Phytotoxicity of nanoparticles—problems with bioassay choosing and sample preparation

For a full estimation of the risk related with the presence of engineered nanoparticles (ENPs) in the environment, the use of the current ecotoxicological methods may prove insufficient. In the study presented herein, various methods of assessment of ecotoxicity were applied to compare the phytotoxicity of three ENPs: nano-ZnO, nano-TiO₂ and nano-Ni. The toxicity was assayed both for aqueous solutions of the ENPs (the germination/elongation test and Phytotestkit F^TM) and for ENPs added to soil (Phytotoxkit F^TM and modified Phytotoxkit F^TM). Lepidium sativum was used as a test plant. The scope of the study also included the assessment of the effect of the method of ENP application to the soil (as powder and aqueous suspension) on their phytotoxicity. In the course of the study, no effect of the studied ENPs and their bulk counterparts on the germination of seeds was observed. The root growth inhibition of L. sativum depended on the kind of test applied. The trend between concentration of ENPs and effect depended on the method used and kind of ENPs. For most nanoparticles (despite of the method used), the differences in phytotoxicity between nano and bulk particles were observed. Depending on the kind of ENPs, their phytotoxicity differs between water and soil. ZnO (nano and bulk) and nano-Ni were more toxic in soil than in water. For TiO₂ and bulk-Ni, reverse trend was observed. A different method of ENP application to soil differently affects the phytotoxicity.     

Haematology, genotoxicity, enzymatic activity and histopathology as biomarkers of metal pollution in the shrew Crocidura russula

Haematological (WBC, RBC, Hgb and Hct) and genotoxicity (MNT) parameters, hepatic enzymatic activities (GST, GPx and GR), and a histopathological evaluation of liver, kidneys and gonads were assessed as general biomarkers of metal pollution in the shrew Crocidura russula inhabiting a pyrite mining area. Specimens exposed to metals presented a few significant alterations when compared with reference animals: GST activity decreased; micronuclei increased; and evident liver alterations related to metal exposure were observed. On the basis of all the parameters studied, age was an important factor that partly explained the observed variation, whereas sex was the least important factor. Significant correlations were also found between heavy metal concentrations and biomarkers evaluated, demonstrating the great influence of these metals in the metabolic alterations. To the best of our knowledge, these data constitute the first measurements of a battery of biomarkers in shrews from a mine site and are among the few available for insectivorous mammals.     

The impact of mandatory versus voluntary participation in the Alberta ignition interlock program

Research has demonstrated that participation in an interlock program significantly reduces the likelihood of subsequent driving while intoxicated (DWI) convictions at least so long as the interlock device is installed in the vehicle. Despite the growing number of jurisdictions that allow interlock programs and the demonstrated success of these programs, the proportion of DWI offenders who actually have the device installed is minimal. In an effort to increase the proportion of offenders using interlocks, some jurisdictions require offenders to install an interlock as a condition of license reinstatement whereas others merely offer offenders a reduction in the period of hard suspension if they voluntarily participate in an interlock program. The objective of the present study was to determine the extent to which voluntary interlock participants are more or less successful in terms of subsequent recidivism than those for whom interlock program participation has been mandated. The issue was addressed using data from the interlock program in Alberta, Canada, which provides for both mandatory and voluntary participation. The recidivism experience of voluntary and mandatory interlock participants was examined both during and after the period of interlock installation. Cox regression revealed that, after controlling for (or equating) the number of prior DWI offenses, the survival rates of DWI offenders who were ordered to participate in the interlock program did not differ from those of voluntary participants. These results suggest that further use of mandatory interlock programs should be just as successful as voluntary programs when offenders share characteristics with those studied in Alberta.     

The alcohol interlock: an underutilized resource for predicting and controlling drunk drivers

This report summarizes evidence presented during the Third Annual Ignition Interlock Symposium at Vero Beach, Florida, 29 October 2002. The ignition interlock prevents a car from starting when blood alcohol concentration (BAC) is elevated. We review some of our prior work as well as introduce previously unpublished results to demonstrate the manner in which the data recorded by the alcohol ignition interlock device can serve as an advance predictor of future driving under the influence (DUI) of alcohol risks. Data used in this current report represent approximately 2,200 ignition interlock users from Alberta, Canada, and about 8,000 interlock users from Quebec, Canada; the Alberta data set contained 5.5 million breath tests and the Quebec data 18.8 million breath tests. All tests are time and date stamped and this information was used to characterize patterns of BAC and vehicle use, and the relationship between BAC elevations and DUI offenses that accumulated after the interlock was removed from the vehicles. Findings from Cox regression (Marques et al., 2003) show that BAC elevations > .02-.04% are more potent predictors of repeat DUI (p < .0001) than even prior DUI (p < .006), usually found to be the strongest indicator of driver risk. Prior DUI obviously has no use for scaling the risk of first-time offenders. Drivers who are both multiple offenders and who have more than a few elevated interlock BAC tests are much more likely to repeat DUI. The timing and pattern of elevated BAC tests provided during the time drivers were required to use an alcohol ignition interlock device are remarkably similar on both a daily basis and an hourly basis when the interlock programs from the two provinces are compared directly. Both provinces had higher rates of elevated tests on Saturday and Sunday, and the fewest elevated tests on Tuesdays. The absolute rate of elevated tests is similar despite the two provinces adhering to different interlock lockout points (.02% Quebec; .04% Alberta). Charts tracking the Monday-Friday timing of elevated BAC tests by hour are nearly identical for both provinces. The most elevated BAC tests occurred between 7 and 9 A.M. Monday to Friday, even though most vehicle start attempts occurred much later in the day. This higher rate of elevated morning BAC likely represents drinking from the prior evening with alcohol not yet cleared from circulation; those with elevated BAC in the early morning were more likely to have a repeat offense even after accounting for prior DUI and the higher overall rate of elevated BAC tests. This is viewed as evidence of a drinking problem that will lead to impaired driving after the controlling function of the interlock is removed. Policy changes are discussed that might take better advantage of interlock information to improve the public response to drunk driving.     

