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Inhibition of oxidative stress and ferroptosis is currently considered to be a promising therapeutic approach for neurodegenerative diseases. Herpotrichones, a class of compounds derived from insect symbionts, have shown potential for neuroprotective activity with low toxicity. However, the specific mechanisms through which herpotrichones exert their neuroprotective effects remain to be fully elucidated. In this study, the natural [4 + 2] adducts herpotrichone A ( He-A ) and its new analogues were isolated from the isopod-associated fungus Herpotrichia sp. SF09 and exhibited significantly protective effects in H 2 O 2 -, 6-OHDA-, and RSL3-stimulated PC12 cells and LPS-stimulated BV-2 cells. Moreover, He-A was able to relieve ferroptotic cell death in RSL3-stimulated PC12 cells and 6-OHDA-induced zebrafish larvae. Interestingly, He-A can activate antioxidant elements and modulate the SLC7A11 pathway without capturing oxidic free radical and chelating iron. These findings highlight He-A as a novel hit that protects against ferroptosis-like neuronal damage in the treatment of neurodegenerative diseases.

The exoskeletons of crustaceans are essential for providing protection from predators and other environmental threats. Understanding the structure and mechanical behavior of their natural armor could inspire the design of lightweight and high toughness synthetic materials. Most published work has focused on marine crustacea rather than their terrestrial counterparts, which are exposed to a multitude of unique threats. The interest in the terrestrial isopod Armadillidium vulgare (A. vulgare) has grown but the interrelationship between the microstructure, chemical composition, and mechanical properties has not been thoroughly investigated. Thus, this study aims to elucidate missing details concerning this biological mineralized composite. Exoskeleton specimens were fixated to preserve the intrinsic protein structure. We utilize scanning electron microscopy for microstructure analysis, Raman spectroscopy for elemental analysis, and nanoindentation property mapping to achieve mechanical characterization. The naturally fractured A. vulgare exoskeleton cross-section reveals four subregions with the repeating helicoidal 'Bouligand' arrangement most prominent in the endocuticle. The hardness and reduced modulus distributions exhibit a through-thickness exponential gradient with decreasing magnitudes from the outermost to the innermost layers of the exoskeleton. The Raman spectra show a graded spatial distribution of key constituents such as calcium carbonate across the thickness, some of which are consistent with the mechanical property gradient. Potential microstructure, elemental composition, and mechanical property relationships are discussed to explain how the hierarchical structure of this nanolaminate armor protects this species., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Wound healing is a fundamental response to tissue injury and a number of natural products has been found to accelerate the healing process. Herein, we report the preparation of a series of different polarity (organic and aqueous) extracts of the marine isopod Ceratothoa oestroides and the in vivo evaluation of their wound healing activity after topical administration of ointments incorporating the various extracts on wounds inflicted on SKH-hr1 hairless mice. The most active extract was fractionated for enrichment in the bioactive constituents and the fractions were further evaluated for their wound healing activity, while their chemical profiles were analyzed. Wound healing was evaluated by clinical assessment, photo-documentation, histopathological analysis and measurement of biophysical skin parameters, such as transepidermal water loss (TEWL), hydration, elasticity, and skin thickness. The highest levels of activity were exerted by treatment of the wounds with a fraction rich in eicosapentaenoic acid (EPA), as well as myristic and palmitoleic acids. Topical application of the bioactive fraction on the wounds of mice resulted in complete wound closure with a skin of almost normal architecture without any inflammatory elements.

To estimate trace element bioaccumulation in Armadillo officinalis, specimens were collected from Ghar El Melh lagoon then exposed for 3 weeks in contaminated sediments with copper, zinc, and cadmium. From the first week until the end of the experiment, a decrease in A. officinalis growth related to the increase of Cd concentration in the sediment was recorded. However, a mass gain was highlighted under Cu and Zn exposures. At the end of experiment, body metal concentrations were measured using flame atomic emission spectrometry. Results of the bioaccumulation factor showed that the species could be considered as a macroconcentrator of copper (BAF > 2) and a deconcentrator of zinc (BAF < 2). Microscopy observations of hepatopancreas cells showed morphological and histological changes even at the lowest concentration. They consisted in the microvillus border destruction, lipid droplets modifications, trace element accumulation, and the condensation of the majority of cellular organelles. The degree of these alterations was found to be dose-dependent. Through these results, the isopod A. officinalis could be used as relevant monitor organisms for soil metal contamination., (© 2019 Wiley Periodicals, Inc.)

This study was conducted to compare metals bioaccumulation in the terrestrial isopod Armadillidium granulatum collected from Ghar El Melh lagoon. We focused on recognizing the effects of trace elements on hepatopancreas functional role. To this end, isopod specimens were exposed for 3 weeks to sediments contaminated with cadmium, copper, zinc, mercury, and nickel. Three concentrations were used in duplicate for each experimental condition. At the end of the experiment, metal body burdens were determined using flame atomic absorption spectrometry. Results of the bioaccumulation factor (BAF) showed that the species A. granulatum was classified as a Cu macroconcentrator (BAF > 2) and a Zn deconcentrator (BAF < 2). Dose dependent morphological and histological changes were observed in the hepatopancreas cells using transmission electron microscopy. The predominant features were: microvillus border disruption, condensation of the cytoplasm with increasing endoplasmic reticulum, mitochondria, lysosomes and granules that accumulated metals in B and S cells. The number of lipid droplets decreased especially after Cd, Zn, Hg, and Ni treatments. This study demonstrated that the terrestrial isopod A. gramulatum could be a good indicator of soil metal contamination., (© 2019 Wiley Periodicals, Inc.)

