Data on the identification of microsatellite markers in Eisenia fetida and Eisenia andrei.

Eisenia fetida and Eisenia andrei are closely related earthworm species that play a crucial part in soil and influence its structure and organic matter cycling. Due to their essential environmental role, they are widely used as model organisms in a vast spectrum of research areas. In this work, we partially sequenced genomes of E. fetida and E. andrei, using Illumina technology (Nano 2 × 250 v2 - MiSeq) and de novo assembly strategy. A total of 3785 and 4258 microsatellite or Simple Sequence Repeat (SSR) markers were identified within E. fetida and E. andrei genomic DNA, respectively. The microsatellite markers will facilitate the analyses of genetic diversity and population genetics studies for the two selected earthworm species and their interspecific hybrids.

Air Pollution, Oxidative Stress, and the Risk of Development of Type 1 Diabetes.

Despite multiple studies focusing on environmental factors conducive to the development of type 1 diabetes mellitus (T1DM), knowledge about the involvement of long-term exposure to air pollution seems insufficient. The main focus of epidemiological studies is placed on the relationship between exposure to various concentrations of particulate matter (PM): PM1, PM2.5, PM10, and sulfur dioxide (SO2), nitrogen dioxide (NO2), carbon monoxide (O3), versus the risk of T1DM development. Although the specific molecular mechanism(s) behind the link between increased air pollution exposure and a higher risk of diabetes and metabolic dysfunction is yet unknown, available data indicate air pollution-induced inflammation and oxidative stress as a significant pathway. The purpose of this paper is to assess recent research examining the association between inhalation exposure to PM and associated metals and the increasing rates of T1DM worldwide. The development of modern and more adequate methods for air quality monitoring is also introduced. A particular emphasis on microsensors, mobile and autonomous measuring platforms, satellites, and innovative approaches of IoT, 5G connections, and Block chain technologies are also presented. Reputable databases, including PubMed, Scopus, and Web of Science, were used to search for relevant literature. Eligibility criteria involved recent publication years, particularly publications within the last five years (except for papers presenting a certain novelty or mechanism for the first time). Population, toxicological and epidemiological studies that focused particularly on fine and ultra-fine PM and associated ambient metals, were preferred, as well as full-text publications.

Transcriptome sequencing of Brassica napus highlights the complex issues with soil supplementation with sewage sludge.

The soil supplementation with sewage sludge (SS) has become a widespread method to improve soil quality, but its long-term possible consequences are still relatively unknown. SS may contain several groups of contaminants to which the biological responses of the organisms are still poorly understood mainly due to the mixture toxicity. In this context, RNA-seq has been used to assess the impact of the exposure to sewage sludge supplemented soil at the whole-transcriptome level in the Brassica napus (B. napus). Although the municipal sewage sludge passed all safety regulations set by the EU commission (86/278/EEC), soil supplementation with SS caused a significant (p < 0.05) increase in the content of lead (by 68.8%, 71.4% in plant shoots and roots, respectively), zinc (by 22.4% and 31.2%), nickel (by 67.0% and 30.2%), and copper (by 33.1% and 39.2%). The de-novo assembled transcriptome of B. napus identified 555 differently expressed genes (DEGs) in a response to sewage sludge supplementation at the false detection rate below 0.001 (FDR <0.001). Among them, 313 genes were up-regulated and 242 genes were down-regulated. The gene ontology analysis (GO) had shown, that significantly enriched GO groups included genes involved in photosynthesis, carbohydrate metabolism and photosystems repair (41 genes), response to oxidative stress (50 genes), response to pathogens (36 genes), response to xenobiotics (15 genes), and heavy metals (41 genes), cell death (8 genes), cell wall structure (15 genes). These results suggest a significant impact of contaminants in sewage sludge on plants transcriptome. The transcriptomic approach facilitated a better understanding of the molecular level of the potential toxicity of sewage sludge in B. napus. RNA-seq allowed for the identification of potential novel early-warning molecular markers of environmental contamination. This work highlights the crucial necessity for rapid legislation change concerning the allowable levels of contaminants in sewage sludge applied on land, to mitigate the possible adverse outcomes in the ecosystem after its use as a fertilizer.

Polymorphic microsatellite markers demonstrate hybridization and interspecific gene flow between lumbricid earthworm species, Eisenia andrei and E. fetida.

