Combined methods elucidate the multi-organ toxicity of cylindrospermopsin (CYN) on Daphnia magna.

Global water bodies are now at risk from inevitable cyanobacterial blooms and their production of multiple cyanotoxins, in particular cylindrospermopsin (CYN). However, research on the CYN toxicity and its molecular mechanisms is still limited, whilst the responses of aquatic species against CYN are uncovered. By integrating behavioral observations, chemical detections and transcriptome analysis, this study demonstrated that CYN exerted multi-organ toxicity to model species, Daphnia magna. The present study confirmed that CYN could cause protein inhibition by undermining total protein contents, and altered the gene expression related to proteolysis. Meantime, CYN induced oxidative stress by increasing reactive oxygen species (ROS) level, decreasing the glutathione (GSH) concentration, and interfered with protoheme formation process molecularly. Neurotoxicity led by CYN was solidly determined by abnormal swimming patterns, reduced acetylcholinesterase (AChE), and downward expression of muscarinic acetylcholine receptor (CHRM). Importantly, for the first time, this research determined CYN directly interfered with energy metabolism in cladocerans. CYN distinctively reduced filtration and ingestion rate by targeting on heart and thoracic limbs, which declined the energy intake, and could be further displayed by the reduction of motional strength and the trypsin concentration. These phenotypic alterations were supported by transcriptomic profile, including the down-regulation of oxidative phosphorylation and ATP synthesis. Moreover, CYN was speculated to trigger the self-defense responses of D. magna, known as "abandon-ship" by moderating lipid metabolism and distribution. This study, overall, comprehensively demonstrated the CYN toxicity and the responses of D. magna against it, which is of great significance to the advancements of CYN toxicity knowledge.

Chlorophyll f production in two new subaerial cyanobacteria of the family Oculatellaceae.

Chlorophyll (Chl) f was recently identified in a few cyanobacteria as the fifth chlorophyll of oxygenic organisms. In this study, two Leptolyngbya-like strains of CCNU0012 and CCNU0013 were isolated from a dry ditch in Chongqing city and a brick wall in Mount Emei Scenic Area in China, respectively. These two strains were described as new species: Elainella chongqingensis sp. nov. (Oculatellaceae, Synechococcales) and Pegethrix sichuanica sp. nov. (Oculatellaceae, Synechococcales) by the polyphasic approach based on morphological features, phylogenetic analysis of 16S rRNA gene and secondary structure comparison of 16S-23S internal transcribed spacer domains. Both strains produced Chl a under white light (WL) but additionally induced Chl f synthesis under far-red light (FRL). Unexpectedly, the content of Chl f in P. sichuanica was nearly half that in most Chl f-producing cyanobacteria. Red-shifted phycobiliproteins were also induced in both strains under FRL conditions. Subsequently, additional absorption peak beyond 700 nm in the FRL spectral region appeared in these two strains. This is the first report of Chl f production induced by FRL in the family Oculatellaceae. This study not only extended the diversity of Chl f-producing cyanobacteria but also provided precious samples to elucidate the essential binding sites of Chl f within cyanobacterial photosystems.

Genome Evolution of Filamentous Cyanobacterium Nostoc Species: From Facultative Symbiosis to Free Living.

In contrast to obligate bacteria, facultative symbiotic bacteria are mainly characterized by genome enlargement. However, the underlying relationship of this feature with adaptations to various habitats remains unclear. In this study, we used the global genome data of Nostoc strains, including 10 novel genomes sequenced in this study and 26 genomes available from public databases, and analyzed their evolutionary history. The evolutionary boundary of the real clade of Nostoc species was identified and was found to be consistent with the results of polyphasic taxonomy. The initial ancestral species of Nostoc was demonstrated to be consistent with a facultative symbiotic population. Further analyses revealed that Nostoc strains tended to shift from facultative symbiosis to a free-living one, along with an increase in genome sizes during the dispersal of each exterior branch. Intracellular symbiosis was proved to be essentially related to Nostoc evolution, and the adaptation of its members to free-living environments was coupled with a large preference for gene acquisition involved in gene repair and recombination. These findings provided unique evidence of genomic mechanisms by which homologous microbes adapt to distinct life manners and external environments.

Phylogenetic insights into chroococcus-like taxa (Chroococcales, Cyanobacteria), describing Cryptochroococcus tibeticus gen. nov. sp. nov. and Limnococcus fonticola sp. nov. from Qinghai-Tibet plateau.

