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The effects of iron (Fe), zinc (Zn), and molybdenum (Mo) on the biomass yield, lipid content, lipid yield, and fatty acid composition of Chlorella sp. NC-MKM, Graesiella emersonii NC-M1, Scenedesmus acutus NC-M2, and Chlorophyta sp. NC-M5 were studied. Among them, G. emersonii NC-M1 recorded the highest percentage increase in lipid content (140.3%) and neutral lipid (50.9%) under Zn-supplemented condition compared to the control. Also, it showed a 105% and 41.88% increase in lipid yield and neutral lipid under Fe-supplemented condition compared to the control. However, Chlorella sp. NC-MKM recorded an elevation in lipid yield (70.3% rise) and neutral lipid (24.32% rise) compared to the control in Mo-supplemented condition. The enhanced production of reactive oxygen species (ROS) and antioxidant enzyme (SOD and POD) under Fe-, Zn-, and Mo-supplemented condition supports the lipid accumulation. FAME analysis showed that the overall percentage of SFA and MUFA increased after the addition of Fe, Zn, and Mo in a culture medium compared to the control which is vital for a good-quality biodiesel. Further, biodiesel properties derived from FAMEs such as CN, SV, IV, CFPP, OS, υ, ρ, and HHV were found in accordance with biodiesel standard.

In aquatic ecosystems, many phytoplankton species have evolved various inducible defense mechanisms against the predation. The expression of these defenses is affected by environmental conditions such as nutrient availability. Here, we investigated the anti-grazer colony formation in Scenedesmus obliquus at different magnesium concentrations (0-7.3 mg L -1  Mg 2+ ) in the presence of zooplankton (Daphnia)-derived infochemicals. Results showed that at adequate Mg 2+ , S. obliquus formed high proportions of multi-celled (e.g., four- and eight-celled) colonies, resulting in significantly increased number of cells per colony in response to Daphnia filtrate. On the other hand, in Mg 2+ -deficient treatment, the proportion of multi-celled colonies decreased, together with reduced algal growth rate and photosynthetic efficiency. Finally, the treatment without Mg 2+ strongly suppressed the formation of large colony (mainly eight-celled colonies), whereas the algal growth rate was comparable to that in Mg 2+ sufficient treatment. Despite the inhibition of colony formation, the time reaching the maximum number of cells per colony was not affected by the Mg 2+ concentration, which generally took three days in all groups. Our results indicate that Mg 2+  deficient/absent environments significantly reduced anti-grazing colony formation but not the algal growth, suggesting strong dependability of this morphological defensive trait to magnesium fluctuation in S. obliquus.

Polychlorinated diphenyl sulfides and polychlorinated diphenyl ethers are two types of dioxin-like pollutants, which are prevalent in aquatic environments. However, to date, limited information is available regarding their toxicity to green algae. In this study, growth inhibition, effect on pigment content, and oxidative stress potentials of 4,4'-di-CDPS and 4,4'-di-CDE on green algae Scenedesmus obliquus were investigated. The results indicate that the EC 50 values of 4,4'-di-CDPS after 24, 48, 72, and 96 h of exposure were 1.736, 1.172, 0.994, and 0.820 mg/L, while the corresponding values for 4,4'-di-CDE were 0.697, 1.087, 0.833, and 0.327 mg/L. As compared to the control group, most of the measured pigment content in algal cells significantly decreased after 96-h exposure to these two chemicals, suggesting their suppressive capability on the photosynthesis process in algal cells. Additionally, oxidative stress occurred as demonstrated by the significantly inhibited activities of the antioxidant enzymes (total superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxidase (GPx)), and high increases in malondialdehyde (MDA) content in all 4,4'-di-CDE-treated groups and some moderate-dose and high-dose treatments with 4,4'-di-CDPS. Acute toxicity tests and biochemical analysis showed that 4,4'-di-CDE was more toxic than 4,4'-di-CDPS on S. obliquus.

