Inhibition of nitrogen fixation by nitrite inAnabaena variabilis

Addition of nitrite to rapidly growing, nitrogen-fixing filaments ofAnabaena variabilis caused an immediate drop in nitrogenase activity. This was followed by a transient induction of nitrite reductase, recovery of nitrogen fixation and cyanobacterial growth. The experiments with isolated heterocysts and a partially purified nitrogenase preparation from heterocysts showed that nitrite primarily exerted its inhibitory effect by inactivating nitrogenase irreversibly, rather than interfering with photosynthetic energy conservation.Abbreviations ATCC American type culture collection - Chl chlorophyll - FCCP carbonyl cyanide p-trifluoromethoxy phenylhydrazone - Tes 2-{[2 hydroxy-1,1-bis(hydroxymethyl)ethyl] amino} ethane sulfonic acid     

Toxic cyanobacteria in Greek freshwaters

Cyanobacterial scums, collected in 1987 from four Greek freshwater lakes, were examined for their toxicity to mice. Species ofMicrocystis, Oscillatoria, Anabaenopsis, andAnabaena were dominant in the samples. All samples tested had toxic effects on mice after intraperitoneal injection. The lethal dose (LD₅₀) ranged from 40 to 1500 mg cyanobacterial dry weight kg^?1 body weight and gross pathological signs of poisoning were characteristic of cyanobacterial hepatotoxins. The toxicities of the Greek cyanobacterial blooms were similar to those reported for blooms elsewhere in the world, shown to be responsible for the poisoning of wild and domestic animals.     

Chromate (VI) uptake by and interactions with cyanobacteria

Summary The short-term accumulation of chromate by the cyanobacteriaAnabaena variabilis andSynechococcus PCC 6301 has been described as consisting of a rapid and relatively low level of biosorption of chromate to the cell walls; no energy-dependent uptake was detected. This biosorption was dependent on chromate concentration and could be described by a Freundlich adsorption isotherm for both cyanobacterial species studied. Decreasing the external pH increased the chromate accumulation by both species. Over a longer time period with growth it was shown thatA. variabilis was capable of reducing chromate (VI) to chromium (III) and then accumulating the chromium (III).Synechococcus PCC 6301 showed no further interaction with chromate concentrations over the same time period after the initial biosorption.     

Promising microbial consortia for producing biofertilizers for rice fields

Two cyanobacterial cultures from rice paddies of Kyzylorda Provence, Kazakhstan were isolated and characterized: Anabaena variabilis and Nostoc calsicola. Based on these cultures, new consortia of cyanobacteria, microalgae and Azotobacter were developed: ZOB-1 (Anabaena variabilis, Chlorella vulgaris, and Azotobacter sp.) and ZOB-2 (Nostoc calsicola, Chlorella vulgaris, and Azotobacter sp.). High growth rate and photosynthetic activity of microalgae were observed in these consortia. The active consortium ZOB-1 was selected, which improved germination and growth of rice plants. ZOB-1 was recommended as a biostimulator and biofertilizer for crops.     

Physiological evidence indicates microcystin-LR to be a part of quantitative chemical defense system

The functional role of microcystins (MC) is poorly understood. Here, we investigated the effect of pure MC-LR recovered from the freshwater planktonic cyanobacterium Nostoc sp. BHU001 on five closely related cyanobacteria (Nostoc muscorum, Nostoc commune, Anabaena fertilissima, Anabaena doliolum, and Cylinderospermum majus) isolated from different habitats as well as on the producer itself (Nostoc sp. BHU001). MC-LR was found to be a general growth inhibitor active at nanomolar range (25–100 μg L^?1). It inhibited the growth of all cyanobacterial strains in a concentration-dependent manner, except the producer. A. fertilissima was the most sensitive species. MC-LR affected vital metabolic processes such as photosynthesis, respiration, and nitrogen fixation. Nitrogenase activity showed maximum sensitivity, followed by respiration, photosynthesis, and general growth. The photosynthetic electron transport activity was maximally inhibited at PSI, followed by whole chain and PSII activities. Thus, MC-LR is active at multiple sites causing energy constraint to the vital metabolic processes of the target organisms. However, its requirement at high concentration, which is environmentally irrelevant, and lack of quantitative information on the extracellular release of MC-LR suggest that MC-LR has no allelopathic function and could be a part of a quantitative chemical defense system.     

