A comparison of biochemical responses to oxidative and metal stress in seedlings of barley, Hordeum vulgare L

Biochemical responses on the bases of activities of antioxidant enzymes; peroxidase, catalase, superoxide dismutase and glutathione reductase as well as estimations of total protein, lipid peroxidation and thiols in the form of protein, non-protein, glutathione and phytochelatin measured in growing seedlings of barley, Hordeum vulgare L., from Day 2 through 8 were compared following treatment of seeds for 2 h with oxidative agents, paraquat 5 x 10(-5), 10(-4), 10(-3) M, H2O2 10(-3), 5 x 10(-3), 10(-2) M and a metal salt, CdSO4 10(-5), 10(-4), 10(-3) M. A significant induction of all antioxidant enzymes along with an increase in the levels of protein, lipid peroxidation and glutathione was noted in response to oxidative stress, CdSO4 induced significant peroxidase and catalase activities but not superoxide dismutase. In a marked contrast from oxidative stress, CdSO4 decreased glutathione reductase activity as well as glutathione levels but increased phytochelatin level. The differential biochemical responses thus underlined the crucial involvement of glutathione and phytochelatin in the oxidative and metal-induced adaptive responses, respectively.     

Effects of arsenic on blast transformation and DNA synthesis of human blood lymphocytes

Effects of inorganic arsenicals on DNA synthesis in unsensitized human blood lymphocytes were biphasic: the chemicals at very low concentrations enhanced blast transformation and DNA synthesis, whereas higher concentrations inhibited the transformation and DNA synthesis. The concentrations of arsenicals at which the maximum stimulating effect was found were 1 x 10(-5) M, 1 x 10(-6) M or 2 x 10(-6) M, and 0.8 x 10(-6) M or 1 x 10(-6) M for sodium arsenite exposure of 1 h, 3 days and 6 days, respectively; for sodium arsenate, 1 x 10(-5) M, 1 x 10(-5) M, and 2 x 10(-6) M or 5 x 10(-6) M, respectively. Arsenicals must be present for the entire 6 days culture period to produce maximum stimulation of blast transformation of human lymphocytes. The longer exposure of the lymphocytes to arsenicals, the lower the concentrations of arsenicals at which the maximum stimulating effect was found. The stimulating effect of trivalent arsenic (sodium arsenite) was stronger than pentavalent arsenic (sodium arsenate).     

Chronic toxicity of copper on embryo development in Chinese toad, Bufo gargarizans

This study examined the effects of copper exposure on embryonic development of Chinese toad, Bufo gargarizans. Firstly, the LC₅₀ values from 24 to 96 h of exposure were 3.61 × 10⁻⁶ M, by means of a 4 d toxicity test with B. gargarizans embryos. Secondly, Chinese toad embryos were exposed to 10⁻⁹-10⁻⁶ M copper from mid gastrula stage to operculum completion stage. Measurements included mortality, tadpole weight, tadpole total length, growth retardation, duration of different embryo stages and malformation. Embryonic survival was not affected by copper. Relative to control tadpoles, significantly decreased weight and total length were found at 10^-9-10^-6 M reduced percentage of the embryos in right operculum stage after 10 d exposure to copper and reduced percentage of embryos in operculum completion stage after 12 d exposure to copper were also observed. Moreover, the duration of embryonic development increased at neural, circulation and operculum development stage in copper-treated groups. For the scanning microscope and histological observation, the abnormalities were malformation of wavy dorsal fin, flexural tail, curvature body axis, yolk sac oedema and reduced pigmentation in the yolk sac. Histopathological changes in olfactory, retinal epithelium and skin were also observed. DNA strand breaks exposed to the copper were analyzed by DNA ladder. In conclusion, copper induced toxic effects on B. gargarizans embryos. The present study indicated chronic toxicity tests may provide more accurate way in formulating the “safe levels” of heavy metals to amphibian.     