There has recently been an increase in the usage of TiO 2 nanoparticles (NPs). P25 TiO 2 NPs, a mixture of anatase and rutile phase in 3:1 ratio, are generally used for photocatalytic applications because both phases exhibit a synergistic effect on the photocatalytic activity of the TiO 2 NPs. In the present study, increased toxicity of UVA-pre-irradiated P25 TiO 2 NPs on freshwater algae Scenedesmus obliquus was assessed under visible light and dark exposure conditions at actual low concentrations (0.3, 3 and 35 μM of Ti). Photocatalytic property of P25 TiO 2 NPs caused disaggregation of UVA-pre-irradiated NPs, thus significantly decreasing the mean hydrodynamic diameter (MHD) (188.74 ± 0.54 nm) than that of non-irradiated NPs (232.26 ± 0.44). This decrease in diameter of UVA-pre-irradiated NPs may increase its biological activity towards algal samples. All concentrations of pre-irradiated NPs, under both light and dark conditions, showed a significantly lesser cell viability (p < 0.001) when compared with non-irradiated NPs. Increased production of ROS, antioxidant enzymes and lipid peroxidation supported the viability data. Higher exopolysaccharide production and more nuclear damage were observed for pre-irradiated NPs. NP uptake was also more for the pre-irradiated NPs on treated samples when compared with non-irradiated NPs on treated samples, which, in turn, established the higher toxic potential of UVA-pre-irradiated TiO 2 NPs. This study improves our understanding of the toxic effects of UVA-pre-irradiated TiO 2 NPs on freshwater algae, thereby emphasising the need for ecological risk assessments of metal oxide nanoparticles in a natural experimental medium.

Indirect impact of humic acid (HA) on metal accumulation and toxicity (Cd, Ni, Pb, and Hg; 100 μM; 24 h of exposure) in Scenedesmus quadricauda was studied. Algae were pre-cultured on solid (10 and 100 mg HA/L) or in liquid media (1, 5, and 10 mg HA/L) over 30 days and then exposed to metals mentioned above. Accumulation of applied metals irrespective of pre-culture increased in the order Ni < Cd < Pb < Hg. Algae pre-cultured on solid HA-enriched media accumulated more Cd (+ 46% at 10 mg HA/L), Ni (+ 50 and + 81% at 10 and 100 mg HA/L, respectively), and Pb (+ 15% at 100 mg HA/L) but the impact on Hg amount was not detected. Potassium and calcium decreased in response to all metals (K strongly under Hg excess) and HA had negligible impact. Interestingly, fluorescence microscopy detection of reactive oxygen species/nitric oxide (ROS/NO) balance showed that HA pre-culture suppressed ROS signal and stimulated NO signal in response to Cd (indicating positive impact of HA) while ROS signal in Ni and Pb treatments rather increased but NO signal decreased as expected from elevated Ni and Pb accumulation. Hg had clearly the most toxic impact on the ROS/NO balance. Algae pre-cultured in liquid HA-enriched media showed significantly increased Ni accumulation only (+ 14% at a dose 10 mg HA/L). Present study for the first time showed that humic acid may indirectly affect accumulation of metals and that solid HA-enriched medium used for pre-culture is more suitable to increase accumulation of metals by algae.

Gold nanoparticles (GNPs) are widely used for medical purposes, both in diagnostics as well as drug delivery, and hence are prone to release and distribution in the environment. Thus, we have explored the effects of GNPs with two distinct surface capping (citrate and PVP), and three different sizes (16, 27, and 37 nm) at 0.01-, 0.1-, and 1-mg L -1 concentrations on a predominant freshwater alga Scenedesmus obliquus in the sterile freshwater matrix. We have also investigated how an abundant metal ion from freshwater, i.e., Zn 2+ ions may modulate the effects of the selected GNPs (40 nm, citrate, and PVP capped). Preliminary toxicity results revealed that gold nanoparticles were highly toxic in comparison to zinc ions alone. A significant modulation in the toxicity of Zn ions was not noticed in the presence of GNPs. In contrast, zinc ions minimized the toxicity produced by GNPs (both CIT-37 and PVP-37), despite its individual toxicity. Approximately, about 42, 33, and 25% toxicity reduction was noted at 0.05-, 0.5-, and 5-mg L -1 Zn ions, respectively, for CIT-37 GNPs, while 31% (0.05 mg L -1 ), 24% (0.5 mg L -1 ), and 9% (5 mg L -1 ) of toxicity reduction were noted for PVP-37 GNPs. Maximum toxicity reduction was seen at 0.05 mg L -1 of Zn ions. Abbott modeling substantiated antagonistic effects offered by Zn 2+ ions on GNPs. Stability and sedimentation data revealed that the addition of zinc ions gradually induced the aggregation of NPs and in turn significantly reduced the toxicity of GNPs. Thus, the naturally existing ions like Zn 2+ have an ability to modulate the toxicity of GNPs in a real-world environment scenario.

