Atrazine induction of cytochrome P450 in Chironomus tentans larvae

Cytochrome P450-dependent aldrin epoxidation was characterized in third instar larvae of the aquatic midge, Chironomus tentans. Optimal in vitro assay conditions for the epoxidase were pH 7.6 and 31 degrees C. Activity was linear up to 40 min of incubation time and 0.5 mg microsomal protein per incubation. The activity was concentrated in the microsomal fraction of whole body homogenates and was NADPH-dependent. The effect of atrazine exposure on aldrin epoxidase was measured to determine if this herbicide induces cytochrome P450-dependent activity. Comparisons of control and atrazine-exposed midges indicated increased epoxidase activity as a result of atrazine exposure, and a 45 kDa protein of increased intensity was observed after SDS-PAGE of microsomal protein. The molecular weight of this protein was similar in size to cytochrome P450 enzymes reported for other insects. Heme staining of SDS-PAGE gels and immunochemical studies using a Drosophila melanogaster anti-P450 polyclonal antiserum, further support the cytochrome P450 nature of this inducible 45 kDa protein.     

Soil organic matter mobilization by root exudates

In order to study the different soil organic matter mobilisation by agrarian (Zea mais: cultivars Paolo and Sandek) and forest (Picea abies Karst. and Pinus sylvestris L.) root exudates, three different soils (Dystric Spodic Cambisol--S1, Haplic Luvisol--S2 and Calcaric Cambisol--S3) have been considered. Treating the soils with water (control) or plant root exudates, soil organic matter extracts were obtained. The extracts were characterised by hormone-like activities and gas chromatographic/mass spectrometric (GC/MS) measurements. Water extract and plant root exudates exhibited no hormone-like activity, while the other soil-extracts were endowed with a different hormone-like behaviour. GC/MS data indicated that in the acid soils (S1) Sandek and Picea abies exudates showed a greater ability in extracting organic acid isomers (Cl4COOH, Cl5COOH and Cl7COOH), while in neutral soils (S3) all the exudates were active in separating organic acids. In intermediate conditions (S2), Picea abies and Pinus sylvestris exudates liberated C15COOH isomers, Paolo C11COOH isomers, while Sandek was not effective. The different role of plant root exudates in mobilising bio-molecules from the bulk of the soil is proposed.     

Application of a purge and trap TDS-GC/MS procedure for the determination of emissions from flame retarded polymers

Emissions of volatile organic compounds (VOCs) from different thermoplastic polymers used in electrotechnical applications were investigated using a purge and trap procedure that involved adsorption on Tenax GR. Results were compared to those for an operating TV set monitored in a test chamber. The analyses were in both cases carried out using thermodesorption gas chromatography with mass spectrometric detection (TDS-GC/MS). Substances identified were monomers, volatile additives, or related compounds. Special attention was given to the detection of halogenated compounds. Their origin was studied using reference samples and synthetic standards.     

Seedling insensitivity to ozone for three conifer species native to Great Smoky Mountains National Park

Field symptoms typical of ozone injury have been observed on several conifer species in Great Smoky Mountains National Park, and tropospheric ozone levels in the Park can be high, suggesting that ozone may be causing growth impairment of these plants. The objective of this research was to test the ozone sensitivity of selected conifer species under controlled exposure conditions. Seedlings of three species of conifers, Table Mountain pine (Pinus pungens), Virginia pine (Pinus virginiana), and eastern hemlock (Tsuga canadensis), were exposed to various levels of ozone in open-top chambers for one to three seasons in Great Smoky Mountains National Park in Tennessee, USA. A combination of episodic profiles (1988) and modified ambient exposure regimes (1989-92) were used. Episodic profiles simulated an average 7-day period from a monitoring station in the Park. Treatments used in 1988 were: charcoal-filtered (CF), 1.0x ambient, 2.0x ambient, and ambient air-no chamber (AA). In 1989 a 1.5x ambient treatment was added, and in 1990, additional chambers were made available, allowing a 0.5x ambient treatment to be added. Height, diameter, and foliar injury were measured most years. Exposures were 3 years for Table Mountain pine (1988-90), 3 years for hemlock (1989-91), and 1 and 2 years for three different sets of Virginia pine (1990, 1990-91, and 1992). There were no significant (p<0.05) effects of ozone on any biomass fraction for any of the species, except for older needles in Table Mountain and Virginia pine, which decreased with ozone exposure. There were also no changes in biomass allocation patterns among species due to ozone exposure, except for Virginia pine in 1990, which showed an increase in the root:shoot ratio. There was foliar injury (chlorotic mottling) in the higher two treatments (1.0x and 2.0x for Table Mountain and 2.0x for Virginia pine), but high plant-to-plant variability obscured formal statistical significance in many cases. We conclude, at least for growth in the short-term, that seedlings of these three conifer species are insensitive to ambient and elevated levels of ozone, and that current levels of ozone in the Park are probably having minimal impacts on these particular species.     

