Marine Microbiota Responses to Shipping Scrubber Effluent Assessed at Community Structure and Function Endpoints.

The use of novel high-throughput sequencing (HTS) technologies to examine the responses of natural multidomain microbial communities to scrubber effluent discharges to the marine environment is still limited. Thus, we applied metabarcoding sequencing targeting the planktonic unicellular eukaryotic and prokaryotic fraction (phytoplankton, bacterioplankton, and protozooplankton) in mesocosm experiments with natural microbial communities from a polluted and an unpolluted site. Furthermore, metagenomic analysis revealed changes in the taxonomic and functional dominance of multidomain marine microbial communities after scrubber effluent additions. The results indicated a clear shift in the microbial communities after such additions, which favored bacterial taxa with known oil and polycyclic aromatic hydrocarbons (PAHs) biodegradation capacities. These bacteria exhibited high connectedness with planktonic unicellular eukaryotes employing variable trophic strategies, suggesting that environmentally relevant bacteria can influence eukaryotic community structure. Furthermore, Clusters of Orthologous Genes associated with pathways of PAHs and monocyclic hydrocarbon degradation increased in numbers at treatments with high scrubber effluent additions acutely. These genes are known to express enzymes acting at various substrates including PAHs. These indications, in combination with the abrupt decrease in the most abundant PAHs in the scrubber effluent below the limit of detection-much faster than their known half-lives-could point toward a bacterioplankton-initiated rapid ultimate biodegradation of the most abundant toxic contaminants of the scrubber effluent. The implementation of HTS could be a valuable tool to develop multilevel biodiversity indicators of the scrubber effluent impacts on the marine environment, which could lead to improved impact assessment. Environ Toxicol Chem 2024;43:1012-1029. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

A semi-continuous algal-bacterial wastewater treatment process coupled with bioethanol production.

Harnessing the biomass energy potential through biofuel production offers new outlets for a circular economy. In this study an integrated system which combine brewery wastewater treatment using algal-bacterial aggregates instead of activated sludge was developed. The use of algal-bacterial aggregates can eliminate the aeration requirements and significantly reduce the high biomass harvesting costs associated with algal monocultures. A sequencing batch reactor (SBR) setup operating with and without biomass recirculation was used to investigate pollutant removal rates, aggregation capacity and microbial community characteristics under a range of hydraulic retention times (HRTs) and solid retention times (SRTs). It was observed that biomass recirculation strategy significantly enhanced aggregation and pollutant removal (i.e., 78.7%, 94.2% and 75.2% for d-COD, TKN, and PO43--P, respectively). The microbial community established was highly diverse consisting of 161 Bacterial Operational Taxonomic Units (B-OTUs) and 16 unicellular Eukaryotic OTUs (E-OTUs). Escalation the optimal conditions (i.e., HRT = 4 d, SRT = 10 d) at pilot-scale resulted in nutrient starvation leading to 38-44% w/w carbohydrate accumulation. The harvested biomass was converted to bioethanol after acid hydrolysis followed by fermentation with Saccharomyces cerevisiae achieving a bioethanol production yield of 0.076 g bioethanol/g biomass. These data suggest that bioethanol production coupled with high-performance wastewater treatment using algal-bacterial aggregates is feasible, albeit less productive concerning bioethanol yields than systems exclusively designed for third and fourth-generation biofuel production.

A catastrophic change in a european protected wetland: From harmful phytoplankton blooms to fish and bird kill.