Chronic wounds unresponsive to existing treatments constitute a serious disease burden. Factors that contribute to the pathogenesis of chronic ulcers include oxidative stress, comorbid microbial infections, and the type of immune system response. Preclinically, and in a case study, a formulation containing a Ceratothoa oestroides olive oil extract promoted wound healing. Patients with chronic venous and pressure ulcers, clinically assessed as being unresponsive to healing agents, were treated for 3 months with an ointment containing the C oestroides extract combined with antibiotic and/or antiseptic agents chosen according to the type of bacterial infection. Treatment evaluation was performed using the Bates-Jensen criteria with +WoundDesk and MOWA cell phone applications. After 3 months of treatment, C oestroides resulted in an average decrease of 36% in the Bates-Jensen score of ulcers ( P < .000), with the decrease being significant from the first month ( P < .007). The combined use of topically applied antibiotics and antiseptics efficiently controlled microbial ulcer infection and facilitated wound healing. In relation to other factors such as initial wound size, chronicity appeared to be an important prognostic factor regarding the extent of wound healing. Future clinical investigations assessing the wound healing efficacy of the C oestroides olive oil extract are warranted.

The sea slater Ligia exotica is believed to have effects of reducing swelling and relieving pain in Chinese folk medicine. However, the scientific foundation of using the sea slater Ligia spp. as an analgesic and anti-inflammatory material remains elusive. In the present study, various organic extracts from sea slater L. exotica were subjected to biological screening employing in vitro and in vivo models, and chemical phenotypes of the biologically active extract were deciphered by integrated gas chromatograph-mass spectrometry (GC-MS) profiling and MS/MS-based molecular networking. The results demonstrated, for the first time, that petroleum ether extract (PE) from L. exotica possessed remarkable anti-inflammatory and analgesic effects. Moreover, intragastric administration of PE at 200 mg/kg produced analgesic effects in both the writhing test and hot plate test. GC-MS analysis revealed that Z-9-hexadecenoic acid and 6-octadecenoic acid dominated in the volatile compositions of PE. Molecular networking (MN) suggested great chemical diversity within L. exotica . In total, 69 known compounds were identified in Ligia extracts by MS/MS spectral matching, and at least 7 analogues from two clusters of nitrogen-containing compounds (MN 3,4 ) were strongly suggested as novel compounds. The molecular families MN 1,3,4 were almost exclusively detected in the biologically active PE and ethyl acetate extract (EE). Importantly, various known compounds identified in MN 1 were reported to possess analgesic and anti-inflammatory effects in the literature, which may contribute to the observed analgesic and anti-inflammatory effects of L. exotica . The present study not only demonstrated the ethnopharmaceutical value of L. exotica for pain-relief in Chinese folk medicine, but also suggested that sea slaters may represent a promising source for discovery of novel analgesic and anti-inflammatory compounds in the near future.

In the last years, the increase of anthropogenic activities has led to a progressive pollution of ecosystems. Oniscidean isopods are a group of terrestrial animals known as excellent bio-indicators and bio-accumulators, because of their capacity to accumulate contaminants and to implement appropriate strategies of detoxification. For this study, we collected Porcellionides pruinosus from two different sites in the Corinth region (Greece), a polluted site and a control site, to assess the heavy metal pollution status. We conducted an inductively coupled plasma mass spectrometry (ICP-MS) analysis for the detection of trace elements in animals' hepatopancreas and soil; we performed also immunohistochemistry for detection of metallothioneins 1 (MT1) and heat shock protein 70 (HSP70s). The results suggest a great capacity of the species in trace elements bioaccumulation and the high degree of pollution of the area near Agioi Theodoroi where the metal pollution index (MPI) is threefolds higher than the control site, due to the presence of significantly higher concentration of arsenic, cadmium, cobalt, chromium, manganese, molybdenum, and lead found in hepatopancreas and soil. The environmental stress is confirmed by the expression of the relative biomarkers of exposure MT1 and HSP70. Our study, with a multimarker approach, consents getting a complete analysis of environmental quality and the effect of pollution on organisms and shows for the first time the heavy metals contamination status in certain Greece area. Furthermore, P. pruinosus proved to be an appropriate indicator of pollution in terrestrial ecosystems., (© 2019 Wiley Periodicals, Inc.)

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.

Among the insulin-family peptides, two additional cysteine residues other than six conserved cysteines are sometimes found in invertebrate insulin-like peptides (ILPs), although the synthetic method for such four disulfide ILPs has not yet been well established. In this study, we synthesized a crustacean insulin-like androgenic gland factor with four disulfides by the regioselective disulfide bond formation reactions using four orthogonal Cys-protecting groups. Its disulfide isomer could be also synthesized by the same method, indicating that the synthetic strategy developed in this study might be useful for the synthesis of other four disulfide ILPs., (© 2018 European Peptide Society and John Wiley & Sons, Ltd.)

Silver nanoparticles (AgNPs) are efficiently converted during the wastewater-treatment process into sparingly soluble Ag sulfides (Ag 2 S). In several countries, sewage sludge is used as a fertilizer in agriculture. The bioavailability of sulfidized Ag to the terrestrial isopod Porcellio scaber was investigated. Sewage sludge containing transformed AgNPs was obtained from a laboratory-scale sewage-treatment plant operated according to Organisation for Economic Co-operation and Development (OECD) guideline 303a. The results of transmission electron microscopy with energy dispersive X-ray of sludge samples suggest that AgNPs were completely transformed to Ag 2 S. Adult isopods were exposed to OECD 207 soil substrate amended with the AgNP spiked sludge for 14 d (uptake phase) followed by an elimination phase in unspiked soil of equal duration. Most of the Ag measured in P. scaber at the end of the uptake phase was found in the hindgut (71%), indicating that only a minor part of the estimated Ag content was actually assimilated by the isopods with 16.3 and 12.7% found in the carcass and hepatopancreas, respectively. As a result of this, the Ag content of the animals dropped following transition to unspiked sludge within 2 d to one-third of the previously measured Ag concentration and remained stable at this level until the end of the elimination period. The present study shows that Ag 2 S in sewage sludge is bioavailable to the terrestrial isopod P. scaber. Environ Toxicol Chem 2018;37:1606-1613. © 2018 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC., (© 2018 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.)

