Hydantoanabaenopeptins from Lake Kinneret Microcystis Bloom, Isolation, and Structure Elucidation of the Possible Intermediates in the Anabaenopeptins Biosynthesis.

Anabaenopeptins are common metabolites of cyanobacteria. In the course of reisolation of the known aeruginosins KT608A and KT608B for bioassay studies, we noticed the presence of some unknown anabaenopeptins in the extract of a Microcystis cell mass collected during the 2016 spring bloom event in Lake Kinneret, Israel. The 1H NMR spectra of some of these compounds presented a significant difference in the appearance of the ureido bridge protons, and their molecular masses did not match any one of the 152 known anabaenopeptins. Analyses of the 1D and 2D NMR, HRMS, and MS/MS spectra of the new compounds revealed their structures as the hydantoin derivatives of anabaenopeptins A, B, F, and 1[Dht]-anabaenopeptin A and oscillamide Y (1, 2, 3, 6, and 4, respectively) and a new anabaenopeptin, 1[Dht]-anabaenopeptin A (5). The known anabaenopeptins A, B, and F and oscillamide Y (7, 8, 9, and 10, respectively) were present in the extract as well. We propose that 1-4 and 6 are the possible missing intermediates in the previously proposed partial biosynthesis route to the anabaenopeptins. Compounds 1-6 were tested for inhibition of the serine proteases trypsin and chymotrypsin and found inactive at a final concentration of ca. 54 µM.

Gloeotrichia pisum in Lake Kinneret: A successful epiphytic cyanobacterium.

With climate change and re-oligotrophication of lakes due to restoration efforts, the relative importance of benthic cyanobacteria is increasing, but they are much less studied than their planktonic counterparts. Following a major water level rise event that inundated massive reed stands in Lake Kinneret, Israel, we discovered the appearance of a vast abundance of Gloeotrichia pisum (cyanobacteria). This provided an opportunity to investigate the biology and ecology of a benthic epiphytic colonial cyanobacterium, proliferating under altered environmental conditions, with possible toxin production potential and as a model for an invasive epiphyte. The species was identified by its typical morphology, and by sequencing its 16S rRNA gene and the intragenic space. We report on the abundance and spatial distribution of the detected colonies, their morphological characteristics, and pigment composition. High phycoerythrin content provides a brownish color and supports growth at low light levels. Genomic community composition analysis revealed that G. pisum colonies host a diverse microbial community of microalgae, cyanobacteria, bacteria, and archaea with a conserved and characteristic taxonomic composition. The Synechococcales order showed high relative abundance in the colony, as well as other prokaryotes producing secondary metabolites, such as the rhodopsin producer Pseudorhodobacter. The microbial consortium in the colonies performed nitrogen fixation. The diazotroph's phylogenetic relations were demonstrated. Tests for the presence of cyanotoxins (microcystin and cylindrospermopsin) proved negative. This study is the first documentation of this genus in Israel, providing insights into the invasive nature of G. pisum and the ecological implications of its appearance in a lake ecosystem.

Freshwater microbial metagenomes sampled across different water body characteristics, space and time in Israel.

Freshwater bodies are critical components of terrestrial ecosystems. The microbial communities of freshwater ecosystems are intimately linked water quality. These microbes interact with, utilize and recycle inorganic elements and organic matter. Here, we present three metagenomic sequence datasets (total of 182.9 Gbp) from different freshwater environments in Israel. The first dataset is from diverse freshwater bodies intended for different usages - a nature reserve, irrigation and aquaculture facilities, a tertiary wastewater treatment plant and a desert rainfall reservoir. The second represents a two-year time-series, collected during 2013-2014 at roughly monthly intervals, from a water reservoir connected to an aquaculture facility. The third is from several time-points during the winter and spring of 2015 in Lake Kinneret, including a bloom of the cyanobacterium Microcystis sp. These datasets are accompanied by physical, chemical, and biological measurements at each sampling point. We expect that these metagenomes will facilitate a wide range of comparative studies that seek to illuminate new aspects of freshwater microbial ecosystems and inform future water quality management approaches.

Dissolved organic nitrogen, dinoflagellates and cyanobacteria in two eutrophic lakes: Analysis by inferential modelling.

