Flow cytometric analysis of hepatopancreatic cells from Armadillidium vulgare highlights terrestrial isopods as efficient environmental bioindicators in ex vivo settings.

Several studies have reported the high bioindication capacity of Isopoda (Crustacea, Oniscidea), which is related to their important ability to accumulate contaminants, usefulness in soil ecotoxicology and bioindication activities. Any change in the isopod population, diversity and life cycle can indicate relevant pollution levels. The analysis of target tissues, such as the hepatopancreas, is another emerging approach (from a cytologic/histological level) to detect contaminant accumulation from different sources. In this study, tissue disaggregation procedures were optimised in the hepatopancreas, and flow cytometry (FC) was applied to detect cell viability and several cell functions. After disaggregation, two hepatopancreatic cell types, small (S) and big (B), were still recognisable: they differed in morphology and behaviour. The analyses were conducted for the first time on isopods from sites under different conditions of ecological disturbance through cytometric re-interpretation of ecological-environmental parameters. Significant differences in cell functional parameters were found, highlighting that isopod hepatopancreatic cells can be efficiently analysed by FC and represent standardisable, early biological indicators, tracing environmental-induced stress through cytologic/histologic analyses.

The circadian clock gene bmal1 is necessary for co-ordinated circatidal rhythms in the marine isopod Eurydice pulchra (Leach).

Circadian clocks in terrestrial animals are encoded by molecular feedback loops involving the negative regulators PERIOD, TIMELESS or CRYPTOCHROME2 and positive transcription factors CLOCK and BMAL1/CYCLE. The molecular basis of circatidal (~12.4 hour) or other lunar-mediated cycles (~15 day, ~29 day), widely expressed in coastal organisms, is unknown. Disrupting circadian clockworks does not appear to affect lunar-based rhythms in several organisms that inhabit the shoreline suggesting a molecular independence of the two cycles. Nevertheless, pharmacological inhibition of casein kinase 1 (CK1) that targets PERIOD stability in mammals and flies, affects both circadian and circatidal phenotypes in Eurydice pulchra (Ep), the speckled sea-louse. Here we show that these drug inhibitors of CK1 also affect the phosphorylation of EpCLK and EpBMAL1 and disrupt EpCLK-BMAL1-mediated transcription in Drosophila S2 cells, revealing a potential link between these two positive circadian regulators and circatidal behaviour. We therefore performed dsRNAi knockdown of Epbmal1 as well as the major negative regulator in Eurydice, Epcry2 in animals taken from the wild. Epcry2 and Epbmal1 knockdown disrupted Eurydice's circadian phenotypes of chromatophore dispersion, tim mRNA cycling and the circadian modulation of circatidal swimming, as expected. However, circatidal behaviour was particularly sensitive to Epbmal1 knockdown with consistent effects on the power, amplitude and rhythmicity of the circatidal swimming cycle. Thus, three Eurydice negative circadian regulators, EpCRY2, in addition to EpPER and EpTIM (from a previous study), do not appear to be required for the expression of robust circatidal behaviour, in contrast to the positive regulator EpBMAL1. We suggest a neurogenetic model whereby the positive circadian regulators EpBMAL1-CLK are shared between circadian and circatidal mechanisms in Eurydice but circatidal rhythms require a novel, as yet unknown negative regulator.

Chemosensory proteins as putative semiochemical carriers in the desert isopod Hemilepistus reaumurii.

