Micronucleus frequency in sheep lymphocytes after in vitro exposure to fungicide tolylfluanid

The fungicide tolylfluanid (N - dichlorofluoromethylthio-N′, N - dimethyl - N - p - tolylsulfamide), was investigated by cytokinesis-block micronucleus assay. Tolylfluanid at the lowest concentration (1 × 10^{? 6}mol L^{? 1})did not influence significantly the frequency of micronuclei in sheep lymphocyte cultures in comparison with control (32.33 ± 3.51/1000 binucleated cells versus 30.33 ± 2.82/1000 binucleated cells in dimethylsulfoxide (DMSO) control, P = 0.44). Higher tolylfluanid concentrations (1 × 10^{? 4} and, 1 × 10^{? 5} mol L^{? 1}) resulted in a significant dose-dependent increase in the number of micronuclei in comparison with control (74.00 ± 13.00/1000 binucleated cells and 52.67 ± 10.12/1000 binucleated cells versus 30.33 ± 2. 82/1000 binucleated cells in DMSO control, P = 0.005 and 0.02, respectively, ANOVA followed by Tukey test P < 0.05). Many of the treated cells also possessed multiple micronuclei. Tolylfluanid did not affect the nuclear division index at all treatment concentrations. Our in vitro results thus demonstrate that tolylfluanid had a significant genotoxic effect at only the highest concentration tested.     

Effect of triazole pesticide formulation on bovine culture cells

To date, most data about the possible genotoxic effect of triazole pesticides are focused on laboratory animals resulting in limited information on further non-target organisms such as cattle. The objective of the present study was to investigate the effect of triazole (tebuconazole/prothioconazole) fungicide formulation on the induction of chromosomal aberrations (CAs), sister chromatid exchanges (SCEs) and DNA fragmentation in bovine cultured lymphocytes. Our results showed that the fungicide formulation did not induce significant number of CAs in bovine cells after 24 h treatment. Nevertheless, the dose-dependent reduction of mitotic division was observed, with the strongest effect at 30.0 μg mL^?1 in both donors (P < 0.01 and P < 0.001, respectively). Prolonged 48 h exposure caused the increased level of breaks in treated cultures (3.0?15.0 μg mL^?1; P < 0.05) and significant decrease in mitotic index (MI). The tested fungicide failed to produce any statistical changes in the SCE frequency neither after 24 h nor 48 h treatment. However, the significant decline of the proliferation index (PI) was observed after 24 h indicating the fungicide influence on cell cycle kinetics. Prolonged 48 h exposure caused cytotoxicity reflecting in lower PI value relative to control mainly at the highest fungicide concentrations (30.0 μg mL^?1, P < 0.001). Using painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) only low levels of aneuploidies were detected. Significant increase of polyploidy cells (P < 0.05) was induced by a 3.0 μg mL^?1 dose of the fungicide after 48 h. DNA fragmentation assay didn't reveal the presence of DNA nucleosome ladder in cell cultures at any time (24 h and 48 h) and fungicide concentration.     

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).     

Evaluation of bendiocarb cytotoxicity in mammalian and insect cell cultures

There is an increasing need for rapid and easily interpreted in vitro assays to screen for possible cytotoxicity of pesticides. The objective of this study was to investigate the effect of the carbamate insecticide bendiocarb on mammalian and insect cell cultures. The cytotoxicity of this insecticide was evaluated by cell proliferation and cellular damage was assessed by evaluation of the cytopathic effect and lactate dehydrogenase (LDH) leakage. Cells of insect origin (Sf21) were the most sensitive to bendiocarb with significant (P < 0.01) suppression of their proliferative activity ranging from 10(-1)-10(-5) M. However, significant suppression of proliferative activity was also recorded in rat liver cells (WBF344; 10(-1)-10(-3) M; P < 0.01-0.05) and rabbit kidney cells (RK13; 10(-1) M; P < 0.01). In contrast with the proliferation activity of cells, a cytopathic effect based on cellular damage and LDH leakage into the medium was observed only at the highest concentration (10(-1) M) in RK 13 and WBF344 cells, but not in the Sf21 insect cell line. Our results indicate that bendiocarb exposure caused a cell-type dependent decrease in cell proliferation; however, cell damage and LDH leakage into the medium were not present or were strongly limited, dependent on the cell phenotype. Cell proliferation was shown as a sensitive indicator for evaluation of the cytotoxic effect of bendiocarb in vitro; on the other hand, microscopic signs of cellular damage and LDH leakage were insufficient in vitro markers.     

