Proposal for selective differentiating of nerve agent G and V type with utilisation of modified Ellman’s method

We studied issues of organophosphorus agents’ analysis. Immobilised enzyme–inhibitors complexes (e.g. acetylcholinesterase-organophosphate nerve agent) were studied with modified Ellman’s biochemical method utilised for assessment of acetylcholinesterase activity. Biochemical reactions are widespread and they are the most frequent used analytical methods for determination of nerve agents. This modified method is based on the nucleophilic reactions of mono- and bispyridinium aldoximes of a type 2-PAM, MMB-4 and HI-6, their homologues and isomers with enzyme–inhibitor complexes. The procedure for a gradual analysis of G and V type, Sarin, Cyclohexylsarin, Soman, Tabun, agent VX and R-33 was proposed in terms of studied nucleophilic substitution reactions quantification results. This method enables selective determination of these chemical warfare agents. A gradual analysis was evaluated by statistic method of probabilistic calculus. This type of analysis can be used for assessment of acetylcholinesterase inhibitors in very low concentrations close to hygienic limits.     

Distribution of Redox-sensitive Elements in Bottom Waters,Porewaters and Sediments of Rogoznica Lake (Croatia) in Both Oxic and Anoxic Conditions

Geochemical, mineralogical and sedimentological analyses were carried out to contrast two different sites (respectively characterized by permanently oxic and anoxic conditions) in a small, meromictic, seawater lake. In fact, due to relatively high organic matter content, and reduced water exchange, the Rogoznica Lake has almost permanent anoxic conditions below the depth of 12 m, where sediment can be considered an anoxic–sulphidic sedimentary environment. Different water column and sediments redox conditions affect the distribution and speciation of major redox-sensitive metals (Fe, Mn, Mo), reduced sulphur species (RSS) and dissolved organic C (DOC). Trace metals, especially those that accumulate in anoxic–sulphidic environments (Fe, Mo) showed a marked enrichment in the solid phase, whereas the low solubility of sulphides leads to low porewater concentrations. The relatively high sedimentary enrichment of Mo (up to 81 mg/kg) also confirms highly anoxic conditions within the Rogoznica Lake sediments. Results clearly show that chemical species within the sediments will tend towards equilibrium between porewater and solid phase according the prevailing environment conditions such as redox, pH, salinity, DOC.     

Procedures of trophic chain samples preparation for determination of triazines by HPLC and metals by ICP-AES methods

The aim of this research was monitoring the distribution of atrazine and simazine as well as metals Pb, Cd, Zn, Al, Co, Ni, and V along with trophic chains: soil-vegetables and soil, carrot or grass and meat. Different techniques of herbicides extraction by means of many solvents were examined. Triazines were analysed by means of HPLC, metals by means of ICP-AES. Detection limits: LOD=0.2 microg ml(-1), determination limits: LOQ=0.73 microg ml(-1) for atrazine and LOD=0.3 microg ml(-1), LOQ=1.12 microg ml(-1) for simazine were obtained. The content (microg g(-1)) of simazine in soil was in range: 3.45-8.60, in vegetable roots: 6.62-38.15, in vegetable leaves: 2.45-31.71, in rabbit fat: 0.13-49.90. The content (microg g(-1)) of atrazine in soils was in range: 11.9-13.03, in vegetable roots: 13.61-92.90. In analysed material the particular metals after microwave or dry digestion were determined in range (microg g(-1)): Pb: 6.48-43.18; Cd: 0.11-0.57; Zn: 8.79-51.90; Al: 10.22-24.48; Co: 0.18-3.89; Ni: 0.37-6.36; V: 0.29-1.48.     

