Cyanobacterium Anabaena Flos-aquae Research Articles

Effective biosorption of uranium from aqueous solution by cyanobacterium Anabaena flos-aquae.

Anabaena flos-aquae, a typical species of cyanobacterial bloom, was employed as a useful biosorbent for uranium removal. Batch experiments were conducted to examine the effects of different parameters on the uranium uptake amount of Anabaena flos-aquae. The maximum adsorption capacity of 196.4mg/g was obtained under the optimized experimental conditions. The calculations of kinetic and thermodynamic results proved the adsorption process was endothermic, chemisorption, and spontaneous. The adsorption of uranium onto Anabaena flos-aquae was better defined by the Langmuir model, which indicated the process was a monolayer sorption. In addition, the characterization of the biosorbent before and after uranium sorption implied that the dominant functional groups participated in the uranium adsorption process were hydroxyl, amino, and carboxyl. In conclusion, the environmentally friendly and biocompatible characteristics of Anabaena flos-aquae suggest that it can be a promising biosorbent for uranium removal.

Use of lactic acid bacteria as a biological agent against the cyanobacterium Anabaena flos-aquae

In the present study, we assessed whether lactic acid bacteria (LAB) with antimicrobial activity could be used to effectively control Anabaena flos-aquae growth. In our screening of cyanobacteriacidal bacteria from 14 LAB strains belonging to 11 species, we selected six candidate strains that could lyse 90 % or more A. flos-aquae cells compared with a control. Of those, Lactobacillus paraplantarum KCTC 5045T had the strongest cyanobacteriacidal activity showing complete lysis of cyanobacterial cells at the initial densities of ≥104 cells mL−1. A host range assay revealed that L. paraplantarum strongly inhibited A. flos-aquae, Anabaena crassa, Stephanodiscus hantzschii, and Peridinium bipes but weakly inhibited or stimulated the growth of Scenedesmus actus, Pediastrum sp., Cyclotella meneghiniana, Coelastrum reticulatum, Chlamydomonas sp., and Microcystis aeruginosa. In the microcosm experiment using natural freshwater including abundant phytoplankton assemblage, moreover, the LAB under inoculation of about 105 cells mL−1 could completely terminate the co-growth of A. flos-aquae, A. crassa, Anabaena circinalis, and other Anabaena spp. (over 104 cells mL−1) without a secondary bloom by another algal and cyanobacterial species. Our observation revealed that the cyanobacteriacidal bacterium releases two or more extracellular compounds possessing cyanobacteriacidal activity without cell-to-cell contact when the host cyanobacterium necessarily exits in surrounding water. Property study of cyanobacteriacidal substance revealed the release of heat stable and hydrophobic chemical substances that do not appear to be protein and peptide compound. Taken together, our results indicate that LAB may be a potential bio-agent for future use in controlling freshwater Anabaena blooms without causing the problem of pathogenicity if there is no secondary bloom by a resistant species to this bacterium.

Aquatic toxicity of the macrolide antibiotic clarithromycin and its metabolites

The human macrolide antibiotic clarithromycin is widespread in surface waters. Our study shows that its major metabolite 14-hydroxy(R)-clarithromycin is found in surface waters in comparable amounts. This metabolite is known to be pharmacologically active. Additionally, clarithromycin is partly metabolised to N-desmethyl-clarithromycin, which has no antimicrobial activity.For clarithromycin, some ecotoxicological studies on aquatic organisms have been published. However, many of them are not conform with the scientific principles as given in the “Technical guidance for deriving environmental quality standards” (TGD-EQS), because numerous studies were poorly documented and the methods did not contain analytical measurements confirming that the exposure concentrations were in the range of±20% of the nominal concentrations.Ecotoxicological effects of clarithromycin and its two metabolites on the zebrafish Danio rerio (embryo test), the microcrustacean Daphnia magna, the aquatic monocotyledonous macrophyte Lemna minor, the freshwater green alga Desmodesmus subspicatus (Chlorophyta) and the cyanobacterium Anabaena flos-aquae were investigated in compliance with the TGD-EQS. Environmental risk assessment was performed using ErC10 values of Anabaena, the species most sensitive to clarithromycin and 14-hydroxy(R)-clarithromycin in our testing. Based oncomparable toxicity and similar concentrations of clarithromycin and its active metabolite 14-hydroxy(R)-clarithromycin in surface waters, an additional multiplication factor of 2 to the assessment factor of 10 on the ErC10 of clarithromycin should be used. Consequently, a freshwater quality standard of 0.130μgL−1 is proposed for clarithromycin as the “lead substance”. Taking this additional multiplication factor of 2 into account, single monitoring of clarithromycin may be sufficient, in order to reduce the number of substances listed for routine monitoring programs.

