Bioremediation of HCH present in soil by the white-rot fungus Bjerkandera adusta in a slurry batch bioreactor

In the soil remediation process, the hydrophobic characteristics of pollutants and their affinity for soil matrix may be responsible for mass transfer limitations. The degradation of hexachlorocyclohexane (HCH) isomers present in a spiked soil by the white-rot Bjerkandera adusta was evaluated in a slurry system. Experiments in shaken flasks were performed to evaluate the action of the endogenous microflora, the adsorption of HCH on the fungal biomass and the potential synergic or antagonic actions between the microflora and the fungal biomass. The fungus significantly degraded the HCH isomers from the soil slurry in the following order: α≈γ>δ>β-HCH. The degradation process was further scaled in a 5-l reactor, where the solid load and concentration of the pollutant in the soil were evaluated. At optimal conditions, 100 g soil l⁻¹ and 100 mg total HCH l⁻¹, maximal degradations of 94.5%, 78.5% and 66.1% were attained after 30 d for γ-, α- and δ-HCH isomers, respectively, representing between 1.7 and 3.1-fold the values obtained at small scale. These results indicate that minimising mass transfer resistances is a key factor for HCH degradation from soil.     

Pesticide detection with a liposome-based nano-biosensor

Monitoring of the organophosphorus pesticides dichlorvos and paraoxon at very low levels has been achieved with liposome-based nano-biosensors. The enzyme acetylcholinesterase was effectively stabilized within the internal nano-environment of the liposomes. Within the liposomes, the pH sensitive fluorescent indicator pyranine was also immobilized for the optical transduction of the enzymatic activity. Increasing amounts of pesticides lead to the decrease of the enzymatic activity for the hydrolysis of the acetylcholine and thus to a decrease in the fluorescent signal of the pH indicator. The decrease of the liposome biosensors signal is relative to the concentration of dichlorvos and paraoxon down to 10⁻¹⁰ M levels. This biosensor system has been applied successfully to the detection of total toxicity in drinking water samples. Also a colorimetric screening device for pesticide analysis has been evaluated.     

Spatial and temporal variability of macroinvertebrate communities in two farmed Mediterranean rice fields

The spatial and temporal variation of macroinvertebrate assemblages was studied in two Portuguese commercial rice agroecosystems under the effect of field management involving the application of pesticides and fertilizers. A faunal succession of organisms was observed on both fields. Grazers were the first to colonize the paddies after a dry period when pesticides were applied, followed by development into nymphs and by an increase in the abundance of the species after the application of fertilizers. At the end of the season when no pesticides or fertilizers were applied, the communities changed with the presence of adult predators as a result of an increase in prey. Insecticide application revealed specific taxa increase due to the lack of competition with the target organism. Macroinvertebrates tended to prefer infested field margins with aquatic, submerged vegetation, revealing a spatial distribution along the paddies. Two different sampling devices were used and proved necessary in documenting the macroinvertebrate communities (grab for benthic and hand-net for pelagic organisms).     

Site-specific immobilization of a (His)6-tagged acetylcholinesterase on nickel nanoparticles for highly sensitive toxicity biosensors

This paper reports site-specific affinity immobilization of (His)6-tagged acetylcholinesterase (AChE) onto Ni/NiO nanoparticles for the development of an electrochemical screen-printed biosensor for the detection of organophosphate pesticides. The method is based on the specific affinity binding of the His-tagged enzyme to oxidized nickel nanoparticle surfaces in the absence of metal chelators. This approach allows stable and oriented attachment of the enzyme onto the oxidized nickel through the external His residue in one-step procedure, allowing for fast and sensitive detection of paraoxon in the concentration range from 10⁻⁸ to 10⁻¹³ M. A detection limit of 10⁻¹² M for paraoxon was obtained after 20 min incubation. This method can be used as a generic approach for the immobilization of other His-tagged enzymes for the development of biosensors.     

