Heavy metal contamination of paddy soils and rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) from Kočani Field (Macedonia)

This research focuses on the heavy metal contamination of the paddy soils and rice from Kočani Field (eastern Macedonia) resulting from irrigation by riverine water impacted by past and present base-metal mining activities and acid mine drainage. Very high concentrations of As, Cd, Cu, Pb and Zn were found in the paddy soils (47.6, 6.4, 99, 983 and 1,245 μg g⁻¹) and the rice (0.53, 0.31, 5.8, 0.5 and 67 μg g⁻¹) in the western part of Kočani Field, close to the Zletovska River, which drains the mining facilities of the Pb–Zn mine in Zletovo. In terms of health risk, the observed highest concentrations of these elements in the rice could have an effect on human health and should be the subject of further investigations.     

Element levels in cultured and wild sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) from the Adriatic Sea and potential risk assessment

In this study, the role of aquaculture activity as a source of selected metals was analyzed. Significant differences in element content between cultured (Dicentrarchus labrax, Sparus aurata) and wild fishes as well as between fish muscle and their feed were detected. Higher concentrations of trace elements (i.e., As, Cu, Hg, Se) in wild fish tissues in comparison with cultured ones indicate additional sources of metals beside fish feed as natural and/or anthropogenic sources. Generally, mean Cd, Cu, Pb, Se, and Zn concentrations in cultured (0.016, 1.79, 0.14, 0.87, and 34.32 μg/g, respectively) and wild (0.011, 1.97, 0.10, 1.78, and 23,54 μg/g, respectively) fish samples were below the permissible levels, while mean As (2.57 μg/g in cultured, 4.77 μg/g in wild) and Cr (5.25 μg/g in cultured, 2.92 μg/g in wild) values exceeded those limits. Hg values were lower in cultured (0.17 μg/g) and higher in wild (1.04 μg/g) fish specimens. The highest elemental concentrations were observed in almost all fish samples from Kor?ula sampling site. The smallest cultured sea basses showed As (4.01 μg/g), Cr (49.10 μg/g), Pb (0.65 μg/g), and Zn (136 μg/g) concentrations above the recommended limits; however, values decreased as fish size increased. Therefore, the majority of metal concentrations in commercial fishes showed no problems for human consumption. Also calculated Se:Hg molar ratios (all >1) and selenium health benefit values (Se-HBVs) (all positive) showed that consumption of all observed fishes in human nutrition is not risk.     

Terrestrial Biodiversity Analyses in Dalmatia (Croatia): A Complementary Approach Using Diversity and Rarity

Here we present the methodology used for terrestrial biodiversity analysis and site selection in Phase B of the UNDP/GEF COAST project. The analysis was focused on the problem of biodiversity evaluation in four Croatian counties stretching from sea level to the highest mountain in Croatia. Data on habitats, vascular flora, and fauna (mammals, birds, reptiles, amphibians, butterflies, ground beetles, and underground invertebrates) were collected and analyzed for each of the four counties. Emphasis was given to the richness of endangered species and the rarity of endemic species. Based on the spatial analyses of habitat, fauna, and flora data, four to six areas were selected from each county and ranked according to their biodiversity importance. Overlap between areas important for richness and those important for rarity was highest for data on flora (65.5%) and lowest for data on fauna (16.7%). When different data sets were compared, the lowest overlap was between flora and fauna (17.1%) and largest between fauna and habitats (23.9%). Simultaneous overlap among all three data sets was found in just 6.5% of the overall selected areas. These results suggest that less specific data, with respect to taxa threat status, could better serve as surrogate data in estimating overall biodiversity. In summary, this analysis has demonstrated that Dalmatia is a region with a high overall biodiversity that is important in a broader European context.     

Major, minor and trace element content derived from aquacultural activity of marine sediments (Central Adriatic, Croatia)

Introduction and purpose

Studies examining the environmental impact of marine aquaculture have increased significantly in number during the last few decades. The present paper investigates a region of rapid growth in intensive aquaculture and its influence on the local marine ecosystem.

Discussion

This study was undertaken with the specific aim of assessing the effect of fish farming on marine sediment at a farm near the island of Vrgada in the Central Adriatic. Data obtained regarding major (Si, Al, K, Na, Fe, Ca, Mg), minor (Mn, P, Ti) and trace (As, Au, Ba, Cd, Co, Cr, Cs, Cu, Ga, Hf, Hg, Mo, Nb, Ni, Pb, Rb, Sb, Sc, Se, Sr, Ta, Th, Tl, U, V, Y, Zn, Zr) elements were used to estimate the spatial and temporal distribution of metals in the sediment and their possible relationship with local aquacultural activity.

Results

Although the measured concentrations of heavy metals in sediment below fish cages were notably different and potentially a result of farming activity, the values were generally lower than background concentrations observed in the Central Adriatic. In contrast, concentrations of heavy metals at a reference site unaffected by aquaculture varied from lower levels to values even higher than those observed below the high-production cages. Furthermore, calculated environmental index values indicate that the sediment below the farm is either uncontaminated or suffers from only low levels of contamination.

Conclusion

Such results suggest that the effect of observed fish farm activity on the local marine ecosystem is practically negligible.     