The alcohol interlock: an underutilized resource for predicting and controlling drunk drivers

This report summarizes evidence presented during the Third Annual Ignition Interlock Symposium at Vero Beach, Florida, 29 October 2002. The ignition interlock prevents a car from starting when blood alcohol concentration (BAC) is elevated. We review some of our prior work as well as introduce previously unpublished results to demonstrate the manner in which the data recorded by the alcohol ignition interlock device can serve as an advance predictor of future driving under the influence (DUI) of alcohol risks. Data used in this current report represent approximately 2,200 ignition interlock users from Alberta, Canada, and about 8,000 interlock users from Quebec, Canada; the Alberta data set contained 5.5 million breath tests and the Quebec data 18.8 million breath tests. All tests are time and date stamped and this information was used to characterize patterns of BAC and vehicle use, and the relationship between BAC elevations and DUI offenses that accumulated after the interlock was removed from the vehicles. Findings from Cox regression show that BAC elevations >.02-.04% are more potent predictors of repeat DUI (p<.0001) than even prior DUI (p<.006), usually found to be the strongest indicator of driver risk. Prior DUI obviously has no use for scaling the risk of first-time offenders. Drivers who are both multiple offenders and who have more than a few elevated interlock BAC tests are much more likely to repeat DUI. The timing and pattern of elevated BAC tests provided during the time drivers were required to use an alcohol ignition interlock device are remarkably similar on both a daily basis and an hourly basis when the interlock programs from the two provinces are compared directly. Both provinces had higher rates of elevated tests on Saturday and Sunday, and the fewest elevated tests on Tuesdays. The absolute rate of elevated tests is similar despite the two provinces adhering to different interlock lockout points (.02% Quebec;.04% Alberta). Charts tracking the Monday-Friday timing of elevated BAC tests by hour are nearly identical for both provinces. The most elevated BAC tests occurred between 7 and 9 A.M. Monday to Friday, even though most vehicle start attempts occurred much later in the day. This higher rate of elevated morning BAC likely represents drinking from the prior evening with alcohol not yet cleared from circulation; those with elevated BAC in the early morning were more likely to have a repeat offense even after accounting for prior DUI and the higher overall rate of elevated BAC tests. This is viewed as evidence of a drinking problem that will lead to impaired driving after the controlling function of the interlock is removed. Policy changes are discussed that might take better advantage of interlock information to improve the public response to drunk driving.     

From Environmental Ethics to Nature Conservation Policy: Natura 2000 and the Burden of Proof

Natura 2000 is a network of natural sites whose aim is to preserve species and habitats of relevance in the European Union. The policy underlying Natura 2000 has faced widespread opposition from land users and received extensive support from environmentalists. This paper addresses the ethical framework for Natura 2000 and the probable moral assumptions of its main stakeholders. Arguments for and against Natura 2000 were analyzed and classified according to “strong” or “weak” versions of the three main theories of environmental ethics – anthropocentrism, biocentrism, and ecocentrism. Weak (intergenerational) anthropocentrism was found to underlie the Natura 2000 network itself and the positions of environmentalists, while strong (traditional) anthropocentrism pervaded the positions of economic developers. Land users seemed to fall somewhere between weak and strong anthropocentrism. The paper discusses the relation between ethics and different attitudes towards Natura 2000, highlighting some of the implications for the network’s ongoing implementation. It is shown that Natura 2000 achieves a strong reversal of the burden of proof from conservation to economic development and land use change under anthropocentrism. It is argued that the alleged theoretical divide between anthropocentrism and non-anthropocentrism in relation to the burden of proof does not seem to hold in practice. Finally, it is predicted that the weak versions of anthropocentrism, biocentrism, and ecocentrism, are likely to converge extensively in respect to nature conservation policy measures.“Charting the depths of law and policy disputes yield an understanding of ethical differences.” Paul B. Thompson (2002, p. 189).     

