Applicability of the MTT assay for measuring viability of cyanobacteria and algae, specifically for Microcystis aeruginosa (Chroococcales, Cyanobacteria)

Several assay methods were screened for viability assessment in cyanobacteria using Microcystis aeruginosa FACHB 905. Compared with fluorescent diacetate (FDA), Evan's Blue and autofluorescence, the 3-[4,5-dimethylthiazol-2-yl]2,5-diphenyl tetrazolium bromide (MTT) assay, which was based on the ability of viable cells to reduce MTT to formazan, was found to be reliable and was selected for further study. MTT concentration, incubation time and temperature were optimized for M. aeruginosa. Improvements to the sensitivity and reproducibility of the MTT assay included performing it in the dark to reduce the effects of formazan light sensitivity when extracted in DMSO. Another improvement involved collecting viability data by cell by counting rather than colourimetrically, which was concluded from the fact that oxidoreductase activity, responsible for MTT reduction, would elevate or decrease under stress conditions. Half-life of oxidoreductase in dead cell was calculated to be 3 h. The MTT assay was also found to be applicable to other cyanobacteria and diatoms, including field samples, but not for algae belonging to Chlorophyta, Euglenophyta, Pyrrophyta or Chrysophyta. Based on the above results, we proposed an optimized procedure for the MTT method on Microcystis strains. The use of this assay may be of importance to better understand the dynamics of bloom and the fate of Microcystis under natural or disturbed conditions.

Benthic algae of the Gangqu River, Shangrila, China

We surveyed the benthic algae at 32 sites in the Gangqu River system during May 2005. Among the 162 different taxa observed, 88.8% were diatoms. Achnanthes linearis and Achnanthes lanceolata var. elliptica were the dominant species, comprising 17.1% and 14.3% of the total relative abundance, respectively.

The change of gaseous carbon fluxes following the switch of dominant producers from macrophytes to algae in a shallow subtropical lake of China

Successions of lake ecosystems from clear-water, macrophyte-rich conditions into turbid states with abundant phytoplankton have taken place in many shallow lakes in China. However, little is know about the change of carbon fluxes in lakes during such processes. We conducted a case study in Lake Biandantang to investigate the change of carbon fluxes during such a regime shift. Dissolved aquatic carbon and gaseous carbon (methane (CH4) and carbon dioxide (CO2)) across air-water interface in three sites with different vegetation covers and compositions were studied and compared. CH4 emissions from three sites were 0.62 +/- 0.36, 0.70 +/- 0.36, and 1.31 +/- 0.57 mg m(-2) h(-1), respectively. Correlation analysis showed that macrophytes, rather than phytoplankton, directly positively affected CH4 emission. CO2 fluxes of three sites in Lake Biandantang were significantly different, and the average values were 77.8 +/- 20.4, 52.2 +/- 14.1 and 3.6 +/- 26.8 mg m(-2) h(-1), respectively. There were an evident trend that the larger macrophyte biomass, the lower CO2 emissions. Correlation analysis showed that in different sites, dominant plant controlled CO2 flux across air-water interface. In a year cycle, the percents of gaseous carbon release from lake accounting for net primary production were significantly different (from 39.3% to 2.8%), indicating that with the decline of macrophytes and regime shift, the lake will be a larger carbon source to the atmosphere. (c) 2006 Elsevier Ltd. All rights reserved.

Effect of inclusion of blue-green algae meal on growth and accumulation of microcystins in gibel carp (Carassius auratus gibelio)

Six isonitrogenous (crude protein content: 38%) and isoenergetic (gross energy content: 17 kJ g(-1)) diets were formulated to investigate the effects of inclusion of blue-green algae meal on gibel carp (Carassius auratus gibelio). In each diet, 15% of the protein was supplied by fishmeal; the remainder was supplied by soybean meal and blue-green algae meal. Diet 1 was used as control with no blue-green algae meal whereas the content in diets 2-6 was 15.15, 29.79, 44.69, 59.58 and 74.48%, respectively. Each diet was fed to five groups of gibel carp for 12 weeks in a flow-through system. Final body weight and specific growth rate (SGR) of fish fed diet 5 were significantly lower than the control diet (P < 0.05). Mortality of gibel carp increased with increase in algae meal inclusion (P < 0.05), but there was no significant difference between fish fed diets 3-6 (P > 0.05). Feed conversion efficiency (FCE) decreased with the increase in algae meal inclusion (P < 0.05). Fish-fed diet 6 showed the highest feeding rate (P < 0.05), while there were no significant differences among the other groups (P > 0.05). Apparent digestibility coefficient of dry matter, protein, and energy decreased with increasing algae meal inclusion in the diets (P < 0.05). Aspartate aminotransferase (GOT) activity in the liver was not significantly different among groups (P > 0.05). Liver alanine aminotransferase (GPT) activity of fish-fed diets 4, 5 and 6 was significantly lower than the control diet (diet 1; P < 0.05). Microcystins in the muscle, liver, gallbladder, and spleen increased with increasing algae inclusion (P < 0.05).