Due to changing global climatic conditions, a lot of attention has been given to cyanobacteria and their bioactive secondary metabolites. These conditions are expected to increase the frequency of cyanobacterial blooms, and consequently, the concentrations of cyanotoxins in aquatic ecosystems. Unfortunately, there are very few studies that address the effects of cyanotoxins on the physiology of phytoplankton species under different environmental conditions. In the present study, we investigated the effect of the cyanotoxin anatoxin-a (ATX-A) on Microcystis aeruginosa (cyanobacteria) and Acutodesmus acuminatus (chlorophyta) under varying light and nitrogen conditions. Low light (LL) and nitrogen limitation (LN) resulted in significant cell density reduction of the two species, while the effect of ATX-A on M. aeruginosa was not significant. However, under normal (NN) and high nitrogen (HN) concentrations, exposure to ATX-A resulted in significantly (p < 0.05) lower cell density of A. acuminatus. Pigment content of M. aeruginosa significantly (p < 0.05) declined in the presence of ATX-A, regardless of the light condition. Under each light condition, exposure to ATX-A caused a reduction in total microcystin (MC) content of M. aeruginosa. The detected MC levels varied as a function of nitrogen and ATX-A concentrations. The production of reactive oxygen species (H 2 O 2 ) and antioxidant enzyme activities of both species were significantly altered by ATX-A under different light and nitrogen conditions. Our results revealed that under different light and nitrogen conditions, the response of M. aeruginosa and A. acuminatus to ATX-A was variable, which demonstrated the need for different endpoints of environmental factors during ecotoxicological investigations.

To estimate the combined effects of elevated CO2 and temperature on microalgae, three typical and worldwide freshwater species, the green alga Scenedesmus acuminatus, the diatom Cyclotella meneghiniana, and the cyanobacterium Microcystis aeruginosa, as well as mixes of these three species were continuously cultured in controlled environment chambers with CO2 at 390 and 1000 ppm and temperatures of 20, 25, and 30 °C. CO2 and temperature significantly affected the production of microalgae. The cell productivity increased under elevated CO2 and temperature. Although the green alga dominated in the mixed culture within all CO2 and temperature conditions, rising temperature and CO2 intensified the competition of the cyanobacterium with other microalgae. CO2 affected the extracellular polymeric substances (EPS) characteristics of the green alga and the cyanobacterium. Elevated CO2 induced the generation of humic substances in the EPS fractions of the green alga, the cyanobacterium, and the mixed culture. The extracellular carbohydrates of the diatom and the extracellular proteins of the cyanobacterium increased with elevated CO2 and temperature, while the extracellular carbohydrates and proteins of the green alga and the mixes increased under elevated CO2 and temperature. There were synergistic effects of CO2 and temperature on the productivity and the EPS of microalgae. Climate change related CO2 and temperature increases will promote autochthonous organic carbon production in aquatic ecosystems and facilitate the proliferation of cyanobacteria, which potentially changes the carbon cycling and undermines the functioning of ecosystems.

The widespread use of detergents increases the concentration of surfactant in lakes and reservoirs. High surfactant loads produces toxicity to algae; however, the influence of the increasing surfactant on the competition between algae is not clear. In this paper, different amounts of linear alkylbenzene sulfonate (LAS) were added to test the effects of LAS on the competition between Microcystis aeruginosa and Scenedesmus obliquus under eutrophic condition. In single culture, the growth of S. obliquus was promoted under lower LAS concentrations (1 and 20 mg L(-1)), but cell density of S. obliquus reduced when treated with higher LAS concentration (100 mg L(-1)). The growth of M. aeruginosa was inhibited markedly with 20 and 100 mg L(-1) LAS. Compared with single culture, the result was opposite in co-cultures and the cell density of S. obliquus increased significantly when treated with LAS of 1, 20, and 100 mg L(-1). The specific growth rates of S. obliquus and M. aeruginosa in both cultures were 0.4-0.5 day(-1) and 0.6-0.7 day(-1), respectively, except that the specific growth rate of M. aeruginosa in both cultures treated with 100 mg L(-1) LAS was about 0.2 day(-1). M. aeruginosa dominated over S. obliquus in the co-culture without LAS, while the competition was completely opposite with the addition of 20 mg L(-1) LAS. The growth of S. obliquus treated with 20 mg L(-1) LAS was not affected significantly in single culture but was promoted by 75 % in co-culture. Moreover, the growth of S. obliquus in co-culture treated with 100 mg L(-1) LAS was promoted by more than 97 %. These results suggested that the increasing LAS would overturn the competition of algae in freshwater ecosystems.

