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[The characteristics of the structural organization of Chlorella cells cultured over 1 year under space flight conditions].

Ultrastructure of Chlorella cells grown in individual colonies for 12 months on board of the orbital station "Mir" was studied. A number of differences in submicroscopic organization of cell organelles of the experimental population was detected in comparison with the control. Besides, the spectrum of ultrastructural rearrangements of cell organelles was broader, then in our previous experiments that lasted from 4.5 to 30 days.

Cell Nucleus↗

[Changes in the concentration of extracellular products and their composition in a synchronous culture of Chlorella pyrenoidosa in the presence of light and an elevated 02 concentration].

An increase of oxygen concentration in a gaseous mixture passed through cell suspensions of Chlorella pyrenoidosa (from 21 to 100%) stimulates the production of extra-cellular organic substances and changes their composition. The cells accumulate in the medium glycolic acid rather than a variety of other substances. The accumulation of glycolic acid depends on the growth stage of the cells. The relation of production of extracellular organic substances by the algal cells to the ratio between the carboxylase and oxgenase activities of ribulose diphosphate carboxylase is discussed.

Chlorella↗

Production of single cell protein from five strains of the microalga Chlorella spp. (Chlorophyta).

Single cell protein (SCP) production by five strains of Chlorella species (M109, M121, M122, M138, M150), isolated from different habitats, was studied under the influence of eight environmental factors. The ranges for biomass production and SCP were 150-260 mg/l, and 12.4-49.7% dry wt, respectively. The ranges of SCP under the influence of temperature, light, nitrate, ammonia, phosphorus, iron, carbonate, and sodium chloride were in the following respective order (% dry wt): 18.4-43.3, 20.5-42.3, 12.4-35.8, 15.7-41.8, 15.8-49.4, 17.4-49.7, 13.8-35.6, and 0.0-37.7. All the environmental factors had a positive effect on SCP of the five strains except that sodium chloride had a negative effect on SCP of M150. According to these results, both biomass and SCP production can be manipulated and optimized for commercial use by varying the environmental factors used here.

Biomass↗

Molecular mechanism of the alternative expression of the Chlorella virus CVK2 chitosanase gene.

A novel viral strategy for expression of two mRNAs from a single gene yielding two products with seemingly different functions (alternative gene expression) has be found. The vChta-1 gene of Chlorella virus CVK2 produced two functional chitosanase proteins with apparently different roles in viral infection: The larger 65-kDa chitosanase assembled into virion and presumably function at the beginning of infection, while the smaller 37-kDa enzyme remains in the host cytoplasm where it most likely aids in the digestion of the host cell wall prior to viral release. These predicted activities are essential for a cycle of viral infection. The dual expression of the vChta-1 gene most likely occurred by read-through into a downstream gene, ORF245.

Chlorella↗

Designing of plant artificial chromosome (PAC) by using the Chlorella smallest chromosome as a model system.

As a model for plant-type chromosomes, we have been characterizing molecular organization of the Chlorella vulgaris C-169 chromosome I. To identify chromosome structural elements including the centromeric region and replication origins, we constructed a chromosome I specific cosmid library and aligned each cosmid clones to generate contigs. So far, more than 80% of the entire chromosome I has been covered. A complete clonal physical reconstitution of chromosome I provides information on the structure and genomic organization of plant genome. We propose our strategy to construct an artificial chromosome by assembling the functional chromosome structural elements identified on Chrorella chromosome I.

Centromere↗

Perinatal influence of Chlorella vulgaris (E-25) on hepatic drug metabolizing enzymes and lipid peroxidation.

This study evaluates the potential for the perinatal transfer of active constituents and/or metabolites of Chlorella vulgaris (E-25) via modulating the hepatic level of drug metabolizing enzymes and lipid peroxidation. E-25 (100, 300 or 500 mg/kg b.w./day) induced a significant increase in the hepatic levels of glutathione S-transferase (GST) and sulfhydryl (-SH) in fetal and neonatal systems after 14 days treatment to gestating or lactating mice. E-25 (500 mg/kg b.w./day)-modulated inhibition of cytochrome b5 (Cyt. b5), cytochrome P-450 (Cyt. P-450) and malondialdehyde (MDA) level was observed in the fetal and neonatal hepatic tissue following the transplacental or translactational exposure. The observed modulation in the levels of hepatic drug metabolizing enzymes and lipid peroxidation suggest the chemopreventive potential of E-25 via perinatal passage of active constituents and/or metabolites.

Animals↗

Biotoxicity of mercury to Chlorella vulgaris as influenced by amino acids.