Use of sensitive methods for detection of DNA damage on human lymphocytes exposed to p,p′-DDT: Comet assay and new criteria for scoring micronucleus test

Wide distribution, stability and long persistence in the environment of dichlorodiphenyltrichloroethane (DDT), probably the best-known and most useful insecticide in the world, imposes the need for further examination of the effect of this chemical on human health and especially on the human genome. In this study, peripheral blood human lymphocytes from a healthy donor were exposed to 0.025 mg/L concentration of p,p′-DDT at different time periods (1, 2, 24 and 48 h). For the assessment of genotoxic effect, the new criteria for scoring micronucleus test and alkaline comet assay were used. Both methods showed that p,p′-DDT induces DNA damage in low concentration used in this research. Results of micronucleus test showed a statistically significant (p < 0.05) genotoxic effect of p,p′-DDT on human lymphocytes compared with corresponding control and a different exposure time. A comet assay also showed increased DNA damage caused in p,p′-DDT-exposed human lymphocytes than in corresponding control cells for the tail length. Results obtained by measuring the level of DNA migration and incidence of micronuclei (MN), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) indicate the sensitivity of these tests and their application in detection of primary genome damage after long-term exposure to establish the effect of p,p′-DDT on human genome.     

Toxicity of mancozeb,chlorothalonil, captan,fluopyram, boscalid,and difenoconazole to Didymella applanata isolates from Serbia

Field isolates of Didymella applanata, the causal agent of spur blight of raspberry, were evaluated in vitro for their sensitivity to mancozeb, chlorothalonil, captan, fluopyram, boscalid and difenoconazole. A total of 10 isolates, collected during 2013 at five localities in the major raspberry growing region in Serbia, and characterized as copper hydroxide, dithianon, and tebuconazole (sensitive), pyraclostrobin (sensitive or highly resistant) and fluazinam (sensitive or moderately resistant), were used in this study. The EC₅₀ values for the isolates ranged from 1.33 to 2.88 mg L^?1 for mancozeb, from 3.18 to 6.65 mg L^?1 for chlorothalonil, from 15.75 to 24.69 mg L^?1 for captan and from 1.80 to 8.20 mg L^?1 for fluopyram. The narrowest range of EC₅₀ values was recorded for difenoconazole (0.23–0.49 mg L^?1), whereas the widest range was obtained for boscalid (4.49–49.25 mg L^?1). The calculated resistance factors showed that all D. applanata isolates were sensitive to mancozeb, chlorothalonil, captan, and difenoconazole. Four isolates were moderately resistant to boscalid, while three of them were also moderately resistant to fluopyram. This finding of moderately resistant isolates to these SDHI fungicides indicates a possible cross-resistance which should be clarified in further investigations.     

Genotoxic potential of selected cytostatic drugs in human and zebrafish cells

Due to their increasing use, the residues of anti-neoplastic drugs have become emerging pollutants in aquatic environments. Most of them directly or indirectly interfere with the cell’s genome, which classifies them into a group of particularly dangerous compounds. The aim of the present study was to conduct a comparative in vitro toxicological characterisation of three commonly used cytostatics with different mechanisms of action (5-fluorouracil [5-FU], cisplatin [CDDP] and etoposide [ET]) towards zebrafish liver (ZFL) cell line, human hepatoma (HepG2) cells and human peripheral blood lymphocytes (HPBLs). Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and acridine orange/ethidium bromide staining. All three drugs induced time- and dose-dependent decreases in cell viability. The sensitivity of ZFL and HepG2 cells towards the cytotoxicity of 5-FU was comparable (half maximal inhibitory concentration (IC₅₀) 5.3 to 10.4 μg/mL). ZFL cells were more sensitive towards ET- (IC₅₀ 0.4 μg/mL) and HepG2 towards CDDP- (IC₅₀ 1.4 μg/mL) induced cytotoxicity. Genotoxicity was determined by comet assay and cytokinesis block micronucleus (CBMN) assay. ZFL cells were the most sensitive, and HPBLs were the least sensitive. In ZFL cells, induction of DNA strand breaks was a more sensitive genotoxicity endpoint than micronuclei (MNi) induction; the lowest effective concentration (LOEC) for DNA strand break induction was 0.001 μg/mL for ET, 0.01 μg/mL for 5-FU and 0.1 μg/mL for CDDP. In HepG2 cells, MNi induction was a more sensitive genotoxicity endpoint. The LOEC values were 0.01 μg/mL for ET, 0.1 μg/mL for 5-FU and 1 μg/mL for CDDP. The higher sensitivity of ZFL cells to cytostatic drugs raises the question of the impact of such compounds in aquatic ecosystem. Since little is known on the effect of such drugs on aquatic organisms, our results demonstrate that ZFL cells provide a relevant and sensitive tool to screen genotoxic potential of environmental pollutant in the frame of hazard assessment.     

Alteration of cholinesterase activity as possible mechanism of silver nanoparticle toxicity

Due to their broad-spectrum antimicrobial activity, silver nanoparticles (AgNPs) have been used in a large number of commercial and medical products. Such proliferated AgNP production poses toxicological and environmental issues which need to be addressed. The present study aimed to investigate the effects of AgNPs on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), important enzymes in areas of neurobiology, toxicology and pharmacology. Three different AgNPs, prepared by the chemical reduction using trisodium citrate, hydroxylamine hydrochloride (Cl-AgNPs), and borohydride following stabilization with poly(vinyl alcohol), were purified and characterised with respect to their sizes, shapes and optical properties. Their inhibition potential on AChE and BChE was evaluated in vitro using an enzyme assay with o-nitrophenyl acetate or o-nitrophenyl butyrate as substrates, respectively. All three studied AgNPs were reversible inhibitors of ChEs. Among tested nanoparticles, Cl-AgNP was found to be the most potent inhibitor of both AChE and BChE. Although the detailed mechanism by which the AgNPs inhibit esterase activities remains unknown, structural perturbation of the enzyme may be the common mode of ChE inhibition by AgNPs.