Despite the extensive research, the mechanism of the antimicrobial and biocidal performance of silver nanoparticles has not been unequivocally elucidated yet. Our study was aimed at the investigation of the ability of silver nanoparticles to suppress the growth of three types of algae colonizing the wetted surfaces or submerged objects and the mechanism of their action. Silver nanoparticles exhibited a substantial toxicity towards Chlorococcales Scenedesmus quadricauda, Chlorella vulgaris, and filamentous algae Klebsormidium sp., which correlated with their particle size. The particles had very good stability against agglomeration even in the presence of multivalent cations. The concentration of silver ions in equilibrium with nanoparticles markedly depended on the particle size, achieving about 6 % and as low as about 0.1 % or even less for the particles 5 nm in size and for larger ones (40-70 nm), respectively. Even very limited proportion of small particles together with larger ones could substantially increase concentration of Ag ions in solution. The highest toxicity was found for the 5-nm-sized particles, being the smallest ones in this study. Their toxicity was even higher than that of silver ions at the same silver concentration. When compared as a function of the Ag(+) concentration in equilibrium with 5-nm particles, the toxicity of ions was at least 17 times higher than that obtained by dissolving silver nitrite (if not taking into account the effect of nanoparticles themselves). The mechanism of the toxicity of silver nanoparticles was found complex with an important role played by the adsorption of silver nanoparticles and the ions released from the particles on the cell surface. This mechanism could be described as some sort of synergy between nanoparticles and ions. While our study clearly showed the presence of this synergy, its detailed explanation is experimentally highly demanding, requiring a close cooperation between materials scientists, physical chemists, and biologists.

Predictive toxicology using chemometric tools can be very useful in order to fill the data gaps for ionic liquids (ILs) with limited available experimental toxicity information, in view of their growing industrial uses. Though originally promoted as green chemicals, ILs have now been shown to possess considerable toxicity against different ecological endpoints. Against this background, quantitative structure-activity relationship (QSAR) models have been developed here for the toxicity of ILs against the green algae Scenedesmus vacuolatus using computed descriptors with definite physicochemical meaning. The final models emerged from E-state indices, extended topochemical atom (ETA) indices and quantum topological molecular similarity (QTMS) indices. The developed partial least squares models support the established mechanism of toxicity of ionic liquids in terms of a surfactant action of cations and chaotropic action of anions. The models have been developed within the guidelines of the Organization of Economic Co-operation and Development (OECD) for regulatory QSAR models, and they have been validated both internally and externally using multiple strategies and also tested for applicability domain. A preliminary attempt has also been made, for the first time, to develop interspecies quantitative toxicity-toxicity relationship (QTTR) models for the algal toxicity of ILs with Daphnia toxicity, which should be interesting while predicting toxicity of ILs for an endpoint when the data for the other are available.

Enrichment of waterways with nitrogen (N) and phosphorus (P) has accelerated eutrophication and promoted cyanobacterial blooms worldwide. An understanding of whether cyanobacteria maintain their dominance under accelerated eutrophication will help predict trends and provide rational control measures. A mesocosm experiment was conducted under natural light and temperature conditions in Lake Taihu, China. It revealed that only N added to lake water promoted growth of colonial and filamentous cyanobacteria (Microcystis, Pseudoanabaena and Planktothrix) and single-cell green algae (Cosmarium, Chlorella, and Scenedesmus). Adding P alone promoted neither cyanobacteria nor green algae significantly. N plus P additions promoted cyanobacteria and green algae growth greatly. The higher growth rates of green algae vs. cyanobacteria in N plus P additions resulted in the biomass of green algae exceeding that of cyanobacteria. This indicates that further enrichment with N plus P in eutrophic water will enhance green algae over cyanobacterial dominance. However, it does not mean that eutrophication problems will cease. On the contrary, the risk will increase due to increasing total phytoplankton biomass.

