TOXIN EXTRACTION FROM THE BLUE-GREEN ALGA MICROCYSTIS AERUGINOSA BY DIFFERENT EXTRACTION MEDIA

SUMMARY

The extractability of toxin from Microcystis aeruginosa isolate UV-006 by different extraction media such as O, 1N HC1, 2N CH₃COOH, Triton X-100 and water at different pH levels was investigated. The best recovery of toxin was achieved with water at a pH of 10 while the recoveries with the more acidic media tended to be lower.     

LYSIS OF MICROCYSTIS AERUGINOSA (KÜTZ.) BY BDELLOVIBRIO-LIKE BACTERIA1

Cells of Microcystis aeruginosa (Kützing), collected from water-blooms of Lake Varese, were lysed by Bdellovibrio-like bacteria. The cells were lysed only after penetration. The cyanobacteria and lysing bacteria were characterized by a fibrous glycocalyx. Once the host cell was penetrated, the bacteria remained localized mainly between the host cell wall and cytoplasmic membrane, which appeared partially thickened. Cell lysis began by breakdown of cell structures. The cell wall appeared broken at many sites, and in completely lysed cells, was partially interrupted. The lysis of Microcystis by bacteria could be one of the causes of the death of algal blooms.     

STRONG PROBABILITY OF LETHAL TOXICITY IN THE BLUE-GREEN ALGA MICROCYSTIS VIRIDIS LEMMERMANN1

Lethal toxicity (intraperitoneal, mouse) was examined in relation to Species composition of samples containing bloom-forming Microcystis populations from natural waters and correlated with toxicity of laboratory strains of four Microcystis formas and species. Toxicity was not always associated with the presence of M. aeruginosa f . aeruginosa Elenkin. A sample with almost all cells of M. aeruginosa f . aeruginosa showed no toxicity, However samples comprised of a high percentage of M. viridis Lemmermann often showed lethal toxicity. Toxicity tests were done on culture strains M. aeruginosa f aeruginosa, M. aeruginosa f flos-aquae Elenkin , M. viridis and M. wesenbergii Kamárek. All five cultured strains of M. viridis were found to be toxic, while only one out of nine strains of M. aeruginosa f . aeruginosa was toxic. Six strains of M. wesenbergii showed no toxicity, It is recommended that attention should be paid to the occurrences and possibility of toxic bloom of M. viridis from the standpoint of water management and public health .     

论鲢、鳙对微囊藻的消化利用

1980年—1985年7—8月在水温30℃,溶氧8mg/L以上,pH值7—8,光照度2800lx的限制摄食量条件下,鲢、鳙鱼种对微囊藻干物质的消化率分别为29.45±4.37、26.07±2.15％;蛋白质消化率分别为39.20±3.44、33.71±2.93％;脂肪消化率分别为34.03±5.38、30.29±2.16％;碳水化合物消化率分别为36.61±4.15、30.39±2.81％。鱼种对微囊藻日需量分别为其体重的54.44和60.53％。鱼的生长与其生活水体中藻类的初始浓度有密切关系:(1)藻类浓度在0.4—0.8g/L时鲢、鳙分别增重1.55—7.41和0.78～6.21％;(2)藻类浓度在1.0—1.3g/L时鲢、鳙分别减重2.0—5.9和2.2—8.3％。0.8g/L为鲢、鳙滤食微囊藻的最大浓度。     

THE EFFECT OF SUBLETHAL CONCENTRATIONS OF NATURAL SUSPENDED SILT ON ROUTINE OXYGEN CONSUMPTION BY MOZAMBIQUE TILAPIA (OREOCHROMIS MOSSAMBICUS)

Summary

Exposure of adult Oreochromismossambicusto sublethal concentrations (5 to 20 g silt ?^?1) of natural silt from the Phalaborwa Barrage altered routine oxygen consumption by causing increased rates of oxygen consumption. This indicates that sublethal levels of suspended silt result in a significant (p<0.05) increase in oxygen consumption by the exposed fish due to an increase in metabolic rate associated with increased activity and stress. Any species unable to compensate for this increase in energy expenditure over time would not be able to sustain itself.     

