Marine Alga Dunaliella Tertiolecta Research Articles

Comparative Toxicity of Oil Spill Herding Agents to Aquatic Species.

Chemical herding agents are surfactant mixtures used to coalesce spilled oil and increase slick thickness to facilitate mechanical recovery or in situ burning. Only two herders are currently listed on the United States' National Oil and Hazardous Substances Pollution Contingency Plan or National Contingency Plan product schedule for potential use in spill response: the surface collecting agents Siltech OP-40™ and ThickSlick 6535™. Toxicity data for spill response agents are frequently available only for two estuarine species, mysid shrimp (Americamysis bahia) and inland silversides (Menidia beryllina), and are particularly limited for herding agents. Toxicity can vary over several orders of magnitude across product type and species, even within specific categories of spill response agents. Seven aquatic species were tested with both Siltech OP-40™ and ThickSlick 6535™ to evaluate acute herder toxicity and relative species sensitivity. The toxicity assessment included: acute tests with A. bahia and M. beryllina, the freshwater crustacean Ceriodaphina dubia, and the freshwater fish Pimephales promelas; development of the echinoderm Arbacia unctulate; and growth of a freshwater alga Raphidocelis subcapitata and marine alga Dunaliella tertiolecta. Siltech acute toxicity values ranged from 1.1 to 32.8 ppm. ThickSlick acute toxicity values ranged from 2.2 to 126.4 ppm. The results of present study show greater toxicity of Siltech compared to ThickSlick with estimated acute hazard concentrations intended to provide 95% species protection of 1.1 and 3.6 ppm, respectively, on empirical data and 0.64 and 3.3 ppm, respectively, with the addition of interspecies correlation data. The present study provides a greater understanding of species sensitivity of these two oil spill response agents. Environ Toxicol Chem 2022;41:1311-1318. © 2022 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Sensitivity of freshwater and marine green algae to three compounds of emerging concern

In this study, the toxicity of three compounds of emerging concern (CEC) belonging to different classes [metolachlor (herbicide), erythromycin (antibiotic) and triclosan (antiseptic)], were evaluated and compared using the freshwater alga Pseudokirchneriella subcapitata and the marine alga Dunaliella tertiolecta. Toxicity assays were performed by exposing algal cells, in exponential phase of growth, to the toxicants for 72 h (P. subcapitata) or 96 h (D. tertiolecta). The toxicant concentrations that induced an inhibition of 50% of algal growth (EC50) of P. subcapitata or D. tertiolecta were 118 and 11.3 × 103 μg L−1 for metolachlor (MTC), 38 and 5.75 × 103 μg L−1 for erythromycin (ERT) and 27.1 and 93 μg L−1 for triclosan (TCS), respectively. Based on these EC50 values, it was possible to hierarchize (decreasing order) the toxicity of the CEC studied: TCS > ERT > MTC. The EC50 values achieved for P. subcapitata were between 3.4- and 151-fold lower than those observed with D. tertiolecta, which demonstrated the higher sensitivity of the freshwater alga comparatively to the marine alga. All 72 h-EC10 or 72 h-EC50 values determined in this study with P. subcapitata are within the concentration range of these pollutants described in the literature, in ground and surface waters, which underlines the significance of this alga in the ecotoxicity assessment of freshwaters.

Colorimetric analysis is not sufficient to estimate bioavailable phosphorus in a hypersaline aquatic environment

Colorimetric analysis is commonly used to quantify P availability in aquatic environments. However, it may not be adequate to quantify the bioavailability of dissolved organic P compounds. This study aimed to investigate to what extent the colorimetrically measurable P fractions can be utilized by algae, with a particular emphasis on organic P compounds in a hypersaline aquatic environment. In this study, the growth of the marine alga Dunaliella tertiolecta Butcher was observed in axenic hypersaline cultures prepared with different organic P forms and related to the corresponding levels of colorimetrically measured P and total P. The malachite green method was used to colorimetrically analyze the different P forms and inductively coupled plasma-optical emission spectroscopy was used to quantify the total P. The results showed that only creatine P was colorimetrically detectable, of which 87% of its total concentration was measured. The growth of algae in the culture media with organic P compounds did not reflect the colorimetrically measurable organic P compounds. The results from this study imply that colorimetric analysis may not be sufficient to assess P availability to algae in hypersaline conditions where organic P components are the main source of P.

