C. raciborskii in laboratory mesocosms with natural lake water. Although the invasion success was generally low, the invasiveness varied among months and differed in total invasive biomass, net development and final share of C. raciborskii in the community. During the first days, C. raciborskii strongly declined and this initial, short-term decline was independent of the ambient consumptive pressure. These results are in contrast to laboratory studies in which C. raciborskii successfully invaded, suggesting that a complex natural system develops a resistance to invasions.

Combined Effect of Light and Temperature on the Production of Saxitoxins in Cylindrospermopsis raciborskii Strains.

Cylindrospermopsis raciborskii is a potentially toxic freshwater cyanobacterium that can tolerate a wide range of light and temperature. Due to climatic changes, the interaction between light and temperature is studied in aquatic systems, but no study has addressed the effect of both variables on the saxitoxins production. This study evaluated the combined effect of light and temperature on saxitoxins production and cellular quota in C. raciborskii. Experiments were performed with three C. raciborskii strains in batch cultures under six light intensities (10, 40, 60, 100, 150, and 500 µmol of photons m-2 s-1) and four temperatures (15, 20, 25, and 30 °C). The growth of C. raciborskii strains was limited at lower temperatures and the maximum growth rates were obtained under higher light combined with temperatures equal or above 20 °C, depending on the strain. In general, growth was highest at 30 °C at the lower light intensities and equally high at 25 °C and 30 °C under higher light. Highest saxitoxins concentration and cell-quota occurred at 25 °C under high light intensities, but were much lower at 30 °C. Hence, increased temperatures combined with sufficient light will lead to higher C. raciborskii biomass, but blooms could become less toxic in tropical regions.

Variability in a permanent cyanobacterial bloom: species-specific responses to environmental drivers.

Cyanobacterial blooms are characterized by intense growth of one or few species that will dominate the phytoplankton community for periods of few months to an entire year or more. However, even during persistent blooms, important seasonal changes among dominant species can be observed. Pampulha reservoir is a tropical eutrophic reservoir presenting permanent blooms. To identify the main species in this environment, a closer analysis performed by microscopy and 16S-rRNA DGGE revealed Cylindrospermopsis raciborskii as highly dominant throughout the year. The second most abundant group comprised species belonging to the Microcystis genus. They followed a well-defined seasonal pattern described by interesting species-specific ecological trends. During thermal stratification in the rainy/warmer season, C. raciborskii dominated in the water column, while Microcystis spp. were abundant at the end of the dry season, a period characterized by higher total phosphorus concentrations. Phylogenetic analyses confirmed the two dominant taxa and their seasonal trends. The results showed that cyanobacteria major controlling factors were strongly species dependent, shifting from physical/climate related (stratification) to more chemical driven (nutrients/eutrophication). Identifying these drivers is therefore essential for the understanding of the bloom dynamics and the real risks associated with each species, and to eventually adopt the most appropriate and effective management strategies.

Hydrogen peroxide treatment promotes chlorophytes over toxic cyanobacteria in a hyper-eutrophic aquaculture pond.

Controlling blooms of toxigenic phytoplankton, including cyanobacteria, is a high priority for managers of aquatic systems that are used for drinking water, recreation, and aquaculture production. Although a variety of treatment approaches exist, hydrogen peroxide (H2O2) has the potential to be an effective and ecofriendly algaecide given that this compound may select against cyanobacteria while not producing harmful residues. To broadly evaluate the effectiveness of H2O2 on toxigenic phytoplankton, we tested multiple concentrations of H2O2 on (1) four cyanobacterial cultures, including filamentous Anabaena, Cylindrospermopsis, and Planktothrix, and unicellular Microcystis, in a 5-day laboratory experiment and (2) a dense cyanobacterial bloom in a 7-day field experiment conducted in a nutrient-rich aquaculture pond. In the laboratory experiment, half-maximal effective concentrations (EC50) were similar for Anabaena, Cylindrospermopsis, and Planktothrix (average EC50 = 0.41 mg L-1) but were ∼10x lower than observed for Microcystis (EC50 = 5.06 mg L-1). Results from a field experiment in an aquaculture pond showed that ≥1.3 and ≥ 6.7 mg L-1 of H2O2 effectively eliminated Planktothrix and Microcystis, respectively. Moreover, 6.7 mg L-1 of H2O2 reduced microcystin and enhanced phytoplankton diversity, while causing relatively small negative effects on zooplankton abundance. In contrast, 20 mg L-1 of H2O2 showed the greatest negative effect on zooplankton. Our results demonstrate that H2O2 can be an effective, rapid algaecide for controlling toxigenic cyanobacteria when properly dosed.