SEDIMENTARY IMPRINT OF MICROCYSTIS AERUGINOSA (CYANOBACTERIA) BLOOMS IN GRANGENT RESERVOIR (LOIRE,FRANCE)1

Analysis of a sediment core taken from the Grangent reservoir in 2004 showed the presence of high concentrations of Microcystis aeruginosa Kütz. colonies at the sediment surface (250 colonies · mL sediment^?1) and also at depths of 25–35 cm (2300 colonies·mL sediment^?1) and 70 cm (600 colonies · mL sediment^?1). Measurements of radioactive isotopes (⁷Be, ¹³⁷Cs, and ²⁴¹Am) along with photographic analysis of the core were used to date the deep layers: the layer located at ?30 cm dates from summer 2003, and that located at ?70 cm from 1990 to 1991. The physiological and morphological conditions of those benthic colonies were compared with those of planktonic colonies using several techniques (environmental scanning electron microscopy [ESEM], TEM, DNA markers, cellular esterases, and toxins). The ESEM observations showed that, as these colonies age, peripheral cells disappear, with no cells remaining in the mucilage of the deepest colonies (70 cm), an indication of the survival thresholds of these organisms. In the benthic phase, the physiological conditions (enzyme activity, cell division, and intracellular toxins) and ultrastructure (particularly the gas vesicles) of the cells surviving in the heart of the colony are comparable to those of the planktonic form, with all the potential needed for growth. Maintaining cellular integrity requires a process that can provide sufficient energy and is expressed in the reduced, but still existing, enzymatic activity that we measured, which is equivalent to a quiescent state.     

PROPERTIES OF MICROCYSTIS AERUGINOSA AND M. FLOS-AQUAE (CYANOPHYTA) IN CULTURE: TAXONOMIC IMPLICATIONS1

Cultures were cloned from a sample containing Microcystis aeruginosa, M. flos-aquae and a few morphological intermediates. The M. aeruginosa cultures remained distinct from the M. flos-aquae cultures in (a) cell size, (b) cell aggregation pattern, (c) width of the mucilage surrounding the multicellular colonies, (d) sharpness of the mucilage boundary, (e) efect of 0.1–1.0 μM calcium chloride on the disaggregation of multicellular colonies, (f) frequency of mucilage mutants and (g) colony morphology on agar media. No M. flos-aquae culture produced morphs resembling M. aeruginosa, inconsistent with proposals that M. flos-aquae is a developmental stage or environmentally-induced variant of M. aeruginosa. After longterm cultivation, but not soon after origanal isolation, several M. aeruginosa cultures contained mutants with diminished mucilage production and an altered colony shape.     

A NUMERICAL TAXONOMIC STUDY OF NOSTOC AND ANABAENA1

Thirty characteristics of 14 Nostoc and 10 Anabaena species were analyzed from previously published data. Using standard numerical taxonomic methods, simple matching coefficients were calculated and a phenogram drawn. The analysis revealed that some of the central characteristics of Nostoc are: a punctiforme stage; motile reproductive stage; plant mass with a dull to shiny luster, non-veined surface, and nonfimbriate margin; some spherical vegetative cells; no cylindrical heterocysts; and some spherical, but no cylindrical akinetes. Some of the central characteristics of Anabaena that were revealed are: no punctiforme stage; a motile vegetative stage; plant mass with a shiny luster, veined surface, and fimbriate margin; no spherical vegetative cells; some cylindrical heterocysts; and some cylindrical, but no spherical, akinetes. In general, Anabaena has larger akinetes and vegetative cells than Nostoc. Based on 30 morphological characteristics and the clustering data of the phenogram, keys were constructed for the Nostoc and Anabaena species studied. The data clearly support two separate and distinct, though similar genera and, less sharply, the separation of the 24 species. The more useful characteristics for separation of the species are size and shape of akinetes, vegetative cells, and heterocysts; color and luster of plant mass; veined plant mass surface; margin fimbriate; and shape of plant mass in nature.     

Is there significant cyclic electron flow around photoreaction 1 in cyanobacteria?

Evidence for a cyclic electron flow has been sought by study of the steady-state poise of P700 and rate of photoreaction 1 in three cyanobacteria. Under an actinic light 1 (440 or 680 nm) the rate of photoreaction 1 is limited by the rate of electron supply provided by photoreaction 2 and by all return electron flow from low potential donors such as ferredoxin and NAD(P)H. Plots of p, the steady-state fraction of P700 reduced, versus the reciprocal intensity, 1/I, yield linear segments of slope Ip. From considerations of a simple model the slopes and extrapolated intercepts of the linear segments provide estimates of the rate of return electron flow. Analysis shows that the total return electron flow cannot be large, by one estimate not more than three times the rate of dark respiration. This result leads to a conclusion that cyclic electron flow (and any dependent phosphorylation) is not a significant process in these cyanobacteria at ordinary light intensities.Abbreviations DAD diaminodurene - PMS phenazine methosulfate     

