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BACKGROUND: Cyanobacteria are prokaryotic organisms with wide morphological and metabolic diversity. By means of photosynthesis, they convert inorganic compounds into biomolecules, which may have commercial interest. In this work, we evaluated 20 cyanobacterial strains regarding their physiological aspects such as growth, photosynthesis and biochemical composition, some of which are revealed here for the first time. The organisms were cultivated in cylindrical photobioreactors (CPBR) for 144 h and the biomass was obtained. The light inside cultures was constant throughout experimental time and maintained at the saturation irradiance (Ik) of each species. Culture pH was maintained within 7.8 and 8.4 by automatic CO2 bubbling. Growth rate, dry biomass, chlorophyll a, carotenoids, phycocyanin, proteins, carbohydrates, lipids, polyhydroxyalkanoate (PHA) and antioxidant activity were determined. RESULTS: The proportionality of the biochemical composition varied among species, as well as the growth rates. Leptolyngbya sp. and Nostoc sp. (CCIBt3249) showed growth rates in the range of 0.7-0.8 d-1, followed by Rhabdorderma sp. (~ 0.6 d-1), and Phormidium sp. (~ 0.5 d-1). High carotenoid content was obtained in Rhabdoderma sp. (4.0 µg mL-1) and phycocyanin in Leptolyngbya sp. (60 µg mL-1). Higher total proteins were found in the genus Geitlerinema (75% DW), carbohydrates in Microcystis navacekii (30% DW) and lipids in Phormidium sp. (15% DW). Furthermore, Aphanocapsa holsatica showed the highest antioxidant activity (65%) and Sphaerocavum brasiliense, Microcystis aeruginosa, Nostoc sp. (CCIBt3249) and A. holsatica higher levels of PHA (~ 2% DW). CONCLUSIONS: This study reports on the biochemical composition of cyanobacteria that can impact the biotechnology of their production, highlighting potential strains with high productivity of specific biomolecules.

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Microalgae require copper (Cu) in trace levels for their growth and metabolism, it is a vital component of certain metalloproteins. Although this element has been widely studied concerning microalgae physiology, the effects of environmentally relevant levels have been less studied. We studied the photosynthesis and growth of the Chlorophyte Monoraphidium sp. exposed to Cu ranging from low (1.7 nM) to high (589.0 nM) free Cu ions (Cu2+) concentrations. The growth rate was unaffected by Cu concentrations in the range of 1.7-7.4 nM Cu2+, but decreased beyond it. The relative maximum electron transport rate (rETRm), saturation irradiance (Ek), photochemical quenching (qP and qL), and PSII operating efficiency [Formula: see text] were stimulated in the 3.4-7.4 nM Cu2+ range, concentrations slightly higher than the control, whereas non-photochemical quenching (NPQ) gradually increased with increasing Cu2+. The photosystem II antenna size [Sigma (II)440] increased under high Cu (589.0 nM), which resulted in a decrease in the quinone A (QA) reduction time (tau). In contrast, the QA re-oxidation time was unaffected by Cu exposure. These findings show that a slight increase in Cu stimulated photosynthesis in Monoraphidium sp., whereas high Cu reduced photosynthesis and increased the dissipation of captured light energy. This research is a contribution to the understanding of the dynamic photo-physiological responses of Monoraphidium sp. to Cu ions.

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Atrazine is a herbicide widely used in the control of weeds in crops such as corn, sugar cane, and sorghum. It is often found in aquatic environments, where it can potentially endanger nontarget organisms such as microalgae. The present study evaluated atrazine toxicity to seven different species of Chlorophyceae and the tolerance of the species to the herbicide was related to morphological, photosynthetic, chlorophyll-a content and the activity of the glutathione-S-transferase enzyme (GST). The comparison of median effect concentration (EC50) values for growth inhibition indicates higher toxicity of atrazine for Pseudopediastrum boryanum and Desmodesmus communis, intermediate toxicity for Ankistrodesmus densus, Chlamydomonas puliminiorfes, and Raphidocelis subcapitata, and lower toxicity for Kirchneriella lunaris and Ankistrodesmus falcatus (EC50: 38, 42, 66, 103, 248, 1004, and 1585 µg L-1 atrazine, respectively). Principal component analysis (PCA) with algal characteristics suggested that the atrazine-sensitive algae P. boryanum and D. communis were positively associated with photosynthetic levels and negatively associated with GST activity and chlorophyll-a concentration. The PCA also suggested that the atrazine-tolerant algae A. falcatus and K. lunaris were positively associated with morphological parameters, where the larger the cell size, the more tolerant. Although it is difficult to associate a single characteristic of algae as the key factor determining the tolerance to atrazine, results presented in this work indicate that the cell area, the photosynthetic parameters (mainly saturating irradiance), chlorophyll-a content, and the biotransformation by GST in combination may be potential predictors for the differential tolerance of Chlorophyceae species to the herbicide. Environ Toxicol Chem 2022;41:1675-1685. © 2022 SETAC.