The order Isopoda contains both aquatic and terrestrial species, among which Hemilepistus reaumurii, which lives in arid environments and is the most adapted to terrestrial life. Olfaction has been deeply investigated in insects while it has received very limited attention in other arthropods, particularly in terrestrial crustaceans. In insects, soluble proteins belonging to two main families, Odorant Binding Proteins (OBPs) and Chemosensory Proteins (CSPs), are contained in the olfactory sensillar lymph and are suggested to act as carriers of hydrophobic semiochemicals to or from membrane-bound olfactory receptors. Other protein families, namely Nieman-Pick type 2 (NPC2) and Lipocalins (LCNs) have been also reported as putative odorant carriers in insects and other arthropod clades. In this study, we have sequenced and analysed the transcriptomes of antennae and of the first pair of legs of H. reaumurii focusing on soluble olfactory proteins. Interestingly, we have found 13 genes encoding CSPs, whose sequences differ from those of the other arthropod clades, including non-isopod crustaceans, for the presence of two additional cysteine residues, besides the four conserved ones. Binding assays on two of these proteins showed strong affinities for fatty acids and long-chain unsaturated esters and aldehydes, putative semiochemicals for this species.

Proteomics of the haemolymph of the terrestrial crustacean Porcellio scaber reveals components of its innate immunity under baseline conditions.

The terrestrial crustacean Porcellio scaber is an established test organism in environmental research. We analysed the haemolymph proteome of P. scaber using a classical proteomic approach based on one-dimensional gel electrophoresis and tandem mass spectrometry. Using a publicly available protein database and our P. scaber transcriptome data, we have identified 76 proteins involved in cytoskeleton formation, protein degradation, vesicular transport, genetic information processing, detoxification, carbohydrate and lipid metabolism reflecting haemocyte metabolic activity, active intracellular transport, and intercellular communication. Compared with the data reported for other crustaceans, 28 of these P. scaber proteins have been linked to its immunity, among them hemocyanin, α-2-macroglobulin, phenoloxidase 3, superoxide dismutase, glutathione S-transferase, haemolymph clottable protein, and histones H4 and H2B. Our results thus provide a firm base for studying the innate immune response of P. scaber at the level of the haemolymph proteome. This knowledge is of particular importance in ecotoxicity studies with various environmental stressors where understanding physiological changes is important to reveal possible modes of action.

Chemical forms of cadmium in soil and its distribution in French marigold sub-cells in response to chelator GLDA.

The use of degradable chelating agents to facilitate phytoextraction is a promising low-cost method for the remediation of heavy metal-contaminated soils. However, there are few studies on how plants and soils respond to the chelating agents. In this study, the responses of French marigold (Tagetes patula L.) and soil cadmium (Cd) to the chelator tetrasodium glutamate (GLDA) was investigated in a 180 d field trial. Five GLDA treatments (0, 292.5, 585, 1170, and 2340 kg hm-2) were carried out in a Cd-contaminated soil (0.47 mg kg-1) under French marigold plantation. The results showed that the application of GLDA promoted the transformation of other forms of Cd in soil to exchangeable state, and the exchangeable Cd and Fe-Mn oxide bound state increased by 42.13% and 32.97% (p < 0.05), respectively. The cell wall Cd accumulations significantly increased 9.39% (p < 0.05) and the percentages of soluble fractions increased by 460.33% (p < 0.05). Furthermore, increases occurred in soil pH, as well as DOC and DTPA-Cd contents with increasing the total amount of GLDA. The composite application of GLDA (2340 kg hm-2) with French marigold reduced the total soil Cd content by 7.59% compared with the soil background. Altogether, results of this study suggested that the application of GLDA can effectively activate soil Cd and enhance the capability of French marigold for the remediation of Cd-contaminated soils.

A comprehensive evaluation of the potential of semiterrestrial isopods, Ligia exotica, as a new animal food.