Understanding the processes that underlay an ecological disaster represents a major scientific challenge. Here, we investigated phytoplankton and zooplankton community changes before and during a fauna mass kill in a European protected wetland. Evidence on gradual development and collapse of harmful phytoplankton blooms, allowed us to delineate the biotic and abiotic interactions that led to this ecological disaster. Before the mass fauna kill, mixed blooms of known harmful cyanobacteria and the killer alga Prymnesium parvum altered biomass flow and minimized zooplankton resource use efficiency. These blooms collapsed under high nutrient concentrations and inhibitory ammonia levels, with low phytoplankton biomass leading to a dramatic drop in photosynthetic oxygenation and a shift to a heterotrophic ecosystem phase. Along with the phytoplankton collapse, extremely high numbers of red planktonic crustaceans-Daphnia magna, visible through satellite images, indicated low oxygen conditions as well as a decrease or absence of fish predation pressure. Our findings provide clear evidence that the mass episode of fish and birds kill resulted through severe changes in phytoplankton and zooplankton dynamics, and the alternation on key abiotic conditions. Our study highlights that plankton-related ecosystem functions mirror the accumulated heavy anthropogenic impacts on freshwaters and could reflect a failure in conservation and restoration measures.

Air-dispersed aquatic microorganisms show establishment and growth preferences in different freshwater colonisation habitats.

We attempted to mimic aeolian ecosystems to examine how filters posed by regional characteristics can influence the establishment and growth of airborne microcolonisers of a common air source. Using a natural single source of aerosols we applied a combined microscopy and high-throughput sequencing approach to examine the diversity, settling and growth potential of air-dispersed microbes in water containers representing newly formed aquatic colonisation habitats of different trophic states and salinity. Heterotrophic microeukaryotes were favoured as initial settlers when nutrients were low, while autotrophs rapidly proliferated in the high-nutrient containers, possibly due to favourable germinating conditions for their preferred mode of dispersal with resting spores. Following settling of colonisers, we investigated two contrasting hypotheses: if the different water colonisation habitats harboured the same microbial communities after establishment and growth periods, this would point towards a selection of best-fit cosmopolitan colonisers, regardless of habitat-specific characteristics. Alternatively, community dissimilarities after the growth period would suggest a selection of settlers due to bottom-up controls combined with priority effects. Both analyses suggested that the structure of the microbial communities in the different colonisation habitats were driven by nutrient content and salinity, showing clustering to similar bottom-up forces and dissimilarities in significantly different colonisation habitats.

Implementation of the Water Framework Directive: Lessons Learned and Future Perspectives for an Ecologically Meaningful Classification Based on Phytoplankton of the Status of Greek Lakes, Mediterranean Region.

The enactment of the Water Framework Directive (WFD) initiated scientific efforts to develop reliable methods for comparing prevailing lake conditions against reference (or nonimpaired) states, using the state of a set biological elements. Drawing a distinction between impaired and natural conditions can be a challenging exercise. Another important aspect is to ensure that water quality assessment is comparable among the different Member States. In this context, the present paper offers a constructive critique of the practices followed during the WFD implementation in Greece by pinpointing methodological weaknesses and knowledge gaps that undermine our ability to classify the ecological quality of Greek lakes. One of the pillars of WDF is a valid lake typology that sets ecological standards transcending geographic regions and national boundaries. The national typology of Greek lakes has failed to take into account essential components. WFD compliance assessments based on the descriptions of phytoplankton communities are oversimplified and as such should be revisited. Exclusion of most chroococcal species from the analysis of cyanobacteria biovolume in Greek lakes/reservoirs and most reservoirs in Spain, Portugal, and Cyprus is not consistent with the distribution of those taxa in lakes. Similarly, the total biovolume reference values and the indices used in classification schemes reflect misunderstandings of WFD core principles. This hampers the comparability of ecological status across Europe and leads to quality standards that are too relaxed to provide an efficient target for the protection of Greek/transboundary lakes such as the ancient Lake Megali Prespa.

Response of a coastal Baltic Sea diatom-dominated phytoplankton community to experimental heat shock and changing salinity.

