Phenol tolerance, changes of antioxidative enzymes and cellular damage in transgenic tobacco hairy roots colonized by arbuscular mycorrhizal fungi

Phytoremediation has been recognized as a cheap and eco-friendly technology which could be used for the remediation of organic pollutants, such as phenolic compounds. Besides, the extent to which plants react to environmental pollution might depend on rhizosphere processes such as mycorrhizal symbiosis. In the present work, phenol tolerance of transgenic tobacco hairy roots (HR), namely TPX1, colonized with an arbuscular mycorrhizal fungus (AMF) was studied. However, the question is whether AMF symbiosis can moderate adverse effects of phenol to the plant tissues. Thus, the antioxidative response as well as parameters of oxidative damage, like malondialdehyde (MDA) content, were determined. Antioxidative enzymes such as peroxidase, superoxide dismutase, ascorbate peroxidase were higher in TPX1 HR colonized with AMF, compared to wild type HR colonized by AMF, in the presence of increasing concentrations of the pollutant. Besides, MDA levels remained unaltered in TPX1 HR associated with AMF treated with the xenobiotic. These results, suggested that this culture could tolerate phenol and moreover, it has an efficient protective mechanism against phenol-induced oxidative damage, which is of great importance in the selection of species with remediation capacities. Thus, transgenic HR colonized with AMF could be considered as an interesting model system to study different processes which play a key role in the phytoremediation of organic pollutants.     

The microbiota of an unpolluted calcareous soil faces up chlorophenols: Evidences of resistant strains with potential for bioremediation

To highlight the effects of a variety of chlorophenols (CP) in relation to the response of an indigenous bacterial community, an agricultural Mediterranean calcareous soil has been studied in microcosms incubated under controlled laboratory conditions. Soil samples were artificially polluted with 2-monochlorophenol (MCP), 2,4,6-trichlorophenol (TCP) and pentachlorophenol (PCP), at concentrations ranging from 0.1 up to 5000 mg kg⁻¹. Both activity and composition of the microbial community were assessed during several weeks, respectively, by respirometric methods and PCR-DGGE analysis of extracted DNA and RNA. Significant decreases in soil respirometric values and changes in the bacterial community composition were observed at concentrations above 1000 mg kg⁻¹ MCP and TCP, and above 100 mg kg⁻¹ PCP. However, the persistence of several active bacterial populations in soil microcosms contaminated with high concentration of CP, as indicated by DGGE fingerprints, suggested the capacity of these native bacteria to survive in the presence of the pollutants, even without a previous adaptation or contact with them.The isolation of potential CP degraders was attempted by culture plating from microcosms incubated with high CP concentrations. Twenty-three different isolates were screened for their resistance to TCP and PCP. The most resistant isolates were identified as Kocuria palustris, Lysobacter gummosus, Bacillus sp. and Pseudomonas putida, according to 16S rRNA gene homology. In addition, these four isolates also showed the capacity to reduce the concentration of TCP and PCP from 15% to 30% after 5 d of incubation in laboratory assays (initial pollutant concentration of 50 mg L⁻¹). Isolate ITP29, which could be a novel species of Bacillus, has been revealed as the first known member in this bacterial group with potential for CP bioremediation applications, usually wide-spread in the soil natural communities, which has not been reported to date as a CP degrader.     

Toxicity of 58 substituted anilines and phenols to algae Pseudokirchneriella subcapitata and bacteria Vibrio fischeri: comparison with published data and QSARs

A congeneric set of 58 substituted anilines and phenols was tested using the 72-h algal growth inhibition assay with Pseudokirchneriella subcapitata and 15-min Vibrio fischeri luminescence inhibition assay. The set contained molecules substituted with one, two or three groups chosen from -chloro, -methyl or -ethyl. For 48 compounds there was no REACH-compatible algal toxicity data available before. The experimentally obtained EC50 values (mg L⁻¹) for algae ranged from 1.43 (3,4,5-trichloroaniline) to 197 (phenol) and for V. fischeri from 0.37 (2,3,5-trichlorophenol) to 491 (aniline). Only five of the tested 58 chemicals showed inhibitory effect to algae at concentrations >100 mg L⁻¹, i.e. could be classified as “not harmful”, 32 chemicals as “harmful” (10-100 mg L⁻¹) and 21 as “toxic” (1-10 mg L⁻¹). The occupied para-position tended to increase toxicity whereas most of the ortho-substituted congeners were the least toxic. As a rule, the higher the number of substituents the higher the hydrophobicity and toxicity. However, in case of both assays, the compounds of similar hydrophobicity showed up to 30-fold different toxicities. There were also assay/organism dependent tendencies: phenols were more toxic than anilines in the V. fischeri assay but not in the algal test. The comparison of the experimental toxicity data to the data available from the literature as well as to QSAR predictions showed that toxicity of phenols to algae can be modeled based on hydrophobicity, whereas the toxicity of anilines to algae as well as toxicity of both anilines and phenols to V. fischeri depended on other characteristics in addition to logK_ow.     