The algae-lytic ability of bacterium DC10 and the influence of environmental factors on the ability

A lysing-bacterium DC10, isolated from Dianchi Lake of Yunnan Province, was characterized to be Pseudomonas sp. It was able to lyse some algae well, such as Microcystis viridis, Selenastrum capricomutum, and so on. In this study, it was shown that the bacterium lysed the algae by releasing a substance; the best lytic effects were achieved at low temperatures and in the dark. Different concentrations of CaCl2 and NaNO3 influenced the lytic effects; the ability to lyse algae decreased in the following order: pH 4 > pH 9 > pH 7 > pH 5.5. It was significant to develop a special technology with this kind of bacterium for controlling the bloom-forming planktonic microalgae.

Impacts of elevated CO2 concentration on biochemical composition, carbonic anhydrase, and nitrate reductase activity of freshwater green algae

To investigate the biochemical response of freshwater green algae to elevated CO2 concentrations, Chlorella pyrenoidosa Chick and Chlamydomonas reinhardtii Dang cells were cultured at different CO2 concentrations within the range 3-186 μ mol/L and the biochemical composition, carbonic anhydrase (CA), and nitrate reductase activities of the cells were investigated. Chlorophylls (Chl), carotenoids, carbonhydrate, and protein contents were enhanced to varying extents with increasing CO2 concentration from 3-186 μ mol/L. The CO2 enrichment significantly increased the Chl a/Chl b ratio in Chlorella pyrenoidosa, but not in Chlamydomonas reinhardtii. The CO2 concentration had significant effects on CA and nitrate reductase activity. Elevating CO2 concentration to 186 μ mol/L caused a decline in intracellular and extracellullar CA activity. Nitrate reductase activity, under either light or dark conditions, in C. reinhardtii and C. pyrenoidosa was also significantly decreased with CO2 enrichment. From this study, it can be concluded that CO2 enrichment can affect biochemical composition, CA, and nitrate reductase activity, and that the biochemical response was species dependent.

Benthic algae of the Xiangxi River, China

From June 2000 to June 2002, four sites on the mainstem of the Xiangxi River and one site on each of its major tributaries were sampled 16 times each for benthic algae. All total, 223 taxa (most to species and variety levels) were found (193 Bacillariophyta, 20 Chlorophyta, nine Cyanophyta and one Xanthophyta). The diatoms Cocconeis placentula, Achnanthes linearis, and Diatoma vulgare dominated the system, with relative abundance of 33.3%, 18.8%, and 6.4%, respectively. The abundances of all the other taxa were under 5%, and 210 taxa collectively contributed less than 1% of the total abundance. Taxa diversity peaked in winter and reached a minimum in summer. Species richness varied considerably but was not significantly different over time. Maximum algal density occurred in later winter/spring (1.4 x 10(9) ind./m(2)) but was not significantly different from the minimum density, which occurred in August. Chlorophyll a showed similar seasonal fluctuation but also was not significantly different over time. Canonical correspondence analysis demonstrated that water hardness, depth, conductivity, and alkalinity had important influences on variation of epilithic algae in the Xiangxi River system.

Research progress on algae of the microbial crusts in and and semiarid regions

Microbial crusts are attracting much interest in view of their possible uses in environmental conservation and ecological restoration of the and and semiarid regions. Because algae play an irreplaceable important role in the early formation and the strengthening of microbial crusts, they are paid much more attention to than other cryptogams. In this paper, an overview of the current knowledge on the fine structure and development of microbial crust, focusing on the algal biomass, vertical distribution, succession, influential factors on algae, cohesion of soil stabilization, cementing mechanism for soil particles and the microalgal extracellular polymers is given, with particular emphasis on the authors' researches, and some prospects are put forward as well.

