Non-toxic Strains Research Articles

Nutrient Regulation of Relative Dominance of Cylindrospermopsin-Producing and Non-cylindrospermopsin-Producing Raphidiopsis raciborskii.

Raphidiopsis raciborskii (previously Cylindrospermopsis raciborskii) can produce cylindrospermopsin (CYN) which is of great concern due to its considerable toxicity to human and animals. Its CYN-producing (toxic) and non-CYN-producing (non-toxic) strains co-exist commonly in natural water bodies, while how their relative dominance is regulated has not been addressed. In this study, we combined field investigation with laboratory experiments to assessed the relationship between toxic and non-toxic R. raciborskii abundances under different nutrient levels. The rpoC1- and cyrJ-based qPCR was applied for quantifying total and toxic R. raciborskii abundances, respectively. The field survey showed that toxic R. raciborskii was detected in 97 of 115 reservoirs where its proportion ranged from 0.3% to 39.7% within the R. raciborskii population. Both total and toxic R. raciborskii abundances increased significantly with trophic level of these reservoirs, consistent with our monoculture and co-culture experiments showing in an increase in R. raciborskii growth with increasing nitrogen (N) or phosphorus (P) concentrations. In the monoculture experiments, growth rates of non-toxic and toxic strains from Australia or China were not significantly different under the same culture conditions. On the other hand, in the co-culture experiments, the toxic strains displayed a significantly faster growth than non-toxic strains under nutrient-replete conditions, resulting in an obvious shift toward the dominance by toxic strains from day 3 to the end of the experiments, regardless of the strain originating from Australia or China. The reverse was found under N- or P-limited conditions. Our results indicated that the toxic strains of R. raciborskii have a competitive advantage relative to the non-toxic strains in a more eutrophic world. In parallel to an increase in dominance, both toxic strains grown in the mixed population significantly increased CYN production under nutrient-replete conditions as compared to nutrient-limited conditions, suggesting that CYN may be of significance for ecological advantage of toxic R. raciborskii. These results highlight the importance of nutrient availability in regulating abundances and strain dominance of two genotypes of R. raciborskii. Our findings demonstrated that elevated nutrients would favor the growth of CYN-producing R. raciborskii and CYN production, leading to more blooms with higher toxicity at global scale.

Genome Streamlining, Plasticity, and Metabolic Versatility Distinguish Co-occurring Toxic and Nontoxic Cyanobacterial Strains of Microcoleus.

ABSTRACTHarmful cyanobacterial bloom occurrences have increased worldwide due to climate change and eutrophication, causing nuisance and animal deaths. Species from the benthic cyanobacterial genus Microcoleus are ubiquitous and form thick mats in freshwater systems, such as rivers, that are sometimes toxic due to the production of potent neurotoxins (anatoxins). Anatoxin-producing (toxic) strains typically coexist with non-anatoxin-producing (nontoxic) strains in mats, although the reason for this is unclear. To determine the genetic mechanisms differentiating toxic and nontoxic Microcoleus, we sequenced and assembled genomes from 11 cultures and compared these to another 31 Microcoleus genomes. Average nucleotide identities (ANI) indicate that toxic and nontoxic strains are distinct species (ANI, <95%), and only 6% of genes are shared across all 42 genomes, suggesting a high level of genetic divergence among Microcoleus strains. Comparative genomics showed substantial genome streamlining in toxic strains and a potential dependency on external sources for thiamine and sucrose. Toxic and nontoxic strains are further differentiated by an additional set of putative nitrate transporter (nitrogen uptake) and cyanophycin (carbon and nitrogen storage) genes, respectively. These genes likely confer distinct competitive advantages based on nutrient availability and suggest nontoxic strains are more robust to nutrient fluctuations. Nontoxic strains also possess twice as many transposable elements, potentially facilitating greater genetic adaptation to environmental changes. Our results offer insights into the divergent evolution of Microcoleus strains and the potential for cooperative and competitive interactions that contribute to the co-occurrence of toxic and nontoxic species within mats.

Differential Labeling of Chemically Modified Peptides and Lipids among Cyanobacteria Planktothrix and Microcystis.