Evaluation of potential genotoxic/cytotoxic effects induced by epoxiconazole and fenpropimorph-based fungicide in bovine lymphocytes in vitro

Potential genotoxic/cytotoxic effects of the epoxiconazole/fenpropimorph-based fungicide were investigated using single cell gel electrophoresis and cytogenetic assays: chromosomal aberrations, sister chromatid exchanges, micronuclei and fluorescence in situ hybridization in cultured bovine lymphocytes. No statistically significant elevations of DNA damage and increases in cytogenetic endpoints were seen. However, evident cytotoxic effect presented as a decrease in mitotic and proliferation indices were recorded after exposure of bovine lymphocytes to the fungicide for 24 and 48 h at concentrations ranging from 3 to 15 µg mL^?1 (P < 0.05, P < 0.01, P < 0.001). Similarly, for 24 h an inhibition in the cytokinesis block proliferation index (CBPI) was obtained after exposure to the fungicide at concentrations ranging from 1.5 to 15 µg mL^?1 (P < 0.01, P < 0.001) in each donor.     

Remediation capacity of Cd and Pb ions by mycelia of Imleria badia,Laetiporus sulphureus,and Agaricus bisporus in vitro cultures

The goal of this study was to evaluate cadmium and lead accumulation ability of in vitro cultures biomass containing selected edible mushroom species derived from the environment (Laetiporus sulphureus, Imleria badia) and those of commercial origin (Agaricus bisporus). Atomic absorption spectrometry was used to evaluate the content of Cd(II) and Pb(II) on the medium supplemented with Cd(II) or Pb(II), each of them at the same concentration of 5·10^?5 M. The highest concentration of Cd(II) ions was determined in the biomass from L. sulphureus in vitro cultures, while the highest concentration of Pb(II) ions was found in the biomass from A. bisporus in vitro cultures. The greatest Cd(II) and Pb(II) accumulation ability in mycelium per dry weight was shown for L. sulphureus. Among the test species, biomass of A. bisporus showed the lowest ability for the bioaccumulation of Cd(II); however, comparable ability for the remediation of Pb(II) was provided by the biomasses from A. bisporus and I. badia in vitro cultures. The results confirm the possibility of using these mushroom species for remediation and indicate the relationship between bioaccumulation of heavy metals and the test species.     

Chronic exposure to contaminated drinking water stimulates PPAR expression in mice livers

Mice were fed with source water (SW) and tap water (TW) for 90 d to evaluate hepatotoxicity induced by the drinking water. Histopathologic observation showed no obvious damage to hepatic tissue in the SW and TW groups. However, microarray analysis indicated that the SW and TW exposures affected many metabolic pathways, among which PPAR (peroxisome proliferator-activated receptors) signaling was most susceptible. Immunohistochemical staining demonstrated that both PPAR-α and PPAR-γ were significantly increased in the exposure groups compared to control. Enzyme-linked immunosorbent assay revealed that PPAR-α expression level was increased from 23.37±0.53 ng g(-1) liver weight in control group to 26.60±1.43 ng g(-1) liver weight in SW group and 27.68±1.10 ng g(-1) liver weight in TW group (p<0.05). For PPAR-γ, the expression level was also significantly enhanced from 0.83±0.07 ng g(-1) liver weight in control group to 1.11±0.20 ng g(-1) liver weight in SW group and 1.16±0.07 ng g(-1) liver weight in TW group (p<0.05). The SW and DW posed no obvious hepatotoxicity on mice and PPAR-α/-γ could be used as a novel biomarker to assess public health risk induced by slightly contaminated drinking water.     