The semiterrestrial isopod, Ligia exotica represents one of the oldest documented species introductions of marine organisms and is known as an intermediate form between marine and strictly terrestrial isopods. In order to explore the potential value of Ligia as an animal food source, this study focused on the growth rate under laboratory rearing conditions and conducted a detailed analysis of the overall nutrient content of the species in comparison to two other marine food media (krill and fish meal). Evaluation of the growth rate of juveniles suggests it is a relatively fast-growing species of the Ligiidae family. The essential amino acids content Ligia meal is the lowest amongst the three studied media but the proportion of flavor amino acids, and in particular taurine, was higher. The most restricted amino acids of isopod meal are methionine and cysteine. The significantly unbalanced amino acid composition of Ligia meal may affect the absorption and utilization by consumers. In terms of fatty acids, the total polyunsaturated fatty acids in the isopod is very low. A total of 12 vitamins were examined. The VK1, VE, VB2, VB3, VB5 content of isopod meal were significantly higher than those of krill meal and fish meal. Similarly, most of the 11 mineral elements are highest in the isopod meal. Ligia therefore offers potential as an alternative natural food source in animal given the growth rate under culture and the overall nutrient content. But Ligia collected in most of the field would be deemed unfit for human consumption because of the relatively low nutritional value and heavy metal content exceeding the provided standard. Further study is warranted to elucidate the biological characteristics of isopods and how its diet is reflected in its nutritional value to consumers.

Intracellular calcium phosphate deposits contribute to transcellular calcium transport within the hepatopancreas of Porcellio scaber.

Like in most Crustacea, the cuticle of terrestrial isopods is hardened by a calcareous mineral phase. This rigid cuticle is frequently shed during a process called moulting. To reduce calcium loss, Porcellio scaber eats the shed cuticle, the exuviae, and absorb the calcium from it through large tubular diverticula of the intestine, called the mid gut glands or hepatopancreas. After moulting the absorbed calcium should be transported immediately into the hemolymph from which it is used to rapidly mineralize the new cuticle. This suggests that the hepatopancreas epithelium transports calcium from the lumen to the hemolymph. We used TEM, energy-filtered TEM and electron-probe X-ray microanalysis to analyse the distribution of elevated calcium within the hepatopancreas cells of P. scaber. We used animals in the postmoult stage that have eaten their exuviae and, as a control, those that have not ingested the exuviae. To minimize calcium loss within the samples, we used high pressure frozen and freeze substituted samples and propane-1-3-diol as floatation medium for thin-sectioning. The results reveal intracellular dense deposits containing calcium, phosphorus and oxygen at the apical microvillus membrane, within the cytoplasm, attached to vesicles and to the basolateral membrane, as well as extracellular between cells and the basal lamina. Control animals were devoid of these deposits. The results indicate that calcium from the exuviae is absorbed and transported across the epithelium into the hemolymph. We propose that during transport, intracellular calcium is bound to phosphate avoiding toxic effects of high concentrations of ionized calcium.

The shutting down of the insulin pathway: a developmental window for Wolbachia load and feminization.

Using the isopod Armadillidium vulgare as a case study, we review the significance of the "bacterial dosage model", which connects the expression of the extended phenotype to the rise of the Wolbachia load. In isopods, the Insulin-like Androgenic Gland hormone (IAG) induces male differentiation: Wolbachia feminizes males through insulin resistance, presumably through defunct insulin receptors. This should prevent an autocrine development of the androgenic glands so that females differentiate instead: feminization should translate as IAG silencing and increased Wolbachia load in the same developmental window. In line with the autocrine model, uninfected males expressed IAG from the first larval stage on, long before the androgenic gland primordia begin to differentiate, and exponentially throughout development. In contrast in infected males, expression fully stopped at stage 4 (juvenile), when male differentiation begins. This co-occurred with the only significant rise in the Wolbachia load throughout the life-stages. Concurrently, the raw expression of the bacterial Secretion Systems co-increased, but they were not over-expressed relative to the number of bacteria. The isopod model leads to formulate the "bacterial dosage model" throughout extended phenotypes as the conjunction between bacterial load as the mode of action, timing of multiplication (pre/post-zygotic), and site of action (soma vs. germen).

Shedding Light on the Antimicrobial Peptide Arsenal of Terrestrial Isopods: Focus on Armadillidins, a New Crustacean AMP Family.