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In order to contribute to the biotechnology of microalgae cultivated in vinasse, we carried out the bioprospection of tolerant species and synthesized biomolecules of the total biomass (microalgae and bacteria), recovered from cultures. To use vinasse as a culture medium for the microalgae, waste was centrifuged and used in concentrations from 5 to 50%. Daily cell densities, growth rates, and EC50 values were obtained. After defining the best pair of vinasse concentration/microalgae strain, dry biomass, and composition (proteins and carbohydrates) were determined in 96 h cultures, considering the associated community (bacteria and yeast). The microalgae tested were Chlamydomonas sp., Chlorella sorokiniana, Chlorella vulgaris, Desmodesmus spinosus, Haematococcus pluvialis, Monoraphidium sp., Scenedesmus quadricauda, and Tetraselmis gracilis. The results showed that although the microalgal growth rates in vinasse were similar to controls in BG11, the cells in vinasse had higher biovolumes, dry biomass, and total proteins. The species H. pluvialis, S. quadricauda, and T. gracilis showed the best productivity parameters in vinasse, despite lower growth rates than the other species. Using low concentrations of centrifuged vinasse as a culture medium, only 22% of biological contaminants were present, thus most of the processed biomass was mainly composed of microalgae. Thus, Chlamydomonas sp., D. spinosus, S. quadricauda, and H. pluvialis microalgae have attributes such as resistance and biomolecules that make them candidates for further optimization in production systems, combining the environmental benefits of using waste with the production of biomolecules and/or biomass of commercial interest.

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ABSTRACT The cultivation of microalgae is a possible destination for vinasse, a residue from the sugar and alcohol industry. This use can help reduce the costs of microalgae production and remediate this residue rich in nutrients. However, the physicochemical characteristics of vinasse limit its use for microalgae growth at low concentrations, except when the residue is pretreated. This work aimed at optimizing the vinasse pretreatments of centrifugation and adsorption by smectite clay and activated charcoal on laboratory scale in terms of amounts of materials used and time spent, making them more viable on larger scales. The optimized processes were then compared in productive, economic, and environmental terms. The dilution of treated vinasse with distilled water resulted in similar growth of Chlorella vulgaris to those obtained with the dilution in BG11 medium, indicating that the addition of nutrients in culture media is not necessary. Although microalgae growth occurs in higher concentrations of vinasse treated by adsorption, the results show that centrifugation required less processing time, has cheaper processing costs, and generated much less residue. Centrifugation treatment has greater economic and environmental viabilities and was more sustainable than the adsorption, even though the algae did not grow in the centrifuged residue in concentrations as high as it did after the adsorption treatment. Therefore, this article brings a new view about the economic and environmental aspects on the use of pretreated vinasse for microalgal growth, giving a lucrative destination for a highly polluting waste.

