Impact of metal stress on the production of secondary metabolites in <Emphasis Type="Italic">Pteris vittata</Emphasis> L. and associated rhizosphere bacterial communities

Plants adapt to metal stress by modifying their metabolism including the production of secondary metabolites in plant tissues. Such changes may impact the diversity and functions of plant associated microbial communities. Our study aimed to evaluate the influence of metals on the secondary metabolism of plants and the indirect impact on rhizosphere bacterial communities. We then compared the secondary metabolites of the hyperaccumulator Pteris vittata L. collected from a contaminated mining site to a non-contaminated site in Vietnam and identified the discriminant metabolites. Our data showed a significant increase in chlorogenic acid derivatives and A-type procyanidin in plant roots at the contaminated site. We hypothesized that the intensive production of these compounds could be part of the antioxidant defense mechanism in response to metals. In parallel, the structure and diversity of bulk soil and rhizosphere communities was studied using high-throughput sequencing. The results showed strong differences in bacterial composition, characterized by the dominance of Proteobacteria and Nitrospira in the contaminated bulk soil, and the enrichment of some potential human pathogens, i.e., Acinetobacter, Mycobacterium, and Cupriavidus in P. vittata’s rhizosphere at the mining site. Overall, metal pollution modified the production of P. vittata secondary metabolites and altered the diversity and structure of bacterial communities. Further investigations are needed to understand whether the plant recruits specific bacteria to adapt to metal stress.     

Cadmium-tolerant bacteria induce metal stress tolerance in cereals

Cadmium usually hampers plant growth, but bacterial inoculation may improve stress tolerance in plants to Cd by involving various mechanisms. The objective was to characterize and identify bacteria that improve plant growth under Cd stress and reduce Cd uptake. Cadmium-tolerant bacteria were isolated from rhizosphere soil, which was irrigated with tannery effluent, and six strains were selected as highly tolerant to Cd, showing minimum inhibitory concentration as 500 mg L^?1 or 4.45 mmol L^?1. These strains were identified by 16S rRNA gene analysis and functional analysis in regard to plant growth promotion characteristics. To determine their effect on cereal growth under Cd stress, seeds were inoculated with these strains individually and grown in soil contaminated with three Cd levels (0, 40 and 80 mg kg^?1). Biomass production, relative water content (RWC), electrolyte leakage (ELL) and tissue Cd concentration were measured. Biomass of both cereals was inhibited strongly when exposed to Cd; however, bacterial inoculation significantly reduced the suppressive effect of Cd on cereal growth and physiology. The bacterial isolates belonged to the genera Klebsiella, Stenotrophomonas, Bacillus and Serratia. Maize was more sensitive than wheat to Cd. Klebsiella sp. strain CIK-502 had the most pronounced effects in promoting maize and wheat growth and lowering Cd uptake under Cd stress.     

Response of bacterial pdo1, nah,and C12O genes to aged soil PAH pollution in a coke factory area

Soil pollution caused by polycyclic aromatic hydrocarbons (PAHs) is threatening human health and environmental safety. Investigating the relative prevalence of different PAH-degrading genes in PAH-polluted soils and searching for potential bioindicators reflecting the impact of PAH pollution on microbial communities are useful for microbial monitoring, risk evaluation, and potential bioremediation of soils polluted by PAHs. In this study, three functional genes, pdo1, nah, and C12O, which might be involved in the degradation of PAHs from a coke factory, were investigated by real-time quantitative PCR (qPCR) and clone library approaches. The results showed that the pdo1 and C12O genes were more abundant than the nah gene in the soils. There was a significantly positive relationship between the nah or pdo1 gene abundances and PAH content, while there was no correlation between C12O gene abundance and PAH content. Analyses of clone libraries showed that all the pdo1 sequences were grouped into Mycobacterium, while all the nah sequences were classified into three groups: Pseudomonas, Comamonas, and Polaromonas. These results indicated that the abundances of nah and pdo1 genes were positively influenced by levels of PAHs in soil and could be potential microbial indicators reflecting the impact of soil PAH pollution and that Mycobacteria were one of the most prevalent PAHs degraders in these PAH-polluted soils. Principal component analysis (PCA) and correlation analyses between microbial parameters and environmental factors revealed that total carbon (TC), total nitrogen (TN), and dissolved organic carbon (DOC) had positive effects on the abundances of all PAH-degrading genes. It suggests that increasing TC, TN, and DOC inputs could be a useful way to remediate PAH-polluted soils.     