The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes

The potential genotoxic effect of thiacloprid formulation on bovine peripheral lymphocytes was evaluated using the comet assay and the cytogenetic endpoints: chromosome aberrations (CAs), sister chromatid exchanges (SCEs) and micronuclei (MNi). Whole blood cultures were treated with the insecticide at concentrations of 30, 60, 120, 240 and 480 μg mL^?1 for 24, 48 h and/or 2 h of incubation. A statistically significant increase in the frequency of DNA damage, as well as in unstable chromosome aberrations (% breaks) were found after exposure to the insecticide at concentrations ranging from 120 to 480 μg mL^?1 (P < 0.05, P < 0.01, P < 0.001). For the detection of stable structural chromosome aberrations (e.g., translocations) and numerical aberrations by the FISH method, three whole chromosome painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) were used in our experiments. We observed numerical aberrations, but without any statistical significance. Regarding the sister chromatid exchanges, no significant elevation in the SCE frequencies was found after 24-h exposure to the insecticide. A dose-related response in the SCE induction was obtained in bovine cultures after the prolonged time of exposure (48 h) to thiacloprid formulation at concentrations ranging from 120 to 480 μg mL^?1 in each donor (P < 0.05, P < 0.01), which was associated with a reduction of the PI (P < 0.05, P < 0.01). The insecticide failed to produce MNi; however, a significant reduction of CBPI was observed. Using real-time PCR, a decrease in the expression of bovine glutathione S-transferase M3 (GSTM3) was detected at the lowest dose. The higher concentrations of thiacloprid formulation caused an increase in the mRNA expression.     

Metallothionein induction, antioxidative responses, glycogen and growth changes in Tubifex tubifex (Oligochaete) exposed to the fungicide, fenhexamid

Laboratory studies were conducted to determine the effects of different concentrations of fenhexamid (0.1, 1, and 10 mg L(-1)) on growth, oxidative stress, protein, glycogen, and metallothionein (MT) contents in Tubifex tubifex after an exposure of 2, 4, and 7 days. In addition, residues of the fungicide were followed in water and in the worms. In water, fenhexamid concentration decreased slowly (maximum -2 +/- 0.03% after 2 days for 1 mg L(-1)). In the worms, it increased after 4 days and decreased thereafter, confirming that the worms were exposed to the fungicide and not to a degradation product. LC50 values were between 95.22 +/- 5.36 and 32.11 +/- 1.8 mg L(-1) depending on exposure time. Exposure to fenhexamid had a negative effect on T. tubifex growth (maximum effect -12.2 +/- 0.8% after 7 days with 10 mg L(-1)) demonstrating the toxic effect of the pesticide. This growth rate decrease was accompanied by a reduction in protein and glycogen contents. The activity of catalase (CAT), and glutathione reductase (GR) increased in response to the fungicide demonstrating an oxidative stress in the worms. In contrast glutathion-S-transferase activity (GST) decreased. Exposure to fenhexamid also induced synthesis of MT (maximum +78 +/- 8% after 2 days for 10 mg L(-1)). The specificity of MT concentration increase in response to metals is discussed.     

Toxicity and removal of pesticides by selected aquatic plants

Pesticides are being detected in water bodies on an increasingly frequent basis. The present study focused on the phytoremediation potential of selected aquatic plants to remove phytosanitary products from contaminated water. We investigated the uptake capacity of Lemna minor (L. minor), Elodea canadensis (E. canadensis) and Cabomba aquatica (C. aquatica) on three pesticides: copper sulphate (fungicide), flazasulfuron (herbicide) and dimethomorph (fungicide). Pesticide toxicity was evaluated by exposing plants to five concentrations (0-1 mg L(-1)) in culture media for 7d using chlorophyll fluorescence as a biomarker. The toxicity of the contaminants was the same for all the aquatic plants studied and occurred in this descending order of toxicity: flazasulfuron>copper>dimethomorph. We found that L. minor had the most efficient uptake capacity, followed by E. canadensis and then C. aquatica. The maximum removal rate (microg g(-1)fresh weight d(-1)) of copper, flazasulfuron and dimethomorph was 30, 27 and 11, respectively.     