Several coccoid cyanobacterial strains, morphologically similar to typical characteristics of Chroococcus, from the Qinghai-Tibet Plateau were isolated and characterized using a polyphasic approach including morphological and molecular information. Morphological characteristics, the phylogeny based on 16S rRNA gene, and 16S-23S internal transcribed spacer secondary structures support establishing a novel Chroococcus-like genus, Cryptochroococcus gen. nov., as well as Limnococcus fonticola sp. nov. Limnococcus is phylogenetically included in Chroococcaceae and has irregularly arranged thylakoids. Therefore, it should no longer be a member of Merismopediaceae (Synechococcales). The phylogeny based on the 16S rRNA gene revealed that Chroococcus-associated genera were monophyletic.

Four decades of progress in cylindrospermopsin research: The ins and outs of a potent cyanotoxin.

The cyanotoxin cylindrospermopsin (CYN), a toxic metabolite from cyanobacteria, is of particular concern due to its cosmopolitan occurrence, aquatic bioaccumulation, and multi-organ toxicity. CYN is the second most often recorded cyanotoxin worldwide, and cases of human morbidity and animal mortality are associated with ingestion of CYN contaminated water. The toxin poses a great challenge for drinking water treatment plants and public health authorities. CYN, with the major toxicity manifested in the liver, is cytotoxic, genotoxic, immunotoxic, neurotoxic and may be carcinogenic. Adverse effects are also reported for endocrine and developmental processes. We present a comprehensive review of CYN over the past four decades since its first reported poisoning event, highlighting its global occurrence, biosynthesis, toxicology, removal, and monitoring. In addition, current data gaps are identified, and future directions for CYN research are outlined. This review is beneficial for understanding the ins and outs of this environmental pollutant, and for robustly assessing health hazards posed by CYN exposure to humans and other organisms.

Geosmin production and polyphasic characterization of Oscillatoria limosa Agardh ex Gomont isolated from the open canal of a large drinking water system in Tianjin City, China.

Taste and odor (T & O) episodes always cause strong effects on drinking water supply system. Luanhe River diversion into Tianjin City in China is an important drinking water resource. Massive growth of a benthic filamentous cyanobacterium with geosmin production in the open canal caused a strong earthy odor episode in Tianjin. On the basis of the morphological and molecular identification of this cyanobacterium as Oscillatoria limosa Agardh ex Gomont, the genetic basis for geosmin biosynthesis and factors influencing growth and geosmin production of O. limosa CHAB 7000 were studied in this work. A 2268-bp open reading frame, encoding 755 amino acids, was amplified and characterized as the geosmin synthase gene (geo), followed by a cyclic nucleotide-binding protein gene (cnb). Phylogenetic analysis implied that the evolution of the geosmin genes in O. limosa CHAB 7000 might involve a horizontal gene transfer event. Examination on the growth and geosmin production of O. limosa CHAB 7000 at different light intensities showed that the maximum geosmin production was observed at 10µmol photons m-2s-1, while the optimum growth was at 60µmol photons m-2s-1. Under three temperature conditions (15°C, 25°C, and 35°C), the maximum growth and geosmin production were observed at 25°C. Most amounts of geosmin were retained in cells during the growth phase, but high temperature and low light intensity increased the release of geosmin into the medium, implying that O. limosa CHAB 7000 had a high potential harm for the release of geosmin from its cells at these adverse conditions.

Dynamics and polyphasic characterization of odor-producing cyanobacterium Tychonema bourrellyi from Lake Erhai, China.

The previous studies indicated that Tychonema-like strains from Lake Erhai could release geosmin so that the species was listed as the potential harmful cyanobacteria influencing the drinking water safety around Lake Erhai. But, the dynamics and biological information of this species were too limited. In this study, the polyphasic approach was used to reveal its biological characterization and the dynamics in Lake Erhai. The characters of trichomes, including filaments with solitary or bundle state, reddish-brown or blue-green color, planktonic habitat, and presence of keritomized content, were examined by the microscopic method. The 16S rDNA sequences of these strains were used for phylogenetic analysis and molecular identification. The strains were morphologically classified as Tychonema bourrellyi, and geosmin and ß-ionone were identified as the major volatile substances using gas chromatography-mass spectrometry (GC-MS) analysis. No strains of T. bourrellyi were found to produce microcystin by the HPLC and mcy gene approaches. Cell numbers at 12 sampling sites in Lake Erhai were shown as an average of 3 × 10(4) cells L(-1) in 2009 and 2010. The obvious peaks occurred in July and August each year. This was the first report on occurrence of T. bourrellyi from outside of Europe, and T. bourrellyi was also a newly recorded species in China. Such a result demonstrated that T. bourrellyi could distribute extending from cold waters in North Europe to the warm waters in subtropical regions. It was interesting to observe the coincidence of the occurrence of T. bourrellyi with slightly eutrophicated waters since Lake Erhai had been regarded as an early phase of eutrophicated lake.

