Fischeri Bioluminescence Research Articles

Assessment of the Ecotoxicity of Nanoplastics

The presence of micro- and nanoplastics in aquatic environments (including freshwater and marine ecosystems as well as their sediments) is becoming an increasingly serious problem worldwide. A wide range of studies have addressed the ecological effects these particles pose on biota. The main exposure pathway are food chains, e.g. under laboratory conditions these particles accumulate in the brain tissues of fish that feed on zooplankton causing brain damage. These studies, however, report mainly on the physical effects. In order to establish actual ecotoxicological effects, nanoplastics (50 nm in diameter) were assessed using the Vibrio fischeri bioluminescence inhibition bioassay (VFBIA). Our results showed that even environmentally relevant concentrations might trigger ecotoxicological effects. This study can be con- sidered to be a first screening, however, results indicate the need for more complex testing on a battery of aquatic test organisms.

Biodegradation of PCBs in contaminated water using spent oyster mushroom substrate and a trickle-bed bioreactor

Due to their persistence, polychlorinated biphenyls (PCBs) represent a group of important environmental pollutants, but conventional physicochemical decontamination techniques for their removal are usually expensive. The main aim of this work was to develop a cost-effective method for PCB bioremediation, focusing on contaminated water and utilizing the well-known degradation capability of Pleurotus ostreatus (the oyster mushroom). For this purpose, the conditions of several laboratory-scale reactors (working volume 1 L) were optimized. Spent oyster mushroom substrate obtained from a commercial farm was used as a fungal inoculum and growth substrate. The highest degradation efficiency (87%) was recorded with a continuous low-flow setup, which was subsequently scaled up (working volume 500 L) and used for the treatment of 4000 L of real contaminated groundwater containing 0.1–1 μg/L of PCBs. This trickle-bed pilot-scale bioreactor was able to remove 82, 80, 65, and 30–50% of di-, tri-, tetra- and pentachlorinated PCB congeners, respectively. No degradation was observed for hexa- or heptachlorinated congeners. Multiple mono- and dichlorobenzoic acids (CBAs) were identified as transformation products by mass spectrometry, confirming the role of biodegradation in PCB removal. A Vibrio fischeri bioluminescence inhibition test revealed slight ecotoxicity of the primary reactor effluent (sampling after 24 h), which was quickly suppressed once the effluent passed through the reactor for the second time. Moreover, no other effluent exhibited toxicity for the rest of the experiment (71 days in total). Microbial analyses (phospholipid fatty acid analysis and next-generation sequencing) showed that P. ostreatus was able to degrade PCBs in the presence of an abundance of other fungal species as well as aerobic and anaerobic bacteria. Overall, this study proved the suitability of the use of spent oyster mushroom substrate in a bioremediation practice, even for pollutants as recalcitrant as PCBs.

Ecotoxicity, genotoxicity, and oxidative potential tests of atmospheric PM10 particles

The aim of the present work was to investigate the likely toxicological impact of atmospheric PM10 particles by comparing different effect-based methodologies, namely the Vibrio fischeri bioluminescence inhibition bioassay to evaluate ecotoxicity, the SOS Chromotest assay to estimate genotoxicity, and the Dithiothreitol (DTT) and Ascorbic Acid (AA) acellular assays to assess oxidative potential. The chemical composition was characterized for about 30 species, to assess the potential health impact of specific chemical components. Atmospheric particles were collected from spring to summer at a coastal site of the Central Mediterranean, away from large sources of local pollution. The Toxicity Unit (TU) index, used to assess the ecotoxicity, showed that 33% of the samples were toxic. The Induction Factor (IF), generally used to assess particle's genotoxicity, varied from 0.3 to 1.5 that represents the threshold value for genotoxicity. The oxidative potential (OP) determined by the DTT and AA assay varied within the 4.9–34.5 and 4.8–140.6 nmol min−1 range, respectively. DTT-OP and TU values were significantly correlated with OC, EC, and nss-K+, likely because the DTT and Vibrio fischeri responses were mainly associated with species from combustion sources. The IF factor was significantly correlated with some metals (Al, Ba, La, P, Sr, and Ti) likely from traffic sources and did not show any significant correlation with TU and OP values. Overall, paper's results proved the episodic occurrence of ecotoxicity and genotoxicity levels in PM10 particles sampled directly from their natural environment and away from strong pollution sources, highlighting the role mainly of carbonaceous compounds and heavy metals. The impact of spurious correlations between DTT- and AA-OPV and chemical species concentration has also been addressed.

