Purified Pyropia yezoensis Pigment Extract-Based Tandem Dye Synthesis.

Red phycoerythrin (R-PE) is a highly valuable protein found in an edible seaweed, Pyropia yezoensis. It is used extensively in biotechnological applications due to its strong fluorescence and stability in diverse environments. However, the current methods for extracting and purifying R-PE are costly and unsustainable. The aim of the present study was to enhance the financial viability of the process by improving the extraction and purification of R-PE from dried P. yezoensis and to further enhance R-PE value by incorporating it into a tandem dye for molecular biology applications. A combination of ultrafiltration, ion exchange chromatography, and gel filtration yielded concentrated (1 mg·mL-1) R-PE at 99% purity. Using purified PE and Cyanine5 (Cy5), an organic tandem dye, phycoerythrin-Cy5 (PE-Cy5), was subsequently established. In comparison to a commercially available tandem dye, PE-Cy5 exhibited 202.3% stronger fluorescence, rendering it suitable for imaging and analyzes that require high sensitivity, enhanced signal-to-noise ratio, broad dynamic range, or shorter exposure times to minimize potential damage to samples. The techno-economic analysis confirmed the financial feasibility of the innovative technique for the extraction and purification of R-PE and PE-Cy5 production.

A rapid and multi-endpoint ecotoxicological test using Mychonastes afer for efficient screening of metals and herbicides.

Microalgal growth-based tests are international standards for ecotoxicity assessment; however, their long exposure times, large sample volumes, and reliance on a single growth-endpoint make them inadequate for rapid toxicity screening. Here, we aimed to develop a rapid and simple ecotoxicological test using the fast-growing green alga Mychonastes afer, with multiple endpoints-growth, lipid content, and photosynthesis. We exposed M. afer to two metals-silver and copper-and two herbicides-atrazine and diuron-for 24 h and identified the most sensitive and reliable endpoints for each toxicant: the maximum electron transport rate (ETRmax) for Ag, Cu and atrazine, and the lipid content for diuron. Lipid content was found to be both a sensitive and reliable biomarker, meeting the effluent limit guidelines in both the Republic of Korea and the USA. The sensitivity of M. afer to Ag and atrazine also closely matched the HC5 values derived from the species sensitivity distribution approach, confirming its reliability for setting regulatory concentrations of these contaminants. Our calculated predicted no-effect concentration (PNEC) values were similar to established European Union PNECs for Ag, Cu, atrazine, and diuron, underlining the utility of these biological endpoints for ecological risk assessment and regulatory decision making. This method required lower sample volume (2 mL vs 100 mL) and exposure time (24 h vs 72-120 h) than conventional green algal tests, and eliminated the need for labour-intensive cell counting, expensive equipment, and chlorophyll fluorescence measurement expertise. Overall, this M. afer test can be a valuable tool for the rapid screening of wastewater for metals and herbicides, contributing to environmental protection and management practices.

Solution-Based One-Step Preparation of Three-Dimensional Self-Assembled Octadecyl Silica Nanosquare Plate and Microlamella Structures for Superhydrophobic and Icephobic Surfaces.

Icephobic surfaces have gained immense attention owing to their significant roles in decreasing the energy consumption of refrigerators and in improving safety issues by preventing the formation of ice on them. Superhydrophobic surfaces incorporating micro- or nanoscale roughness and hydrophobic functional groups have been shown to prevent ice accumulation. Herein, we report a simple, low-cost, and solution-based one-step process for the production of superhydrophobic surfaces with three-dimensional (3D) self-assembled structures. The controlled hydrolysis and polycondensation of n-octadecyltrichlorosilane (OTS-Cl) in an acetone solution produced a highly uniform superhydrophobic surface on various substrates such as glass, metals, and polymers without the limitation of the surface curvature structure. The as-prepared 3D self-assembled surface exhibited a very high contact angle of 161.7° and a low contact hysteresis of 1.47°. The solvent type, H2O content in acetone, and carbon chain length of the silane compound were critical in the formation of self-assembled nanostructures. The thickness of the superhydrophobic 3D self-assembled structure could be varied by controlling the surface properties of the glass substrate. In addition, a novel octadecyl silica nanosquare plate structure was formed as an intermediate for the microlamella structure. The water drop impact experiments on the 3D self-assembled superhydrophobic glass substrates at low temperatures (T < -25 °C) showed that the as-prepared superhydrophobic glass possessed a high impalement threshold for water contact, resulting in excellent and stable icephobic properties. The preparation method proposed in this study is scalable and can be used on a flat glass surface or in a glass vial inside a glass tube. Moreover, it can be applied to various substrates such as metals and polyurethane surfaces with curvature. Therefore, the solution-based self-assembly method proposed in this study is a promising approach to produce superhydrophobic and icephobic surfaces on a wide range of substrates regardless of their structure and properties.