RESUMO O cultivo de microalgas é um possível destino para a vinhaça, um resíduo da indústria sucroalcooleira. Esse uso pode ajudar a reduzir custos da produção de microalgas, além de remediar esse resíduo rico em nutrientes, entretanto as características físico-químicas da vinhaça limitam seu uso para o crescimento de microalgas a baixas concentrações, exceto quando o resíduo é pré-tratado. Este trabalho objetivou otimizar os pré-tratamentos da vinhaça de centrifugação e de adsorção por argila esmectita e carvão ativado em escala laboratorial quanto à quantidade de materiais utilizados e tempo gasto, tornando-os mais viáveis em escalas ampliadas. Os processos otimizados foram então comparados em termos produtivos, econômicos e ambientais. A diluição das vinhaças tratadas com água destilada resultou em crescimento de Chlorella vulgaris semelhante ao obtido com diluições com meio BG11, indicando que a adição de nutrientes nos meios de cultura não é necessária. Embora o crescimento de microalgas ocorra em concentrações mais altas de vinhaça tratada por adsorção, os resultados mostram que a centrifugação exigiu menos tempo de processamento, tem custos mais baratos e gerou menos resíduos. De maneira comparativa, o tratamento por centrifugação possui maior viabilidade econômica e ambiental e foi mais sustentável do que a adsorção, embora as algas não tenham crescido no resíduo centrifugado em concentrações tão altas quanto após o tratamento de adsorção. Portanto, este artigo traz uma nova visão sobre os aspectos econômicos e ambientais do uso da vinhaça pré-tratada para o crescimento de microalgas, proporcionando um destino lucrativo para um resíduo altamente poluente.

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Vinasse is a waste from the alcohol industry that can support microalgal growth because of its mineral and organic contents. Growing microalgae on waste can be a means of its remediation and decrease culture costs. The physiological knowledge of microalgae on waste can generate subsidies to increase biomass productivity. However, few studies have explored the photosynthetic metabolism of microalgae on complex matrices such as vinasse. In the present research, we investigated some physiological aspects related to photosynthesis and growth of several microalgae in centrifuged and diluted vinasse. The species investigated were Chlamydomonas sp., Chlorella sorokiniana, Chlorella vulgaris, Desmodesmus spinosus, Haematococcus pluvialis, Monoraphidium sp., Scenedesmus quadricauda and Tetraselmis gracilis. Daily cell density, in vivo chlorophyll a and final cellular biovolumes (96 h) were monitored to infer about population growth. Maximum and operational photosynthetic yields (ϕM and ϕ'M), photochemical (qP) and non-photochemical (NPQ) quenchings, light saturation curves and related parameters (α, rETRm and EK), as well as theoretical carbon fixation were evaluated using pulse amplitude modulated fluorometry (PAM). The results showed that chlorophyll a, photochemical (qP) and non-photochemical (NPQ) quenchings were not affected in vinasse, but photosynthetic yields, light curves parameters and autotrophic carbon fixation decreased. Connecting these results to the increase in productivities in vinasse lead us to the rationale that mixotrophic metabolism was supporting microalgae growth on the waste. This study offers important insights into the understanding of microalgal photosynthetic physiology on complex organic wastes, generating subsidies for optimization of biotechnological use of vinasse related to microalgae production.

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Copper (Cu) nanomaterials have been increasingly researched and produced for many different consumer products. They have high reactivity and bactericidal properties, making them important in antifouling paints, which are thus directly introduced into aquatic ecosystems. However, studies are scarce on the behavior of Cu nanoparticles (Cu-NPs) in natural aquatic systems and their interactions with primary producers such as microalgae. We investigated the effects of NPs on some physiological responses of the freshwater phytoplankton Chlorella sorokiniana. The cells were exposed to nominal concentrations ranging from 2.50 to 635.00 µg L-1 Cu-NPs for 96 h under laboratory-controlled conditions. The cultures were monitored daily for population growth and maximum photosynthetic quantum yield. Total lipids, proteins, and carbohydrates were quantified at 72 h of Cu-NP exposure. The results showed a positive correlation between nominal Cu-NPs and Cu in the biomass (0.97 correlation coefficient) and that this was inversely proportional to total carbohydrates, with a -0.64 correlation coefficient. At the higher end of the Cu-NP concentrations tested, higher total proteins and reduced growth rates were obtained in comparison with controls; we suggest that metal-binding proteins/antioxidants and nonstructural proteins were preferentially produced under these conditions. Our results contribute to an understanding of the interaction between Cu-NPs and a cosmopolitan phytoplankton, C. sorokiniana, and we emphasize that the disposal and use of Cu-NPs requires monitoring because even at environmentally relevant concentrations, the composition of the algae was affected. Environ Toxicol Chem 2019;38:387-395. © 2018 SETAC.

