The rotation of white lupin (Lupinus albus L.) with metal-accumulating plant crops: a strategy to increase the benefits of soil phytoremediation.

Most of the plants employed to remove metals from contaminated soils are annuals and have a seed-to-seed life cycle of a few months, usually over spring and summer. Consequently, for most of the year, fields are not actively cleaned but are completely bare and subject to erosion by water and wind. The objective of this study was to evaluate the benefits of using Lupinus albus as a winter crop in a rotation sequence with a summer crop ideally selected for phytoextraction, such as industrial hemp. Lupin plants were grown in two alkaline soil plots (heavy metal-contaminated and uncontaminated) of approximately 400 m(2) each after the cultivation and harvest of industrial hemp. A smaller-scale parallel pot experiment was also performed to better understand the lupin behavior in increasing concentrations of Cd, Cu, Ni and Zn. White lupin grew well in alkaline conditions, covering the soil during the winter season. In few months plants were approximately 40-50 cm high in both control and contaminated plots. In fields where the bioavailable fraction of metals was low (less than 12%), plants showed a high tolerance to these contaminants. However, their growth was affected in some pot treatments in which the concentrations of assimilable Cu, Zn and Ni were higher, ranging from approximately 40-70% of the total concentrations. The lupin's ability to absorb heavy metals and translocate them to shoots was negligible with respect to the magnitude of contamination, suggesting that this plant is not suitable for extending the period of phytoextraction. However, it is entirely exploitable as green manure, avoiding the application of chemical amendments during phytoremediation. In addition, in polluted fields, white lupin cultivation increased the soil concentration of live bacteria and the bioavailable percentage of metals. On average live bacteria counts per gram of soil were 65×10(6)±18×10(6) and 99×10(6)±22*10(6) before and after cultivation, respectively. The percentages of bioavailable Cu, Pb, Ni, Zn and Cr, which were 5.7±0.7, 5.3±1.7, 1.2±0.1, 12±1.5 and 0.1±0.02%, respectively, before lupin growth, increased to 9.6±1.6, 7±2, 2±0.3, 14±1.5 and 0.1±0.02% after lupin harvest. On the whole, our results indicate that the winter cultivation of white lupin in sequence with a metal-accumulator summer crop can improve the recovery of soil quality during the phytoextraction period. It improves the safety of the area, limiting additional ecological and human health problems, and enhances soil health by avoiding the use of chemical amendments and by increasing the levels of viable microorganisms.

Columba livia as a sentinel species for the assessment of urban air genotoxicity.

This study explored the comet assay as a possible tool to assess genotoxicity in erythrocytes of Columba livia to detect genotoxic effects induced by exposure to urban air pollution. Fieldwork was conducted between June 2004 and June 2005 in the city of Milan, Italy, by sampling pigeons in different areas almost twice a week. Six air contaminants-CO, PM10, NO(2), O(3) (ozone), SO(2), and C6H6-plus polycyclic aromatic hydrocarbons (PAHs) in fine particles, temperature, and ultraviolet index, were considered. Genotoxicity levels, expressed as %DNA migrated, tail moment, and damage index (DI), were always higher in wild pigeons than in pigeons living indoors (controls). Animals exposed to urban air showed significant differences from season to season, and the genotoxic parameters presented the highest values in summer (45.30% +/- 1.40% %DNA migrated, 12.73 +/- 0.80 tail moment, and 22.30 +/- 0.15 x DI x 10(-1)); regression analyses showed a positive relation between DI and O(3) concentrations (P < 0.001). The use of the comet assay DI parameter as a rapid assessment of incipient genotoxic risk by pollution, as measured in C. livia living in urban areas, is also discussed.

The arbuscular mycorrhizal fungus Glomus mosseae induces growth and metal accumulation changes in Cannabis sativa L.

The effect of arbuscular mycorrhiza on heavy metal uptake and translocation was investigated in Cannabis sativa. Hemp was grown in the presence and absence of 100 microg g-1 Cd and Ni and 300 microg g-1 Cr(VI), and inoculated or not with the arbuscular mycorrhizal fungus Glomus mosseae. In our experimental condition, hemp growth was reduced in inoculated plants and the reduction was related to the degree of mycorrhization. The percentage of mycorrhizal colonisation was 42% and 9% in plants grown in non-contaminated and contaminated soil, suggesting a significant negative effect of high metal concentrations on plant infection by G. mosseae. Soil pH, metal bioavailability and plant metal uptake were not influenced by mycorrhization. The organ metal concentrations were not statistically different between inoculated and non-inoculated plants, apart from Ni which concentration was significantly higher in stem and leaf of inoculated plants grown in contaminated soil. The distribution of absorbed metals inside plant was related to the soil heavy metal concentrations: in plant grown in non-contaminated soil the greater part of absorbed Cr and Ni was found in shoots and no significant difference was determined between inoculated and non-inoculated plants. On the contrary, plants grown in artificially contaminated soil accumulated most metal in root organ. In this soil, mycorrhization significantly enhanced the translocation of all the three metals from root to shoot. The possibility to increase metal accumulation in shoot is very interesting for phytoextraction purpose, since most high producing biomass plants, such as non-mycorrhized hemp, retain most heavy metals in roots, limiting their application.

The combined toxic and genotoxic effects of Cd and As to plant bioindicator Trifolium repens L.