Assessing the hydrocarbon degrading potential of indigenous bacteria isolated from crude oil tank bottom sludge and hydrocarbon-contaminated soil of Azzawiya oil refinery,Libya

The disposal of hazardous crude oil tank bottom sludge (COTBS) represents a significant waste management burden for South Mediterranean countries. Currently, the application of biological systems (bioremediation) for the treatment of COTBS is not widely practiced in these countries. Therefore, this study aims to develop the potential for bioremediation in this region through assessment of the abilities of indigenous hydrocarbonoclastic microorganisms from Libyan Hamada COTBS for the biotreatment of Libyan COTBS-contaminated environments. Bacteria were isolated from COTBS, COTBS-contaminated soil, treated COTBS-contaminated soil, and uncontaminated soil using Bushnell Hass medium amended with Hamada crude oil (1 %) as the main carbon source. Overall, 49 bacterial phenotypes were detected, and their individual abilities to degrade Hamada crude and selected COBTS fractions (naphthalene, phenanthrene, eicosane, octadecane and hexane) were evaluated using MT2 Biolog plates. Analyses using average well colour development showed that ~90 % of bacterial isolates were capable of utilizing representative aromatic fractions compared to 51 % utilization of representative aliphatics. Interestingly, more hydrocarbonoclastic isolates were obtained from treated contaminated soils (42.9 %) than from COTBS (26.5 %) or COTBS-contaminated (30.6 %) and control (0 %) soils. Hierarchical cluster analysis (HCA) separated the isolates into two clusters with microorganisms in cluster 2 being 1.7- to 5-fold better at hydrocarbon degradation than those in cluster 1. Cluster 2 isolates belonged to the putative hydrocarbon-degrading genera; Pseudomonas, Bacillus, Arthrobacter and Brevundimonas with 57 % of these isolates being obtained from treated COTBS-contaminated soil. Overall, this study demonstrates that the potential for PAH degradation exists for the bioremediation of Hamada COTBS-contaminated environments in Libya. This represents the first report on the isolation of hydrocarbonoclastic bacteria from Libyan COTBS and COTBS-contaminated soil.     

Standard inocula preparations reduce the bacterial diversity and reliability of regulatory biodegradation tests

OECD ready biodegradability tests have been central to understanding the biodegradation of chemicals from a regulatory perspective for many decades. They are not fit for contemporary prioritisation of chemicals based on persistence, however, due to the low concentration of inocula used, short duration and high variability between tests. Two OECD standard inoculum pretreatment methods (settlement and filtration) were investigated to observe their effect on the probability of biodegradation and associated changes in bacterial community structure and diversity of inocula sourced from the activated sludge process of wastewater treatment plants. Both settlement and filtration were shown to dramatically and significantly reduce the probability and increase the variability of biodegradation of 4-nitrophenol compared to the use of unprocessed inocula. These differences were associated with a significant hundred-fold reduction in cell numbers and solids content and a significant shift in bacterial community structure that was sometimes accompanied by significant reductions in detectable operational taxonomic unit richness and evenness. The natural variation (between different environments) and variation due to differential selection of bacterial communities (by different pretreatment methods) is offered as an explanation for the historical high variability in standard OECD ready biodegradability tests.     

EDTA ameliorates phytoextraction of lead and plant growth by reducing morphological and biochemical injuries in Brassica napus L. under lead stress