The Lake Suwa (Japan) has a history of non-N-fixing Microcystis blooms. Lake Kinneret (Israel) experienced multiannual periods of sole domination by the dinoflagellate Peridinium gatunense and periods dominated seasonally by P. gatunense or cyanobacteria. Extensive studies have been carried out in both lakes regarding the role of dissolved inorganic nitrogen and phosphorus as drivers of primary productivity. There is growing evidence that dissolved organic nitrogen (DON) compounds also influence not only biomass and structure of phytoplankton communities but also microcystin production. This study focuses on relationships of DON with: (1) population dynamics of Microcystis spp. and concentrations of microcystins in Lake Suwa, and (2) population dynamics of P. gatunense as well as N- and non-N-fixing cyanobacteria in Lake Kinneret. Modelling results for historical data of Lake Suwa by means of the hybrid evolutionary algorithm HEA revealed that the prediction of abundances of four Microcystis species and concentrations of cyanotoxins achieved higher coefficients of correlation when DON/DIN-ratios were included as drivers. Population dynamics of P. gatunense in Lake Kinneret appeared to have a strong inverse relationships with DON/DIN-ratios reflected by inferential models of HEA with higher coefficients of correlation when driven by DON/DIN-ratios. When DON/DIN-ratios were included as drivers, models of Microcystis spp. in Lake Kinneret performed higher coefficients of determination compared to models of N-fixing cyanobacteria. The study highlights the need to consider DON for improved understanding and management of population dynamics of cyanobacteria and dinoflagellates in freshwater lakes.

Cyanobacterial pigment concentrations in inland waters: Novel semi-analytical algorithms for multi- and hyperspectral remote sensing data.

Cyanobacteria are notorious for producing harmful algal blooms that present an ever-increasing serious threat to aquatic ecosystems worldwide, impacting the quality of drinking water and disrupting the recreational use of many water bodies. Remote sensing techniques for the detection and quantification of cyanobacterial blooms are required to monitor their initiation and spatiotemporal variability. In this study, we developed a novel semi-analytical approach to estimate the concentration of cyanobacteria-specific pigment phycocyanin (PC) and common phytoplankton pigment chlorophyll a (Chl a) from hyperspectral remote sensing data. The PC algorithm was derived from absorbance-concentration relationship, and the Chl a algorithm was devised based on a conceptual three-band structure model. The developed algorithms were applied to satellite imageries obtained by the Hyperspectral Imager for the Coastal Ocean (HICO™) sensor and tested in Lake Kinneret (Israel) during strong cyanobacterium Microcystis sp. bloom and out-of-bloom times. The sensitivity of the algorithms to errors was evaluated. The Chl a and PC concentrations were estimated with a mean absolute percentage difference (MAPD) of 16% and 28%, respectively. Sensitivity analysis shows that the influences of backscattering and other water constituents do not affect the estimation accuracy of PC (~2% MAPD). The reliable PC/Chl a ratios can be obtained at PC concentrations above 10 mg m-3. The computed PC/Chl a ratio depicts the contribution of cyanobacteria to the total phytoplankton biomass and permits investigating the role of ambient factors in the formation of a complex planktonic community. The novel algorithms have extensive practical applicability and should be suitable for the quantification of PC and Chl a in aquatic ecosystems using hyperspectral remote sensing data as well as data from future multispectral remote sensing satellites, if the respective bands are featured in the sensor.

Can Alkyl Quaternary Ammonium Cations Substitute H2O2 in Controlling Cyanobacterial Blooms-Laboratory and Mesocosm Studies.

Mitigation of harmful cyanobacterial blooms that constitute a serious threat to water quality, particularly in eutrophic water, such as in aquaculture, is essential. Thus, in this study, we tested the efficacy of selected cyanocides towards bloom control in laboratory and outdoor mesocosm experiments. Specifically, we focused on the applicability of a group of cationic disinfectants, alkyltrimethyl ammonium (ATMA) compounds and H2O2. The biocidal effect of four ATMA cations with different alkyl chain lengths was evaluated ex situ using Microcystis colonies collected from a fish pond. The most effective compound, octadecyl trimethyl ammonium (ODTMA), was further evaluated for its selectivity towards 24 cyanobacteria and eukaryotic algae species, including Cyanobacteria, Chlorophyta, Bacillariophyta, Euglenozoa and Cryptophyta. The results indicated selective inhibition of cyanobacteria by ODTMA-Br (C18) on both Chroccocales and Nostocales, but a minor effect on Chlorophytes and Bacillariophytes. The efficacy of ODTMA-Br (C18) (6.4 µM) in mitigating the Microcystis population was compared with that of a single low dose of H2O2 treatments (117.6 µM). ODTMA-Br (C18) suppressed the regrowth of Microcystis for a longer duration than did H2O2. The results suggested that ODTMA-Br (C18) may be used as an effective cyanocide and that it is worth further evaluating this group of cationic compounds as a treatment to mitigate cyanobacterial blooms in aquaculture.

