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.

Potential of cyanobacterial biofilms in phosphate removal and biomass production.

Four cyanobacterial biofilms, raised from cyanobacterial mats and dominated by Phormidium and Oscillatoria spp., were successfully grown attached to polyester mesh discs, and were tested for their probable application in [Formula: see text] -P removal from domestic sewage and other nutrient enriched wastewaters. Biofilm # 2, dominated by Phormidium fragile, best removed [Formula: see text] -P; nevertheless, some of it also grew outside the substrate making harvesting difficult. Other biofilms also efficiently removed [Formula: see text] -P from the medium in the following order: Biofilm # 1 > Biofilm # 3 > Biofilm # 4. Their growths were restricted to discs and are therefore better candidates as they can be efficiently harvested after [Formula: see text] -P removal. [Formula: see text] -P removal was primarily due to its uptake during growth of the biofilm rather than because of precipitation as pH of the medium remained <8.5. [Formula: see text] -N concentration in the medium determined [Formula: see text] -P removal efficiency of the test biofilms and therefore optimum N:P ratio is necessary for optimizing [Formula: see text] -P removal. The test biofilms could also efficiently remove [Formula: see text] -N from the medium.

Response of a phytoplanktonic assemblage to copper and zinc enrichment in microcosm.

The response of a laboratory-raised phytoplankton assemblage to copper and zinc enrichment was studied. Higher intracellular accumulation of both the test metals caused disappearance of metal sensitive species, loss of diversity and species richness, reduced growth rate, Chl a and biovolume; however, the community could recover after 14 days of incubation. Cyanobacteria showed marked sensitivity to both the test metals besides some diatoms, such as, Cyclotella meneghiniana and Melosira granulata. Metal enrichment enhanced the relative abundance of species like Scenedesmus quadricauda, Oocystis borgei, Achnanthes exigua, Fragilaria capucina and Nitzschia amphibia, and these were apparently metal tolerant. Cu and Zn stress induces formation of lipid bodies (bigger in size as well as in number) and morphological abnormalities in diatoms. Among these two metals, Cu impact was higher than Zn despite the fact that the intracellular accumulation of Zn was higher than Cu. Deformed raphe and mixed deformities in diatoms were exclusively found under heavy metal stress which was well supported by regression analysis. Finally the present study gives new insight for using diatoms as an effective tool for biomonitoring and biofuel production.

Diatoms as indicators of environmental health on Korean islands.

Diatoms are highly sensitive to perturbations in their environment and are thus useful as bioindicators for anthropogenic impacts such as pollution. However, there is no consensus about what aspects of diatom populations to measure (e.g., diversity, physiology, or morphology) and efficient and reliable survey protocols are lacking. Here, we evaluated the ecological status of diatom communities using both traditional and relatively novel methods on two islands (Deokjeok island and Daeijak island) affected by anthropogenic activities due to extensive agricultural practices and exploitation and that are under consideration for representative touristic sites in South Korea. Dissolved concentrations of metals and metalloid (As, Cu, Cr, Cd, Ni, Hg, Pb, and Zn) were below the ecological screening and toxicity reference values in water fractions but were above these values for sediment, particularly at one island, Deokjeok. The tested methods were generally consistent in finding little evidence for disruption of diatom communities, with dominance by Navicula and Gyrosigma, relatively high diversity, and typical abundance of lipid bodies and morphological deformities. However, analysis of lipid bodies and morphological deformities suggested greater potential anthropogenic disturbance at one site in Deokjeok. Future planning is required to ensure the maintenance of the near-pristine environments present on these islands.

Assessment of five live-cell characteristics in periphytic diatoms as a measure of copper stress.