The cyanoHAB forming cyanobacteria Microcystis and Planktothrix frequently produce high intracellular amounts of microcystins (MCs) or anabaenopeptins (APs). In this study, chemically modified MCs and APs have been localized on a subcellular level in Microcystis and Planktothrix applying copper-catalyzed alkyne-azide cycloaddition (CuACC). For this purpose, three different non-natural amino acids carrying alkyne or azide moieties were fed to individual P. agardhii strains No371/1 and CYA126/8 as well as to M. aeruginosa strain Hofbauer showing promiscuous incorporation of various amino acid substrates during non-ribosomal peptide synthesis (NRPS). Moreover, CYA126/8 peptide knock-out mutants and non-toxic strain Synechocystis PCC6803 were processed under identical conditions. Simultaneous labeling of modified peptides with ALEXA405 and ALEXA488 and lipid staining with BODIPY 505/515 were performed to investigate the intracellular location of the modified peptides. Pearson correlation coefficients (PCC) obtained from confocal images were calculated between the different fluorophores and the natural autofluorescence (AF), and between labeled modified peptides and dyed lipids to investigate the spatial overlap between peptides and the photosynthetic complex, and between peptides and lipids. Overall, labeling of modified MCs (M. aeruginosa) and APs (P. agardhii) using both fluorophores revealed increased intensity in MC/AP producing strains. For Synechocystis lacking NRPS, no labeling using either ALEXA405 or ALEXA488 was observed. Lipid staining in M. aeruginosa and Synechocystis was intense while in Planktothrix it was more variable. When compared with AF, both modified peptides and lipids showed a heterologous distribution. In comparison, the correlation between stained lipids and labeled peptides was not increased suggesting a reduced spatial overlap.

Oral immunization of Escherichia albertii strain DM104 induces protective immunity against Shigella dysenteriae type 4 in mouse model.

The recent rise of antibiotic resistance and lack of an effective vaccine make the scenario of shigellosis alarming in developing countries like Bangladesh. In recent years, our group reported the vaccine efficacy of a non-pathogenic Escherichia albertii strain DM104 in different animal models, where an ocularly administered vaccine in the guinea pig eye model against Shigella dysenteriae type 4 challenge showed high protective efficacy and also induced a high titer of serum IgG against S. dysenteriae type 4 whole cell lysate (WCL) and LPS. In this study, we report further evaluation of the non-invasive and non-toxic environmental strain DM104 as a vaccine candidate against S. dysenteriae type 4 in mice model. Oral immunization of live DM104 bacterial strain demonstrated better protective immunity in mice model by showing 90% protection in mice against live S. dysenteriae type 4 lethal dose challenge and by inducing effective humoral and mucosal immune responses.

SxtA4+ and sxtA4- Genotypes Occur Together within Natural Pyrodinium bahamense Sub-Populations from the Western Atlantic.

Saxitoxin (STX) is a secondary metabolite and potent neurotoxin produced by several genera of harmful algal bloom (HAB) marine dinoflagellates. The basis for variability in STX production within natural bloom populations is undefined as both toxic and non-toxic strains (of the same species) have been isolated from the same geographic locations. Pyrodinium bahamense is a STX-producing bioluminescent dinoflagellate that blooms along the east coast of Florida as well as the bioluminescent bays in Puerto Rico (PR), though no toxicity reports exist for PR populations. The core genes in the dinoflagellate STX biosynthetic pathway have been identified, and the sxtA4 gene is essential for toxin production. Using sxtA4 as a molecular proxy for the genetic capacity of STX production, we examined sxtA4+ and sxtA4- genotype frequency at the single cell level in P. bahamense populations from different locations in the Indian River Lagoon (IRL), FL, and Mosquito Bay (MB), a bioluminescent bay in PR. Multiplex PCR was performed on individual cells with Pyrodinium-specific primers targeting the 18S rRNA gene and sxtA4. The results reveal that within discrete natural populations of P. bahamense, both sxtA4+ and sxtA4- genotypes occur, and the sxtA4+ genotype dominates. In the IRL, the frequency of the sxtA4+ genotype ranged from ca. 80–100%. In MB, sxtA4+ genotype frequency ranged from ca 40–66%. To assess the extent of sxtA4 variation within individual cells, sxtA4 amplicons from single cells representative of the different sampling sites were cloned and sequenced. Overall, two variants were consistently obtained, one of which is likely a pseudogene based on alignment with cDNA sequences. These are the first data demonstrating the existence of both genotypes in natural P. bahamense sub-populations, as well as sxtA4 presence in P. bahamense from PR. These results provide insights on underlying genetic factors influencing the potential for toxin variability among natural sub-populations of HAB species and highlight the need to study the genetic diversity within HAB sub-populations at a fine level in order to identify the molecular mechanisms driving HAB evolution.