Treatment of mixed domestic–industrial wastewater using cyanobacteria

Alexandria Sanitary Drainage Company (ASDCO), Alexandria, Egypt has two primary treatment plants, the eastern and the western wastewater treatment plants (EWTP and WWTP) that receive mixed domestic–industrial influents and discharge into L. Mariut. The lake is subjected therefore to severe levels of pollution and dominated by members of cyanobacteria that can cope with the high pollution load in the lake water. Isolation and utilization of the locally generated cyanobacterial biomass for remediation processes of highly toxic pollutants offers a very efficient and cheap tool for governmental or private industrial activities in Alexandria and will generate a source of revenue in Egyptian localities. The main objective of the present study was to investigate the biodegradation and biosorption capacity of some potential cyanobacterial species dominating the lake ecosystem toward organic and inorganic contaminants polluting the primary-treated effluents of the EWTP and WWTP. The primary effluents were subjected to biological treatment using three axenic cyanobacterial strains (Anabaena oryzae, Anabaena variabilis and Tolypothrix ceytonica) as batch system for 7 days. Removal efficiencies (RE) of the different contaminants were evaluated and compared. Results confirmed the high efficiencies of the investigated species for the removal of the target contaminants which were species and contaminant-dependent. BOD₅ and COD recorded 89.29 and 73.68% as maximum RE(s) achieved by Anabaena variabilis and Anabaena oryzae, respectively. The highest RE of the TSS recorded 64.37% achieved by Tolypothrix ceytonica, while 38.84% was recorded as the highest TSD RE achieved by Anabaena variabilis. Tolypothrix ceytonica also exhibited the highest RE for FOG recorded 93.75%. Concerning the contaminant metals, Tolypothrix ceytonica showed the highest biosorption capacity where 86.12 and 94.63% RE were achieved for Zn and Cu, respectively. In conclusion, results of the present study confirmed the advantageous potential of using the tested cyanobacterial species for the treatment of contaminated wastewater. Results also clearly showed the quality improvement of the discharged wastewater which in turn will eliminate or at least minimize the expected deterioration of the receiving environment.     

Nitrogen-fixing cyanobacterium <Emphasis Type="Italic">Trichormus variabilis</Emphasis> of the Lake Baikal phytoplankton

A new filamentous cyanobacterial strain BAC 9610 was isolated from the lake Baikal pelagial. Data obtained by light, scanning, and transmission electron microscopy, along with 16S rRNA gene sequence analysis, allowed the bacterium identification as Trichormus variabilis, previously known as Anabaena variabilis. Trichormus is a cyanobacterial genus not presented in the list of Baikal plankton algae; A. variabilis also hasn’t been previously detected in Baikal phytoplankton. T. variabilis nitrogen fixation ability was demonstrated. The gene responsible for nitrogen fixation, nifH, was identified by PCR and was partially sequenced. No hepatotoxin synthesis genes were revealed in the strain.     

Toxicity of mycotoxins produced by four Alternaria species toArtemia salina larvae

22 isolates ofAlternaria alternata, A raphani, A consortiale, andA chartarum were examined for the production of alternariol (AOH), alternariol methyl ether (AME), altenuene (ALT), altertoxin I (ATX I), and tenuazonic acid (TA) on wheat grain and for toxicity of culture extracts toArtemia salina larvae. The total amount of 5 toxins produced under laboratory conditions ranged from 5 mg/kg to 11.112mg/kg. The toxic extracts showed EC50 values in the range of 3.3 to 144.5 mg/mL. There was no correlation between toxicity of extracts toArtemia salina and the amount of mentioned mycotoxins in culture.     

Relationship between Sodium Influx and Salt Tolerance of Nitrogen-Fixing Cyanobacteria

The relationship between sodium uptake and cyanobacterial salt (NaCl) tolerance has been examined in two filamentous, heterocystous, nitrogen-fixing species of Anabaena. During diazotrophic growth at neutral pH of the growth medium, Anabaena sp. strain L-31, a freshwater strain, showed threefold higher uptake of Na⁺ than Anabaena torulosa, a brackish-water strain, and was considerably less salt tolerant (50% lethal dose of NaCl, 55 mM) than the latter (50% lethal dose of NaCl, 170 mM). Alkaline pH or excess K⁺ (>25 mM) in the medium causes membrane depolarization and inhibits Na⁺ influx in both cyanobacteria (S. K. Apte and J. Thomas, Eur. J. Biochem. 154:395-401, 1986). The presence of nitrate or ammonium in the medium caused inhibition of Na⁺ influx accompanied by membrane depolarization. These experimental manipulations affecting Na⁺ uptake demonstrated a good negative correlation between Na⁺ influx and salt tolerance. All treatments which inhibited Na⁺ influx (such as alkaline pH, K⁺ above 25 mM, NO₃⁻, and NH₄⁺), enhanced salt tolerance of not only the brackish-water but also the freshwater cyanobacterium. The results indicate that curtailment of Na⁺ influx, whether inherent or effected by certain environmental factors (e.g., combined nitrogen, alkaline pH), is a major mechanism of salt tolerance in cyanobacteria.     