Metal pollution of fluvial systems remains a major problem and biomonitoring can be a useful tool for assessing the metal contamination. To assess their potential as new bioindicators of copper stress, we treated a field-collected live periphytic diatom community (dominated by Amphora, Navicula, and Nitzschia) with dissolved Cu under optimal growth conditions. We studied the effects of Cu on five live-cell attributes: motility, protoplasmic content, lipid body number and biovolume, and frustule morphology. In all three genera, motility and protoplasmic content decreased, whereas the LB number, biovolume and deformity increased when Cu and exposure time increased. The sensitivity to Cu was highest for % MF, % CPC and % BCLB in Navicula and the LB number and deformity in Nitzschia. Amphora appeared to be more tolerant to Cu in comparison with other genera. The five cell attributes were inter-related. A heatmap showed that a recommended indicator for rapid screening of Cu toxicity was % BCLB for Amphora and % MF for Navicula and Nitzschia. % MF might be the most common representative indicator that can be applied to all three genera to evaluate the lethal effects of Cu stress if only one of the five cell attributes must be selected.

Assessment of metal contamination in water and sediments from major rivers in South Korea from 2008 to 2015.

This study is the first report to evaluate (8 years data) the contamination degree and distribution characteristics of metals in the surface water and sediments of four Korean rivers (Nakdong, Yeongsan, Geum, and Han). Eight years of data were evaluated, and metal concentrations in the river water were found to be below permissible limits but high enough to cause detrimental effects (under chronic exposure) to aquatic organisms. The analysis of metals in the river sediments showed the following trend: Zn > Cu > Cd > Pb > Ni > As > Cr > Hg. The concentrations of metals in sediments (especially in the Geum and Han rivers) were above the permissible limits reported by international agencies. Concentrations of Cu, Ni, and Zn were high enough to pose risks to aquatic communities. In sediments, metals pollution was also evaluated using different indices, such as enrichment factor (EF), geoaccumulation index (Igeo), contamination factor (CF), degree of contamination (Cd), modified degree of contamination (mCd), and pollution load index (PLI). The CF, EF, and Igeo indices demonstrated that most of the river sediment samples were moderately to heavily contaminated by Cd, Cu, Pb, and Zn. The PLI values were above one in the Geum and Han river sediments, which indicated polluted conditions. Similarly, Cd indicated a considerable to very high degree of contamination, while mCd indicated a low to moderate degree of contamination in all four river sediments. Finally, it was found that the extent of metals pollution in the Korean rivers reached a critical condition, which could be detrimental to the biota of the rivers, as well as to humans in the long term.

Reappraisal of the toxicity test method using the green alga Ulva pertusa Kjellman (Chlorophyta).

This study was aimed to develop an objective way of quantifying the reproductive status of the green macroalga, Ulva pertusa using a vital stain and programmed automated analysis (by Image J program). The EC50 values (with 95% CI), the concentrations of toxicants inducing a reduction of 50% in sporulation after 96 h exposure, from the newly developed method were similar to those obtained by the conventional method: 0.651 (0.598-0.705) mg l-1 for Cd, 0.144 (0.110-0.162) mg l-1 for Cu, 0.180 (0.165-0.195) mg l-1 for atrazine, 0.076 (0.049-0.094) mg l-1 for diuron and 30.6 (26.5-34.4) ml l-1 for DMSO, respectively. When the EC50 values from this study were compared to that those from literatures, the sensitivity for some toxicants was similar or higher than that of U. fasciata (1.930 mg l-1 for germination for Cd), U. armoricana (0.250 mg l-1 for Fv/Fm for Cu), U. reticulata (0.126-1.585 mg l-1 for growth for Cu), and U. intestinalis (0.650 mg l-1 for Fv/Fm for atrazine). The subjective views of the experimental performers can be eliminated using the newly developed method. The Ulva method gave consistent responses to Cu and Cd of internationally allowable ranges for effluents, implying that the method is a useful tool for monitoring industrial wastewaters containing these metals.

Metal toxicity and recovery response of riverine periphytic algae.