Studies on atmospheric degradation of diazinon in the EUPHORE simulation chamber

The gas phase atmospheric degradation of diazinon has been investigated at the large outdoor European Photoreactor (EUPHORE) in Valencia, Spain. The rate constant for reaction of diazinon with OH radicals was measured using a conventional relative rate method with di-n-buthylether as reference compound being k = (3.5 ± 1.2) × 10⁻¹¹ cm³ molecule⁻¹ s⁻¹ at 302 ± 10 K and atmospheric pressure. The available evidence indicates that tropospheric degradation of diazinon is mainly controlled by reaction with OH radicals, and that the tropospheric lifetime with respect to the OH reaction is estimated to be around 4 h whereas its lifetime with respect to the photolysis is higher than 1 d under our conditions. Significant aerosol formation was observed following the reaction of diazinon with OH radicals, and the main carbon-containing products detected in the particle phase were hydroxydiazinon, hydroxydiazoxon and 2-isopropyl-6-methyl-pyrimidinyl-4-ol.     

Photolytic and photocatalytic degradation of 6-chloronicotinic acid

This work describes for the first time the photolytic and photocatalytic degradation of 6-chloronicotinic acid (6CNA) in double deionised water, which is a degradation product of neonicotinoid insecticides imidacloprid and acetamiprid, and it is known to appear in different environmental matrices. Photolytic experiments were performed with three UVA (ultraviolet A) polychromatic fluorescent lamps with broad maximum at 355 nm, while photocatalytic experiments were performed using immobilised titanium dioxide (TiO₂) on six glass slides in the spinning basket inside a photocatalytic quartz cell under similar irradiation conditions. Photolytic degradation revealed no change in concentration of 6CNA within 120 min of irradiation, while the photocatalytic degradation within 120 min, obeyed first-order kinetics. The observed disappearance rate constant was k = 0.011 ± 0.001 min⁻¹ and t_1/2 was 63.1 ± 5.5 min. Mineralisation rate was estimated through total organic carbon (TOC) and measurements revealed no carbon removal in case of photolysis after 120 min of exposure. However in photocatalytic experiments 46 ± 7% mineralisation was achieved within 120 min of irradiation. Nevertheless, the removal of total nitrogen (TN) was not observed across all experiments. Ion chromatographic analyses indicated transformation of chlorine atoms to chloride and increase of nitrate(V) ions only via photocatalytic experiments. Efficiency of selected advanced oxidation process (AOP) was investigated through toxicity assessment with Vibrio fischeri luminescent bacteria and revealed higher adverse effects of treated samples on bacteria following photocatalytic degradation in spite of the fact that higher mineralisation was achieved. New hydroxylated product generated in photocatalytic experiments with TiO₂, was confirmed with liquid chromatography-electro spray ionisation mass spectrometry (LC-ESI-MS/MS) analyses, gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy (¹H NMR).     

Geographic and trophic patterns of OCs in pelagic seabirds from the NE Atlantic and the Mediterranean: a multi-species/multi-locality approach

Trophic ecology and geographic location are crucial factors explaining OC levels in marine vertebrates, but these factors are often difficult to disentangle. To examine their relative influence, we analyzed PCBs, DDTs and stable-nitrogen isotope signatures (δ15N) in the blood of 10 pelagic seabird species across 7 breeding localities from the northeast Atlantic and western Mediterranean. Large scale geographic patterns emerged due to the confined character and greater historical OC inputs in the Mediterranean compared to the Atlantic basin. Spatial patterns also emerged at the regional scale within the Atlantic basin, probably associated with long-range pollutant transport. Trophic ecology, however, was also a major factor explaining OC levels. We found clear and consistent OC differences among species regardless of the sampled locality. However, species δ15N and blood OC levels were not correlated within most breeding localities. Petrel species showed significantly greater OC burdens than most shearwater species but similar trophic positions, as indicated by their similar δ15N signatures. This pattern probably results from Petrel species feeding on mesopelagic fish and squid that migrate close to the sea surface at night, whereas shearwater species mainly feed on epipelagic diurnal prey. In sum, this study illustrates the lasting and unequal influence of past human activities such as PCB and DDT usage across different marine regions. In addition, our results suggest that multi-species designs are powerful tools to monitor geographic patterns of OCs and potentially useful to assess their vertical dynamics in the marine environment.     