Monitoring, modelling and environmental exposure assessment of industrial chemicals in the aquatic environment

Monitoring and laboratory data play integral roles alongside fate and exposure models in comprehensive risk assessments. The principle in the European Union Technical Guidance Documents for risk assessment is that measured data may take precedence over model results but only after they are judged to be of adequate reliability and to be representative of the particular environmental compartments to which they are applied. In practice, laboratory and field data are used to provide parameters for the models, while monitoring data are used to validate the models' predictions. Thus, comprehensive risk assessments require the integration of laboratory and monitoring data with the model predictions. However, this interplay is often overlooked. Discrepancies between the results of models and monitoring should be investigated in terms of the representativeness of both. Certainly, in the context of the EU risk assessment of existing chemicals, the specific requirements for monitoring data have not been adequately addressed. The resources required for environmental monitoring, both in terms of manpower and equipment, can be very significant. The design of monitoring programmes to optimise the use of resources and the use of models as a cost-effective alternative are increasing in importance. Generic considerations and criteria for the design of new monitoring programmes to generate representative quality data for the aquatic compartment are outlined and the criteria for the use of existing data are discussed. In particular, there is a need to improve the accessibility to data sets, to standardise the data sets, to promote communication and harmonisation of programmes and to incorporate the flexibility to change monitoring protocols to amend the chemicals under investigation in line with changing needs and priorities.     

Determination of T-2 toxin in grain and grain products by HPLC and TLC

The purpose of this study was to investigate the T-2 toxin contaminated grain and grain products consumed especially by Turkish population. The T-2 toxin was detected using the high performance liquid chromatography (HPLC) with UV detector at 208 nm and the identify of T-2 was further confirmed by thin layer chromatography (TLC). The recovery was 91 +/- 4.24% for corn flour fortified with the known amount of T-2 toxin (1 ppm). The detection limits of T-2 toxin for the HPLC and the TLC were 25 ng and 50 ng, respectively. A total of 30 commercially available grain and grain product samples were analyzed. Two corn flour samples were found to contain detectable levels of T-2 toxin at a level of 1.60 ppm and 4.08 ppm.     

In situ monitoring of the mutagenic effects of the gaseous emissions of a solid waste incinerator in metropolitan São Paulo, Brazil, using the Tradescantia stamen-hair assay

The present work was designed to determine the potential genotoxicity at the vicinity of a solid waste incinerator in the metropolitan area of S?o Paulo, using the Tradescantia stamen-hair bioassay. Experiments were carried out between December 1998 and April 1999 in four regions (40 pots of plants per site) selected on the basis of their pollution levels predicted by theoretical modeling of the dispersion of the incinerator's plume. The exposure sites were defined as follows: highest level (incinerator); a high level (museum) located 1.5 km from the emission point; a moderate level (school, at a distance of 3.5 km from the incinerator); and a control (at Jaguariúna countryside). The difference in genotoxicity among the groups was statistically significant (p < 0.001). The frequency of mutations observed in the countryside was significantly lower [2.25 +/- 1.55, mean +/- SD (standard deviation)] than that of the sites close to the incinerator. The frequency of mutations measured at the school (3.70 +/- 1.36) was significantly lower than that measured at both the museum (4.89 +/- 1.12) and the incinerator (5.69 +/- 1.34). In conclusion, we found a positive correlation between the spatial distribution of the emissions of the incinerator located in an urban area and the mutagenic events measured by the Tradescantia stamen-hair assay. The in situ approach employed in this study was simple, efficient, and of low cost. No air or chemical extraction of pollutants was necessary for genotoxicity testing as required by other assays.     

Estimating separately personal exposure to ambient and nonambient particulate matter for epidemiology and risk assessment: why and how

This paper discusses the legal and scientific reasons for separating personal exposure to PM into ambient and nonambient components. It then demonstrates by several examples how well-established models and data typically obtained in exposure field studies can be used to estimate both individual and community average exposure to ambient-generated PM (ambient PM outdoors plus ambient PM that has infiltrated indoors), indoor-generated PM, and personal activity PM. Ambient concentrations are not highly correlated with personal exposure to nonambient PM or total PM but are highly correlated with personal exposure to ambient-generated PM. Therefore, ambient concentrations may be used in epidemiology as an appropriate surrogate for personal exposure to ambient-generated PM. Suggestions are offered as to how exposure to ambient-generated PM may be obtained and used in epidemiology and risk assessment.     

Ascorbic acid reduction of residual active chlorine in potable water prior to halocarboxylate determination

In studies on the formation of disinfection byproducts (DBPs), it is necessary to scavenge residual active (oxidizing) chlorine in order to fix the chlorination byproducts (such as haloethanoates) at a point in time. Such research projects often have distinct needs from requirements for regulatory compliance monitoring. Thus, methods designed for compliance monitoring are not always directly applicable, but must be adapted. This research describes an adaptation of EPA Method 552 in which ascorbic acid treatment is shown to be a satisfactory means for reducing residual oxidizing chlorine, i.e., HOCl, ClO-, and Cl2, prior to determining concentrations of halocarboxylates. Ascorbic acid rapidly reduces oxidizing chlorine compounds, and it has the advantage of producing inorganic halides and dehydroascorbic acid as opposed to halogenated organic molecules as byproducts. In deionized water and a sample of chlorinated tap water, systematic biases relative to strict adherence to Method 552 were precise and could be corrected for using similarly treated standards and analyte-fortified (spiked) samples. This was demonstrated for the quantitation of chloroethanoate, bromoethanoate, 2,2-dichloropropanoate (dalapon), trichloroethanoate, bromochloroethanoate, and bromodichlorocthanoate when extracted, as the acids, into tert-butyl methyl ether (MTBE) and esterified with diazomethane prior to gas chromatography with electron capture detection (GC-ECD). Recoveries for chloroethanoate, bromoethanoate, dalapon, dichloroethanoate, trichloroethanoate, bromochloroethanoate, bromodichloroethanoate, dibromoethanoate, and 2-bromopropanoate at concentrations near the lower limit of detection were acceptable. Ascorbic acid reduction appears to be the best option presently available when there is a need to quench residual oxidants fast in a DBP formation study without generating other halospecies but must be implemented cautiously to ensure no untoward interactions in the matrix.