Climate change has been altering the ocean environment, affecting as a consequence the biological communities including microorganisms. We performed a mesocosm experiment to test whether biodiversity loss caused by one stressor would influence plankton community sensitivity to a subsequent stressor, as envisioned in Vinebrooke's multiple stressor concept. A natural Baltic Sea diatom-dominated phytoplankton assemblage was used as a model system where we examined whether a preceding heat shock would affect the community's response to changing salinity. Initially, the community was treated by a short-term temperature increase of 6 °C, which resulted in a loss of species compared to the control. Thereafter, the control and the heat-shocked communities were subject to a salinity change (- 5 psu, control, + 5 psu). The species Skeletonema dohrnii, Thalassiosira anguste-lineata, Thalassiosira nordenskioeldii, Chaetoceros socialis and Ditylum brightwellii were major components of the control and heat-shocked assemblages (> 80% of the total biomass). We examined the effect on species composition and biodiversity (morphospecies and operational taxonomic units (OTUs) related to phytoplankton) and on phytoplankton biomass. In addition, we explored the single species response of five dominant diatoms on these environmental perturbations. Our results showed that increased salinity significantly reduced the OTUs richness both in the control and the less diverse heated community as well as the phytoplankton biomass in the heated community. On the other hand, decreased salinity significantly increased species richness and phytoplankton biomass in both communities and OTUs richness in the control community. The five dominant diatoms reached their highest biomass under decreased salinity and responded negatively to increased salinity (lower biomass than ambient salinity). Contrary to Vinebrooke's multiple stressor concept, there was no indication that the heat treatment had altered the community's sensitivity to the salinity stress in our study system.

Occurrence and diversity of cyanotoxins in Greek lakes.

Toxic cyanobacteria occur in Greek surface water bodies. However, studies on the occurrence of cyanotoxins (CTs) are often limited to mainly microcystins (MCs), with use of screening methods, such as ELISA, that are not conclusive of the chemical structure of the CT variants and can be subject to false positive results. A multi-lake survey in Greece (14 lakes) was conducted in water and biomass, targeted to a wide range of multi-class CTs including MCs, nodularin-R (NOD), cylindrospermopsin (CYN), anatoxin-a (ANA-a) and saxitoxins (STXs), using multi-class/variant LC-MS/MS analytical workflows, achieving sensitive detection, definitive identification and accurate quantitation. A wide variety of CTs (CYN, ANA-a, STX, neoSTX, dmMC-RR, MC-RR, MC-YR, MC-HtyR, dm3MC-LR, MC-LR, MC-HilR, MC-WR, MC-LA, MC-LY, MC-LW and MC-LF), were detected, with MCs being the most commonly occurring. In biomass, MC-RR was the most abundant toxin, reaching 754 ng mg-1 dw, followed by MC-LR (458 ng mg-1 dw). CYN and ANA-a were detected for the first time in the biomass of Greek lakes at low concentrations and STXs in lakes Trichonis, Vistonis and Petron. The abundance and diversity of CTs were also evaluated in relation to recreational health risks, in a case study with a proven history of MCs (Lake Kastoria).

Unicellular Eukaryotic Community Response to Temperature and Salinity Variation in Mesocosm Experiments.

Climate change has profound impacts on marine biodiversity and biodiversity changes in turn might affect the community sensitivity to impacts of abiotic changes. We used mesocosm experiments and Next Generation Sequencing to study the response of the natural Baltic and Mediterranean unicellular eukaryotic plankton communities (control and +6°C heat shock) to subsequent salinity changes (-5 psu, +5 psu). The impact on Operational Taxonomic Unit (OTU) richness, taxonomic and functional composition and rRNA:rDNA ratios were examined. Our results showed that heat shock leads to lower OTU richness (21% fewer OTUs in the Baltic and 14% fewer in the Mediterranean) and a shift in composition toward pico- and nanophytoplankton and heterotrophic related OTUs. Heat shock also leads to increased rRNA:rDNA ratios for pico- and micrograzers. Less than 18% of shared OTUs were found among the different salinities indicating the crucial role of salinity in shaping communities. The response of rRNA:rDNA ratios varied highly after salinity changes. In both experiments the diversity decrease brought about by heat shock influenced the sensitivity to salinity changes. The heat shock either decreased or increased the sensitivity of the remaining community, depending on whether it removed the more salinity-sensitive or the salinity-tolerant taxa.

Resisting annihilation: relationships between functional trait dissimilarity, assemblage competitive power and allelopathy.