Elucidating the toxicity targets of ß-ionone on photosynthetic system of Microcystis aeruginosa NIES-843 (Cyanobacteria).

In order to explore the potential targets of toxicity of ß-ionone on the photosynthetic system of Microcystis aeruginosa, the polyphasic rise in chlorophyll a (Chl a) fluorescence transient and transcript expression for key genes in photosystem II (PSII) of M. aeruginosa NIES-843 were studied. The EC50 value of ß-ionone on M. aeruginosa NIES-843 was found to be 21.23±1.87 mg/L. It was shown that ß-Ionone stress can lead to a decrease in pigment content of M. aeruginosa NIES-843 cells, and that carotenoids were more sensitive to ß-ionone stress than Chl a. The normalized Chl a fluorescence transients were slightly decreased at 6.67 and 10 mg/L ß-ionone, but significantly increased at 15, 22.5 and 33.75 mg/L. There was no significant variation on transcript expression of psbA and psbO at a concentration of 6.67 mg/L ß-ionone, but they were down-regulated at 22.5 mg/L. Ultrastructural examination by transmission electron microscopy indicated that the thylakoids were distorted, and the thylakoid membrane stacks began to collapse when M. aeruginosa NIES-843 was exposed to ß-ionone at a concentration of 22.5 and 33.75 mg/L. Our results indicate that the reaction centre of PS II and the electron transport at the acceptor side of PS II are the targets responsible for the toxicity of ß-ionone on the PS II of M. aeruginosa NIES-843.

Responses of Synechocystis sp. PCC 6803 (cyanobacterium) photosystem II to pyrene stress.

In order to explore the mechanism of acute toxicity for pyrene to cyanobacterial organisms, the responses of Synechocystis sp. PCC 6803 photosystem II (PS II) under pyrene stress were studied. The results showed there was no significant difference about the oxygen evolution under 0.125 mg/L pyrene stress when compared with control, but it was significantly lower than control at 0.625 mg/L pyrene. Polyphasic chlorophyll-a fluorescence transients in cells of Synechocystis sp. PCC 6803 exhibited a typical increase including O, J, I, and P phases. Fluorescence yield at phases J, I and P declined slightly at 0.125 and 0.625 mg/L pyrene, and significantly lower than control at 3.125 mg/L. According to the parameters deviated from JIP-test, no modification was induced by pyrene both at the donor side and at the acceptor side of PS II, and the reaction centre of PS II is the primary damaging target. Based on the expressing of four key genes (psbA, psbB, psbC and psbO) of PS II, only psbA showed significant difference at 3.125 mg/L pyrene when compared with control.

Towards clarification of the inhibitory mechanism of wheat bran leachate on Microcystis aeruginosa NIES-843 (Cyanobacteria): physiological responses.

Wheat bran leachate (WBL) has been shown to have an inhibitory effect on Microcystis aeruginosa in this study. In order to explore the inhibitory mechanism of WBL on M. aeruginosa, physiological responses of M. aeruginosa NIES-843 under the WBL stress were studied. The expressions of six important genes related to the D1 protein of photosynthetic processes (psbA), synthesis of microcystins (mcyB), antioxidant protein peroxiredoxin (prx), synthesis of fatty acid (fabZ) and the repair of biological macromolecules (recA, grpE) were studied. Under the WBL stress, no significant change was found in expressions of both grpE and recA, but expressions of psbA, fabZ and prx were shown to be down-regulated, and slight up-regulated expression was found in mcyB. It was shown that oxygen evolution of M. aeruginosa NIES-843 was significantly depressed, and intracellular ATP contents became lower, after exposure to WBL. Similarly, maximum electron transport rates of photosynthetic activities decreased significantly, but intracellular reactive oxygen species levels boosted dramatically under the WBL stress, and cell lysis was observed. Therefore, it is suggested that photosynthetic systems and membranes were the potential targets of toxicity of WBL on M. aeruginosa, and the oxidative damage is an important mechanism explaining the inhibitory effect of WBL on M. aeruginosa.