The lumbricid earthworms Eisenia andrei (Ea) and E. fetida (Ef) have been used as model organisms for studies on hybridization. Previously they have been identified by species specific sequences of the mitochondrial COI gene of maternal origin ('a' or 'f') and the nuclear 28S gene of maternal/paternal origin ('A' or 'F'). In experimental crosses, these hermaphroditic species produce progeny of genotypes Ea (aAA), Ef (fFF) and hybrids (aAF and fFA) originating by self-fertilization or cross-fertilization. To facilitate studies on new aspects of the breeding biology and hybridization of earthworms, polymorphic microsatellite markers were developed based on 12 Ea and 12 Ef specimens and validated on DNA samples extracted from 24 genotyped specimens (aAA, fFF, aAF and fFA) from three laboratory-raised families and 10 of them were applied in the present study. The results indicate that microsatellite markers are valuable tools for tracking interspecific gene flow between these species.

Transfer of sulfidized silver from silver nanoparticles, in sewage sludge, to plants and primary consumers in agricultural soil environment.

Consumer products containing silver nanoparticles (AgNPs) release silver (Ag) to the environment, particularly wastewater. Sewage sludge (SS), which contains numerous contaminants including Ag, is recycled by spreading on agricultural land. Although slight impacts and bioaccumulation of Ag sulfide (Ag2S, the main species found in SS) in terrestrial organisms have been demonstrated, possible trophic transfer into plants and subsequently animal species has not been examined. Accordingly, the present study experimentally measured the transfer of Ag from AgNPs and sulfidized Ag into plants and primary consumers and compared their bioavailability. Nine plant cultivars were grown in soil mixed with SS containing Ag, which revealed that bioaccumulation of Ag by plants is species-dependent. Ryegrass (the plant species with the greatest accumulation - up to 0.2 mg kg-1) was then cultivated on a larger scale to expose snails and locusts for several weeks. While locusts did not accumulate Ag after two weeks of exposure, snails exhibited Ag bioaccumulation after 5 weeks when soil was accessible. Sulfidized Ag derived from AgNPs were less available (bioaccumulation up to 2.5 mg kg-1) than the Ag from the original AgNPs (bioaccumulation up to 15 mg kg-1). This transfer potential of Ag could have consequences for food webs due to chronic exposure linked to SS spreading practices. This study shows that transformations of AgNPs in treatment plants attenuate but do not completely eliminate the risk of Ag to plant and animal species SS.

In-site and Ex-site Date Palm Exposure to Heavy Metals Involved Infra-Individual Biomarkers Upregulation.

As a tree of considerable importance in arid regions-date palm, Phoenix dactylifera L. survival in contaminated areas of Sfax city has drawn our attention. Leaf samples of the plants grown in the study area showed high levels of cadmium (Cd), copper (Cu), and chromium (Cr). On the basis of this finding, the cellular mechanisms that explain these metal accumulations were investigated in controlled conditions. After four months of exposure to Cd, Cr, or Cu, high bioconcentration and translocation factor (TF>1) have been shown for date palm plantlets exposed to Cd and low TF values were obtained for plantlets treated with Cr and Cu. Moreover, accumulation of oxidants and antioxidant enzyme activities occurred in exposed roots to Cu and Cd. Secondary metabolites, such as polyphenols and flavonoids, were enhanced in plants exposed at low metal concentrations and declined thereafter. Accumulation of flavonoids in cells may be correlated with the expression of the gene encoding Pdmate5, responsible for the transport of secondary metabolites, especially flavonoids. Other transporter genes responded positively to metal incorporation, especially Pdhma2, but also Pdabcc and Pdnramp6. The latter would be a new candidate gene sensitive to metallic stress in plants. Expressions of gene coding metal chelators were also investigated. Pdpcs1 and Pdmt3 exhibited a strong induction in plants exposed to Cr. These modifications of the expression of some biochemical and molecular based-markers in date palm helped to better understand the ability of the plant to tolerate metals. They could be useful in assessing heavy metal contaminations in polluted soils and may improve accumulation capacity of other plants.

Impact of cadmium on forage kale (Brassica oleracea var. viridis cv "Prover") after 3-,10- and 56-day exposure to a Cd-spiked field soil.

