M. Petala

Influence of ozonation on the in vitro mutagenic and toxic potential of secondary effluents

Reclamation of municipal effluents by advanced treatment processes is an attractive perspective for facing certain water shortage problems. However, the application of tertiary techniques should be thoroughly examined for their potential hazardous effects. Ozonation is an efficient chemical oxidation method, often used in wastewater reclamation, which may result in by-products that may alter the toxic and mutagenic properties of effluents. In this study, Ames test and Microtox test were used for the evaluation of ozonation efficiency to upgrade secondary effluents quality. In general, the toxic response and mutagenic effect without metabolic activation of test species were influenced mainly by the ozone dose and ozonation duration, whereas the mutagenic effect with metabolic activation was influenced mainly by ozone dose, indicating that ozone conditions strongly affect the formation of by-products. In most cases, the toxicity was increased and reached up to 100% (in relation to that of secondary effluent) after ozonation with 8.0 mg O3/L for 5 min. On the contrary, in most cases the mutagenic activity towards strain TA98 without metabolic activation was reduced, when ozone dose and contact time increased. However, the mutagenicity was also increased after ozonation at low ozone doses and for contact times less than 5 min. The mutagenic activity of treated effluents towards strain TA98 with metabolic activation remained about the same or was reduced, compared to that of secondary effluent, and was even eliminated after ozonation with 8.0 mg O3/L for contact times higher than 5 min.

Bioavailability and Toxicity of Heavy Metals in the Presence of Natural Organic Matter

The effect of humic acids (HA) on the toxicity of copper, zinc and lead was investigated in this study, using the photobacterium Vibrio fischeri (Microtox test) as a test organism. The metal species and the complexation capacity of HA were predicted using model WinHumicV for speciation modeling. The toxicities of copper and lead were generally comparable, while the toxicity of zinc was lower than the ones of the other two method. The toxicity of copper decreased with the addition of HA, while the toxicity of zinc remained almost constant. The complexation of Zn with HA was low. Thus the presence of HA did not significantly influence the toxicity of Zn. For Cu and Zn, the free metal ion species showed to be basically responsible for the toxicity of these metals, while for Pb the chlorocomplexes and Pb-HA complexes contributed to the toxicity of Pb, indicating that the toxicity of heavy metals depends not only on the free metal ion species concentration but on different metal species that may be bioavailable.

Environmental hazard assessment of coal fly ashes using leaching and ecotoxicity tests

The environmental hazard of six coal fly ash samples collected from various coal incineration plants were examined in the present study, using various leaching tests along with physicochemical and ecotoxicological analyses. Physicochemical analyses indicated that the concentration of Ba, Cr, Mo, Se, sulfates and total dissolved solids (TDS) exceeded the threshold values, according to the European Council decision 2003/33/EC for inert, non-hazardous or hazardous waste. The highest sensitivity, among examined organisms, was presented by Daphnia magna and was ascribed to the lowest tolerance of the organism to Cr. The toxic effect profile of the leachates toward Vibrio fischeri revealed that its bioluminescence was significantly altered by the presence of heavy metals in the leachates (such as Cu, Ni and Zn) in low or sub-lethal doses. Although the toxicity of the leachates toward Pseudokirchneriella subcapitata was relatively high, the correlation between the toxicity and heavy metals concentration was rather poor, indicating the significance of the bioavailability of constituents and the importance of toxicity testing. Based on the classification of the samples resulted by physicochemical analyses of the leachates and the results of ecotoxicity tests, Physicochemical and Ecotoxicological indices were also defined, for an overall evaluation of the environmental impact assessment of fly ashes. The suggested indices showed that ecotoxicity tests were in some cases inconsistent with the outcome of waste classification based on physicochemical analyses. Moreover, the examination of leaching protocols and obtained results showed that one stage leaching test (L/S=10 L/kg) underestimated the fly ash classification, when compared with the two stage test EN 12457-3 at the cumulative ratio L/S=10 L/kg. In addition, the application of availability test NEN 7341 provided useful results about the ecotoxicity wastes. Thus, NEN 7341 is strongly suggested for the implementation of such studies, since the bioavailability of constituents may not be altered by pH adjustment tactics and pH-originated toxicity effects may be avoided.

Toxicological and ecotoxic impact of secondary and tertiary treated sewage effluents.

Secondary sewage effluents are discharged in significant quantities in aquatic environments delivering pollutants that were not removed during treatment; yet advanced treated effluents are not lacking of contaminants. In this study, biochemical biomarkers were measured in liver and kidney of rainbow trout (Oncorynchus mykiss) exposed to unchlorinated, chlorinated and tertiary treated secondary sewage effluents. In addition, organic matter, nitrogen and suspended solids were assayed, while a common bioassay, Daphnia magna 21d reproduction test was also applied in order to examine potential relation between the performed bioassay and the biomarkers. Processes using oxidative conditions, such as ozonation and chlorination, resulted in significantly increased breeding rate (up to 74%) of the organism. Biomarkers measurements incorporated the determination of total glutathione (GSH), glutathione S-transferases (GST), glutathione peroxidase (GPX), lipid peroxidation (LPO) and an innovative biomarker in such applications, haem peroxidase. In general, the response of biomarkers was dependent upon the treatment method and it was tissue specific. Secondary effluents inhibited liver GST and haem peroxidase, while GSH levels and LPO were significantly provoked in liver. Ozonation provoked hepatic peroxidation, in terms of haem peroxidase and LPO, and GST; while the protective (to Reactive Oxidant Species - ROS) GSH was depleted, suggesting extended ROS attack to the organism. Similar response of biomarkers (but to a lesser extend) was observed after exposure of trout to effluents submitted to both coagulation and ozonation, emphasizing the significance of removing the residual organic matter by other methods than oxidative ones. Ozonation also enhanced renal LPO and GPX; however the former employment of coagulation limited the peroxidation phenomena. Chlorination mainly affected the levels of total GSH in both tissues.

