Genotoxicity screening of the river Rasina in Serbia using the Allium anaphase–telophase test

Evaluation of the presence of genotoxic substances is especially important in rivers that serve as a source of drinking water. Nine water samples collected along the river Rasina in Serbia were analyzed for potential toxic and genotoxic effects using the Allium anaphase-telophase test. Inhibition of root growth relative to the negative control (synthetic water) was observed in all samples. Analysis of the genotoxic potential, through scoring anaphase and telophase aberrations, showed that in seven of the nine samples the level of aberrations was significantly increased relative to the negative control but was lower than that obtained for the positive control (methyl methanesulfonate). Changes in the relation between spindle and chromosome types of aberrations were found in some samples, indicating differences in the potential genotoxic substances present at the analyzed sites. The data, which were obtained from samples collected at the highest level of river water, warn that during periods of low flow the values could reach genotoxic activity. The Allium anaphase-telophase test can be recommended as an monitoring system, that can serve as the first alert for the presence of genotoxic environmental pollutants.     

The chemical and biogenotoxic characterization of organic xenobiotics in aquatic sediment materials 1. The application and comparison of chemically non-specific and biogenotoxic methods

The aim of this work was to evaluate the Ames assay and mixed function oxidase (MFO)-Induct Test used in parallel with chemical group tests (ECD fingerprint and PAH estimation) for the characterization of the organic pollution of water sediment materials. Sediment materials were collected from “clean” and relatively heavily polluted locations in the Middle Adriatic Sea, and from some locations in continental Croatia polluted with wastewaters from different enterprises. Characterization of the organic extracts of the sediment materials investigated was performed chemically using UV spectrofluorometry for the determination polyaromatic hydrocarbons (PAH) and gas chromatography for the determination of volatile EC detector sensitive materials. Genotoxic analysis of the extracts was performed using the MFO-Induct Test and mutagenicity testing using the Standard Plate Incorporation Test as described by Maron and Ames with Salmonella typhimurium TA 98. Measurement of the BaPMO enzyme activity in the livers of carp treated i.p. with total extracts of the sediment investigated confirmed that the methanol extracts generally contained more inducing matter than the petroleum ether extracts. Ames assay showed that for all the samples following the elimination of the sulfur, there was an increase in the number of revertants in comparison to the control number, which indicates that the samples contained mutagenic substances. The larger doses of extracts generally demonstrated cytotoxicity, as evidenced by a reduced number of spontaneous revertants in the Salmonella/Microsome Test. Investigation of the correlation of the chemical parameters with the biological parameter showed that the induction of BaPMO exhibited a statistically significant correlation with the level of the ECD fingerprint of the petroleum ether sediment extract.     

Determination of selected neonicotinoid insecticides by liquid chromatography with thermal lens spectrometric detection

We studied the analysis of trace amounts of neonicotinoid insecticides by liquid chromatography coupled with a thermal lens spectrometric detector (TLS). This multi-residue analysis method is based on the reversed phase separation on C₁₈ column, isocratic elution and collinear dual beam TLS detection. The insecticides thiamethoxam, imidacloprid, acetamiprid and thiacloprid were detected with retention times of 4.4, 5.7, 6.5 and 8.5 min and limits of quantifications of 50, 89, 10, and 25 μg/L, respectively. The retention times agreed well with those obtained by the same chromatographic method but using a diode-array detector (DAD). The limits of quantifications for imidacloprid were identical in both techniques. However, the limits of quantifications for thiamethoxam, acetamiprid and thiacloprid were up to 8.5 times lower using the TLS detector compared to the diode-array detector. The applicability of the developed procedure was tested on spiked river water and potato samples.     

The Role of Suspended Matter in Assessing the Assimilative Capacity Case Study of Two Estuaries in the Adriatic Sea

Suspended particulate matter has been studied in two Adriatic river mouths to assess its role as a transport vehicle for waste disposal into the Adriatic sea.

The larger of the two, the Adige River (northwestern Adriatic coast, Italy), carries large quantities of solids and builds up a prodelta. the Krka River (eastern Adriatic coast, Croatia, Yugoslavia) is a typical karst river, and carries small quantities of suspended matter, since it drains carbonate terrains.