Flubendazole (FLU) and fenbendazole (FEN) belong to benzimidazoles-pharmaceuticals widely used in veterinary and human medicine for the treatment of intestinal parasites as well as for the treatment of systemic worm infections. In recent years, usage of these drugs increased, which resulted in a larger contamination of the environment and possible negative effects on biota. Hence, in our research, we investigated an aquatic ecotoxicity of these pharmaceuticals towards: marine bacteria (Vibrio fischeri), green algae (Scenedesmus vacuolatus), duckweed (Lemna minor) and crustacean (Daphnia magna). Ecotoxicity tests were combined with chemical analysis in order to investigate the actual exposure concentration of the compounds used in the experiment as well as to stability and adsorption studies. As a result, study evaluating sensitivity of different aquatic organisms to these compounds and new ecotoxicological data is presented. The strongest negative impact of FLU and FEN was observed to D. magna.

Effect of nitric oxide donor (sodium nitroprusside, SNP, 500 μM) or hydrogen peroxide scavenger (dithiothreitol, DTT, 500 μM) on cadmium (Cd) or copper (Cu) uptake (150 μM solutions) and toxicity using Scenedesmus quadricauda was studied. Combined treatments (Cd or Cu + DTT or SNP) usually ameliorated metal-induced toxicity at the level of pigments, proteins, and mineral nutrients in comparison with respective metal alone. Viability tests (MTT and TTC) showed the lowest values preferentially in Cu treatments, indicating higher toxicity in comparison with Cd. Cd showed low impact on amino acids while strong Cu-induced depletion was mitigated by DTT and SNP. Amount of ROS and NO showed the most pronounced responses in SNP variants being rather reciprocal than parallel and regulated ascorbate peroxidase activity. Blot gel analyses of hsp70 protein did not reveal extensive changes after given exposure period. Phenols were elevated by DTT alone while all Cu treatments revealed depletion. Total Cu content decreased while total Cd content increased in metal + SNP or metal + DTT. Subsequent experiment using lower Cd, SNP or DTT doses (10 and 100 μM) revealed concentration-dependent impact on Cd uptake. Overall, DTT was found to be more suitable for the amelioration of metal-induced toxicity.

The geometries of 19 polyaromatic hydrocarbons (PAHs) were fully optimized and calculated by a density functional method (B3LYP) with the 3-21G basis set. Various quantum chemical descriptors such as the energy of the highest occupied molecular orbital (E (HOMO)), the energy of the lowest unoccupied molecular orbital (E (LUMO)), the difference in energy between those orbitals (E (GAP)), electronegativity (χ), chemical potential (μ), chemical hardness (η), softness index (S), electrophilicity (ω), and polarizability (α) were employed along with physicochemical descriptors to construct useful quantitative structure-activity relationship (QSAR) models for the photoinduced toxicity of PAHs toward two aquatic species (Daphnia magna and Scenedesmus vacuolatus). E (LUMO), E (HOMO), E (GAP), S, χ, the molar refractivity (MR), and the molecular weight provide valuable information and play a significant role in the assessment of PAH phototoxicity. The resulting models are not expected to be useful per se for making genuine predictions for much larger test sets, but the various results do demonstrate the potential benefits of incorporating quantum-chemical descriptors into QSAR models for predicting the phototoxicity of PAHs.

The phytotoxic effects of the lichen secondary metabolite-usnic acid on cultures of free living alga-Scenedesmus quadricauda (UTEX 76) and aposymbiotically grown lichen photobiont Trebouxia erici (UTEX 911) were assessed. We found a relatively strong inhibition effect of usnic acid on the growth of alga Scenedesmus, accompanied by an increase of cell size, an alteration of assimilation pigment composition, followed by strong degradation of chlorophyll a, a decrease of chlorophyll a fluorescence, and an increase of reactive oxygen species in the cells. The content of soluble proteins remained a stable parameter. Phytotoxicity of usnic acid on cultures of Trebouxia photobiont was significantly lower. Usnic acid in lichens may act as an allochemical that controls the division of photobiont cells, thereby regulating the balance between the photobiont and mycobiont forming thallus. Higher tolerance to usnic acid in Trebouxia cultures may be an adaptation resulting from the long term co-evolution of these algae with fungi that produce secondary metabolites.