Chlorophyll a fluorescence and transcriptome reveal the toxicological effects of bisphenol A on an invasive cyanobacterium, Cylindrospermopsis raciborskii.

Bisphenol A has attracted worldwide attention due to its harmful effects on humans, animals and plants. In this study, the toxicological effects of BPA on Cylindrospermopsis raciborskii were assessed based on chlorophyll a fluorescence and transcriptome analyses. The results showed that the growth of C. raciborskii was significantly inhibited when BPA exceeded 0.1 mg L-1. A marked rise of phase J was observed at a concentration greater than 0.1 mg L-1, while a K phase appeared at 20 mg L-1. The chlorophyll a fluorescence parameters of RC/CS0, F0, φP0, φE0, and ψ0, underwent a significant decline under all treatments of BPA, whereas a significant increase in both VJ and M0 occurred under all concentrations of BPA. Additionally, ABS/RC and DIo/RC markedly increased at 10 mg L-1 and 20 mg L-1. The transcriptome analysis revealed that the genes of photosynthesis, including psbA, psbB, psbC, psbD, apcA, apcB, cpcA, and cpcB, as well as those of chlorophyll and carotenoid biosynthesis, namely hemN, acsF, chlL, chlN, chlP, crtB, pds, were all down-regulated. Moreover, BPA also inhibited the oxidative phosphorylation, glycolysis/gluconeogenesis, citrate cycle (TCA cycle), and fatty acid metabolism in C. raciborskii. Taken together, these results suggest BPA can negatively affect the expression of multiple genes and the vital energy metabolism process to arrest the growth and photosynthesis of C. raciborskii.

Effects of polyaluminum chloride and lanthanum-modified bentonite on the growth rates of three Cylindrospermopsis raciborskii strains.

In tropical and subtropical lakes, eutrophication often leads to nuisance blooms of Cylindrospermopsis raciborskii. In laboratory experiments, we tested the combined effects of flocculant polyaluminum chloride (PAC) and lanthanum-modified bentonite (LMB) on the sinking and growth rates of three C. raciborskii strains. We tested the hypothesis that the combination of PAC and LMB would (1) effectively sink C. raciborskii in a test tube experiment and (2) impair C. raciborskii growth, irrespective of the biomass of the inoculum (bloom) and the strain in the growth experiment. We tested the recommended (LMB1) and a three-times higher dose of LMB (LMB3). The combined addition of PAC and LMB enhanced the sedimentation of all C. raciborskii strains. Moreover, both the PAC and LMB doses decreased the phosphate concentration. PAC and LMB1 decreased the growth rate of all strains, but the efficacy depended on the biomass and strain. The combined addition of PAC and LMB3 inhibited the growth of all strains independently of the biomass and strain. We conclude that a low dose of PAC in combination with the recommended dose of LMB decreases C. raciborskii blooms and that the efficiency of the technique depends on the biomass of the bloom. A higher dose of LMB is needed to obtain a more efficient control of C. raciborskii blooms.

[Distribution and Factors Affecting Cylindrospermopsis raciborskii in Guangdong Reservoirs].

Cylindrospermopsis raciborskii originating from tropical and subtropical regions is potentially toxic and attracts much attention due to its extension to the global temperate zone in recent years. Based on historical data of 20 reservoirs with different trophic levels (dry season, wet season, and transitional season of 2010), this study focuses on the analysis of the occurrence and distribution of C. raciborskii in the Guangdong Province. Based on the results, C. raciborskii was found in 19 of the 20 reservoirs and its biomass ranges from 0.0001-39.740 mg·L-1 and accounts for 0.02%-97.07% of the total phytoplankton biomass. Both a notable spatial and seasonal distribution of C. raciborskii were observed. Its occurrence is higher in the western coastal area (77.78%) than in the Zhujiang Delta (66.67%) and northern coastal area (33.33%) and is relatively lower in the dry season (40%) compared with the rainy season (70%) and transition season (85%). The trophic level has a significant effect on the presence of C. raciborskii, which is notably higher in eutrophic reservoirs (81.48%) than in mesotrophic reservoirs (66.67%) and oligotrophic reservoirs (33.33%). The redundancy analysis shows that C. raciborskii biomass is positively correlated with total nitrogen (TN) and the trophic state index (TSI) and negatively correlated with dissolved inorganic nitrogen (DIN), soluble reactive phosphorus (SRP), and the secchi depth (SD). Thus, C. raciborskii in Guangdong reservoirs may be promoted by environmental factors such as high nitrogen contents, low phosphorus concentration, and transparency.