Growth and survival potentials of immobilized diazotrophic cyanobacterial isolates exposed to common ricefield herbicides

Summary The effect of graded concentrations of four common ricefield herbicides (Arozin, Butachlor, Alachlor, 2,4-D) on diazotrophic growth, macromolecular contents, heterocyst frequency and tolerance potentials of Ca-alginate immobilized diazotrophic cyanobacterial isolates Nostoc punctiforme, N. calcicola, Anabaena variabilis, Gloeocapsasp., Aphanocapsa sp. and laboratory strain N. muscorum ISU (Anabaena ATCC 27893) was studied and compared with free-living cultures. Cyanobacterial isolates showed progressive inhibition of growth with increasing dosage of herbicides in both free and immobilized states. There were significant differences in the relative toxicity of the four herbicides. Arozin proved to be more growth toxic in comparison to Alachlor, Butachlor and 2,4-D. Growth performance of the immobilized cyanobacterial isolates under herbicide stress showed a similar diazotrophic growth pattern to free cells with no difference in lethal and sub-lethal dosages. However, at lethal concentrations of herbicides, the immobilized cells exhibited prolonged survivability of 14–16 days as compared to their free-living counterparts (8–12 days). The decline in growth, macromolecular contents and heterocyst frequency was found to be similar in both the states in graded dosages of herbicides. Of the test organisms, A. variabilis showed maximum natural tolerance towards all the four herbicides tested. Evidently immobilization by Ca-alginate seems to provide protection to the diazotrophic cyanobacterial inoculants to a certain extent against the growth-toxic action of herbicides.     

Taxonomic status ofAnabaena azollae: An overview

Despite the long-standing and widespread use of the symbiotic association between the aquatic fern Azolla and its cyanobacterial symbiontAnabaena azollae to augment nitrogen supplies in rice paddy soils, very little is known about taxonomic aspects of the symbiosis. The two partners normally remain associated throughout vegetative and reproductive development, limiting the opportunities for interchanges. We have used monoclonal antibodies and DNA/DNA hybridization techniques to show that the cyanobacterial partner is not uniform throughout the genus Azolla, and that substantial diversification has occurred. With these procedures it will be possible to characterize genotypes of the cyanobacterium and to monitor experiments aimed at synthesizing new combinations ofAzolla species andAnabaena azollae strains.     

Experimental associations of cyanobacteria and actinomycetes

Associations of cyanobacteria with actinomycetes are not being investigated. The purpose of this study is to investigate the biological aspects of coexistence of the free-living Anabaena variabilis with actinomycetes isolated from apogeotropic roots of Strangeria eriopus and Cycas micholitzii; with the cyanobacterium Oscillatoria terebriformis (Ag.) Elenk. emend., which were isolated from the natural cyanobacterial mat taken from the Kamchatkan thermal spring; and with actinomycetes isolated from the accumulating culture of cyanobacterium. Positive tropism of actinomycete hyphae to cyanobacterial trichomes and that of the cyanobacterium to streptomycetes were observed. Stimulation of growth of O. terebriformis in the associated culture with the streptomycete was recorded. The increase of fixation of nitrogen by A. variabilis and of photosynthetic activity of O. terebriformis in the associated culture with the streptomycete was recorded.     