Objective: To assess the antinociceptive and anti-inflammatory properties of the aqueous extract of Armadillidium vulgare (AV)., Methods: The antinociceptive effect of AV (400, 600 and 800 mg/kg) was investigated in mice using the acetic acid-induced writhing, formalin-induced nociceptive, and hot plate tests. Phlogogen-induced paw edema using carrageenan, dextran, or compound 48/80 as phlogogen was used as inflammatory models to evaluate AV's anti-inflammatory effect. Additionally, the bioactive substances glucosamine (GLcN) and taurine in AV were determined using high-performance liquid chromatography., Results: Oral treatment of the mice with AV (600 and 800 mg/kg) significantly reduced the number of writhes in the acetic acid-induced writhing test (P<0.01) but not the hot plate test (P>0.05). All doses tested significantly inhibited paw-withdrawal during the second phase of the formalin-induced nociceptive model (P<0.01). AV demonstrated a strong anti-inflammatory effect in all those inflammatory models (P<0.05)., Conclusions: AV has antinociceptive and anti-inflammatory effects, providing scientific evidence of the efficacy of its traditional use in pain treatment. Furthermore, GLcN and taurine contribute, at least in part, to the anti-inflammatory activity of AV.

In arthropods, the determination of two important parameters of digestive juices, i.e. the total surfactant concentration and the critical micelle concentration (CMC), is challenging due to small sample volumes and low surfactant concentrations. In this work, we report a successful implementation of potentiometric titrations using the surfactant ion-selective electrode (SISE) and the pyrene fluorescence method (PFM) for the determination of the total surfactant concentration and CMC in the digestive juice of terrestrial isopod crustaceans Porcellio scaber. Pooled digestive juice extracts of four (SISE) or two (PFM) animals were used per measurement run. In both cases, digestive juice extracts in 100 μL of deionized water were sufficient for one measurement run. The total surfactant concentration of P. scaber digestive juice was determined to be 9.2 ± 3.5mM and the CMC was approximately 90 μM. Our work presents an important improvement towards easy CMC determination in small volume samples in comparison with the commonly used stalagmometric technique, where much larger sample volumes are usually needed. To date, the total surfactant concentration was not measured in the digestive juices of arthropods other than Homarus vulgaris, Astacus leptodactylus and Cancer pagurus, for which complex separation and analytical techniques were required. Our results obtained by SISE and PFM therefore present the first successful quantification of surfactants and their CMC in small volumes of arthropod digestive juice without prior separation or purification techniques., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Woodlice efficiently sequester copper (Cu) in 'cuprosomes' within hepatopancreatic 'S' cells. Binuclear 'B' cells in the hepatopancreas form iron (Fe) deposits; these cells apparently undergo an apocrine secretory diurnal cycle linked to nocturnal feeding. Synchrotron-based µ-focus X-ray spectroscopy undertaken on thin sections was used to characterize the ligands binding Cu and Fe in S and B cells of Oniscus asellus (Isopoda). Main findings were: (i) morphometry confirmed a diurnal B-cell apocrine cycle; (ii) X-ray fluorescence (XRF) mapping indicated that Cu was co-distributed with sulfur (mainly in S cells), and Fe was co-distributed with phosphate (mainly in B cells); (iii) XRF mapping revealed an intimate morphological relationship between the basal regions of adjacent S and B cells; (iv) molecular modelling and Fourier transform analyses indicated that Cu in the reduced Cu(+) state is mainly coordinated to thiol-rich ligands (Cu-S bond length 2.3 Å) in both cell types, while Fe in the oxidized Fe(3+) state is predominantly oxygen coordinated (estimated Fe-O bond length of approx. 2 Å), with an outer shell of Fe scatterers at approximately 3.05 Å; and (v) no significant differences occur in Cu or Fe speciation at key nodes in the apocrine cycle. Findings imply that S and B cells form integrated unit-pairs; a functional role for secretions from these cellular units in the digestion of recalcitrant dietary components is hypothesized., (© 2016 The Authors.)

In arthropods the cuticle forms an exoskeleton with its physical and chemical properties adapted to functions of distinct skeletal elements. The cuticle of the partes incisivae (PI) in mandibles of terrestrial isopods is a composite of chitin-protein fibrils/fibres and minerals. It consists of an unmineralized tip, a middle region with organic fibrils reinforced mainly with amorphous calcium phosphate and a base region mineralized with amorphous calcium carbonate and calcite. In this study we extend our work on the structure and material properties of the incisive cuticle employing electron backscatter diffraction (EBSD), and investigate calcite orientation patterns in the PI of two terrestrial isopod species from different habitats. We trace small-scale differences in texture sharpness and calcite microstructure, and compare calcite organization and orientation patterns in the PI with those in the tergites of the same isopod species. We observe that in the PI calcite orientation, the degree of crystal alignment, and mode of crystalline domain assemblage is highly varied within short length scales. This contrasts to calcite organization in the tergite cuticle, where calcite has only one specific texture pattern. Such a large range in the variation of calcite organization has not been observed in other carbonate biological hard tissues, such as shells and teeth, where one specific texture and microstructure prevails. Thus, the investigated isopod species are able to control crystallization of the amorphous carbonate precursor in a differential way, most probably related to the function of the individual skeletal element and the animals' behavior., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Objective: To study the chemical constituents of Armadillidium vulgare., Methods: Compounds were isolated and purified by various column chromatography, spectroscopic methods were used to identify their structures., Results: Seven compounds were isolated from Armadillidium vulgare and identified as vulgarine A (1), o-hydroxylbenzoic acid (2), dipyrrolopiperazine-2,5-dione (3), 4(1-H)-quinolone (4), adenine (5), n-acetyltyramine (6) and 4-methyl-5-thiazoleethanol (7)., Conclusion: Compound 1 is a new alkaloid, compounds 3-7 are all nitrogen containing substances, which are isolated from Armadillidae family for the first time.