In crustaceans, antimicrobial peptides (AMPs) are clustered into four major groups according to their amino acid composition and structure: (1) single-domain peptides containing cysteine residues such as anti-lipopolysaccharide-factor (ALF), (2) multi-domain or chimeric AMPs such as crustins, (3) non-conventional AMPs, and (4) linear single-domain AMPs. The majority of AMPs has been described in commercially exploited crustaceans, particularly decapods living in aquatic environments (crab, shrimp, lobster, and crayfish). Here, we aimed at establishing the AMPs repertoire of terrestrial isopods (Oniscidea), an original suborder of crustaceans adapted to life outside of the aquatic environment. Using transcriptomic data from 21 species, we identified 110 ALF and 73 crustin sequences. We also characterized the full-length sequence of armadillidins from 17 species, similar to the AMP previously described in the terrestrial isopod Armadillidium vulgare. Furthermore, we tested the antimicrobial activity of three armadillidin peptides characterized from three distantly related species. This analysis revealed similar activity spectra against pathogens, despite extensive structural variation among the tested peptides. In addition to conventional crustacean AMPs, our work highlights armadillidins as a new and independent family of AMPs specific to the Oniscidea, thus opening new perspectives concerning the study of the immune system of terrestrial isopods.

Remediation of heavy metals and enhancement of fertilizing potential of a sewage sludge by the synergistic interaction of woodlice and earthworms.

Woodlice and the earthworm alone or in combination were used to improve physical properties, nutrient release, and heavy metals stabilization during composting of sewage sludge. Chemical properties of raw sludge (IS) were compared to those of composted sludge (CS), sludge + earthworms (VS), sludge + woodlice (WS), and sludge + earthworms + woodlice (VWS) after 50 and 100 days of composting. Physical properties and heavy metals accumulation by the studied fauna was determined after 100 days of composting. Highest proportions of fine particles, porosity and water holding capacity were in the VWS treatment, electrical conductivity, ash content, inorganic N and the total concentrations of P, K, Ca and Mg increased, whereas the pH value, the organic C, C:N ratio and humic acids content decreased in following order: VWS > VC > WS > CS > IS. Total concentrations of Pb, Cd and Ni in composted sludge were lower whereas concentrations of Zn, Cu and Mn were higher than in raw sludge, and woodlice showed higher bioconcentration factor (BCFs) than earthworm for all heavy metals in all treatments. We concluded that woodlice and earthworms synergistically improved the physicochemical properties of the compost and enhanced its potential use as amendment in agricultural soils.

Comparative Tandem Mass Tag-Based Quantitative Proteomic Analysis of Tachaea chinensis Isopod During Parasitism.

Parasitic isopods perforate and attach to the host integument via the mandibles and then feed on hemolymph and exudate from the wounds. Such isopods attack a variety of commercially important fish and crustacean hosts. Similar to other hematophagous parasites, isopods may also employ biomolecules that affect host blood conglutination and defense systems. In the present study, a tandem mass tag-based quantitative proteomic approach was used to identify differentially expressed proteins in Tachaea chinensis parasites of shrimp, by comparing parasitic (fed) and pre-parasitic (unfed) individuals. We identified 888 proteins from a total of 1,510 peptides, with a significant difference in 129 between the fed and unfed groups. Among these, 37 were upregulated and 92 were downregulated in unfed T. chinensis. This indicates that T. chinensis may require more energy before parasitism during its search for a host. In addition, as is the case for other blood-sucking parasites, it might secrete antihemostatic, anti-inflammatory, and immunomodulatory molecules to facilitate blood meal acquisition. To our knowledge, this study is the first to use a TMT-based proteomic approach to analyze the proteome of isopod parasites, and the results will facilitate our understanding of the molecular mechanisms of isopod parasitism on crustaceans.

The effect of exuviae ingestion on lysosomal calcium accumulation and the presence of exosomes in the hepatopancreas of Porcellio scaber.