Chemical composition and hygroscopic properties of size-segregated aerosol particles collected at the Adriatic coast of Slovenia

The chemical composition as well as the water uptake characteristics of aerosols was determined in size-segregated samples collected during November 2002 on the Slovenian coast. Major ions, water-soluble organic compounds (WSOC), short-chain carboxylic acids and trace elements were determined in the water-soluble fraction of the aerosol. Total aerosol black carbon (BC) was measured from filter samples. Our results showed that the origin of air masses is an important factor that controls the variation in the size distribution of the main components. Very high concentrations of WSOC as well as higher concentrations of BC were found under mostly continental influence. Besides the main ionic species (SO4(2-), NH4(+), K+) in the finest size fraction (0.17-0.53 microm), the concentration of NO3(-) was also high. The difference between the two different air mass origins is particularly expressed for Cl-, Na+, Mg2+ and Ca2+ determined in particles larger than 1.6 microm. As expected, a very good correlation was found between Na+ and Cl-. A good correlation was found between sea salt elements and elements of crustal origin (Na+, Cl-, Mg2+, Ca2+, Sr). A good relationship between typical anthropogenic tracers (K, V and Pb) was also observed. The mass growth factors, for all size fractions of aerosols collected under continental influence were very low (maximum 2.23 at 94%, 1.6-5.1 microm), while under marine influence the mass growth factors increased significantly with the particle size. At 97% humidity, the mass growth factors were 6.95 for the size fraction 0.53-1.6 microm and 9.78 for larger particles (1.6-5.1 microm).     

Chromosomal aberrations in ovine lymphocytes exposed in vitro to tolylfluanid

Chromosomal aberrations have been used as important cytogenetic biomarkers to study the mutagenic effects of different chemicals in vivo and in vitro. Chromosomal aberrations were evaluated in cultures of sheep lymphocytes in vitro exposed to the fungicide tolylfluanid. Lymphocyte cultures from three donors were exposed to four different concentrations of fungicide (1.10(-4) M(.)L; 1.10(-5) M(.)L; 1.10(-6) M(.)L; 1 × 10(-7) M(.)L). Chromosomal analysis showed a significant (P = 0.018 and 0.038 respectively, Anova test, P < 0.05, Tukey test) increase in the frequency of aberrant cells (ABC) in cultures treated with the highest negative experimental concentrations of tolylfluanid (1.10(-4) M(.)L; 1.10(-5) M(.)L) compared to control. Significantly increased numbers of chromatid breaks (7.67 ± 0.58% against 1.67 ± 2.08%, P = 0.009, Anova test, P < 0.05, Tukey test) and chromatid gaps (7.67 ± 1.15% against 2.67 ± 0.58%, P?= 0.003, Anova test, P < 0.05, Tukey test) were observed in ovine cultures treated with the highest experimental concentration of tolylfluanid (1.10(-4) M(.)L). Tolylfluanid induced also chromosomal exchanges (P = 0.038, and 0.016 respectively, Anova test, P < 0.05, Tukey test) in ovine cultures treated with the highest experimental concentrations of tolylfluanid (1.10(-4) M(.)L; 1.10(-5) M(.)L). The mitotic index has not shown any statistical differences between the various treatments and control groups. Our results suggest a significant genotoxic effect of tolylfluanid only at the highest concentration in sheep peripheral lymphocytes in vitro.     

Carbon exchange between ecosystems and atmosphere in the Czech Republic is affected by climate factors

By comparing five ecosystem types in the Czech Republic over several years, we recorded the highest carbon sequestration potential in an evergreen Norway spruce forest (100%) and an agroecosystem (65%), followed by European beech forest (25%) and a wetland ecosystem (20%). Because of a massive ecosystem respiration, the final carbon gain of the grassland was negative. Climate was shown to be an important factor of carbon uptake by ecosystems: by varying the growing season length (a 22-d longer season in 2005 than in 2007 increased carbon sink by 13%) or by the effect of short- term synoptic situations (e.g. summer hot and dry days reduced net carbon storage by 58% relative to hot and wet days). Carbon uptake is strongly affected by the ontogeny and a production strategy which is demonstrated by the comparison of seasonal course of carbon uptake between coniferous (Norway spruce) and deciduous (European beech) stands.     