In terrestrial isopods the mandibles consist of a corpus carrying strong muscle tissue, and a pars incisiva (PI) that cuts dry leaves into smaller ingestible pieces. We studied the cuticle of the PI of Porcellio scaber in order to understand region-dependent differences in its ultrastructure, composition, and the resulting mechanical properties, employing several microscopic and analytical techniques as well as nanoindentation experiments. The cuticle of the incisive tip is not mineralized and consists of an unusually thick epicuticle containing thin fibrils, two subjacent cuticular layers, and a central core containing fibrils of different orientation, either longitudinal or circumferential. A thick epicuticle of the middle region just behind the tip projects long epicuticular extensions into the subjacent endocuticle, likely to prevent delamination. A distinct exocuticular layer is lacking in the middle region. Most chitin-protein fibrils within the endocuticle are oriented in parallel pointing towards the tip. Surprisingly, the middle region is mineralized by amorphous calcium phosphate (ACP) only. Near the base, ACP is successively replaced by amorphous calcium carbonate and calcite is restricted to a distal layer in the base. At the transition between middle and base, the epicuticle forms a hybrid material containing fibrils of the exocuticle. Nanoindentation experiments reveal an increase of the stiffness and hardness from the tip towards the base and significantly higher values on transversal in comparison to longitudinal planes. The results suggest that ultrastructure and composition are adapted for conveying high forces from a rather thin cutting edge to the stable base of the PI., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The cuticle of the cornea in Crustacea is an interesting example of a composite material compromising between two distinct functions. As part of the dioptric apparatus of the ommatidia within the complex eye it forms transparent micro-lenses that should as well maintain the mechanical stability of the head capsule. We analyzed the ultrastructure and composition of the isopod cornea cuticle of the terrestrial species Ligia oceanica and the marine Sphaeroma serratum. We used a variety of tissue preparation methods, electron microscopic techniques as well as electron microprobe analysis and Raman spectroscopic imaging. The results reveal various structural adaptations that likely increase light transmission. These are an increase in the thickness of the epicuticle, a reduction of the thickness of the outer layer of calcite, a spatial restriction of pore canals to interommatidial regions, and, for S. serratum only, an increase in calcite crystal size. In both species protein-chitin fibrils within the proximal exocuticle form a peculiar reticular structure that does not occur within the cuticle of the head capsule. In L. oceanica differential mineralization results in a spherically shaped interface between mineralized and unmineralized endocuticle, likely an adaptation to increase the refractive power of the cornea maintaining the mechanical stability of the cuticle between the ommatidia. The results show that the habitat and differences in the general structure of the animal's cuticle affect the way in which the cornea is adapted to its optical function., (Copyright © 2014 Elsevier Inc. All rights reserved.)

When planning rodent eradications, that normally involve the use of the anticoagulant poison brodifacoum, it is imperative to minimise impacts on other "non-target" species that dwell alongside the targeted rodents and may indeed be the intended beneficiaries of the eradication. Such impacts can arise either from primary poisoning when the non-target species ingest bait pellets containing toxicant or by secondary poisoning when the non-target species eats prey that has itself eaten brodifacoum. Cockroaches and woodlice, likely to scavenge bait pellets, are widely distributed on tropical and sub-tropical islands where they are eaten by ground-dwelling birds. Combining work on Henderson Island, South Pacific, site of a recent rat eradication project, and UK laboratory experiments, our study first measured brodifacoum concentrations in cockroaches given temporary ad lib access to poison bait pellets, approximately mimicking the aftermath of bait distribution for a rodent eradication. In two separate experiments using different species/exposure times, the mean brodifacoum concentrations among cockroaches immediately after bait exposure was 262±s.e. 131 and 477±168µgkg(-1) wet weight. Values decreased quickly in the following 2 weeks, and then continued to decline at a slower rate over the following 4 weeks in the more prolonged laboratory experiment. A supplementary experiment with woodlice recorded a similar brodifacoum concentration in the animals at the end of the exposure period, 223±66µgkg(-1), and a similar time course for the post-exposure decline. In the context of rails (Rallidae), a group of birds known to be particularly susceptible to primary brodifacoum poisoning, these results suggested that, in terms of acute exposure, individual birds would need to eat a minimum of their own body weight (and more commonly 2-5 times that) of live cockroaches before facing a 50% risk of death. Therefore, we conclude that in eradication scenarios, acute secondary poisoning is of lower concern for these birds than primary poisoning., (© 2013 Elsevier Inc. All rights reserved.)

The major environmental impact of landfills is emission of pollutants via the leachate and gas pathways. The hepatopancreas of the terrestrial isopod Armadillidium vulgare (Isopoda, Crustacea, Latreille 1804) plays an important role in the bioaccumulation of contaminants, such as heavy metals. To evaluate the effects of landfill leachate treatment, 2 different approaches were applied: 1) the detection of accumulation of trace elements (As, Cd, Cr, Cu, Sb, Zn, Pb, Ni, V) in hepatopancreatic cells, and 2) the evaluation of biological effect of contaminants on fresh hepatopancreatic cells by flow-cytometric analyses. The presence of 2 different cell types (herein referred to as "small" [S] cells and "big" [B] cells, in agreement with the literature based on morphological examinations) was detected for the first time by flow cytometry, which also highlighted their different response to stress stimuli. In particular, B cells appeared more sensitive to landfill leachate treatment, being more damaged in the short term, while S cells seemed more adaptive. Furthermore, S cells could represent a pool from which they are able to differentiate into B cells. These findings were also confirmed by principal component analyses, underlining that S SYBR Green I bright cells correlate with specific chemicals (Ca, Cu, Co), confirming their resistance to stress stimuli, and suggesting that the decrease of specific cell types may prime other elements to replace them in a homeostasis-preservation framework., (© 2013 SETAC.)