Rapid toxicity assessment of six antifouling booster biocides using a microplate-based chlorophyll fluorescence in Undaria pinnatifida gametophytes.

Biocides of antifouling agents can cause problems in marine ecosystems by damaging to non-target algal species. Aquatic bioassays are important means of assessing the quality of water containing mixtures of contaminants and of providing a safety standard for water management in an ecological context. In this study, a rapid, sensitive and inexpensive test method was developed using free-living male and female gametophytes of the brown macroalga Undaria pinnatifida. A conventional fluorometer was employed to evaluate the acute (48 h) toxic effects of six antifouling biocides: 4,5-Dichloro-2-octyl-isothiazolone (DCOIT), diuron, irgarol, medetomidine, tolylfluanid, zinc pyrithione (ZnPT). The decreasing toxicity in male and female gametophytes as estimated by EC50 (effective concentration at which 50% inhibition occurs) values was: diuron (0.037 and 0.128 mg l-1, respectively) > irgarol (0.096 and 0.172 mg l-1, respectively) > tolylfluanid (0.238 and 1.028 mg l-1, respectively) > DCOIT (1.015 and 0.890 mg l-1, respectively) > medetomidine (12.032 and 12.763 mg l-1, respectively). For ZnPT, 50% fluorescence inhibition of U. pinnatifida gametophytes occurred at concentrations above 0.4 mg l-1. The Undaria method is rapid, simple, practical, and cost-effective for the detection of photosynthesis-inhibiting biocides, thus making a useful tool for testing the toxicity of antifouling agents in marine environments.

Towards a multi-bioassay-based index for toxicity assessment of fluvial waters.

Despite their proven reliability for revealing 'acceptable' degrees of toxicity in waste- and reclaimed waters, bioassays are rarely used to assess the toxicity of hazardous contaminants present in natural waters. In this study, we used organisms from different trophic levels to assess the toxicity of water samples collected from four different South Korean rivers. The main objective was to develop a multi-descriptor index of toxicity for undiluted river water. The responses of six test organisms (Aliivibrio fischeri, Pseudokirchneriella subcapitata, Heterocypris incongruens, Moina macrocopa, Danio rerio and Lemna minor) after laboratory exposure to water samples were considered for this index, as well as the frequency of teratologies in diatom assemblages. Each individual test was attributed a toxicity class and score (three levels; no toxicity = 0, low toxicity = 1, confirmed toxicity = 2) based on the organism's response after exposure and a total score was calculated. The proposed index also considers the number of test organisms that received the highest toxicity score (value = 2). An overall toxicity category was then attributed to the water sample based on those two metrics: A = no toxicity, B = slight toxicity, C = moderate toxicity; D = toxicity and E = high toxicity. The susceptibility of the test organisms varied greatly and the sensitivity of their response also differed among bioassays. The combined responses of organisms from different trophic levels and with different life strategies provided multi-level diagnostic information about the intensity and the nature of contamination.

Testing the toxicity of metals, phenol, effluents, and receiving waters by root elongation in Lactuca sativa L.

