(E)-7-phenyl-2-heptene-4,6-diyn-1-ol from Bidens pilosa as a repellent against isopods.

The development of repellents as alternatives to insecticides has expanded in recent years. However, their use in isopod pest control is limited. To develop an isopod repellent, a plant extract library from wild plants native to the Kochi Prefecture was screened for repellent activity against pillbugs, and 82 samples (87%) exhibited repellent activity. Among them, (E)-7-phenyl-2-heptene-4,6-diyn-1-ol was isolated and identified as a repellent from the root of Bidens pilosa. It had a half-maximal effective concentration of 0.20 µm, with a strong repellency. A study of the structure-activity relationship to (E)-7-phenyl-2-heptene-4,6-diyn-1-ol revealed that the presence of a hydroxyl group and an aromatic at both ends of the length of the seven-carbon chain is important for the expression of repellency. These results can potentially lead to a new repellent of phenylalkyl alcohol.

Effects of combined treatment of cadmium and oxytetracycline on the terrestrial isopod Porcellio leavis / Efeitos do tratamento combinado de cádmio e oxitetraciclina no isópode terrestre Porcellio laevis

The influence of pharmaceutical residues and heavy metals on living organisms has received global attention. The present study assessed the interactive effect of antibiotic residues and heavy metals in soil, as contaminated food with cadmium (Cd) and oxytetracycline (OTC) on the isopod Porcellio leavis. It was fed on fresh plant leaves contaminated with different concentrations of cadmium, Cd+OTC1000 ppm, Cd+OTC2000 ppm and Cd+OTC3000 ppm for 4 weeks. The changes in the feeding patterns, protein, lipid peroxidation (LPO), catalase activity (CAT), and total free amino acids (TFAA) were recorded. There were significant differences in the obtained results where Cd reduced the egestion ratio (ER) however, OTC enhanced this ratio. Biochemical analysis illustrated that combination between OTC and Cd inhibits the toxic effects of Cd at low concentration (1000 ppm), while at high concentration (3000 ppm) raise the toxicity. Detailed studies are required for further understanding of the interaction between OTC and heavy metals, and also its impact on soil animals and for improving soil risk evaluation.

A influência de resíduos farmacêuticos e metais pesados em organismos vivos tem recebido atenção global. O presente estudo avaliou o efeito de resíduos de antibióticos e metais pesados no solo, como alimentos contaminados com cádmio (Cd) e oxitetraciclina (OTC), sobre o isópode Porcellio laevis, o qual foi alimentado com folhas frescas de plantas contaminadas com diferentes concentrações de cádmio, Cd + OTC1000 ppm, Cd + OTC2000 ppm e Cd + OTC3000 ppm, por quatro semanas. As mudanças nos padrões de alimentação, proteína, peroxidação lipídica (LPO), atividade da catalase (CAT) e aminoácidos livres totais (TFAA) foram registradas. Houve diferenças significativas nos resultados obtidos, em que o Cd reduziu a taxa de excreção (ER), no entanto o OTC aumentou essa proporção. A análise bioquímica mostrou que a combinação entre OTC e Cd inibe os efeitos tóxicos do Cd em baixa concentração (1.000 ppm), enquanto, em alta concentração (3.000 ppm), aumenta a toxicidade. Estudos detalhados são necessários para uma maior compreensão da interação entre OTC e metais pesados, e seu impacto sobre os animais do solo, bem como para melhorar a avaliação de risco do solo.

Effects of combined treatment of cadmium and oxytetracycline on the terrestrial isopod Porcellio leavis.

The influence of pharmaceutical residues and heavy metals on living organisms has received global attention. The present study assessed the interactive effect of antibiotic residues and heavy metals in soil, as contaminated food with cadmium (Cd) and oxytetracycline (OTC) on the isopod Porcellio leavis. It was fed on fresh plant leaves contaminated with different concentrations of cadmium, Cd+OTC1000 ppm, Cd+OTC2000 ppm and Cd+OTC3000 ppm for 4 weeks. The changes in the feeding patterns, protein, lipid peroxidation (LPO), catalase activity (CAT), and total free amino acids (TFAA) were recorded. There were significant differences in the obtained results where Cd reduced the egestion ratio (ER) however, OTC enhanced this ratio. Biochemical analysis illustrated that combination between OTC and Cd inhibits the toxic effects of Cd at low concentration (1000 ppm), while at high concentration (3000 ppm) raise the toxicity. Detailed studies are required for further understanding of the interaction between OTC and heavy metals, and also its impact on soil animals and for improving soil risk evaluation.