Temporal and spatial distribution of planktic cyanobacteria in Lake Kastoria, Greece, a shallow, urban lake

The temporal and spatial distribution of planktic cyanobacteria and some environmental parameters were studied in the shallow, urban Lake Kastoria, Greece from June 1996 to June 1997. Water temperature varied from 6–27 °C, pH from 7.5–8.9 and dissolved O₂ concentration from 0.7–12 mg m^-3 10^-3. The mean annual Chl a concentration was 83 mg Chl a m^-3 indicative of the eutrophic-hypertrophic state of the lake. Cyanobacterial biomass ranged from 11–238 g FW m^-3, constituting about 90% of the total phytoplankton biomass throughout the year. Cyanobacterial biomass was non-uniformly distributed both vertically and horizontally from August to November 1996 and resulted mainly from the distribution of Microcystis. Seven cyanobacterial taxa were reported for the first time in Lake Kastoria. Six taxa were dominant: Microcystis aeruginosa, M. flos-aquae, M. novacekii. Limnothrix redekei, Anabaena sp. and Cylindrospermopsis raciborskii. The dominant cyanobacterial taxa can be grouped on the basis of their distribution patterns (1) Microcystis species: maximum biomass occurring at pH > 8, temperature 12–17 °C, depth < 0.2 m; (2) Anabaena sp. and Cylindrospermopsis raciborskii : maximum biomass at temperatures 23–26 °C; (3) Limnothrix redekei : maximum biomass at temperatures 6–27 °C. Usually, non-uniform, vertical distributions of cyanobacterial biomass were associated with the formation of temperature, pH and O₂ gradients. L. redekei was considered to be a key lake organism since it contributed up to 59% of the cyanobacterial biomass. Interestingly, three of the dominant cyanobacteria Microcystis aeruginosa, Anabaena sp. and Cylindrospermopsis raciborskii belong to genera that include toxin-producing species.     

Recombination protein A gene,recA, fromSpirulina platensis IAM-M135

We report the cloning and sequencing of therecA gene fromSpirulina platensis. A genomic library ofSpirulina was constructed in pUC19 and screened by PCR using oligonucleotides corresponding to the conserved amino acid sequences ofAnabaena variabilis andSynechococcus RecA proteins. TheSpirulina recA gene consists of an open reading frame (ORF) of 1095 nucleotides encoding a protein (365 residues) which shares an identity of 79%, 70% and 57% with the RecA proteins ofAnabaena variabilis, Synechococcus andEscherichia coli respectively. TherecA gene is located close to one end of the clonedBglII fragment and has only 53 bp of 5 nucleotides. The isolation of this gene has implications for the development of gene transfer system(s) forSpirulina.     

Surface lectin binding toAnabaena variabilis and to cultured and freshly isolatedAnabaena azollae

Five fluorescein isothiocyanate (FITC)-labeled lectins and Calcofluor white ST were tested for their binding abilities to vegetative cells and heterocysts of culturedAnabaena variabilis (AVA) and toA. azollae from four species ofAzolla; toAnabaena azollae freshly isolated fromAzolla pinnata (AP),A. caroliniana (AC),A. mexicana (AX), andA. filiculoides (AF); and to cultured akinetes ofAnabaena variabilis and four isolates ofA. azollae. Heterocysts of cultured cells of threeAnabaena isolates (APC, ACC, AXC) were most intensively surface-stained with soybean agglutinin fromGlycine max (SBA)-FITC; those of AX and AC were dimly stained with wheat germ agglutinin fromTriticum vulgaris (WGA); and only heterocysts of AX were dimly stained withDolichos biflorus agglutinin (DBA). Akinetes of cultured cells stained only with ConA. Vegetative cells and heterocysts of all four fresh isolates stained with Jack Beam aglutinin fromCanavalia ensiformis (ConA). None of the cell types were stained with either peanut agglutinin fromArachis hypogea (PNA) or Calcofluor.     

Effects of light intensity and temperature on the photosynthetic irradiance response curves and chlorophyll fluorescence in three picocyanobacterial strains of Synechococcus

Chrococcoid cyanobacteria of the genus Synechococcus are the important component of marine and freshwater ecosystems. Picocyanobacteria comprise even 80% of total cyanobacterial biomass and contribute to 50% of total primary cyanobacterial bloom production. Chlorophyll (Chl) fluorescence and photosynthetic light response (P-I) curves are commonly used to characterize photoacclimation of Synechococcus strains. Three brackish, picocyanobacterial strains of Synechococcus (BA-132, BA-124, BA-120) were studied. They were grown under 4 irradiances [10, 55, 100, and 145 μmol(photon) m^?2 s^?1] and at 3 temperatures (15, 22.5, and 30°C). Photosynthetic rate was measured by Clark oxygen electrode, whereas the Chl fluorescence was measured using Pulse Amplitude Modulation fluorometer. Based on P-I, two mechanisms of photoacclimation were recognized in Synechococcus. The maximum value of maximum rate of photosynthesis (P _max) expressed per biomass unit at 10 μmol(photon) m^?2 s^?1 indicated a change in the number of photosynthetic units (PSU). The constant values of initial slope of photosynthetic light response curve (α) and the maximum value of P _max expressed per Chl unit at 145 μmol(photon) m^?2 s^?1 indicated another mechanism, i.e. a change in PSU size. These two mechanisms caused changes in photosynthetic rate and its parameters (compensation point, α, saturation irradiance, dark respiration, P _max) upon the influence of different irradiance and temperature. High irradiance had a negative effect on fluorescence parameters, such as the maximum quantum yield and effective quantum yield of PSII photochemistry (φ_PSII), but it was higher in case of φ_PSII.     