Energy status and immune system alterations in Elliptio complanata after ingestion of cyanobacteria Anabaena flos-aquae

Cyanobacteria have often been described as nutritionally poor for herbivorous organisms. To gain additional information on the potential impacts of invertebrates feeding on cyanobacteria, we fed Elliptio complanata mussels with two types of algae: Anabaena flos-aquae (cyanobacteria) and Pseudokirchneriella subcapitata (green algae). Physiological parameters were examined at the energy status, immune system and oxidative stress levels. Energy status was examined by following the rate of electron transport activity in mitochondria (a measure of cellular energy expense) and lipid/sugar stores in the visceral mass. The cyanobacteria were not actively producing toxins. Based on the digestive gland index, the mussels fed equally on either regime. However, the energy status in mussels fed A. flos-aquae revealed that the total sugar was lower in the digestive gland, whereas mitochondrial electron transport activity (MET), once corrected against the digestive gland somatic index, showed increased energy expenses. Acetylcholinesterase activity and lipid peroxidation (LPO) were also higher in mussels fed with A. flos-aquae compared with mussels fed with P. subcapitata. LPO was correlated by mitochondrial activity in both the digestive gland and gills, suggesting that oxidative stress resulted from metabolic respiration. Immunocompetence (phagocytic activity, natural killer cell-like activity, haemocyte count and viability) and humoral level of lysozyme were not affected in mussels by the algae or cyanobacteria regime. Moreover, the xenobiotic conjugating enzyme, glutathione S-transferase, hemoprotein oxidase and vitellogenin-like proteins were not affected in mussel organs via ingestion of A. flos-aquae. Our study suggests that ingestion of cyanobacteria leads to increased energy expenses, oxidative stress and increased acetylcholine turnover in mussels.

Gas Vesicles across Kingdoms: A Comparative Solid-State Nuclear Magnetic Resonance Study

The buoyancy organelles of aquatic microorganisms have to meet stringent specifications: allowing gases to equilibrate freely across the proteinaceous shell, preventing the condensation of water vapor inside the hollow cavity and resisting collapse under hydrostatic pressures that vary with column depth. These properties are provided by the 7- to 8-kDa gas vesicle protein A (GvpA), repeats of which form all but small, specialized portions of the shell. Magic angle spinning nuclear magnetic resonance is uniquely capable of providing high-resolution information on the fold and assembly of GvpA. Here we compare results for the gas vesicles of the haloarchaea Halobacterium salinarum with those obtained previously for the cyanobacterium Anabaena flos-aquae. The data suggest that the two organisms follow similar strategies for avoiding water condensation. On the other hand, in its relatively shallow habitat, H. salinarum is able to avoid collapse with a less costly GvpA fold than is adopted by A. flos-aquae.

Effects of cerium on growth and physiological characteristics of Anabaena flosaquae

In the present study, the effects of cerium (Ce) on the growth and physiological changes were investigated in the cyanobacterium Anabaena flosaquae (A. flosaquae) during a 17-day period. The results showed that the content of chlorophyll a (chl-a) and activity of antioxidase (e.g. superoxide dismutase, peroxidase, and catalase) increased with Ce3+ concentration in the range of 0.05 to 0.1 mg/L and the growth of A. flosaquae was stimulated. While at around 5 mg/L, the content of malondiadehyde (MDA) increased significantly but the activity of antioxidase reduced, which resulted in the ruin of antioxidant defense system. Compared to the control (Ce3+-free), the population size of live cells declined significantly. Microcystin-LR (MC-LR), the most common and toxic cyanotoxins produced by A. flosaquae, was detected and the highest content of MC-LR was observed in 10 mg/L Ce3+ treatment. These results implied that the aqueous environment might suffer a more negative ecological impact when exposed to relatively low Ce3+ concentrations (<0.1 mg/L).