Potential Ozone Applications for Water/Wastewater Treatment

Abstract Several applications of ozonation were examined in this study for: i. the treatment of stabilized high strength municipal landfill leachates, ii. the reclamation potential and toxicity reduction of municipal secondary effluents, and iii. the removal potential of phytoplanktons from surface waters. The major parameters examined were the applied ozone dosage and the respective contact time. The application of single ozonation on leachates resulted in the efficient removal of color and organic loading, due to the respective oxidation, induced by ozonation. In addition, ozonation was found to be effective for the removal of the residual organic content of secondary municipal effluents. However, acute toxic effects after ozonation were observed on V. fischeri and were related to ozone concentration and contact time. Furthermore, the surface water used for drinking water production, was subjected to ozonation treatment for the removal of harmful cyanobacteria. Ozonation resulted in the reduction of the number of cyanobacteria species and in the breakage of the chain‐type species to cells with a lower number of atoms.

Hexavalent chromium release from lignite fly ash and related ecotoxic effects

Hexavalent chromium Cr(VI) is a pollutant of immense concern due to its high mobility to water sources and highly toxic properties. In most cases, Cr(VI) could be released from lignite fly ash in aquatic environment when fly ash comes into contact with water. In this study, the contribution of the leaching patterns and bioavailability of Cr(VI) from lignite fly ash to the overall ecotoxic properties of fly ash leachates was originally examined and leaching procedures were evaluated in this context. A series of customized leaching tests were conducted and a battery of ecotoxicity tests including the crustacean Daphnia magna and the photobacterium Vibrio fischeri was applied. The leaching of Cr(VI) was pH and liquid to solid (L/S) ratio dependent, exhibiting the highest releases at pH values between 7 and 8. At the liquid to solid ratio (L/S) equal to 100 L/kg, the (CrVI) release reached a plateau, implying the presence of diffusion constrains and/or solubility hindrances. The toxic effect of the leachates obtained under leaching at pH 7 towards D. magna was relatively high (TU = 28.6 (23.8–35.7) at L/S = 10 L/kg). Interestingly, the toxicity of the leachates towards D. magna not only was significantly correlated to Cr(VI) (r = 0.961, P < 0.01), but the toxicity of the leachates (in absolute values) was matching the toxicity of the Cr(VI) revealing its remarkable contribution to the overall effect. In addition, the lower sensitivity of the bacteria V. fischeri when exposed to the leachates, along with the time dependence of the toxicity profiles supported the interpretation of the results obtained in this study.

Comparison of several toxicity tests applied to complex wastewaters and mussel biomarkers in receiving waters

In this study, the complex wastewaters/effluents discharged to coastal regions of Turkey and Greece were sampled, and various toxicity tests were applied. The bioassays used included the assessment of the luminescence inhibition of the bacterium Vibrio fisheri, the photosynthesis inhibition (14C uptake rate) and growth inhibition of the algal species Phaeodactylum tricornutum, and the mortality of the crustacean Artemia franciscana and rotifer Brachionus plicatilis. Additionally, “neutral red retention” and “filtration rate” sublethal biomarker techniques were performed by utilizing the mussel species Mytilus galloproviancialis, inhabiting the points of discharges in Turkey. All discharges tested were found to be acutely toxic to P. tricornutum and slightly toxic to V. fisheri, A. franciscana, and B. plicatilis. Test results showed that the 14C uptake rate toxicity test was the most sensitive one among the other bioassays. The biomarker results showed that the health status of mussels in the coastal areas decreased significantly, indicating the pollution of the receiving waters in Turkey. The conclusions drawn highlight the necessity for an intesive ecotoxicological monitoring scheme that will incorporate the most suitable bioassays and biomarkers to adequately contribute to the upgrading and maintenance of the ecological quality of the coastal waters in Greece and in Turkey. *Presented as an invited plenary lecture at “The 2nd Asian International Conference on Ecotoxicology and Environmental Safety” (SECOTOX 2004), Songkla, Thailand, 26–29 September 2004.

Implications of handling practices on the ecotoxic profile of alumina nanoparticles towards the bacteria Vibrio fischeri.

ABSTRACT The complex nature and behavior of Engineered Nanomaterials (ENMs) has led to adoption of customized experimental ecotoxicity practices that are prone to possible artefacts in the inherent toxic properties of ENMs. In addition, the lack of standardized handling procedures for the ecotoxicity testing of ENMs prevents the development of experimental protocols for regulatory purposes. In this study, a suite of techniques for dispersion of ENMs was adopted and tested for two types of surface-modified alumina nanoparticles—one hydrophobic and one hydrophilic—towards the bacteria, Vibrio fischeri. The effect of certain handling practices on the observed ecotoxic effects on V. fischeri was examined. The overall goal was to evaluate by what means the handling practices of ENMs may affect the obtained toxicity results. It was realized that the toxicity of the hydrophilic and hydrophobic ENMs was mainly affected by the centrifugation and the salinity of the tested dispersions, respectively. It is more likely that both aluminium and coating substance contributed to the overall toxicity. Toxicity results are discussed with regard to generic physicochemical characteristics of the dispersions.