The hepatopancreas of isopods has major functions in food digestion and storage of carbohydrates and lipids. Also, it stores essential and accumulates xenobiotic metals in lysosomal granules within the two major cell types, the S- and B-cells of the tissue. A µCT study on moulting Porcellio scaber has shown mineral within the hepatopancreas lumen, when the animal has ingested their shed cuticle after moulting, suggesting recycling of mineral from the exuviae. This study aims to reveal if the lysosomal metal containing granules store calcium originating from the ingested exuviae. Therefore, we investigated the effect of cuticle ingestion on the elemental composition of the hepatopancreas granules of P. scaber, using electron probe X-ray microanalysis. For the preservation of diffusible elements, samples were high pressure frozen and freeze substituted in acetone and we used Propane-1,3-diol as a floatation medium for sections. We analyzed S- and B-cells of animals in the postmoult and intermoult stage that have ingested their exuviae and, as a negative control, cells from postmoult animals that have not ingested their exuviae. STEM and TEM were used for the investigation of the ultrastructure. Unexpectedly, the cryo-fixed samples contain numerous extracellular vesicles (exosomes) and many multivesicular bodies containing pro-exosomes. We show a significant increase of calcium, copper, zinc and sulphur within the metal granules upon exuviae ingestion, and, after 9 days, a reduction of calcium and zinc. The results indicate transitory storage of calcium from the exuviae within the metal granules and its subsequent utilization in cuticle mineralization.

Hypoxia causes woodlice (Porcellio scaber) to select lower temperatures and impairs their thermal performance and heat tolerance.

Environmental temperatures and oxygen availability are important for the balance between oxygen supply and demand. Terrestrial organisms are generally perceived to be less limited by access to oxygen than their aquatic counterparts. Nevertheless, even terrestrial environments can be deficient in oxygen, especially for organisms occurring in soil, litter, wood, rotten fruit or at high elevations. While isopods are the best adapted to a terrestrial lifestyle among crustaceans, many species, including woodlice, occupy environmental gradients of temperature and oxygen. To investigate whether mismatches between oxygen supply and demand can result in a loss of performance in a terrestrial organism, we studied the effects of atmospheric oxygen concentration on the thermal performance of the common rough woodlouse (Porcellio scaber). We compared the thermal preference, thermal sensitivity of running speed, and tolerance to extreme temperatures of woodlice exposed to one of two oxygen concentrations (21% - normoxia, 7% - hypoxia). Under hypoxia, P. scaber preferred microhabitats with temperatures that were on average 3°C lower than those preferred under normoxia. The running speed tended to reach its maximum at a lower temperature under hypoxia than under normoxia (25.13°C vs 28.87°C, respectively, although p was equal to 0.09), and normoxic woodlice ran approximately 1.5-fold faster than hypoxic woodlice at the point of maximum speed. Heat tolerance was significantly lower under hypoxia (38.9°C) than under normoxia (40.7°C), but there was no difference in cold tolerance (5.81°C under normoxia and 5.44°C under hypoxia). Overall, our results indicate that environmental gradients of temperature and oxygen may shape the physiological performance of terrestrial ectotherms, likely via their effects on the balance between oxygen supply and demand, which may have fitness consequences for these organisms in nature.

Medium-term dynamics of element concentrations in a sparid fish and its isopod parasite after the Prestige oil-spill: Shifting baselines?