Brassica species are very effective in remediation of heavy metal contaminated sites. Lead (Pb) as a toxic pollutant causes number of morphological and biochemical variations in the plants. Synthetic chelator such as ethylenediaminetetraacetic acid (EDTA) improves the capability of plants to uptake heavy metals from polluted soil. In this regard, the role of EDTA in phytoextraction of lead, the seedlings of Brassica napus L. were grown hydroponically. Lead levels (50 and 100 μM) were supplied alone or together with 2.5 mM EDTA in the nutrient culture. After 7 weeks of stress, plants indicated that toxicity of Pb caused negative effects on plants and significantly reduced growth, biomass, chlorophyll content, gas exchange characteristics, and antioxidant enzymes activities such as superoxide dismutase (SOD), guaiacol peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT). Exposure to Pb induced the malondialdehyde (MDA), and hydrogen peroxide (H₂O₂) generation in both shoots and roots. The addition of EDTA alone or in combination with Pb significantly improved the plant growth, biomass, gas exchange characteristics, chlorophyll content, and antioxidant enzymes activities. EDTA also caused substantial improvement in Pb accumulation in Brassica plants. It can be deduced that application of EDTA significantly lessened the adverse effects of lead toxicity. Additionally, B. napus L. exhibited greater degree of tolerance against Pb toxicity and it also accumulated significant concentration of Pb from media.     

Toxicity assessment through multiple endpoint bioassays in soils posing environmental risk according to regulatory screening values

Toxicity profiles of two soils (a brownfield in Legazpi and an abandoned iron mine in Zugaztieta; Basque Country) contaminated with several metals (As, Zn, Pb and Cu in Legazpi; Zn, Pb, Cd and Cu in Zugaztieta) and petroleum hydrocarbons (in Legazpi) were determined using a multi-endpoint bioassay approach. Investigated soils exceeded screening values (SVs) of regulatory policies in force (Basque Country; Europe). Acute and chronic toxicity bioassays were conducted with a selected set of test species (Vibrio fischeri, Dictyostelium discoideum, Lactuca sativa, Raphanus sativus and Eisenia fetida) in combination with chemical analysis of soils and elutriates, as well as with bioaccumulation studies in earthworms. The sensitivity of the test species and the toxicity endpoints varied depending on the soil. It was concluded that whilst Zugaztieta soil showed very little or no toxicity, Legazpi soil was toxic according to almost all the toxicity tests (solid phase Microtox®, D. discoideum inhibition of fruiting body formation and developmental cycle solid phase assays, lettuce seed germination and root elongation test, earthworm acute toxicity and reproduction tests, D. discoideum cell viability and replication elutriate assays). Thus, albeit both soils had similar SVs, their ecotoxicological risk, and therefore the need for intervening, was different for each soil as unveiled after toxicity profiling based on multiple endpoint bioassays. Such a toxicity profiling approach is suitable to be applied for scenario-targeted soil risk assessment in those cases where applicable national/regional soil legislation based on SVs demands further toxicity assessment.     

Combined effects of DOM and biosurfactant enhanced biodegradation of polycylic armotic hydrocarbons (PAHs) in soil–water systems

This study systematically investigated the interactive effects of dissolved organic matter (DOM) and biosurfactant (rhamnolipid) on the biodegradation of phenanthrene (PHE) and pyrene (PYR) in soil–water systems. The degradations of two polycyclic aromatic hydrocarbons (PAHs) were fitted well with first order kinetic model and the degradation rates were in proportion to the concentration of biosurfactant. In addition, the degradation enhancement of PHE was higher than that of PYR. The addition of soil DOM itself at an environmental level would inhibit the biodegradation of PAHs. However, in the system with co-existence of DOM and biosurfactant, the degradation of PAHs was higher than that in only biosurfactant addition system, which may be attributed to the formation of DOM–biosurfactant complex micelles. Furthermore, under the combined conditions, the degradation of PAH increased with the biosurfactant concentration, and the soil DOM added system showed slightly higher degradation than the compost DOM added system, indicating that the chemical structure and composition of DOM would also affect the bioavailability of PAHs. The study result may broaden knowledge of biosurfactant enhanced bioremediation of PAHs contaminated soil and groundwater.     

Degradation of PAHs in soil by Lasiodiplodia theobromae and enhanced benzo[a]pyrene degradation by the addition of Tween-80

