Phytochemical Profile and Evaluation of the Antioxidant, Cyto-Genotoxic, and Antigenotoxic Potential of Salvia verticillata Hydromethanolic Extract.

This study comprises the phytochemical characterization, the evaluation of the total phenolic content (TPC) and antioxidant activity (AA), and the investigation of the cyto-genotoxic and antigenotoxic potential of hydromethanolic extract derived from Salvia verticillata L. leaves. HPLC-DAD-ESI-MS and HPLC-DAD were used for the characterization of the extract and determination of the major ingredients. Afterwards, the TPC and AA were determined. The cytotoxic and genotoxic effect of the extract on cultured human lymphocytes at concentrations of 10, 25, and 50 µg mL-1 was investigated via the Cytokinesis Block MicroNucleus (CBMN) assay. Moreover, its antigenotoxic potential against the mutagenic agent mitomycin C (MMC) was assessed using the same assay. The hydromethanolic extract comprises numerous metabolites, with rosmarinic acid being the major compound. It had a high value of TPC and exerted significant AA as shown by the results of the Ferric Reducing Antioxidant Power (FRAP) and Radical Scavenging Activity by DPPH• assays. A dose-dependent cytotoxic potential was recorded, with the highest dose (50 µg mL-1) exhibiting statistically significant cytotoxicity. None of the tested concentrations induced significant micronuclei (MN) frequencies, indicating a lack of genotoxicity. All tested concentrations reduced the MMC-mediated genotoxic effects, with the two lowest showing statistically significant antigenotoxic potential.

New insights into the size-independent bioactive potential of pristine and UV-B aged polyethylene microplastics.

The present study investigates the morphological, physicochemical, and structural changes occurred by the UV-B aging process of low-density polyethylene microplastics (LDPE MPs), as well as the bioactive potential of both pristine and UVaged MPs towards healthy peripheral blood lymphocytes. Specifically, LDPE MPs (100-180 µm) prepared by mechanical milling of LDPE pellets, were UV-B irradiated for 120 days (wavelength 280 nm; temperature 25 °C; relative humidity 50 %) and further examined for alterations in their particle size and surface, their functional groups, thermal stability, and crystallinity (by means of SEM, FTIR spectroscopy, XRD patterns, and TGA measurements, respectively). In parallel, isolated human peripheral blood lymphocytes were treated with different concentrations (25-500 µg mL-1) of either pristine or aged MPs (UVfree and UV120d LDPE MPs) for assessing the cytogenotoxic (by means of trypan blue exclusion test and the cytokinesis-block micronucleus assay using cytochalasin-B) and oxidative effects (using the DCFH-DA staining) in both cases. According to the results, UVfree and UV120d-LDPE MPs, with a size ranging from 100 to 180 µm, can differentially promote cytogenotoxic and oxidative alterations in human lymphocytes. In fact, UVfree LDPE MPs not being able to be internalized by cells due to their size, could indirectly promote the onset of mild oxidative and cytogenotoxic damage in human peripheral lymphocytes, via a dose-dependent but size-independent manner. The latter is more profound in case of the irregular-shaped UV120d-LDPE MPs, bearing improved dispersibility and sharp edges (by means of cracks and holes), as well as oxygen-containing and carbonyl groups. To our knowledge, the present findings provide new data regarding the bioactive behavior of pristine and UV-B aged LDPE MPs, at least in the in vitro biological model tested, thus giving new evidence for their size-independent and/or indirect mode of action.

Different isoforms of parabens into marine environment: Biological effects on the bacterium Aliivibrio fischeri and the marine mussel Mytilus galloprovincialis.