Cyanobacterial Harmful Algal Blooms in Aquatic Ecosystems: A Comprehensive Outlook on Current and Emerging Mitigation and Control Approaches.

An intensification of toxic cyanobacteria blooms has occurred over the last three decades, severely affecting coastal and lake water quality in many parts of the world. Extensive research is being conducted in an attempt to gain a better understanding of the driving forces that alter the ecological balance in water bodies and of the biological role of the secondary metabolites, toxins included, produced by the cyanobacteria. In the long-term, such knowledge may help to develop the needed procedures to restore the phytoplankton community to the pre-toxic blooms era. In the short-term, the mission of the scientific community is to develop novel approaches to mitigate the blooms and thereby restore the ability of affected communities to enjoy coastal and lake waters. Here, we critically review some of the recently proposed, currently leading, and potentially emerging mitigation approaches in-lake novel methodologies and applications relevant to drinking-water treatment.

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.

Seasonal Dynamics Are the Major Driver of Microbial Diversity and Composition in Intensive Freshwater Aquaculture.

Aquaculture facilities such as fishponds are one of the most anthropogenically impacted freshwater ecosystems. The high fish biomass reared in aquaculture is associated with an intensive input into the water of fish-feed and fish excrements. This nutrients load may affect the microbial community in the water, which in turn can impact the fish health. To determine to what extent aquaculture practices and natural seasonal cycles affect the microbial populations, we characterized the microbiome of an inter-connected aquaculture system at monthly resolution, over 3 years. The system comprised two fishponds, where fish are grown, and an operational water reservoir in which fish are not actively stocked. Clear natural seasonal cycles of temperature and inorganic nutrients concentration, as well as recurring cyanobacterial blooms during summer, were observed in both the fishponds and the reservoir. The structure of the aquatic bacterial communities in the system, characterized using 16S rRNA sequencing, was explained primarily by the natural seasonality, whereas aquaculture-related parameters had only a minor explanatory power. However, the cyanobacterial blooms were characterized by different cyanobacterial clades dominating at each fishpond, possibly in response to distinct nitrogen and phosphate ratios. In turn, nutrient ratios may have been affected by the magnitude of fish feed input. Taken together, our results show that, even in strongly anthropogenically impacted aquatic ecosystems, the structure of bacterial communities is mainly driven by the natural seasonality, with more subtle effects of aquaculture-related factors.

Alkyltrimethylammonium (ATMA) surfactants as cyanocides - Effects on photosynthesis and growth of cyanobacteria.

Cyanobacteria and their toxins present potential hazard to consumers of water from lakes, reservoirs and rivers, thus their removal via water treatment or at the source, is essential. Here, we report that alkyltrimethylammonium (ATMA) surfactants, such as octadecyltrimethylammonium (ODTMA) bromide, act as cyanocides that efficiently inhibit photosynthesis and growth of cyanobacteria. Green algae were found less sensitive than cyanobacteria to ATMA compounds. Fluorescence measurements and microscopic observations demonstrated that cyanobacteria cells (Aphanizomenon or Microcystis) disintegrate and lose their metabolic activity (photosynthesis) upon exposure to ATMA bromides (estimated ED50(1hr) ranged between 1.5 and 7 µM for ODTMA-Br or hexadecyltrimethylammonium (HDTMA) bromide). Other ATMA compounds, such as tetradecyltrimethylammonium (TDTMA) or dodecyltrimethylammonium (DDTMA) bromides had similar inhibitory effect but their toxicity to cyanobacteria (measured as ED50(1hr) for photosynthetic efficiency) decreased, as the length of the alkyl chain decreased. All ATMA compounds used in this study showed lower toxicity to green algae than to cyanobacteria. A toxicity mechanism for ATMA cations is proposed, based on real time fluorescence signals and on alteration of cell ultra-structure revealed by electron microscopy. The present study sheds light on the toxic effect of ATMA surfactants on cyanobacteria and its potential application for controlling the occurrence of cyanobacterial bloom in lakes, reservoirs or rivers to secure the safety of drinking water and to mitigate and manage bloom events.