Goal, Scope and Background: The article is focused on dioxin, furan, PCB and organochlorine pesticide monitoring in the surface waters of the Central European, protected natural reserve Krivoklatsko, under the UNESCO programme Man and Biosphere. Persistent compounds are presently transported via different means throughout the entire world. This contamination varies significantly between sites. This raises the question of what constitutes the naturally occurring background levels of POPs in natural, unpolluted areas, but which are close to industrialised regions. Information of real background POP contamination can be of high value for risk assessment management of those sites evidently polluted and for the defining of de-contamination limits. Preserved areas should not be seen as isolated regions in which the impacts of human activities and natural factors are either unexpected or overlooked. Every ambient region, even those protected by a law or other means, are still closely connected to neighbouring human developed and impacted areas, and are therefore subject to this anthropogenic contamination. These areas adjacent to natural reserves are sources of diverse substances, via entry of air, water, soil and/or biota. After an extended period of industrial activities, organochlorine pollutants, even those emitted in trace concentrations have reached detectable levels. For future research and for the assessment of environmental changes, present levels of contamination would be of high importance. This work publishes data of the contamination with organochlorine pollutants of this natural region, where biodiversity and ecological functions are of the highest order., Methods: Semipermeable membrane devices (SPMDs) were utilised as the sampling system. SPMDs were deployed in two small creeks and one water reservoir selected in the central part of the Krivoklatsko Natural Reserve, where it could be expected that any possible contamination by POPs would be lowest. The exposed SPMDs were analysed both for chemical contents of POPs and for toxicity properties. The chemical analyses of dibenzodioxins, dibenzo-furans, PCBs and OCPs were analysed by GC/MS/MS on GCQ or PolarisQ (Thermoquest). Toxicity bioassays were performed on the alga Desmodesmus subspicatus, bacteria Vibrio fischeri and crustacean Daphnia magna. All toxicity data were expressed as the effective volume Vtox. Vtox is a toxicity parameter, the determination of which is independent of SPMD deployment time and pre-treatment dilution (unlike, for example, the EC50 of the SPMD extract)., Results: The following chemical parameters were monitored: (1) tetra, penta, hexa and hepta dibenzo-p-dioxins and furans; (2) all those detectable from tri- through deca-polychloriated biphenyls (PCBs) and (3) a group of organochlorine pesticides: hexachlorobenzene and isomers of hexachlorocyclohexane, DDE, DDD and DDT. The concentrations of dioxins and furans on the assessed sites varied from under detection levels up to 7 pg.l(-1); PCBs were detected in a sum concentration up to 2.8 ng.l(-1); and organochlorine pesticides up to 346 pg.l(-1). The responses of bioassays used were very low, with the values obtained for Vtox being under 0.03 l/d., Discussion: Toxicity testing showed no toxicity responses, demonstrating that the system used is in coherence with the ecological status of the assessed sites. Values of Vtox were under the critical value--showing no toxicity. The PCA of chemical analysis data and toxicity responses resulted in no correlations between these two groups of parameters. This demonstrated that the present level of contamination has had no direct adverse effects on the biota., Conclusions: The concentration values of six EPA-listed, toxic dioxins and sums of tetra-hepta dioxins; nine EPA toxic dibenzofurans and the sums of tetra-hepta bibenzofurans are presented together with all tri-deka PCBs and organochlorine pesticides (alfa-, beta-, gama-, delta-HCH, HCB, opDDE, ppDDE, opDDD, ppDDD, opDDT, ppDDT). These values represent possible current regional natural background values of these substances monitored within the Central European region, with no recorded adverse effects on the freshwater ecosystem (up until the present time)., Recommendations and Perspectives: Assessment of dioxins, furans and other organochlorine compounds within natural reserves can be important for the monitoring of human-induced impacts on preserved areas. No systematic monitoring of these substances in areas not directly affected by industry has generally been realised. There is a paucity of data of the presence of any of these substances within natural regions. Further monitoring of contamination of both soil and biota by dioxins and furans in preserve regions is needed and can be used for future monitoring of man-made activities and/or accidents. Semipermeable membrane devices proved to be a very good sampling system for the monitoring of trace concentrations of ambient organochlorine compounds. Toxicity evaluation using the Vtox concept demonstrated that those localities assessed expressed no toxicity.

