A laboratory based exposure of Microcystis and Oscillatoria cyanobacterial isolates to heterotrophic bacteria.

Biological control of cyanobacteria is a viable means of controlling nuisance bloom occurrences; however the majority of studies done are against Microcystis sp., with a commonly lytic effect caused. Filamentous cyanobacteria such as Oscillatoria are not as extensively studied in this area of biological control and are often part of Microcystis dominated blooms. This study employed heterotrophic bacterial isolates selected from bloom waters that indicated potential predatory behaviour against both filamentous and colonial cyanobacterial isolates. In comparison to a known Bacillus isolate, which is often reported among bacterial control agents, three other bacteria isolates were tested as control agents against non-axenic Oscillatoria and Microcystis cyanobacterial cultures. Assessments of cyanobacterial cell responses to the bacteria were conducted through water chemistry, chlorophyll a, alkaline phosphatase activity, microscopy and cyanotoxin measurements. The changes in these parameters were compared to untreated cyanobacterial cultures where no bacteria were added. The study found that at ratios of bacteria half that of Microcystis, minimal changes in chlorophyll a were observed, whilst Oscillatoria showed a decreased chlorophyll a more in the presence of isolates 1 and 3w. The assessment of alkaline phosphatase activity showed decreased activity in both cyanobacterial isolates exposed to the bacteria, relative to the untreated control sample. Microscopy analysis through fluorescence indicated that the attachment of the bacteria to the surface of the cyanobacteria hampered with the fluorescence and scanning electron microscopy indicated that the cells were damaged by the addition of the bacterial isolates. Cyanotoxin detection through the ELISA kit testing indicated that there was toxin reduction in samples treated with the bacterial isolates, with the highest reduction being close to 60% in the case of Microcystis sp. treated with isolate 3w. Similar reductions were noted in the filamentous cyanobacterium Oscillatoria, in the presence of isolate 1.

Characterization of phototrophic microorganisms and description of new cyanobacteria isolated from the saline-alkaline crater-lake Dziani Dzaha (Mayotte, Indian Ocean).

The saline-alkaline crater-lake Dziani Dzaha (Mayotte, Indian Ocean) is dominated by the bloom-forming cyanobacterium Arthrospira. However, the rest of the phototrophic community remains underexplored because of their minute dimension or lower biomass. To characterize the phototrophic microorganisms living in this ecosystem considered as a modern analog of Precambrian environments, several strains were isolated from the water column and stromatolites and analyzed using the polyphasic approach. Based on morphological, ultrastructural and molecular (16S rRNA gene, 18S rRNA gene, 16S-23S internal transcribed spacer (ITS) region and cpcBA-IGS locus) methods, seven filamentous cyanobacteria and the prasinophyte Picocystis salinarum were identified. Two new genera and four new cyanobacteria species belonging to the orders Oscillatoriales (Desertifilum dzianense sp. nov.) and Synechococcales (Sodalinema komarekii gen. nov., sp. nov., Sodaleptolyngbya stromatolitii gen. nov., sp. nov. and Haloleptolyngbya elongata sp. nov.) were described. This approach also allowed to identify Arthrospira fusiformis with exclusively straight trichomes instead of the spirally coiled form commonly observed in the genus. This study evidenced the importance of using the polyphasic approach to solve the complex taxonomy of cyanobacteria and to study algal assemblages from unexplored ecosystems.

Widespread anatoxin-a detection in benthic cyanobacterial mats throughout a river network.