Antioxidant responses and reactive oxygen species generation in different body regions of the estuarine polychaeta Laeonereis acuta (Nereididae)

The aim of this study was to analyze the total antioxidant capacity (TOSC), generation of reactive oxygen species (ROS) and lipid peroxidation (LPO) in the different body regions of the estuarine polychaeta Laeonereis acuta (Nereididae) sampled at non-polluted (NOPOL) and polluted (POL) sites from Lagoa dos Patos (Southern Brazil). Organisms collected at POL during summer showed similar (p>0.05) TOSC values along the body, but worms collected at NOPOL presented higher (p<0.05) TOSC values in the posterior (P) region in respect of anterior (A) region and middle (M) region. TOSC in the P region at NOPOL was higher (p<0.05) compared with the same body region of worms at POL. In summer, ROS concentration was higher in A and M regions of worms at POL in respect of the organisms at NOPOL. During winter all the regions showed higher ROS in worms sampled at POL. It was registered absence of season influence on LPO content, but in the P region at NOPOL in summer there were lower LPO levels compared with the others regions (p<0.05). In vitro assays showed that P region, despite a higher basal ROS, presented a higher competence to cope with pro-oxidants compared with A and M regions (p<0.05), corroborating the field results. A lower proteic sulfhydril content was observed in P in respect of the other regions (p<0.05) supporting the idea of a highest oxidant condition in this region. The results indicate that worms collected at the POL site are confronted to higher ROS concentrations, affecting its antioxidant capacity, a result that depends of body regions.     

Enzyme and fungal treatments and a combination thereof reduce olive mill wastewater phytotoxicity on Zea mays L. seeds

The phytotoxicity of olive-mill wastewater (OMW) has been suggested to be mainly due to its phenolic components. This study investigated the impact of three different low-cost dephenolization treatments on the wastewater phytotoxicity. To this aim, germinability of maize (Zea mays L.) seeds sown on a sandy-loamy soil which had been spread with different volumes (from 40 to 160m(3)ha(-1)) of either biologically-treated OMW or relative incubation control was determined. Biological treatments included either Panus tigrinus liquid cultures or incubation with commercial laccase (1UIml(-1)) or an innovative sequential combination of laccase and P. tigrinus cultures. All treatments markedly reduced phytotoxicity and promising results were obtained with commercial laccase. In fact, germinability and mean germination times in soil spread with laccase-treated OMW, did not significantly differ from those observed in soil irrigated with tap water (control) up to OMW volumes of 120m(3)ha(-1). Although the highest phenol reduction (ca. 81%) was obtained by the sequential use of laccase and P. tigrinus, the feasibility of the enzyme treatment is undoubtedly more convincing under the technological point of view.     

Validation and global uncertainty of a gas chromatographic with mass spectrometry method for fenamidone analysis in grapes and wines

Fenamidone is an imidazolinone fungicide recently introduced in viticulture practices. This work reports the validation and assessment of global uncertainty of a gas chromatographic with mass spectrometry method to analyze fenamidone in grapes and wines. This method consists in a simple and fast liquid-liquid extraction step followed by chromatographic determination. Limits of detection for fenamidone in grapes and wines were, respectively, 0.05 mg/kg and 0.06 mg/L, precision was below 9.4% and average recovery was 89 +/- 5%. In the concentration range from 0.05 to 1.00 mg/kg (or mg/L) of fenamidone, global uncertainty calculated following the EURACHEM/CITAC rules, and also by the Horwitz function, was below 25%. The EURACHEM/CITAC global uncertainty budget used gave lower estimates than those obtained from the Horwitz function.     