Different isoforms of alkyl esters of p-Hydroxybenzoic acid, also known as parabens, are of great concern due to their widespread presence into the aquatic environment, their high concentrations in wastewater discharges, as well as their ability to induce adverse effects on aquatic organisms. Considering the imperative need for assessing their fate and risk to aquatic environment, the present study investigated the biological effects of two isoforms of parabens, methyl- (MeP) and propyl- (PrP), on the bacterium Aliivibrio fischeri (using the Bioluminescence Inhibition/Microtox® bioassay) and the mussel Mytilus galloprovincialis (in terms of mussel mortality, cellular, oxidative and genotoxic stress indices). The assessment of MeP and PrP behavior into aquatic media (artificial sea water/ASW and 2 % NaCl), primarily performed by UHPLC-UV-MS analysis, showed only a slight hydrolysis of PrP to 4-Hydrobenzoic acid (4-HBA). Furthermore, exposure of both species to different concentrations of each paraben revealed differences among their toxic potential, as well as their ability to cause cellular, oxidative and genotoxic effects on hemocytes of challenged mussels. Interestingly, the Microtox® bioassay showed that PrP mediated toxicity in A. fischeri were more pronounced than MeP, as revealed by the estimated toxic endpoints (in terms of concentration that promote 50 % of bioluminescence inhibition, EC50). Similarly, in challenged mussels, a significant disturbance of mussel hemocytes' lysosomal membrane integrity, as well as enhanced levels of superoxides, nitric oxides, lipid peroxidation byproducts, and micronuclei formation were observed. These findings are of great interest, since MeP and PrP differential toxic potential, as well their ability to induce pre-pathological alterations in marine species, like mussels, give new evidence for their risk to aquatic biota.

Phytochemical Analysis and Genotoxicological Evaluation of Prickly Pear Peel Extracts.

This study investigated the beneficial properties of prickly pear peel (PPP) extracts from Opuntia ficus-indica (L.) Mill. Extracts were obtained via the Soxhlet extraction method using methanol (P1), ethanol (P2) and ethanol-water (P3) as extraction solvents. Their total phenolic and flavonoid content (TPC and TFC, respectively) and their antioxidant activity (AA) were determined. The PPP extracts were characterized in detail using mass spectrometry techniques. Their cyto-genotoxic effect and antigenotoxic potential against mitomycin C were evaluated via the cytokinesis block micronucleus (CBMN) assay on human lymphocytes. Enhanced TPC, TFC and AA values were recorded for all the extracts. Moreover, P1 and P2 were cytotoxic only at the highest concentrations, whereas P3 was found to be cytotoxic in all cases. No significant micronucleus induction was observed in the tested extracts. The PPP extracts contain bioactive compounds such as flavonoids, carboxylic acids, alkaloids, fatty acids and minerals (mainly K, Si, Mg, Ca, P and Zn). The results showed that all three extracts exerted high antigenotoxic activity. Our findings confirm the beneficial and genoprotective properties of PPP extracts and further studies on the bioactive compounds of Opuntia ficus-indica (L.) Mill. are recommended, as it constitutes a promising plant in pharmaceutical applications.

Beneficial properties of Drimia numidica leaf methanolic extract against the cytogenotoxic effects of mitomycin C on human lymphocytes.

This study investigated the phytochemical profile of Drimia numidica leaf methanolic extract, as well as its cyto-genotoxic and cyto/genoprotective potential against mitomycin C (MMC) mediated effects on healthy human lymphocytes. Photosynthetic pigments, trace elements, and secondary metabolites were estimated and/or identified in methanolic extract of mature leaves, and the latter was further used for assessing its in vitro biological effects on MMC-free and/or MMC-treated human lymphocytes (at low, non-toxic concentrations of 0.001 and 0.01% v/v). The results showed that D. numidica leaf methanolic extract, being rich in carotenoids, phenolics, flavonoids, organic acids and bufadienolides, could be protective against MMC mediated cyto/genotoxic potential in healthy human lymphocytes. Biomolecules possessing antioxidant and antitumor potential, such as beta-carotene and lutein among others, chlorogenic acid, caffeic acid and their derivatives, minerals such as Si, as well as apigenin- and luteolin-derived glycosides, either individual or in a mixture, could be beneficial rather than harmful, at least at the extract concentrations tested. Although further in vitro and in vivo studies are still needed for elucidating the beneficial (individual and/or additive/synergistic) role of those compounds, the results of the present study are quite promising, thus encouraging new challenges for the appropriate utilization of D. numidica leaf extract.

Printing Ink Wastewater Treatment Using Hydrodynamic Cavitation and Coagulants/Flocculants.

