Monitoring of DNA damage in haemocytes of freshwater mussel Sinanodonta woodiana sampled from the Velika Morava River in Serbia with the comet assay

This study was undertaken to investigate the potential of the freshwater mussel Sinanodonta woodiana for detection of genotoxic pollution of the environment. Study was performed at two sites in the Velika Morava River, from May 2010 to February 2011. The alkaline comet assay on haemocytes was used, and the olive tail moment (OTM) was chosen as a measure of DNA damage. The specimens held on acclimation under controlled laboratory conditions for 10 d were used as a control. Chemical analysis revealed the presence of phosphates and increased concentrations of zinc, copper and nickel at both sites during the entire sampling period. The values of OTM in mussels collected from the environment, significantly correlated with the concentration of zinc (r = 0.6248), temperature (r = 0.7006) and dissolved oxygen (r = 0.7738). Seasonal variations in genotoxic response were observed, with the highest OTM values obtained during summer months. Preliminary results of the in vitro study indicated the effect of water temperature on genotoxic response to zinc and cadmium in S. woodiana suggesting that the presence of genotoxic pollutants during months with lower temperature could be under-estimated. Obtained results indicate that S. woodiana could be a valuable tool for active biomonitoring of aquatic environments and emphasizes the importance of seasonal genotoxic monitoring with this organism.     

Metal immobilization and phosphorus leaching after stabilization of pyrite ash contaminated soil by phosphate amendments

In this study we would like to show the importance of a holistic approach to evaluation of chemical stabilization using phosphate amendments. An extensive evaluation of metal stabilization in contaminated soil and an evaluation of the leaching of phosphorus induced after treatment were performed. The soil was highly contaminated with Cu (2894 mg kg(-1)), Zn (3884 mg kg(-1)), As (247 mg kg(-1)), Cd (12.6 mg kg(-1)) and Pb (3154 mg kg(-1)). To immobilize the metals, mixtures of soil with phosphate (from H(3)PO(4) and hydroxyapatite (HA) with varying ratios) were prepared with a constant Pb : P molar ratio of 1: 10. The acetic acid extractable concentration of Pb in the mixture with the highest amount of added phosphoric acid (n(H(3)PO(4)) : n(HA) = 3 : 1) was reduced to 1.9% (0.62 mg L(-1)) of the extractable Pb concentration in the untreated soil, but the content of water extractable phosphorus in the samples increased from 0.04 mg L(-1) in the untreated soil sample up to 14.3 mg L(-1) in the same n(H(3)PO(4)) : n(HA) = 3 : 1 mixture. The high increase in arsenic mobility was also observed after phosphate addition. The PBET test showed phosphate induced reduction in Pb bioavailability. In attempting to stabilize Pb in the soil with the minimum treatment-induced leaching of phosphorus, it was found that a mixture of soil with phosphate addition in the molar ratio of H(3)PO(4) : HA of 0.75 : 1 showed the most promising results, with an acetic acid extractable Pb concentration of 1.35 mg L(-1) and a water extractable phosphorus concentration of 1.76 mg L(-1). The time-dependent leaching characteristics of metals and phosphorus for this mixture were evaluated by a column experiment, where irrigation of the soil mixture with the average annual amount of precipitation in Slovenia (1000 mm) was simulated. The phosphorus concentration in the leachates decreased from 2.60 mg L(-1) at the beginning of irrigation to 1.00 mg L(-1) at the end.     

The impact of disturbance and ensuing forestry practices on Collembola in monitored stands of windthrown forest in the Tatra National Park (Slovakia)