Effects of cold-dark storage on growth of Cylindrotheca closterium and its sensitivity to copper

Cylindrotheca closterium cells were maintained at low temperature (4+/-1 degrees C) and dark conditions up to 21 weeks to assess the effect on survival and physiological status. From a control culture under standard conditions, three densities were prepared: (A) 2 x 10(4), (B) 10 x 10(4), and (C) 25 x 10(4) cells ml(-1). Weekly, inoculums of each stored density were exposed to continuous light and at 20+/-1 degrees C. Sensitivity to copper for microalgal cultures was evaluated in order to assess possible changes in cells sensitivity due to storage. Concurrently, assays with a control culture were carried out in order to assess the sensitivity of C. closterium to copper and to be able to generate a standard sensitivity control chart with a mean value of EC50-72 h+/-2SD (standard deviation). Density-C presented higher cell yield values, between 40% and 80% relative to control culture. Cell density showed to be important feature that may be taken into account in cell storage experiments. There was an increase in sensitivity of cells submitted to storage; however results always kept in the range established as standard sensitivity with no statistically significant difference with regards to control culture. EC50-72 h mean value for the control culture was 29+/-10 mug Cul(-1), while for densities-A, B and C were 22+/-7; 23+/-9 and 23+/-8 microg Cul(-1), respectively. In spite of drastic changes in the environmental conditions due to storage, it is concluded that C. closterium cells stored during 5 months remained metabolically active and with no significant change in its sensitivity.     

Homogeneous degradation of 1,2,9,10-tetrachlorodecane in aqueous solutions using hydrogen peroxide, iron and UV light

The homogeneous degradation of the polychlorinated n-alkane, 1,2,9,10-tetrachlorodecane (T4C10), was studied in aqueous solutions of hydrogen peroxide, including Fenton and photo-Fenton reaction conditions. All solutions were adjusted to a pH of 2.8 and an ionic strength of 0.1 M NaClO4 prior to photolysis. T4C10 (2 x 10(-6) M) was substantially degraded by the H2O2/UV system (1.0 x 10(-2) M H2O2), with 60% disappearance in 20 min of irradiation in a photoreactor equipped with 300 nm lamps of light intensity 3.6 x 10(-5) Ein L(-1) min(-1) (established by ferrioxalate actinometry). The reaction produced stoichiometric amounts of chloride ion indicating complete dechlorination of the chlorinated n-alkane. T4C10 degraded very slowly under Fenton (Fe2+/H2O2/dark) and Fenton-like (Fe3+/H2O2/dark) conditions. However, when the same solutions were irradiated, T4C10 degraded more rapidly than in the H2O2/UV system, with 61% disappearance in 10 min of exposure. The rapid degradation is related to the enhanced degradation of hydrogen peroxide to oxidizing *OH radicals under photo-Fenton conditions. Degradation was inhibited in both the H2O2/UV and photo-Fenton systems by the addition of KI and tert-butyl alcohol due to *OH scavenging.     

Fractionation of fulvic acids: characteristics and complexation with copper

Two fulvic acid samples isolated from Suwannee river (International Humic Substance Society) and Feeitsuey reservoir were subjected to gel filtration chromatography (GFC) for molecular size fractionation. The GFC-eluted samples were separated into three groups corresponding to the molecular weight ranges: < 220, 220-1000, and 1000-4000. Fluorescence quenching techniques were employed for determining the conditional stability coefficient and kinetic parameters of copper complexation with the three fractions of fulvic acids. Experimental conditions were pH 6, 5 x 10(-5)m total copper and 5 mg C litre(-1) of fulvic acids. The conditional stability coefficients of the fulvic acid fractions were in the order of 0.9-3.3 x 10(5)m(-1), and the forward and reverse rate constants were in the order of 6.9-12.4 x 10(3)m(-1) s(-1) and 3.5-8.0 x 10(-2) s(-1). Information could be useful in modelling copper transport in the hydrosphere.     

Effects of ionic strength on the production of short chain volatile hydrocarbons by Dunaliella salina (Teodoresco)

The effect of ionic strength on the production of short chain volatile hydrocarbons was studied in cultures of Dunaliella salina. Axenic cultures of D. salina were grown at three different ionic strengths 0.5, 2 and 3 M of NaCl in Johnson (J/1) culture medium [Journal of Bacteriology 95 (1968) 1461] under the following laboratory growth conditions: a 12:12 h photoperiod, 300 micromolm(-2)s(-1) of photosynthetic active radiation (PAR) provided by a fluorescent lamp of 40 W combined with a 100 W incandescent lamp at 20 +/- 1 degrees C at pH 7.5. C1 to C5 hydrocarbons were detected using a head space technique and GC-FID. Cell numbers and growth rate was greatest at 2 M NaCl 4.3 x 10(6) cellml(-1) after a 15 days period of culture. Maximum hydrocarbon production was measured in the concentration of 0.5 NaCl with lower production rates in the more concentrated solutions. The principal hydrocarbon was pentane at 0.5 M but was ethane in 2 and 3 M solutions. Production rates for individual compounds ranged between 0.13 and 22 x 10(-15) microgCcell(-1)h(-1). It is suggested that the ability to produce and release volatile organic compounds of D. salina is related to osmotic conditions established by the ionic strength of growth solution.     

