Advancing knowledge on cyanobacterial blooms in freshwaters

Cyanobacterial blooms have become a frequent phenomenon in freshwaters worldwide; they are a widely known indicator of eutrophication and water quality deterioration. Information and knowledge contributing towards the evaluation of the ecological status of freshwaters, particularly since many are used for recreation, drinking water, and aquaculture, is valuable. This Special Issue, entitled "Advancing Knowledge on Cyanobacterial Blooms in Freshwaters", includes 11 research papers that will focus on the use of complementary approaches, from the most recently developed molecular-based methods to more classical approaches and experimental and mathematical modelling regarding the factors (abiotic and/or biotic) that control the diversity of not only the key bloom-forming cyanobacterial species, but also their interactions with other biota, either in freshwater systems or their adjacent habitats, and their role in preventing and/or promoting cyanobacterial growth and toxin production. © 2020 by the authors

Cyanotoxin contamination in commercial Spirulina food supplements

Arthrospira (“Spirulina”) is the most used cyanobacterium in food supplementation. “Spirulina” products are considered as superfood due to their excellent nutritional content, although several studies have shown the toxic effects of short and long term “Spirulina” consumption. In this work, a total of 31 “Spirulina” products from different brands, available in Greece, were investigated for their cyanotoxin contamination. All samples were positive for microcystins (MCYSTs) (35.7–583.5 ng g−1 dry weight [dw]). Two samples were tested positive for cylindrospermopsins (CYNs) (221.4–351.8 ng g−1 dw). No saxitoxins (STXs) were found. According to guidelines, the MCYSTs concentrations did not exceed the tolerable daily intake (TDI) set for adults, but six products exceeded the TDI set for children and 23 exceeded the TDI set for infants. The CYNs did not surpass the TDI for adults, but both samples exceeded the TDI for children and infants. This study showed that the low cyanobacterial toxin concentrations occurring in commercial “Spirulina” supplements, do not pose obvious risks for adults’ consumption, however they pose health risks for children and infants after daily consumption. To protect consumers, regular monitoring of these toxins at all stages of “Spirulina” production is recommended. © 2021, Bundesamt für Verbraucherschutz und Lebensmittelsicherheit (BVL)

Monitoring of cyanobacteria for water quality: doing the necessary right or wrong?

Cyanobacteria are an essential biological component of phytoplankton water quality assessment. However, there are some problems associated with the widely used everyday practices of sampling, estimation and use of cyanobacteria when calculating phytoplankton indices assessing water quality. Many indices were developed during the implementation of the Water Framework Directive, considered the most innovative European environmental legislation. Most indices include cyanobacteria as a composition or bloom metric. Problems with the indices concern the exclusion of most chroococcalean taxa from cyanobacterial biovolume estimations in lakes and reservoirs of the Mediterranean region, treatment of the mucilage of colonial chroococcalean taxa in biovolume estimations and overlooking of deep-water cyanobacterial blooms due to sampling depth. These problems may lead to a biased view of water quality. In this paper we argue in favour of including all cyanobacteria taxa and their mucilage in biovolume estimations and considering a sampling depth that covers deep-water maxima, such as those formed by Planktothrix rubescens or colonial chroococcalean taxa

Different phytoplankton descriptors show asynchronous changes in a shallow urban lake (L. Kastoria, Greece) after sewage diversion

Phytoplankton long-term changes were examined in a shallow urban lake (L. Kastoria, Greece) after sewage diversion in nine distinct years from 1994-2010. Using different levels of phytoplankton descriptors (diversity, taxonomic and functional composition, dominant species, total phytoplankton and cyanobacterial biomass) asynchronous changes were observed in the lake's phytoplankton community. The first dynamic response was observed five years after sewage diversion by a sharp decrease in total phytoplankton and cyanobacterial biomass. However, cyanobacteria remained the predominant taxonomic group. Only in 2005, a shift from cyanobacteria to chlorophytes-diatoms was observed, coinciding with a considerable decrease in the lake's water level due to flushing. A depletion of the long-lasting dominant filamentous cyanobacteria Limnothrix redekei and Cylindrospermopsis raciborskii was observed from 2005-2010. In contrast, a persistent dominance of toxic Microcystis species was recorded, indicating insufficient water quality. Fifteen years after sewage diversion, phytoplankton species richness doubled and functional diversity increased to include newly established species. Overall, the presented data provide insights into how the phytoplankton community assembles during restoration, with implications both for scientific analysis and ecological water quality management in the Mediterranean region