Soil washing with biodegradable chelating agents and EDTA: Technological feasibility, remediation efficiency and environmental sustainability.

In washing soils contaminated with toxic metals, the replacement of recalcitrant EDTA with biodegradable chelators has gained high expectations. Herein we investigated the feasibility of using EDTA and biodegradable GLDA, EDDS and IDS under conditions pertinent to operational remediation technology, in a pilot-scale experiment. GLDA and IDS did not precipitate from process solutions, which lessened their recyclability. In other process parameters, chelator supplement, Na-saturation of process solutions and processing time, EDTA outperformed biodegradable chelators. Treatment with EDTA was also the most effective in total Pb and Zn removal and least impacted soil properties. GLDA was slightly better in Cd removal. EDDS and IDS were inefficient. All chelators effectively removed easily-available Pb, Zn and Cd from the exchangeable soil fraction. EDTA was the most efficient chelator in reducing the bioaccessibility of Pb and GLDA in reducing the bioaccessibility of Cd from simulated human gastrointestinal tract. Treatment with GLDA had an edge in reducing plant bioaccessibility of toxic metals, but induced worrying leachability of Pb. This was 8.3-times higher than with the process with EDTA and 3.4-times higher than in original soil. In general, our results demonstrate the advantage of EDTA over tested biodegradable chelators in process and remediation efficiency and environmental safety.

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.

Integrated Gas Chromatograph-Mass Spectrometry (GC/MS) and MS/MS-Based Molecular Networking Reveals the Analgesic and Anti-Inflammatory Phenotypes of the Sea Slater Ligia exotica.

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 (MN3,4) were strongly suggested as novel compounds. The molecular families MN1,3,4 were almost exclusively detected in the biologically active PE and ethyl acetate extract (EE). Importantly, various known compounds identified in MN1 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.

The effect of soil pollution with petroleum-derived substances on Porcellio scaber Latr. (Crustacea, Isopoda).

Presented research aimed at investigating the effect of short-term contact with petroleum-derived substances (PDSs) on life parameters of Porcellio scaber Latr. (Isopoda) and accumulation of polycyclic aromatic hydrocarbons (PAHs) in its body. The influence of presence of P. scaber on the total petroleum hydrocarbons (TPH) content in soil was also determined. The following objects were established: control-unpolluted soil; soil polluted with petrol; soil polluted with diesel fuel and soil polluted with used engine oil. Every pollutant was used in the amounts equal to 6000 mg of fuel/kg d.m. of soil 15 months earlier. In the laboratory, survival and body mass change of P. scaber reared in investigated soils were observed. The delivered food was not contaminated with PDSs. P. scaber reveals a considerable resistance in a short (4 weeks) contact with PDSs, evidenced as high survivability (from 68% on the soil polluted with engine oil to 77% on the soil polluted with diesel fuel) and undisturbed increase in body mass (on the level similar to control). It indicates the potential usefulness of this animal as a monitoring organism. No positive correlation was observed between TPH depletion in the soils contaminated with PDSs and P. scaber presence during 4 weeks of the experiment. PAH level in P. scaber bodies was generally very low (with the highest level of anthracene 0.40 µg/g of wet mass-after 4 weeks of rearing on the diesel fuel-contaminated soil), which may confirm the thesis about considerable abilities of isopods for biotransformation of these pollutants and low susceptibility to these xenobiotic penetration through integuments. However, a tendency for accumulation for phenanthrene and anthracene in conditions of soil polluted with diesel fuel was observed respectively 0.07 and 0.21 µg/g of wet mass for phenanthrene and 0.22 and 0.40 µg/g of wet mass for anthracene, observed successively in the 2nd and 4th week of rearing.