The cuticle of terrestrial isopods is an interesting model for the study of structure-function relationships in biological composite materials. Its organic matrix has a hierarchically organised structure, and type and phase of the mineral compound can vary. The cuticle forms functionally diverse skeletal elements whose properties are adapted to their specific functions. In order to better understand the relation between structure, composition and function of isopod cuticle, we studied the structure and composition of the joint head that is part of the pereiopod's basis. It consists of a central region, whose shape fits well into the joint socket, and an edge region that is connected to the soft arthrodial membrane and protects the central region from mechanical load. The cuticle architecture of the joint head has local variations in structure and composition. In the central region the cuticle is similar to the previously published tergite cuticle. High concentrations of amorphous calcium phosphate are located in the endocuticle suggesting a coexistence with amorphous calcium carbonate. The edge region has an unexpected organisation characterised by thickening of the epi- and exocuticle and an unusual unidirectional orientation of chitin-protein fibrils within the endocuticle. The concentrations of phosphate are considerably higher than in the central region. The overall differentiation in the cuticular architecture of the edge in comparison to the central region reflects the adaptation to mechanical strains the cuticle has to sustain during contraction of extensor muscles, and to the structural and compositional transition from the edge to the connecting arthrodial membrane., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Mercury (Hg) is a trace element with high toxicological impact on potential receptors, including human beings. Global Hg emissions are predicted to increase significantly during the next 40 years. After emission, the metal is transported by air currents and precipitations, leading to increasing depositions even in areas far from emission sources. In the terrestrial environment, Hg is subjected to redistribution and transformation into different inorganic and metal-organic species that are taken up by vegetation and soil organisms. In the present study, the woodlouse (Porcellio scaber) was used as a biological indicator of total Hg pollution in the city of Dornbirn (province of Vorarlberg), Austria. Woodlice were collected from 30 sampling points scattered over the city area, 25 of them situated within a rectangular transect crossing the city area from west-northwest to east-southeast, starting near the Rheintal motorway and ending at the slopes of the Bregenzer Wald hills. In addition to woodlice, soil substrate samples were collected at nine of the selected sampling points. Total Hg concentrations were measured in isopod tissues and soil substrate samples by means of an Hg analyzer. Total Hg concentrations in isopod tissues were significantly correlated with Hg soil contents (P < 0.05). Moreover, a gradient of increasing Hg concentrations was observed in isopod samples along the transect across Dornbirn, with the lowest concentrations detected in woodlouse samples near the Rheintal motorway and the highest levels toward the ascending slopes of the Bregenzer Wald hills. This gradient of increasing Hg concentrations across the city matches a concomitant increase in wet precipitations along the same direction, indicating that deposition by wet precipitation may be an important source for Hg contamination in the city of Dornbirn. Overall, the degree of Hg contamination across the study area can be regarded as rather low, i.e., comparable with concentrations observed in other, unpolluted terrestrial habitats. It is concluded that bioindication by total Hg analysis in woodlice can be applied to distinguish between different levels and sources of contamination in urban areas.

Objective: To identify consistency of the composition and efficacy of Porcellio scaber and Armadillidium vulgare and provide reference for the safe and scientific use of the medicinal material., Methods: Identified the Porcellio scaber and the Armadillidium vulgare with the method of characteristics; Compared the two chemical composition by thin layer method; Determined the extract by water and ethanol using the pharmacopoeia extract method., Results: Porcellio scaber and Armadillidium vulgare were two varieties, they had similar chemical composition, the water and ethanol extract were the same., Conclusion: The compostion and efficary of Porcellio scaber and Armadillidium vulgare are the same, and these two varieties can be mixed or used alone.

The main mineral components of the isopod cuticle consists of crystalline magnesium calcite and amorphous calcium carbonate. During moulting isopods moult first the posterior and then the anterior half of the body. In terrestrial species calcium carbonate is subject to resorption, storage and recycling in order to retain significant fractions of the mineral during the moulting cycle. We used synchrotron X-ray powder diffraction, elemental analysis and Raman spectroscopy to quantify the ACC/calcite ratio, the mineral phase distribution and the composition within the anterior and posterior tergite cuticle during eight different stages of the moulting cycle of Porcellio scaber. The results show that most of the amorphous calcium carbonate (ACC) is resorbed from the cuticle, whereas calcite remains in the old cuticle and is shed during moulting. During premoult resorption of ACC from the posterior cuticle is accompanied by an increase within the anterior tergites, and mineralization of the new posterior cuticle by resorption of mineral from the anterior cuticle. This suggests that one reason for using ACC in cuticle mineralization is to facilitate resorption and recycling of cuticular calcium carbonate. Furthermore we show that ACC precedes the formation of calcite in distal layers of the tergite cuticle., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The crustacean cuticle is a hierarchically organised material composed of an organic matrix and mineral. It is subdivided into skeletal elements whose physical properties are adapted to their function and the eco-physiological strains of the animal. Using a variety of ultrastructural and analytical techniques we studied the organisation of the tergite cuticle of the sand burrowing beach isopod Tylos europaeus. The surface of the tergites bear epicuticular scales, sensilla and micro-tubercles. A distal layer of the exocuticle is characterised by a low density of organic fibres and the presence of magnesium-calcite. Surprisingly, the mineral forms regions containing polyhedral structures alternating with smooth areas. Between sub-domains within the distal exocuticle calcite varies in its crystallographic orientation. Proximal layers of the exocuticle and the endocuticle are devoid of calcite and the mineral occurs in the form of amorphous calcium carbonate (ACC). Using thin sections of mineralised cuticle we describe for the first time that ACC forms tubes around single protein-chitin fibrils., (Copyright © 2011 Elsevier Inc. All rights reserved.)