Phytotoxicity tests using higher plants are among the most simple, sensitive, and cost-effective of the methods available for ecotoxicity testing. In the present study, a hydroponic-based phytotoxicity test using seeds of Lactuca sativa was used to evaluate the water quality of receiving waters and effluents near two industrial sites (Soyo and Daejon) in Korea with respect to the toxicity of 10 metals (As, Cd, Cr, Cu, Fe, Pb, Mn, Hg, Ni, Zn) and phenol, and of the receiving waters and effluents themselves. First, the L. sativa hydroponic bioassay was used to determine whether the receiving water or effluents were toxic; then, the responsible toxicant was identified. The results obtained with the L. sativa bioassay ranked the EC50 toxicities of the investigated metal ions and phenol as: Cd > Ni > Cu > Zn > Hg > phenol > As > Mn > Cr > Pb > Fe. We found that Zn was the toxicant principally responsible for toxicity in Daejeon effluents. The Daejeon field effluent had a higher Zn concentration than permitted by the effluent discharge criteria of the Ministry of Environment of Korea. Our conclusion on the importance of Zn toxicity was supported by the results of the L. sativa hydroponic assay, which showed that the concentration of Zn required to inhibit root elongation in L. sativa by 50% (EC50) was higher in the Daejeon field effluent than that of pure Zn. More importantly, we proved that the L. sativa hydroponic test method can be applied not only as an alternative tool for determining whether a given waste is acceptable for discharge into public water bodies, but also as an alternative method for measuring the safety of aquatic environments using EC20 values, with respect to the water pollutants investigated (i.e., Cd, Cr, Cu, Pb, Mn, Hg, Ni, Zn, and phenol).

The sensitivity of an hydroponic lettuce root elongation bioassay to metals, phenol and wastewaters.

The root elongation bioassay is one of the most straightforward test methods used for environmental monitoring in terms of simplicity, rapidity and economy since it merely requires filter paper, distilled water and Petri dishes. However, filter paper as a support material is known to be problematic as it can reduce the sensitivity of the test. The newly developed hydroponic method reported here differs from the conventional root elongation method (US EPA filter paper method) in that no support material is used and the exposure time is shorter (48 h in this test versus 120 h in the US EPA test). For metals, the hydroponic test method was 3.3 (for Hg) to 57 (for Cu) times more sensitive than the US EPA method with the rank orders of sensitivity, estimated from EC50 values, being Cu≥Cd>Ni≥Zn≥Hg for the former and Hg≥Cu≥Ni≥Cd≥Zn for the latter methods. For phenol, the results did not differ significantly; EC50 values were 124 mg L(-1) and 108-180 mg L(-1) for the hydroponic and filter paper methods, respectively. Lettuce was less sensitive than daphnids to wastewaters, but the root elongation response appears to be wastewater-specific and is especially sensitive for detecting the presence of fluorine. The new hydroponic test thus provides many practical advantages, especially in terms of cost and time-effectiveness requiring only a well plate, a small volume of distilled water and short exposure period; furthermore, no specialist expertise is required. The method is simpler than the conventional EPA technique in not using filter paper which can influence the sensitivity of the test. Additionally, plant seeds have a long shelf-life and require little or no maintenance.

Assessing industrial wastewater effluent toxicity using boosting algorithms in machine learning: A case study on ecotoxicity prediction and control strategy development.

Trace heavy metals have a tendency to persist in the effluent of industrial wastewater treatment facilities, leading to toxic effects on downstream water bodies. Traditional assessment methods relied on animal testing, but ethical concerns have rendered them unacceptable. An alternative solution is to evaluate wastewater toxicity using trophic-level aquatic organisms as bioassays. However, these bioassay methods involve costly and time-consuming chemical and biological analytical experiments. In this study, an artificial intelligence-powered water quality assessment (AiWA) approach is proposed for predicting industrial effluent ecotoxicity to further enhance the quick and cost-effective ecotoxicity assessment process. Initially, 99 samples were collected from industrial wastewater treatment plants representing 21 different industries in the Republic of Korea. Fourteen parameters were measured, encompassing both physicochemical and ecotoxicological aspects. Boosting algorithms, especially extreme gradient boosting (XGBoost) and adaptive boosting (AdaBoost), were employed for model development. XGBoost outperformed AdaBoost in terms of model performance. Feature selection analysis revealed that conductivity, copper, lead, selenium, pH, and zinc concentrations were the most suitable inputs for training the boosting model. The innovated XGBoost-based AiWA model demonstrated significantly higher performance (i.e., up to 80%) compared to conventional models with an R2 value of exceeding 0.94 and root mean square error of 3.5 toxicity unit for predicting the integrated toxicity unit (ITU). Additionally, pH and conductivity emerged as crucial indicators for reflecting ecotoxicity levels. Specially, this case study indicated that non-toxic/directly dischargeable levels (TU ≤ 1) were achieved when the pH ranged from 6.8 to 8.4 and the conductivity remained below 1651 µS/cm. These findings are expected to facilitate rapid and cost-effective detection of heavy metal ecotoxicity in industrial wastewater effluents, aiding decision-making in wastewater management.

