Physiological responses of Chlorella sorokiniana to copper nanoparticles.

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.

Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus.

Remote effects (occurring without physical contact) of two plant growth-promoting bacteria (PGPB) Azospirillum brasilense Cd and Bacilus pumilus ES4 on growth of the green microalga Chlorella sorokiniana UTEX 2714 were studied. The two PGPB remotely enhanced the growth of the microalga, up to six-fold, and its cell volume by about three-fold. In addition to phenotypic changes, both bacteria remotely induced increases in the amounts of total lipids, total carbohydrates, and chlorophyll a in the cells of the microalga, indicating an alteration of the microalga's physiology. The two bacteria produced large amounts of volatile compounds, including CO2, and the known plant growth-promoting volatile 2,3-butanediol and acetoin. Several other volatiles having biological functions in other organisms, as well as numerous volatile compounds with undefined biological roles, were detected. Together, these bacteria-derived volatiles can positively affect growth and metabolic parameters in green microalgae without physical attachment of the bacteria to the microalgae. This is a new paradigm on how PGPB promote growth of microalgae which may serve to improve performance of Chlorella spp. for biotechnological applications.

Nitrogen balancing and xylose addition enhances growth capacity and protein content in Chlorella minutissima cultures.

This study aimed to examine the metabolic changes in Chlorella minutissima cells grown under nitrogen-deficient conditions and with the addition of xylose. The cell density, maximum photochemical efficiency, and chlorophyll and lipid levels were measured. The expression of two photosynthetic proteins, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and the beta subunit (AtpB) of adenosine triphosphate synthase, were measured. Comparison of cells grown in medium with a 50% reduction in the nitrogen concentration versus the traditional medium solution revealed that the cells grown under nitrogen-deficient conditions exhibited an increased growth rate, higher maximum cell density (12.7×10(6)cellsmL(-1)), optimal PSII efficiency (0.69) and decreased lipid level (25.08%). This study has taken the first steps toward protein detection in Chlorella minutissima, and the results can be used to optimize the culturing of other microalgae.

Ecotoxicological characterization of a tropical soil after diazinon spraying.

The impact of diazinon spraying in an agricultural tropical soil through the evaluation of both the habitat and retention functions of the soil system was never reported. To fill this gap, five times the recommended dose of a commercial diazinon formulation was sprayed in an agricultural area of Costa Rica, and dilution gradients of the sprayed soil were prepared in the laboratory. Avoidance and reproduction tests with soil organisms (Eisenia andrei, Enchytraeus crypticus and Folsomia candida) to evaluate losses in terrestrial habitat function, and growth and reproduction tests with aquatic organisms (Chlorella vulgaris and Daphnia magna, respectively) to evaluate the retention function of soil were performed. Results demonstrated that regarding habitat function, F. candida reproduction was the most sensitive endpoint (EC(50) = 0.288 mg a.i./kg), followed by avoidance behaviour of E. andrei (EC(20) = 1.75 mg a.i./kg). F. candida avoidance and the reproduction of E. andrei and E. crypticus were not affected by diazinon. The toxicity tests with aquatic organisms showed that the soil retention function was insufficient to prevent effects of diazinon either on microalgae growth (EC(50) ≤ 0.742 mg/L and EC(20) ≤ 0.223 mg/L) and on the reproduction of the cladoceran (EC(50) ≤ 0.00771 mg/L and EC(20) ≤ 0.00646 mg/L). Results suggested that diazinon exerted toxic effects even at the dilution corresponding to the recommended dose, fact which makes its misuse an issue of environmental concern. Furthermore, the present study highlighted the importance and complementary nature of the assessment of both habitat and retention functions to an ecological risk assessment in tropical systems.

Oxidative stress induced by a commercial glyphosate formulation in a tolerant strain of Chlorella kessleri.

We studied the toxicity of a glyphosate formulation and provide evidence of metabolic alterations due to oxidative stress caused in a Chlorella kessleri tolerant strain by exposure to the herbicide. After 96 h of exposure to increasing concentrations of the herbicide (0-70 mg L(-1)) with alkylaryl polyglycol ether surfactant, growth was inhibited (EC50-96 h 55.62 mg L(-1)). Glyphosate increased protein and malondialdehyde content which was significantly higher than in the control at 50-70 mg L(-1). Superoxide dismutase and catalase activities and reduced glutathione levels increased in a concentration-dependant manner. Morphological studies showed increases in vacuolisation and in cell and sporangia sizes. The glyphosate formulation studied has a cytotoxic effect on C. kessleri through a mechanism that would involve the induction of oxidative stress. Upon glyphosate exposure, oxidative stress parameters such as SOD and CAT activities and MDA level could be more sensitive biomarkers than usually tested growth parameters in C. kessleri.

Oxidative stress and antioxidant defenses in two green microalgae exposed to copper.