Benzo[a]pyrene (BaP), a five-ring polycyclic aromatic hydrocarbon (PAH), which has carcinogenic potency, is highly recalcitrant and resistant to microbial degradation. A novel fungus, Lasiodiplodia theobromae (L. theobromae), which can degrade BaP as a sole carbon source in liquid, was isolated in our laboratory. To prompt the further application of L. theobromae in remediation of sites polluted by BaP and other PAHs, the present study was targeted toward the removal of BaP and PAHs from soil by L. theobromae. The degradation of BaP by L. theobromae was studied using a soil spiked with 50 mg/kg BaP. L. theobromae could remove 32.1 % of the BaP after 35 days of cultivation. Phenanthrene (PHE) inhibited BaP degradation as a competitive substrate. The tested surfactants enhanced BaP degradation in soil by different extents, and a removal rate of 92.1 % was achieved at a Tween-80 (TW-80) concentration of 5 g/kg. It was revealed that TW-80 could not only enhance BaP bioavailability by increasing its aqueous solubility and decreasing the size of its colloid particles but also increase enzyme secretion from L. theobromae and the population of L. theobromae. Moreover, ergosterol content together with the biomass C indicated the increase in L. theobromae biomass during the BaP biodegradation process in soils. Finally, a soil from a historically PAH-contaminated field at Beijing Coking Plant in China was tested to assess the feasibility of applying L. theobromae in the remediation of polluted sites. The total removal rate of PAHs by L. theobromae was 53.3 %, which is 13.1 % higher than that by Phanerochaete chrysosporium (P. chrysosporium), an effective PAH degrader. The addition of TW-80 to the field soil further enhanced PAH degradation to 73.2 %. Hence, L. theobromae is a promising novel strain to be implemented in the remediation of soil polluted by PAHs.     

Mobility and phytoavailability of Cu,Cr, Zn,and As in a contaminated soil at a wood preservation site after 4 years of aided phytostabilization

The remediation of copper-contaminated soils by aided phytostabilisation in 16 field plots at a wood preservation site was investigated. The mobility and bioavailability of four potentially toxic trace elements (PTTE), i.e., Cu, Zn, Cr, and As, were investigated in these soils 4 years after the incorporation of compost (OM, 5 % w/w) and dolomite limestone (DL, 0.2 % w/w), singly and in combination (OMDL), and the transplantation of mycorrhizal poplar and willows. Topsoil samples were collected in all field plots and potted in the laboratory. Total PTTE concentrations were determined in soil pore water (SPW) collected by Rhizon soil moisture samplers. Soil exposure intensity was assessed by Chelex100-DGT (diffusive gradient in thin films) probes. The PTTE phytoavailability was characterized by growing dwarf beans on potted soils and analyzing their foliar PTTE concentrations. OM and DL, singly and in combination (OMDL), were effective to decrease foliar Cu, Cr, Zn, and As concentrations of beans, the lowest values being numerically for the OM plants. The soil treatments did not reduce the Cu and Zn mineral masses of the bean primary leaves, but those of Cr and As decreased for the OM and DL plants. The Cu concentration in SPW was increased in the OM soil and remained unchanged in the DL and OMDL soils. The available Cu measured by DGT used to assess the soil exposure intensity correlated with the foliar Cu concentration. The Zn concentrations in SPW were reduced in the DL soil. All amendments increased As in the SPW. Based on DGT data, Cu availability was reduced in both OM and OMDL soils, while DL was the most effective to decrease soil Zn availability.     

Occurrence of ectoparasitic arthropods associated with rodents in Hail region northern Saudi Arabia

Ectoparasitic arthropods are a diverse element of the Saudi fauna. Due to this, a survey of ectoparasites associated with rodents was conducted as a preliminary study in five districts of Hail region of northern Saudi Arabia for the first time. Ectoparasites extracted from 750 rodents were sampled and identified by recording their frequency of appearance. Results revealed that 1,287 ectoparasites infested 316 of the captured rodent hosts. These ectoparasites parasitized on four species of rodents including three species of rats Rattus rattus rattus, Rattus rattus frugivorus, and Rattus rattus alexandrinus and one species of mouse Acomys dimidiatus (Rodentia: Muridae). The ectoparasites belong to four different groups: ticks, fleas, lice, and mites. Ticks were the highest in the number, while fleas were the lowest among all the extracted ectoparasite groups. The collected ectoparasitic arthropods consisted of seven species. Ticks were of two species: Rhipicephalus turanicus and Rhipicephalus sanguineus (Acari: Ixodidae), fleas were of two species: Xenopsylla cheopis and Xenopsyllus conformis mycerini (Siphonaptera: Pulicidae), lice was a single species: Polyplax serrata (Anoplura: Hoplopleuridae), and mites were of two species: Laelaps nuttali and Laelaps echidninus (Mesostigmata: Laelapidae). The findings of the study showed that the intensity of infestation was varied between rodent host sexes, wherein females had the highest rate of parasitic infestation, and the parasitic index of appearance was very high for one group of parasites (i.e., ticks). The parasitic prevalence was 42.13 % on rodents, and mites were the most prevalent parasite species. Overall, this study was carried out to establish baseline data for ectoparasite-infested rodents in Hail region, Saudi Arabia, and may help for appropriate planning to control zoonotic diseases in this area.     