Cadmium (Cd) is a highly toxic element for living organisms and is widespread in metal-contaminated soils. As organisms which can grow up on these polluted areas, plants have some protection mechanisms against Cd issues. Among the plant kingdom, the Brassicaceae family includes species which are known to be able to tolerate and accumulate Cd in their tissues. In this study, Brassica oleracea var. viridis cv "Prover" was exposed to a range of artificially Cd-contaminated soils (from 2.5 up to 20 mg kg-1) during 3, 10, and 56 days and the effects on life traits, photosynthesis activity, antioxidant enzymatic activities were studied. Metal accumulation was quantified, as well as DNA damage, by means of the comet assay and immunodetection of 8-OHdG levels. Globally, B. oleracea was relatively tolerant to those Cd exposures. However, comet assay and detection of 8-OHdG revealed some DNA damage but which are not significant. According to metal accumulation analysis, B. oleracea var. viridis cv Prover could be a good candidate for alternative growing in contaminated areas.

Accumulation, speciation and localization of silver nanoparticles in the earthworm Eisenia fetida.

The use of silver nanoparticles (AgNPs) in agriculture and many consumer products has led to a significant release of Ag in the environment. Although Ag toxicity in terrestrial organisms has been studied extensively, very little is known about the accumulation capacity and coping mechanisms of organisms in Ag-contaminated soil. In this context, we exposed Eisenia fetida earthworms to artificial OECD soil spiked with a range of concentrations of Ag (AgNPs or AgNO3). The main aims were to (1) identify the location and form of accumulation of Ag in the exposed earthworms and (2) better understand the physiological mechanisms involved in Ag detoxification. The results showed that similar doses of AgNPs or AgNO3 did not have the same effect on E. fetida survival. The two forms of Ag added to soil exhibited substantial differences in speciation at the end of exposure, but the Ag speciation and content of Ag in earthworms were similar, suggesting that biotransformation of Ag occurred. Finally, 3D images of intact earthworms obtained by X-ray micro-computed tomography revealed that Ag accumulated preferentially in the chloragogen tissue, coelomocytes, and nephridial epithelium. Thus, E. fetida bioaccumulates Ag, but a regulation mechanism limits its impact in a very efficient manner. The location of Ag in the organism, the competition between Ag and Cu, and the speciation of internal Ag suggest a link between Ag and the thiol-rich proteins that are widely present in these tissues, most probably metallothioneins, which are key proteins in the sequestration and detoxification of metals.

Medium-term effects of Ag supplied directly or via sewage sludge to an agricultural soil on Eisenia fetida earthworm and soil microbial communities.

The widespread use of silver nanoparticles (AgNPs) in consumer products that release Ag throughout their life cycle has raised potential environmental concerns. AgNPs primarily accumulate in soil through the spreading of sewage sludge (SS). In this study, the effects of direct exposure to AgNPs or indirect exposure via SS contaminated with AgNPs on the earthworm Eisenia fetida and soil microbial communities were compared, through 3 scenarios offering increasing exposure concentrations. The effects of Ag speciation were analyzed by spiking SS with AgNPs or AgNO3 before application to soil. SS treatment strongly impacted Ag speciation due to the formation of Ag2S species that remained sulfided after mixing in the soil. The life traits and expression of lysenin, superoxide dismutase, cd-metallothionein genes in earthworms were not impacted by Ag after 5 weeks of exposure, but direct exposure to Ag without SS led to bioaccumulation of Ag, suggesting transfer in the food chain. Ag exposure led to a decrease in potential carbon respiration only when directly added to the soil. The addition of SS had a greater effect on soil microbial diversity than the form of Ag, and the formation of Ag sulfides in SS reduced the impact of AgNPs on E. fetida and soil microorganisms compared with direct addition.

Species-specific Cd-detoxification mechanisms in lumbricid earthworms Eisenia andrei, Eisenia fetida and their hybrids.