Interactive Effects of Blue Light and Water Turbulence on the Growth of the Green Macroalga Ulva australis (Chlorophyta).

Macroalgal growth and yield are key to sustainable aquaculture. Although light and water turbulence are two important factors that affect algal productivity, research on their interaction is limited. Therefore, in this study, we investigated the effects of different wavelengths of light and the presence or absence of water turbulence on the growth of the green macroalga Ulva australis. Water turbulence was found to enhance the growth of U. australis irrespective of photosynthetic performance, but only in blue light cultures. The quantum dose of blue light required to induce 50% growth promotion was 1.02 mol m-2, which is comparable to the reported values for cryptochrome-mediated effects in other macroalgae. The combined effect of blue light and water turbulence led to the accumulation of photosynthesis-related proteins that support plastid differentiation and facilitate efficient photosynthesis and growth. Our findings thus highlight the potential of harnessing blue light and water turbulence to maximise macroalgal cultivation for sustainable and profitable algal aquaculture.

Rapid and Accurate Ecotoxicological Assessment of Heavy Metals Using Cyprinus carpio Cells.

Heavy metals have serious negative effects on various aquatic organisms, and therefore rapid and accurate ecotoxicological assessments of heavy metals are necessary. Fish-derived cells sensitive to heavy metals have been used as valuable tools for ecotoxicological assessments. However, this method requires a minimum toxicity treatment time of 96 h, which limits its use when rapid ecotoxicological assessments are required or ecotoxicological assessments of a large number of toxicants are performed. In this study, these limitations were overcome by adjusting parameters including the concentration of fetal bovine serum (FBS) in the medium and the treatment time of the toxicant. Specifically, we found that the maximum time for fish cells to remain unstarved was 6 h when using a medium containing 1% FBS. We applied both parameters to the ecotoxicological assessment (using a medium containing 1% FBS for the toxicity assessment and treating the toxicant for only 6 h). Surprisingly, these adjusted parameters allowed us to obtain faster and more accurate data than the traditional assessment. This improvement was due to the new assessment conditions that minimized the possibility that the growth-inducing effects of nutrients present in excess in the medium could interfere with the cellular response to the toxicant. The accuracy of this assessment was not limited to measuring the toxicity of heavy metals. In conclusion, we have established an ecotoxicity assessment that can generate rapid and accurate data on heavy metals. This new platform will become the cornerstone of rapid and accurate ecotoxicity assessments of heavy metals.

Metal Toxicity across Different Thallus Sections of the Green Macroalga, Ulva australis.

We aimed to identify functional differences between different sections of the thallus of Ulva australis and develop tissue-endpoint combinations to assess the toxicity of six metals (i.e., Ag, As, Cd, Cr, Cu, and Ni). EC50 values for these metals in three sections of the thallus of Ulva were obtained for multiple endpoints: relative growth rate (RGR), chlorophyll a fluorescence, pigment contents, and the expression of the photosynthesis-related gene, rbcL. The responses of the endpoints varied across the respective thallus sections; overall, the most toxic metals were Ag and Cu. These endpoints were the best for evaluating metal toxicity: ETRmax of the middle thallus sections for Ag toxicity; RGR of the middle thallus section for As and Cd; ETRmax of the marginal thallus section for Cr; Chl b contents of the marginal thallus section for Cu; RGR of the basal thallus section for Ni. The EC50 values for the inhibition of ETRmax in middle (0.06 mg∙L-1) and Chl b in the marginal thallus sections (0.06 mg∙L-1) were all lower than those of the quality standard for wastewater discharge values of Ag and Cu in Republic of Korea and the US, pointing to the suitability of U. australis-based endpoints for risk assessment.