Effects of feed-supplementation and hide-spray application of two sources of tannins on enteric and hide bacteria of feedlot cattle

Pathogenic bacteria attached to the hide or shed in the feces of cattle at slaughter can contaminate carcasses intended to be processed for human consumption. Therefore, new pre-harvest interventions are needed to prevent the carriage and excretion of foodborne pathogens in cattle presented to the processing plant. The objectives of this study were to examine the antimicrobial effects of hydrolysable tannin-rich chestnut and condensed tannin-rich mimosa extracts on bacterial indicators of foodborne pathogens when applied as a hide-intervention and as a feed additive to feedlot cattle. Water (control) or solutions (3 % wt/vol) of chestnut- and mimosa-extract treatments were sprayed (25 mL) at the left costal side of each animal to a 1000 cm2 area, divided in four equal quadrants. Hide-swabs samples obtained at pre-, 2-min, 8-h, and 24-h post-spray application were cultured to enumerate Escherichia coli/total coliforms and total aerobic plate counts. In a second experiment, diets supplemented without (controls) or with (1.5 % of diet dry matter) chestnut- or mimosa-extracts were fed during a 42-day experimental feeding period. Weekly fecal samples starting on day 0, and rumen fluid obtained on days 0, 7, 21 or 42 were cultured to enumerate E.coli/total coliforms and Campylobacter. Tannin spray application showed no effect of treatment or post-application-time (P > 0.05) on measured bacterial populations, averaging 1.7/1.8, 1.5/1.6 and 1.5/1.7 (log??CFU/cm2) for E. coli/total coliforms, and 4.0, 3.4 and 4.2 (log??CFU/cm2) in total aerobes for control, chestnut and mimosa treatments, respectively. Mean (± SEM) ruminal E. coli and total coliform concentrations (log(10) CFU/mL) were reduced (P < 0.01) in steers fed chestnut-tannins (3.6 and 3.8 ± 0.1) in comparison with the controls (4.1 and 4.2 ± 0.1). Fecal E. coli concentrations were affected by treatment (P< 0.01), showing the highest values (log?? CFU/g) in fecal contents from mimosa-fed steers compared to controls (5.9 versus 5.6 ± 0.1 SEM, respectively). Total coliforms (log CFU/g) showed the highest values (P < 0.01) in feces from chestnut- and mimosa-fed steers (6.0 and 6.1 ± 0.1 respectively) in comparison with controls (5.7 ± 0.1). Fecal Campylobacter concentrations (log??CFU/g) were affected by treatment (P < 0.05), day (P < 0.001) and their interaction (P < 0.01) with the controls having lower concentrations than chestnut- and mimosa-fed steers (0.4, 1.0, and 0.8 ± 0.3, respectively). It was concluded that under our research conditions, tannins were not effective in decreasing measured bacterial populations on beef cattle hides. Additionally, chestnut tannin reduced E. coli and total coliforms within the rumen but the antimicrobial effect was not maintained in the lower gastrointestinal tract. Further research is necessary to elucidate the possible antimicrobial effects of tannins at site-specific locations of the gastrointestinal tract in beef cattle fed high-grain and high-forage diets.     

Human emotional stress, dioxin blood content and genetic damage in Chapaevsk town

Forty five women from Chapaevsk town (three groups of 14-16 persons aged 20-40 yr) with different exposures to dioxins and different dioxin levels in blood were tested by four standard psychological questionnaires to detect emotional stress. Results of psychological testing were compared with individual dioxin blood contents and chromosome aberration level in blood cells. Results of the investigation were the next: (1) three groups tested differed significantly (P < or = 0.05) in emotional stress level, thus that highest level of stress was detected within the group with the highest dioxin blood contents (group "Workers"); (2) high level of correlation between emotional stress and individual dioxins blood contents (up P < or = 0.001) as well as between emotional stress and individual chromosome aberration level (up P < or = 0.05) was revealed. These results are discussed from the position of desadaptation induced by dioxins, which led to chromosome aberration induction.     