The RP HPLC and LC/MS QTOF analyses of the methanolic CNS extract from isopod crustacean the woodlouse, Porcellio scaber revealed a presence of the red pigment-concentrating hormone (Panbo-RPCH) in this species. It has been shown that this neuropeptide plays a role in mobilization of energy stores: topical treatments of P. scaber individuals by Panbo-RPCH in a concentration 20 pmol/microl increased the level of glucose in haemolymph about 4 times, while the level of trehalose was only doubled. The results demonstrated that glucose was the main carbohydrate mobilized by the Panbo-RPCH treatment: glucose was responsible for about 97% of total carbohydrate increasing. Despite the demonstration of hyperglycaemic activity of Panbo-RPCH, no stimulatory effect of this hormone on the locomotory activity of P. scaber was observed. The present study is the first discovery of an occurrence of Panbo-RPCH and its hyperglycaemic activity in the representative of the isopod crustaceans. The relationship of the function of Panbo-RPCH in P. scaber to the role of this neuropeptide and adipokinetic hormones in insects is discussed., ((c) 2009 Elsevier Inc. All rights reserved.)

We used Yellow-legged Gull (YLG) chicks to monitor trace elements in Tunisian areas subject to different pollution stresses: urban contamination (Chikly), industrial pollution (Thyna) and an unpolluted area (Kneis). We measured trace element concentrations (Hg, Se and Pb) in chick feathers. We also assessed their feeding ecology by analyzing both regurgitates and stable isotopes (SIA) in chick feathers and in their prey, to determine the main entry route of pollutants. SIA revealed that YLG feed mainly on aquatic resources from the Lake of Tunis (Chikly colony) and the Gulf of Gabès (Thyna and Kneis colonies). Moreover, the enriched delta(15)N found in feathers from Chikly are attributed to the eutrophication of the Lake of Tunis. Hg and Se were higher in Kneis and Thyna colonies, in agreement with the higher consumption of marine resources and the greater availability of these elements resulting from the impact of the industrial activity in the area. Pb concentrations were higher in Chikly, related to the heavier traffic around the Lake of Tunis and the use of leaded gasoline., (Copyright 2009 Elsevier Ltd. All rights reserved.)

The present study investigated the contribution of environmental factors to the accumulation of As, Cd, Cu, Pb and Zn in earthworms, beetles and woodlice, and framed within an exposure assessment of the European hedgehog. Soil and invertebrate samples were collected in three distinct habitat types. Results showed habitat-specific differences in soil and invertebrate metal concentrations and bioaccumulation factors when normalized to soil metal concentration. Further multiple regression analysis showed residual variability (habitat differences) in bioaccumulation that could not be fully explained by differences in soil metal contamination, pH or organic carbon (OC). Therefore, the study demonstrated that in bioaccumulation studies involving terrestrial invertebrates or in risk assessment of metals, it is not sufficient to differentiate habitat types on general soil characteristics such as pH and/or OC alone. Furthermore, simple generic soil risk assessments for Cd and Cu showed that risk characterization was more accurate when performed in a habitat-specific way.

Leaching using EDTA applied to a Pb, Zn and Cd polluted soil significantly reduced soil metal concentrations and the pool of metals in labile soil fractions. Metal mobility (Toxicity Characteristic Leaching Procedure), phytoavailability (diethylenetriaminepentaacetic acid extraction) and human oral-bioavailability (Physiologically Based Extraction Test) were reduced by 85-92%, 68-91% and 88-95%, respectively. The metal accumulation capacity of the terrestrial isopod Porcellio scaber (Crustacea) was used as in vivo assay of metal bioavailability, before and after soil remediation. After feeding on metal contaminated soil for two weeks, P. scaber accumulated Pb, Zn and Cd in a concentration dependent manner. The amounts of accumulated metals were, however, higher than expected on the basis of extraction (in vitro) tests. The combined results of chemical extractions and the in vivo test with P. scaber provide a more relevant picture of the availability stripping of metals after soil remediation.

The concentrations of Ag, Cd, Co, Cr, Cu, Fe, Ni, Pb, Mn and Zn in Saduria entomon and adjacent bottom sediments from the southern Baltic were determined by FAAS. In order to estimate the strength of correlations between accumulated elements in these crustaceans and surficial sediment, bioaccumulation factors (BAFs) were calculated. The results of factor analysis (FA) and the Kruskal-Wallis analysis of variance (ANOVA) clearly indicate geographical differences between the concentrations of these elements. Cd, Co, Fe, Ni, Pb and Zn levels were higher in S. entomon from the Gulf of Gdańsk, whereas Cr and Mn levels were higher in the crustaceans inhabiting open Baltic waters. The concentrations of Ag and Cu were comparable in both regions. There was a tendency for metal concentrations to distinguish organisms inhabiting the muddy bottom from those living in sandy sediments. The granulometric composition of the sediment appears to influence trace metal bioavailability. The results show that S. entomon could be a valuable sentinel organism for biomonitoring heavy metal contamination in the southern Baltic.

To understand why Arthrobotrys oligospora and other nematode-trapping fungi are common and sometimes abundant in the coastal grassland soils of the Bodega Marine Reserve (BMR, Sonoma County, CA), we examined how resident trapping fungi responded to the addition of eight organic substrates (lupine leaves, grass leaves, dead isopods, dead moth larvae, isopod faeces, deer faeces, shrimp shells, and powdered chitin). We were especially interested in the effects of dead isopods because isopods are abundant at BMR and because previous studies had documented strong responses of A. oligospora to other arthropods (dead moth larvae). Soil from BMR was packed into vials (40 g dry mass equivalent per vial with water potential at -230 kPa and bulk density at 0.9 gcm(-3)), and one substrate or no substrate was added to the soil surface. After 30 d at 20 degrees C, trapping fungi were quantified by dilution plating and most probable number procedures. The response of A. oligospora was inversely related to substrate carbon:nitrogen (C:N) ratio: substrates with low C:N ratios (dead isopods, lupine leaves, dead moth larvae) usually caused large increases in A. oligospora whereas those with higher C:N ratios (isopod faeces, deer faeces, grass leaves) did not. An exception was chitin powder, which had a low C:N ratio, but which did not cause A. oligospora to proliferate. Responses of A. oligospora were directly related to the quantity of nitrogen added with each substrate, and those substrates that caused large increases in resident nematodes usually caused large increases in A. oligospora. Other trapping fungi did not respond as strongly as A. oligospora.