Raw printing ink wastewater (PIW) was treated with various inorganic coagulants and organic flocculants (anionic and cationic polyacrylamides). These processes were also examined as post treatment step following hydrodynamic cavitation. Treatment effectiveness was assessed through color, chemical oxygen demand (COD) and total suspended solids (TSS) removal. The addition of 4500 mg L-1 polyaluminum chloride coagulant in undiluted PIW (COD: 17000 mg L-1) resulted in 99% color removal, 96% COD and TSS removal, after settling for 2 h. The addition of 10 mg L-1 of anionic polyacrylamides in the sample reduced settling time to only 5 min, with concomitant 96-98% removal efficiency. The addition of a 4 min hydrodynamic cavitation pretreatment step reduced coagulant addition by 33%, for the treatment of undiluted PIW (with 10 mg L-1 anionic polyacrylamide), while removals were ranged between 96 and 98%. Economic analysis for the undiluted PIW showed that costs were reduced by ca. 20% with the hydrodynamic cavitation pretreatment step. Moreover, sludge characterization showed the presence of maghemite, aluminum chloride and potassium aluminum silicate. Finally, toxicity tests revealed a significant attenuation of the toxic potential of undiluted PIW, thus indicating the enhanced efficiency of the proposed combined process (hydrodynamic cavitation and coagulation/flocculation).

Investigation of the Genotoxic, Antigenotoxic and Antioxidant Profile of Different Extracts from Equisetum arvense L.

The present study investigated the cyto-genotoxic and antigenotoxic effects of four different extracts of Equisetum arvense L. (common name: field horsetail) on human lymphocytes. Specifically, Soxhlet's prepared extracts from E. arvense L., using different solvents (S1: methanol (MeOH)-, S2: ethanol (EtOH)-, S3: water-, and S4: ethanol/water (EtOH-W)-) were analyzed for (a) their total phenolic and flavonoid content (TPC and TFC, respectively), (b) their antioxidant activity (AA), via the DPPH, FRAP and ABTS assays, and (c) their cyto-genotoxic and/or protective efficiency against the mutagenic agent mitomycin C, via the Cytokinesis Block MicroNucleus assay. All extracts showed increased TPC, TFC, and AA values in almost all cases. S1, S3 and S4 demonstrated no cytotoxic potential, whereas S2 was cytotoxic only at the highest concentrations. Genotoxicity was not observed in the tested extracts. The highest antigenotoxic activity was observed for EtOH-W (S4) extract, which was found to be rich in flavonoids, flavonoid-O-glycosides, phytosterols, phenolic and fatty acids as well as in minerals and mainly in K, Ca, Mg, Si and P, as assessed by using various mass spectrometry techniques. Those findings confirm that E. arvense L. extracts could be valuable candidates for medicinal applications and pharmaceutical products, thus alleviating the effects of more conventional drugs.

Printing ink wastewater treatment using combined hydrodynamic cavitation and pH fixation.

Printing ink wastewater (PIW) carries a heavy load of pollutants, the composition of which makes treatment difficult, especially when trying to minimize the pollution load. According to the latter, the present study aims to investigate PIW treatment with different various methods and to determine the maximum color, COD (chemical oxygen demand) and TSS (total suspended solids) removal. First, hydrodynamic cavitation (HC) was tested and the effect of hydrogen peroxide dosage (0-10 g L-1), and pH (3, 5, 8, 10) was examined concerning the removal of PIW initial COD concentrations 4000 and 8000 mg L-1. Removal was high (more than 81%) only at pH 5 in HC reactor. The second method involved treatment with separate pH fixation of the undiluted PIW (COD 17000 mg L-1, actual pH 8 ± 0.2). This treatment, maximized removals, reaching reduction of the initial values more than 91%, at pH 5. Finally, PIW was treated with a combination of the above methods, leading to 93-97% removals for 8000 mg L-1 PIW treatment and 97-99% for 17000 mg L-1 PIW respectively. Process cost calculations showed that the latter method is an effective and affordable treatment method for PIW streams, while toxicity tests of the treated PIW showed substantial toxicity reduction.

Chemical and biological tracking in decentralized sanitation systems: The case of artificial constructed wetlands.