A new species of Pseudione Kossmann, 1881 (Isopoda, Cymothoida, Bopyridae) parasitizing the lobster Nephropsis aculeata Smith, 1881 (Decapoda, Astacidea, Nephropidae) in the Southwestern Atlantic.

A new species of Pseudione Kossmann, 1881 is described from two parasitized specimens of the clawed lobster Nephropsis aculeata Smith, 1881, collected from Brazilian waters in the southwestern Atlantic. It is the second species of the genus from Brazil and is the fourth species parasitizing nephropid lobsters in the world.

Uranium toxicity to aquatic invertebrates: A laboratory assay.

Uranium mining is an environmental concern because of runoff and the potential for toxic effects on the biota. To investigate uranium toxicity to freshwater invertebrates, we conducted a 96-h acute toxicity test to determine lethal concentrations (testing concentrations up to 262 mg L-1) for three stream invertebrates: a shredder caddisfly, Schizopelex festiva Rambur (Trichoptera, Sericostomatidae); a detritivorous isopod, Proasellus sp. (Isopoda, Asellidae); and a scraper gastropod, Theodoxus fluviatilis (Gastropoda, Neritidae). Next, we ran a chronic-toxicity test with the most tolerant species (S. festiva) to assess if uranium concentrations found in some local streams (up to 25 µg L-1) affect feeding, growth and respiration rates. Finally, we investigated whether S. festiva takes up uranium from the water and/or from ingested food. In the acute test, S. festiva survived in all uranium concentrations tested. LC50-96-h for Proasellus sp and T. fluviatilis were 142 mg L-1 and 24 mg L-1, respectively. Specimens of S. festiva exposed to 25 µg L-1 had 47% reduced growth compared with specimens under control conditions (21.5 ±â€¯2.9 vs. 40.6 ±â€¯4.9 µg of mass increase animal-1·day-1). Respiration rates (0.40 ±â€¯0.03 µg O2·h-1·mg animal-1) and consumption rates (0.54 ±â€¯0.05 µg µg animal-1·day-1; means ±â€¯SE) did not differ between treatments. Under laboratory conditions S. festiva accumulated uranium from both the water and the ingested food. Our results indicate that uranium can be less toxic than other metals or metalloids produced by mining activities. However, even at the low concentrations observed in streams affected by abandoned mines, uranium can impair physiological processes, is bioaccumulated, and is potentially transferred through food webs.

Topical Treatment of Skin Injury Inflicted in Mice by X-Ray Irradiation.

BACKGROUND/AIMS: There is no treatment, without side effects, efficiently preventing or curing skin burns, caused by radiotherapy. A new experimental topical treatment protocol was assessed in mice receiving orthovoltage X-rays at an equivalent dose to that applied to human breast cancer patients in conventional radiotherapy. METHODS: SKH-HR2 female hairless mice were irradiated on their dorsum with a total dose of 4,300 cGy during a 1-month period (20 fractions). The treatment group received a combination of 3 topical products, an oil-in-water cream, a gel containing Pinus halepensis bark aqueous extract, and an ointment containing olive oil extract of the marine isopod Ceratothoa oestroides. The positive control group was treated with a conventionally used commercial gel, whereas the negative control group did not receive any topical treatment. Skin alterations were evaluated by macroscopic examinations, measurements of transepidermal water loss (TEWL), melanin content, erythema intensity, hydration, and histopathology assessment. RESULTS: Sixty days after radiation, TEWL and hydration values were abnormal and elements of acute, chronic, and granulomatous inflammation were present in all cases. The severest damage was detected in the deeper dermis. Treatment showed a comparatively beneficial effect on chronic and granulomatous inflammation while positive control was beneficial on acute inflammation. CONCLUSION: Skin anti-inflammatory treatment was the most effective but must be applied for several months. Further preclinical studies should be conducted, assimilating a human cancer radiation therapeutic schema with the aim of optimizing skin inflammation treatment.

Evaluation of antinociceptive and anti-inflammatory effects of aqueous extract of Armadillidium vulgare Latreille.

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.