Anecdotal observations of the Dougherty plain cave crayfish (Cambarus cryptodytes), the Georgia blind cave salamander (Haideotriton wallacei), and albinistic isopods (Caecidotea sp.) at great depths below the land surface and distant from river corridors suggest that obligate aquifer-dwelling (troglobitic) organisms are widely distributed throughout the limestone Upper Floridan aquifer (UFA). One mechanism by which subterranean life can proliferate in an environment void of plant productivity is through a microbial food web that includes chemosynthesis. We examined this possibility in the UFA by measuring the isotopic composition ((13)C, (14)C, and (15)N) of tissues from troglobitic macrofauna. Organisms that were captured by cave divers entering into spring conduits had delta(13)C values that suggested plant matter as a primary food resource (cave crayfish, -24.6 +/- 2.7 per thousand, n = 9). In contrast, delta(13)C values were significantly depleted in organisms retrieved from wells drilled into areas of the UFA remote from spring and sinkhole conduits (cave crayfish -34.7 +/- 9.8 per thousand, n = 10). Depleted (13)C values in crayfish were correlated with radiocarbon (Delta(14)C) depletion relative to modern values. The results suggest that methane-based microbial chemosynthetic pathways support organisms living in the remote interior of the aquifer, at least in part.

The aim of the present study is to evaluate the seasonal fluctuation of heavy metals in the isopod Porcellio laevis at four uncontaminated subtropical locations. This study was carried out at four different field sites in Assiut, Egypt. The concentrations of cadmium, lead, copper, and zinc in animal, soil, and litter (mug/g dry weight) were monthly recorded during the period from June 2002 till May 2003. There was little difference in metal accumulation trends between the sites. In general, the isopod showed significant increased Pb and Zn concentration during summer and spring months, whereas this was not the case for Cd and Cu. The bioaccumulation (BAF) and bioconcentration factors (BCF) of the metals revealed marked seasonal changes throughout the year. Generally, BAF of metals were higher during summer and spring, and BCF were higher during summer and autumn. Comparing the metal accumulation with climatic fluctuations (measured) it was speculated that temperature was the main factor causing seasonal fluctuations of the internal metal concentration in the isopod.

The present study was conducted to find potential terrestrial biomonitors for heavy metals in indoor air in an urban environment. TSP, PM(10), and PM(2.5) were collected in three retirement facilities in the urban area of Vienna. In addition, particulate matter and soil, vegetation, and isopods (Porcellio scaber L.) were collected in the adjacent garden areas. Aerosols were sampled with a low-volume air sampler. The sampled materials were wet ashed and total lead and cadmium contents were determined. Water-soluble heavy metal concentrations were measured in aqueous extracts from air exposed filters, soil, and vegetation. Lead and cadmium were analyzed by graphite furnace AAS. Lead contents in the vegetation were inferred from water-soluble lead in soils. Lead in isopods generally reflected the contents in vegetation. Cadmium in plants probably derived from soil solutions as well as from atmospheric input. Isopods reflected the total cadmium contents in soils. Particulate matter was dominated by PM(2.5), both with respect to mass concentrations and to heavy metal contents. The indoor aerosol was found to be influenced by human activity, indoor sources, and outdoor particles. Relationships between indoor airborne heavy metals and the contents in vegetation (lead and cadmium: positive) and isopods (lead: negative) were identified to have the potential for biomonitoring indoor air quality.

Before the molt terrestrial isopods resorb calcium from the posterior cuticle and store it in large deposits within the first four anterior sternites. In Porcellio scaber the deposits consist of three structurally distinct layers consisting of amorphous CaCO3 (ACC) and an organic matrix that consists of concentric and radial elements. It is thought that the organic matrix plays a role in the structural organization of deposits and in the stabilization of ACC, which is unstable in vitro. In this paper, we present a thorough analysis of the ultrastructure of the organic matrix in the CaCO3 deposits using high-resolution field-emission scanning electron microscopy. The spherules and the homogeneous layer contain an elaborate organic matrix with similar structural organization consisting of concentric reticules and radial strands. The decalcification experiments reveal an inhomogeneous solubility of ACC within the spherules probably caused by variations in the stabilizing properties of matrix components. The transition between the three layers can be explained by changes in the number of spherule nucleation sites.

Cd, Cr, Cu and Ni were determined in two different species of woodlice: Porcellio scaber and Porcellio dilatatus. Both P. scaber and P. dilatatus were cultivated under standardized conditions in a climatic chamber. Moreover, skins of the cultivated animals were collected and analysed separately to examine whether moulting is a way of detoxification from these elements. After drying and grinding both animal and skin samples, they were pooled to obtain enough sample material for each species. The pooled samples were digested in pure concentrated nitric acid using microwave-assisted high pressure digestion and, finally, analysed by ICP-OES. Special emphasis was given to quality control. To match the matrix of the samples very closely, the reference materials Dorm-2, Dogfish Muscle (Squalus acanthias) and SRM 1577b Bovine Liver were used to evaluate the whole analytical process including sample digestion. Analyses of the elements in the reference materials were carried out using three different wavelengths for each element simultaneously to check for spectral interferences and to select the wavelengths which were best suited for the analyses. Concentrations of Cd, Cr, Cu and Ni in woodlice and their skins indicated that moulting is a possible means of detoxification in the case of Cr and Ni.

We report the isolation and the characterization of a novel antibacterial peptide from hemocytes of the woodlouse Armadillidium vulgare, naturally infected or uninfected by Wolbachia, an intracellular Gram-negative bacterium. This molecule displays antibacterial activity against Gram-positive bacteria despite its composition which classes it into the glycine-rich antibacterial peptide family, usually directed against fungi and Gram-negative bacteria. The complete sequence was determined by a combination of Edman degradation, mass spectrometry and cDNA cloning using a hemocyte library. The mature peptide (53 residues) has a 5259 Da molecular mass and is post-translationally modified by a C-terminal amidation. This peptide is characterized by a high level of glycine (47%) and a fivefold repeated motif GGGFH(R/S). As no evident sequence homology to other hitherto described antibacterial peptides has been found out, this antibacterial peptide was named armadillidin. Armadillidin is constitutively expressed in hemocytes and appears to be specific of A. vulgare.