Given that the social and economic sustainability of rural areas is highly based on the protection of natural resources, biodiversity and human health, simple-operated and cost-effective wastewater treatment systems, like artificial constructed wetlands (CWs), are widely proposed for minimizing the environmental and human impact of both water and soil pollution. Considering that the optimization of wastewater treatment processes is vital for the reduction of effluents toxic potential, there is imperative need to establish appropriate management strategies for ensuring CW performance and operational efficiency. To this end, the present study aimed to assess the operational efficiency of a horizontal free water surface CW (HFWS-CW) located in a world heritage area of Western Greece, via a twelve-month duration Toxicity Identification Evaluation (TIE)-like approach, including both chemical and biological tracking tools. Conventional chemical tracking, by means of pH, conductivity, total COD, and nitrogen-derived components, like nitrates and ammonia-nitrogen, were monthly recorded in both influents and effluents to monitor whether water quality standards are maintained, and to assess potent CW operational deficiencies occurring over time. In parallel, Whole Effluent Toxicity (WET) bioassays were thoroughly applied, using freshwater algae and higher plant species (producers), crustaceans and rotifers (consumers), as well as human lymphocytes (in terms of Cytokinesis Block Micronucleus assay) to evaluate the acute and short-term toxic and hazardous potential of both influents and effluents. The integrated analysis of abiotic (physicochemical parameters) and biotic (toxic endpoints) parameters, as well as the existence of "cause-effect" interrelations among them, revealed that CW operational deficiencies, mainly based on poorly removal rates, could undermine the risk posed by treated sewage. Those findings reinforce the usage of WET testing, thus giving rise to the importance of applying appropriate water management strategies and optimization actions, like oxygen enrichment of surface and bottom of HFWS-CW basins, expansion of the available land, the enhancement of bed depth and seasonal harvesting of plants, for ensuring sewage quality, in favor of water resources protection and sustainable growth in rural areas.

The Potential Risk of Electronic Waste Disposal into Aquatic Media: The Case of Personal Computer Motherboards.

Considering that electronic wastes (e-wastes) have been recently recognized as a potent environmental and human threat, the present study aimed to assess the potential risk of personal computer motherboards (PCMBs) leaching into aquatic media, following a real-life scenario. Specifically, PCMBs were submerged for 30 days in both distilled water (DW) and artificial seawater (ASW). Afterwards, PCMBs leachates were chemically characterized (i.e., total organic carbon, ions, and trace elements) and finally used (a) for culturing freshwater (Chlorococcum sp. and Scenedesmus rubescens) and saltwater (Dunaliella tertiolecta and Tisochrysis lutea) microalgae for 10 days (240 h), (b) as the exposure medium for mussel Mytilus galloprovincialis (96 h exposure), and (c) for performing the Cytokinesis Block Micronucleus (CBMN) assay in human lymphocytes cultures. According to the results, PCMBs could mediate both fresh- and marine algae growth rates over time, thus enhancing the cytotoxic, oxidative, and genotoxic effects in the hemocytes of mussels (in terms of lysosomal membrane impairment, lipid peroxidation, and NO content and micronuclei formation, respectively), as well as human lymphocytes (in terms of MN formation and CBPI values, respectively). The current findings clearly revealed that PCMBs leaching into the aquatic media could pose detrimental effects on both aquatic organisms and human cells.

PCB cause global DNA hypomethylation of human peripheral blood monocytes in vitro.

We have recently reported significant associations between exposure to polychlorinated biphenyls (PCB) and alterations on genome-wide methylation of leukocyte DNA of healthy volunteers and provided evidence in support of an etiological link between the observed CpG methylation variations and chronic lymphocytic leukemia. The present study aimed to elucidate the effects of PCB in human lymphocytes' methylome in vitro. Therefore, U937 cells and human peripheral blood monocytes (PBMC) were exposed in vitro to the dioxin-like PCB-118, the non-dioxin-like PCB-153, and hexachlorobenzene (HCB) and thorough cytotoxicity, genotoxicity and global CpG methylation analyses were performed. All compounds currently tested did not show any consistent significant genotoxicity at all exposure periods and concentrations used. On the contrary, extensive dose-dependent hypomethylation was observed, even at low concentrations, in stimulated PBMC treated with PCB-118 and PCB-153 as well as a small but statistically significant hypomethylation in HCB-treated stimulated cells.

Insights into the toxicity of biomaterials microparticles with a combination of cellular and oxidative biomarkers.