Nitrogen removal from wastewater and bacterial diversity in activated sludge at different COD/N ratios and dissolved oxygen concentrations

The impact of the organic carbon to nitrogen ratio (chemical oxygen demand (COD)/N) in wastewater and dissolved oxygen (DO) concentration on carbon and nitrogen removal efficiency, and total bacteria and ammonia-oxidizing bacteria (AOB) communities in activated sludge in constantly aerated sequencing batch reactors (SBRs) was determined. At DO of 0.5 and 1.5 mg O₂/L during the aeration phase, the efficiency of ammonia oxidation exceeded 90%, with nitrates as the main product. Nitrification and denitrification achieved under the same operating conditions suggested the simultaneous course of these processes. The most effective nitrogen elimination (above 50%) was obtained at the COD/N ratio of 6.8 and DO of 0.5 mg O₂/L. Total bacterial diversity was similar in all experimental series, however, for both COD/N ratios of 6.8 and 0.7, higher values were observed at DO of 0.5 mg O₂/L. The diversity and abundance of AOB were higher in the reactors with the COD/N ratio of 0.7 in comparison with the reactors with the COD/N of 6.8. For both COD/N ratios applied, the AOB population was not affected by oxygen concentration. Amplicons with sequences indicating membership of the genus Nitrosospira were the determinants of variable technological conditions.     

Effect of biosurfactants on crude oil desorption and mobilization in a soil system

Microbially produced biosurfactants were studied to enhance crude oil desorption and mobilization in model soil column systems. The ability of biosurfactants from Rhodococcus ruber to remove the oil from the soil core was 1.4-2.3 times greater than that of a synthetic surfactant of suitable properties, Tween 60. Biosurfactant-enhanced oil mobilization was temperature-related, and it was slower at 15 degrees C than at 22-28 degrees C. Mathematical modelling using a one-dimensional filtration model was applied to simulate the process of oil penetration through a soil column in the presence of (bio)surfactants. A strong positive correlation (R(2)=0.99) was found between surfactant penetration through oil-contaminated soil and oil removal activity. Biosurfactant was less adsorbed to soil components than synthetic surfactant, thus rapidly penetrating through the soil column and effectively removing 65-82% of crude oil. Chemical analysis showed that crude oil removed by biosurfactant contained a lower proportion of high-molecular-weight paraffins and asphaltenes, the most nonbiodegradable compounds, compared to initial oil composition. This result suggests that oil mobilized by biosurfactants could be easily biodegraded by soil bacteria. Rhodococcus biosurfactants can be used for in situ remediation of oil-contaminated soils.     

Iron-catalyzed oxidation of s(IV) species by oxygen in aqueous solution: Influence of pH on the redox cycling of iron

Redox cycling of Fe(II)/Fe(III) during the catalyzed aqueous S(IV) oxidation by dissolved oxygen in the presence of Fe(II) and/or Fe(III) at an initial pH_i of 4.4, often observed in atmospheric waters, was studied in detail. It has been found that the reaction rate is not considerably affected by the oxidation state of iron at the start of the reaction. An equilibrium between Fe(II) and Fe(III) was established a few minutes after the start of the reaction, regardless of the oxidation state of iron at the beginning of the experiment. The prevailing oxidation state of iron in a particular phase of the reaction depends on the concentration of S(IV) in the reaction solution. It has been found that the formation of polymerized hydroxo Fe(III) species is also included in the mechanism of the Fe-catalyzed oxidation of S(IV). The formation of these species was confirmed by the on-line measurement of Fe(II) and Fe_reac. The results also clearly demonstrate that the pH_i of the solution is a major factor, controlling the concentration of Fe(III) ions, the form of S(IV) species, and consequently the reaction rate of S(IV) oxidation by oxygen.