BIOCHEMICAL COMPOSITION AND FATTY ACID CONTENT OF FILAMENTOUS NITROGEN-FIXING CYANOBACTERIA

The biochemical composition and fatty acid content of twelve strains of filamentous, heterocystous, nitrogen-fixing cyanobacteria have been determined. When grown under diazotrophic conditions, protein, carbohydrate, lipid, and nucleic acids comprised 37–52%, 16–38%, 8–13%, and 8–11% of the dry weight, respectively. The presence of a combined nitrogen source resulted in an increase in the protein content of the cells and a decrease in the levels of lipids and carbohydrates, although biomass productivity was not affected significantly. Biochemical composition also changed during culture growth, with the highest levels of proteins and lipids occurring as the culture entered stationary phase, whereas the highest levels of carbohydrate and nucleic acids were found during the exponential phase. Total fatty acid levels in the strains assayed ranged between 3 and 5.7% of the dry weight. With regard to fatty acid composition, all strains showed high levels of polyunsaturated fatty acids (PUFAs) and saturated fatty acids (SAFAs), with values of 24–45% and 31–52% of total fatty acids, respectively, whereas the levels of monounsaturated fatty acids (MUFAs) were in general lower (11– 32%). Palmitic acid (16:0) was the most prevalent SAFA, whereas palmitoleic (16:1n- 7) and oleic acid (18:1n-9) were the most abundant MUFAs in all the strains. Among PUFAs, γ-linolenic acid (GLA, 18:3n-6) was present at high levels (18% of total fatty acids) in Nostoc sp. (Chile) and at lower levels (3.6% of total fatty acids) in Anabaenopsis sp. The presence of GLA has not been previously reported in these genera of cyanobacteria. The rest of the strains exhibited high levels (12–35% of total fatty acids) of α-linolenic acid (ALA, 18:3n-3). Linoleic acid (18:2n-6) was also present at a substantial level in most of the strains. Eicosapentaenoic acid (EPA, 20:5n-3) was also detected in Nostoc sp. (Albufera). Some filamentous nitrogen-fixing cyanobacteria therefore represent potential sources of commercially interesting fatty acids.     

The ultrastructure of Anabaena azollae in Azolla pinnata

The water fern Azolla pinnata R. Br. was collected in Northwestern India and its structure was examined using scanning and transmission electron microscopy. Emphasis was given to the symbiotic cyanobacterium. Anabaena azollae , and to its relationship to the hairs of the leaf cavities. The cyanobacterial filaments are loosely arranged and often adhere to the protruding hairs and the folded cell walls of the cavities. The vegetative cells show a typical bilayered cell wall. Thylakoids are few and evenly dispersed in mature vegetative cells and appear to lack phycobilisomes. Clusters of polyglucoside granules are distinguished between the thylakoids. Thylakoid membranes are often seen forming whirls and lattices. Polyhedral bodies (carboxysomes) appear particularly frequently in younger cells and in the proximity of polyphosphate bodies. Presence of structured granules, often positioned at both sides of the cross-wall of neighbouring vegetative cells, suggest a positive nitrogen balance. A high frequency of heterocysts is noted, while spores are not observed.
The outer cell wall of the unbranched, mostly two-celled, hairs shows frequent invaginations. The cytoplasma of the mature hair contains numerous organelles, and is penetrated by an electron transparent network with blebs and vesieles appearing. The exchange of metabolites between the symbiotic partners is discussed in relation to the structures noted.     

EXTRACELLULAR ENZYMES OF THE MICROCYSTIS AERUGINOSA PCC 7813 STRAIN ARE INHIBITED IN THE PRESENCE OF HYDROQUINONE AND PYROGALLOL,ALLELOCHEMICALS PRODUCED BY AQUATIC PLANTS1

Several cyanobacterial species have a high potential to dominate in marine environments and freshwater reservoirs, and the ecological and physiological reasons for this phenomenon are not understood comprehensively. In this study, the ability of a Microcystis aeruginosa Kütz. strain to produce free dissolved enzymes was documented. We have observed that this highly toxic strain releases alkaline phosphatase, leucine aminopeptidase, and β‐glucosidase into the ambient environment. Additionally, the inhibitory activity of selected phenols produced by aquatic plants on the activity of these enzymes was analyzed. The investigated compounds, pyrogallol and, to a lesser degree, hydroquinone, decreased the activity of extracellular enzymes produced by M. aeruginosa, with leucine aminopeptidase being the most sensitive to the inhibitors. The noncompetitive character of enzymatic inhibition suggests that the polyphenols produced by aquatic plants are able to influence the activity of different extracellular or membrane‐bound enzymes.     