Preliminary study on the photoproduction of hydroxyl radicals in aqueous solution with Aldrich humic acid, algae and Fe(III) under high-pressure mercury lamp irradiation

Under a high-pressure mercury lamp (HPML) and using an exposure time of 4 h, the photoproduction of hydroxyl radicals ((OH)-O-.) could be induced in an aqueous solution containing humic acid (HA). Hydroxyl radicals were determined by high-performance liquid chromatography using benzene as a probe. The results showed that (OH)-O-. photoproduction increased from 1.80 to 2.74 muM by increasing the HA concentration from 10 to 40 mg L-1 at an exposure time of 4 h (pH 6.5). Hydroxyl radical photoproduction in aqueous solutions of HA containing algae was greater than that in the aqueous solutions of HA without algae. The photoproduction of (OH)-O-. in the HA solution with Fe(111) was greater than that of the solution without Fe(III) at pH ranging from 4.0 to 8.0. The photoproduction of (OH)-O-. in HA solution with algae with or without Fe(111) under a 250 W HPML was greater than that under a 125 W HPML. The photoproduction of (OH)-O-. in irradiated samples was influenced by the pH. The results showed that HPML exposure for 4 h in the 4-8 pH range led to the highest (OH)-O-. photoproduction at pH 4.0.

The vertical microdistribution of cyanobacteria and green algae within desert crusts and the development of the algal crusts

Substantial amounts of algal crusts were collected from five different desert experimental sites aged 42, 34, 17, 8 and 4 years, respectively, at Shapotou ( China) and analyzed at a 0.1 mm microscale of depth. It was found that the vertical distribution of cyanobacteria and microalgae in the crusts was distinctly laminated into an inorganic-layer (ca. 0.00 - 0.02 mm, with few algae), an algae-dense-layer ( ca. 0.02 - 1.0 mm) and an algae-sparse-layer ( ca. 1.0 - 5.0 mm). It was interesting to note that in all crusts Scytonema javanicum Born et Flah ( or Nostoc sp., cyanobacterium), Desmococcus olivaceus (Pers ex Ach., green alga) Laundon and Microcoleus vaginatus Gom. ( cyanobacterium) dominated at the depth of 0.02 - 0.05, 0.05 - 0.1 and 0.1 - 1.0 mm, respectively, from the surface. Phormidium tenue Gom. ( or Lyngbya cryptovaginatus Schk., cyanobacterium) and Navicula cryptocephala Kutz.( or Hantzschia amphioxys (Ehr.) Grun. and N. cryptocephala together, diatom) dominated at the depth of 1.0 - 3.0 and 3.5 - 4.0 mm, respectively, of the crusts from the 42 and 34 year old sites. It was apparent that in more developed crusts there were more green algae and the niches of Nostoc sp., Chlorella vulgaris Beij., M. vaginatus, N. cryptocephala and fungi were nearer to the surface. If lichens and mosses accounted for less than 41.5% of the crust surface, algal biovolume was bigger when the crust was older, but the opposite was true when the cryptogams other than algae covered more than 70%. In addition to detailed species composition and biovolume, analyses of soil physicochemical properties, micromorphologies and mineral components were also performed. It was found that the concentration of organic matter and nutrients, electric conductivity, silt, clay, secondary minerals were higher and there were more micro-beddings in the older crusts than the less developed ones. Possible mechanisms for the algal vertical microdistribtion at different stages and the impact of soil topography on crust development are discussed. It is concluded that biomethods ( such as fine species distribution and biovolume) were more precise than mineralogical approaches in judging algal crust development and thus could be a better means to measure the potentiality of algal crusts in desert amelioration.

A physico-chemical comparative study on extracellular carbohydrate polymers from five desert algae

Hydrodynamic properties of five newly isolated algal extracellular polysaccharides with putative adhesive properties are described, using a combination of size exclusion chromatography, total or 'multi-angle' laser light scattering and analytical ultracentrifugation. The respective polysaccharides had been extracted from four filamentous cyanobacteria: Microcoleus vaginatus, Scytonema javanicum, Phormidium tenue and Nostoc sp. and a coccoid single-cell green. algae Desmococcus olivaceus that had been separated from desert algal crusts of the Chinese Tegger Desert. SEC/MALLS experiments showed that the saccharides had, diverse-weight average molecular weights ranging from 4000 to 250,000 g/mol and all five showed either bi-modal or tri-modal molecular weight distribution profiles. Use of the Mark-Houwink-Kuhn-Sakurada (MHKS) scaling relationship between sedimentation coefficient and (weight average) molecular weight for the five samples, assuming a homologous conformation series revealed an MHKS b exponent of (0.33 +/- 0.04), suggesting a conformation between that of a stiff rod (b similar to 0.18) and a random coil (b similar to 0.4-0.5), i.e. a 'flexible rod' or 'stiff coil'. (C) 2003 Elsevier Ltd. All rights reserved.