Recovery of Microcystis surface scum following a mixing event: Insights from a tank experiment.

Cyanobacteria of the genus Microcystis produces surface scum that negatively affects water quality in inland waters. This scum layer can be disintegrated and vertically dispersed by external forces (e.g., wind mixing), followed by reformation of surface scum as buoyant Microcystis colonies migrate upward. However, the recovery dynamics of Microcystis surface scum following a strong mixing event have rarely been studied. Here, we used a tank experiment to investigate the process of Microcystis surface scum recovery after a mixing event with focus on dynamics of colonies of different size classes and their contribution to that process. Microcystis colony size distribution and colony volume concentration was measured using a laser in-situ scattering and transmissometry instrument. The dynamics of Microcystis in the water column and upward colony migration velocity were strongly dependent on colony size. Larger colonies (>180 µm) with fast upward migration rates contributed the most to surface scum formation shortly after turbulence subsided. The contribution of slowly migrating smaller colonies to scum formation was observed over notably longer time. The estimated floating velocities of large colonies ranged 0.15 to 0.46 m h-1 depending on colony size and were 5-15 times higher than those of smaller colonies (~0.03 m h-1). The changes in colony size distribution of Microcystis in the water column reflect the dynamics of surface scum. Analysis of size distribution of Microcystis colonies can be used for better understanding and prediction of Microcystis surface scum development in water bodies.

Multiannual variations in Microcystis bloom episodes - Temperature drives shift in species composition.

Cyanobacteria are notorious for producing water blooms and for toxin formation. Toxic cyanobacterial blooms present an ever-increasing serious threat to both the quality of drinking water and recreational uses and severely disrupt aquatic ecosystems, worldwide. In many cases, such blooms are dominated by toxic Microcystis sp. that produce a family of structurally similar hepatotoxins, known as microcystins (MCs). Here we present a retrospective analysis of Microcystis seasonal blooms from Lake Kinneret (Sea of Galilee, Israel) indicating that the population is composed of at least 25 different genotypes and two different chemo-types, whose relative abundance changes over decades. Based on a long-term record of biotic and abiotic parameters and laboratory experiments we propose that minor increase in water temperature, but not in salinity, may affect Microcystis community structure by changing the relative abundance of species/strains from toxic to less or non-toxic species.

The global Microcystis interactome.

Bacteria play key roles in the function and diversity of aquatic systems, but aside from study of specific bloom systems, little is known about the diversity or biogeography of bacteria associated with harmful cyanobacterial blooms (cyanoHABs). CyanoHAB species are known to shape bacterial community composition and to rely on functions provided by the associated bacteria, leading to the hypothesized cyanoHAB interactome, a coevolved community of synergistic and interacting bacteria species, each necessary for the success of the others. Here, we surveyed the microbiome associated with Microcystis aeruginosa during blooms in 12 lakes spanning four continents as an initial test of the hypothesized Microcystis interactome. We predicted that microbiome composition and functional potential would be similar across blooms globally. Our results, as revealed by 16S rRNA sequence similarity, indicate that M. aeruginosa is cosmopolitan in lakes across a 280° longitudinal and 90° latitudinal gradient. The microbiome communities were represented by a wide range of operational taxonomic units and relative abundances. Highly abundant taxa were more related and shared across most sites and did not vary with geographic distance, thus, like Microcystis, revealing no evidence for dispersal limitation. High phylogenetic relatedness, both within and across lakes, indicates that microbiome bacteria with similar functional potential were associated with all blooms. While Microcystis and the microbiome bacteria shared many genes, whole-community metagenomic analysis revealed a suite of biochemical pathways that could be considered complementary. Our results demonstrate a high degree of similarity across global Microcystis blooms, thereby providing initial support for the hypothesized Microcystis interactome.

Particle-Associated Microbial Community in a Subtropical Lake During Thermal Mixing and Phytoplankton Succession.