An angiotensin-converting enzyme-inhibitory metabolite with partial structure of microginin in a cyanobacterium <Emphasis Type="Italic">Anabaena fertilissima</Emphasis> CCC597, producing fibrinolytic protease

Whole cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of Anabaena fertilissima CCC597 revealed the presence of a microginin (M _r?=?565 Da), which was purified using ODS C18 solid-phase extraction followed by quaternary methyl ammonium (QMA)-ion exchange and reverse-phase liquid chromatography. Its partial structure was deduced upon LC-MS/MS as β-amino-α-hydroxy-decanoic acid-Ala-MeLeu-Tyr. Microginin-565 inhibited rabbit angiotensin-converting enzyme activity at an apparent IC₅₀?=?9.8 μM.     

Molecular Characterization of a Fungicidal Endoglucanase from the Cyanobacterium Calothrix elenkinii

A gene responsible for fungicidal activity was identified in the cyanobacterial strain Calothrix elenkinii RPC1, which had shown promise as a biocontrol agent. Functional screening of the genomic library revealed fungicidal (against Pythium aphanidermatum) and endoglucanase activities in two clones. Sequencing revealed an open reading frame of 1,044 bp, encoding 348 amino acid residues with a predicted molecular weight of 38 kDa. Analysis of the deduced amino acid sequence of the putative gene (cael1) showed 99% similarity with the β-1,4-endoglucanase from Anabaena laxa RPAN8 and 97% with the glucanase belonging to the peptidase M20 family of Anabaena variabilis and Nostoc sp. PCC7120, respectively. The putative promoters, ribosomal binding sites and a signal peptide of 22 amino acid residues were identified, revealing the secretory nature of the protein. The phylogenetic tree indicated a close relationship of the gene with Bacillus sp. This study is the first to report on the characterization of an endoglucanase in Calothrix sp.     

Use of a rapid bioluminescence assay for detecting cyanobacterial microcystin toxicity

The recent rise in the awareness of the occurrence of toxic cyanobacterial blooms in aquatic environments, with associated human health problems and animal deaths, has increased the need for rapid, reliable and sensitive methods of determining cyanobacterial toxicity. A luminescent bacterial toxicity test was assessed as a complement to the established mouse bioassay. Seventeen samples including pure cyanobacterial microcystin-LR hepatotoxin, laboratory isolates and natural blooms of cyanobacteria were tested and toxicity data compared with mouse LD50 values. Microcystin-LR and all five microcystin-containing cyanobacterial samples, hepatotoxic by mouse test gave EC50 values of less than 0.46 mg/ml in bioluminescence-based Microtox assays. Of 11 samples non-toxic by mouse bioassay, only two gave an EC50 of less than 0.98 mg/ml by bioluminescence assay. It is suggested that the Microtox bioluminescence assay may prove useful in the preliminary screening of cyanobacterial blooms for microcystin-based toxicity.     

Effects of temperature pre-treatment,desiccation and aging on the viability of spores of halophilic blue-green algae

The effects of temperature, desiccation and aging on the viability of spores of Sambhar salt lake blue-green algae, Anabaena fertilissima and Anabaenopsis arnoldii, were studied. Spores of A. arnoldii were found to be more susceptible to temperature variation, desiccation and storage than spores of A. fertilissima. Pre-treatment of spores with higher temperatures, 37° and 47°C, stimulated germination in A. fertilissima. In a sporulated filament, spores which developed first were generally bigger and more resistant to adverse environmental conditions than spores formed later. The differential loss of viability in spores of a filament may be due to certain intrinsic differences in the physiological/ biochemical properties of the spores.