Evaluation of antinociceptive and anti-inflammatory effects of aqueous extract of Armadillidium vulgare Latreille / 中国结合医学杂志

<p><b>OBJECTIVE</b>To assess the antinociceptive and anti-inflammatory properties of the aqueous extract of Armadillidium vulgare (AV).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>

Studies on the analgesic activities of Jia-Yuan-Qing pill and its safety evaluation in mice.

The analgesic activity of Porcellio laevis Latreille, Rhizoma Corydalis, and Radix Cynanchi Paniculati have been reported in recent years. A new formula named Jia-Yuan-Qing pill (JYQP) is therefore created by combining the three herbs at 9:7:7 ratio according to traditional Chinese theories. The present study aims to evaluate the effect of JYQP as a novel painkiller in various models. Acute toxicity test was applied to evaluate the safety of JYQP. Acetic-acid-induced writhing, hot plate test, formalin test, and naloxone-pretreated writhing test were employed to elaborate the analgesic activity of JYQP and its possible mechanism. A bone cancer pain mouse model was performed to further assess the effect of JYQP in relieving cancer pain. Test on naloxone-precipitated withdrawal symptoms was conduct to examine the physical dependence of mice on JYQP. Data revealed that JYQP reduced writhing and stretching induced by acetic acid; however, this effect could not be blocked by naloxone. JYQP specifically suppressed the phase II reaction time in formalin-treated mice; meanwhile, no analgesic effect of JYQP in hot plate test was observed, indicating that JYQP exerts analgesic activity against inflammatory pain rather than neurogenic pain. Furthermore, JYQP could successfully relieve bone cancer pain in mice. No physical dependence could be observed upon long-term administration in mice. Collectively, our present results provide experimental evidence in supporting clinical use of JYQP as an effective and safe agent for pain treatment.

Environmental assessment of depleted uranium used in military armor-piercing rounds in terrestrial systems.

Depleted uranium (DU) from the military testing and use of armor-piercing kinetic energy penetrators has been shown to accumulate in soils; however, little is known about the toxicity of DU geochemical species created through corrosion or weathering. The purpose of the present study was to assess the toxic effects and bioaccumulation potential of field-collected DU oxides to the model terrestrial invertebrates Eisenia fetida (earthworm) and Porcellio scaber (isopod). Earthworm studies were acute (72 h) dermal exposures or 28-d spiked soil exposures that used noncontaminated field-collected soils from the US Army's Yuma and Aberdeen Proving Grounds. Endpoints assessed in earthworm testing included bioaccumulation, growth, reproduction, behavior (soil avoidance), and cellular stress (neutral red uptake in coelomocytes). Isopod testing used spiked food, and endpoints assessed included bioaccumulation, survival, and feeding behavior. Concentration-dependent bioaccumulation of DU in earthworms was observed with a maximum bioaccumulation factor of 0.35; however, no significant reductions in survival or impacts to cellular stress were observed. Reproduction lowest-observed-effect concentrations (LOEC) of 158 mg/kg and 96 mg/kg were observed in Yuma Proving Ground and a Mississippi reference soil (Karnac Ferry), respectively. Earthworm avoidance of contaminated soils was not observed in 48-h soil avoidance studies; however, isopods were shown to avoid food spiked with 12.7% by weight DU oxides through digital tracking studies.

Leaf litter quality drives litter mixing effects through complementary resource use among detritivores.

To comprehend the potential consequences of biodiversity loss on the leaf litter decomposition process, a better understanding of its underlying mechanisms is necessary. Here, we hypothesize that positive litter mixture effects occur via complementary resource use, when litter species complement each other in terms of resource quality for detritivores. To investigate this, monocultures and mixtures of two leaf litter species varying in quality were allowed to decompose with and without a single macro-detritivore species (the terrestrial woodlice Oniscus asellus). Resource quality of the mixture was assessed by the mean concentration, the dissimilarity in absolute and relative concentrations, and the covariance between nitrogen (N), phosphorus (P) and calcium (Ca) supply. Our results clearly show that litter mixing effects were driven by differences in their resource quality for detritivores. In particular, complementary supply of N and P was a major driver of litter mixing effects. Interestingly, litter mixing effects caused by the addition of woodlice were predominantly driven by N dissimilarity, whereas in their absence, increased P concentration was the main driver of litter mixing effects. These results show that ultimately, litter diversity effects on decomposition may be driven by complementary resource use of the whole decomposer community (i.e., microbes and macro-detritivores).