Effect of nano-scale, reduced graphene oxide on the degradation of bisphenol A in real tertiary treated wastewater with the persulfate/UV-C process

In this study, the effect of home-made, reduced graphene oxide (rGO) nanoparticles on persulfate (PS)/UV-C oxidation of bisphenol A (BPA; 8.8 μM), an endocrine disruptor and model micropollutant, in real tertiary treated urban wastewater (TWW) was examined. Experimental results revealed that the presence of rGO induced a significant improvement in the degradation of BPA by the PS/UV-C treatment system. Fast and complete BPA removal accompanied with 44% dissolved organic carbon (DOC) abatement was achieved within 30 min at reaction conditions of 0.01 g/L rGO, 0.125 mM PS, at the natural pH of the TWW (pH = 7.5). This high performance on BPA degradation at the natural pH TWW could be attributable to the synergistic effect between the rGO nanoparticles and the PS/UV-C photochemical oxidation causing additional activation of PS and adsorption of BPA, DOC and degradation products onto the rGO surface. Radical quenching studies revealed that sulfate radicals prevailed over hydroxyl radicals. The Vibrio fischeri bioluminescence inhibition assay indicated that no toxic degradation products were formed during rGO/PS/UV-C treatment. By liquid chromatography time of flight mass spectrometry (LC-TOF-MS) analysis the formation of thirteen degradation products during PS/UV-C and rGO/PS/UV-C treatment of BPA was confirmed. This study underlined that the combined rGO/PS/UV-C treatment process could offer a sustainable, ecotoxicologically safe and kinetically attractive solution for the effective elimination of micropollutants from waters and wastewaters.

Influence of biochar particle size on biota responses

Despite the increasing interest for biochar as a soil amendment, a knowledge gap remains on its impacts on non-target soil and aquatic species. We hypothesised that biochar particle size and application rate can play a role in the toxicity to biota. Pine woodchip biochar was incorporated in a clean soil at three particle size classes: small (<0.5 mm), medium (1–2 mm), and large (<4 mm), and at two concentrations: 1% and 6% w/w. A laboratory screening with earthworm Eisenia andrei avoidance behaviour bioassay was carried out to test the most adequate application rates, particle sizes and soil-biochar pre-incubation period. Thereafter, a 28-day greenhouse microcosm experiment was conducted as an ecologically more representative approach. Survival, vertical distribution and weight changes of E. andrei, and bait-lamina consumption were recorded. Soil leachates from the microcosms were collected to evaluate their impact on Daphnia magna immobilisation and Vibrio fischeri (Microtox®) bioluminescence. A feeding experiment with E. andrei was also performed to address earthworm weight changes and to conduct a screening of PAH-type metabolites in their tissue. The 6% <0.5 mm treatment pre-incubated for 96 h induced significant avoidance of the earthworms. Significantly lower bait-lamina consumption was observed in microcosms for the 6% <0.5 mm treatment. Moreover, particle size was a statistically significant factor regarding the loss of weight in the feeding experiment and higher concentration of naphthalene-type metabolites detected in E. andrei tissue, when exposed to <0.5 mm biochar particles. The leachates had no adverse effects on the aquatic species. The results suggest that particles <0.5 mm of pine woodchip biochar can pose sub-lethal effects on soil biota.

Assessing Ecotoxicity of Size-fractionated Airborne Particulate Matter

Particulate matter (PM) is grouped as coarse, fine, and ultrafine particles (UFPs) with aerodynamic diameters of 2.5 to 10 μm (PM10), &lt;2.5 μm (PM2.5), and &lt;0.1 μm (PM0.1), respectively. The course and fine fractions have been well characterised from numerous aspects, including potential environmental hazard. However, more and more studies are targeted to the UFP fraction, as they bind relatively higher concentrations of potentially toxic materials and they might penetrate through cell biological barriers, posing higher risk to the biota. In our study, ecotoxic potential of size-fractionated urban aerosol was evaluated, using the kinetic version of the Vibrio fischeri bioluminescence inhibition bioassay. The kinetic protocol makes it possible to avoid false ecotoxicity readings which might appear in case of coloured and/or turbid samples. Our results showed that all PM fractions elucidated significant toxic response, highest toxicity was experienced in the range of 0.25/0.5μm and 0.5/1 μm (with the EC50s of 7.07 and 7.8%). Ecotoxicity in general followed the typical pattern of number size distributions of submicron particles experienced in Europe.