Considering that the extensive biomedical, pharmaceutics, cosmetic and other industrial applications of biomaterials (BMs) is of great concern nowadays, regarding their environmental risk, the present study aimed to investigate the effects of four BMs, poly(ε-caprolactone) (PCL), poly(butylene succinate) (PBSu), chitosan (CS) and modified chitosan (succinic acid grafted chitosan) (CS-Suc) in the form of microplastics (particle sizes less than 1 mm) on biochemical parameters of snails Cornu aspersum hemocytes. Due to the absence of knowledge about the environmentally relevant concentrations of BMs, snails were initially treated through their food with a wide range of nominal concentrations of each BM to define the half maximal effective concentration (NRRT50), according to the destabilization degree of hemocytes' lysosomal membranes (by mean of neutral red retention time/NRRT assay). Thereafter, snails were treated with each BM, at concentrations lower than the estimated NRRT50 values in all cases, for periods up to 15 days. After the end of the exposure period, a battery of stress indices were measured in hemocytes of challenged snails. According to the results, all parameters tested in BMs-treated snails statistically differed from those measured in BMs-free snails, thus indicating the pro-oxidant potential of BMs, as well as their ability to affect animals' physiology. The most considerable effect in most cases seems to be caused by modified chitosan and PCL, while chitosan appears to be the least toxic. A common response mechanism of snails' blood cells against the 4 BMs used in the present study was shown. After exposure to each of the studied BMs a significant augmentation in protein carbonyls, MDA equivalents and DNA damage, while a significant reduction in NRRT values was determined in the snails hemocytes, in relation to the unexposed animals. From the biochemical parameters examined, MDA equivalents and DNA damage seem to be more susceptible than the other parameters studied, to respond to BMs effect, with MDA to react with more sensitivity to PCL and CS, while DNA damage to CS-Suc and PBSu. Our results could suggest the simultaneous use of the latter biomarkers in biomonitoring studies of terrestrial ecosystems against the specific BMs.

ZnO, Ag and ZnO-Ag nanoparticles exhibit differential modes of toxic and oxidative action in hemocytes of mussel Mytilus galloprovincialis.

The present study investigates the cytotoxic and oxidative effects of custom-made nanoparticles (NPs) on hemocytes of Mytilus galloprovincialis, utilizing hemolymph serum (HS) as exposure medium. Specifically, hemocyte lysosomal membrane destabilization (in terms of neutral red retention time assay/NRRT), superoxide anion (O2-), nitric oxide (NO, in terms of nitrites) and lipid peroxidation content (in terms of malondialdehyde/MDA equivalents) were determined in cells treated for 1 h with different concentrations (0.1-50 µg mL-1) of ZnO NPs, Ag NPs and ZnO-Ag NPs, as well as AgNO3 and/or ZnCl2 (bulk ions, respectively). According to the results, Ag NPs were more cytotoxic than ZnO-Ag NPs and/or ZnO NPs, while NRRT values observed in AgNO3 treated cells were lower than those of ZnCl2. Furthermore, high levels of both O2- and MDA were detected in cells treated with Ag NPs, ZnO-Ag NPs, and AgNO3 at concentrations lower than 5 µg mL-1, while high NO generation was observed only in cells treated with 5-25 µg mL-1 of ZnO NPs or ZnCl2. Despite the absence of data, regarding the formation of NP-serum protein corona complexes that could mediate NP surface energy and uptake efficiency, the current study firstly revealed that ZnO NPs, probably via their surface charge, particle agglomeration, and NP Zn+ release could promote an immune-related generation of O2- and NO via the respiratory burst stimulation, a process that is questioned in the case of Ag NPs and/or ZnO-Ag NPs. Moreover, ZnO-Ag NP interaction with biological membranes and their oxidative mode of action seemed to be regulated by the release and the antagonistic/synergistic response of its ionic counterparts (ZnO+ and Ag+), but further studies are needed to elucidate the oxidative mode of action of NP metal ions in complex NP mixtures.

Physicochemical and Toxicological Assay of Leachate from Malt Spent Rootlets Biochar.

The aim of this study was to characterize the leachate derived from biochar produced from malt spent rootlets (MSR) and to evaluate the required washing level in order to provide water free from inorganic substances. MSR biochar was placed in a column and subjected to six serial washes with distilled water, and the leachate was analysed for main anions and heavy metals. The 1st wash aliquot contained increased levels of mainly phosphates (980 mg/L) and chlorides (760 mg/L), and lower levels of nitrates, sulfates, fluoride and bromide, which were decreased over washes. Zero concentrations were observed after three washes for most anions. The increased levels of Zn, Be, Cs, Mn, V and Se determined in the 1st wash aliquot were eliminated in the successive washes. The toxic potency of each wash aliquot, determined by the use of the fairy shrimp Thamnocephalus platyurus showed that the 1st and 2nd MSR biochar leachates were toxic with 4.52 and 1.46 toxic units (TU), respectively, followed by a significant elimination of toxicity after further washes.