Hermaphroditic lumbricid Eisenia sp. earthworms are ubiquitous and highly resistant to a variety of environmental stressors, including heavy metals. Among the progeny of laboratory mated inter-specific pairs of Eisenia fetida (Ea) and Eisenia andrei (Ef) there are fertile Ha hybrids derived from Ea ova fertilized by Ef spermatozoa and very rare sterile Hf hybrids from Ef ova fertilized by Ea spermatozoa. The aim of the first part of the experiment was to compare the life traits and whole body accumulation of cadmium in adult earthworms from genetically defined Ea, Ef and their hybrids (Ha) exposed for four weeks to commercial soil either unpolluted (control) or cadmium-spiked leading to moderate (M) or high (H) soil pollution (M = 425 and H = 835 mg kg-1 dry soil weight). Such exposure impaired cocoon production but not affected earthworm viability despite the massive Cd bioaccumulation in the whole earthworm bodies reaching at M and H groups 316-454, 203-338, 114-253, and 377-309 mg kg-1 dry body weights of Ea, Ef1, Ef2, and Ha, respectively, surprisingly reaching maximum accumulation quantities in hybrids. The second part of the experiment aimed to investigate cadmium-related defense mechanisms at transcriptomic level in coelomocytes non-invasively extruded from coelomic cavities of the new sets of Ea, Ef, Ha, and Hf earthworms exposed to Cd in microcosms for 0 days (control), 2 days, and 7 days (M = 425 mg kg-1). Expression level of stress-induced Cd-metallothionein (mt) and superoxide dismutase (sod) were gradually up-regulated, while the immune-connected lysenin (lys) was rapidly down-regulated; the expression of glutathione S-transferase (gst) and phytochelatin synthase (pcs) remained unaffected. Mt and sod gene up-regulation and lys gene down-regulation were especially pronounced in Ea-derived hybrids. In sum, capacity of cadmium bioaccumulation and detoxification mechanisms is more efficient in interspecific hybrids than in the pure Ea and Ef species.

Modeling and optimizing the removal of cadmium by Sinapis alba L. from contaminated soil via Response Surface Methodology and Artificial Neural Networks during assisted phytoremediation with sewage sludge.

The study was aimed to model and optimize the removal of cadmium from contaminated post-industrial soil via Sinapis alba L. by comparing two modeling approaches: Response Surface Methodology (RSM) and Artificial Neural Networks (ANN). The experimental design was done using the Box-Behnken Design method. In the RSM model, the quadratic model was shown to predict the closest results in comparison to our experimental data. For ANN approach, a two-layer Feed-Forward Back-Propagation Neural Network model was designed. The results showed that sewage sludge supplementation increased the efficiency of the Sinapis alba plant in removing Cd from the soil. After 28 days of exposure, the removal rate varied from 10.96% without any supplementation to 65.9% after supplementation with the highest possible (law allowed) dose of sewage sludge. The comparison proved that the prediction capability of the ANN model was much higher than that of the RSM model (adjusted R-square: 0.98, standard error of the Cd prediction removal: 0.85 ± 0.02). Thus, the ANN model could be used for the prediction of heavy metal removal during assisted phytoremediation with sewage sludge. Moreover, such approach could also be used to determinate the dose of sewage sludge that will ensure highest process efficiency.

Effects of sewage sludge supplementation on heavy metal accumulation and the expression of ABC transporters in Sinapis alba L. during assisted phytoremediation of contaminated sites.

ATP binding cassette (ABC) transporters, types C, G, and B were monitored via qPCR in order to investigate the influence of heavy metal (HM) contamination of post-industrial and post-agricultural soils and the effects of its supplementation with sewage sludge, on Sinapis alba plants. Five house-keeping genes were selected and validated to ensure the best reference points. The relative expression of ABC types C and G genes was profoundly affected by experimental conditions and included their upregulation after plants exposure to heavy metals and downregulation after supplementation with sewage sludge. However, ABC type C was more responsive then type G. The experimental conditions altered the expression of ABC type C gene faster than ABC type G and thus, the expression of ABC type C can therefore potentially be used as a bioindicator during assisted phytoremediation of degraded sites. In clean soil, supplementation with sewage sludge with a slight content of heavy metals still caused an upregulation in the expression of ABC types C and G, which showed that proper toxicity assessments are necessary to ensure safe application of sewage sludge into soils. Results showed that the analysed genes take a significant part in plants metal detoxification and that their expression is regulated at transcriptional level after exposure to soil contaminated with heavy metals by both, industrial activities and by sewage sludge supplementation. Thus, their expression can potentially be used as an early-warning biomarker when soil supplementation with sewage sludge is incorporated into the soil-management process.