Giant Duckweed (Spirodela polyrhiza) Root Growth as a Simple and Sensitive Indicator of Copper and Chromium Contamination.

Aquatic environment are often contaminated with heavy metals from various industrial sources. However, physicochemical techniques for pollutant detection are limited, thus prompting the need for additional bioassays. We investigated the use of greater duckweed (Spirodela polyrhiza) as a bioindicator of metal pollution. We exposed S. polyrhiza to four pollutants (namely, silver, cadmium, copper, and chromium) and assessed metal toxicity by measuring its frond area and the length of its regrown roots. The plant displayed significant differences in both frond size and root growth in response to the four metals. Silver was the most toxic (EC50 = 23 µg L-1) while copper the least (EC50 = 365-607 µg L-1). Direct comparisons of metal sensitivity and the reliability of the two endpoint assays showed that root growth was more sensitive (lower in terms of 50% effective concentration) to chromium, cadmium, and copper, and was more reliable (lower in terms of coefficient of variation) than those for frond area. Compared to conventional Lemna-based tests, the S. polyrhiza test is easier to perform (requiring only one 24-well plate, 3 mL of medium and a 72-h exposure). Moreover, it does not require livestock cultivation/maintenance, making it more suitable for repeated measurements. Measurements of S. polyrhiza root length may be suitable for assessment when copper and chromium in municipal and industrial wastewater exceed the environmentally permissible levels.

Qualitative and quantitative assessment of diatom deformities and protoplasmic condition under metal and metalloid stress.

Metals and metalloids are toxic, persistent, and non-biodegradable and can be biomagnified (e.g., Hg), and therefore pose a serious threat to the algal flora of aquatic ecosystems. This laboratory study tested the effects of metals (Zn, Fe, and Hg) and a metalloid (As) on the cell wall morphology and protoplasmic content of living cells of six widespread diatom genera over 28 days. Diatoms exposed to Zn and Fe had a higher frequency of deformed diatom frustules (> 1%) compared to the As, Hg, and control treatments (< 1%). Deformities in the valve outline and striae were found in all treatments, including the control, whereas deformed raphes and more than one type of deformity were more prevalent under Zn and Hg stress. The order of toxicity is as follows: Zn > Fe > Hg≈As. Deformities were more frequent in Achnanthes and Diploneis (adnate forms) than in the motile genera of Nitzschia and Navicula. The correlation between the % healthy diatoms and % deformities in all six genera showed a negative relationship with the integrity of protoplasmic content (i.e., greater alteration in protoplasmic content was associated with greater frustule deformation). We conclude that diatom deformities can be a good indicator of metal and metalloid stress in waterbodies and are very useful in the rapid biomonitoring of aquatic ecosystems.

Evaluating ecotoxicological assays for comprehensive risk assessment of toxic metals present in industrial wastewaters in the Republic of Korea.

Toxicity tests represent a rapid, user-friendly and cost-effective means to assess the impact of wastewater quality on aquatic ecosystems. There are not many cases where wastewater management standards are set based on various bio-based ecotoxicity values. Here, we tested a novel multitaxon approach to compare standard water quality indices to toxicity metrics obtained from ecotoxicity tests, conducted using aquatic organisms representing several trophic levels (Aliivibrio, Ulva, Daphnia, and Lemna), for 99 industrial wastewater samples from South Korea. For five wastewater samples, the concentrations of Se, Zn, or Ni exceeded the permissible limits (1, 5, and 3 mg L-1, respectively). All the four physiochemical water quality indices tested were positively correlated with Se and Pb concentrations. The toxicity unit (TU) scores indicated a declining sensitivity to pollutants, in the order Lemna (2.87) >Daphnia (2.24) >Aliivibrio (1.78) >Ulva (1.42). Significant correlations were observed between (1) Cd and Ni, and Aliivibrio, (2) Cu and Daphnia, (3) Cd, Cu, Zn, and Cr and Lemna, and (4) Cu, Zn, and Ni and Ulva. Daphnia-Lemna and Lemna-Ulva were found to be good indicators of ecologically harmful Se and Ni contents in wastewater, respectively. We suggest that regulatory thresholds based on these bioassays should be set at TU = 1 for all the species or at TU = 1 for Aliivibrio and Ulva and TU = 2 for Daphnia and Lemna, if the number of companies whose wastewater discharge exceeds the allowable TU levels is <1 % or 5 % of the total number of industries, respectively. Taken together, these findings could help in establishing a rapid, ecologically relevant wastewater quality assessment system that would be useful for developing strategies to protect aquatic ecosystems.