Quantitative structure–activity relationships (QSARs) using the novel marine algal toxicity data of phenols

The present study reports for the first time in its entirety the toxicity of 30 phenolic compounds to marine alga Dunaliella tertiolecta. Toxicity of polar narcotics and respiratory uncouplers was strongly correlated to hydrophobicity as described by the logarithm of the octanol/water partition coefficient (Log P). Compounds expected to act by more reactive mechanisms, particularly hydroquinones, were shown to have toxicity in excess of that predicted by Log P. A quality quantitative structure–activity relationship (QSAR) was obtained with Log P and a 2D autocorrelation descriptor weighted by atomic polarizability (MATS3p) only after the removal of hydroquinones from the data set. In an attempt to model the whole data set including hydroquinones, 3D descriptors were included in the modeling process and three quality QSARs were developed using multiple linear regression (MLR). One of the most significant results of the present study was the superior performance of the consensus MLR model, obtained by averaging the predictions from each individual linear model, which provided excellent prediction accuracy for the test set (Qtest2=0.94). The four-parameter Counter Propagation Artificial Neural Network (CP ANN) model, which was constructed using four out of six descriptors that appeared in the linear models, also provided an excellent external predictivity (Qtest2=0.93).The proposed algal QSARs were further tested in their predictivity using an external set comprising toxicity data of 44 chemicals on freshwater alga Pseudokirchneriella subcapitata. The two-parameter global model employing a 3D descriptor (Mor24m) and a charge-related descriptor (Cortho) not only had high external predictivity (Qext2=0.74), but it also had excellent external data set coverage (%97).

First toxicity data of chlorophenols on marine alga Dunaliella tertiolecta: Correlation of marine algal toxicity with hydrophobicity and interspecies toxicity relationships

The toxicity of phenol and 13 chlorinated phenols to the marine alga Dunaliella tertiolecta is presented for the first time. The newly generated marine algal toxicity data was found to correlate strongly with the widely used hydrophobicity parameter-the logarithm of the n-octanol-water partition coefficient (log K(OW)). Interspecies relationships using the new marine algal toxicity data of chlorophenols with the previously published data on bacterium (Vibrio fischeri), protozoan (Tetrahymena pyriformis), daphnid (Daphnia magna), freshwater alga (Pseudokirchneriella subcapitata), and fish (Pimephales promelas) revealed promising results that could be exploited in extrapolations using freshwater data to predict marine algal toxicity.

Naphthenic acid biodegradation by the unicellular alga Dunaliella tertiolecta

Naphthenic acids (NAs) are a major contributor to toxicity in tailings waste generated from bitumen production in the Athabasca Oil Sands region. While investigations have shown that bacteria can biodegrade NAs and reduce tailings toxicity, the potential of algae to biodegrade NAs and the biochemical mechanisms involved remain poorly understood. Here, we discovered that the marine alga Dunaliella tertiolecta is able to tolerate five model NAs (cyclohexanecarboxylic acid, cyclohexaneacetic acid, cyclohexanepropionic acid, cyclohexanebutyric acid and 1,2,3,4-tetrahydro-2-naphthoic acid) at 300 mg L −1, a level which exceeds that of any single or combination of NAs typically found in tailings ponds. Moreover, we show that D. tertiolecta can metabolize four of the model NAs. Analysis of NA-amended cultures of D. tertiolecta via low resolution gas chromatography–mass spectrometry allowed us to quantify decreasing NA levels, identify metabolites, and formulate putative mechanisms of biodegradation. Degradation of cyclohexanebutyric acid and cyclohexanepropionic acid proceeded via β-oxidation and resulted in the transient accumulation of cyclohexaneacetic acid and cyclohexanecarboxylic acid, respectively. Cyclohexanecarboxylic acid was metabolized via 1-cyclohexenecarboxylic acid suggesting that further degradation may occur by step-wise β-oxidation. When D. tertiolecta was inoculated in the presence of oil sands tailings water from the Athabasca region, biodegradation of single-ring NAs was observed relative to controls. This result corroborates the trend we observed with the single-ring model NAs.