Benthic algae fuel summer food webs in many sunlit rivers, and are hotspots for primary and secondary production and biogeochemical cycling. Concerningly, riverine benthic algal assemblages can become dominated by toxic cyanobacteria, threatening water quality and public health. In the Eel River in Northern California, over a dozen dog deaths have been attributed to cyanotoxin poisonings since 2000. During the summers of 2013-2015, we documented spatial and temporal patterns of cyanotoxin concentrations in the watershed, showing widespread distribution of anatoxin-a in benthic cyanobacterial mats. Solid phase adsorption toxin tracking (SPATT) samplers were deployed weekly to record dissolved microcystin and anatoxin-a levels at 10 sites throughout the watershed, and 187 Anabaena-dominated or Phormidium-dominated cyanobacterial mat samples were collected from 27 locations to measure intracellular anatoxin-a (ATX) and microcystins (MCY). Anatoxin-a levels were higher than microcystin for both SPATT (mean MCY = 0.8 and ATX = 4.8 ng g resin-1 day-1) and cyanobacterial mat samples (mean MCY = 0.074 and ATX = 1.89 µg g-1 DW). Of the benthic mats sampled, 58.9% had detectable anatoxin-a (max = 70.93 µg g-1 DW), while 37.6% had detectable microcystins (max = 2.29 µg g-1 DW). SPATT cyanotoxin levels peaked in mid-summer in warm mainstem reaches of the watershed. This is one of the first documentations of widespread anatoxin-a occurrence in benthic cyanobacterial mats in a North American watershed.

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.

Recruitment of cyanobacteria from the sediments in the eutrophic Shanzi Reservoir.

This study investigated the impact of four environmental factors on the recruitment of cyanobacteria from bottom sediments in the eutrophic Shanzi Reservoir. Temperature and light were identified as the key determinants for the recruitment of Microcystis and Oscillatoria. Cyanobacteria became dominant at higher temperature (20 °C) and light intensity (2000 lx) and Microcystis and Oscillatoria were the major species. Detailed recruitment simulation undertaken with the respective gradients of temperature and light suggested that both Microcystis and Oscillatoria are temperature sensitive and that their critical temperature point was 10 °C. However, distinct light impacts were observed only on Microcystis. The recruitment of Oscillatoria was light independent, whereas Microcystis had a positive relationship with light intensity. Physical disturbance promoted Microcystis recruitment and also affected the structure of the recruited cyanobacterial community at the water-sediment interface, based on quantitative polymerase chain reaction (qPCR) and phylogenetic analysis.

[Simultaneous removal of algae and its odorous metabolite dimethyl trisulfide in water by potassium ferrate].

Co-removal of oscillatoria algae and its potential odorous metabolite dimethyl trisulfide (DMTS) in simulated algae-laden alkaline source water by potassium ferrate (K2FeO4) was investigated in contrast to potassium permanganate (KMnO4) pre-oxidation followed by polyferric chloride (PFC) under varying conditions, including pH, initial oxidant dosage and turbidity. Based on the pre-comparison with PFC, the optimal dosage of PFC in the combined KMnO4 pre-oxidation-PFC treatment was determined. Potassium ferrate resulted in 92.4% removal of algae, higher than PFC when the dosage was equivalent as measured by Fe and KMnO4 showed obviously positive effect as a coagulation aid. Degradation of dimethyl trisufide (92.5%) by potassium ferrate was better than the pre-oxidation of potassium permanganate (74.6%), and the treatment time was decreased from 10 min to 1 min.

Molecular identification and ultrastructural and phylogenetic studies of cyanobacteria from association with the white sea hydroid Dynamena pumila (L., 1758).

Three new cyanobacterial strains, that have been previously purified from the hydroid Dynamena pumila (L., 1758), isolated from the White Sea, were studied using scanning and transmission electron microscopy methods and were characterized by using almost complete sequence of the 16S rRNA gene, internal transcribed spacer 16S-23S rRNA, and part of the gene for 23S rRNA. The full nucleotide sequences of the rRNA gene clusters were deposited to GenBank (HM064496.1, GU265558.1, JQ259187.1). Comparison of rRNA gene cluster sequences of Synechococcus cyanobacterium 1Dp66E-1, Oscillatoriales cyanobacterium 2Dp86E, and Nostoc sp. 10Dp66E with all sequences present at the GenBank shows that these cyanobacterial strains do not have 100% identity with any organisms investigated previously. Furthermore, for the first time heterotrophic bacterium, associated with Nostoc sp. 10Dp66E, was identified as a member of the new phylum Gemmatimonadetes, genus of Gemmatimonas (GenBank accession number is JX437625.1). Phylogenetic analysis showed that cyanobacterium Synechococcus sp. 1Dp66E-1 forms the unique branch and belongs to a cluster of Synechococcus, including freshwater and sea strains. Oscillatoriales cyanobacterium 2Dp86E belongs to a cluster of Leptolyngbya strains. Isolate Nostoc sp. 10Dp66E forms unique branch and belongs to a cluster of the genus Nostoc, with the closest relative of Nostoc commune isolates.