In the present study, in situ assessment of metal (Cu and Zn) toxicity followed by their recovery response was examined in periphyton dominated by diatoms. For doing so, metal diffusing substrates (MDS) were constructed and deployed in the river water for 6 weeks (3 weeks stress and 3 weeks recovery after replacing metal solution from the MDS). The use of MDS ensured that colonised periphyton on metal diffusing and control substrates were exposed to similar environmental conditions. The metal toxicity and recovery response of the community was examined in terms of traditional algal community parameters (biovolume, species richness, Shannon index, relative abundance) as well as with the newer non-taxonomical parameters (deformities and lipid bodies in diatoms). Both traditional and non-taxonomical parameters indicated complete recovery (from metal toxicity) of periphytic communities after 3 weeks following the withdrawal of Cu and Zn solution from the diffusing substrates. Newer non-taxonomical parameters, such as, deformities and lipid bodies, provide a new insight to understand metal toxicity and recovery response of diatom assemblages (the dominant autotrophs in the periphyton community) because these features are directly visible in live frustules, need no expertise in identification of diatoms and can be globally assessed with simple protocol. The experimental loss of metal pollutants and the constant immigration of algae (not previously exposed to high levels of metals) in fluvial systems aided periphyton recovery. Lastly, it is found that periphytic biofilms (dominated by diatoms) proved to be good bioindicators of metal toxicity and recovery in fluvial ecosystem.

Evaluating features of periphytic diatom communities as biomonitoring tools in fresh, brackish and marine waters.

The aims of this study were to assess the biodiversity of periphytic diatom assemblages in fresh, brackish and marine waterbodies of Korea, and to assess the effect of environmental and anthropogenic factors on parameters such as the quantity and biovolume of lipid bodies and deformations of diatoms as early warning measures of anthropogenic impact. Diatom samples were collected from 31 sites (14 freshwater, 10 brackish and 7 marine), which included less impacted (upstream) and impacted (downstream) sites in each water type. Our results showed higher abundance and biodiversity of periphytic diatoms at the less impacted sites in terms of species richness, Shannon index, cell count and biovolume of the communities than at the impacted sites for freshwater and estuarine sites, but not for marine sites. 84 diatom species were noted in freshwater, 80 in brackish water and 40 in marine waters. In comparison to diatoms of the impacted sites, those of less impacted freshwater, brackish and marine sites had less lipid bodies (also less biovolume) and a lower percentage of teratological frustules, and showed more mobile forms in the community. Principal component analysis (PCA) also showed clear segregation of impacted from less impacted sites by the extent of the presence of lipid bodies (higher both in number and biovolume) and deformities in diatom frustules. Pearson correlation analysis revealed that lipid body induction and deformities were positively correlated with metals (Cd, Co, Cr, Cu, Fe, Pb and Zn) and nutrients (total phosphorus and total nitrogen), whereas they showed negative correlation with salinity, dissolved oxygen, suspended solutes and pH. Life-forms, lipid bodies and deformities in diatoms may be an effective biomonitoring tool for assessing biological effects of pollutants in non-marine aquatic ecosystems in Korea.

The use of diatoms in ecotoxicology and bioassessment: Insights, advances and challenges.

Diatoms are regularly used for bioassessment and ecotoxicological studies in relation to environmental and anthropogenic disturbances. Traditional taxonomical diatom parameters (cell counts, biovolume estimates, species richness, diversity indices and metrics using sensitive and tolerant diatom species) are regularly used for these studies. In the same context, very less focus was given on new endpoints of diatoms (life-forms, nuclear anomalies, alteration in photosynthetic apparatus shape, motility, lipid bodies, size reduction and deformities), in spite of their numerous merits, such as, their easiness, quickness, cheapness, global acceptation and no especial training in diatom taxonomy. In this review we analyzed 202 articles (from lab and field studies), with the aim to investigate the bioassessment and ecotoxicological advancement taken place in diatom research especially in terms of exploring new endpoints along with the traditional taxonomical parameters in a perspective which can greatly enhance the evaluation of fluvial ecosystem quality for biomonitoring practices.