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Mixotrophy is a metabolic strategy in which an organism is autotrophic and heterotrophic simultaneously. Considering that the aquatic environment provides several organic sources of carbon, it is probably common for microalgae to perform mixotrophy and not only photoautotrophy, but little is known about microalgae mixotrophy. The present work aimed at investigating the growth, photosynthetic activity, morphology, and biochemical composition of the microalga Chlorella sorokiniana in mixotrophic and photo-mixotrophic conditions, comparing it with photoautotrophy. The results showed pH changes after glucose addition, reaching pH 11.62 in mixotrophic and 10.47 in sequential photo-mixotrophic cultures, which limited the microalgal growth. Highest biomass was obtained in the mixotrophic culture in comparison with the sequential photo-mixotrophic one. Rapid light saturation curves showed that α (photosynthetic efficiency, 1.69) and relative electron transport rate (rETR; 565.61) were higher in the mixotrophic cultures, whereas the highest Ik (irradiance saturation, 386.68) was obtained in the photoautotrophic ones. In the sequential photo-mixotrophic cultures, photosynthetic activity varied during glucose consumption, decreasing the maximum quantum yield Fv/Fm after glucose addition, indicating change in metabolism, from photoautotrophy to mixotrophy by the microalga. The results showed that the mixotrophic cultures had higher production of chlorophyll a (6.26 mg mL-1), cell density (6.62 × 107 cell mL-1), and lipids (0.06 pg µm-3). Sequential photo-mixotrophic cultures showed the highest biovolume (360.5 µm3 cell-1) and total carbohydrates (0.026 pg µm-3). The protein concentration was 3.2 and 2.4 times higher in photoautotrophy and photo-mixotrophic growth, respectively, than in mixotrophy, but lipids were three times higher under mixotrophy. The biochemical changes we observed indicate that the microalga's plasticity in face of new environmental characteristics, such as the presence of organic carbon, can change the flow of energy through natural ecosystems.

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Annona glabra L. is a semi-deciduous tree that contains several active substances, including secondary metabolites, with antifungal activity. Phytopathogenic strains of the genus Pythium cause billion dollar losses all over the world on natural and crop species. Searching for eco-friendly algaecides and fungicides, we analyzed the effects of acetone extracts of A. glabra leaves on the algae Rhaphidocelis subcapitata (Korshikov) and on the oomycete Pythium aphanidermatum (Edson). We evaluated ten extract concentrations for each organism - 0 to 400 mg L-1 for algae and 0-1000 µg disc1 for oomycete. The results showed no effect on algae up to 75 mg L-1, but a significant inhibitory effect at 125 mg L-1 and above, which reduced the growth rate and the final biomass of the algae. Extract concentrations above 200 mg L-1 were completely inhibitory. The half maximal inhibitory concentration for 72 and 96 h of exposure to our crude extracts are comparable to those obtained with commercial fungicides and herbicides used in aquatic ecosystems. The P. aphanidermatum inhibition concentrations have effects comparable to fungicides as Cycloheximide and Bifonazole. Some substances isolated from the extracts are described as antifungals, which could explain part of anti-oomycete activity. Our results highlight the importance of searching bioactive compounds from plants.

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ABSTRACT Annona glabra L. is a semi-deciduous tree that contains several active substances, including secondary metabolites, with antifungal activity. Phytopathogenic strains of the genus Pythium cause billion dollar losses all over the world on natural and crop species. Searching for eco-friendly algaecides and fungicides, we analyzed the effects of acetone extracts of A. glabra leaves on the algae Rhaphidocelis subcapitata (Korshikov) and on the oomycete Pythium aphanidermatum (Edson). We evaluated ten extract concentrations for each organism - 0 to 400 mg L-1 for algae and 0-1000 µg disc1 for oomycete. The results showed no effect on algae up to 75 mg L-1, but a significant inhibitory effect at 125 mg L-1 and above, which reduced the growth rate and the final biomass of the algae. Extract concentrations above 200 mg L-1 were completely inhibitory. The half maximal inhibitory concentration for 72 and 96 h of exposure to our crude extracts are comparable to those obtained with commercial fungicides and herbicides used in aquatic ecosystems. The P. aphanidermatum inhibition concentrations have effects comparable to fungicides as Cycloheximide and Bifonazole. Some substances isolated from the extracts are described as antifungals, which could explain part of anti-oomycete activity. Our results highlight the importance of searching bioactive compounds from plants.