Allelopathic species can alter biodiversity. Using simulated assemblages that are characterised by neutrality, lumpy coexistence and intransitivity, we explore relationships between within-assemblage competitive dissimilarities and resistance to allelopathic species. An emergent behaviour from our models is that assemblages are more resistant to allelopathy when members strongly compete exploitatively (high competitive power). We found that neutral assemblages were the most vulnerable to allelopathic species, followed by lumpy and then by intransitive assemblages. We find support for our modeling in real-world time-series data from eight lakes of varied morphometry and trophic state. Our analysis of this data shows that a lake's history of allelopathic phytoplankton species biovolume density and dominance is related to the number of species clusters occurring in the plankton assemblages of those lakes, an emergent trend similar to that of our modeling. We suggest that an assemblage's competitive power determines its allelopathy resistance.

A Leptolyngbya-based microbial consortium for agro-industrial wastewaters treatment and biodiesel production.

A mixed cyanobacterial-mixotrophic algal population, dominated by the filamentous cyanobacterium Leptolyngbya sp. and the microalga Ochromonas (which contributed to the total photosynthetic population with rates of less than 5%), was studied under non-aseptic conditions for its efficiency to remove organic and inorganic compounds from different types of wastes/wastewaters while simultaneously producing lipids. Second cheese whey, poplar sawdust, and grass hydrolysates were used in lab-scale experiments, in photobioreactors that operated under aerobic conditions with different initial nutrient (C, N and P) concentrations. Nutrient removal rates, biomass productivity, and the maximum oil production rates were determined. The highest lipid production was achieved using the biologically treated dairy effluent (up to 14.8% oil in dry biomass corresponding to 124 mg L-1) which also led to high nutrient removal rates (up to 94%). Lipids synthesized by the microbial consortium contained high percentages of saturated and mono-unsaturated fatty acids (up to 75% in total lipids) for all the substrates tested, which implies that the produced biomass may be harnessed as a source of biodiesel.

Do marine phytoplankton follow Bergmann's rule sensu lato?

Global warming has revitalized interest in the relationship between body size and temperature, proposed by Bergmann's rule 150 years ago, one of the oldest manifestations of a 'biogeography of traits'. We review biogeographic evidence, results from clonal cultures and recent micro- and mesocosm experiments with naturally mixed phytoplankton communities regarding the response of phytoplankton body size to temperature, either as a single factor or in combination with other factors such as grazing, nutrient limitation, and ocean acidification. Where possible, we also focus on the comparison between intraspecific size shifts and size shifts resulting from changes in species composition. Taken together, biogeographic evidence, community-level experiments and single-species experiments indicate that phytoplankton average cell sizes tend to become smaller in warmer waters, although temperature is not necessarily the proximate environmental factor driving size shifts. Indirect effects via nutrient supply and grazing are important and often dominate. In a substantial proportion of field studies, resource availability is seen as the only factor of relevance. Interspecific size effects are greater than intraspecific effects. Direct temperature effects tend to be exacerbated by indirect ones, if warming leads to intensified nutrient limitation or copepod grazing while ocean acidification tends to counteract the temperature effect on cell size in non-calcifying phytoplankton. We discuss the implications of the temperature-related size trends in a global-warming context, based on known functional traits associated with phytoplankton size. These are a higher affinity for nutrients of smaller cells, highest maximal growth rates of moderately small phytoplankton (ca. 102 µm3 ), size-related sensitivities for different types of grazers, and impacts on sinking rates. For a phytoplankton community increasingly dominated by smaller algae we predict that: (i) a higher proportion of primary production will be respired within the microbial food web; (ii) a smaller share of primary production will be channeled to the classic phytoplankton - crustacean zooplankton - fish food chain, thus leading to decreased ecological efficiency from a fish-production point of view; (iii) a smaller share of primary production will be exported through sedimentation, thus leading to decreased efficiency of the biological carbon pump.

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, 15µm) 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.

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.

Warming and Ocean Acidification Effects on Phytoplankton--From Species Shifts to Size Shifts within Species in a Mesocosm Experiment.