The Influence of the Toxin-Producing Dinoflagellate, Alexandrium catenella (1119/27), on the Survival and Reproduction of the Marine Copepod, Acartia tonsa, During Prolonged Exposure

Copepods can feed on, and may regulate, the blooms of harmful algae (HA), and may also facilitate dinoflagellate blooms by inducing toxin production and through selective grazing. However, exposure to HA may also cause mortality and reproductive impairment in copepods, with detrimental effects at the population-scale. Here we present the toxin profile of the dinoflagellate, Alexandrium catenella (formerly Alexandrium tamarense), and examine how it affects the survival and reproduction of the cosmopolitan marine copepod, Acartia tonsa. Healthy adult copepods were exposed to mono-specific diets of toxic and non-toxic strains of A. catenella (1119/27 and 1119/19, respectively) and non-toxic Rhodomonas sp. for 10 days alongside unfed controls to examine how their survival was influenced by likely HA bloom conditions. Additional 2-day experiments examined how their egg production rate and hatching success were affected by food deprivation, toxic A. catenella, a non-toxic alternative and a mixture of toxic and non-toxic prey, at high and low concentrations. Survival of A. tonsa declined over the 10-day experiment in all treatments but was not significantly lower in the toxic A. catenella treatment; mortality was only significantly enhanced in the unfed animals, which showed 100% mortality after 9 days. Egg production rates and hatching success from females in the unfed and toxic A. catenella treatments were all significantly lower than values observed in females fed Rhodomonas sp. or non-toxic A. catenella. Animals offered 1,000 μg C L–1 of Rhodomonas sp. and a 50:50 mixture of toxic A. catenella and Rhodomonas sp. produced significantly more eggs than animals fed toxic A. catenella alone. These results were not apparent at prey concentrations of 100 μg C L–1. The percentages of eggs to successfully hatch from females offered mono-specific diets of toxic A. catenella were always close to zero. Collectively, our results indicate that adult female A. tonsa can acquire sufficient energy from toxic A. catenella to survive, but suffer reproductive impairment when feeding on this prey alone.

From the sxtA4 Gene to Saxitoxin Production: What Controls the Variability Among Alexandrium minutum and Alexandrium pacificum Strains?

Paralytic shellfish poisoning (PSP) is a human foodborne syndrome caused by the consumption of shellfish that accumulate paralytic shellfish toxins (PSTs, saxitoxin group). In PST-producing dinoflagellates such as Alexandrium spp., toxin synthesis is encoded in the nuclear genome via a gene cluster (sxt). Toxin production is supposedly associated with the presence of a 4th domain in the sxtA gene (sxtA4), one of the core genes of the PST gene cluster. It is postulated that gene expression in dinoflagellates is partially constitutive, with both transcriptional and post-transcriptional processes potentially co-occurring. Therefore, gene structure and expression mode are two important features to explore in order to fully understand toxin production processes in dinoflagellates. In this study, we determined the intracellular toxin contents of twenty European Alexandrium minutum and Alexandrium pacificum strains that we compared with their genome size and sxtA4 gene copy numbers. We observed a significant correlation between the sxtA4 gene copy number and toxin content, as well as a moderate positive correlation between the sxtA4 gene copy number and genome size. The 18 toxic strains had several sxtA4 gene copies (9–187), whereas only one copy was found in the two observed non-toxin producing strains. Exploration of allelic frequencies and expression of sxtA4 mRNA in 11 A. minutum strains showed both a differential expression and specific allelic forms in the non-toxic strains compared with the toxic ones. Also, the toxic strains exhibited a polymorphic sxtA4 mRNA sequence between strains and between gene copies within strains. Finally, our study supported the hypothesis of a genetic determinism of toxin synthesis (i.e., the existence of several genetic isoforms of the sxtA4 gene and their copy numbers), and was also consistent with the hypothesis that constitutive gene expression and moderation by transcriptional and post-transcriptional regulation mechanisms are the cause of the observed variability in the production of toxins by A. minutum.