Redox mediators and irradiation improve fenton degradation of acesulfame

Widely recognized as a promising approach to degrading recalcitrant pollutants, Advanced Oxidation Processes (AOPs) have drawn much attention for their effectiveness and efficiency. Among all the AOPs, the Fenton system has been widely applied for oxidation and mineralization of micropollutants due to its ease of implementation and high catalytic efficiency. However, the necessity of preceding acidification, together with rapid consumption and slow regeneration of Fe(II) resulting in deterioration of reactivity, has reduced its competitiveness as a practical option for water treatment. Acknowledging the above drawbacks, this study investigates the potential viable option to enhance the Fenton system. Acesulfame was chosen as the model compound due to its ubiquitous occurrence and persistence in the environment. UV-assisted photo-Fenton treatment was found to remove the parent compound effectively; the transformation profile of acesulfame was identified and elucidated with the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Prolonged UV photo-Fenton treatment was effective for mineralization of the majority of the transformation products, without increasing the overall toxicity as indicated by Vibrio fischeri bioluminescence assay. The positive effects of the addition of redox mediators to Fenton systems at neutral pH were confirmed in this study. The results could be the basis for further development of homogeneous catalytic degradation techniques for the oxidation of environmental contaminants at circumneutral pHs to neutral pHs.

Treatment of aqueous solutions of 1,4-dioxane by ozonation and catalytic ozonation with copper oxide (CuO)

ABSTRACT In this study, treatment for the removal of 1,4-dioxane by ozone and by catalytic ozonation using CuO as the catalyst was investigated. While the removal of 1,4-dioxane was small (20%) and mineralization negligible after 6 h of ozonation treatment, the removals of 1,4-dioxane and total organic carbon increased by factors of 10.35 and 81.25, respectively, after catalytic ozonation in the presence of CuO. The mineralization during catalytic ozonation was favoured at pH 10 (94.91 min−1), although it proceeded even at pH 3 (54.41 min−1). The CuO catalyst decreased the equilibrium concentration of soluble ozone and favoured its decomposition to reactive oxidative species. Radical scavenging experiments demonstrated that superoxide radicals were the main species responsible for the degradation of 1,4-dioxane. Further scavenging experiments with phosphate confirmed the presence of Lewis active sites on the surface of CuO, which were responsible for the adsorption and decomposition of ozone. The reaction mechanism proceeded through the formation of ethylene glycol diformate, which quickly hydrolyzed to ethylene glycol and formic acid as intermediate products. The stability of CuO indicated weak copper leaching and high catalytic activity for five recycling cycles. The toxicity of the water, assessed by Vibrio fischeri bioluminescence assays, remained the same (low toxicity) after catalytic ozonation while it increased after treatment with ozonation alone.

Insights into influencing factor, degradation mechanism and potential toxicity involved in aqueous ozonation of oxcarbazepine (CHEM46939R1)

Oxcarbazepine (OXC), as a potent antiepileptic drug, is widely used in recent years, but its residue is potentially harmful to the environment. Although ozonation is a high-efficient technology for chemical oxidation during water treatment, it cannot completely mineralize organic matters, but partially transforms them into some unidentified by-products. In order to provide more insight into OXC ozonation process, the influencing factor, transformation mechanism and potential toxicity were comprehensively investigated in this study. The results showed that the optimal ozonation temperature was 20 °C with a pseudo-first-order reaction rate constant of 0.161 min−1. The increase of pH significantly enhanced OXC degradation, while the presence of bicarbonate caused a remarkable negative effect, manifesting that hydroxyl radical (OH) oxidation should play an important role in OXC ozonation. Moreover, transformation mechanism was further elucidated based on the identification of ten OXC-related by-products using UPLC-Q-TOF-MSn, which mainly consisted of electrophilic substitution, N-heterocyclic ring cleavage and re-arrangement, hydroxylation, carbonylation, demethoxylation and deamidation, etc. The toxicity evaluation, using US Environmental Protection Agency Toxicity Estimation Software Tool (US-EPA TEST), suggested that most identified by-products were probably more toxic than OXC itself. Besides, further experiments, by measuring inhibitory effect of ozonated mixture on Vibrio fischeri bioluminescence, demonstrated that by-products with higher toxicity tended to be accumulated under a short reaction time. Taken together, the present investigation provided valuable information for further understanding OXC ozonation process, and suggested that special attention should be paid to the control and elimination of toxic transformation by-products in future studies.

