The rhizosphere and PAH amendment mediate impacts on functional and structural bacterial diversity in sandy peat soil

To reveal the degradation capacity of bacteria in PAH polluted soil and rhizosphere we combined bacterial extradiol ring-cleavage dioxygenase and 16S rRNA analysis in Betula pubescens rhizoremediation. Characterisation of the functional bacterial community by RFLP revealed novel environmental dioxygenases, and their putative hosts were studied by 16S rRNA amplification. Plant rhizosphere and PAH amendment effects were detected by the RFLP/T-RFLP analysis. Functional species richness increased in the birch rhizosphere and PAH amendment impacted the compositional diversity of the dioxygenases and the structural 16S rRNA community. A shift from an Acidobacteria and Verrucomicrobia dominated to an Alpha- and Betaproteobacteria dominated community structure was detected in polluted soil. Clone sequence analysis indicated catabolic significance of Burkholderia in PAH polluted soil. These results advance our understanding of rhizoremediation and unveil the extent of uncharacterized functional bacteria to benefit bioremediation by facilitating the development of the molecular tool box to monitor bacterial populations in biodegradation.     

(210)Po and (210)Pb activity concentrations in Mytilus galloprovincialis from Croatian Adriatic coast with the related dose assessment to the coastal population

Activity concentrations of ²¹⁰Po and ²¹⁰Pb in the species Mytilus galloprovincialis from Croatian Adriatic coast were determined. The samples were collected in thirteen sampling places which are also a part of Croatian Mediterranean mussel watch project. The results of ²¹⁰Po and ²¹⁰Pb activity concentrations were found to vary between (22.1 ± 2.5)-(207 ± 21) Bq kg⁻¹ fresh weight and (2.8 ± 1.4)-(9.3 ± 0.7) Bq kg⁻¹ fresh weight, respectively. ²¹⁰Po/²¹⁰Pb ratio ranged between 6.2 and 30.7. The highest measured activities were found in the enclosed basins of the Neretva channel, where the majority of Croatian mussel cultivation is taking place. The estimated consequent average total annual effective ingestion dose due to ²¹⁰Po and ²¹⁰Pb from mussel consumption in Croatian coastal region is 202 ± 99 μSv with 96% contribution of ²¹⁰Po to the total effective dose.     

Impact of physical factors on the society and environment

In 1988 I published a paper “Electromagnetic fields—a new ecological factor” (Markov 1988). There was no internet and very few cell phones were available. It was not even fear from the hazard of the power lines. However, I want to cite the beginning of this paper: “The contemporary conditions of life put man in dependence of the complex of physical influences on the environment and in the first place—of the electromagnetic fields. The rapid development of science and technology has resulted in the introduction of many new devices and technologies in industry, agriculture, and everyday life. On the other hand, during their phylogenetic and ontogenic development, the living organisms are continuously exposed to the influence of different biotic and abiotic factors. The physical factors are included in the first group, and the entire evolution of life is connected with an adaptation to the action of these factors.” Could I repeat this statement nearly a quarter of century later? Sure, I will. The only difference would be that I will speak not only for electromagnetic fields, but for green-house effects, global warming, volcano eruption, and radiation disasters. I would also discuss the noise and vibration to which people are exposed from early days to the very end of their life traveling with private vehicles or public transport.     

Levels of ²¹⁰Po and ²¹⁰Pb in fish and molluscs in Slovenia and the related dose assessment to the population

²¹⁰Po and ²¹⁰Pb activity concentrations in fish from the Slovenian part of Adriatic Sea, in the vicinity of a former uranium mine at ?irovski vrh and from the Slovenian market were determined. In addition, ²¹⁰Po and ²¹⁰Pb activity concentrations in squid from the Slovenian market and in mussels from the Slovenian part of the Adriatic Sea were also determined. Fish, squid and mussel consumption in Slovenia was assessed from the data available from Eurostat and Food and Agriculture Organisation (FAO) and the data used for the corresponding dose calculation. Fish species with the highest activity concentrations were grilled to assess possible loss of ²¹⁰Po during the food preparation process. Samples were freeze dried and radiochemical separation of ²¹⁰Po and ²¹⁰Pb was performed. Measurements of ²¹⁰Po were performed by alpha spectrometry and ²¹⁰Pb by a low background gas-flow proportional counter. ²¹⁰Po activity concentrations in fish, squid and mussels were from 0.039 to 35.0 Bq kg⁻¹ fresh weight and ²¹⁰Pb activity concentrations were from 0.08 to 3.03 Bq kg⁻¹ fresh weight. Grilling of fish resulted in no significant loss of ²¹⁰Po at 90 °C. The assessed combined annual effective ingestion dose due to ²¹⁰Po and ²¹⁰Pb for fish, squid and mussels consumed in Slovenia is 47.6 μSv year⁻¹.     