Phytoplankton chytridiomycosis: community structure and infectivity of fungal parasites in aquatic ecosystems

Fungal parasitism is recurrent in plankton communities, especially in the form of parasitic chytrids. However, few attempts have been made to study the community structure and activity of parasites at the natural community level. To analyse the dynamics of zoosporic fungal parasites (i.e. chytrids) of phytoplankton, samples were collected from February to December 2007 in two freshwater lakes. Infective chytrids were omnipresent in lakes, with higher diversity of parasites and infected phytoplankton than in previous studies. The abundance and biomass of parasites were significantly higher in the productive Lake Aydat than in the oligomesotrophic Lake Pavin, while the infection prevalence in both lakes were similar and averaged about 20%. The host species composition and their size appeared as critical for chytrid infectivity, the larger hosts being more vulnerable, including pennate diatoms and desmids in both lakes. The highest prevalence (98%) was noted for the autumn bloom of the cyanobacterium Anabaena flosaquae facing the parasite Rhizosiphon crassum in Lake Aydat. Because parasites killed their hosts, this implies that cyanobacterial blooms, and other large size inedible phytoplankton blooms as well, may not totally represent trophic bottlenecks because their zoosporic parasites can release dissolved substrates for microbial processes through host destruction, and provide energetic particles as zoospores for grazers. Overall, we conclude that the parasitism by zoosporic fungi represents an important ecological driving force in the food web dynamics of aquatic ecosystems, and infer general empirical models on chytrid seasonality and trophodynamics in lakes.

Spatial distribution of resting stages (akinetes) of the cyanobacteria Anabaena flos-aquae in sediments and its influence on pelagic populations

Recruitment of pelagic populations of cyanobacteria from littoral bottom sediments is assumed to be one of the key factors for the development of harmful blooms. Therefore, it is necessary to obtain more information on the possible factors influencing the spatial distribution of the resting stages in littoral bottom sediments. To test the hypothesis that akinete abundances in littoral bottom sediments are associated with silt and the presence of higher water plants, we sampled spatial and vertical distribution and seasonal dynamics of akinetes of cyanobacterium Anabaena flos-aquae in a small Siberian reservoir. We found that akinetes accumulate in silt at stations located in macrophyte beds. Two experiments were carried out to test if there is relationship between size of inoculum and size of the pelagic population of A. flos-aquae. The proportion of A. flos-aquae in total phytoplankton biomass depended on the initial abundance of akinetes in bottom sediments. The model based on laboratory experiments gave good qualitative description of the field data. These findings can be useful to predict the location of the main akinete accumulation zones in a water body and plan the removal of sediment to prevent the cyanobacterial blooms.

Solid-State NMR Characterization of Gas Vesicle Structure

Gas vesicles are gas-filled buoyancy organelles with walls that consist almost exclusively of gas vesicle protein A (GvpA). Intact, collapsed gas vesicles from the cyanobacterium Anabaena flos-aquae were studied by solid-state NMR spectroscopy, and most of the GvpA sequence was assigned. Chemical shift analysis indicates a coil-α-β-β-α-coil peptide backbone, consistent with secondary-structure-prediction algorithms, and complementary information about mobility and solvent exposure yields a picture of the overall topology of the vesicle subunit that is consistent with its role in stabilizing an air-water interface.

Use of cyanobacterial gas vesicles as oxygen carriers in cell culture.

The gas vesicles isolated from the cells of filamentous cyanobacterium Anabaena flos-aquae were treated and sterilized with glutaraldehyde and then evaluated for their effectiveness as gas carriers in cell culture. Anchorage-dependent Vero cells were grown in a packed bed of microcarrier beads under the perfusion of Dulbecco's Modified Eagle's Medium with 1% serum. The culture medium supplemented with 1.8% (v/v) gas vesicles was found to support a 30% higher maximum glucose utilization rate than the same medium without gas vesicles. The gas vesicle suspension was confirmed to have no apparent effects on cell metabolism in T-flask cultures. The study results indicated that the gas vesicles, with high oxygen carrying capacity, can be used to increase the oxygen supply in cell culture systems.

Influence of sodium ion on heavy metal-induced inhibition of light-regulated proton efflux and active carbon uptake in the cyanobacterium Anabaena flos-aquae

The light-induced proton efflux and active carbon uptake are inhibited by mercury and cadmium ions in Anabaena flos-aquae. The inhibitory effects of these heavy metal ions are reversed by 40 mM concentration of sodium. Here we report that light-induced proton efflux is sodium-dependent which leads to a characteristic enhancement in the rate of photosynthetic oxygen generation and carbon fixation. A low concentration (10 μM) of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) significantly inhibited the rate of oxygen generation while 10 μM carbonyl cyanide-m-chlorophenylhydrazone (CCCP) completely blocked the oxygen generation activity in the organism. The chlorophyll-a fluorescence yield indicates that little fluorescence quenching occurred in the absence of sodium ion. Increasing the extracellular sodium ion accelerated both the initial rate and the extent of fluorescence quenching. These results support the assumption that metal-induced inhibition of the photosynthetic machinery may be mediated by the movement of protons.