Particle Size and Concentration Dependent Ecotoxicity of Nano- and Microscale TiO2 —Comparative Study by Different Aquatic Test Organisms of Different Trophic Levels

A comprehensive ecotoxicity assessment of three different nanosized TiO2 (with 16, 36 and 89 nm particle diameter) and one microscale TiO2 suspension (with 3264 nm particle diameter) was carried out with a special emphasis on the relation between product characteristics and toxic effect. The applied test battery included the combination of modified standardized tests (Aliivibrio fischeri bioluminescence inhibition test, Lemna minor growth inhibition test), and nonstandardized bioassays with unconventional physiological endpoints (Tetrahymena pyriformis phagocytic activity, the Daphnia magna heartbeat rate). Based on the lowest significant effect values, the tested aquatic organisms were the most sensitive to the microscale TiO2 suspension (with 3264 nm particle size). Although the three nanoscale TiO2 particles were aggregated in the A. fischeri and the L. minor growth media, significant inhibition rates were experienced at 0.1 and at 1 μg L─1 concentration of nTiO2 suspensions with 16 and 36 nm primary particle size, respectively. Larger aggregates may have also high impact on biological organisms. In case of the D. magna heartbeat rate test rapid agglomeration was avoided, but lower responses were found compared to other investigated systems. The short term T. pyriformis phagocytic activity test demonstrated outstanding sensitivity; three TiO2 suspensions were significantly toxic even at 0.1 μg L─1. The consequences of our study clearly indicated that nanoscale TiO2 may have an impact on the aquatic ecosystem which is strongly influenced by aggregation. The effect of exposure duration and concentration as contributing factors in nano-titanium dioxide mediated toxicity was also demonstrated.

Acute toxicity of emerging atmospheric pollutants from wood lignin due to biomass burning

Guaiacol (2-methoxyphenol) is an important atmospheric pollutant. It is the major component of wood lignin and is essentially emitted to the atmosphere during biomass burning. Its aging in the tropospheric aqueous phase leads to the generation of the following ring-retaining transformation products, also during nighttime: 4-nitroguaiacol, 6-nitroguaiacol, and dinitroguaiacol. This study presents the first toxicological data of guaiacol and its nitro derivatives and reveals their harmful potential for the ecosystem. Applying V. fischeri bioluminescence acute toxicity test, EC50 values range from 16.7 to 103mgL−1 after a 30-min incubation period, which classifies all investigated compounds as ‘harmful’ according to the European legislation. The investigation of environmentally relevant mixtures did not show significant joint actions between the four studied compounds. Therefore, their concentration addition can be considered for ecotoxicological purposes. However, a synergistic effect between guaiacol and a minor unidentified first-generation product of its aqueous-phase aging was observed and should be taken into account when assessing the reaction mixture toxicity. These results stress the need for further toxicological testing, including organisms of different trophic levels, to better evaluate the environmental hazard of guaiacol and especially its nitro derivatives.

Water Quality and Ecotoxicity Assessment in Surface Waters from Cubat&amp;#227;o River and Surroundings, S&amp;#227;o Paulo, Brazil

The monitoring of water bodies means the attempt of protecting vulnerable groups of organisms inhabiting streams and rivers. Industrial and domestic discharges may worsen the water quality and affect biological balance, structure and the functioning of the ecosystem. Cubatão City, is one of the largest industrial centers in Brazil and in Latin America, where the constant discharge of effluents into Cubatão River and its tributaries caused a degradation scenario in the hydrographic basin of the region. The objective of this study was to evaluate the acute and chronic effects of surface water from Cubatão River and two of its tributaries (Perequê and Pilões) by ecotoxicological assays. In addition, physical chemical parameters were analyzed. Ceriodaphnia dubia and Vibrio fischeri were exposed-organisms during the studied period. The study was conducted between 2010 and 2011 in four campaigns and nine sites along the basin of Cubatão River. The ecotoxicity was measured by Vibrio fischeri bioluminescence, EC50 values ranging from 31.25% to 71.61%. In contrast, based on a bioequivalence t-test statistical analysis, the results obtained with Ceriodaphnia dubia revealed no toxicity in the sampling sites. A critical analysis of raw data of luminescence was carried out showing higher values during the 2nd campaign. From the numbers obtained for physical-chemical parameters P5 was far the worst due to chlorides, hardness and conductivity. From the Pearson correlation analysis carried out with toxicity to V. fischeri and the physical chemical parameters, the dissolved oxygen in water resulted in a moderate positive correlation. Sediment contamination was also demonstrated in the region.