Impacts of metallic trace elements on an earthworm community in an urban wasteland: Emphasis on the bioaccumulation and genetic characteristics in Lumbricus castaneus.

Metallic trace elements (MTEs) soil pollution has become a worldwide concern, particularly regarding its impact on earthworms. Earthworms, which constitute the dominant taxon of soil macrofauna in temperate regions and are crucial ecosystem engineers, are in direct contact with MTEs. The impacts of MTE exposure on earthworms, however, vary by species, with some able to cope with high levels of contamination. We combined different approaches to study the effects of MTEs at different levels of biological organisation of an earthworm community, in a contaminated urban wasteland. Our work is based on field collection of soil and earthworm samples, with a total of 891 adult earthworms from 8 species collected, over 87 quadrats across the study plot. We found that MTE concentrations are highly structured at the plot scale and that some elements, such as Pb, Zn, and Cu, are highly correlated. Comparing species assemblage to MTE concentrations, we found that the juvenile and adult abundances, and community composition, were significantly affected by pollution. Along the pollution gradient, as species richness decreased, Lumbricus castaneus became more dominant. We thus investigated the physiological response of this species to a set of specific elements (Pb, Zn, Cu, and Cd) and studied the impacts of MTE concentrations at the plot scale on its population genetic. These analyses revealed that L. castaneus is able to bioaccumulate high quantities of Cd and Zn, but not of Cu and Pb. The population genetic analysis, based on the genotyping of 175 individuals using 8 microsatellite markers, provided no evidence of the role of the heterogeneity in MTE concentrations as a barrier to gene flow. The multidisciplinary approach we used enabled us to reveal the comparatively high tolerance of L. castaneus to MTE concentrations, suggesting that this is a promising model to study the molecular bases of MTE tolerance.

Modelling assisted phytoremediation of soils contaminated with heavy metals - Main opportunities, limitations, decision making and future prospects.

The heavy metals (HMs) soils contamination is a growing concern since HMs are not biodegradable and can accumulate in all living organisms causing a threat to plants and animals, including humans. Phytoremediation is a cost-efficient technology that uses plants to remove, transform or detoxify contaminants. In recent years, phytoremediation is entering the stage of large-scale modelling via various mathematical models. Such models can be useful tools to further our understanding and predicting of the processes that influence the efficiency of phytoremediation and to precisely plan such actions on a large-scale. When dealing with extremely complicated and challenging variables like the interactions between the climate, soil and plants, modelling before starting an operation can significantly reduce the time and cost of such process by granting us an accurate prediction of possible outcomes. Research on the applicability of different modelling approaches is ongoing and presented work compares and discusses available models in order to point out their specific strengths and weaknesses in given scenarios. The main aim of this paper is to critically evaluate the main advantages and limitations of available models for large-scale phytoremediation including, among others, the Decision Support System (DSS), Response Surface Methodology (RSM), BALANS, PLANTIX and various regression models. Study compares their applicability and highlight existing gaps in current knowledge with a special reference to improving the efficiency of large-scale phytoremediation of sites contaminated with heavy-metals. The presented work can serve as a useful tool when choosing the most suitable model for the phytoremediation of contaminated sites.

Ecotoxicology of silver nanoparticles and their derivatives introduced in soil with or without sewage sludge: A review of effects on microorganisms, plants and animals.

Silver nanoparticles (AgNPs) are widely incorporated in many products, partly due to their antimicrobial properties. The subsequent discharge of this form of silver into wastewater leads to an accumulation of silver species (AgNPs and derivatives resulting from their chemical transformation), in sewage sludge. As a result of the land application of sewage sludge for agricultural or remediation purposes, soils are the primary receiver media of silver contamination. Research on the long-term impact of AgNPs on the environment is ongoing, and this paper is the first review that summarizes the existing state of scientific knowledge on the potential impact of silver species introduced into the soil via sewage sludge, from microorganisms to earthworms and plants. Silver species can easily enter cells through biological membranes and affect the physiology of organisms, resulting in toxic effects. In soils, exposure to AgNPs may change microbial biomass and diversity, decrease plant growth and inhibit soil invertebrate reproduction. Physiological, biochemical and molecular effects have been documented in various soil organisms and microorganisms. Negative effects on organisms of the dominant form of silver in sewage sludge, silver sulfide (Ag2S), have been observed, although these effects are attenuated compared to the effects of metallic AgNPs. However, silver toxicity is complex to evaluate and much remains unknown about the ecotoxicology of silver species in soils, especially with respect to the possibility of transfer along the trophic chain via accumulation in plant and animal tissues. Critical points related to the hazards associated with the presence of silver species in the environment are described, and important issues concerning the ecotoxicity of sewage sludge applied to soil are discussed to highlight gaps in existing scientific knowledge and essential research directions for improving risk assessment.