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This research aimed at evaluating cadmium (Cd), copper (Cu), lead (Pb), and zinc (Zn) speciation in water samples as well as determining water quality parameters (alkalinity, chlorophyll a, chloride, conductivity, dissolved organic carbon, dissolved oxygen, inorganic carbon, nitrate, pH, total suspended solids, and water temperature) in a eutrophic reservoir. This was performed through calculation of free metal ions using the chemical equilibrium software MINEQL+ 4.61, determination of labile, dissolved, and total metal concentrations via differential pulse anodic stripping voltammetry, and determination of complexed metal by the difference between the total concentration of dissolved and labile metal. Additionally, ligand complexation capacities (CC), such as the strength of the association of metals-ligands (logK'ML) and ligand concentrations (C L) were calculated via Ruzic's linearization method. Water samples were taken in winter and summer, and the results showed that for total and dissolved metals, Zn > Cu > Pb > Cd concentration. In general, higher concentrations of Cu and Zn remained complexed with the dissolved fraction, while Pb was mostly complexed with particulate materials. Chemical equilibrium modeling (MINEQL+) showed that Zn(2+) and Cd(2+) dominated the labile species, while Cu and Pb were complexed with carbonates. Zinc was a unique metal for which a direct relation between dissolved species with labile and complexed forms was obtained. The CC for ligands indicated a higher C L for Cu, followed by Pb, Zn, and Cd in decreasing amounts. Nevertheless, the strength of the association of all metals and their respective ligands was similar. Factor analysis with principal component analysis as the extraction procedure confirmed seasonal effects on water quality parameters and metal speciation. Total, dissolved, and complexed Cu and total, dissolved, complexed, and labile Pb species were all higher in winter, whereas in summer, Zn was mostly present in the complexed form. A high degree of deterioration of the reservoir was confirmed by the results of this study.

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The Paranaguá Estuarine Complex (PEC) includes the naturally oligotrophic (NO) Mel Island which is surrounded by sea grasses, a naturally eutrophic (NE) Benito Inlet adjacent to mangrove wetlands and the highly impacted eutrophic (IE) Paranaguá Bay, home of one of Brazil's largest ports. The results from this study indicate that reactive As and pyrite increase with sediment depth near Paranaguá port in the IE region. At the NE region, near a mangrove fringe, the reactive As, Fe, Mn and pyrite remained relatively high along the sediment column while near the sea grasses at NO the As contents were low. The degree of trace metal pyritization (DTMP) and the degree of pyritization (DOP) was highest at the IE site, slightly increasing with depth. These baseline results indicate that influence of trophic conditions and presence of marine vegetation may be directly related to As behavior in coastal systems.

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Fatty acids are the fundamental structural components of membrane lipids, and the degree of saturation of the long hydrocarbon chains in microalgae contributes to regulation of growth, biomass production and reproduction of aquatic consumers. This research aimed at evaluating the effects of cadmium (2×10(-8); 10(-7) mol L(-1) Cd) on lipid class and fatty acid composition of the microalga Chlorella vulgaris under varying phosphate (PO(4)(3-)) concentrations (6.0×10(-7) to 2.3×10(-4) mol L(-1)). Under PO(4)(3-) limitation and Cd stress, the storage lipid class triacylglycerol (TAG) was the most accumulated among the lipid classes. Fatty acid composition revealed that the degree of saturation increased with increasing Cd stress and PO(4)(3-) limitation. Decreasing PO(4)(3-) and increasing Cd concentrations resulted in higher saturated fatty acid (SAFA) and monounsaturated FA (MUFA) concentrations. Total polyunsaturated FA (PUFA) and ω3 PUFA, and PUFA:SAFA ratios were higher in the control (2.3×10(-4) mol L(-1) PO(4)(3-)) cells than in either PO(4)(3-) limitation or Cd stress, or in the combination of both stresses. Contrasting with all the other PUFAs, 18:2n - 6 increased as PO(4)(3-) limitation increased. A significant positive relationship of PUFAs, acetone mobile polar lipids (AMPL) and phospholipids (PL) with phosphate concentration in the culture media was obtained, while TAG concentrations had a positive association with total MUFA and SAFA. Total SAFA, 14:0, 18:1n - 9 and 18:2n - 6 were positively correlated with Cd and negatively with PO(4)(3-) concentrations. The microalga responded to combined PO(4)(3-) limitation and Cd exposure by increasing its total lipid production and significantly altering its lipid composition. The FA 18:2n - 6 may be considered a stress biomarker for PO(4)(3-) limitation and Cd stress in C. vulgaris.