Soil Collembola communities were investigated in spruce forest stands of the High Tatra Mts that had been heavily damaged by a windstorm in November 2004 and subsequently by a wildfire in July 2005. The study focused on the impact of these disturbances and forestry practices on collembolan community distribution and structure 4 years after the disturbance. Four different treatments were selected for this study: intact forest stands (REF), non-extracted windthrown stands (NEX), clear-cut windthrown stands (EXT) and burnt windthrown stands (FIR). From a total of 7,820 individuals, 72 species were identified. The highest total abundance mean was recorded in FIR stands followed by NEX and EXT stands and, surprisingly, the lowest in REF stands. The highest total species richness was observed in REF stands, followed by NEX stands and FIR stands and the lowest in EXT stands. In REF and NEX stands, the most abundant species were Folsomia penicula and Tetracanthella fjellbergi, while in heavily damaged stands, the most abundant was Anurophorus laricis. The ordination method used demonstrated a significant influence of treatment on the abundance of Collembola. ANOVA used confirmed significant differences for all dominant species between treatments. The present study shows the negative impact of windthrow on Collembola communities as reflected in decreased species richness and abundance. However, disturbance by fire caused a considerable increase in collembolan abundance 3 years after the event. Moreover, we show that clearing of windthrown spruce forests after a windstorm is less favourable for communities of soil collembolans and slows down the recovery process.     

Biomonitoring of heavy metals in fish from the Danube River

The Croatian part of the Danube River extends over 188 km and comprises 58 % of the country’s overall area used for commercial freshwater fishing. To date, the heavy metal contamination of fish in the Croatian part of the Danube has not been studied. The main purpose of this study was to determine heavy metal levels in muscle tissue of sampled fish species and to analyze the measured values according to feeding habits of particular groups. Lead ranged from 0.015 μg^?1 dry weight in planktivorous to 0.039 μg^?1 dry weight in herbivorous fish, cadmium from 0.013 μg^?1 dry weight in herbivorous to 0.018 μg^?1 dry weight in piscivorous fish, mercury from 0.191 μg^?1 dry weight in omnivorous to 0.441 μg^?1 dry weight in planktivorous fish and arsenic from 0.018 μg^?1 dry weight in planktivorous to 0.039 μg^?1 dry weight in omnivorous fish. Among the analyzed metals in muscle tissue of sampled fish, only mercury exceeded the maximal level (0.5 mg kg^?1) permitted according to the national and EU regulations determining maximum levels for certain contaminants in foodstuffs, indicating a hazard for consumers of fish from the Danube River.     

Application of artificial neural network in medical geochemistry

For the evaluation of various adverse health effects of chemical elements occurring in the environment on humans, the comparison and linking of geochemical data (chemical composition of groundwater, soils, and dusts) with data on health status of population (so-called health indicators) play a key role. Geochemical and health data are predominantly nonlinear, and the use of standard statistical methods can lead to wrong conclusions. For linking such data, we find appropriate the use method of artificial neural networks (ANNs) which enable to eliminate data inhomogeneity and also potential data errors. Through method of ANNs, we are able to determine the order of influence of chemical elements on health indicators as well as to define limit values for the influential elements at which the health status of population is the most favourable (i.e. the lowest mortality, the highest life expectancy). For determination of dependence between the groundwater contents of chemical elements and health indicators, we recommend to create 200 ANNs. In further calculations performed for identification of order of influence of chemical elements as well as definition of limit values, we propose to work with median or mean values from calculated 200 ANNs. The ANN represents an appropriate method to be used for environmental and health data analysis in medical geochemistry.     

Measurements of aerosol particles in the Škocjan Caves,Slovenia

For the first time, continuous aerosol measurements were performed in the ?kocjan Caves, one of the most important cave systems in the world, and listed by UNESCO as a natural and cultural world heritage site since 1986. Measurements of PM₁₀ were performed during three different periods: (1) in December 2011, the average background concentration was found to be about 4 μg m^?3; (2) in June 2012, a higher concentration was measured (8 μg m^?3); and (3) from 8 to 20 August 2012, the highest concentration of 15.3 μg m^?3 was measured. Based on the PM₁₀ measurement results, and as compared to similar measurements outside the cave, it can be hypothesized that the increase in the cave's aerosol concentration during the summer was connected to both the higher number of visitors and the polluted atmospheric air entering the cave upon entering of the cave system. Additional measurement of nanoparticles with scanning mobility particle sizer spectrometer (size between 14.1 and 710.5 nm) confirmed these findings; during the summer period, a severe raise in the total aerosol concentration of 30–50 times was found when groups of visitors entered the cave. Our results on nanoparticles demonstrated that we were able to detect very small changes and variations in aerosol concentration inside the cave. To our knowledge, these are the first results on nanoaerosol measurements in a cave, and we believe that such measurements may lead to the implementation of better protection of delicate cave systems.