Gene expression, DNA damage and other stress markers in Sinapis alba L. exposed to heavy metals with special reference to sewage sludge application on contaminated sites.

Bioindicators are promising tools used to detect the long-term effects of selected biosolids on plants development and should be implemented before large-scale supplementation of sewage sludge into the soil. The presented study shows the impact of sewage sludge application on metal-sensitive toxicity biological parameters (biomarkers) in Sinapis alba including: germination, root length, the activity of guaiacol peroxidase, the chlorophyll content, the level of DNA damage and the expression level of Ribulose-1,5-bisphosphate carboxylase/oxygenase (rbcL) and metallothionein (mt). We evaluated data from selected biomarkers in order to broaden our understanding of plants defense mechanisms against heavy metal contamination and the application of sewage sludge into soils. Overall, in contaminated soil after supplementation with both municipal sewage sludges, an increase in toxicity was noticed in DNA damage, mt and rbcl expression and total chlorophyll content. The supplementation of both soils with municipal sewage sludge caused a two-time induction in the mt expression. Moreover, clean soil supplemented with sewage sludge caused an increase in DNA damage shown as the tail moment from approximately 12 µm on control to 40 µm after supplementation. Even if those biosolids increased the initial germination, roots length, and biomass in comparison to the unamended soil, the toxicity was evidenced with other stress markers. Results showed, that in order to accurately assess the influence of sewage sludge application on plants the use of several specific biomarkers is required for safe land restoration. The conducted study also confirmed, both under biochemical and genotoxic tests, that iron enrichment for biosolids or contaminated soil can significantly reduce the bioavailability and toxicity of other metals.

Bioaccumulation, antioxidative response, and metallothionein expression in Lupinus luteus L. exposed to heavy metals and silver nanoparticles.

Yellow-lupin (Lupinus luteus L.) was grown on soils contaminated with heavy metals during two parallel studies. In the first one, the soil was contaminated by industrial activities whereas, in the second one, the soil was artificially contaminated with a single metal including Cd, Pb, Zn, Ni (in nitrate form), and Ag (in nitrate and nanoparticles form). The study was performed to assess a plant's response to contamination including its antioxidative response and molecular mechanisms involved in metal detoxification through the expression level of metallothioneins (MTs). Overall, the study provided insights into identification and validation of housekeeping genes (HKG) in L. luteus under exposure to metal stress and showed the effects of selected heavy metals and silver nanoparticles on the expression of metallothioneins, the activity of guaiacol peroxidase (GPX) and bioaccumulation of metals in leaves of L. luteus. As such, HKG validation using BestKeeper, NormFinder, and geNorm software allowed for the selection of four most stable reference genes in a context metal contamination for the selected plant. Moreover, a significant increase in the expression levels of MT was observed in plants grown under heavy metal stress and none on plants grown on 25 mg kg-1 of silver nanoparticles. Also, the GPX activity and MT expression showed statistically significant changes between different conditions and doses which means that they can be used as highly sensitive stress markers for planning the phytoremediation process on a large scale.

Insight into the expression variation of metal-responsive genes in the seedling of date palm (Phoenix dactylifera).