Pesticide levels in surface waters in an agricultural-forestry basin in Southern Chile

Residues of five pesticides in surface water were surveyed during 2001 and 2003 in the Traiguen river basin in Southern Chile. Simazine, hexazinone, 2,4-D, picloram herbicides and carbendazim fungicide were selected through a pesticide risk classification index. Six sampling stations along the river were set up based on agricultural and forestry land use. The water sampling was carried out before and after the pesticide application periods and in correspondence to some rain events. Pesticides were analyzed by HPLC with DAD detection in a multiresidue analysis. During 2001, in the first sampling campaign (March), the highest concentrations of pesticides were 3.0 microg l(-1) for simazine and hexazinone and 1.8 microg l(-1) for carbendazim. In the second sampling (September), the highest concentration were 9.7 microg l(-1) for 2,4-D, 0.3 microg l(-1) for picloram and 0.4 microg l(-1) for carbendazim. In the last sampling period (December), samples indicated contamination with carbendazim fungicide at levels of up to 1.2 microg l(-1). In sampling carried out on May 2003, no pesticides were detected. In October 2003, the highest concentrations of pesticides were 4.5 microg l(-1) for carbendazim and 2.9 microg l(-1) for 2,4-D. Data are discussed in function of land use and application periods of the products, showing a clear seasonal pattern pollution in the Traiguen river. Risk assessment for these pesticides was calculated by using a risk quotient (RQ = PNEC/PEC). For picloram the calculated RQ < was 0, which indicates that no adverse effects may occur due to the exposure to this herbicide in the Traiguen river basin. For 2,4-D, simazine, hexazinone, carbendazim RQ > 1, meaning that adverse effects could occur and it is necessary to reduce pesticide exposure in surface waters. It is recommended to continue with a pesticide monitoring program and the implementation of ecotoxicological testing with local and standardized species in order to consider the probability of effects occurrence, with less uncertainty. Thus, it will be more feasible to make some recommendations to regulatory agencies regarding the pesticide use.     

Determination of wine microbiota using classical method, polymerase chain method and Step One Real-Time PCR during fermentation process

The aim of our study was the identification of grape, must and wine microbiota during the fermentation process using a classical microbiological method and Real-Time PCR. The changes in different groups of microorganisms were monitored in total counts of bacteria, lactobacilli and yeasts. Microbiological parameters were observed during the current collection and processing of grapes in 2009. Samples were taken during the fermentation process in wine enterprises and a private vineyard. During this period 30 samples of wine among Müller Thurgau, Cabernet Sauvignon, Chardonnay, Tramin and Red Bio-wine were examined. Samples were collected from stages of grape-must unfiltered, grape-must filtered, the beginning of fermentation, fermentation, late fermentation and young wine. The highest total counts of bacteria ranged from 0.00 to 176 ± 15 CFU.mL(-1) in the wine of Müller Thurgau, the highest number of yeast ranged from 0.00 to 150 ± 9 CFU.mL(-1) in the wine of Müller Thurgau and the number of Lactobacillus spp. ranged from 0.00 to 92 ± 5 CFU.mL(-1) in the sample of Cabernet Sauvignon wine. The presence and sensitivity of Gram-positive and Gram-negative bacterial species Enterococcus faecium, Lactobacillus acidophilus, Lactobacillus crispatus and Lactobacillus salivarius were detected using Real-Time PCR (RTQ PCR). Susceptibility of Enterococcus faecium varied in different isolates from 1 to 10(6) CFU.mL(-1), the sensitivity of the species Lactobacillus acidophilus in different isolates of the wine samples ranged from 1 to 10(5) CFU.mL(-1). We also monitored representation of species Lactobacillus crispatus, which were captured by RTQ PCR sensitivity and ranged from 1 to 10(5) CFU.mL(-1). Identification of the species Lactobacillus salivarius in each of isolates by RTQ PCR method showed the presence of these bacteria in the range of 1 to 10(4) CFU.mL(-1).     