The toxicity of mercury ion, on Chlorella vulgaris, is largely influenced by amino acids. Five amino acids, namely alanine, asparagine, glutamic acid, cysteine and histidine, were added separately to the medium containing static dose of mercury. Survival (%) of the alga was reduced with the increasing concentrations of mercury. Of these five amino acids, cysteine was found to be the most effective while alanine and glutamic acid were the least effective on reducing the toxic effect of mercury on the alga measured in terms of growth, chlorophyll and protein content. The order of detoxification was Alanine < Glutamate < Asparargine < Histidine < Cysteine. Amino acids from ligands with Hg2+ making it less toxic to the alga and produce an additional source of energy for growth and development.

Amino Acids↗

Protein glycosylation and myristylation in Chlorella virus PBCV-1 and its antigenic variants.

Chlorella virus PBCV-1 particles contain three glycoproteins, the major capsid protein Vp54 and two minor proteins Vp280 and Vp260. The major capsid protein is myristylated as well as glycosylated. Both modifications are in the carboxyl-terminal portion of the protein. A gene which is modified in a PBCV-1 antiserum-resistant mutant was cloned and sequenced. This gene has an open reading frame of 3099 bases and encodes one of the two large virion glycoproteins (Vp260). Vp260 contains 13 tandem repeats of 61 to 65 amino acids. The mutation deletes the equivalent of four of the amino acid repeat sequences and duplicates one of these sequences.

Amino Acid Sequence↗

Large deletions in the genome of Chlorella virus CVK1.

Large (30-45 kbp) deletions were induced in the Chlorella virus CVK1 genome by UV irradiation. Restriction endonuclease maps of the mutant genomes showed that these deletions occurred in a region located from 1.5 kbp to 47 kbp from the left DNA end. The nucleotide sequences determined around the deletion boundaries indicate that the deletion process took place by both homologous and nonhomologous recombinations. In one case, the recombination site was within a region of about 600 bp, consisting of 40 tandem repetitions of a 15-bp sequence element. The deleted region may contain several multigene families. Northern blot analyses with probes including the genes for translational elongation factor 3 and DNA polymerase showed no discernible aberrancy in the gene expression patterns in the mutants. However, at least two protein bands were missing from the mutant virions.

Base Sequence↗

Analysis of 76 kb of the chlorella virus PBCV-1 330-kb genome: map positions 182 to 258.

Analysis of 76 kb of newly sequenced DNA, located between map positions 182 and 258 kb in the 330-kb chlorella virus PBCV-1 genome, revealed 175 open reading frames (ORFs) of 65 codons or longer. One hundred and five of these 175 ORFs were considered major ORFs. Twenty-one of the 105 major ORFs resembled proteins in databases including ribonucleotide reductase small subunit, RNase III, thioredoxin, glutaredoxin, protein disulfide isomerase, deoxynucleoside kinase, frog virus 3 ATPase, Acetobacter cellulose synthase, a bacteriophage encoded endonuclease, and two C-5 cytosine DNA methyltransferases. One of the ORFs was the PBCV-1 major capsid protein. The 105 major ORFs were evenly distributed along the genome. One set of ORFs was separated by 543 nucleotides whereas 75 of the ORFs were separated by fewer than 100 nucleotides. Nineteen of the 175 ORFs resembled other PBCV-1 ORFs, suggesting that they represent either gene duplications or gene families.

Amino Acid Sequence↗

Laboratory evaluation of the toxicity of Perfluorooctane Sulfonate (PFOS) on Selenastrum capricornutum, Chlorella vulgaris, Lemna gibba, Daphnia magna, and Daphnia pulicaria.

Perfluorooctane sulfonate (PFOS) is an anthropogenic compound found in trace amounts in many environmental compartments far from areas of production. This, along with the highly persistent nature of PFOS, presents a concern for possible effects in aquatic ecosystems. The objective of this study was to determine the toxicity of PFOS in representative freshwater organisms. Toxicity testing using standard laboratory protocols was performed on the green algae Selenastrum capricornutum and Chlorella vulgaris, the floating macrophyte Lemna gibba, and the invertebrates Daphnia magna and Daphnia pulicaria. No observable effect concentration (NOEC) values were generated from the most sensitive endpoints for all organisms. Autotroph inhibition of growth NOEC values were 5.3, 8.2, and 6.6 mg/L for S. capricornutum, C. vulgaris, and L. gibba, respectively. The 48-h immobility NOEC values for D. magna and D. pulicaria were 0.8 and 13.6 mg/L, respectively. In comparison to immobility, the 21-day lethality NOEC for D. magna was 5.3 mg/L. Based on effect (immobility) values, the most sensitive of all test organisms was D. magna. The most sensitive organism based on 50% inhibition of growth (IC(50)) was L. gibba, with an IC(50) value of 31.1 mg/L determined from wet weight. This is 4.3 times less than the LC(50) for D. pulicaria, which was 134 mg/L. Significant adverse effects (p < or = 0.05) were observed for all organisms in concentrations >134 mg/L. The results indicate that under laboratory conditions PFOS is acutely toxic to freshwater organisms at concentrations at or near 100 mg/L. Based on known environmental concentrations of PFOS, which occur in the low ng/L to low microg/L range, there is no apparent risk to freshwater systems. However, further work is required to investigate long-term effects in these and other freshwater organisms.