The Effect of Bisphenol A on Growth, Morphology, Lipid Peroxidation, Antioxidant Enzyme Activity, and PS II in Cylindrospermopsis raciborskii and Scenedesmus quadricauda.

To investigate the effect of bisphenol A (BPA) on Cylindrospermopsis raciborskii (Cyanobacteria) and Scenedesmus quadricauda (Chlorophyta), we grew the two species at BPA concentrations of 0, 0.1, 1, 2, 5, 10, and 20 mg/L and examined their growth, lipid peroxidation, antioxidant enzyme activity, and chlorophyll a fluorescence. The 96-h EC50 values (effective concentration causing 50% growth inhibition) for BPA in C. raciborskii and S. quadricauda were 9.663 ± 0.047, and 13.233 ± 0.069 mg/L, respectively. A significant reduction in chlorophyll a concentration was found in C. raciborskii and S. quadricauda when BPA concentrations were greater than 1 and 2 mg/L, respectively. Furthermore, F v/F m, ΔF/F m', and qP decreased significantly at 10 mg/L BPA in C. raciborskii but started to decrease at 10 mg/L in S. quadricauda. The changes in chlorophyll fluorescence parameters (α, rETRmax) that were obtained from the rapid light response curves of both algae species showed similar responses to F v/F m, ΔF/F m', and qP under BPA-induced stress. Values for all of the chlorophyll fluorescence parameters in S. quadricauda were higher than in C. raciborskii; however, the nonphotochemical quenching measured in C. raciborskii was considerably higher than it was in S. quadricauda. In addition, lipid peroxidation (determined as MDA content) and antioxidant enzyme activities (SOD and CAT) increased in both species as the BPA concentration increased. These results suggest that C. raciborskii is more sensitive to the effects of BPA than S. quadricauda and that photosystem II might be a target for the activity of BPA in vivo.

Variation within and between cyanobacterial species and strains affects competition: Implications for phytoplankton modelling.

Cyanobacteria Microcystis aeruginosa and Cylindrospermopsis raciborskii are two harmful species which co-occur and successively dominate in freshwaters globally. Within-species strain variability affects cyanobacterial population responses to environmental conditions, and it is unclear which species/strain would dominate under different environmental conditions. This study applied a Monte Carlo approach to a phytoplankton dynamic growth model to identify how growth variability of multiple strains of these two species affects their competition. Pairwise competition between four M. aeruginosa and eight C. raciborskii strains was simulated using a deterministic model, parameterized with laboratory measurements of growth and light attenuation for all strains, and run at two temperatures and light intensities. 17 000 runs were simulated for each pair using a statistical distribution with Monte Carlo approach. The model results showed that cyanobacterial competition was highly variable, depending on strains present, light and temperature conditions. There was no absolute 'winner' under all conditions as there were always strains predicted to coexist with the dominant strains, which were M. aeruginosa strains at 20°C and C. raciborskii strains at 28°C. The uncertainty in prediction of species competition outcomes was due to the substantial variability of growth responses within and between strains. Overall, this study demonstrates that within-species strain variability has a potentially large effect on cyanobacterial population dynamics, and therefore this variability may substantially reduce confidence in predicting outcomes of phytoplankton competition in deterministic models, that are based on only one set of parameters for each species or strain.

The efficiency of combined coagulant and ballast to remove harmful cyanobacterial blooms in a tropical shallow system.