Extracellular carbohydrate polymers from five desert soil algae with different cohesion in the stabilization of fine sand grain

Extracellular polymeric substances (EPS) from four filamentous cyanobacteria Microcoleus vaginatus, Scytonema javanicum, Phormidium tenue and Nostoc sp. and a coccoid single-cell green alga Desmococcus olivaceus that had been separated from desert algal crusts of Tegger desert of China, were investigated for their chemical composition, structure,and physical properties. The EPS contained 7.5-50.3% protein (in polymers ranging from 14 to more than 200 kD, SDS-PAGE) and 16.2-46.5% carbohydrate (110-460 kD, GFC). 6-12 kinds of monosaccharides, including 2-O-methyl rhamnose, 2-O-methyl glucose, and N-acetyl glucosamine were found. The main carbohydrate chains from M. vaginatus and S. javanicum consisted mainly of equal proportion of Man, Gal and Glc, that from P. tenue consisted mainly of arabinose, glucose and rhamnose. Arabinose was present in pyranose form, mainly alpha-L 1 --> 3 linked, with branches on C4 of almost half of the units. Glucose was responsible for the terminal units, in addition of having some units as beta1 --> 3 and some as beta1 --> 4 linked. Rhamnose was mainly 1 --> 3 linked with branches on C2 on half of the units. The carbohydrate polymer from D. olivaceus was composed mainly of beta1 --> 4 linked xylose, galactose and glucose. The galactose part was present both in beta-pyranose and -furanose forms. Arabinose in alpha-L-furanose form was mainly present as 1 --> 2 and 1 --> 2, 5 linked units, rhamnose only as alpha 1 --> 3 and xylose as beta 1 --> 4. The backbone of the polysaccharide from Nostoc sp. was composed of beta-1 --> 4 linked xylose, galactose and glucose. Most of the glucose was branched on position C6, terminal glucose and 2-O-methyl glucose units are also present. The relationship between structure, physical properties and potential biological function is discussed. (C) 2003 Elsevier Ltd. All rights reserved.

Effect of desert soil algae on the stabilization of fine sands

Four filamentous cyanobacteria, Microcoleus vaginatus, Phormidium tenue, Scytonema javanicum (Kutz.) and Nostoc sp., and a single-celled green alga, Desmococcus olivaceus, all isolated from Shapotou (Ningxia Hui Autonomous Region of China), were batch cultured and inoculated onto unconsolidated sand in greenhouse and field experiments. Their ability to reduce wind erosion in sands was quantified by using a wind tunnel laboratory. The major factors related to cohesion of algal crusts, such as biomass, species, species combinations, bioactivity, niche, growth phase of algae, moisture, thickness of the crusts, dust accretion (including dust content and manner of dust added) and other cryptogams (lichens, fungi and mosses) were studied. The best of the five species were M. vaginatus and P. tenue, while the best mix was a blend of 80% M. vaginatus and 5% each of P. tenue, S. javanicum, Nostoc sp. and D. olivaceus. The threshold friction velocity was significantly increased by the presence of all of the cyanobacterial species, while the threshold impact velocity was notably increased only by the filamentous species. Thick crusts were less easily eroded than thin crusts, while biomass was more effective than thickness. Dust was incorporated best into Microcoleus crust when added in small amounts over time, and appeared to increase growth of the cyanobacterium as well as strengthen the cohesion of the crust. Microbial crust cohesion was mainly attributed to algal aggregation, while lichens, fungi and mosses affected more the soil structure and physico-chemical properties.

Distribution of epilithic algae in the Xiangxi River system and their relationships with environmental factors

The spatial pattern of epilithic algae in the Xiangxi River system was studied in relation to several environmental factors by two-way indictor species analysis (TWINSPAN), detrended correspondence analysis (DCA), and canonical correspondence analysis (CCA). Eighty-nine taxa including diatoms, green algae, and blue-green algae were observed. Diatoms were dominant, and Cocconeis placentula, Cymbella minuta, Diatoma vulgare, and Gomphonema angustatum appeared in most of sampling sites. By TWINSPAN and DCA, thirty-one sites were divided into three groups based on composition and relative richness of benthic algae. CCA indicated that SiO2, pH, total phosphorus, Ca2+, velocity, elevation, and Cl- were significant environmental factors affecting the distribution of algae communities. In this minimal subset, SiO2 and pH were the most influential variables.