Assessing the cyto-genotoxic potential of model zinc oxide nanoparticles in the presence of humic-acid-like-polycondensate (HALP) and the leonardite HA (LHA).

The present study investigates the potential cyto-genotoxic effects of model zinc oxide nanoparticles (ZnO NPs) on human lymphocytes, with and/or without humic acids (HAs). Two types of HAs were studied, a natural well-characterized leonardite HA (LHA) and its synthetic-model, a humic-acid-like-polycondensate (HALP). The Cytokinesis Block Micronucleus (CBMN) assay was applied in cell cultures treated with different concentrations of ZnO NPs (0.5, 5, 10, 20 µg mL-1) and under different concentrations of either HALP or LHA (ZnO NPs-HALP and ZnO NPs-LHA, at concentrations of 0.5-0.8, 5-8, 10-16, 20-32 and 0.5-2, 5-20, 10-40, 20-80 µg mL-1, respectively). According to the results, ZnO NPs lacked genotoxicity but demonstrated cytotoxic potential. Binary mixtures of ZnO NPs-HAs (ZnO NPs-HALP or ZnO NPs-LHA) showed negligible alterations of micronuclei (MN) formation in challenged cells, with cytotoxic effects revealed only in case of cells treated with ZnO NPs-LHA at the concentration 5-20 µg mL-1. Furthermore, no genotoxic phenomena were exerted neither by the ZnO NPs nor from their mixtures with HAs. These findings indicate [i] the cytotoxic activity of used ZnO NPs on human lymphocytes, and [ii] reveal the protective role of HAs against ZnO NPs mediated cytotoxicity.

Assessing the seasonal and intrinsic variability of neurotoxic and cyto-genotoxic biomarkers in blood of free-living Eleonoras' falcons.

In the present study we investigated seasonal and intrinsic variability of neurotoxic and cyto-genotoxic biomarkers in blood plasma and erythrocytes of free-living Eleonoras' falcons, captured during the pre-breeding (May of 2017 and 2018) and breeding period (September of 2017) on the Antikythira Island (Greece). Specifically, blood samples of captured birds were prepared for the determination of cholinesterase (ChEs, i.e. acetylcholinesterase/AChE and butyrylcholinesterase/ BChE) activity, as well as the formation of nuclear (i.e. the formation of micronuclei into the cells/MN, binucleated cells/BN and others), and cellular/cytoplasmic (i.e. echinocytes/EC, acanthocytes/AC and notched cells/NC) abnormalities in blood plasma and erythrocytes, respectively. Our results indicated that birds sampled in late May had higher ChE and BChE activity levels, as well as higher frequency of total nuclear abnormalities. The latter were also higher in second calendar year (2cy) birds. Cellular/cytoplasmic abnormalities were less frequent in falcons having better body condition, sampled in late May, as well as in light-morph falcons. The observed ChEs activities, as well as nuclear and cellular/cytoplasmic abnormalities revealed that Eleonora's falcons are likely to be exposed to chemical agents with neurotoxic and cyto-genotoxic potential year round, while different aspects of their biology and ecology, such as their reproductive and nutritional status, could mediate their levels. Although we encourage more sampling campaigns to verify the identified seasonal and intrinsic sources of variation in biomarkers tested, the current study enriches the existing knowledge about their usefulness in the environmental monitoring and risk assessment of migratory birds, like Eleonoras' falcon.

Feeding regimes modulate biomarkers responsiveness in mussels treated with diclofenac.

This study investigated the role of the feeding regime on cellular (lysosomal membrane impairment), oxidative (superoxides and nitric oxides generation, as well as lipid peroxidation) and genotoxic (nuclear abnormalities) biomarkers measured in hemocytes of mussels Mytilus galloprovincialis treated with diclofenac (DCF). Specifically, unfed mussels, or mussels fed ad libitum with algal species Tisochrysis lutea or Tetraselmis suecica (Tiso/DCF- and Tetra/DCF- treated mussels, respectively) were exposed to DCF (20 µgL-1) for 4 days. The results showed that biomarkers' responsiveness against DCF, were more pronounced in unfed and Tetra/DCF-, rather than Tiso/DCF- treated mussel hemocytes, thus revealing food deprivation, changes in mussel feeding/filtration rate and digestion processes, as potent factors of mussels' immune efficiency and response against DCF. Those findings could provide valuable data for the optimization of mussels' feeding regime during laboratory studies, in order to assess reliably the effects of emerging contaminants on non-target sentinel organisms, such as mussels.