Historically, the European Atlantic is probably the most important oil-spill hotspot worldwide. One of the most recent accidents occurred in 2002 when the oil-tanker Prestige sank over the Galician Bank causing two major oil-spills followed by several small leaks until March 2003. This resulted in contamination of virtually all types of marine habitat. Considering that parasites have proved to be good effect and accumulation bioindicators, the present study addresses the medium-term changes in trace element content after the Prestige oil-spill in a model host-parasite system, the bogue, Boops boops (Sparidae) and the isopod Ceratothoa oestroides. To our knowledge, this study is the first to address trace element concentrations in natural fish and parasite populations associated with the effects of an oil-spill. We observed that both test organisms examined, the host and the parasite, indicate a detectable change in the relative composition of trace element concentrations before and after the Prestige oil-spill. Multivariate analyses also indicated a differential response of the different tissues to the temporal sampling sequence. However, analyses of both host and parasite tissues supported the pattern of a gradual temporal transition to a state of relative trace element content distinctly departing from the pre-spill situation. Moreover, the parasite-host element accumulation ratios better depicted this temporal pattern. Additionally, changes in V concentrations in fish liver tissues and Ni concentrations in the parasite tissues suggest that this host-parasite system may be a useful tool to assess these two element contaminations linked to heavy fuel oil-spill.

Influence of Body Weight and Gender on the Accumulation of Heavy Metals in Ligia cinerascens Based on Field Investigation and Cd Exposure Experiment.

Isopods of the genus Ligia are deemed as excellent pollution monitoring organisms for coastal zones. A species of Ligia cinerascens was chosen to be tested, which was collected from three coastal zones in Dalian, China. The concentrations of metals followed the order of Cu>As ≈ Cd>Pb. Ligia collected from a municipal sewage outlet in Heishijiao showed the highest level of metals. Ligia from a natural bathing beach in Changshan Island showed the lowest level of metals as expected. Ligia grown in the same site could accumulate different levels of metals due to their different body weight. Both males and females conformably accumulated heavy metals in bodies. Taken together, this work suggests that L. cinerascens can serve as a biomonitor species. Within the range of 0.3-0.9 g body weights, body weight rather than gender should be taken into account as a factor in the process of environmental biomonitoring.

Stability and uptake of methylphenidate and ritalinic acid in nine-spine stickleback (Pungitius pungitius) and water louse (Asellus aquaticus).

The presence of human pharmaceuticals in the environment has garnered significant research attention because these compounds may exert therapeutic effects on exposed wildlife. Yet, for many compounds, there is still little research documenting their stability in the water column and uptake in organism tissues. Here, we measured the uptake and stability of methylphenidate (Ritalin®, a frequently prescribed central nervous system stimulant) and its primary metabolite, ritalinic acid, in (1) water only or (2) with nine-spine stickleback and water louse. Methylphenidate degraded to ritalinic acid in both studies faster at a higher temperature (20 °C versus 10 °C), with concentrations of ritalinic acid surpassing methylphenidate after 48-100 h, depending on temperature. The concentration of methylphenidate in stickleback was highest at the first sampling point (60 min), while the concentration in water louse tissues reached comparatively higher levels and peaked after ~ 6 days. Neither stickleback nor water louse took up ritalinic acid in tissues despite being present in the water column. Our findings provide valuable data for use in future risk assessment of methylphenidate and will aid in the design of studies aimed at measuring any ecotoxicological effects on, for example, the behaviour or physiology of aquatic organisms.

IGFBP-rP1, a strongly conserved member of the androgenic hormone signalling pathway in Isopoda.

The first protein which has been described to interact with the malacostracan Androgenic Gland Hormone (AGH) is a binding protein called IGFBP-rP1. It has been identified and studied in several species of decapods, in which its interaction with the masculinizing hormone and its expression patterns have been established in several ways. However, this protein remains uncharacterised to date in the other malacostracan orders, like Amphipoda and Isopoda, although they were historically the first ones in which the androgenic gland and the corresponding hormone were respectively described. In this article, we identified the IGFBP-rP1 of isopods and established its implication in the pathway of the AGH with a silencing approach in the model species Armadillidium vulgare. We also showed that this gene is expressed in all the tissues of males and females, with a similar pattern in animals infected with Wolbachia, a feminizing endosymbiont of several isopod species. The expression pattern did not differ during the development of uninfected and infected animals either. We finally studied the evolution of the IGFBP-rP1 in 68 isopod species, looking for conserved motifs and evidence of natural selection. Altogether, our results showed that this gene is constitutively expressed and strongly conserved in isopods, in which it likely constitutes a key element of the insulin/IGF signalling pathway. However, we also illustrated that IGFBP-rP1 is not sufficient on its own to explain the different developmental paths taken by the males and the females or feminized genetic males.