Characterisation of a bioassay using the marine alga Dunaliella tertiolecta associated with spectroscopic (visible and infrared) detection

This paper describes an ecotoxicological test using the alga Dunaliella tertiolecta associated with visible (VIS) and infrared (FTIR) spectroscopic detection. The estimation of toxicity is performed by measuring the cell density at 432 nm instead of 665 nm the results are comparable with those obtained by conventional visual or automatic estimation of celle density. FTIR spectroscopy gives an insight to molecular modifications in cell of Dunaliella tertiolecta caused by pollutants. A relevant result obtained by FTIR is the high similarity observed between the molecular modifications caused by toxic effect and the molecular modifications caused by samples with a stimulation (i.e., hormetic) effect.

Exposure of Dunaliella tertiolecta to Lead and Aluminum: Toxicity and Effects on Ultrastructure

The growth response of the marine alga Dunaliella tertiolecta to different concentrations of lead and aluminum was investigated. Both metals had a stimulatory effect at low concentration and an inhibitory effect at high concentration (hormesis). The IC25 values of lead are 8.43, 7.29, and 6.74 mg L-1 for 24, 48, and 72 h, respectively. The corresponding values for aluminum are 30.54, 22.42, and 18.16 mg L-1. Although it seems that the two metals are not directly toxic to the alga at the concentrations found in the environment, as implied by the IC25 values and the environmental concentrations of the metals, low concentrations of both metals, alone and in combination, affected the ultrastructure. The growth of batch-grown cells exposed to 0.5 mg L-1 lead and aluminum, alone and combined, during the 24-h exponential phase was investigated. The same cells were also examined under an electron microscope to determine the biological effects of the two metals on the ultrastructure. The most obvious effects of lead were disrupted thylakoidal membranes, accumulated polyphosphate bodies and vacuoles, and lead precipitates on the cell surface. These ultrastructural alterations were partially present in aluminum-treated and lead-aluminum-treated cells. In joint exposure, the most important change was the lysis of the cell membrane. Aluminum and lead seem to act synergistically on the cell membrane leading to cell membrane lysis.

A case study on algal response to raw and treated effluents from an aluminum plating plant and a pharmaceutical plant

The algal growth responses to the effluents of an aluminum plating plant and to the wastewater from an analgesic/antiinflammatory-drug-producing pharmaceutical plant were investigated. Growth response of the marine alga Dunaliella tertiolecta was monitored by measuring the two response parameters optical density (OD 640) and in vitro chlorophyll fluorescence for a period of 14 days. Generally, the two response measurements gave similar results for all effluents but the raw effluents of the aluminum plating plant due to the composition of the wastewater. All wastes affected algal growth either by inhibition only or by stimulation at low concentrations and inhibition at high concentrations. Since pollutant tolerance of algae biased toxicity test results, acclimation of algae to the raw effluent of the aluminum plating plant was examined. Although the water quality parameters of treated effluent of both plants were in the permitted range reported by the Turkish Water Pollution Control Act, they inhibited growth at higher concentrations, implying that the two treatment plants were inefficient. Therefore, the importance of toxicity tests in wastewater discharge regulations was emphasized.