Relative Effects of Varying Light Conditions on the Growth of Toxic and Non-Toxic Strains of &amp;lt;i&amp;gt;Microcystis aeruginosa&amp;lt;/i&amp;gt;

Harmful algae bloom constitutes a major problem facing water bodies particularly fresh and marine water system. Microcystis aeruginosa represents a major causative organism found in the water. Light plays a major role in the growth and variation of M. aeruginosa in water. What is still inadequate, is the data on the effects of different light conditions on the growth of Microcystis aeruginosa. In this study, two strains of Microcystis aeruginosa PCC 7806 (toxic strain) and PCC 7005 (non-toxic strain) were exposed to varying light intensities at 0, 30, 60, 120, 300, 350 μMol photon m-2·S-1 to measure their growth and pigmentation. This study observed similarities in growth and pigmentation between toxic strain (PCC 7806) and non-toxic (PCC 7005) strain. Also, at short exposure to light, Toxic strain was seen to be better tolerant of light than non-toxic strain, with non-toxic strain showing better ability to recover from light stress after twenty days of culture. These findings suggest that Toxic strain (PCC 7806) and Non-toxic strain (PCC 7005) of M. aeruginosa respond similarly with respect to growth but differ in their light retention capacity over time and this could be useful in predicting the possible conditions of algal bloom.

Comparison of biological methods to control Aphis fabae Scopoli (Hemiptera: Aphididae) on kalanchoe crops in East Africa

Aphids cause considerable damage to numerous crops all over the world and insecticides are the main means of controlling them, despite their detrimental impacts on human and environmental health. This study assessed the effectiveness of the parasitoid Aphidius colemani Viereck (Hymenoptera: Braconidae), a mixture of predatory ladybird beetles, Hippodamia variegata Goeze, Chilocorus calvus Chiccl, and Cheilomenes propinqua Mulsant (Coleoptera: Coccinellidae), and an entomopathogenic strain of Aspergillus flavus Link (Eurotiales: Trichocomaceae), collected locally in Tanzania, to control Aphis fabae Scopoli (Hemiptera: Aphididae). After assessing the predation and parasitism rates of these natural enemies at different aphid densities in laboratory experiments, their ability to control aphids on kalanchoe was assessed in a greenhouse experiment over two seasons. The largest number of A. fabae parasitized or consumed in the laboratory was found at a density of 160 aphids per predator, or parasitoid. At that density, an adult female of A. colemani parasitized 114 A. fabae per day, on average, and adults of C. calvus, H. variegata, and C. propinqua consumed 75, 72, and 85 aphids per day, respectively. A. flavus spores applied at 1 × 107 spores ml−1 reduced the aphid population by 7.9 and 12.6 times within 10 days in the first and second seasons of the greenhouse experiments, respectively, as opposed to 2.8 and 2.5 times by releasing a mixture of the ladybirds at a rate of 5 adults/m2, and by 3.3 and 9.5 times by releasing A. colemani at a rate of 2 adults/m2. This study confirmed the potential of these locally collected bio-control agents for controlling A. fabae. However, use of the isolated A. flavus strain was undermined by its production of aflatoxin. Further research is therefore required to tap into the potential of a non-toxic strain of A. flavus and/or other entomopathogenic fungi.

Invasion and toxin production by exotic nostocalean cyanobacteria (Cuspidothrix, Cylindrospermopsis, and Sphaerospermopsis) in the Nakdong River, Korea