The resulting disposal of contaminants into the Adriatic sea is largely governed by the adsorption/desorption processes occurring in the estuarine mixing region. Surface adsorption properties, the specific surface area, and the nature and reactivity of the organic film coating determine the partitioning of pollutants between the dissolved and particulate state.

The observations reported point to the key role of particulates in transport of pollutants from land to sea. the concept of the assimilative capacity of the estuarine area requires detailed scientific knowledge on the geochemical behaviour of particulates.

In the Adige River mouth, lead, chromium, zinc, and nickel are accumulated in the prodelta sediments, and thus the loading of the sea with these metals is diminished, whereas in the Krka River estuary, desorption and mobilization of some trace metals occurs.     

LCA of tomato greenhouse production using spatially differentiated life cycle impact assessment indicators: an Albanian case study

The increasing attention to agricultural exports and sustainability issues is driving a surge of interest in the life cycle assessment (LCAs) of greenhouse crop production in Albania. Meanwhile, most of the reported agricultural LCAs tend to be generic without considering regionalized environmental sensitivities. In this study, ReCiPe 2016, covering 18 midpoint indicators and 3 endpoint indicators was used to generate a full-fledged cradle-to-farm gate LCA of greenhouse tomatoes in a typical Albanian farm including spatial differentiation and indicators not covered by contemporary LCAs. The most important midpoint categories per 1 ha identified from foreground–background analysis were global warming (2660.4 kg CO2-eq), stratospheric ozone depletion (0.0308 kg CFC11-eq), particulate matter formation (7.99 kg PM2.5-eq), human health and ecosystem ozone formation (8.47 and 14.95 kg NOx-eq), water consumption (2293.23 m3), and terrestrial acidification (42.28 kg SO2-eq). The application of spatial differentiation resulted in higher impacts with about 21% for particulate matter formation, 12% for human health ozone formation, 134% for ecosystem ozone formation, 19% for terrestrial acidification, and 13% for water consumption. The impacts primarily originated from nitrogen-based fertilizer emissions and diesel fuel with the origin of the impact from nitrous oxide (N2O), ammonia volatilization (NH3), nitrogen oxides (NOx), and non-methane volatile organic compounds (NMVOCs). Water consumption was dominated by irrigation water use. Overall, at the endpoint level, 9% and 24% less cumulative damage to human health and ecosystem quality were calculated with respect to the site-generic analysis primarily from the cause-and-effect chain of water consumption (mainly lower water stress index). This affirms the importance of regional considerations in LCA calculations to reflect the impacts accordingly (i.e., the magnitude of impacts, the most relevant midpoint categories, and their relevance on endpoint level) and increase the possibility of making correct conclusions and sub-optimizations, i.e., increase the discriminating power of LCA.     

Investigation of the production of lipophilic volatile organohalogen materials during laboratory chlorination of some ground water samples from Yugoslavia

Water chlorination leads to the generation of various halogenated products of natural and waste water organic matter. In natural surface and ground water humic matter usually makes the greatest part of the total organic matter. However, in the water polluted with various organic wastes, other organic matter is also present in significant concentrations. The investigation of the relationship between the concentration of organic matter in the samples of ground water in Yugoslavia on one side, and the production of the lipophilic volatile organohalogen materials during the process of their chlorination on the other side is described. The production of the halomethanes and other ECD response materials during the chlorination of water samples in laboratory conditions is compared with the concentrations of organic materials in water samples. From the obtained linear correlation coefficients, possibilities of predicting amounts of organohalides during the chlorination process by using several very simple methods for the organic materials estimation in the investigated water samples are discussed and compared.     