Influence of Chemical and Physical Factors on Long-Term Delayed Fluorescence in Dunaliella tertiolecta

The long-term delayed fluorescence in the marine alga Dunaliella tertiolecta (Chlorophyta) was investigated for the influence of herbicide DCMU, reference toxicant 3,5-DCP, temperature, excitation light spectrum, and intensity. We observed a strong transient peak in otherwise monotonic decay kinetics, which was almost insensitive to illumination light spectra as opposed to Prorocentrum minimum, where the peak was induced only by far-red illumination. DCMU and 3,5-DCP influenced the DF kinetics of D. tertiolecta in a dose-dependent and qualitatively different way. We observed strong influence of temperature on the transient peak position (Q10 = 2.6, Ea = 71.5 kJ/mol, which are in the expected range of plastoquinone-PSII reactions). At the lowest illumination intensity, the peak was less pronounced, whereas the decay kinetics of the DCMU-treated samples was unchanged. The approach and results presented in this study could be used in modeling, especially where systematic variations in external influences present a decision-point between different models.

Removing UV‐A and UV‐C Radiation from UV‐B Fluorescent Lamp Emissions. Differences in the Inhibition of Photosynthesis in the Marine Alga Dunaliella tertiolecta Using Chromate Versus Cellulose Acetate‐Polyester Filters¶

ABSTRACTUltraviolet‐B (UV‐B; 280‐320 nm)‐emitting lamps unavoidably emit ultraviolet‐A (UV‐A; 320‐400 nm) and ultraviolet‐C (UV‐C; < 280 nm) radiation. Short‐wavelength‐blocking filters are generally used to limit the wave bands of UV under investigation. The widespread use of such filters means that all exposures to UV‐B radiation will have a significant UV‐A component. Therefore, the physiological effects unique to UV‐B exposure are difficult to clearly isolate. This study presents a method to remove the UV‐A and UV‐C “contamination” using a liquid potassium chromate (K2CrO4) filter, thus allowing more direct assessment of the effects of UV‐B exposure. Cultures of the green marine alga Dunaliella tertiolecta were grown in the absence of UV radiation. Sunlamps supplied the UV radiation for a 24 h exposure (solar radiation was not used in this study). The UV radiation was filtered either by the standard method (i.e. cellulose acetate (CA) with polyester = Mylar controls) or by a liquid filter of potassium chromate. Photosynthetic responses were compared. Major decreases in the ratio of variable to maximal fluorescence in dark‐adapted cells and photosynthetic capacity were observed in CA‐filtered cultures, whereas no change was observed in cells exposed to the same UV‐B flux with the UV‐A removed by K2CrO4. The use of a CA filter with a Mylar control does not link results unequivocally to UV‐B radiation. Such results should be interpreted with caution.

Removing UV-A and UV-C Radiation from UV-B Fluorescent Lamp Emissions. Differences in the Inhibition of Photosynthesis in the Marine Alga Dunaliella tertiolecta Using Chromate Versus Cellulose Acetate–Polyester Filters

Ultraviolet-B (UV-B; 280–320 nm)–emitting lamps unavoidably emit ultraviolet-A (UV-A; 320–400 nm) and ultraviolet-C (UV-C; <280 nm) radiation. Short-wavelength–blocking filters are generally used to limit the wave bands of UV under investigation. The widespread use of such filters means that all exposures to UV-B radiation will have a significant UV-A component. Therefore, the physiological effects unique to UV-B exposure are difficult to clearly isolate. This study presents a method to remove the UV-A and UV-C “contamination” using a liquid potassium chromate (K2CrO4) filter, thus allowing more direct assessment of the effects of UV-B exposure. Cultures of the green marine alga Dunaliella tertiolecta were grown in the absence of UV radiation. Sunlamps supplied the UV radiation for a 24 h exposure (solar radiation was not used in this study). The UV radiation was filtered either by the standard method (i.e. cellulose acetate (CA) with polyester = Mylar controls) or by a liquid filter of potassium chromate. Photosynthetic responses were compared. Major decreases in the ratio of variable to maximal fluorescence in dark-adapted cells and photosynthetic capacity were observed in CA-filtered cultures, whereas no change was observed in cells exposed to the same UV-B flux with the UV-A removed by K2CrO4. The use of a CA filter with a Mylar control does not link results unequivocally to UV-B radiation. Such results should be interpreted with caution.