The extent and frequency of harmful cyanobacterial blooms are increasing, owing to the climate change caused by global warming, and some harmful filamentous cyanobacteria that were first reported in the tropics are spreading to temperate regions, such as North America, Europe, and Northeast Asia. Although these exotic invasive cyanobacteria have a high toxigenic potential, they are not targeted in management plans in many countries. This study analyzed the occurrence of and potential toxin and off-flavor secondary metabolite production by invasive nostocalean cyanobacteria in the Nakdong River in Korea, which is a temperate region. The occurrence of four species belonging to three genera of cyanobacteria was confirmed in the Nakdong River. The quantities of cyanobacteria in the Nakdong River were mostly low, fewer than 1,000 cells mL−1. Twenty-four strains belonging to four species in three genera of cyanobacteria were isolated from the Nakdong River. Analysis revealed no off-flavor secondary metabolite production by any of the isolates, and those belonging to Cylindrospermopsis raciborskii, Sphaerospermopsis aphanizomenoides, and S. reniformis were identified as nontoxic strains. However, anatoxin-a production was observed in two of the eleven isolates of Cuspidothrix issatschenkoi. Given the sites and the timing of its occurrence, C. issatschenkoi had the highest potential for toxin production among the invasive nostocalean cyanobacteria appearing in the Nakdong River.

Methane production in harmful algal blooms collapsed water: The contribution of non-toxic Microcystis aeruginosa outweighs that of the toxic variety

Cyanobacterial blooms can promote CH4 production. The proportions of toxic and non-toxic Microcystis aeruginosa spatiotemporally vary in algal water. However, the comparative contributions of decomposed toxic and non-toxic M. aeruginosa to methane (CH4) production are poorly understood. Here, a 61-day microcosm study was conducted to simulate the collapse processes of toxic and non-toxic M. aeruginosa. The results illustrated that the decomposition of M. aeruginosa induced anaerobic and nutrient-rich conditions in the water. However, the presence of non-toxic M. aeruginosa promoted the release of protein-like substances and aromatic proteins that are readily utilized by methanogens. In comparison, the release of microcystins by toxic M. aeruginosa resulted in a distinctive microorganism community structure. The relative abundance of dominant methanogens in the sediment of the system containing toxic M. aeruginosa was markedly lower than that of the system containing the non-toxic strain at the end of incubation. Despite the fact that almost equal amounts of total carbon were present in the two systems, the CH4 production per unit particulate total carbon (identified as carbon content vs. algal biomass) in the system containing non-toxic M. aeruginosa was 1.74-fold higher than that in the system containing the toxic strain. Considering these observations, it is likely that the currently predicted CH4 mass in algal waters is inaccurate, and should be considered when estimating CH4 emissions in eutrophic freshwater.

Characteristics of Fluorescence Spectra, UV Spectra, and Specific Growth Rates during the Outbreak of Toxic Microcystis Aeruginosa FACHB-905 and Non-Toxic FACHB-469 under Different Nutrient Conditions in a Eutrophic Microcosmic Simulation Device

Microcystis aeruginosa is the dominant alga forming cyanobacteria blooms, the growth of which is limited by available nutrients. Thus, it is necessary to study cyanobacteria blooms and explore the growth of Microcystis aeruginosa under different nutrient conditions. In this paper, we take Microcystis aeruginosa, including toxic Freshwater Algae Culture of Hydrobiology Collection (FACHB)-905 and non-toxic FACHB-469 strains, into account. The strains were cultured using a simulation device under different nutrient conditions. Ultraviolet spectra, three-dimensional fluorescence spectra, and kinetic parameter indicators of the two species are studied. Compared to FACHB-469, the results show that the specific growth rate of FACHB-905 is much higher, in particular, FACHB-905 is the dominant species under low nutrient conditions. Furthermore, the UV spectral characteristics indicate that the molecular weight of dissolved organic matter in the culture tank of toxic FACHB-905 is greater than that of FACHB-469. Additionally, the humification index of toxic FACHB-905 is slightly higher as well, which suggests that it is more stable in the presence of dissolved organic matter during blooms. Therefore, the toxic Microcystis strain is more likely to become the dominant species in water blooms under lower eutrophic conditions and water blooms formed by the toxic Microcystis strain may be more difficult to recover from.

Molecular Mediation of Prion-like α-Synuclein Fibrillation from Toxic PFFs to Nontoxic Species.