This study was undertaken to investigate combined toxic and genotoxic effects of cadmium (Cd) and arsenic (As) on white clover, a pollutant sensitive plant frequently used as environmental bioindicator. Plants were exposed to soil spiked with increasing concentrations of cadmium sulfate (20, 40 and 60 mg Kg-1) or sodium arsenite (5, 10 and 20 mg Kg-1) as well as with their combinations. Metal(loid) bioavailability was assessed after soil contamination, whereas plant growth, metal(loid) concentration in plant organs and DNA damage were measured at the end of plant exposition. Results showed that individual and joint toxicity and genotoxicity were related to the concentration of Cd and As measured in plant organs, and that As concentration was the most relevant variable. Joint effects on plant growth were additive or synergistic, whereas joint genotoxic effects were additive or antagonistic. The interaction between Cd and As occurred at both soil and plant level. In soil the presence of As limited the bioavailability of Cd, whereas the presence of Cd increased the bioavailability of As. Nevertheless only As biovailability determined the amount of As absorbed by plants. The amount of Cd absorbed by plant was not linearly correlated with the fraction of bioavailable Cd in soil suggesting the involvement of additional factors, such as plant uptake mechanisms. These results reveal that the simultaneous presence in soil of Cd and As, although producing an additive or synergistic toxic effect on Trifolium repens L. growth, generates a lower DNA damage.

Ecotoxicological characterisation of a mycoherbicide mixture isolated from the fungus Ascochyta caulina.

BACKGROUND: Recent studies have highlighted the possibility of using a mixture of three fungal toxins produced by Ascochyta caulina as a bioherbicide to control noxious weeds. However, to assess the commercial potential of this new bioherbicide, it is necessary to increase knowledge regarding the environmental behaviour of these toxins, and particularly their effects on non-target organisms. A negative ecotoxicological profile of this potential bioherbicide could eventually hinder its registration. RESULTS: In this study, acute and chronic effects of the A. caulina toxins (ACTs) on non-target organisms were evaluated in the laboratory. The (96 h) calculated algal Er C50 and NOEC were 142.7 and <12.5 mg L(-1) respectively. For daphnids, the EC50 (48 h, acute test) was 20.1 mg L(-1) , whereas the NOEC (21 day chronic reproduction test) was 6.3 µg L(-1) . No cases of mortality, sublethal effects or abnormalities were observed in the acute and chronic tests for Brachydanio rerio at a concentration of 100 mg L(-1) , indicating a low toxicity of ACTs for this organism. The calculated LC50 (14 days, acute test) and NOEC (56 day chronic reproduction) for earthworms were >1000 and ≥250 mg kg(-1) soil respectively. CONCLUSIONS: The present study has provided baseline information needed to assess the potential hazard of the fungal toxins of A. caulina. In particular, the acute and chronic effects on aquatic (algae, Daphnia and fish) and terrestrial organisms (earthworms) have been highlighted. Finally, a comparison of the inherent toxicity of ACTs with other synthetic herbicides has shown comparative ecotoxicity of the tested mixture.

Rimonabant, a cannabinoid CB1 receptor antagonist, attenuates mechanical allodynia and counteracts oxidative stress and nerve growth factor deficit in diabetic mice.

Diabetes is one of the leading causes of painful neuropathy and to date, besides a tight glycemic control, a viable treatment for this complication is not available. Rimonabant is a selective cannabinoid CB(1) receptor antagonist that produces a significant increase in insulin sensitivity and a reduction of HbA(1c) in diabetic patients. This study aimed to investigate the therapeutic potential of rimonabant in relieving diabetes-induced neuropathic pain. The repeated treatment with rimonabant evoked a significant attenuation of mechanical allodynia in diabetic mice that was dose- and time-dependent. This effect occurred without alteration of hyperglycemia, but it was associated with significant effects on many key players in the pathogenesis of diabetic neuropathy. Metabolic changes induced by hyperglycemia lead to oxidative stress, deregulation of cytokine control and reduced production and transport of nerve growth factor (NGF), and all these factors contribute to neuropathic pain. Rimonabant treatment reduced oxidative stress in peripheral nerve, as revealed by the ability of the compound to counteract the reduced glutathione (GSH) depletion. The same repeated treatment inhibited tumor necrosis factor (TNFalpha) overproduction in the spinal cord and increased the NGF support. This rimonabant-induced improvement might favour the nerve regeneration; accordingly, the histological analysis of sciatic nerves showed a marked degeneration of myelinated fibers in diabetic mice, that was substantially reduced after rimonabant administration. These findings support the hypothesis that CB(1) antagonists would represent a new opportunity for diabetic patients, since currently there are no treatments for painful diabetic neuropathy other than treating the diabetic condition per se.

Soil genotoxicity assessment: a new stategy based on biomolecular tools and plant bioindicators.

The setting up of efficient early warning systems is a challenge to research for preventing environmental alteration and human disease. In this paper, we report the development and the field application of a new biomonitoring methodology for assessing soil genotoxicity. In the first part, the use of amplified fragment length polymorphism and flow cytometry techniques to detect DNA damage induced by soils artificially contaminated with heavy metals as potentially genotoxic compounds is explained. Results show that the combination of the two techniques leads to efficient detection of the sublethal genotoxic effect induced in the plant bioindicator by contaminated soil. By contrast, the classic mortality, root, and shoot growth vegetative endpoints prove inappropriate for assessing soil genotoxicity because, although they cause genotoxic damage, some heavy metals do not affect sentinel plant development negatively. The statistical elaboration of the data obtained led to the development of a statistical predictive model which differentiates four different levels of soil genotoxic pollution and can be used everywhere. The second part deals with the application of the biomonitoring protocol in the genotoxic assessment of two areas surrounding a steelworks in northern Italy and the effectiveness of this methodology. In this particular case, in these areas, the predictive model reveals a pollution level strictly correlated to the heavy metal concentrations revealed by traditional chemical analysis.