Exploring the status of motility, lipid bodies, deformities and size reduction in periphytic diatom community from chronically metal (Cu, Zn) polluted waterbodies as a biomonitoring tool.

Taxonomic metrics of diatoms are regularly used for aquatic biomonitoring, including testing for heavy metal stress. In contrast, non-taxonomical parameters in diatoms are rarely assessed. In the present study, taxonomical features of diatoms, such as cell density, chlorophyll a, species richness, and the Shannon index, were reduced at severely polluted (Cu, Zn) sites compared with less polluted sites. Some non-taxonomic parameters, such as, lipid bodies (LBs) number and size, carotenoid/chlorophyll a ratios, and frustule deformities were elevated at the severely polluted sites in comparison to the less polluted sites in both the areas. Cell size diminished and motility changed from smooth to erratic with increasing Cu and Zn pollution. Some of these behavioral and physiological changes were easily assessed (e.g., motility and formation of LBs), while morphological alterations (cell wall deformities and changes in cell size) requires more time and human expertise in diatom taxonomy. These parameters were consistent across metal concentrations of sediments, in the water, and in cells. The results illustrate the usefulness of these non-taxonomic parameters in biomonitoring, especially as early warning tools for ecotoxicity assessment and testing for sublethal effects. Some of these parameters, such as cell size and cell wall deformities, can be easily incorporated into traditional protocols, although LBs and motility metrics will require more effort.

Evaluation of various isotherm models, and metal sorption potential of cyanobacterial mats in single and multi-metal systems.

Isotherm curves for the biosorption of Cu(II), Cd(II) and Pb(II) by the biomass of five different cyanobacterial mats (Mat # 1-5) showed concave shape and plateau. Suitability of ten different isotherm models was evaluated for the equilibrium modeling of these isotherm curves, however, only the Toth model was found appropriate. Mat # 2, dominated by Phormidium sp., was identified as an excellent metal biosorbent because: (i) the Toth estimated maximum biosorption capacity (mmol g(-1)) of Mat # 2 for Pb(II) (1.028), Cu(II) (0.696) and Cd(II) (0.549) was the highest among the tested mats and compares favourably with Langmuir estimated metal sorption capacity of many seaweeds, regarded as the best metal biosorbents, (ii) Na+, K+ and Ca2+ did not substantially inhibit the biosorption of the test metals, (iii) and total metal sorption ability of Mat # 2 increased or remained unaffected in binary and ternary metal systems.

Sorption of paraquat and 2,4-D by an Oscillatoria sp.-dominated cyanobacterial mat.

The present study characterises sorption of two pesticides, namely, paraquat (PQ) and 2,4-dichlorophenoxyacetic acid (2,4-D) by an Oscillatoria sp.-dominated cyanobacterial mat. Sorption of PQ onto the test mat was not significantly affected by the pH of the solution within the pH range 2-7. However, 2,4-D sorption was strongly influenced by the solution pH and was maximum at pH 2. Whereas PQ sorption increased with increase in temperature, 2,4-D sorption showed an opposite trend. The sorption of PQ and 2,4-D achieved equilibrium within 1 h of incubation, independent of concentration of pesticide and mat biomass in the solution. The pseudo-second-order kinetic model better defined PQ sorption than the pseudo-first-order model, whereas 2,4-D sorption was well defined by both the models. Sorption isotherms of both the pesticides showed L-type curve. Freundlich model more precisely defined PQ sorption than Langmuir model, thereby suggesting heterogeneous distribution of PQ binding sites onto the biomass surface. However, the Langmuir model more correctly defined 2,4-D sorption, thus, indicating homogeneous distribution of 2,4-D binding sites onto the biomass surface. The test biomass is a good sorbent for the removal of PQ because it could, independent of pH of the solution, sorb substantial amount of PQ (q(max) = 0.13 mmol g(-1)).