Short- and long-term patterns of ¹³⁷Cs in fish and other aquatic organisms of small forest lakes in southern Finland since the Chernobyl accident

We summarize the patterns of ¹³⁷Cs activity concentrations and transfer into fish and other biota in four small forest lakes in southern Finland during a twenty-year period following the Chernobyl accident in April 1986. The results from summer 1986 showed fastest accumulation of ¹³⁷Cs into planktivorous fishes, i.e. along the shortest food chains. Since 1987, the highest annual mean values of ¹³⁷Cs have been recorded in fish occupying the highest trophic levels, for perch (Perca fluviatilis) 13,600 Bq/kg (ww) and for pike (Esox lucius) 20,700 Bq/kg (ww). At the same time, activity concentrations of ¹³⁷Cs in crustacean zooplankton and Asellus aquaticus have ranged between 1000 and 19,500 Bq/kg (dw). In 2006, 5-28% of the 1987 ¹³⁷Cs activity concentration levels were still present in perch and pike. Since 1989 their ¹³⁷Cs activity concentrations in oligohumic seepage lakes have remained significantly higher than in polyhumic drainage lakes due to the increased transfer of ¹³⁷Cs into fish in the seepage lakes with lower electrolyte concentrations, longer water retention times and lower sedimentation rate.     

Catalytic gasification of tars from a dumping site

The work deals with catalytic gasification, pyrolysis and non-catalytic gasification of tar from an industrial dumping site. All experiments were carried out in a vertical stainless steel gasification reactor at 800 °C. Crushed calcined dolomite was used as the gasification catalyst. Parameters such as addition of water and air, and the influence of the catalyst in regard to the composition of the process gas were investigated. The catalytic gasification experiment in the steady state produced process gas with the composition: 56 % of H₂, 9 % of CO, 11 % of CH₄ and 12 % of CO₂ (mol.%). Concentration of the C₂ fraction was lower than 1 mol.%. Volume flow of air was later changed from 120 to 230 ml min^?1 reducing the amount of hydrogen to 51 mol.% and that of methane to 10 mol.%. Process gas created in a non-catalytic gasification process contained 26–30 mol.% of methane, 13–15 mol.% of carbon monoxide and 15–17 mol.% of the C₂ fraction and lower amounts of hydrogen (20 mol.%) and carbon dioxide (2–3 mol.%). The highest apparent conversion of tar was reached in the catalytic gasification processes. A higher rate of catalyst deactivation can be observed when water or air is not added.     

Formation of PFOA from 8:2 FTOH in closed-bottle experiments with brackish water

The formation of perfluorooctanoate (PFOA) from 1H,1H,2H,2H-perfluorodecanol (8:2 FTOH) was studied for the first time in laboratory experiments with brackish water. The water samples were collected from the Baltic Sea, which is one of the largest brackish water areas in the world and is polluted with PFOA and other perfluorinated compounds. The formation of PFOA was studied in closed-bottle experiments at different water temperatures. As a reference experiment, a modified OECD 310 test was conducted with sludge from a wastewater treatment plant and with brackish water. The PFOA and 8:2 FTOH were concentrated from water samples by solid-phase extraction (SPE) and were analysed using liquid chromatography–mass spectrometry. The effect of oxygen concentration on the formation of PFOA was studied using surface water samples with high and low oxygen contents. Other experiments were performed with oxygen-rich surface water and oxygen-deficient bottom water. The formation of PFOA was observed in all experiments; it was higher in the trial performed with brackish water than in the reference test carried out with sludge. Clear temperature dependence was observed in the formation of PFOA in brackish water tests; after a 30-day test period, a sixfold increase was observed in the amount of PFOA in surface water between the temperatures of 15 and 20 °C. Microbes were suggested as the major cause of the formation of PFOA, but other environmental characteristics, such as oxygen, could also affect the formation potential of PFOA.     

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.     

Heavy-duty,off-road diesel engine low-load particle number emissions and particle control

Exhaust gas particle and ion size distributions were measured from an off-road diesel engine complying with the European Stage IIIB emission standard. The measurements were performed at idling and low load conditions on an engine dynamometer. Nucleation-mode particles dominated the diesel exhaust particle number emissions at idle load. The nonvolatile nucleation-mode geometric mean diameter was detected at 10 nm or below. The nonvolatile nucleation-mode charge state implied that it has evolved through a highly ionizing environment before emission from the engine. The determined charging probabilities were 10.0 ± 2.2% for negative and 8.0 ± 2.0% for positive polarity particles. The nonvolatile nucleation particle concentration and size was also shown to be dependent on the lubricant oil composition. The particle emissions were efficiently controlled with a partial filter or with partial filter + selective catalytic reduction (SCR) combination. The particle number removal efficiencies of the aftertreatment systems were over 95% for wet total particle number (>3nm) and over 85% for dry particle total number. Nevertheless, the aftertreatment systems’ efficiencies were around 50% for the soot-mode particles. The low-load nonvolatile nucleation particle emissions were also dependent on the engine load, speed, and fuel injection pressure. The low load particle number emissions followed the soot-core trade-off, similar to medium or high operating loads.

Implications:Idling and low-load diesel engine exhaust emissions affect harmfully the ambient air quality. The low-load particle number emissions are here shown to peak in the 10-nm size range for a modern off-road engine. The particles are electrically charged and nonvolatile and they originate from the combustion process. Tailpipe particle control by open channel filter can remove more than 85% of the nonvolatile 10-nm particles and about 50% of the soot-mode particles, while the fuel injection pressure increase can lead to particle number increase. The study provides a new viewpoint for low-load particle emissions and control.     

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.