Molecular diversity of bacteria in commercially available "Spirulina" food supplements

The cyanobacterium Arthrospira is among the most well-known food supplements worldwide known as "Spirulina." While it is a widely recognized health-promoting natural product, there are no reports on the molecular diversity of commercially available brands of "Spirulina" supplements and the occurrence of other cyanobacterial and heterotrophic bacterial microorganisms in these products. In this study, 454- pyrosequencing analysis of the total bacterial occurrence in 31 brands of "Spirulina" dietary supplements from the Greek market was applied for the first time. In all samples, operational taxonomic units (OTUs) of Arthrospira platensis were the predominant cyanobacteria. Some products contained additional cyanobacterial OTUs including a few known potentially toxic taxa. Moreover, 469 OTUs were detected in all 31 products collectively, with most of them being related to the Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria and Verrucomicrobia. All samples included heterotrophic bacterial OTUs, ranging from 9-157 per product. Among the most common OTUs were ones closely related to taxa known for causing health issues (i.e., Pseudomonas, Flavobacterium, Vibrio, Aeromonas, Clostridium, Bacillus, Fusobacterium, Enterococcus). The observed high cyanobacterial and heterotrophic bacterial OTUs richness in the final product is a point for further research on the growth and processing of Arthrospira biomass for commercial purposes. © 2016 Vardaka et al

Morphological and molecular analysis of bloom-forming Cyanobacteria in two eutrophic, shallow Mediterranean lakes

We investigated the diversity of Cyanobacteria by microscopic observation and sequencing of cyanobacterial-specific amplified 16S rRNA genes in the water column of two shallow, eutrophic lakes (Doirani and Kastoria, northern Greece) during summer blooms. Previous phytoplankton studies in these lakes have shown that prolonged cyanobacterial blooms can occur, which are dominated by known toxic species, as well as other less known, co-occurring species. A total of 118 clones were sequenced which were grouped in 23 Cyanobacteria and 11 chloroplast-like phylotypes. Phylogenetic analysis revealed that each library included several unique phylotypes, as well as members of all common bloom-forming Cyanobacteria. Most of the phylotypes belonged to the genera Microcystis, Anabaena, Aphanizomenon, Cylindrospermopsis-Raphidiopsis group, Limnothrix and Planktothrix, comprising most of the diversity previously recognized by morphological observations in cyanobacterial morphospecies in these lakes. In addition, novel phylotypes belonging to the Chroococcales were recognized in both lakes. The structure of the cyanobacterial communities of the lakes were very similar, as revealed by 1 he diversity index H (2.06 and 2.01 for L Doirani and Kastoria, respectively) and LIBSHUFF analysis (XY(12) P-value = 0.122 and YX(12) P-value = 0.536), due to occurrence of groups of common phylotypes. This study gives an example for successful cyanobacterial bloom analysis by the combination of morphological and phylogenetic methods useful for monitoring cyanobacteria and water quality in freshwaters. (C) 2010 Elsevier GmbH. All rights reserved

Warming and Acidification Effects on Planktonic Heterotrophic Pico- and Nanoflagellates in a Mesocosm Experiment

We studied the response of the heterotrophic flagellate (HF) community to the combined impact of warming and ocean acidification in a mesocosm experiment with a plankton community from the western Baltic Sea. We performed a quantitative analysis of the response at the level of total biomass and size classes and a semi-quantitative one at the level of individual taxa. Total biomass of HF was significantly lower under higher temperatures while there was no significant effect of CO2. The mean biomass of the picoflagellates did not respond to temperature while the three nanoflagellate size classes (class limits 3, 5, 8, 15 μm) responded negatively to warming while not responding to CO2. The taxon-level results indicate that heterotrophic flagellates do not form a homogenous trophic guild, as often assumed in pelagic food web studies. Instead, the heterotrophic flagellates formed a “food web within the food web”. There was a pronounced succession of flagellates leading from a dominance of bacterivores and colloidal matter feeders before the phytoplankton bloom to omnivorous feeders preying upon phytoplankton and heterotrophic flagellates during and after the bloom. This complex intraguild predation patterns probably dampened the response to experimental treatments. © 2016 Elsevier Gmb