Phytochelatin synthase and metallothionein gene expressions were monitored via qPCR in order to investigate the molecular mechanisms involved in Cd and Cr detoxification in date palm (Phoenix dactylifera). A specific reference gene validation procedure using BestKeeper, NormFinder and geNorm programs allowed selection of the three most stable reference genes in a context of Cd or Cr contamination among six reference gene candidates, namely elongation factor α1, actin, aldehyde dehydrogenase, SAND family, tubulin 6 and TaTa box binding protein. Phytochelatin synthase (pcs) and metallothionein (mt) encoding gene expression were induced from the first days of exposure. At low Cd stress (0.02 mM), genes were still up-regulated until 60th day of exposure. At the highest metal concentrations, however, pcs and mt gene expressions decreased. pcs encoding gene was significantly up-regulated under Cr exposure, and was more responsive to increasing Cr concentration than mt encoding gene. Moreover, exposure to Cd or Cr influenced clearly seed germination and hypocotyls elongation. Thus, the results have proved that both analyzed genes participate in metal detoxification and their expression is regulated at transcriptional level in date palm subjected to Cr and Cd stress. Consequently, variations of expression of mt and pcs genes may serve as early-warning biomarkers of metal stress in this species.

The existence of fertile hybrids of closely related model earthworm species, Eisenia andrei and E. fetida.

Lumbricid earthworms Eisenia andrei (Ea) and E. fetida (Ef) are simultaneous hermaphrodites with reciprocal insemination capable of self-fertilization while the existence of hybridization of these two species was still debatable. During the present investigation fertile hybrids of Ea and Ef were detected. Virgin specimens of Ea and Ef were laboratory crossed (Ea+Ef) and their progeny was doubly identified. 1 -identified by species-specific maternally derived haploid mitochondrial DNA sequences of the COI gene being either 'a' for worms hatched from Ea ova or 'f' for worms hatched from Ef ova. 2 -identified by the diploid maternal/paternal nuclear DNA sequences of 28s rRNA gene being either 'AA' for Ea, 'FF' for Ef, or AF/FA for their hybrids derived either from the 'aA' or 'fF' ova, respectively. Among offspring of Ea+Ef pairs in F1 generation there were mainly aAA and fFF earthworms resulted from the facilitated self-fertilization and some aAF hybrids from aA ova but none fFA hybrids from fF ova. In F2 generation resulting from aAF hybrids mated with aAA a new generations of aAA and aAF hybrids were noticed, while aAF hybrids mated with fFF gave fFF and both aAF and fFA hybrids. Hybrids intercrossed together produced plenty of cocoons but no hatchlings independently whether aAF+aAF or aAF+fFA were mated. These results indicated that Ea and Ef species, easy to maintain in laboratory and commonly used as convenient models in biomedicine and ecotoxicology, may also serve in studies on molecular basis of interspecific barriers and mechanisms of introgression and speciation. Hypothetically, their asymmetrical hybridization can be modified by some external factors.

Copper toxicity and date palm (Phoenix dactylifera) seedling tolerance: Monitoring of related biomarkers.

Date palm (Phoenix dactylifera) seeds were exposed to different copper (Cu) solutions to examine plant stress responses. Low Cu concentrations (0.02 and 0.2 mM) caused an increase of seed germination, whereas higher Cu amounts (2 mM) significantly inhibited seed germination, delayed hypocotyl elongation, increased seedling mortality, and reduced the germination index by more than 90%. Metal-related toxicity symptoms appeared after 15 d of 2 mM of Cu exposure. Biochemical activities such as amylase activity and redox balance elements were examined to study the relationship between external Cu amount and internal plant response. The present study showed that amylolytic activity was dose- and time-dependent. Likewise, H2 O2 production increased after exposure to Cu, which was correlated with thiobarbituric acid reactive substance (TBARS) accumulation. Furthermore at low Cu concentrations, superoxide dismutase (SOD) and catalase (CAT) activities increased, suggesting that date palm seed stimulated its metal homeostasis networks. However, the highest cupric ion amounts increased cell oxidant accumulation and reduced enzyme production. Gene expression level measures of P. dactylifera phytochelatin synthase (Pdpcs) and P. dactylifera metallothionein (Pdmt) encoding genes have been carried out to investigate the implication of PdPCS and PdMT proteins in Cu homeostasis and/or its sequestration. Phoenix dactylifera metallothionein induction reached a peak after 30 d of exposure to 0.2 mM of Cu. However, it was down-regulated in plants exposed to higher Cu concentrations. In the same conditions, Pdpcs was overexpressed during 1 mo of exposure before it decreased thereafter. These observations provide a new insight into date palm cell response to Cu, a metal that can be toxic but that is also an essential element. Environ Toxicol Chem 2018;37:797-806. © 2017 SETAC.