Semivolatile PAH and n-alkane gas/particle partitioning using the dual model: up-to-date coefficients and comparison with experimental data

The gas/particle partitioning coefficient K _p , of a semivolatile compound is a key parameter for its atmospheric fate. The most complete method of predicting K _p for polycyclic aromatic hydrocarbons (PAHs) is offered by the dual model, as it describes both the adsorption on soot and absorption into organic matter processes. However, experimental and model data exist almost exclusively for PAHs. In order to bridge this gap, experimental data on the phase partitioning of both PAHs and n-alkanes were collected at an urban and a remote site. Moreover, all the necessary parameters (e.g., octanol–air and soot–air partitioning coefficients) for the dual model have been collected and updated or (if missing) estimated for the first time. The results point out that both absorption and adsorption seem to contribute to the partitioning of PAHs and n-alkanes. However, it seems that the dual model always underestimates the particle sorption not only for PAHs but also for n-alkanes.     

Selection of suitable lichen bioindicator species for monitoring climatic variability in the Himalaya

Interspecific comparison in metals and PAHs profile in three lichen species, Flavoparmelia caperata, Phaeophyscia hispidula and Pyxine sorediata, were studied in different altitudinal gradients of the Western Himalayas. The species collected from 14 sites, enroute from Dehradun to Morinda (243 Km) including the trekking route 42 Km from Taluka to Morinda having an altitudinal gradient between 850–3,750 m, were analysed for their metals and PAHs. The species showed similar metal as well as PAHs profile under similar altitudinal gradients in the sequence of F. caperata > P. hispidula > P. sorediata. The difference in pollutant concentrations within each lichen species may be related to intrinsic attributes of the species, such as thallus morphology and the presence of lichen substances which are responsible for the sensitivity and accumulation potential of a particular species. Novelty of the present study lies on the fact that all the species show a similar efficiency of reflecting the environmental condition of the area, albeit the coefficient values of individual species for individual pollutant obtained by three-factor ANOVA revealed that the bioaccumulation affinity of F. caperata is significantly higher than P. hispidula and P. sorediata. For individual metals, F. caperata has a higher affinity for Al, Cr, Fe, Pb and Zn while P. hispidula has a significant positive affinity for Fe and Pb. PCA analysis of sites with respect to pollutant revealed the segregation of sites based on source and distance. Combining the bioaccumulation potential parameters along with geostatistical (GIS) techniques establishes that F. caperata species is a better accumulator of metals and PAHs in comparison to P. hispidula and P. sorediata in the temperate regions of the Himalaya.     

Environmental effects of soil contamination by shale fuel oils

Estonia is currently one of the leading producers of shale oils in the world. Increased production, transportation and use of shale oils entail risks of environmental contamination. This paper studies the behaviour of two shale fuel oils (SFOs)—‘VKG D’ and ‘VKG sweet’—in different soil matrices under natural climatic conditions. Dynamics of SFOs’ hydrocarbons (C10–C40), 16 PAHs, and a number of soil heterotrophic bacteria in oil-spiked soils was investigated during the long-term (1 year) outdoor experiment. In parallel, toxicity of aqueous leachates of oil-spiked soils to aquatic organisms (crustaceans Daphnia magna and Thamnocephalus platyurus and marine bacteria Vibrio fischeri) and terrestrial plants (Sinapis alba and Hordeum vulgare) was evaluated. Our data showed that in temperate climate conditions, the degradation of SFOs in the oil-contaminated soils was very slow: after 1 year of treatment, the decrease of total hydrocarbons’ content in the soil did not exceed 25 %. In spite of the comparable chemical composition of the two studied SFOs, the VKG sweet posed higher hazard to the environment than the heavier fraction (VKG D) due to its higher mobility in the soil as well as higher toxicity to aquatic and terrestrial species. Our study demonstrated that the correlation between chemical parameters (such as total hydrocarbons or total PAHs) widely used for the evaluation of the soil pollution levels and corresponding toxicity to aquatic and terrestrial organisms was weak.     