Removal of Pb²âº from aqueous system by live Oscillatoria laete-virens (Crouan and Crouan) Gomont isolated from industrial effluents.

Cyanobacteria have been found to be potential biosorbents of metal ions from waste water. The Pb²âº removal capacity of growing cells of indigenous cyanobacterium Oscillatoria laete-virens (Crouan and Crouan) Gomont was studied under batch experiments and it was found capable of removing Pb²âº of lower concentrations (below 100 mg L⁻¹). The effects of different concentrations of Pb²âº, on the growth rate of alga were also evaluated. The research parameters include the pH of the solution, contact time, initial concentration of Pb²âº, and culture density. Of the parameters studied, the pH of the solution was found to be the most crucial. The removal of Pb²âº peaked at an initial pH of 5. The data obtained from the equilibrium experiments were found well fitting with the Langmuir isotherm with a maximum sorptive capacity (q(max)) of 20.36 mg g⁻¹, indicating a good biosorbtive potential of growing cells. This was confirmed using scanning electron microscope and energy dispersive X-ray analysis, which showed the adsorption of lead on the surface of the cell. The species could tolerate a concentration as high as 60 mg L⁻¹ of Pb²âº. It was observed that the removal obeyed the pseudo-second-order kinetics. The percentage removal was found to decrease with increasing metal concentration, from 10 to 100 mg L⁻¹. FTIR analysis indicates the involvement of amino, carboxylic and amide groups in the sorption process. Among the desorbing agents evaluated, an efficient recovery of 90.2 % was achieved by HCl, in 24 h. Thus Oscillatoria laete-virens (Crouan and Crouan) Gomont seems to be a promising metal biosorbent for the treatment of Pb²âº, in waste waters.

Rapid determination of cytokinins and auxin in cyanobacteria.

Five cyanobacterial strains, Anabaena sp. Ck1, Oscillatoria sp. Ck2, Phormidium sp. Ck3, Chroococcidiopsis sp. Ck4, and Synechosystis sp. Ck5 were selected for their positive cytokinins-like activity using cucumber cotyledon bioassay and GUS assay in Arabidopsis ARR5::GUS. Classical cucumber cotyledon bioassay was modified for direct screening of cyanobacteria avoiding need for extraction and purification. Cytokinins from cyanobacteria were absorbed onto filter paper which was then assayed for cytokinins-like activity. A rapid chromatographic method was developed for the simultaneous determination of cytokinins and indole-3-acetic acid (IAA). Cyanobacterial biomass (50-100 mg) and cell-free culture filtrate were extracted in Bieleski buffer and purified by solid-phase extraction. The extract was used to determine phytohormones by ultra performance liquid chromatography and electrospray ionization-tandem mass spectrometry in positive and negative modes, respectively, with multiple reactions monitoring. Stable isotope-labeled cytokinins and IAA standards were added in the samples to follow recovery of the compounds and method validation. Five cytokinins determined in the selected strains were Zeatin (cis and trans isomers), Zeatin riboside, Dihydrozeatin riboside, and zeatin-o-glucoside. The strains were shown to accumulate as well as release the phytohormones.

Extracellular phycoerythrin-like protein released by freshwater cyanobacteria Oscillatoria and Scytonema sp.

During growth of the freshwater cyanobacteria, Oscillatoria sp. BTCC/A0004, and Scytonema sp. TISTR 8208, a pink pigment is released into the growth medium. The pigment from each source had a molecular weight of approximately 250 kDa and had adsorption maxima at 560 and 620 nm. These results suggest that pink pigment is a phycoerythrin-like protein. It inhibited the growth of green algae, Chlorella fusca and Chlamydomonas reinhardtii, but not other cyanobacteria or true bacteria. The concentration at which growth inhibition 50% occurred was 0.5, 6 and more than 10 mg ml(-1), respectively.