Alkanesulfonic Acids↗

Investigating the potential impacts of chlorophenols on the lake baikal (Siberia, russia) food web by employing daphnia grazing bioassays and a chlorella growth bioassay

A grazing bioassay was employed to assess the impacts of chlorophenols on Daphnia magna and Daphnia pulex. The effects of two chlorophenols, pentachlorophenol (PCP) and 4-chlorophenol, were investigated at concentrations of 0.001, 0.01, and 0.1 mg . L-1 over a 96-h period. All tests were conducted in water from the southern basin of Lake Baikal (Siberia). For D. magna, grazing rates were significantly depressed after exposure to 0.001 mg . L-1 of PCP for 48 h or to 0.01 mg . L-1 of 4-chlorophenol for 96 h. However, neither chemical continued to depress filtering rates as either dose or time increased, thus effective concentrations (EC50s) could not be determined. This prevents the use of this bioassay as a tool for assessing exposure to chlorophenols, but it is still useful in that it provides insight into potential ecological effects. In the case of D. pulex, depressed rates were also found at 0.001 mg . L-1 of PCP after 48 h; due to problems with the control, no conclusions were drawn for the effect of 4-chlorophenol on this species. The growth rates of Daphnia's prey, Chlorella vulgaris, were also investigated in the presence of these chemicals; no observable effects were found at any concentration during the 96-h period, implying that ecosystem effects may be limited to higher trophic levels.

Journal Article↗

Identification of antihypertensive peptides from peptic digest of two microalgae, Chlorella vulgaris and Spirulina platensis.

The peptidic fractions that inhibited angiotensin I-converting enzyme (ACE) were separated from the peptic digests of 2 microalgae, Chlorella vulgaris and Spirulina platensis, by ion exchange chromatography and gel filtration. Oral administration of peptidic fractions into spontaneously hypertensive rats at 200 mg/kg of body weight resulted in marked antihypertensive effects. Further separation of the peptidic fractions by ODS high-performance liquid chromatography furnished the following active peptides: Ile-Val-Val-Glu (inhibitory against ACE with an IC(50) of 315.3 microM), Ala-Phe-Leu (63.8 microM), Phe-Ala-Leu (26.3 microM), Ala-Glu-Leu (57.1 microM), and Val-Val-Pro-Pro-Ala (79.5 microM) from C. vulgaris; Ile-Ala-Glu (34.7 microM), Phe-Ala-Leu, Ala-Glu-Leu, Ile-Ala-Pro-Gly (11.4 microM), and Val-Ala-Phe (35.8 microM) from S. platensis.

Journal Article↗

Effects of different food (Chlorella) concentrations on the chronic toxicity of cadmium to survivorship, growth and reproduction of Echinisca triserialis (Crustacea: Cladocera).

Extraneous factors have been shown to greatly modify pollutant stress. The present study was conducted with the objective of determining whether differences in food levels could modify chronic toxicity of cadmium to the various life-history parameters of the cladoceran Echinisca triserialis. Laboratory experiments were conducted on a sublethal range of cadmium (0, 2.5, 5.0, 10 and 20 microg litre(-1)) on life-history parameters such as survivorship, longevity, life expectancy, age at first reproduction, total fecundity, neonate size, net reproductive rate (R0), generation time (T), intrinsic rate of natural increase (r) and growth of Echinisca triserialis in relation to different food (Chlorella) levels of 0.5 (low), 1.5 (medium) and 4.5 (high) x 10(6) cells ml(-1). Cadmium levels of 10 microg litre(-1) and above, and low food levels, had a profound effect in decreasing the magnitude of all parameters studied. EC50 levels computed for life-history parameters, such as longevity, life expectancy at birth, total fecundity, R0 and T, were in the range of 2 to 21 microg litre(-1) cadmium, and this is indicative of extreme adverse effects on the population dynamics of E. triserialis when exposed to low food levels. At high food levels the EC50 was not achieved for cadmium in the toxicant range studied. The significance of these results is discussed in relation to field conditions.

Journal Article↗

Effects of copper, chromium and nickel on growth, photosynthesis and chlorophyll a synthesis of Chlorella pyrenoidosa 251.