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The acute toxicity of metals to Daphnia similis was determined and compared to other daphnid species to evaluate the suitability of this organism in ecotoxicology bioassays. To verify the performance D. similis in toxicity tests, we also investigated the effect of Pseudokirchneriella subcapitata at 1 x 10(5) and 1 x 10(6) cells ml(-1) on Cd and Cr acute toxicity to the cladoceran. Daphnid neonates were exposed to a range of chromium and cadmium concentrations in the absence and presence of the algal cells. Metal speciation calculations using MINEQL(+) showed that total dissolved metal concentrations in zooplankton culture corresponded to 96.2% free Cd and 100% free Cr concentrations. Initial total dissolved metal concentrations were used for 48 h-LC(50) determination. LC(50) for D. similis was 5.15 x 10(-7) mol l(-1) dissolved Cd without algal cells, whereas with 1 x 10(5) cells ml(-1), it was significantly higher (7.15 x 10(-7) mol l(-1) dissolved Cd). For Cr, the 48 h-LC(50) value of 9.17 x 10(-7) mol l(-1) obtained for the cladoceran in tests with 1 x 10(6) cells ml(-1) of P. subcapitata was also significantly higher than that obtained in tests without algal cells (5.28 x 10(-7) mol l(-1) dissolved Cr). The presence of algal cells reduced the toxicity of metals to D. similis, as observed in other studies that investigated the effects of food on metal toxicity to standard cladocerans. Comparing our results to those of literature, we observed that D. similis is as sensitive to metals as other standardized Daphnia species and may serve as a potential test species in ecotoxicological evaluations.

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The present study aimed to investigate the response of several life history parameters (body length and age of primipara, duration of embryonic development, maximum body length, reproduction and survival) of the zooplankton Ceriodaphnia silvestrii while exposed to copper contaminated algae Pseudokirchneriella subcapitata. In order to evaluate chronic exposure on the animal's life history, long-term experimental design was used. Cladocerans were fed with a dietary copper concentration ranging from 3 to 68 fg Cu cell(-1). Low waterborne copper exposure (around 10(-10)molL(-1) free Cu(2+) ions) was kept in the experiments. The results showed that by exposure of cladocerans during 7 days to contaminated food with 68 fg Cu cell(-1), a significant reduction in neonate production, survival and body size were obtained. Inhibition on egg production of zooplankton at 38 fg Cu cell(-1) were observed in 36 days chronic dietary copper exposure. The importance of entire life cycle study to better evaluate cladoceran responses to chronic dietary metal exposure was demonstrated.

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Changes in life cycle parameters (survival, growth, reproduction) and feeding rate of the tropical cladoceran Ceriodaphnia silvestrii as affected by Cu contaminated algae Pseudokirchneriella subcapitata were investigated. The dietary copper exposure ranged from 3 x 10(-15) to 68 x 10(-15) g Cu algal cell(-1). Low waterborne copper exposure (around 10(-10) mol l(-1) free Cu2+ ions) was kept in the experiments. The results show an increasing toxic effect on C. silvestrii with copper increase in algal cells; at the highest copper exposure, all life cycle parameters were significantly affected. A concentration of 38 x 10(-15) g Cu algal cell(-1) reduced egg hatching percentile and the number of neonates produced per female, but did not cause any statistically significant effect on animals survival nor to the number of eggs produced per female. The following sequence of events was observed from the lowest to the highest copper contamination: reproduction, feeding rate, body length and, at last, survival was affected. We conclude that algal cells are an important route of copper exposure and toxicity to cladocerans.

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