Oxidation of polycyclic aromatic hydrocarbons by horseradish peroxidase in water containing an organic cosolvent

Polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants that are toxic, mutagenic, and carcinogenic. We investigated the horseradish peroxidase (HRP)-catalyzed oxidation of PAHs in water containing N,N-dimethylformamide. Four PAHs (anthracene, phenanthrene, pyrene, and fluoranthene) were investigated using single-PAH and mixed-PAH systems. The results provide useful information regarding the preferential oxidation of anthracene over other PAHs regardless of the reaction time, enzyme dosage, and hydrogen peroxide concentration. The removal of PAHs was found to be very strongly correlated with the ionization potential (IP), and much greater PAH oxidation was observed at a lower IP. The oxidation of anthracene was specifically pH- and temperature-dependent, with the optimal pH and temperature being 8.0 and 40 °C, respectively. The redox mediators 1-hydroxybenzotriazole and veratryl alcohol promoted the transformation of anthracene by HRP; 9,10-anthraquinone was the main product detected from the anthracene oxidation system. The results of this study not only provide a better understanding of the oxidation of PAHs by utilizing a plant biocatalyst, but also provide a theoretical basis for establishing the HRP-catalyzed treatment of PAH-contaminated wastewater.     

Distribution of PAHs and trace metals in urban stormwater sediments: combination of density fractionation,mineralogy and microanalysis

Sediment management from stormwater infiltration basins represents a real environmental and economic issue for stakeholders due to the pollution load and important tonnages of these by-products. To reduce the sediment volumes to treat, organic and metal micropollutant-bearing phases should be identified. A combination of density fractionation procedure and microanalysis techniques was used to evaluate the distribution of polycyclic aromatic hydrocarbons (PAHs) and trace metals (Cd, Cr, Cu, Ni, Pb, and Zn) within variable density fractions for three urban stormwater basin sediments. The results confirm that PAHs are found in the lightest fractions (d?d??3) whereas trace metals are equally distributed within the light, intermediary, and highest fractions (d?d?d?d?>?2.8 g cm^?3) and are mostly in the 2.3?d??3 fraction. The characterization of the five fractions by global analyses and microanalysis techniques (XRD and MEB-EDX) allowed us to identify pollutant-bearing phases. PAHs are bound to the organic matter (OM) and trace metals to OM, clays, carbonates and dense particles. Moreover, the microanalysis study underlines that OM is the main constituent responsible for the aggregation, particularly for microaggregation. In terms of sediment management, it was shown that density fractionation is not suitable for trace metals but could be adapted to separate PAH-enriched phases.     

Indication of airborne pollution by birch and spruce in the vicinity of copper smelter

The aim of the study was determination of air pollution impact of the copper smelter in Bor and its surroundings (Serbia) by assessing the suitability of birch (Betula pendula Roth.) and spruce (Picea abies L.) for the purposes of biomonitoring and comparing it with previously published data from the same study area. The concentrations of Cu, Zn, Pb and Mn in leaves/needles, branches, roots and soil were determined. Sampling was performed during 2009 in two zones with high load of air pollution due to copper mining and smelting activities, and one background zone. Metal accumulation and translocation was evaluated in terms of biological factors. In addition, plant enrichment factor was calculated. According to the results, plant foliage was not enriched through soil, which indicates absorption from the air, with both species acting as excluders of Cu, Pb, Zn and Mn. Leaves were more enriched with all the metals than needles, indicating a better response of birch to airborne pollution than spruce. Cluster analysis showed different level of pollution at the sites, while correlations between Cu and Pb obtained by Principal Component Analysis indicated their anthropogenic origin. Regarding previously published results, beside birch leaves, pine needles (which showed higher level of response to pollution compared to linden leaves) could be applied in air biomonitoring surveys near copper smelters.     