Braak's theory described Parkinson's disease (PD) progression as prion-like α-synuclein (αSyn) spreading, which fundamentally subverts the understanding of pathogenesis. The pathological αSyn spreading pathway includes uptake, propagation, and release. However, the previous disease models were limitedly focusing on amyloid propagation/aggregation, which significantly impedes the mechanism exploration in spreading pathways and related therapeutic development. The spreading model can be achieved using recombinant αSyn preformed fibrils (PFFs), which seeds endogenous αSyn monomer to aggregation and causes substantial pathology and neurotoxicity. Here, we determined that dihydromyricetin (DHM), a natural flavonoid extracted from Ampelopsis grossedentata, can promote the fibrillization of prion-like PFF and induce propagation to form a distinct strain. Furthermore, administration of DHM significantly reduced prion-like PFF-induced propagation and neurotoxicity. The discovery of inducing infectious and neurotoxic PFF to a nontoxic strain resulting in neuron protection via promoting the fibrillization of PFF rather than inhibiting advances the understanding of the prion-like spreading mechanism and helps in developing treatments against PD and related α-synucleinopathies.

Toxic benthic freshwater cyanobacterial proliferations: Challenges and solutions for enhancing knowledge and improving monitoring and mitigation.

1. This review summarises knowledge on the ecology, toxin production, and impacts of toxic freshwater benthic cyanobacterial proliferations. It documents monitoring, management, and sampling strategies, and explores mitigation options. 2. Toxic proliferations of freshwater benthic cyanobacteria (taxa that grow attached to substrates) occur in streams, rivers, lakes, and thermal and meltwater ponds, and have been reported in 19 countries. Anatoxin- and microcystin-containing mats are most commonly reported (eight and 10 countries, respectively). 3. Studies exploring factors that promote toxic benthic cyanobacterial proliferations are limited to a few species and habitats. There is a hierarchy of importance in environmental and biological factors that regulate proliferations with variables such as flow (rivers), fine sediment deposition, nutrients, associated microbes, and grazing identified as key drivers. Regulating factors differ among colonisation, expansion, and dispersal phases. 4. New -omics-based approaches are providing novel insights into the physiological attributes of benthic cyanobacteria and the role of associated microorganisms in facilitating their proliferation. 5. Proliferations are commonly comprised of both toxic and non-toxic strains, and the relative proportion of these is the key factor contributing to the overall toxin content of each mat. 6. While these events are becoming more commonly reported globally, we currently lack standardised approaches to detect, monitor, and manage this emerging health issue. To solve these critical gaps, global collaborations are needed to facilitate the rapid transfer of knowledge and promote the development of standardised techniques that can be applied to diverse habitats and species, and ultimately lead to improved management.

Cyanobacterial Classification with the Toxicity Using MALDI Biotyper.

An abnormal growth of cyanobacteria in eutrophicated freshwaters can cause various environmental problems. In particular, Microcystis producing hepatotoxic cyclic heptapeptides microcystins (MCs) has been globally observed. Recent studies have demonstrated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) offers a rapid classification of cyanobacteria; however, they have not fully considered the toxicity yet. In this study, we have performed MALDI-TOF MS for intact cyanobacterial cells using Biotyper software and optimized their conditions to achieve cyanobacterial classification with the toxicity. The detection mass range used for Biotyper was extended to cover small molecules, but their intense ions were suppressed as a function of the used instrument Autoflex Speed, which enabled simultaneous observations of large molecular fingerprints and small MCs with comparable ion intensity. Hierarchical clustering of mass spectra obtained under the optimized conditions differentiated toxic and non-toxic clusters of Microcystis strains and furthermore formed a tight cluster of non-toxic strains possessing the MC biosynthesis gene mcyG. Spectral libraries were expanded to >30 genera (>80 strains) under the default and optimized conditions to improve the confidence of cyanobacterial classification. Consequently, spectral library searching allowed for characterization of cyanobacteria from a field sample as mixed toxic and non-toxic Microcystis cells, without isolating those cells.

Phylogenetic relationships and genetic divergence of paralytic shellfish toxin- and cylindrospermopsin- producing Cylindrospermopsis and raphidiopsis