Effects of Burkholderia thailandensis rhamnolipids on the unicellular algae Dunaliella tertiolecta.

The effects of rhamnolipids (RLs) produced and further purified from Burkholderia thailandensis, on the unicellular microalgae Dunaliella tertiolecta were investigated, in terms of RLs ability to affect algal growth, photosynthetic apparatus structure and energy flux, round and through photosystems II and I. Specifically, 24-48 h RLs-treated algae (RLs at concentrations ranged from 5 to 50 mg L-1) showed significantly decreased levels of growth rate, while increased levels of Chl a and b were obtained only in 72-96 h RLs-treated algae. Similarly, although no changes were obtained in the Chl a/b ratio and almost all chlorophyll fluorescence parameters over time, yields of electron transport (ϕR0, ϕE0) and respective performance index (PItotal) were negatively affected at 72 and 96 h. Based on those findings, it seems that the inhibitory effect of RLs on the algae growth rate after 24 and 48 h and the gradual attenuation of the phenomenon (after 72 h of exposure), may indicate the initial response of the organism, as well as algae ability to overcome, since RLs showed no effects on algae photosynthetic ability. Those findings reveal for the first time that RLs from Burkholderia thailandensis are not harmful for Dunaliella tertiolecta. However, further studies with the use of more aquatic species could be essential for assessing the RLs-mediated effects on aquatic biota.

Treatment of printing ink wastewater using electrocoagulation.

The present study investigates the treatment of real printing ink wastewater by using the electrocoagulation (EC) process. Effects of initial chemical oxygen demand (COD) concentrations, electrode materials and current densities were examined to determine the maximum COD and color removal from the wastewater. In parallel, raw and treated printing ink wastewater toxic potential was further estimated via the application of toxicity tests using the freshwater crustacean Thamnocephalus platyurus for assessing EC process efficiency. According to the results, it was observed that the EC is efficient under most of the operating conditions used, as COD and color removal ranged between 72.03 to 85.81% and 98.7-100%, respectively. The total cost of the EC process, considering the treatment time, applied current, applied voltage and the total anode electrode mass consumption was also estimated. The Fe electrode proved to be of lower cost than the Al electrode, however the use of Al electrode produced better decolorization results in the solutions. Moreover, toxicity tests currently performed with the use of larvae of the fairy shrimp Thamnocephalus platyurus revealed a substantial decrease in the toxic potential of printing ink wastewater, thus indicating the efficiency of the proposed EC process.

Assessment of the toxic potential of rainwater precipitation: First evidence from a case study in three Greek cities.

Considering the necessity to monitor rainwater quality and the limited existing data regarding wet precipitations' toxic potential, the toxic potential of rainwater (RW) samples collected in three cities of Western Greece (Corfu, Agrinion and Patras) during May, October, November, and December of 2014 were investigated. The relationship of the adverse effects obtained, against cytokinesis block micronucleus (CBMN) and Vibrio fischeri bioluminescence (Microtox) assays with the physicochemical characteristics, the heavy metal concentration levels of rainwater samples and the meteorological parameters governing the climatic conditions in each region, were studied in detail. All samples showed pH values in the range of 5.37-7.17, high concentrations of ΝΗ4+ and the presence of SO42- and NO3-. The metals Al, Mn, Fe, Co, Ni, Cu, Zn, As, Sb, Cd, were detected in most of the samples, with maximum concentrations ranging from 0.16 µg L-1 (for Co) to 86.6 µg L-1 (for Fe). The CBMN assay showed statistically significant rainwater cytotoxic activity at 5% (v/v) concentration in all cities during all months studied and at 10% (v/v) in Agrinion and Corfu in May. No genotoxic effect was induced at 5% (v/v) concentration of rainwater, except for Corfu city during November, whereas 10% (v/v) of rainwater led to statistically significant increase in micronuclei frequencies in all three cities. The highest (100%) and the lowest (14%) inhibitory effect exhibited by the rainwater in the Vibrio test were noticed at Corfu city during May and December, respectively. Considering the inter-relations obtained among the abiotic and biotic variables measured in all cases, conductivity, SO42-, NH4+, and total metal burden could be related with both CBMN and Microtox assays data, regardless of the site- and month-specific differences. The multidisciplinary approach presented here showed that the physicochemical profile of rainwater precipitation could be significantly related with the induction of adverse effects on biota.