Raphidiopsis mediterranea Skuja represents non-heterocytous life-cycle stages of Cylindrospermopsis raciborskii (Woloszynska) Seenayya et Subba Raju in Lake Kastoria (Greece), its type locality: Evidence by morphological and phylogenetic analysis

The taxonomical relationship of Cylindrospermopsis raciborskii and Raphidiopsis mediterranea was studied by morphological and 16S rRNA gene diversity analyses of natural populations from Lake Kastoria, Greece. Samples were obtained during a bloom (23,830 trichomes mL(-1)) in August 2003. A high diversity of apical cell, trichome, heterocyte and akinete morphology, trichome fragmentation and reproduction was observed. Trichomes were grouped into three dominant morphotypes: the typical and the non-heterocytous morphotype of C. raciborskii and the typical morphotype of R. mediterranea. A morphometric comparison of the dominant morphotypes showed significant differences in mean values of cell and trichome sizes despite the high overlap in the range of the respective size values. Additionally, two new morphotypes representing developmental stages of the species are described while a new mode of reproduction involving a structurally distinct reproductive cell is described for the first time in planktic Nostocales. A putative life-cycle, common for C. raciborskii and R. mediterranea is proposed revealing that trichome reproduction of R. mediterranea gives rise both to R. mediterranea and C. raciborskii non-heterocytous morphotypes. The phylogenetic analysis of partial 16S rRNA gene (ca. 920 bp) of the co-existing Cylindrospermopsis and Raphidiopsis morphotypes revealed only one phylotype which showed 99.54% similarity to R. mediterranea HB2 (China) and 99.19% similarity to C. raciborskii form 1 (Australia). We propose that all morphotypes comprised stages of the life cycle of C. raciborkii whereas R. mediterranea from Lake Kastoria (its type locality) represents non-heterocytous stages of Cylindrospermopsis complex life cycle. (C) 2009 Elsevier B.V. All rights reserved

Molecular detection of potentially toxic cyanobacteria and their associated bacteria in lake water column and sediment

We investigated the molecular diversity of cyanobacteria and bacteria during a water bloom in a lake with a long history of toxic cyanobacterial blooms (Lake Kastoria, Greece). We also tested the hypothesis whether bloom-forming cyanobacteria are preserved in the lake's sediment 2 years after the bloom. The dominant cyanobacteria during the bloom included the potentially toxin-producing Microcystis aeruginosa and several other Chroococcales forms closely related to the genus Microcystis. This suggests that the use of cyanobacterial-specific primers seems to be very informative in describing the cyanobacteria during the water blooms. The bacterial community showed high diversity, consisting mostly of singleton and doubleton phylotypes. The majority of the phylotypes were typical lake bacteria including some potential pathogens and toxin metabolising bacteria, suggesting that the dominant toxic cyanobacteria did not have any significant effect on the bacterial community structure. In the sediment, 2 years after the water bloom, no bloom-forming cyanobacteria were retrieved, suggesting that they cannot be preserved in the sediment. Similar to the water column, sediment bacterial diversity was also high, consisting mostly of yet-uncultured bacteria that are related to environments where organic matter degradation takes place

Haematococcus: A successful air-dispersed colonist in ephemeral waters is rarely found in phytoplankton communities

In a literature search, the presence of Haematococcus in phytoplankton communities and its biogeography were investigated. Haematococcus, although showing a wide biogeographical distribution, has been rarely found in phytoplankton communities. Simultaneously, the colonization potential of air-dispersed Haematococcus in ephemeral waters and its interactions with coexisting phytoplankton taxa were examined by microscopy and molecular methods. Haematococcus was a successful colonist, appearing among the first taxa in the experimental containers. According to principal component analysis, Haematococcus growth rate was negatively correlated with the abundance and species richness of the other autotrophs. Furthermore, a negative correlation between Haematococcus and Chlamydomonas and a positive one between Haematococcus and Chlorella were found. Overall, Haematococcus appears to be an effective air-dispersed alga that can successfully colonize and establish populations in small ephemeral water bodies. However, its absence from phytoplankton in larger permanent water bodies could be related to its high light requirements, its competitive disadvantages against other algae, and the grazing pressures from predators. The results of our study suggest a life strategy based on adaptation to higher light intensities in very shallow waters compared with optical dense lakes. Therefore, ephemeral waters are the regular habitat for Haematococcus instead of being “stepping stones” for the colonization of lake phytoplankton. © TÜBİTAK