Kinetics and mechanism of nitrite oxidation by hypochlorous acid in the aqueous phase

The rate coefficient for the reaction of nitrite with hypochlorite and hypochlorous acid has been studied using spectrophotometric measurements. The reaction rate has been determined in a wide range of H(+) concentration (5< or =-log[H(+)]< or =11). The kinetics were carried out as a function of NO(2)(-), H(+) and total hypochlorite ([HOCl](total)=[HOCl]+[ClO(-)]+[ClNO(2)]) concentrations. The observed overall rate law is described by: -d[HClO](T)dt=[a[NO(2)(-)](2)+b[NO(2)(-)]][H(+)](2)c+d[H(+)]+e[NO(2)(-)][H(+)](2)[HOCl](total)At T=298 K and in Na(2)SO(4) at an ionic strength (I=1.00 M), we obtained using a nonlinear fitting procedure: a=(1.83+/-0.36)x10(7) s(-1), b=(1.14+/-0.23)x10(5) Ms(-1), c=(1.12+/-0.17)x10(-13) M, d=(1.43+/-0.29)x10(-6) M(2) and e=(1.41+/-0.28)x10(3) M where the errors represent 2sigma. According to the overall rate law, a/b=k(1)/k(3), b/e=k(3), c=K(w), d/c=K(a), d=K(a)K(w) and e=K(1)K(a). In Na(2)SO(4) at an ionic strength (I=1.00 M), the values of K(1) and K(a) are (1.1+/-0.1)x10(-4) and 1.28x10(7) M(-1), respectively. A mechanism is proposed for the NO(2)(-) oxidation which involves the reversible initial step: NO(2)(-)+HOCl left harpoon over right harpoon ClNO(2)+OH(-) (K(1)), while ClNO(2) undergoes the two parallel reactions: attack by NO(2)(-) (k(1)) and hydrolysis (k(3)). ClNO(2) and N(2)O(4) are proposed as important intermediates as they control the mechanism. The rate coefficients k(1) and k(3) have been determined at different ionic strengths in NaCl and Na(2)SO(4). The influence of the ionic strength and ionic environment has been studied in this work.     

The characteristics of wet air oxidation of phenol over CuOx/Al2O3 catalysts: effect of copper loading

The nature of active copper species is well-known to vary with copper loading, i.e., isolated Cu(2+) to bulk CuO. In this work, however, the effect of copper loading on the activity and the selectivity was investigated for the wet oxidation of phenol over CuO(x)/Al(2)O(3) catalysts. The activity and the mineralization selectivity of the catalysts increased with copper loading up to 7wt% and remained almost the same at a higher loading. The optimum copper loading was about 7wt% for the wet oxidation of phenol over CuO(x)/Al(2)O(3) catalysts in this work. The nature of copper species with different loading was characterized with TPR, XRD, and XANES. The chemical states of copper in the CuO(x)/Al(2)O(3) catalysts were confirmed as varying with copper loading: isolated Cu(2+) ions for 1wt%; highly dispersed Cu(2+) cluster for 5wt% and 7wt%, and bulk CuO for 10-25wt%. The stability of the CuO(x)/Al(2)O(3) catalysts with different copper loading was also studied with respect to carbonaceous deposits and copper leaching.     