While the isolated responses of marine phytoplankton to climate warming and to ocean acidification have been studied intensively, studies on the combined effect of both aspects of Global Change are still scarce. Therefore, we performed a mesocosm experiment with a factorial combination of temperature (9 and 15 °C) and pCO2 (means: 439 ppm and 1040 ppm) with a natural autumn plankton community from the western Baltic Sea. Temporal trajectories of total biomass and of the biomass of the most important higher taxa followed similar patterns in all treatments. When averaging over the entire time course, phytoplankton biomass decreased with warming and increased with CO2 under warm conditions. The contribution of the two dominant higher phytoplankton taxa (diatoms and cryptophytes) and of the 4 most important species (3 diatoms, 1 cryptophyte) did not respond to the experimental treatments. Taxonomic composition of phytoplankton showed only responses at the level of subdominant and rare species. Phytoplankton cell sizes increased with CO2 addition and decreased with warming. Both effects were stronger for larger species. Warming effects were stronger than CO2 effects and tended to counteract each other. Phytoplankton communities without calcifying species and exposed to short-term variation of CO2 seem to be rather resistant to ocean acidification.

Spatially uniform but temporally variable bacterioplankton in a semi-enclosed coastal area.

Studies focusing on the temporal and spatial dynamics of bacterioplankton communities within littoral areas undergoing direct influences from the coast are quite limited. In addition, they are more complicated to resolve compared to communities in the open ocean. In order to elucidate the effects of spatial vs. temporal variability on bacterial communities in a highly land-influenced semi-enclosed gulf, surface bacterioplankton communities from five coastal sites in Igoumenitsa Gulf (Ionian Sea, Greece) were analyzed over a nine-month period using 16S rDNA 454-pyrosequencing. Temporal differences were more pronounced than spatial ones, with lower diversity indices observed during the summer months. During winter and early spring, bacterial communities were dominated by SAR11 representatives, while this pattern changed in May when they were abruptly replaced by members of Flavobacteriales, Pseudomonadales, and Alteromonadales. Additionally, correlation analysis showed high negative correlations between the presence of SAR11 OTUs in relation to temperature and sunlight that might have driven, directly or indirectly, the disappearance of these OTUs in the summer months. The dominance of SAR11 during the winter months further supported the global distribution of the clade, not only in the open-sea, but also in coastal systems. This study revealed that specific bacteria exhibited distinct succession patterns in an anthropogenic-impacted coastal system. The major bacterioplankton component was represented by commonly found marine bacteria exhibiting seasonal dynamics, while freshwater and terrestrial-related phylotypes were absent.

Molecular diversity reveals previously undetected air-dispersed protist colonists in a Mediterranean area.

The molecular diversity of air-dispersed protists was examined through the 18S rRNA gene clone library construction in air samples and samples from experimental water containers passively collecting air-dispersed microorganisms, from July 2007 till October 2008 in three different sites of Northern Greece. The majority of the samplings took place in an urban industrialized coastal city (Thessaloniki). In all the samples, a total of 29 unique phylotypes were detected belonging to 10 known major taxonomic groups. The most abundant phylotypes were affiliated to known taxa of Ciliophora and Chlorophyceae, commonly found in various habitats. Additionally, various previously unnoticed and under-studied taxa, such as Bicosoecida, Oomycetes and Labyrinthulomycetes, were detected. These taxa are potentially important in ecological processes, through dispersal and colonization of various habitats. Multivariate statistical analysis associated the most abundant phylotypes with rainfall, suggesting that rain is a favorable means for reposition of air-dispersed protists. This is the first study investigating the molecular diversity of air-dispersed protists, including algae and heterotrophic protists.

Plankton microorganisms coinciding with two consecutive mass fish kills in a newly reconstructed lake.