Column (CC), thin-layer (TLC), high-performance liquid (HPLC) and ion-exchange chromatography (IEC), were used to investigate corotenoid and carotenoprotein complexes in Asellus aquaticus specimens from the Narew river. The following carotenoids were found: alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein, zeaxanthin, diadinoxanthin, mutatoxanthin, crustaxanthin, echinenone, hydroxyechinenone, phoenicoxanthin, canthaxanthin and astaxanthin. Astaxanthin (37.5%), canthaxanthin (21.4%) and phoenicoxanthin (12.3%) were found in the largest amounts. The total carotenoid content was 13.824 microg g(-1) of dry mass. Carotenoprotein complexes containing astaxanthin as the prosthetic group were purified from Asellus aquaticus. The carotenoprotein complexes belonged to the crustacyanins group as alpha- and gamma-crustacyanin. The protein forming the alpha-crustacyanin contained large amounts of such amino-acids as asparic acid, glutamic acid and leucine, whereas the protein of the gamma-crustacyanin contained primarily glutamic acid, glycine and lysine.

Floodplains of the European rivers Rhine and Meuse are heavily polluted. We investigated the risk of heavy metal pollution (Cd, Cu, Pb, Zn) for detritivores living in a floodplain area, the Biesbosch, the Netherlands, affected by these rivers. Total soil, pore water and 0.01 M CaCl(2) extractable concentrations and concentrations in plant leaves, earthworms, isopods and millipedes were measured in two sites and compared with literature data to assess possible risks. Based on total metal concentrations in soil, serious effects on detritivores were expected. However, 0.01 M CaCl(2) extractable, pore water and plant leaf concentrations were similar to metal concentrations found in unpolluted areas. Concentrations of Cu and Cd in earthworms and Cu in millipedes were higher in the Biesbosch than in animals from reference areas. All other measured concentrations of heavy metals in earthworms, isopods and millipedes were similar to the ones found in reference areas. Despite high total soil concentrations, effects of Zn, Cu, Pb and Cd pollution on isopods are therefore not expected, while millipedes may only be affected by Cu. Since Cu and Cd levels in earthworms were increased compared to animals in unpolluted soils, this faunal group seems to be most at risk. Given the engineering role of earthworms in ecosystems, effects on the ecological functioning of floodplain soils therefore cannot be excluded.

It appears from grafting experiments that the androgenic hormone (AH) of terrestrial isopods has a narrow species-specificity [J. Crust. Biol. 19 (1999) 684], even if AH of different species shared common epitopes [Gen. Comp. Endocrinol. 125 (2002) 218]. To date only the glycosylated AH of the woodlouse Armadillidium vulgare has been deciphered by us [Ann. New York Acad. Sci. 839 (1998) 111; Eur. J. Biochem. 262 (1999) 727] and [Biochem. Biophys. Res. Commun. 264 (1999) 419] have confirmed the primary structure of this protein. We reported in the present paper the characterization of the AH in another species Porcellio scaber by a combination of microsequencing, mass spectrometry, and molecular cloning. We found only one glycoform which consisted of two peptide chains, A and B, of 31 and 44 amino acids, respectively, with A chain carrying on Asn18 a N-glycosylated moiety, size of which has been determined by MALDI-MS measurements. The expected structure of the glycosylation was proposed. The deduced amino acid sequence of the AH precursor was mainly identical to the one obtained independently by another group [Zool. Sci. 20 (2003) 75]. We also showed that AH gene is exclusively expressed in androgenic glands. Sequence comparison with A. vulgare and P. scaber (population of Japan) AH was discussed.

When exposed to methylmercury in the laboratory, grass shrimp, Palaemonetes pugio, parasitized by the isopod Probopyrus pandalicola, accumulated lower concentrations of mercury than their unparasitized counterparts. The parasitic isopod accumulated far less mercury than the grass shrimp. When exposed to mercury in a contaminated field site, mummichogs, Fundulus heteroclitus, parasitized with the nematode Eustrongylides, similarly accumulated lower concentrations of mercury than unparasitized fish, and the parasite similarly accumulated less than the host. The lower uptake by the parasites compared to their hosts is counter to the general view of biomagnification of methylmercury, since parasites are a trophic level above their hosts. The mechanism whereby parasitized animals accumulate less toxicant than unparasitized ones is unknown, but may be partially due to lower metabolic rate.

It is thought that a plasma membrane Ca(2+)-transport ATPase (PMCA) and a Na(+)/Ca(2+)-exchange (NCE) mechanism are involved in epithelial Ca(2+) transport (ECT) in a variety of crustacean epithelia. The sternal epithelium of the terrestrial isopod Porcellio scaber was used as a model for the analysis of Ca(2+)-extrusion mechanisms in the hypodermal epithelium. Using RT-PCR, we amplified a cDNA fragment of 1173 bp that encodes a protein sequence possessing 72% identity to the PMCA from Drosophila melanogaster and a cDNA fragment of 791 bp encoding a protein sequence with 50% identity to the NCE from Loligo opalescens. Semiquantitative RT-PCR revealed that the expression of both mRNAs increases from the non-Ca(2+)-transporting condition to the stages of CaCO(3) deposit formation and degradation. During Ca(2+)-transporting stages, the expression of PMCA and NCE was larger in the anterior sternal epithelium (ASE) than in the posterior sternal epithelium (PSE). The results demonstrate for the first time the expression of a PMCA and a NCE in the hypodermal epithelium of a crustacean and indicate a contribution of these transport mechanisms in ECT.