A 48 h exposure of Scenedesmus sp. to sublethal concentrations of Cu (2.5 and 10 microM) and Zn (5 and 25 microM) caused a concentration-dependent inhibition of growth, photosynthesis, respiration, NO(3)(-) uptake, and nitrate reductase (EC 1.6.6.1) activity, and a reduction in protein, carbohydrate, and photosynthetic-pigment levels with a concomitant increase in intracellular levels of the test metals. After exposure, algal cells were transferred to the basal medium without the excess level of test metals, to study the recovery of various processes. The growth of the test algae had not recovered up to 12 h after transfer to the basal medium, but some physiological parameters such as photosynthesis and respiration recovered within 6 h. The quicker recovery of photosynthesis and respiration might be used as acclimatory responses as they prepare a background for the recovery of other parameters, including growth, of the test alga by generating energy, forming photosynthate, and establishing the usual catabolism to attain normal conditions. Most of the processes recovered completely or almost completely after being stressed with 2.5 microM Cu or 5 microM Zn. However, the maintenance of a relatively high level of Cu and Zn in the cells previously exposed to 10 microM Cu and 25 microM Zn slowed down the recovery of different processes, which did not fully recover even at the end of the experiment after 96 h. The present study demonstrates that a chain of metabolic events, beginning with respiration and photosynthesis and continuing with assimilation and uptake of nutrients and subsequent restoration of other metabolic processes, is involved in the recovery of the algae from Cu and Zn stress. Each studied parameter seems to play an important role in balancing the cellular homeostasis during recovery from metal stress.

The courses of rRNA accumulation, DNA replication, and nuclear division were followed both in the chloroplast and the nucleocytosolic compartments during the cell cycle in synchronized populations of the chlorococcal alga Scenedesmus quadricauda. Control and nalidixic acid-treated cultures were compared. Nalidixic acid (150 mg/L) was added either at the beginning of the cell cycle or consecutively during the cell cycle to subcultures transferred into the dark. If the inhibitor was applied at the beginning of the cell cycle, chloroplast DNA did not replicate and nucleoids did not divide. Chloroplast division, however, was coordinated in a timely fashion with cytokinesis even under conditions of blocked chloroplast DNA replication. While the growth rate was slowed down, the courses of reproductive processes in the nucleocytosolic compartment were not affected and their timing and the number of rounds were coordinated with growth rate as in the control culture. The rate of cytosolic rRNA synthesis was lower but no apparent effect was seen on the amount of rRNA that accumulated during the cell cycle. In contrast, lower levels of chloroplast rRNA were found at the end of the cell cycle compared with the control culture. Experiments in which cells were transferred to the dark during the cell cycle showed that the inhibitor affected none of the reproductive events in the nucleocytosolic compartment. In the chloroplast compartment, DNA replication was inhibited in inhibitor-treated cultures, but was unaffected in controls. The chloroplast nucleoids themselves divided even in the presence of the inhibitor, reducing their DNA content to a level which corresponded to that in freshly formed control daughter cells.

The effect of cadmium on growth processes (accumulation of RNA, proteins and cell volume), cell cycle reproductive events (DNA replication, mitosis, protoplast fission and daughter-cell formation) and the regulatory activity of histone H1 kinases were monitored in synchronized cultures of the chlorococcal alga Scenedesmus quadricauda. Distinct dosage-dependent inhibitory effects of cadmium ions were found in individual growth and reproductive processes. At concentration of about 60 mumol/L CdCl2, the growth processes were slowed down after about half of the cell cycle but the cells grew to the same or larger size than did untreated cells. At higher concentration, the growth became progressively inhibited, being completely blocked above 240 mumol/L. Total RNA accumulation was the most sensitive growth process. Each of the reproductive events was a target for cadmium ions with increasing sensitivity in the following order: DNA replication, mitosis, protoplast fission and daughter cell formation. Throughout the entire experiment, the activity of "mitosis-specific" histone H1 kinases was negligible in the cadmium (60 mumol/L CdCl2) treated cultures, whilst that of the control culture varied, peaking just prior to nuclear divisions. The activity of "growth-associated" histone H1 kinases was not affected by cadmium ions. No effect was found if cadmium was present during the precommitment period. The longer the period in the presence of cadmium, the stronger inhibition of reproductive events.