Cylindrospermopsis and Raphidiopsis (C/R group) are closely related species responsible for cyanobacterial blooms worldwide. Paralytic shellfish toxins (PSTs) and cylindrospermopsins (CYNs) have been identified in different C/R group strains. However, the evolutionary relationship between PST- and CYN-producing strains has not been systematically evaluated. In this study, C/R group strains and their toxin biosynthesis genes were evaluated by phylogenetic analysis and sequence comparison. None of the tested strains are able to produce PSTs and CYNs simultaneously. The C/R group strains were clustered into five clades, including two non-toxic, two CYN-producing and one PST-producing clades. A high degree of similarity was observed for rpoC1 (> 96%) and ITS-L (> 97%) sequences within each clade with the exception of the ITS-L (87% to 100%) region in CYN-producing R. curvata, which has been shown to contain variable sequence insertions. Genomic analysis revealed that sxtY and sxtZ could be found in both toxic and non-toxic strains. The transposase gene IS4 was only observed in strains from the PST-producing clade. The sxt and cyr gene clusters share five gene families with similar functions. The amino acid sequences of the adenylyl-sulfate kinase genes, sxtO and cyrN, are more similar (45% to 81%) than other pairs of genes (8.0% to 40%). SxtO and CyrN proteins from C/R group strains forms an independent clade on the phylogenetic tree with a high degree of sequence similarity (78% to 100%). In conclusion, PST- and CYN- producing C/R group species can be classified into different clades based on their phylogenetic profile. The sxtO and cyrN genes have probably diverged from a single ancestral adenylyl-sulfate kinase gene, and may be specifically used for toxin biosynthesis in C/R group species.

Comparison of growth and toxicity responses between non-toxic and toxic strains of Prorocentrum hoffmannianum

We aimed to study the growth and toxicity responses of non-toxic (CCMP683) and toxic (CCMP2804) strains of Prorocentrum hoffmannianum under various nitrate and phosphate concentrations. The 2 strains were cultured in L1-Si medium with standard, depleted or 10-fold repleted nitrate or phosphate. CCMP683 cultured in standard L1-Si medium exhibited delayed growth. Nitrate or phosphate depletion decreased the cell density of both strains. Repletion of nitrate slightly increased the cell density of both strains. Repletion of phosphate also slightly increased the cell density of CCMP2804 but surprisingly decreased the cell density of CCMP683. Okadaic acid (OA) and its derivatives were not detected in CCMP683. OA was detected only in CCMP2804. Depletion of nitrate or phosphate increased the cellular concentration of OA, and repletion of nitrate or phosphate had no effect on the cellular concentration of OA. Correlation analysis indicated that the cellular concentration of OA was negatively correlated with cell density. Differences in the growth response to phosphate repletion and in the ability to produce OA suggest that the 2 strains may be good candidates for comparative studies related to phosphate metabolism and OA toxicity.

Role of potentially toxic cyanobacteria in crustacean zooplankton diet in a eutrophic lake

The coexistence of potentially toxic bloom-forming cyanobacteria (CY) and generally smaller-sized grazer communities has raised the question of zooplankton (ZP) ability to control harmful cyanobacterial blooms and highlighted the need for species-specific research on ZP-CY trophic interactions in naturally occurring communities. A combination of HPLC, molecular and stable isotope analyses was used to assess in situ the importance of CY as a food source for dominant crustacean ZP species and to quantify the grazing on potentially toxic strains of Microcystis during bloom formation in large eutrophic Lake Peipsi (Estonia). Aphanizomenon, Dolichospermum, Gloeotrichia and Microcystis dominated bloom-forming CY, while Microcystis was the major genus producing cyanotoxins all over the lake. Grazing studies showed that CY, and especially colonial CY, formed a significant, and also preferred component of algae ingested by the cladocerans Bosmina spp. and Daphnia spp. while this was not the case for the more selective calanoid copepod Eudiaptomus gracilis. Molecular analyses confirmed the presence of CY, including Microcystis, in ZP guts. Further analyses using qPCR targeting cyanobacterial genus-specific mcyE synthase genes indicated that potentially toxic strains of Microcystis can be ingested directly or indirectly by all the dominant crustacean grazers. However, stable isotope analyses indicated that little, if any, assimilation from ingested bloom-forming CY occurred. The study suggests that CY, and particularly Microcystis with both potentially toxic and non-toxic strains, can be widely ingested by cladoceran grazers during a bloom event with implications for control of CY abundance and for transfer of CY toxins through the food web.