Individual and combined effects of three heavy metals; namely, copper, chromium and nickel, on growth, photosynthesis and chlorophyll a synthesis of the unicellular green alga, Chlorella pyrenoidosa 251, were determined. Within the concentration range of 0.1 to 1.0 mg litre(-1) of the three heavy metals tested on growth, photosynthesis and chlorophyll a synthesis of the alga, the order of toxicity of the three heavy metals was copper > chromium > nickel. The presence of one of the three heavy metals interacted synergistically with the other two heavy metals in various bimetallic combinations on growth, photosynthesis and chlorophyll a synthesis of the alga.

Journal Article↗

Effects of chlornitrofen, a herbicide, on reproduction of Brachionus urceolaris (Rotatoria) through water and food (Chlorella).

Chronic effects of chlornitrofen (CNP) on the reproduction of Brachionus urceolaris (Rotatoria) were investigated by exposure of individuals to CNP from the egg stage, which had been attached to the adult. The survivors of 12 neonates, which had been exposed to CNP100, 70 or 40 microg liter(-1) decreased to 50% at the age of c. 2, 4 and 6 days, respectively, compared to c. 6.5 days for those exposed to 0, 10 and 20 microg liter(-1) CNP. Release of offspring (mostly two individuals per day) started at two days old. At the peak, four days old, a control female produced 8.1+/-0.9 offspring per day compared with 4.5+/-1.3 (mean+/-SD, n=12) at 40 microg liter(-1). The cumulative numbers of offspring produced by a female were 25.8+/-1.2, 24.2+/-2.9, 22.3+/-3.6 and 13.6+/-3.1 (mean+/-SD, n=12) at control, 10, 20 and 40 microg liter(-1) CNP exposure, respectively. The 50% reproductive impairment concentration was calculated to be 37 microg liter(-1). Growth of neonates was barely detectable at 70 microg liter(-1), and the rapid increase in the effect of CNP from 40 to 70 microg liter(-1) was attributed to an increase in tolerance with growth of the neonates. The effects of CNP on reproduction were also tested by CNP exposure through food (CNP-accumulated Chlorella). The 50% reproductive impairment concentration of CNP in the alga was calculated to be c.60 microg g(-1) (wet weight) by the same method used to assess the dissolved CNP, although the effect of CNP which may have been released from the alga to the water could not be estimated precisely.

Journal Article↗

Light-induced changes in the fluorescence yield of chlorophyll a in vivo. II. Chlorella pyrenoidosa.

The long-term fluorescence induction in Chlorella pyrenoidosa consists of a fast rise of the fluorescence yield from the level S (of the first wave transient) to a maximum M, followed by slower decay to a terminal stationary level T. The maximum M is attained within 40 seconds from the onset of illumination while the decay to the terminal level T lasts for several minutes. The fluorescence rise (S --> M) coincides with an increase in the rate of oxygen evolution, which, however, remains constant during the fluorescence decay (M --> T). Poisons of photosynthesis 3, (3,4-dichlorophenyl)-1,1 dimethylurea (DCMU, o-phenathroline) inhibit the fluorescence induction, while uncouplers of photophosphorylation affect the fluorescence time course only when they function at an early stage of the coupling sequence e.g., carbonyl cyanide p-trifluoremethoxy phenylhydrazone, (FCCP, atabrin). Phosphorylation inhibitors affecting only the terminal esterification step (phlorizin) have little effect on the fluorescence kinetics. These results suggest that the fluorescence induction requires the operation of a phosphorylating electron transport and that it is possibly related to the light-induced structural changes which accompany photophosphorylation.

Antimetabolites↗

Light-induced changes in the fluorescence yield of chlorophyll A in vivo. 3. The dip and the peak in the fluorescence transient of Chlorella pyrenoidosa.

The fluorescence transient of Chlorella pyrenoidosa, excited by saturating light absorbed mainly by system II, has a dip D between the peak I at 75 msec and the large peak P at 400 msec (the times depend on light intensity). This dip is observed in aerobic cells and in anaerobic cells where it is prominent. In anaerobic cells, the I-D decline is hastened almost equally by absorption of either 705 or 650 nm background light. In anaerobic cells, supplementary 700 and 710 nm light given during the transient slightly hastens and heightens P. Methyl viologen, an exogenous system I electron acceptor, eliminates P. Results suggest that system I action causes D, and that P is due to reduction of Q (fluorescence quencher) and intersystem intermediates caused by development of a block in oxidation of XH (X being the primary electron acceptor of light reaction I). Mathematical analysis suggests that if only two forms of Q participate beyond I, then system I action is required for D. If three forms participate, then the system Q --> QH --> Q' (see text) may explain D. The Malkin model (14), in its present form, does not allow D.

Chlorophyll↗