Effect of plant growth on copper solubility and speciation in soil solution samples

The effect of plant growth on copper solubility and speciation was studied in a 10-week pot experiment. A copper-tolerant grass variety (Agrostis capillaris L. var. Parys Mountain) was grown in pots that contained either clean (copper-total approx. 30 mg kg(-1)) or copper contaminated soil (copper-total approx. 170 mg kg(-1)) at two pH levels (4.7 and 5.5). Also, similar pots without vegetation were included in the study. Due to the addition of NH(4)NO(3) fertilizer and subsequent nitrification of ammonia to nitrate, soil pH decreased from 4.7 to 3.5 and from 5.5 to 4, respectively. In the planted pots, soil pH recovered faster after depletion of NH(4)(+). This resulted in a decrease in the calcium solution concentrations and an increase in the dissolved organic carbon (DOC) concentrations in the planted pots. However, this was only observed in the clean soil; in the contaminated soil no difference in DOC levels between bare and planted pots was observed. Copper solubility in the contaminated soil was lower in the presence of plants; in the clean soil no differences were observed between the bare and planted pots. In the planted pots, copper activities in solution in both clean and contaminated soils were two orders of magnitude lower than in the bare pots. Copper activities in the non-planted contaminated soil reached potentially toxic levels ([Cu]+/-10(-5) to 10(-6) M) in contrast to the lower levels in the planted pots ([Cu]+/-10(-7) to 10(-10) M). Data and model results show that plant growth improves pH, DOC and calcium in solution to such an extent that both the total dissolved copper concentration and the free metal activity in soils can be reduced. This stresses the potential beneficial role of plants for the immobilization and detoxification of metals in contaminated soils.     

Uptake of 134Cs in the shoots of Amaranthus tricolor and Amaranthus cruentus

Amaranthus tricolor L. and Amaranthus cruentus L. were grown in pots containing 7.5 kg soils artificially contaminated with three levels of 134Cs activity: 5.55 x 10(5) Bq pot-1, 1.11 x 10(6) Bq pot(-)1, and 1.665 x 10(6) Bq pot(-1), respectively. Forty-nine days after sowing and growth, plants were harvested. The plants growing in soils with increasing 134Cs concentrations showed increasing concentration of this radionuclide in shoots. There were significant differences in uptake of 134Cs applied to soils between and within the plant species, depending on the initial 134Cs concentrations. The plant species showed different responses to the addition of (NH4)2SO4 to soils. Biomass production of both species was reduced in pots treated with (NH4)2SO4. (NH4)2SO4 application decreased the uptake of 134Cs by A. tricolor but increased the accumulation of 134Cs by A. cruentus, showing that chemicals with the highest efficiency to enhance the desorption of 134Cs might play an unexpected role in transferring the radionuclide to shoots.     

Selenium oxyanion inhibition of hydrogenotrophic and acetoclastic methanogenesis

Inhibitory effects of selenite and selenate towards hydrogenotrophic and acetoclastic methanogenesis were evaluated in anaerobic toxicity assays. The 50% inhibitory concentration (IC(50)) of both selenium oxyanions was below 6.1x10(-5)M in hydrogenotrophic assays, whereas acetoclastic methanogens were less inhibited: IC(50)=8.3x10(-5)M and 5.5x10(-4)M for selenite and selenate, respectively. Selenite completely inhibits methanogenesis from both substrates tested at concentrations 10(-3)M selenite, while only marginal methanogenic activities occur at equimolar concentrations of selenate. Selenite becomes irreversibly inhibitory upon a single exposure, whereas selenate inhibits methanogens upon repeated exposure. Consequently, methane recovery can be seriously hampered or even impossible during anaerobic treatment of highly selenium contaminated waste streams.     

Oxidation of MCPA and 2,4-D by UV radiation, ozone, and the combinations UV/H2O2 and O3/H2O2

The phenoxyalkyl acid derivative herbicides MCPA (4-chloro 2-methylphenoxyacetic acid) and 2,4-D (2,4-dichlorophenoxyacetic acid) were oxidized in ultrapure water by means of a monochromatic UV irradiation and by ozone, as well as by the combinations UV/H2O2 and O3/H2O2. In the direct photolysis of MCPA, the quantum yield at 20 degrees C was directly evaluated and a value of 0.150 mol Eins(-1) was obtained in the pH range 5-9, while a lower value of 0.41 x 10(-2) mol Eins(-1) was determined at pH=3. Similarly, for 2,4-D a value of 0.81 x 10(-2) mol Eins(-1) was deduced, independent of the pH of work. The influence of the additional presence of hydrogen peroxide was established in the combined process UV/H2O2, and the specific contribution of the radical pathway to the global photo-degradation was evaluated. The oxidation by ozone and by the combination O3/H2O2 was also studied, with the determination of the rate constants for the reactions of both herbicides with ozone and hydroxyl radicals at 20 degrees C. These rate constants for the direct reactions with ozone were 47.7 and 21.9 M(-1) s(-1) for MCPA and 2,4-D respectively, while the found values for the rate constants corresponding to the radical reactions were 6.6 x 10(9) and 5.1 x 10(9) M(-1) s(-1).     