Simultaneous degradation of the pesticides methyl parathion and chlorpyrifos by an isolated bacterial consortium from a contaminated site

The simultaneous degradation of the pesticide methyl parathion and chlorpyrifos was tested using a bacterial consortium obtained by selective enrichment from highly contaminated soils in Moravia (Medellin, Colombia). Microorganisms identified in the consortium were Acinetobacter sp, Pseudomonas putida, Bacillus sp, Pseudomonas aeruginosa, Citrobacter freundii, Stenotrophomonas sp, Flavobacterium sp, Proteus vulgaris, Pseudomonas sp, Acinetobacter sp, Klebsiella sp and Proteus sp. In culture medium enriched with each of the pesticides, the consortium was able to degrade 150 mg l⁻¹ of methyl parathion and chlorpyrifos in 120 h. When a mixture of 150 mg l⁻¹ of both pesticides was used the percentage decreased to 72% for methyl parathion and 39% for chlorpyrifos. With the addition of glucose to the culture medium, the consortium simultaneously degraded 150 mg l⁻¹ of the pesticides in the mixture. 4 treatments were carried out in soil that included the addition of glucose with microorganisms, the addition of sugar cane with microorganisms, microorganisms without nutrient addition and without the addition of any item. In the treatment in which glucose was used, degradation percentages of methyl parathion and chlorpyrifos of 98% and 97% respectively were obtained in 120 h. This treatment also achieved the highest percentage of reduction in toxicity, monitored with Vibrio fischeri.     

Size selectivity of aqueous pores in stomatous cuticles of <Emphasis Type="Italic">Vicia faba</Emphasis> leaves

Size selectivity of aqueous pores in Vicia leaf cuticles was investigated by measuring the penetration of calcium salts into the abaxial surface of detached leaves. Molecular weights of salts ranged from 111 g mol^–1 to 755 g mol^–1. Penetration in light at 20°C and 100% humidity was a first order process and rate constants of penetration ranged from 0.39 h^–1 (CaCl₂) to 0.058 h^–1 (Ca-lactobionate). Penetration was a first order process in the dark as well, but the rate constants were smaller by a factor of 1.82. Plotting logarithmatised rate constants versus anhydrous molecular weights resulted in straight lines both in light and in the dark. The slopes per hour were very similar and the average slope was –1.2×10^–3 mol g^–1. Hence, size selectivity was not affected by stomatal opening, and in light or darkness permeability of Vicia cuticles decreased by a factor of 2.9 when molecular weight increased from 100 g mol^–1 to 500 g mol^–1. Silver nitrate was preferentially precipitated as silver chloride in guard cells, glandular trichomes and at the base of trichomes. It was concluded that these precipitates mark the location of aqueous pores in Vicia leaf cuticles. The size selectivity of aqueous pores in Vicia leaf cuticles is small compared to that observed in poplar leaf cuticles, in which permeability decreased by a factor of 7–13 for the same range of molecular weights. It is also much smaller than size selectivity of the lipophilic pathway in cuticles. These findings suggest that active ingredients of pesticides, growth regulators and chemical inducers with high molecular weights penetrate leaves at higher rates when formulated as ions.     

Molecular characterization and expression of the gene encoding aspartate aminotransferase from the Pacific oyster Crassostrea gigas exposed to environmental stressors

A partial cDNA encoding cytosolic aspartate aminotransferase (AST) (EC 2.6.1.1) was isolated from a Crassostrea gigas digestive gland library. This sequence was used to design specific primers to amplify the AST genomic sequence. We obtained a complete gene, 5054 bp in length, encoding cytosolic AST and containing a 404 amino acid open reading frame. Phylogenetic analysis showed that C. gigas AST sequence constitutes a branch distinct from homologous sequences from other invertebrate groups. We also investigated AST mRNA expression in different tissues of oysters exposed to hydrocarbons, pesticides, hypoxia and hypo-salinity stress. The results showed that AST expression responds to hydrocarbon exposure, hypoxia and salinity stress, but not to pesticide exposure in an organ and time-specific manner. Use of AST as a potential molecular biomarker for monitoring of disturbed ecosystems is discussed.     