Molecular characterization of hot spring cyanobacteria and evaluation of their photoprotective compounds.

Phylogenetic analysis of 4 cyanobacterial strains isolated from hot springs in Rajgir, India, was carried out using the 16S rRNA gene (1400 bp). These strains were identified as members of Chroococcales ( Cyanothece sp. strain HKAR-1) and Nostocales ( Nostoc sp. strain HKAR-2, Scytonema sp. strain HKAR-3, and Rivularia sp. strain HKAR-4). Furthermore, we evaluated the presence of ultraviolet-screening and (or) photoprotective compounds, such as mycosporine-like amino acids (MAAs) and scytonemin, in these cyanobacteria by using high-performance liquid chromatography. Well-characterized MAAs, including the critical and highly polar compounds shinorine, porphyra-334, and mycosporine-glycine, as well as several unknown MAAs, were found in these hot-spring-inhabiting microorganisms. The presence of scytonemin was detected only in Scytonema sp. strain HKAR-3 and Rivularia sp. strain HKAR-4. The results indicate that hot spring cyanobacteria, namely Cyanothece, Nostoc, Scytonema, and Rivularia, belonging to different groups possess various photoprotective compounds to cope up with the negative impacts of damaging radiations.

Inter-population comparisons of copper resistance and accumulation in the red seaweed, Gracilariopsis longissima.

Copper (Cu) resistance and accumulation of five populations of the red seaweed Gracilariopsis longissima collected from sites in south west England (Fal Estuary, Helford Estuary and Chesil Fleet) that differ in their degree of Cu contamination was assessed under controlled laboratory conditions, on two separate occasions (April and October). The effects of a range of Cu concentrations (0-250 µg l(-1)) on relative growth rates was the same for all populations with reductions observable at concentrations as low as 12 µg l(-1) and cessation of growth at 250 µg l(-1). There was no significant difference in the calculated EC(50) values for the April and October samples, with means of 31.1 and 25.8 µg l(-1), respectively. Over the range of concentrations used in this study, copper content increased linearly and the pattern of accumulation was the same for all populations at both time periods. From the linear regressions of the pooled data a concentration factor of 2.25 × 10(3) was calculated. These results imply that G. longissima has an innate tolerance to Cu and that populations have not evolved copper-tolerant ecotypes. In laboratory studies, accumulated Cu was released when transferred to 'clean' seawater with approximately 80% being lost after 8 days, with no significant difference between populations in their response. The results from a 30 days in situ transplantation experiment using two populations from the Fal Estuary provided further evidence for dynamic changes in Cu content in response to changes in Cu bioavailability. The findings in this study are discussed in the context of implications for seaweed biomonitoring.

Commercial Potential of the Cyanobacterium Arthrospira maxima: Physiological and Biochemical Traits and the Purification of Phycocyanin.

Arthrospira maxima is a natural source of fine chemicals for multiple biotechnological applications. We determined the optimal environmental conditions for A. maxima by measuring its relative growth rate (RGR), pigment yield, and photosynthetic performance under different pH and temperature conditions. RGR was highest at pH 7-9 and 30 °C. Chlorophyll a, phycocyanin, maximal quantum yield (Fv/Fm), relative maximal electron transport rate (rETRmax), and effective quantum yield (ΦPSII) were highest at pH 7-8 and 25 °C. Interestingly, phycoerythrin and allophycocyanin content was highest at 15 °C, which may be the lowest optimum temperature reported for phycobiliprotein production in the Arthrospira species. A threestep purification of phycocyanin (PC) by ultrafiltration, ion-exchange chromatography, and gel filtration resulted in a 97.6% purity of PC.