The aim of this work was to assess the effects of 1 week copper exposure (6.2, 108, 210 and 414microM) on Scenedesmus vacuolatus and Chlorella kessleri. The strains showed different susceptibility to copper. Copper content was determined in both strains by total X-ray reflection fluorescence analysis (TXRF). In S. vacuolatus, the increase of medium copper concentration induced an augmentation of protein and MDA content, and a significant decrease in the chlorophyll a/chlorophyll b ratio. S. vacuolatus showed a significant increase of catalase activity in 210 and 414microM of copper, and a significant increment of SOD activity and GSH content only in 414microM of copper. On the contrary, C. kessleri did not show significant differences in these parameters between 6.2 and 108microM of copper. Increased copper in the environment evokes oxidative stress and an increase in the antioxidant defenses of S. vacuolatus.

In vivo microspectroscopy monitoring of chromium effects on the photosynthetic and photoreceptive apparatus of Eudorina unicocca and Chlorella kessleri.

In microorganisms and plants, chromium (Cr) is not essential for any metabolic process, and can ultimately prove highly deleterious. Due to its widespread industrial use, chromium has become a serious pollutant in diverse environmental settings. The presence of Cr leads to the selection of specific algal populations able to tolerate high levels of Cr compounds. The varying Cr-resistance mechanisms displayed by microorganisms include biosorption, diminished accumulation, precipitation, reduction of Cr(6+) to Cr(3+), and chromate efflux. In this paper we describe the effects of Cr(6+) (the most toxic species) on the photosynthetic and photoreceptive apparatus of two fresh water microalgae, Eudorina unicocca and Chlorella kessleri. We measured the effect of this heavy metal by means of in vivo absorption microspectroscopy of both the thylakoid compartments and the eyespot. The decomposition of the overall absorption spectra in pigment constituents indicates that Cr(6+) effects are very different in the two algae. In E. unicocca the metal induced a complete pheophinitization of the chlorophylls and a modification of the carotenoids present in the eyespot after only 120 h of exposition at a concentration equal or greater than 40 microM, which is the limit for total Cr discharge established by US EPA regulations. In C. kessleri, chromium concentrations a hundred times higher than this limit had no effect on the photosynthetic machinery. The different tolerance level of the two algae is suggested to be due to the different properties of the mucilaginous envelope and cell wall covering, respectively, the colonies of Eudorina and the single cells of Chlorella, which binds chromium cations to a different extent.

Chemical basis for the phytotoxicity of N-aryl hydroxamic acids and acetanilide analogues.

Germination inhibition activity of N-aryl hydroxamic acids and acetanilide analogues was measured on lettuce seeds (Lactuca sativa). Lipophilicity of the compounds was determined by HPLC. A correlation between lipophilicity values and percentage of germination inhibition was established. A model mechanism of action for auxin was used for analyzing the effect of the substituent at the alpha carbon atom (Ca) on the polarization of hydroxamic and amide functions in relation to the germination inhibition activity observed. Results suggest that the lipophilic and acidic properties play an important role in the phytotoxicity of the compounds. A test with the microalga Chlorella vulgaris was used to evaluate the potential herbicide activity of the hydroxamic acids and acetanilides.

Combined effects of algal (Chlorella vulgaris) density and ammonia concentration on the population dynamics of Ceriodaphnia dubia and Moina macrocopa (Cladocera).

Ammonia is a natural variable in ponds and lakes. Although an important source of nitrogen for microalgae, at high concentrations ammonia can affect the density and diversity of cladocerans. Using the cladocerans Ceriodaphnia dubia, and Moina macrocopa, the effect of nominal concentrations of ammonium chloride under acute and chronic exposures at different levels of algal food was tested. Regardless of food level, C. dubia was more sensitive than M. macrocopa to ammonia. In the absence of food, the median lethal concentration of ammonia (LC(50) 24 h) for C. dubia was (112 mg L(-1)) less than half that of M. macrocopa (232 mg L(-1)). When algal food (0.5 x 10(6) and 1.5 x 10(6) cells ml(-1) of Chlorella) was used, the LC(50) values were much higher for both cladoceran species. Based on the population growth studies, it was found that in controls of M. macrocopa an increase in the availability of Chlorella from 0.5 to 1.5 x 10(6) cells mL(-1) led to an increase in the maximum density from 4.7 +/- 0.2 to 16.4 +/- 1.2 ind.mL(-1), while in C. dubia the peak population density decreased from 7.9 +/- 0.6 to 5.0 +/- l.0 ind.mL(-1). An increase in ammonia concentration (10 to 40 mg L(-1) for C. dubia and 20 to 120 mg L(-1) for M. macrocopa) resulted in a corresponding decrease in peak population densities of the tested cladocerans. The rate of population increase (r) values for M. macrocopa in the controls ranged from 0.21 +/- 0.001 and 0.25 +/- 0.02 at 0.5 and 1.5 x 10(6) cells mL(-1) of Chlorella, respectively. The corresponding values of C. dubia in controls were 0.21 +/- 0.004 and 0.18 +/- 0.01. At 0.5 x 10(6) cells mL(-1) of algal food, the r values became negative under 40 and 120 mg L(-1) of ammonia for C. dubia and M. macrocopa, respectively. The role of algal food in ammonia toxicity to cladocerans was discussed. (c) 2002 Elsevier Science (USA).