Ecosystem dynamics in monomictic lakes are characterized by seasonal thermal mixing and stratification. These physical processes bring about seasonal variations in nutrients and organic matter fluxes, affecting the biogeochemical processes that occur in the water column. Physical and chemical dynamics are generally reflected in seasonal structural changes in the phytoplankton and bacterio-plankton community. In this study, we analyzed, using 16S rRNA amplicon sequencing, the structure of the bacterial community associated with large particles (>20 µm) in Lake Kinneret (Sea of Galilee, Israel), and its associations to phytoplankton populations. The study was carried out during late winter and early spring, a highly dynamic period in terms of thermal mixing, nutrient availability, and shifts in phytoplankton composition. Structural changes in the bacterioplankton population corresponded with limnological variations in the lake. In terms of the entire heterotrophic community, the structural patterns of particle-associated bacteria were mainly correlated with abiotic factors such as pH, ammonia, water temperature and nitrate. However, analysis of microbial taxon-specific correlations with phytoplankton species revealed a strong potential link between specific bacterial populations and the presence of different phytoplankton species, such as the cyanobacterium Microcystis, as well as the dinoflagellates Peridinium and Peridiniopsis. We found that Brevundimonas, a common freshwater genus, and Bdellovibrio, a well-known Gram-negative bacteria predator, were positively associated to Microcystis, suggesting a potentially important role of these three taxa in the microbial ecology of the lake. Our results show that the dynamics of environmental abiotic conditions, rather than specific phytoplankton assemblages, are the main factors positively correlated with changes in the community structure as a whole. Nevertheless, some specific bacteria may interact and be linked with specific phytoplankton, which may potentially control the dynamic patterns of the microbial community.

The response of Microcystis aeruginosa strain MGK to a single or two consecutive H2 O2 applications.

Various approaches have been proposed to control/eliminate toxic Microcystis sp. blooms including H2 O2 treatments. Earlier studies showed that pre-exposure of various algae to oxidative stress induced massive cell death when cultures were exposed to an additional H2 O2 treatment. We examined the vulnerability of exponential and stationary-phase Microcystis sp. strain MGK cultures to single and double H2 O2 applications. Stationary cultures show a much higher ability to decompose H2 O2 than younger cultures. Nevertheless, they are more sensitive to an additional H2 O2 dose given 1-6 h after the first one. Transcript analyses following H2 O2 application showed a fast rise in glutathione peroxidase abundance (227-fold within an hour) followed by a steep decline thereafter. Other genes potentially engaged in oxidative stress were far less affected. Metabolic-related genes were downregulated after H2 O2 treatments. Among those examined, the transcript level of prk (encoding phosphoribulose kinase) was the slowest to recover in agreement with the decline in photosynthetic rate revealed by fluorescence measurements. Our findings shed light on the response of Microcystis MGK to oxidative stress suggesting that two consecutive H2 O2 applications of low concentrations are far more effective in controlling Microcystis sp. population than a single dose of a higher concentration.

Secondary Metabolites of Aeromonas veronii Strain A134 Isolated from a Microcystis aeruginosa Bloom.

Aeromonas veronii strain A134 was isolated from Microcystis aeruginosa colonies collected from Lake Kinneret (Sea of Galilee), Israel. The Aeromonas culture media inhibited the growth of M. aeruginosa (strain MGK). The crude extract of a large-scale culture of A. veronii A134 was separated in a few chromatographic steps to yield three new secondary metabolites, 9-chlorolumichrome (1), veronimide (2) and veronipyrazine (3), along with a known lumichrome and several known diketopiperazines. The structures of the new compounds were established by analyses of the data from 1D and 2D NMR experiments and HRMS data of the compounds, as well as a single-crystal x-ray analysis of synthetic 1. The structure elucidation and proposed biogenesis of the new compounds are described below.

Increased algicidal activity of Aeromonas veronii in response to Microcystis aeruginosa: interspecies crosstalk and secondary metabolites synergism.

Toxic Microcystis spp. blooms constitute a serious threat to water quality worldwide. Aeromonas veronii was isolated from Microcystis sp. colonies collected in Lake Kinneret. Spent Aeromonas media inhibits the growth of Microcystis aeruginosa MGK isolated from Lake Kinneret. The inhibition was much stronger when Aeromonas growth medium contained spent media from MGK suggesting that Aeromonas recognized its presence and produced secondary metabolites that inhibit Microcystis growth. Fractionations of the crude extract and analyses of the active fractions identified several secondary metabolites including lumichrome in Aeromonas media. Application of lumichrome at concentrations as low as 4 nM severely inhibited Microcystis growth. Inactivation of aviH in the lumichrome biosynthetic pathway altered the lumichrome level in Aeromonas and the extent of MGK growth inhibition. Conversely, the initial lag in Aeromonas growth was significantly longer when provided with Microcystis spent media but Aeromonas was able to resume normal growth. The longer was pre-exposure to Microcystis spent media the shorter was the lag phase in Aeromonas growth indicating the presence of, and acclimation to, secondary MGK metabolite(s) the nature of which was not revealed. Our study may help to control toxic Microcystis blooms taking advantage of chemical languages used in the interspecies communication.