Adaptation of cyanobacteria to the sulfide-rich microenvironment of black band disease of coral.

Black band disease (BBD) is a cyanobacteria-dominated microbial mat that migrates across living coral colonies lysing coral tissue and leaving behind exposed coral skeleton. The mat is sulfide-rich due to the presence of sulfate-reducing bacteria, integral members of the BBD microbial community, and the sulfide they produce is lethal to corals. The effect of sulfide, normally toxic to cyanobacteria, on the photosynthetic capabilities of five BBD cyanobacterial isolates of the genera Geitlerinema (3), Leptolyngbya (1), and Oscillatoria (1) and six non-BBD cyanobacteria of the genera Leptolyngbya (3), Pseudanabaena (2), and Phormidium (1) was examined. Photosynthetic experiments were performed by measuring the photoincorporation of [(14)C] NaHCO(3) under the following conditions: (1) aerobic (no sulfide), (2) anaerobic with 0.5 mM sulfide, and (3) anaerobic with 0.5 mM sulfide and 10 microM 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU). All five BBD cyanobacterial isolates tolerated sulfide by conducting sulfide-resistant oxygenic photosynthesis. Five of the non-BBD cyanobacterial isolates did not tolerate sulfide, although one Pseudanabaena isolate continued to photosynthesize in the presence of sulfide at a considerably reduced rate. None of the isolates conducted anoxygenic photosynthesis with sulfide as an electron donor. This is the first report on the physiology of a culture of Oscillatoria sp. found globally in BBD.

[A laboratory model of the cyanobacterial mat from the Kotel'nikovskii hot spring (Baikal Region)].

A laboratory model of the cyanobacterial community of the Kotel'nikovskii hot spring (Baikal Region) was developed. A step-by-step description of the algocenosis formation along both the time and temperature gradient was given. The natural and laboratory mats were compared, and the major differences in the qualitative and quantitative composition of the cyanobacterial community were revealed. The laboratory algocenosis was stratified by species composition and was characterized by rapid replacement of the dominant cyanobacterial species depending on the temperature gradient. The formation of the community structure occurred over the 18 days of the experiment. In space and time, the sequence of species emergence in the cyanobacterial mat was as follows: Mastigocladus laminosus --> Phormidium tenue --> Ph. ambiguum --> Ph. valderiae. The species composition of the laboratory mat was similar to that of the natural mat; however it was found to be less diverse.

Seasonal occurrence and toxicity of Microcystis spp. and Oscillatoria tenuis in the Lebna Dam, Tunisia.

Physicochemical and biological water quality, including the total microcystin concentrations, was investigated for the first time from January to December 2005 in the Lebna Dam, Tunisia. Microcystin levels and characterization of the different microcystin variants present were measured by protein phosphatase (PP2A) inhibition assays and by LC/MS/MS, respectively. Nutrient values were high, with total inorganic nitrogen and phosphorus concentrations ranging from 0.05 to 8.4 mg L(-1) and from 0.03 to 1.37 mg L(-1), respectively. However, the chlorophyll-a concentrations were very low with a peak (5.32 microg L(-1)) on 20 September 2005 at 9 m depth water samples. Microscopic examination of the phytoplankton samples showed the dominance in the autumn of three morphospecies of the genus Microcystis and the species Oscillatoria tenuis. The total (particulate and dissolved) microcystin concentrations at the surface and at 9 m depth water samples ranged between 0.008 and 1.73, and 0.005 and 5.57 microg microcystin (MC)-LR equivalent L(-1), respectively, with a peak on 20 September. The presence of the microcystin synthetase genes (mcyA, -B, and -C) in the lysates of the three morphospecies of the genus Microcystis and the species O. tenuis indicated that these species were responsible for the microcystin production in this system. The analysis of the field cyanobacterial sample extract containing these species by LC/MS/MS revealed the presence of two microcystin variants: microcystin-LR (MC-LR) and microcystin-YR (MC-YR).