Effect of Elderberry (Sambucus nigra L.) Extract Supplementation in STZ-Induced Diabetic Rats Fed with a High-Fat Diet.

Elderberry (Sambucus nigra L.) lipophilic and polar extract dietary supplementation effects were evaluated according to diabetes management indices, using an in vivo model. A research pipeline was constructed, that ranged from extract preparation, partial chemical characterization and toxicity evaluation, to examining the elderberry extract dietary supplementation effects on biofluid and tissues. Extracts toxicity was screened using an Aliivibrio fischeri bioluminescence model. A concentration of up to 60 mg/L was selected, and rat doses for oral supplementation were computed applying the interspecies correlation between A. fischeri and rats. Wistar type 2 diabetic rats, induced by streptozotocin (STZ), were fed a high-fat diet and supplemented for 4 weeks at doses of 190 and 350 mg/kg body weight/day of lipophilic and polar extract, respectively. As far as we know, lipophilic elderberry extract supplementation was assessed for the first time, while polar extract was administrated at higher doses and for a shorter period compared to previous studies, aiming to evaluate subacute supplementation effects. The polar extract modulated glucose metabolism by correcting hyperglycemia, while the lipophilic extract lowered insulin secretion. Both extracts lowered insulin resistance, without remarkable alterations to hematological indices, sera lipids and sera and tissular trace element homeostasis. In conclusion, elderberries are a potential source of bioactive compounds for formulations to be used as co-adjuvants in diabetes management.

Multilaboratory evaluation of 15 bioassays for (eco)toxicity screening and hazard ranking of engineered nanomaterials: FP7 project NANOVALID

Within EU FP7 project NANOVALID, the (eco)toxicity of 7 well-characterized engineered nanomaterials (NMs) was evaluated by 15 bioassays in 4 laboratories. The highest tested nominal concentration of NMs was 100 mg/l. The panel of the bioassays yielded the following toxicity order: Ag > ZnO > CuO > TiO2 > MWCNTs > SiO2 > Au. Ag, ZnO and CuO proved very toxic in the majority of assays, assumingly due to dissolution. The latter was supported by the parallel analysis of the toxicity of respective soluble metal salts. The most sensitive tests/species were Daphnia magna (towards Ag NMs, 24-h EC50 = 0.003 mg Ag/l), algae Raphidocelis subcapitata (ZnO and CuO, 72-h EC50 = 0.14 mg Zn/l and 0.7 mg Cu/l, respectively) and murine fibroblasts BALB/3T3 (CuO, 48-h EC50 = 0.7 mg Cu/l). MWCNTs showed toxicity only towards rat alveolar macrophages (EC50 = 15.3 mg/l) assumingly due to high aspect ratio and TiO2 towards R. subcapitata (EC50 = 6.8 mg Ti/l) due to agglomeration of TiO2 and entrapment of algal cells. Finally, we constructed a decision tree to select the bioassays for hazard ranking of NMs. For NM testing, we recommend a multitrophic suite of 4 in vitro (eco)toxicity assays: 48-h D. magna immobilization (OECD202), 72-h R. subcapitata growth inhibition (OECD201), 30-min Vibrio fischeri bioluminescence inhibition (ISO2010) and 48-h murine fibroblast BALB/3T3 neutral red uptake in vitro (OECD129) representing crustaceans, algae, bacteria and mammalian cells, respectively. Notably, our results showed that these assays, standardized for toxicity evaluation of “regular” chemicals, proved efficient also for shortlisting of hazardous NMs. Additional assays are recommended for immunotoxicity evaluation of high aspect ratio NMs (such as MWCNTs).

Application of advanced oxidation technologies and sand filter for the detoxification of effluents from small textile industries in Ecuador

AbstractFenton and photo-Fenton processes were tested for the treatment of wastewater from a local textile industry in Ecuador. The goal of this research was to obtain an efficient, easy-to-apply method for the removal of colour, organic compounds, biodegradability, resistant pollutants, and toxicity according to the Vibrio fischeri bioluminescence test. The best results for Fenton were obtained with 100 mM H2O2 and 0.36 mM Fe2+. Toxicity was reduced by 84%, biodegradability was increased from 0.21 to 0.6, and the resulting COD and BOD5 concentrations met the Ecuadorian legislation. Both temperature and UV radiation improved Fenton degradation. Detoxification was accomplished with a combination of sand filter with Fenton.