The Effect of Zooplankton on Microalgae Blooming and Water Eutrophication

Data of field experiments conducted in a eutrophic water body were used to assess the effect of zooplankton on microalgae blooming. The experiment was based on the use of mesocosms to isolate the components of zooplankton and ichthyofauna ecosystems. The population Daphnia longispina is shown to reduce the population of cyanobacteria Anabaenaflos-aquae by a factor of 350 as compared with the water body, thus allowing the phase of highly-transparent water to extend to the mid-summer. D. Longispina can reduce the population of Microcystis aeruginosa, which is associated with the second period of the transparent-water phase. The filtration activity of zooplankton is shown to be able to recover the quality of eutrophic water bodies.

Glutaraldehyde treatment of proteinaceous gas vesicles from cyanobacterium Anabaena flos-aquae.

As potential gas microcarriers, gas vesicles (GVs) were isolated from cultures of the filamentous cyanobacterium Anabaena flos-aquae and treated with glutaraldehyde. The effects of glutaraldehyde treatment on the stability of GVs, against elevated temperatures (40-121 degrees C) and protein-stripping agents such as urea and sodium dodecyl sulfate (SDS), were then examined with the pressure collapse curves generated using pressure nephelometry. The treatment was very beneficial to GVs against the exposure to SDS and urea; however, it did not make the evolution-optimized vesicle structure stronger or more temperature-resistant. In the presence of these protein-stripping agents, the treated vesicles had higher median (50%) collapse pressures (by > or =1 atm) than the untreated ones, at both room temperature and 40 degrees C. This increase has been presumably attributed to the cross-linking of the large GvpC protein to the ribbed GvpA shell, thereby resisting the stripping of GvpC that provides the primary mechanical strength to the vesicle wall. The glutaraldehyde treatment also restored the strength of GVs weakened by a 5-week storage in a refrigerator and, therefore, is expected to improve the stability of GVs for long-term storage. GVs could not be autoclaved. If necessary for the intended applications, glutaraldehyde treatment may also serve to chemically sterilize the vesicles, with the glutaraldehyde subsequently removed by dialysis.

Flotation characteristics of cyanobacterium Anabaena flos-aquae for gas vesicle production.

Cyanobacterium Anabaena flos-aquae was cultivated in photobioreactors for production of intracellular gas vesicles (GVs), as potential oxygen microcarriers. Natural flotation of the buoyant culture was investigated as a potential means of cell harvesting, because filtration and centrifugation tended to destroy the vesicles. Best flotation was found with actively growing culture and when conducted in the dark. The flotation-related cell properties, including the specific GV content, vesicle-collapsed filament density, and intracellular carbohydrate content, were measured to understand the phenomena. During the batch culture, the specific GV content remained relatively constant at 370 microL/(g dry cells) but the filament density (ranging 1.02 to 1.08 g/cm3) showed a decrease-then-increase profile. The increase began when the growth slowed down because of the reduced light availability at high cell concentrations. The dark flotation was studied with both actively growing (mu approximately 0.2 day-1) and stationary-phase cultures. The specific GV content of the stationary-phase culture remained relatively constant while that of the growing culture increased slightly. The intracellular carbohydrate content of the growing culture decreased much faster and more significantly, from 57 to 10 mg/(g dry cells) in </= 8 h. The filament density also decreased, apparently parallel to the profiles of carbohydrate content.

Genes Encoding Proteins Homologous to Halobacterial Gvps N, J, K, F &amp; L are Located Downstream ofgvpCin the CyanobacteriumAnabaena flos-aquae

Only two gas vesicle genes have been previously identified in the cyanobacteria, gvpA and gvpC, both of which encode structural gas vesicle proteins. Analysis of the nucleotide sequence immediately downstream of gvpC in the cyanobacterium Anabaena flos-aquae has revealed the presence of 4 ORFs (open reading frames) the products of which share significant homology with a number of the gene products derived from halobacterial gvp gene clusters. In halobacteria the gas vesicle gene clusters consist of 14 genes involved in gas vesicle synthesis and assembly. The product of Anabaena ORF 1, located immediately downstream of gvpC is homologous to halobacterial GvpNs. For the remaining ORFs the predicted gene products show homology to both GvpJ and GvpA for ORF 2, to GvpK and GvpA for ORF 3, and to both GvpF and GvpL for ORF 4.