The in vivo effects of silver nanoparticles on terrestrial isopods, Porcellio scaber, depend on a dynamic interplay between shape, size and nanoparticle dissolution properties.

The present work aims to study the effects that acute exposure to low concentrations of silver nanoparticles (AgNPs) cause in digestive glands of terrestrial isopods (Porcellio scaber). The experiments were designed to integrate different analytical techniques, such as transmission electron microscopy, atomic absorption spectroscopy, proton induced X-ray emission, and Fourier transform IR imaging (FTIRI), in order to gain a comprehensive insight into the process from the AgNPs' synthesis to their interaction with biological tissues in vivo. To this aim, terrestrial isopods were fed with AgNPs having different shapes, sizes, and concentrations. For all the tested conditions, no toxicity at the whole organism level was observed after 14 days of exposure. However, FTIRI showed that AgNPs caused detectable local changes in proteins, lipids, nucleic acids and carbohydrates at the tissue level, to an extent dependent on the interplay of the AgNPs' properties: shape, size, concentration and dissolution of ions from them.

How does the bopyrid isopod Gyge branchialis interfere with trace metal bioaccumulation in the mud shrimp Upogebia cf. pusilla?

Parasites are widespread in natural environments, and their impacts on the fitness of their host and, at a broader scale, on ecosystem functioning are well recognized. Over the last two decades, there has been an increasing interest in the effects of parasites in conjunction with other stressors, especially pollutants, on the health of organisms. For instance, parasites can interfere with the bioaccumulation process of contaminants in their host leading to parasitized organisms exhibiting lower pollutants burdens than unparasitized individuals for example. However, the mechanisms underlying these patterns are not well understood. This study examined how the bopyrid parasite Gyge branchialis could lower the cadmium (Cd) uptake of its mud shrimp host Upogebia cf. pusilla. When exposed to water-borne Cd, parasites were able to bioaccumulate this trace metal. However, the uptake of Cd by the parasite was low and cannot entirely explain the deficit of Cd contamination of the host. The weight of gills of parasitized organisms was significantly reduced compared with unparasitized organisms. We suggest that by reducing the surface for metal uptake, parasites could lower the contaminant burden of their host.

The Effect of Urbanization on Trace Element Concentration and Symmetry of Woodlice (Armadillidium vulgare Latreille, 1804).

Woodlice are top consumers of a three-trophic system (soil, leaf litter, woodlice), and they are closely related to pollutant absorbing surfaces such as soil, leaf litter, and organic matter. We studied the effects of urbanization on trace element concentrations and fluctuating asymmetry of Armadillidium vulgare (Crustacea: Isopoda) individuals in and around Debrecen city, Hungary. Along an urbanization gradient (urban, suburban, and rural areas), trace element concentrations (Ba, Cu, Fe, Mn, Pb, and Zn) of woodlice and bioconcentration factor (BCF) were analyzed. Asymmetry was also measured in metric traits of woodlice: the length of three segments of antennae, the body length, and the width of the 3rd segment of pereon. We found significant differences in Ba and Cu concentration of A. vulgare individuals along the urbanization gradient. The highest Cu concentration was found in woodlice from the urban area and the highest Ba concentration was found in the individuals from the rural area. The Ba concentration was higher in females than in males. The BCF values of Cu indicated that A. vulgare accumulated this element from soil and leaf litter. There was no significant difference in symmetry of the bilateral traits of woodlice along the urbanization gradient based on FA levels. Our results showed that the urbanization had remarkable effect on the Ba and Cu concentration of woodlice which were originated from traffic pollution. At the same time, the anthropogenic activities did not affect the symmetry of the tested traits of A. vulgare individuals.