We tested the hypothesis that a combination of coagulant and ballast could be efficient for removal of positively buoyant harmful cyanobacteria in shallow tropical waterbodies, and will not promote the release of cyanotoxins. This laboratory study examined the efficacy of coagulants [polyaluminium chloride (PAC) and chitosan (made of shrimp shells)] alone, and combined with ballast (lanthanum modified bentonite, red soil or gravel) to remove the natural populations of cyanobacteria collected from a shallow eutrophic urban reservoir with alternating blooms of Cylindrospermopsis and Microcystis. PAC combined with ballast was effective in settling blooms dominated by Microcystis or Cylindrospermopsis. Contrary to our expectation, chitosan combined with ballast was only effective in settling Cylindrospermopsis-dominated blooms at low pH, whereas at pH≥8 no effective flocculation and settling could be evoked. Chitosan also had a detrimental effect on Cylindrospermopsis causing the release of saxitoxins. In contrast, no detrimental effect on Microcystis was observed and all coagulant-ballast treatments were effective in not only settling the Microcystis dominated bloom, but also lowering dissolved microcystin concentrations. Our data show that the best procedure for biomass reduction also depends on the dominant species.

Massive fish mortality and Cylindrospermopsis raciborskii bloom in Aleksandrovac Lake.

This paper presents a case study of a massive fish mortality during a Cylindrospermopsis raciborskii bloom in Aleksandrovac Lake, Serbia in mid-December 2012. According to a preliminary investigation of the samples taken on November 6 before the fish mortalities and to extended analyses of samples taken on November 15, no values of significant physicochemical parameters emerged to explain the cause(s) of the fish mortality. No industrial pollutants were apparent at this location, and results excluded the likelihood of bacterial infections. Even after freezing, the dissolved oxygen concentration in the water was sufficient for fish survival. High concentrations of chlorophyll a and phaeophytin occurred in the lake, and phytoplankton bloom samples were lethal in Artemia salina bioassays. A bloom of the cyanobacterium C. raciborskii was recorded during November. Although the A. salina bioassays indicated the presence of toxic compounds in the cyanobacterial cells, the cyanotoxins, microcystins, cylindrospermopsin and saxitoxin were not detected.

Study of carotenoids in cyanobacteria by Raman spectroscopy.

Cyanobacteria have established dominant aquatic populations around the world, generally in aggressive environments and under severe stress conditions, e.g., intense solar radiation. Several marine strains make use of compounds such as the polyenic molecules for their damage protection justifying the range of colours observed for these species. The peridinin/chlorophyll-a/protein complex is an excellent example of essential structures used for self-prevention; their systems allow to them surviving under aggressive environments. In our simulations, few protective dyes are required to the initial specimen defense; this is an important data concern the synthetic priority in order to supply adequate damage protection. Raman measurements obtained with 1064 and 514.5 nm excitations for Cylindrospermopsis raciborskii and Microcystis aeruginosa strains shows bands assignable to the carotenoid peridinin. It was characterized by bands at 1940, 1650, 1515, 1449, 1185, 1155 and 1000 cm(-1) assigned to ν(C=C=C) (allenic vibration), ν(C=C/CO), ν(C=C), δ(C-H, C-18/19), δ(C-H), ν(C-C), and ρ(C-CH3), respectively. Recognition by Raman spectroscopy proved to be an important tool for preliminaries detections and characterization of polyene molecules in several algae, besides initiate an interesting discussion about their synthetic priority.

Microbial players involved in the decline of filamentous and colonial cyanobacterial blooms with a focus on fungal parasitism.

In the forthcoming decades, it is widely believed that the dominance of colonial and filamentous bloom-forming cyanobacteria (e.g. Microcystis, Planktothrix, Anabaena and Cylindrospermopsis) will increase in freshwater systems as a combined result of anthropogenic nutrient input into freshwater bodies and climate change. While the physicochemical parameters controlling bloom dynamics are well known, the role of biotic factors remains comparatively poorly studied. Morphology and toxicity often - but not always - limit the availability of cyanobacteria to filter feeding zooplankton (e.g. cladocerans). Filamentous and colonial cyanobacteria are widely regarded as trophic dead-ends mostly inedible for zooplankton, but substantial evidence shows that some grazers (e.g. copepods) can bypass this size constraint by breaking down filaments, making the bloom biomass available to other zooplankton species. A wide range of algicidal bacteria (mostly from the Alcaligenes, Flavobacterium/Cytophaga group and Pseudomonas) and viruses (Podoviridae, Siphoviridae and Myoviridae) may also contribute to bloom control, via their lytic activity underpinned by a diverse array of mechanisms. Fungal parasitism by the Chytridiomycota remains the least studied. While each of these biotic factors has traditionally been studied in isolation, emerging research consistently point to complex interwoven interactions between biotic and environmental factors.