THE NORMAL AND INDUCED OCCURRENCE OF CYANOPHYCIN INCLUSION BODIES IN SEVERAL BLUE-GREEN ALGAE1

Multi-L-arginyl-poly(L-aspartic acid) [arg-poly(asp)], the polypeptide component of the cyanophycin inclusion body, is found in cells of many blue-green algae. Formation of this material can be induced by a variety of treatments including the addition of excess nitrogen-containing compounds, the addition of specific inhibitors of macromolecular synthesis, and the exclusion of sulfur or phosphorus. Knowledge of the conditions that induce the synthesis of this polypeptide has made possible an ultrastructural survey to determine the presence and cellular location of cyanophycin bodies in a variety of cyanobacteria. Data presented show that certain strains of the unicellular genus Synechococcus Nägeli do not contain arg-poly(asp) under environmental conditions that markedly increase the level of such material in other cyanobacteria. In addition many strains show spatial localization of the cyanophycin bodies under normal growth conditions, and moreover the normal pattern is retained even when massive synthesis of arg-poly(asp) is induced. Finally there is no evidence that these inclusion bodies occur in certain beggiatoan gliding bacteria.     

Monolayer Film of Phycobilisome-Thylakoid Membrane Complexes from Spirulina platensis

Monolayer films of phycobilisome-thylakoid membrane complexes isolated from Spirulina platensis were prepared at air/aqueous solution interface by using the Langmuir-Blodgett technique. The film preparation was optimized with 0.5 M phosphate buffer (pH 7.0) as sub-phase at 20 °C. The monolayer was transferred into grids and into mica surface for observing the surface image of the complexes by transmission electron microscopy and atomic force microscope, respectively. The shape of complexes was disk-like with the diameter of about 50 nm and the thickness of about 35 nm. The absorption and fluorescence spectra of the complexes in the monolayer were consistent with those in buffer solution, which suggests that the complexes in the monolayer preserve the basic functional groups of photosynthetic apparatus and can be used as a model to investigate the structural connection and functional association of the light-harvesting antenna with the reaction centres.     

Bioaccumulation of Gold by Filamentous Cyanobacteria Between 25 and 200°C

Plectonema boryanum UTEX 485 was reacted with aqueous AuCl ₄ ^? solutions ( ～ 2 mM Au) at 25 to 100°C for 1 month, and 200°C for one day. Addition of AuCl₄ ^? to cyanobacteria killed the cultures instantly, and Au was precipitated throughout the cells as nanoparticles. Precipitation of octahedral crystal platelets of Au occurred in the aqueous fluid, with particle size increasing with increase in temperature from about 1.5 μ m at 25°C to 10 μ m at 100°C. Addition of AuCl₄ ^? to suspensions of the dead, autoclaved cyanobacteria also precipitated Au from solution, suggesting that the presence of cell degradation products caused instability of AuCl₄ ^? .     

THE DEVELOPMENT AND POTENTIAL ROLES OF CELL‐WALL TRABECULAE IN CAULERPA MEXICANA (CHLOROPHYTA)1

Siphonous plants represent an alternate scheme to the way most macroscopic plants are constructed. They are single, often large (1–2 m), sometimes morphologically complex, multinucleate (coenocytic) cells where the whole of the cytoplasm is a continuum. Caulerpa mexicana Sond. ex Kütz. is a siphonous tropical marine green alga characterized by four morphologically distinct regions and, as with other members of the genus, by the presence of a dense network of anastomosing cylindrical cell wall in growths called trabeculae. Based on the results of this study, we propose several roles for trabeculae: (i) They are structural components, which likely add some small amount of support in compression but add considerable strength in tension. (ii) As extensions of the cell wall and plasma membrane, they act as diffusion channels from the cell exterior to the interior cytoplasm. It is possible that trabeculae also play a role in determining cell shape through developmental positioning and placement patterns, thus facilitating the diverse shapes found in the morphologically distinct regions of Caulerpa sp.     

A NOVEL TECHNIQUE FOR PREPARATION OF AXENIC CULTURES OF SYMBIODINIUM (PYRROPHYTA) THROUGH SELECTIVE DIGESTION BY AMOEBAE1

The marine amoeba Trichosphaerium Am-I-7 was used as a tool for preparing unialgal axenic cultures of nondigestible Symbiodinium and Porphyridium species. The resistance of these unicellular algae to the amoebal digestive enzymes, and the differential digestion of bacteria, protozoans, and other algae, resulted in cleansed cells of Symbiodinium and Porphyridium that remained in the amoebal food vacuoles. During multiple fission, the amoeba evacuated its food vacuoles and released the trapped and intact algae, which were then successfully cultured. This method of cleaning was especially useful with algal species that were sensitive to antibiotics or other germicidal agents.     

杧果果实的扫描电镜观察

果成熟果实呈黄色长卵形是典型的核果。其外果皮呈革质,中果皮呈肉质,内果皮呈骨质表面有纤维。内果皮包被着种子形成一个大而扁的核。种子扁形,种皮很薄,子叶呈多胚或单胚。通过对果果实结构各个层次的扫描电镜观察,为其贮藏保鲜及繁育栽培工作提供一些科学依据。