Micronucleus distribution in human peripheral blood lymphocytes treated in vitro with cadmium chloride in G0 and S phase of the cell cycle

Cadmium chloride (CdCl2 x H2O) in concentrations 10(-3) - 10(-6) M was tested for genotoxicity in human lymphocytes in vitro. The DNA damage was expressed through the occurrence of micronuclei (MN) and was detected using the cytochalasin-B-blocked MN assay. Human blood was treated in the G0 and S phase of the cell cycle. All except the highest concentration of cadmium chloride of 10(-3) M applied in the G0 phase of the cell cycle resulted in the increase in MN cells, but it was not statistically significant. Cadmium chloride added to the cultures in the concentration of 10(-3) M affected the cell growth regardless of the phase. Cadmium chloride added to cultures 24 h after their initiation (early S phase) was found to significantly increase the MN frequency in 10(-4) - 10(-6) M concentrations (P > 0.05).     

Kinetic assessment of the potassium ferrate(VI) oxidation of antibacterial drug sulfamethoxazole

Sulfamethoxazole (SMX), a worldwide-applied antibacterial drug, was recently found in surface waters and in secondary wastewater effluents, which may result in ecotoxical effects in the environment. Herein, removal of SMX by environmentally-friendly oxidant, potassium ferrate(VI) (K(2)FeO(4)), is sought by studying the kinetics of the reaction between Fe(VI) and SMX as a function of pH (6.93-9.50) and temperature (15-45 degrees C). The rate law for the oxidation of SMX by Fe(VI) is first-order with respect to each reactant. The observed second-order rate constant decreased non-linearly from 1.33+/-0.08 x 10(3) M(-1)s(-1) to 1.33+/-0.10 x 10(0) M(-1)s(-1) with an increase of pH from 7.00 to 9.50. This is related to protonation of Fe(VI) (HFeO(4)(-) <==> H(+) + FeO(4)(2-); pK(a,HFeO(4)) = 7.23) and sulfamethoxazole (SH <==> H(+) + S(-); pK(a,SH)=5.7). The estimated rate constants were k(11)(HFeO(4)(-) + SH) = 3.0 x 10(4) M(-1)s(-1), k(12)(HFeO(4)(-) + S(-)) = 1.7 x 10(2) M(-1)s(-1), and k(13) (FeO(4)(2-) + SH) = 1.2 x 10(0) M(-1)s(-1). The energy of activation at pH 7.0 was found to be 1.86+/-0.04 kJ mol(-1). If excess potassium ferrate(VI) concentration (10 microM) is used than the SMX in water, the half-life of the reaction using a rate constant obtained in our study would be approximately 2 min at pH 7. The reaction rates are pH dependent; thus, so are the half-lives of the reactions. The results suggest that K(2)FeO(4) has the potential to serve as an oxidative treatment chemical for removing SMX in water.     

Pd-nanoparticles cause increased toxicity to kiwifruit pollen compared to soluble Pd(II)

In the present study, endpoints including in vitro pollen performance (i.e., germination and tube growth) and lethality were used as assessments of nanotoxicity. Pollen was treated with 5-10 nm-sized Pd particles, similar to those released into the environment by catalytic car exhaust converters. Results showed Pd-nanoparticles altered kiwifruit pollen morphology and entered the grains more rapidly and to a greater extent than soluble Pd(II). At particulate Pd concentrations well below those of soluble Pd(II), pollen grains experienced rapid losses in endogenous calcium and pollen plasma membrane damage was induced. This resulted in severe inhibition and subsequent cessation of pollen tube emergence and elongation at particulate Pd concentrations as low as 0.4 mg L⁻¹. Particulate Pd emissions related to automobile traffic have been increasing and are accumulating in the environment. This could seriously jeopardize in vivo pollen function, with impacts at an ecosystem level.     