Comparison of organics and heavy metals acute toxicities to Vibrio fischeri

Vibrio fischeri bioluminescence inhibition has been widely used to test acute toxicities of metals and organics contaminants. However, the differences of metals and organics acute toxicities to V. fischeri have not been compared. Here, four heavy metals (Zn2+, Cu2+, Cd2+, Cr6+) and five organics (phenol, benzoic acid, p-hydroxy benzoic acid, nitro-benzene and benzene) acute toxicities to V. fischeri were investigated. Heavy metals toxicities to V. fischeri were increased along with the reaction time, while the organics toxicities kept the same level in different reaction times. In order to explain the difference, the relative cell death rate of V. fischeri was detected. In metals toxicities tests, the bioluminescence inhibition rate of V. fischeri was found to be significantly higher than the relative cell death rate (P&lt;0.05), while for the organics toxicities tests, the cell death rate was similar to the bioluminescence inhibition rate. These results indicated that organics acute toxicities to V. fischeri could reflect the death of cell, but metals acute toxicities to V. fischeri may not lead to the death of cell, just represent the bioluminescence inhibition.

The influence of extreme river discharge conditions on the quality of suspended particulate matter in Rivers Meuse and Rhine (The Netherlands)

As a consequence of climate change, increased precipitation in winter and longer periods of decreased precipitation in summer are expected to cause more frequent episodes of very high or very low river discharge in the Netherlands. To study the impact of such extreme river discharge conditions on water quality, toxicity profiles and pollutant profiles were determined of suspended particulate matter (SPM) collected from Rivers Meuse and Rhine. Archived (1993–2003) and fresh (2009–2011) SPM samples were selected from the Dutch annual monitoring program of the national water bodies (MWTL), representing episodes with river discharge conditions ranging from very low to regular to very high. SPM extracts were tested in a battery of in vitro bioassays for their potency to interact with the androgen receptor (AR), the estrogen receptor (ER), the arylhydrocarbon receptor (AhR), and the thyroid hormone transporter protein transthyretin (TTR). SPM extracts were further tested for their mutagenic potency (Ames assay) and their potency to inhibit bacterial respiration (Vibrio fischeri bioluminescence assay). Target-analyzed pollutant concentrations of the SPM samples and additional sample information were retrieved from a public database of MWTL results. In vitro toxicity profiles and pollutant profiles were analyzed in relation to discharge conditions and in relation to each other using correlation analysis and multivariate statistics. Compared to regular discharge conditions, composition of SPM during very high River Meuse and Rhine discharges shifted to more coarse, sandy, organic carbon (OC) poor particles. On the contrary, very low discharge led to a shift to more fine, OC rich material, probably dominated by algae. This shift was most evident in River Meuse, which is characterized by almost stagnant water conditions during episodes of drought. During such episodes, SPM extracts from River Meuse demonstrated increased potencies to inhibit bacterial respiration and to compete with thyroid hormone to bind to TTR, possibly due to the presence of fycotoxins. Meanwhile concentrations of polychlorobiphenyls (PCBs) in SPM were also increased. Very high River Meuse discharges on the other hand corresponded to increased androgenic and AhR agoniztic responses, which coincided with increased PAH levels and PAH-related in vivo risk estimates (i.e. multi-substance potentially affected fraction of species; msPAF). In River Rhine, very high discharges also corresponded to increasing androgenic potencies in SPM. Concentrations and corresponding msPAF values of PAHs (and metals), however, decreased with very high discharges in River Rhine in contrast to River Meuse. Mutagenicity was observed for SPM extracts from River Rhine collected during all discharge conditions, except during regular discharge. Aggregated toxicity index values, which were useful to identify toxicity profiles deviating from the generally observed pattern, did not correlate with river discharges, probably due to opposite effects of discharge conditions on different bioassay responses. In conclusion, SPM quality and related in vivo risk estimates changed during very low or very high discharge conditions but the changes were specific for the different toxic endpoints and pollutants in the different rivers. Moreover, bioassay responses to a series of consecutively collected samples from River Rhine during the Christmas flood of 1993 indicated that SPM quality is variable within a single episode of extreme discharge.