Ozone flux over a Norway spruce forest and correlation with net ecosystem production

Daily ozone deposition flux to a Norway spruce forest in Czech Republic was measured using the gradient method in July and August 2008. Results were in good agreement with a deposition flux model. The mean daily stomatal uptake of ozone was around 47% of total deposition. Average deposition velocity was 0.39 cm s⁻¹ and 0.36 cm s⁻¹ by the gradient method and the deposition model, respectively. Measured and modelled non-stomatal uptake was around 0.2 cm s⁻¹. In addition, net ecosystem production (NEP) was measured by using Eddy Covariance and correlations with O₃ concentrations at 15 m a.g.l., total deposition and stomatal uptake were tested. Total deposition and stomatal uptake of ozone significantly decreased NEP, especially by high intensities of solar radiation.     

Intercomparison exercise for gases emitted by a cement industry in Spain: a functional data approach

This paper describes the results of an intercomparison exercise referring to the measurement of atmospheric pollutants emitted by cement plants. The research was conducted in 2008 in Catalonia, Spain. Thirteen Spanish companies accredited to make pollutant measurements and with suitably approved equipment and trained staff participated in the research. The aim of the research was to evaluate the technical competence of the accredited companies in conducting tests in situ. The results obtained from the application of the methodology described in international standards--on the basis of conventional statistical analyses--were compared with those obtained using functional data analysis. On comparing the two methods, it was concluded that the functional method, although more complex, has advantages over the conventional statistical method. This is because the functional approach compares curves rather than mean values. Furthermore, it does not presuppose a normal distribution of the data.     

Soil bioassays as tools for sludge compost quality assessment

Composting is a waste management technology that is becoming more widespread as a response to the increasing production of sewage sludge and the pressure for its reuse in soil. In this study, different bioassays (plant germination, earthworm survival, biomass and reproduction, and collembolan survival and reproduction) were assessed for their usefulness in the compost quality assessment.Compost samples, from two different composting plants, were taken along the composting process, which were characterized and submitted to bioassays (plant germination and collembolan and earthworm performance). Results from our study indicate that the noxious effects of some of the compost samples observed in bioassays are related to the low organic matter stability of composts and the enhanced release of decomposition endproducts, with the exception of earthworms, which are favored. Plant germination and collembolan reproduction inhibition was generally associated with uncomposted sludge, while earthworm total biomass and reproduction were enhanced by these materials. On the other hand, earthworm and collembolan survival were unaffected by the degree of composting of the wastes. However, this pattern was clear in one of the composting procedures assessed, but less in the other, where the release of decomposition endproducts was lower due to its higher stability, indicating the sensitivity and usefulness of bioassays for the quality assessment of composts.     

Determinants of efficiency in the provision of municipal street-cleaning and refuse collection services

Effective waste management systems can make critical contributions to public health, environmental sustainability and economic development. The challenge affects every person and institution in society, and measures cannot be undertaken without data collection and a quantitative analysis approach. In this paper, the two-stage double bootstrap procedure of Simar and Wilson (2007) is used to estimate the efficiency determinants of Spanish local entities in the provision of public street-cleaning and refuse collection services. The purpose is to identify factors that influence efficiency. The final sample comprised 1072 municipalities. In the first stage, robust efficiency estimates are obtained with Data Envelopment Analysis (DEA). We apply the second stage, based on a truncated-regression, to estimate the effect of a group of environmental factors on DEA estimates. The results show the existence of a significant relation between efficiency and all the variables analysed (per capita income, urban population density, the comparative index of the importance of tourism and that of the whole economic activity). We have also considered the influence of a dummy categorical variable - the political sign of the governing party - on the efficient provision of the services under study. The results from the methodology proposed show that municipalities governed by progressive parties are more efficient.