The asexual enchytraeid worm Cognettia sphagnetorum (Oligochaeta) has increased Cu resistance in polluted soil

We studied Cu resistance in the asexual (reproduction through fragmentation) enchytraeid worms (Cognettia sphagnetorum, Oligochaeta) originating from two sites: one uncontaminated, and another contaminated by heavy metals. Adult worms were smaller and population density was lower at the polluted site. However, adults from the contaminated site had better survival in Cu-contaminated soil, but lower survival as juveniles (fragments). As we do not know the genetic basis of Cu resistance of the worms, it may have been reached by acclimatization via induced Cu regulation. Because fragmentation is the only mode of reproduction, all phenotypic properties (including resistance) of a parental generation could be transferred to filial generations.     

Occurrence of 1,1′-dimethyl-4,4′-bipyridinium (Paraquat) in irrigated soil of the Lake Chad Basin,Niger

Increased use of agrochemical products to improve yields for irrigated crops in sub-Saharan Africa has been accompanied by a significant increase in the risk of environmental contamination. Detailed examples of the fate of pesticides after initial spreading on crop fields are scarce in tropical regions, where safe practices and related health risks are poorly understood by smallholder farmers. In the semi-arid environment of the Lake Chad Basin, SE Niger, both intrinsic properties of pesticides and extrinsic factors such as soil and climate helped to characterize processes leading to an accumulation of pesticides in soils. Analysis by HPLC-UV of a 6 m deep soil profile showed the presence of Paraquat at concentrations from 953?±?102 μg kg^?1 to 3083?±?175 μg kg^?1 at depths between 0.80 and 2.75 m below the land surface. Soil analysis revealed that up to approximately 15 % of the total soil matrix consists of smectites, a clay mineral capable of retaining cationic pesticides such as Paraquat, and a very low content of organic matter (<0.15 wt.% TOC). Paraquat could be stored and not bioavailable in a clayey barrier at approximately 2-m depth and therefore does not represent an immediate risk for populations or environment in this form. However, if the Paraquat application rate remains constant, the clayey barrier could reach a saturation limit within 150–200 years and 180–220 years if we consider a DT50 in soil of ~1,000 days (FAO). Consequently, it could lead to a deeper infiltration and so a pollution of groundwater. Such a scenario can represent a health risk for drinking water and for the Lake Chad, which is a major resource for this densely populated region of semi-arid Africa. Further analyses should focus on deeper layers and groundwater Paraquat contents to validate or invalidate the hypothesis of storage in this clay-rich layer.     

Diversity and characterization of culturable bacterial endophytes from Zea mays and their potential as plant growth-promoting agents in metal-degraded soils

In this study, we evaluated the phylogenetic diversity of culturable bacterial endophytes of Zea mays plants growing in an agricultural soil contaminated with Zn and Cd. Endophytic bacterial counts were determined in roots and shoots, and isolates were grouped by random amplified polymorphic DNA and identified by 16S ribosomal RNA (rRNA) gene sequencing. Endophytes were further characterized for the production of plant growth-promoting (PGP) substances, such as NH₃, siderophores, indol-3-acetic acid (IAA), hydrogen cyanide and extracellular enzymes, and for the capacity to solubilize phosphate. The endophytes producing higher amounts of IAA were screened for their tolerance to Zn and Cd and used as bioinoculants for maize seedlings grown in the Zn/Cd-contaminated soil. The counts of endophytes varied between plant tissues, being higher in roots (6.48 log₁₀ g^?1 fresh weight) when compared to shoots (5.77 log₁₀ g^?1 fresh weight). Phylogenetic analysis showed that endophytes belong to three major groups: α-Proteobacteria (31 %), γ-Proteobacteria (26 %) and Actinobacteria (26 %). Pseudomonas, Agrobacterium, Variovorax and Curtobacterium were among the most represented genera. Endophytes were well-adapted to high Zn/Cd concentrations (up to 300 mg Cd l^?1 and 1,000 mg Zn l^?1) and showed ability to produce several PGP traits. Strains Ochrobactrum haematophilum ZR 3-5, Acidovorax oryzae ZS 1-7, Frigoribacterium faeni ZS 3-5 and Pantoea allii ZS 3-6 increased root elongation and biomass of maize seedlings grown in soil contaminated with Cd and Zn. The endophytes isolated in this study have potential to be used in bioremediation/phytoremediation strategies.