A Ten-Minute Bioassay to Test Metal Toxicity with the Freshwater Flagellate Euglena agilis.

A chemical analysis of water quality cannot detect some toxicants due to time constraints, high costs, and limited interactions for detection. Bioassays would offer a complementary means to assess pollution levels in water. Euglena is a flagellate green alga and an excellent system for toxicity testing thanks to its ease of culture, rapid growth, and quick response to environmental stresses. Herein, we examined the sensitivity of E. agilis to seven heavy metals by analyzing six end-point parameters: motility, velocity, cell compactness, upward swimming, r-value, and alignment. Notably, the velocity of E. agilis was most sensitive to cadmium (96.28 mg·L-1), copper (6.51 mg·L-1), manganese (103.28 mg·L-1), lead (78.04 mg·L-1), and zinc (101.90 mg·L-1), while r-values were most sensitive to arsenic (12.84 mg·L-1) and mercury (4.26 mg·L-1). In this study, velocity and r-values are presented as useful biomarkers for the assessment of metal toxicity in Euglena. The metals As, Cd, Cu, and Pb were suitable for this test. The advantages of the ecotoxicity test are its rapidity: It takes 10 min to obtain results, as opposed to the typical 3-4 d of exposure time with intensive labor. Moreover, this test can be performed at room temperature under dark conditions.

Sensitivity of spore germination and germ tube elongation of Saccharina japonica to metal exposure.

The sensitivity of early life stages of the brown seaweed Saccharina japonica to six metals (Cd, Cu, Hg, Ni, Pb, Zn) and two waste-water samples were investigated and a new toxicity bioassay developed. The two endpoints used were spore germination and germ tube elongation with an exposure time of 24 h. Optimal test conditions determined for photon irradiance, pH, salinity and temperature were darkness, pH 8, 35‰ and 15°C, respectively. The toxicity ranking of five metals was: Hg (EC(50) of 41 and 42 µg l(-1)) > Cu (120 and 81 µg l(-1)) > Ni (2,009 and 1,360 µg l(-1)) > Zn (3,024 and 3,897 µg l(-1)) > Pb (4,760 and 4,429 µg l(-1)) > Cd (15,052 and 7,541 µg l(-1)) for germination and germ tube elongation, respectively. The sensitivities to Cd, Cu and Ni were greater in germ tube elongation than in germination process. When tested against two different waste-water samples (processed animal and printed circuit board waste-water) values of EC(50) were between 21.29 and 32.02% for germination and between 5.33 and 8.98% for germ tube elongation. Despite differences in their chemical composition, the toxic effects of waste-water samples, as indicated by EC(50) values, did not differ significantly for the same endpoints. The CV range for both germination and germ tube elongation was between 4.61 and 37.69%, indicating high levels of precision of the tests. The results compare favourably with those from more established test procedures employing micro- and macroalgae. The advantages and potential limitations of the bioassay for the assessment of anthropogenic impacts on coastal ecosystems and commercial cultivation areas in near-shore environments are discussed.

A comparative study on toxicity identification of industrial effluents using Daphnia magna.

In this study, acute toxicity monitoring and toxicity identification evaluation procedures were applied to identify causative toxicants in industrial effluents. Effluents from a metal plating factory and a rubber products factory were acutely toxic toward Daphnia magna and the toxicity varied over different sampling events (2.9-5.9 and 1.7-7.6 TU, respectively). For the rubber products effluent, it was confirmed that zinc (5.65-13.18 mg L(-1)) was found to be a major cause of toxicity, which is likely originated from zinc 2-mercaptobenzothiazole and zinc diethyldithiocarbamate used as vulcanization accelerators. For the metal plating effluent, it appeared that the presence of high concentrations of Cl(-) and SO(4)(2-) (8,539-11,400 and 3,588-4,850 mg L(-1), respectively) caused the observed toxicity. These toxicants likely originated from sodium bisulfate (NaHSO(3)) and sodium hypochlorite (NaOCl) used as reducing and oxidizing agents. Though copper was found to be present in levels much higher than the EC(50) (50% effective concentration) value, this was not attributable to the toxicity of metal plating effluent likely due to complexation with dissolved organic matter.