Minimization of blank values of the miniature silica gel column

Several methods involving silica gel separations of polychlorinated biphenyls from chlorinated insecticides are already described in literature. Our intention was to find a system of solvent and silica gel of the lowest blank values. The first step was to find a solvent mixture which will keep the homogenity of a silica gel column. PCBs elution was performed with pentane, while the chlorinated insecticides eluated with diethylether, acetone, methanol, and benzene. The elution of pesticides gave the best results for the following solvent systems: diethylether + hexane (1:1), acetone + hexane (1:4), methanole + pentane (1:8) and benzene.Separation of pesticides from polychlorinated biphenyls found in a synthetic mixture was also examined. It was found that all solvent systems used gave similar results of separation. However, only if benzene is used as eluant, the silica gel column can be used for four successive separations after the “on column” regeneration. In order to obtain the lowest blank values silica gel was cleaned with several solvents. The eluates after the silica gel separation were also cleaned up by means of deactivation alumina and concentrated sulphuric acid.     

Oxidation of organophosphorus pesticides with chloroperoxidase enzyme in the presence of an ionic liquid as co-solvent

Effects of selected ionic liquids on the efficiency of CPO oxidation of methyl-parathion were studied. In general, the activity of CPO decreases with the increased concentration of ionic liquid. For ionic liquids with the same cation, those with PF₆ ^? exhibit strongest inhibition toward CPO, while those with either BF₄ ^? or metSO₄ ^? show relatively lesser inhibition. It was further demonstrated that even for buffer solutions containing up to 30% concentration of ionic liquids such as EAN, BMIMmetSO₄, and EtPyTFA, CPO still can retain high activity (70–100% compared to that in citrate buffer) for oxidation of methyl-parathion. Photothermal enhancement factors of 3.5 times and corresponding improvements of sensitivity in the determination of organophosphates by the FIA-TLS method are predicted in 30% ionic liquids.     

Photolytic and photocatalytic degradation of 6-chloronicotinic acid

This work describes for the first time the photolytic and photocatalytic degradation of 6-chloronicotinic acid (6CNA) in double deionised water, which is a degradation product of neonicotinoid insecticides imidacloprid and acetamiprid, and it is known to appear in different environmental matrices. Photolytic experiments were performed with three UVA (ultraviolet A) polychromatic fluorescent lamps with broad maximum at 355 nm, while photocatalytic experiments were performed using immobilised titanium dioxide (TiO₂) on six glass slides in the spinning basket inside a photocatalytic quartz cell under similar irradiation conditions. Photolytic degradation revealed no change in concentration of 6CNA within 120 min of irradiation, while the photocatalytic degradation within 120 min, obeyed first-order kinetics. The observed disappearance rate constant was k = 0.011 ± 0.001 min⁻¹ and t_1/2 was 63.1 ± 5.5 min. Mineralisation rate was estimated through total organic carbon (TOC) and measurements revealed no carbon removal in case of photolysis after 120 min of exposure. However in photocatalytic experiments 46 ± 7% mineralisation was achieved within 120 min of irradiation. Nevertheless, the removal of total nitrogen (TN) was not observed across all experiments. Ion chromatographic analyses indicated transformation of chlorine atoms to chloride and increase of nitrate(V) ions only via photocatalytic experiments. Efficiency of selected advanced oxidation process (AOP) was investigated through toxicity assessment with Vibrio fischeri luminescent bacteria and revealed higher adverse effects of treated samples on bacteria following photocatalytic degradation in spite of the fact that higher mineralisation was achieved. New hydroxylated product generated in photocatalytic experiments with TiO₂, was confirmed with liquid chromatography-electro spray ionisation mass spectrometry (LC-ESI-MS/MS) analyses, gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (¹H NMR).     

Evaluation of modifications of the simple spectrofluorometry method for estimating petroleum hydrocarbon levels in fresh and waste water samples

The monitoring of petroleum hydrocarbons in the water environment is very often performed using complicated methods. There are many difficulties encountered in analyzing a long number of samples in the monitoring programme. In such cases, a simpler though less precise and specific technique using inexpensive equipment to monitor a large number of water samples, might be more desirable.

This idea was realized in the assessment of the state of pollution of the Mediterranean Sea by petroleum hydrocarbons where the pollution of sea water with oil (dissolved and/or dispersed in sea water) was measured by spectrofluorometry. The paper describes an analytical evaluation of three modifications of simple the spectrofluorometry method, applied on numerous river and waste water samples collected from various areas of Croatia.