Direct observation of protein secondary structure in gas vesicles by atomic force microscopy

The protein that forms the gas vesicle in the cyanobacterium Anabaena flos-aquae has been imaged by atomic force microscopy (AFM) under liquid at room temperature. The protein constitutes "ribs" which, stacked together, form the hollow cylindrical tube and conical end caps of the gas vesicle. By operating the microscope in deflection mode, it has been possible to achieve sub-nanometer resolution of the rib structure. The lateral spacing of the ribs was found to be 4.6 +/- 0.1 nm. At higher resolution the ribs are observed to consist of pairs of lines at an angle of approximately 55 degrees to the rib axis, with a repeat distance between each line of 0.57 +/- 0.05 nm along the rib axis. These observed dimensions and periodicities are consistent with those determined from previous x-ray diffraction studies, indicating that the protein is arranged in beta-chains crossing the rib at an angle of 55 degrees to the rib axis. The AFM results confirm the x-ray data and represent the first direct images of a beta-sheet protein secondary structure using this technique. The orientation of the GvpA protein component of the structure and the extent of this protein across the ribs have been established for the first time.

The characterization of two aminopeptidase activities from the cyanobacterium Anabaena flos-aquae

Aminopeptidase activity, indicated by hydrolysis of the synthetic substrate alanine p-itroanilide, was identified in the cyanobacterium Anabaena flos-aquae. On purification, 2 enzymes were separated by gel filtration chromatography, a 188 kDa multimer (AP-I) and a 59 kDa monomeric metalloprotein (AP-II). Their activities against a range of alanine-containing peptides were screened. Both enzymes were capable of removing a variety of N-terminal residues, including proline. Neither removed N-terminal acidic residues. The activity of AP-I appeared to be limited to di- and tri-peptides, while AP-II was capable of hydrolysing (Ala) 5. It was not possible to assign the active-site chemistry of AP-I to one of the known hydrolase subgroups as none of the potential inhibitors tested had a significant inhibitory effect. This is the first reported purification of aminopeptidases from a cyanobacterium.

Anatoxin-a is a potent agonist of the nicotinic acetylcholine receptor of bovine adrenal chromaffin cells

(+)-Anatoxin-a is a neurotoxic alkaloid produced by the cyanobacterium Anabaena flos-aquae. In this study synthetic (±)-anatoxin-a was tested on isolated bovine adrenal chromaffin cells to determine its ability to evoke secretion of endogenous catecholamines through neuronal-type nicotinic receptor activation. Anatoxin-a was found to act as a potent agonist of the secretory response of chromaffin cells with an EC 50 of 1–2 μM, compared with an EC 50 of 4–5 μM for nicotine. The cells responded to anatoxin-a and nicotine with bell-shaped concentration-response curves consistent with desensitisation at concentrations of anatoxin-a greater than 5 μM and of nicotine greater than 20 μM. The secretion of catecholamines stimulated by anatoxin-a was completely inhibited in a non-competitive manner by the nicotinic antagonist mecamylamine with an IC 50 of 0.4–0.5 μM. In the presence of depolarising concentrations of K + (15 or 50 mM), anatoxin-a increased the secretion of catecholamines in a concentration-dependent manner up to the same maximum as that achieved by anatoxin-a alone. It is concluded that anatoxin-a acts as a potent and selective nicotinic agonist, capable of evoking secretion of endogenous catecholamines from chromaffin cells via their neuronal-type nicotinic receptor.

Two unsaturated 9R‐hydroxy fatty acids from the cyanobacteriumAnabaena flos‐aquae f. flos‐aquae

Abstract(9R‐10E,12Z,15Z)‐9‐Hydroxyotadecatrienoic acid and (9R,10E,12Z)‐9‐hydroxyoctadecadienoic acid were isolated from the nitrogen fixing cyanobacteriumAnabaena flosaquae. f. flos‐aquae and characterized as the corresponding methyl esters. This is the first report of the natural occurrence of 9R‐oxygenated fatty acids.

Hg2+ and Cd2+ induced inhibition of light induced proton efflux in the cyanobacteriumAnabaena flos-aquae

Light induced proton efflux in intact cells ofAnabaena flos-aquae is inhibited by the heavy metals Hg2+ and Cd2+. Furthermore, Hg2+ and Cd2+ reduced the14CO2 fixation, oxygen evolution and carbonic anhydrase activity responsible for H+ efflux.