Metallothionein levels in Algerian mice (Mus spretus) exposed to elemental pollution: an ecophysiological approach

The potential use of metallothioneins (MTs) as biomarkers of trace metal contamination was evaluated for the first time in the Algerian mouse (Mus spretus). Mice were collected seasonally in an abandoned mining area (Aljustrel) and in a reference area, both located in southern Portugal. MT levels were quantified in liver and kidney by differential pulse polarography and hepatic elemental concentrations (Mn, Fe, Cu, Zn, Se) were determined by particle-induced X-ray emission. Hepatic iron and selenium concentrations were elevated in mice from Aljustrel mine when compared to reference animals. MTs levels were averagely higher in mice from Aljustrel than those originated from the reference area. A season-dependent significant effect was found on the hepatic and renal MT concentrations, characterized by higher levels in winter and lower in autumn. In contaminated mice positive relationship between liver elemental contents (Cu in autumn and Fe in winter) and MTs were found. The seasonal variation of MT suggests that probably physiological and environmental factors could influence hepatic and renal MT induction. Results seem to imply that some environmental disturbance occur in the vicinity of the Aljustrel mine. Therefore, for the management purposes MT levels should be followed in liver of M. spretus, especially in winter. Furthermore, other physiological factors that could influence MT expression and turnover in Algerian mouse should also be monitored.     

Application of manure and compost to contaminated soils and its effect on zinc accumulation by Solanum nigrum inoculated with arbuscular mycorrhizal fungi

Zn accumulation in Solanum nigrum grown in naturally contaminated soil in the presence of different types of organic amendments was assessed. Under the same conditions, the response of the plant to inoculation with two different isolates of arbuscular mycorrhizal fungi (AMF) (Glomus claroideum and Glomus intraradices) was also evaluated. S. nigrum grown in the non-amended soil always presented higher Zn accumulation in the tissues, with the addition of amendments inducing reductions of up to 80 and 40%, for manure and compost, respectively, and enhancing plant biomass yields. The establishment of S. nigrum in the Zn contaminated soil combined with the application of amendments led to a 70-80% reduction in the amount of Zn leached through the soil. The use of S. nigrum in combination with manure appeared as an effective method for reducing the effects of soil contamination, diminishing Zn transfer to other environmental compartments via percolation.     

EDDS and EDTA-enhanced zinc accumulation by Solanum nigrum inoculated with arbuscular mycorrhizal fungi grown in contaminated soil

The effect of two different chelating agents [EDTA and EDDS (S,S-ethylenediaminedissucinic acid)] on Zn tissue accumulation in Solanum nigrum L. grown in a naturally contaminated soil was assessed. Under those conditions, the response of the plant to the inoculation with two different isolates of arbuscular mycorrhizal fungi (AMF)--Glomus claroideum and Glomus intraradices--was also studied. Plants grown in the local contaminated soil (Zn levels of 433mg kg(-1)) accumulated up to 1191mg kg(-1) of Zn in the roots, 3747mg kg(-1) in the stems and 3409mg kg(-1) in the leaves. S. nigrum plants grown in the same soil spiked with extra Zn (Zn levels of 964mg kg(-1)) accumulated up to 4735, 8267 and 7948mg Zn kg(-1) in the leaves, stems and roots, respectively. The addition of EDTA promoted an increase in the concentration of Zn accumulated by S. nigrum of up to 231% in the leaves, 93% in the stems and 81% in the roots, while EDDS application enhanced the accumulation in leaves, stems and roots up to 140, 124 and 104%, respectively. In the stems, the presence of Zn was predominantly detected in the cortex collenchyma cells, the starch sheath and the internal phloem and xylem parenchyma, and the addition of chelating agents did not seem to have an effect on the localisation of accumulation sites. The devise of a chelate-enhanced phytoextraction strategy, using chelating agents and AMF, is discussed.     

The use of the oxidative stress responses as biomarkers in Nile tilapia (Oreochromis niloticus) exposed to in vivo cadmium contamination

Water contaminants have a high potential risk for the health of populations. Protection from toxic effects of environmental water pollutants primarily involves considering the mechanism of low level toxicity and likely biological effects in organisms who live in these polluted waters. The biomarkers assessment of oxidative stress and metabolic alterations to cadmium exposure were evaluated in Nile tilapia, Oreochromis niloticus. The fish were exposed to 0.35, 0.75, 1.5, and 3.0 mg/l concentrations of Cd2+ (CdCl2) in water for 60 days. Fish that survived cadmium exposure showed a metabolic shift and a compensatory development for maintenance of the body weight gain. We observed a decreased glycogen content and decreased glucose uptake in white muscle. Lactate dehydrogenase (LDH) and creatine phosphokinase (CK) activities were also decreased, indicating that the glycolytic capacity was decreased in this tissue. No alterations were observed in total protein content in white muscle due to cadmium exposure suggesting a metabolic shift of carbohydrate metabolism to maintenance of the muscle protein reserve. There was an increase in glucose uptake, CK increased activity, and a clear increase of LDH activity in red muscle of fish with cadmium exposure. Since no alterations were observed in lipoperoxide concentration, while antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were changed in the liver and the red and white muscle of fish with cadmium exposure, we can conclude that oxygen free radicals are produced as a mediator of cadmium toxicity. Resistance development is related with increased activities of antioxidant enzymes, which were important in the protection against cadmium damage, inhibiting lipoperoxide formation.