Kinetics of hydrolysis and cyclization of ethyl 2-(aminosulfonyl)benzoate to saccharin

The cyclization of ethyl 2-(aminosulfonyl)benzoate (ASB) to give saccharin was investigated in aqueous solutions at pH between 5.2 and 9.5 and in the temperature range of 296.2-334.2 K. The initial concentration of the reactant was varied between 1.45 x 10(-5) and 3.86 x 10(-4) M. Ultraviolet spectroscopy was used to obtain the kinetic data. The reaction is acid catalyzed and follows pseudo-first-order kinetics. The experimental rate constant, k(obs), increases with temperature and pH. Its dependence on the temperature and pH is well described by: k(obs) = k1 [OH-] = [(2.52 +/- 0.9) x 10(16) exp(-20.2 +/- 1 kcalmol(-1)/RT) s(-1)][OH-] A mechanism is proposed and the half-life of ethyl ASB is calculated.     

Inhibitory effect of ragweed (Ambrosia artemisiifolia L)-inflorescence extract on the germination of Amaranthus hypochondriacus L and growth of two soil algae

In this study the ecological importance of common ragweed (Ambrosia artemisiifolia L.; Asteraceae) was investigated in in vitro bioassays as it is one of the most widespread annual weeds occurring on arable land in Hungary. Eight fractions were derived from ragweed inflorescence water extract by low vacuum column chromatography and their biological activity was investigated by Amaranthus hypochondriacus L. cv. Edit (Amaranthaceae) germination bioassay. As a result of the germination bioassay the chromatographic method was modified to harvest the most effective fractions in only one fraction, which was used in three concentration (10, 50, 100 mg x kg(-1)) for algal growth inhibition bioassay.Results showed significant growth inhibition in case of both applied algal species.     

Effects of copper deficiency and copper toxicity on organogenesis and some physiological and biochemical responses of Scots pine (Pinus sylvestris L.) seedlings grown in hydroculture

The morphological, physiological, and biochemical parameters of 6-week-old seedlings of Scots pine (Pinus sylvestris L.) were studied under deficiency (1.2 nM) and chronic exposure to copper (0.32, 1, 2.5, 5, and 10 μM CuSO₄) in hydroculture. The deposit of copper in the seed allowed the seedlings to develop under copper deficiency without visible disruption of growth. The high sensitivity of Scots pine to the toxic effects of copper was shown, which manifested as a significant inhibition of growth and development. The loss of dominance of the main root and a strong inhibition of lateral root development pointed to a lack of adaptive reorganization of the root system architecture under copper excess. A preferential accumulation of copper in the root and a minor translocation in aerial organs confirmed that Scots pine belongs to a group of plants that exclude copper. Selective impairment in the absorption of manganese was discovered, under both deficiency and excess of copper in the nutrient solution, which was independent of the degree of development of the root system. Following 10 μM CuSO₄ exposure, the absorption of manganese and iron from the nutrient solution was completely suppressed, and the development of seedlings was secured by the stock of these micronutrients in the seed. The absence of signs of oxidative stress in the seedling organs was shown under deficiency and excess of copper, as evidenced by the steady content of malondialdehyde and 4-hydroxyalkenals. Against this background, no changes in total superoxide dismutase activity in the organs of seedlings were revealed, and the increased content of low-molecular-weight antioxidants was observed in the roots under 1 μM and in the needles under 5 μM CuSO₄ exposures.

     

Solar light induced and TiO2 assisted degradation of textile dye reactive blue 4

Aqueous solutions of reactive blue 4 textile dye are totally mineralised when irradiated with TiO2 photocatalyst. A solution containing 4 x 10(-4) M dye was completely degraded in 24 h irradiation time. The intensity of the solar light was measured using Lux meter. The results showed that the dye molecules were completely degraded to CO2, SO4(2-), NO3-, NH4+ and H2O under solar irradiation. The addition of hydrogen peroxide and potassium persulphate influenced the photodegradation efficiency. The rapidity of photodegradation of dye intermediates were observed in the presence of hydrogen peroxide than in its absence. The auxiliary chemicals such as sodium carbonate and sodium chloride substantially affected the photodegradation efficiency. High performance liquid chromatography and chemical oxygen demand were used to study the mineralisation and degradation of the dye respectively. It is concluded that solar light induced degradation of textile dye in wastewater is a viable technique for wastewater treatment.