Strain-specific responses of toxic and non-toxic Microcystis aeruginosa to exudates of heterotrophic bacteria

Although the cyanobacterium Microcystis is colonial during water blooms, isolated Microcystis strains become unicellular in culture. Studies indicate that heterotrophic bacteria can promote Microcystis colonies in culture, but little is known about the underlying mechanism or how widespread it is among bacteria. This study investigated the identity and colony-promoting effects of bacteria isolated from the 2014 Microcystis bloom in Lake Erie. Isolates were classified by their 16S rRNA gene sequences. Toxic and non-toxic cultures of M. aeruginosa were exposed to exudates of bacterial isolates, and their morphology, polysaccharide content, and reflectance spectra were compared to those of M. aeruginosa control cultures. Six isolates belonging to three genera enhanced the frequency or size of M. aeruginosa colonies in cultures where a dialysis barrier prevented direct contact between heterotrophic cells and M. aeruginosa cells. Toxic and non-toxic M. aeruginosa strains differed in how their morphology and optical properties responded to treatment. This study demonstrates that heterotrophic bacteria can promote colonial morphology in Microcystis without making physical contact with the Microcystis cells, as well as the first to indicate that toxic and non-toxic strains of the same morphospecies have different morphological and optical responses.

Predator-prey interactions between the ciliate Blepharisma americanum and toxic (Microcystis spp.) and non-toxic (Chlorella vulgaris, Microcystis sp.) photosynthetic microbes

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 83:211-224 (2019) - DOI: https://doi.org/10.3354/ame01913 Predator-prey interactions between the ciliate Blepharisma americanum and toxic (Microcystis spp.) and non-toxic (Chlorella vulgaris, Microcystis sp.) photosynthetic microbes Ian J. Chapman1,2, Daniel J. Franklin1, Andrew D. Turner3, Eddie J. A. McCarthy1, Genoveva F. Esteban1,* 1Bournemouth University, Department of Life and Environmental Sciences, Faculty of Science and Technology, Dorset, BH12 5BB, UK 2NSW Shellfish Program, NSW Food Authority, Taree, NSW 2430, Australia 3Centre for Environment, Fisheries and Aquaculture Science (CEFAS), Weymouth, Dorset, DT4 8UB, UK *Corresponding author: gesteban@bournemouth.ac.uk ABSTRACT: Despite free-living protozoa being a major factor in modifying aquatic autotrophic biomass, ciliate-cyanobacteria interactions and their functional ecological roles have been poorly described, especially with toxic cyanobacteria. Trophic relationships have been neglected and grazing experiments give contradictory evidence when toxic taxa such as Microcystis are involved. Here, 2 toxic Microcystis strains (containing microcystins), 1 non-toxic Microcystis strain and a non-toxic green alga, Chlorella vulgaris, were used to investigate predator-prey interactions with a phagotrophic ciliate, Blepharisma americanum. Flow cytometric analysis for microalgal measurements and a rapid ultra high performance liquid chromatography-tandem mass spectrometry protocol to quantify microcystins showed that non-toxic photosynthetic microbes were significantly grazed by B. americanum, which sustained ciliate populations. In contrast, despite constant ingestion of toxic Microcystis, rapid egestion of cells occurred. The lack of digestion resulted in no significant control of toxic cyanobacteria densities, a complete reduction in ciliate numbers, and no observable encystment or cannibalistic behaviour (gigantism). Individual B. americanum morphological responses (biovolume and cell width) showed a significant decrease over time when sustained on non-toxic Microcystis compared to grazed C. vulgaris populations, supporting previous studies that cyanobacteria may be a relatively poor source of nutrition. Results here provide insight into the ecological interactions of ciliates and cyanobacteria, and for the first time B. americanum is shown to have the capacity to suppress potentially bloom-forming cyanobacteria. However, grazing can be significantly altered by the presence of microcystins, which could have an impact on bloom dynamics and overall community structure. KEY WORDS: Ciliate-cyanobacteria interactions · Toxic cyanobacteria · Ciliate · Grazing · Microcystis · Microcystins Full text in pdf format PreviousNextCite this article as: Chapman IJ, Franklin DJ, Turner AD, McCarthy EJA, Esteban GF (2019) Predator-prey interactions between the ciliate Blepharisma americanum and toxic (Microcystis spp.) and non-toxic (Chlorella vulgaris, Microcystis sp.) photosynthetic microbes. Aquat Microb Ecol 83:211-224. https://doi.org/10.3354/ame01913 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 83, No. 3. Online publication date: September 19, 2019 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2019 Inter-Research.