Calcium bodies of Titanethes albus (Crustacea: Isopoda): molt-related structural dynamics and calcified matrix-associated bacteria.

Crustaceans form a variety of calcium deposits in which they store calcium necessary for the mineralization of their exoskeletons. Calcium bodies, organs containing large amounts of calcium, have been reported in some terrestrial isopod crustaceans, but have not yet been extensively studied. We analyzed the architecture of these organs during the molt cycle in the isopod Titanethes albus. Two pairs of calcium bodies are positioned ventrolaterally in posterior pereonites of T. albus. Individual organs are epithelial sacs that contain material arranged in concentric layers delimited by thin laminae. As demonstrated by electron microscopy and fluorescence in situ hybridization, abundant bacteria are present within the calcium bodies. Regardless of the molt cycle stage, crystalline concretions are present in the central areas of the calcium bodies. Energy dispersive X-ray spectrometry of the concretions demonstrated that they are composed predominantly of calcium and phosphorus and selected area electron diffraction indicated the presence of hydroxyapatite. In molting animals, a glassy layer of mineralized matrix is formed between the envelope and the outermost lamina of the calcium body. This layer consists of an amorphous calcium mineral which contains less phosphorus than the central concretions and is resorbed after molt. Since changes in the mineralized matrix are synchronized with the molt cycle, the calcium bodies likely function as a storage compartment that complements sternal deposits as a source of calcium for the mineralization of the exoskeleton. Bacteria associated with the mineralized matrix of calcium bodies are evidently involved in calcium dynamics.

[Study on differentiation and extract determination of Porcellio scaber and Armadillidium vulgare].

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.

Biological reactivity of TiO2 nanoparticles assessed by ex vivo testing.

Isolated digestive gland epithelium from a model invertebrate organism was used in an ex vivo system to assess the potential of nanoparticulate TiO(2) to disrupt cell membranes. Primary particle size, surface area, concentration of particles in a suspension, and duration of exposure to TiO(2) particles were all found to have effects, which are observed at concentrations of nano-TiO(2) as low as 1 µg mL(-1). The test system employed here can be used as a fast screening tool to assess biological potential of nanoparticles with similar chemical composition but different size, concentration, or duration of exposure. We discuss the potential of ex vivo tests to avoid some of the limitations of conventional in vitro tests.

Biomonitoring of coastal areas in Tunisia: stable isotope and trace element analysis in the Yellow-legged Gull.

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.

Inhibition of multidrug/xenobiotic resistance transporter by MK571 improves dye (Fura 2) accumulation in crustacean tissues from lobster, shrimp, and isopod.

Multidrug/xenobiotic resistance transporters are present in living organisms as a first line defence system against small, potentially harmful molecules from the environment or from internal metabolic reactions. Multidrug resistance associated proteins (MRP) are one type of ATP-Binding-Cassette (ABC) transporters, which also transport dyes such as Fura 2, a calcium chelating fluorescence indicator. The specific MRP inhibitor MK571 was used to investigate the fluorescence intensity of cells in tissues of the brain and the midgut gland of the crustaceans Homarus gammarus (lobster), Crangon crangon (brown shrimp) and Idotea emarginata (isopod) during incubation with Fura 2AM (1 microM). In the presence of the inhibitor MK571 (50 microM), the fluorescence of brain tissue significantly increased in all of the three species. The midgut gland of H. gammarus showed a significant increase of fluorescence, whereas there was no effect in the midgut glands of C. crangon and I. baltica. The half maximal concentration of MK571 was 50 microM as measured in the midgut gland of H. gammarus. In conclusion, MRP transporters are present in the three investigated crustacean nervous systems. Using the midgut glands of the three species, only in H. gammarus MK571 inhibited dye extrusion, indicating species-specific differences of transporter systems, their specificity, or tissue specific expression.