Chromatographic evaluation of the toxicity in fish of pesticides

Ecotoxicity assessment is essential before placing new chemical substances on the market. An investigation of the use of the chromatographic retention (log k) in biopartitioning micellar chromatography (BMC) as an in vitro approach to evaluate the toxicity in fish of pesticides (acute toxicity levels as pLC(50)) is proposed. A heterogeneous data set of 85 pesticides from six chemical families with available experimental fish toxicity data (ECOTOX database from U.S. Environmental Protection Agency (EPA)) was used. For pesticides exhibiting non-polar narcosis mechanism in fish (non-specific toxicity), more reliable models and precise pLC(50) estimations are obtained from log k (quantitative retention-activity relationships, QRAR) than from log P (quantitative structure-activity relationships, QSAR) or ECOSAR (ECOSAR program from U.S. EPA).     

表面残留杀虫剂对蜘蛛的影响研究

以前的报道多集中于研究杀虫剂直接喷洒对蜘蛛的影响。本研究以三突花蛛和沟渠豹蛛为对象,测定了溴氰菊酯、毒死蜱和查虫清（植物源杀虫剂）共3种杀虫剂的表面残留对蜘蛛的驱避作用和急性毒性。结果表明：残留2h的毒死蜱和查虫清对三突花蛛无驱避作用,而残留2h的溴氰菊酯对三突花蛛有明显地驱避作用。3种杀虫剂的2h表面残留对沟渠豹蛛均有显著的驱避作用,其中溴氰菊酯的驱避作用最强。残留24h的3种杀虫剂对三突花蛛和沟渠豹蛛无明显的驱避作用。表面残留的毒死蜱和查虫清对蜘蛛的毒性小,溴氰菊酯的残留对三突花蛛和沟渠豹蛛毒性大。表面残留2h的溴氰菊酯对沟渠豹蛛的毒性显著高于残留24h的溴氰菊酯。     

Carboxylesterase activities toward pesticide esters in crops and weeds

Proteins were extracted from maize, rice, sorghum, soybean, flax and lucerne; the weeds Abutilon theophrasti, Echinochloa crus-galli, Phalaris canariensis, Setaria faberii, Setaria viridis, Sorghum halepense and the model plant Arabidopsis thaliana and assayed for carboxylesterase activity toward a range of xenobiotics. These included the pro-herbicidal esters clodinafop-propargyl, fenoxaprop-ethyl, fenthioprop-ethyl, methyl-2,4-dichlorophenoxyacetic acid (2,4-d-methyl), bromoxynil-octanoate, the herbicide-safener cloquintocet-mexyl and the pyrethroid insecticide permethrin. Highest activities were recorded with alpha-naphthyl acetate and methylumbelliferyl acetate. Esters of p-nitrophenol were also readily hydrolysed, with turnover declining as the chain length of the acyl component increased. Activities determined with model substrates were much higher than those observed with pesticide esters and were of limited value in predicting the relative rates of hydrolysis of the crop protection agents. Substrate preferences with the herbicides were typically 2,4-d-methyl>clodinafop-propargyl>fenthioprop-ethyl, fenoxaprop-ethyl and bromoxynil-octanoate. Isoelectric focussing in conjunction with staining for esterase activity using alpha-naphthyl acetate as substrate confirmed the presence of multiple carboxylesterase isoenzymes in each plant, with major qualitative differences observed between species. The presence of serine hydrolases among the resolved isoenzymes was confirmed through their selective inhibition by the organophosphate insecticide paraoxon. Our studies identify potentially exploitable differences between crops and weeds in their ability to bioactivate herbicides by enzymic hydrolysis and also highlight the usefulness of Arabidopsis as a plant model to study xenobiotic biotransformation.