Lake Karla, Greece, was dried up in 1962 and its refilling started in 2009. We examined the Cyanobacteria and unicellular eukaryotes found during two fish kill incidents, in March and April 2010, in order to detect possible causative agents. Both microscopic and molecular (16S/18S rRNA gene diversity) identification were applied. Potentially toxic Cyanobacteria included representatives of the Planktothrix and Anabaena groups. Known toxic eukaryotes or parasites related to fish kill events were Prymnesium parvum and Pfiesteria cf. piscicida, the latter being reported in an inland lake for the second time. Other potentially harmful microorganisms, for fish and other aquatic life, included representatives of Fungi, Mesomycetozoa, Alveolata, and Heterokontophyta (stramenopiles). In addition, Euglenophyta, Chlorophyta, and diatoms were represented by species indicative of hypertrophic conditions. The pioneers of L. Karla's plankton during the first months of its water refilling process included species that could cause the two observed fish kill events.

Phytoplankton and water quality in a Mediterranean drinking-water reservoir (Marathonas Reservoir, Greece).

Phytoplankton and water quality of Marathonas drinking-water Reservoir were examined for the first time. During the study period (July-September 2007), phytoplankton composition was indicative of eutrophic conditions although phytoplankton biovolume was low (max. 2.7 mm³ l⁻¹). Phytoplankton was dominated by cyanobacteria and diatoms, whereas desmids and dinoflagellates contributed with lower biovolume values. Changing flushing rate in the reservoir (up to 0.7% of reservoir's water volume per day) driven by water withdrawal and occurring in pulses for a period of 15-25 days was associated with phytoplankton dynamics. Under flushing pulses: (1) biovolume was low and (2) both 'good' quality species and the tolerant to flushing 'nuisance' cyanobacterium Microcystis aeruginosa dominated. According to the Water Framework Directive, the metrics of phytoplankton biovolume and cyanobacterial percentage (%) contribution indicated a moderate ecological water quality. In addition, the total biovolume of cyanobacteria as well as the dominance of the known toxin-producing M. aeruginosa in the reservoir's phytoplankton indicated a potential hazard for human health according to the World Health Organization.

Airborne algae and cyanobacteria: occurrence and related health effects.

Published information on airborne algae and cyanobacteria worldwide and the related human health effects is scarce. Since 1844, a total of 353 morphological taxa (genera or species) have been identified in aerobiological studies. However, due to diverse methodologies and different microorganisms targeted in these studies, direct comparisons on the occurrences of airborne algae and cyanobacteria in various studies are rather dubious. Thirty-eight airborne algae and cyanobacteria were shown to induce allergy, skin irritation, hay fever, rhinitis, sclerosis and respiratory problems when aerosolized and inhaled. Another 14 airborne taxa are known toxin producers posing threat to human health. Most frequently associated with health effects are the genera Chlorella, Scenedesmus, Chlorococcum, Klebsormidium (Hormidium) and Lyngbya. In the air of the Mediterranean city of Thessaloniki, we found 63 algal and cyanobacterial taxa, with 21 reported for the first time in the air. Seven taxa were potentially harmful. Algae and cyanobacteria can contribute significantly in the total air particle load, rendering them as causative agents for health issues when inhaled.

Diversity of cyanobacterial phylotypes in a Mediterranean drinking water reservoir (Marathonas, Greece).

The structure of the cyanobacterial community in a large drinking water reservoir (Marathonas, Greece) was investigated in October 2007 and September 2008. Cyanobacteria-specific primers were used for the PCR amplification of cyanobacterial 16S rDNAs from three water column sites and the water collection tank. In total, 199 clones were sequenced representing 52 unique cyanobacterial, including chloroplast-related, and 11 non-cyanobacterial phylotypes. All cyanobacterial phylotypes belonged to the order Chroococcales. Cluster analysis showed that the cyanobacterial communities in 2007 in the three water column sites showed high similarity between the stations and low diversity (H=1.17-1.44), due to the occurring common phylotypes, while all sites in 2008 had very low similarities between them and higher diversity (H=1.56-2.40). Some of the most abundant phylotypes were closely related (>98%) to members of the genus Gloeocapsa and a potentially toxin-producing strain of Microcystis aeruginosa. The non-cyanobacterial phylotypes were either unaffiliated or belonged to the Verrucomicrobia, and were related with sequences originating from lake water habitats.