Removing UV-A and UV-C radiation from UV-B fluorescent lamp emissions. Differences in the inhibition of photosynthesis in the marine alga Dunaliella tertiolecta using chromate versus cellulose acetate-polyester filters.

Ultraviolet-B (UV-B; 280-320 nm)-emitting lamps unavoidably emit ultraviolet-A (UV-A; 320-400 nm) and ultraviolet-C (UV-C; <280 nm) radiation. Short-wavelength-blocking filters are generally used to limit the wave bands of UV under investigation. The widespread use of such filters means that all exposures to UV-B radiation will have a significant UV-A component. Therefore, the physiological effects unique to UV-B exposure are difficult to clearly isolate. This study presents a method to remove the UV-A and UV-C "contamination" using a liquid potassium chromate (K(2)CrO(4)) filter, thus allowing more direct assessment of the effects of UV-B exposure. Cultures of the green marine alga Dunaliella tertiolecta were grown in the absence of UV radiation. Sunlamps supplied the UV radiation for a 24 h exposure (solar radiation was not used in this study). The UV radiation was filtered either by the standard method (i.e. cellulose acetate (CA) with polyester = Mylar controls) or by a liquid filter of potassium chromate. Photosynthetic responses were compared. Major decreases in the ratio of variable to maximal fluorescence in dark-adapted cells and photosynthetic capacity were observed in CA-filtered cultures, whereas no change was observed in cells exposed to the same UV-B flux with the UV-A removed by K(2)CrO(4). The use of a CA filter with a Mylar control does not link results unequivocally to UV-B radiation. Such results should be interpreted with caution.

Long-term hyposaline and hypersaline stresses produce distinct antioxidant responses in the marine alga Dunaliella tertiolecta

Tolerance to salinity stress in higher plants correlates to levels of antioxidant enzymes and/or substrates. Do hyperosmotic and hypoosmotic stress induce antioxidant responses in salt tolerant algae, and if so, are these responses the same for both excess and minimal salinity? To answer these questions, cultures of the marine alga Dunaliella tertiolecta (Chlorophyta) were grown in seven salinities covering a 60-fold range from 0.05 to 3.0 mol/L NaCl. Long-term effects of salinity on growth and antioxidant parameters were determined. Growth rates were reduced at the salinity extremes (0.05 mol/L NaCl and 3 mol/L NaCl) indicating the cultures were stressed. The levels of six antioxidant enzymes and three antioxidant substrates were quantified at these growth salinities. Compared to growth at optimum salinities (i.e. 0.2-0.5 mol/L NaCl), high salinities produced a 260% increase in monodehydroascorbate reductase, a doubling of ascorbate peroxidase activity and a three-fold increase in the rate of dark respiration. Cells acclimated to low growth salinities (hyposaline stress, i.e. < 0.2 mol/L NaCl) showed major increases in glutathione and alpha-tocopherol coupled with decreases in Fv/Fm ratios and in total and reduced ascorbate compared to moderate and high external salinities. Cell volumes remained unchanged, except at the lowest salinity where they doubled. Catalase, superoxide dismutase, dehydroascorbate reductase and glutathione reductase activities were not altered by extreme salinities. The involvement of oxidative stress at both salinity extremes is implied by the alterations in antioxidant enzymes and substrates, but the specific changes are very different between hypo and hypersaline stresses.

Probing cell surface charge by scanning electrode potential.

A static mercury electrode was used for measurement of double-layer charge displacement signals caused by living plant cells of the unicellular marine alga Dunaliella tertiolecta. By scanning the electrode potential a point is reached where the charge density of a plant cell compensates the electrode charge density. The experimentally determined values of surface charges for unicellular marine alga Dunaliella tertiolecta cells are -0.63 and -0.75 nuC/cm(2) in 0.1 M NaCl and 1 M NaCl solutions, respectively.