Toxicity of Some Pesticides, Heavy Metals and Their Mixtures to Vibrio fischeri Bacteria and Daphnia magna: Comparative Study

The present study was conducted to evaluate the sensitivity of the Vibrio fischeri bioluminescence inhibition test (Microtox® assay), and the standard acute Daphnia magna test; using 3 heavy metals, 3 organic pesticides, and their mixtures. In Daphnia tests, either at 24h or 30 min exposure times, the pattern of toxicity order for heavy metals was Cu ˃ Cd ˃ Pb. Chlorpyrifos-methyl was the highest toxic at 24h, while Triazophos was the highest toxic at 30 min exposure times. In the Microtox® test at 5 min exposure time, the estimated EC50 values were 4.20, 4.53 and 6.60 mg/L for Cu, Cd and Pb, respectively. At the same exposure time, the EC50 values of Triazophos, Chlorpyrifos-Me and Profenofos accounted to 1.76, 3.36 and 4.12 mg/L, respectively. Similar order of toxicity was obtained when tests were conducted at 15 min exposure time. The paired mixtures of pesticides, as well as the mixtures of Cu + Cd and Pb + Cd, showed potentiation effects, while the mixture of Cu + Pb showed additive effect against D. magna. The tertiary mixtures of the pesticides or the heavy metals reacted antagonistically. In the Microtox® assay, the heavy metal mixtures reacted antagonistically, while pesticide mixtures showed synergism. It was concluded that both Daphnia and Microtox® tests showed similar pattern of sensitivity to the single toxicants, but dissimilar pattern to the heavy metal mixtures. On the other side, using shorter exposure time (ca. 30 min) with Daphnia bioassay may enable us to held reliable comparisons with Microtox® results.

Novel linear polymers able to inhibit bacterial quorum sensing.

Bacterial phenotypes, such as biofilm formation, antibiotic resistance and virulence expression, are associated with quorum sensing. Quorum sensing is a density-dependent regulatory system of gene expression controlled by specific signal molecules, such as N-acyl homoserine lactones (AHLs), produced and released by bacteria. This study reports the development of linear polymers capable to attenuate quorum sensing by adsorption of AHLs. Linear polymers were synthesized using MMA as backbone monomer and methacrylic acid and itaconic acid as functional monomers. Two different quorum sensing-controlled phenotypes, Vibrio fischeri bioluminescence and Aeromonas hydrophila biofilm formation, were evaluated to test the polymers' efficiency. Results showed that both phenotypes were significantly affected by the polymers, with the itaconic acid-containing material being more effective than the methacrylic acid one. The polymer inhibitory effects were reverted by the addition of lactones, confirming attenuation of quorum sensing through sequestration of signal molecules. The polymers also showed no cytotoxicity when tested using a mammalian cell line.

Weight-of-evidence approach in assessment of ecotoxicological risks of acid sulphate soils in the Baltic Sea river estuaries

Acidity and leaching of metals from acid sulphate soils (ASSs) impair the water quality of receiving surface waters. The largest ASS areas in Europe are found in the coasts of the northern Baltic Sea. We used weight-of-evidence (WoE) approach to assess potential risks in 14 estuary sites affected by ASS in the Gulf of Finland, northern Baltic Sea. The assessment was based on exposure and effect profiles utilizing sediment and water metal concentrations and concurrent pH variation, sediment toxicity tests using the luminescent bacterium Vibrio fischeri and the midge Chironomus riparius, and the ecological status of benthic macroinvertebrate communities. Sediment metal concentrations were compared to national sediment quality criteria/guidelines, and water metal concentrations to environmental quality standards (EQSs). Hazard quotients (HQs) were established for maximum aluminium, cadmium and zinc concentrations at low pH based on applicable US EPA toxicity database. Sediment metal concentrations were clearly elevated in most of the studied estuaries. The EQS of cadmium (0.1μg/l) was exceeded in 3 estuaries out of 14. The pH-minima were below the national threshold value (5.5) between good and satisfactory water quality in 10 estuaries. V. fischeri bioluminescence indicated toxicity of the sediments but toxic response was not observed in the C. riparius emergence test. Benthic invertebrate communities were deteriorated in 6 out of 14 sites based on the benthic invertebrate quality index. The overall ecotoxicological risk was assessed as low in five, moderate in three and high in five of the estuary sites. The risk assessment utilizing the WoE approach indicated that harmful effects of ASSs are likely to occur in the Baltic Sea river estuaries located at the ASS hotspot area.