Integrating chytrid fungal parasites into plankton ecology: research gaps and needs.

Chytridiomycota, often referred to as chytrids, can be virulent parasites with the potential to inflict mass mortalities on hosts, causing e.g. changes in phytoplankton size distributions and succession, and the delay or suppression of bloom events. Molecular environmental surveys have revealed an unexpectedly large diversity of chytrids across a wide range of aquatic ecosystems worldwide. As a result, scientific interest towards fungal parasites of phytoplankton has been gaining momentum in the past few years. Yet, we still know little about the ecology of chytrids, their life cycles, phylogeny, host specificity and range. Information on the contribution of chytrids to trophic interactions, as well as co-evolutionary feedbacks of fungal parasitism on host populations is also limited. This paper synthesizes ideas stressing the multifaceted biological relevance of phytoplankton chytridiomycosis, resulting from discussions among an international team of chytrid researchers. It presents our view on the most pressing research needs for promoting the integration of chytrid fungi into aquatic ecology.

Removal of cyanobacteria and cyanotoxins from lake water by composites of bentonite with micelles of the cation octadecyltrimethyl ammonium (ODTMA).

Cyanobacteria and their toxins present potential hazard to consumers of water from lakes, reservoirs and rivers, thus their removal via water treatment is essential. The capacity of nano-composites of Octadecyltrimethyl-ammonium (ODTMA) complexed with clay to remove cyanobacterial and their toxins from laboratory cultures and from lake water, was evaluated. Column filters packed with micelles of ODTMA complexed with bentonite and granulated were shown to significantly reduce the number of cyanobacteria cells or filaments and their corresponding toxins from laboratory cultures. Fluorescence measurements demonstrated that cyanobacteria cells lost their metabolic activity (photosynthesis) upon exposure to the micelle (ODTMA)-bentonite complex, or ODTMA monomers. The complex efficiently removed cyanobacteria toxins with an exceptional high removal rate of microcystins. The effectiveness of the complex in elimination of cyanobacteria was further demonstrated with lake water containing cyanobacteria and other phytoplankton species. These results and model calculations suggest that filters packed with granulated composites can secure the safety of drinking water in case of a temporary bloom event of toxic cyanobacteria.

Characterization of a new chytrid species parasitic on the dinoflagellate, Peridinium gatunense.

Only a few chytrid fungi have been reported as parasites of dinoflagellates. Among these reports, chytrids are periodically observed growing on the dinoflagellate, Peridinium gatunense, in Lake Kinneret (Sea of Galilee), Israel. Because of the distinctive roles of parasitic chytrid fungi in decreasing phytoplankton populations and in transforming inedible algae into chytrid biomass which zooplankton grazers can eat, characterizing dinoflagellate parasites contributes to our understanding of the sustainability of this important water resource. An undescribed chytrid parasite of P. gatunense from Lake Kinneret has recently been brought into pure culture (KLL_TL-060613), facilitating exploration of its infection process. To evaluate the ability of this chytrid to affect host populations, we determined the effect of: (1) temperature and light (or dark) on prevalence of infection and (2) host growth phase and parasite:host ratio on percentage of infection. The greatest amplification in host infection occurred in cultures grown in the dark at 25 C. The percentage of host cells infected increased as the availability of host cells compared to parasite cells increased. These results demonstrate that environmental factors influence the chytrid's potential to affect Peridinium gatunense populations. Because this chytrid had not been described taxonomically, we characterized its thallus morphology, development, zoospore ultrastructure and phylogenetic relationships. Zoospore ultrastructure was compatible with the Group II type zoospore characteristic of the family Chytridiaceae in the Chytridiales. Consistent with this observation, phylogenetic analyses of nuc 28S rDNA D1-D3 domains (28S) placed the chytrid in a clade among described taxa in the Chytridiaceae. Because thallus morphology was distinct from these other taxa, as well as other described parasites of dinoflagellates, this chytrid is described as a new genus and species, Dinochytrium kinnereticum.