Microbial degradation of mefenoxam in rhizosphere of Zinnia angustifolia

The fate of the fungicide mefenoxam was studied in a containerized rhizosphere system. The rhizosphere system used Zinnia angustifolia (Tropic Snow) in a bark/sand potting mix and was compared to bulk potting mix (no plants). Rhizosphere microbial populations were allowed to establish for 3 weeks prior to fungicide addition (20 microg per g mix). Mefenoxam and degradation product concentrations were determined by High HPLC or capillary electrophoresis after extraction. Seventy eight percent of the fungicide originally applied to the rhizosphere was degraded after 21 days compared to 44% in bulk system (no plant). The primary degradation product was the free acid N-(2,6-dimethylphenyl)-N-(methoxyacetyl)-DL-alanine, which accounted for 71% of the applied parent chemical after 30 days. N-(2,6-dimethylphenyl)-acetamide was also detected, but in lesser amounts. Bacterial populations in the rhizosphere increased during the 30-day period, which correlated with an increase in degradation of the parent compound. Pure cultures of Pseudomonas fluorescens and Chrysobacterium indologenes isolated from the rhizosphere system could degrade the applied fungicide (10 microg/ml) almost completely to the free acid within 54 h.     

Validation and global uncertainty of a gas chromatographic with mass spectrometry method for fenamidone analysis in grapes and wines

Fenamidone is an imidazolinone fungicide recently introduced in viticulture practices. This work reports the validation and assessment of global uncertainty of a gas chromatographic with mass spectrometry method to analyze fenamidone in grapes and wines. This method consists in a simple and fast liquid-liquid extraction step followed by chromatographic determination. Limits of detection for fenamidone in grapes and wines were, respectively, 0.05 mg/kg and 0.06 mg/L, precision was below 9.4% and average recovery was 89 ± 5%. In the concentration range from 0.05 to 1.00 mg/kg (or mg/L) of fenamidone, global uncertainty calculated following the EURACHEM/CITAC rules, and also by the Horwitz function, was below 25%. The EURACHEM/CITAC global uncertainty budget used gave lower estimates than those obtained from the Horwitz function.     

Arsenic accumulation in duckweed (Spirodela polyrhiza L.): a good option for phytoremediation

Some unavoidable drawbacks of traditional technologies have made phytoremediation a promising alternative for removal of arsenic from contaminated soil and water. In the present study, the potential of an aquatic macrophyte Spirodela polyrhiza L. for phytofiltration of arsenic, and the mechanism of the arsenic uptake were investigated. The S. polyrhiza L. were grown in three test concentrations of arsenate and dimethylarsinic acid (DMAA) (i.e. 1.0, 2.0 and 4.0microM) with 0 (control), 100 or 500microM of phosphate. One control treatment was also set for each test concentrations of arsenic. The PO(4)(3-) concentration in control treatment was 0.02microM. When S. polyrhiza L. was cultivated hydroponically for 6d in culture solution containing 0.02microM phosphate and 4.0microM arsenate or DMAA, the arsenic uptake was 0.353+/-0.003micromolg(-1) and 7.65+/-0.27nmolg(-1), respectively. Arsenic uptake into S. polyrhiza L. was negatively (p<0.05) correlated with phosphate uptake when arsenate was applied to the culture solutions owing to similar in the sorption mechanism between AsO(4)(3-) and PO(4)(3-), and positively (p<0.05) correlated with iron uptake due to adsorption of AsO(4)(3-) onto iron oxides. Thus, the S. polyrhiza L. accumulates arsenic by physico-chemical adsorption and via the phosphate uptake pathway when arsenate was added to the solutions. These results indicate that S. polyrhiza L. would be a good arsenic phytofiltrator. In contrast, DMAA accumulation into S. polyrhiza L. was neither affected by the phosphate concentration in the culture nor correlated (p>0.05) with iron accumulation in plant tissues, which indicates that S. polyrhiza L. uses different mechanisms for DMAA uptake.