Laboratory bioaccumulation of copper, lead and selenium in the marine alga dunaliella tertiolecta metal pair situation

Individual and combined effects on the bioaccumulation of three heavy metals; namely, copper, lead and selenium, in the unicellular green alga Dunaliella tertiolecta were determined in natural sea water enriched with f/ 2 medium. Exponentially growing cells of alga were exposed to the three metals in the form of Cu(NO3)2 5H2O, Pb(NO3)2 and Na2SeO3 5H2O for 24 h. The pH effect on the bioaccumulation of each metal has been investigated. Total copper and lead concentrations were measured by atomic absorption spectrophotometer, whereas total selenium concentration was measured by spectrofluorometer. The bioaccumulation of metals occurred in a dose‐dependent manner. When pH decreased, the bioaccumulation of copper decreased significantly, but the bioaccumulation of lead and selenium almost stayed constant. The presence of copper interacted antagonistically with lead in various bimetallic combinations. Lead showed no effect on the bioaccumulation of copper. Coexistence of selenium with lead and copper showed antagonistic behaviour on the growth of alga microscopically.

Low-CO2-inducible protein synthesis in the green alga Dunaliella tertiolecta

In the green marine alga Dunaliella tertiolecta, a CO2-concentrating mechanism is induced when the cells are grown under low-CO2 conditions (0.03% CO2). To identify proteins induced under low-CO2 conditions the cells were labelled with 35SO4 2−, and seven polypeptides with molecular weights of 45, 47, 49, 55, 60, 68 and 100 kDa were detected. The induction of these polypeptides was observed when cells grown in high CO2 (5% CO2 in air) were switched to low CO2, but only while the cultures were growing in light. Immunoblot analysis of total cell protein against pea chloroplastic carbonic anhydrase polyclonal antibodies showed immunoreactive 30-kDa bands in both high- and low-CO2-grown cells and an aditional 49-kDa band exclusively in low-CO2-grown cells. The 30-kDa protein was shown to be located in the chloroplast. Western blot analysis of the plasmamembrane fraction against corn plasma-membrane AT-Pase polyclonal antibodies showed 60-kDa bands in both high- and low-CO2 cell types as well as an immunoreactive 100-kDa band occurring only in low-CO2-grown cells. These results suggest that there are two distinct forms of both carbonic anhydrase and plasma-membrane ATPase, and that one form of each of them can be regulated by the CO2 concentration.

Chemoreception in the green algaDunaliella tertiolecta

A capillary assay was employed to quantify positive chemotactic responses in the motile, unicellular, marine algaDunaliella tertiolecta. Among a wide range of inorganic and organic compounds tested, only ammonium ion,l-tyrosine,l-tryptophan, andl-phenylalanine were found to be major atractants for the chlorophyte.l-Methionine andl-cysteine weakly attracted the alga at 10−3 M. The minimum concentration of the major attractants needed to elicit an observable chemotactic response was approximately 10−6 M. The maximum response occurred when the capillaries contained 10−5 Ml-tyrosine orl-tryptophan, 10−4 Ml-phenylalanine, and 10−3 M ammonium chloride. The other amino acids, carbohydrates, B-vitamins, urea, and nitrate were among the chemicals that failed to attractD. tertiolecta. The alga apparently possesses one chemoreceptor that binds ammonium ion only, and another chemoreceptor that binds the three aromatic amino acids.

Predicting Production in Light-Limited Continuous Cultures of Algae

Equations relating productivity, growth rate, cell concentration, and light absorption lead to the prediction that, when incident light is below saturating intensity, maximal productivity will occur at half the maximal growth rate. The freshwater alga Chlorella pyrenoidosa TX71105 and the marine alga Dunaliella tertiolecta were grown in a small continuous culture apparatus with turbidostatic control. With both cultures, the cell concentration showed a linear decrease with dilution rate. Productivity was maximal at about one-half the maximal dilution rate. Average mass per cell increased near the maximal dilution rate, causing some asymmetry in the productivity versus dilution rate curve. The chlorophyll content per unit mass decreased in this region, but the chlorophyll content per cell remained constant. Best production rate in a light-limited algal culture was obtained when the growth rate at very low cell concentration was determined in the apparatus and the dilution rate was set at one-half that value.