Effects of Heavy Metals Artificial Contamination on Porcellio laevis (Latreille, 1804) (Crustacea: Isopoda: Oniscidea).

This study aimed at determining the competition of cadmium (Cd), lead (Pb), zinc (Zn) and copper (Cu) on their assimilation, on the food consumption and the growth of terrestrial isopod Porcellio laevis. Individuals were exposed to artificially contaminated litter of Quercus for 4 weeks and were weekly weighed. At the end of the experiment, the concentration of Cd, Pb, Zn and Cu in individuals were measured by atomic absorption spectrometry. Biological parameters such as growth, and bioaccumulation factor (BAF) were calculated and results from the various treatments were compared. Depending on metals, weight loss or gain were recorded for isopods during the four weeks of exposure. A weight loss was measured on individuals exposed to Cd-contaminated litter whereas a weight gain was highlighted for those exposed to the Zn-contaminated litter. BAF values revealed that P. laevis was macroconcentrator of Zn and Cu and deconcentrator of Cd and Pb.

Measure of environmental stress on Porcellio laevis Latreille, 1804 sampled near active Tunisian industrial areas.

This study aimed to observe the type of asymmetry exhibited by Porcellio laevis sampled from 15 sites belonging to Tunisian industrialized areas. Physicochemical parameters such as pH, organic matter and CaCO3 contents were measured in soils. Moreover, Cd, Pb, Zn, and Cu concentrations were determined in both soils and woodlice. Additionally, 10 metrical traits were measured to evaluate the type of asymmetry on individuals: the basis, the second and the third articles of the antenna, the first article of the flagellum of the antenna and the merus, the carpus, and the propodus of the sixth and the seventh pereopods. Among the 531 measured individuals, 432 exhibited fluctuating asymmetry (FA) while the remaining individuals exhibited antisymmetry or directional asymmetry. The data obtained were analyzed using a multivariate statistical analysis. Contrary to our hypothesis, the results showed that individuals from contaminated sites have a low FA level, whereas those from uncontaminated sites have a high FA level, particularly females but with some exceptions. Variations in FA level in the traits and populations studied and its usefulness as a stress indicator were discussed.

Hierarchical organization of the cuticle of the subsocial desert isopod, Hemilepistus reaumurii.

The crustacean cuticle is a hierarchically organised material which provides protection and sites for muscle attachment. The physical properties of this exoskeleton envelope are adapted to the function and the eco-physiological requirements of the species. This paper aimed to study, using the TEM, the structure of the tubercle and the tergite cuticle of the arid species Hemilepistus reaumurii in a comparison with a subhumid isopod in order to relate some peculiar features to an adaptive process to environmental constraints. Results showed that wild H. reaumurii cuticles were twice as thick in comparison with Porcellio variabilis which is a subhumid zone isopod. It is suggested therefore that the thick cuticle of wild H. reaumurii can be an adaptation to terrestrial life and a protection against osmotic stress and water loss in an arid environment. In addition the inside of the tubercle showed a high number of lipid droplets stacked into an adipose tissue which suggest that tubercles were used for storage for nutritive material in wild H. reaumurii.

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.

Physiological and behavioral responses of Orchestia gammarellus (Amphipoda, Talitridae) towards trace elements contamination soil.

Effects of trace elements on the hepatopancreas ultrastructure and on the locomotor activity rhythm were investigated in the amphipod talitrid Orchestia gammarellus collected from Bizerte lagoon banks situated in the north of Tunisia. Animals were exposed to a series of contaminated soil with different concentrations of cadmium and zinc. The locomotor activity rhythm was studied under constant darkness. Histopathological analysis showed that the trace elements, especially the Cd, induced significant changes in the morphology and in the ultrastructural organization of hepatopancreatic cells. The significant alterations obtained were dose dependent. Concerning the behavioral response, results revealed the presence of two components whatever the experimental conditions. In addition, a great inter-individual variability of the locomotor rhythm was observed. Patterns were in majority bimodal for the control individuals and became unimodal and multimodal when exposed under Cd and Zn, respectively. Furthermore, ultradian and circadian periods were determined. The circadian period lengthened after Zn exposure. In addition, the locomotor activity rhythm was more stable for control individuals. However, those exposed to Cd were less active.

The induced damage in the hepatopancreas of Orchestia species after exposure to a mixture of Cu/Zn-An ultrastructural study.

The hepatopancreas of crustaceans species has been recognized as an essential target organ to assess trace elements' effects. Due to its dynamic and capability of detoxifying trace metal, this organ often indicates distinct pathological disturbances. In the present work, we intend to evaluate the bioaccumulation of trace metal in three Orchestia species (Orchestia montagui, Orchestia gammarellus, and Orchestia mediterranea) living in symmetry in the banks of Bizerte lagoon (37°13'8″N 09°55'1″E) after their exposure during 14 days to a mixture of copper and zinc, and to highlight the effect of these metals on their hepatopancreas ultrastructure using transmission electron microscopy. At the end of the experiment, results showed that the mortality and the body mass varied according to the used nominal concentrations. Significant alterations were noted in all the treatment groups. The degree of these alterations depends on the used concentration, and they are represented especially by the cells remoteness and the border lyses, the reduction of the nuclear volume, the increase in the cytoplasm density with the presence of trace metal in the nucleus as well as in the vacuole, the disorganization and the destruction of microvilli, the condensation of the majority of cellular organelles and mitochondria swelling. Through this study, Orchestia genus could be an attractive candidate for the biochemical study of trace metal toxicity in Tunisian wetlands.

Physiological and histopathological responses of Porcellio laevis (Isopoda, Crustacea) as indicators of metal trace element contamination.

This study was designed to assess the impact of the mixture of cadmium (Cd) and zinc (Zn) on the bioaccumulation and the ultrastructural changes in the hepatopancreas of Porcellio laevis (Latreille, 1804) after 4 weeks of exposure to contaminated Quercus leaves under laboratory conditions. For each metal, four concentrations were used with four replicates for each concentration. Metal concentrations in the hepatopancreas and the rest of the body were determined using atomic absorption spectrometry. From the first week until the end of the experiment, a weight gain in P. laevis was observed particularly between the first and the end of exposure from 93.3 ± 18.22 mg fw to 105.22 ± 16.16 mg fw and from 106.4 ± 22.67 mg fw to 125.9 ± 23.9 mg fw for Mix1 and Mix4, respectively. Additionally, the determined metal trace elements (MTE) concentrations in the hepatopancreas were considerably higher compared to those in the rest of the body and seem to be dose-dependent. Using transmission electron microscopy (TEM), some alterations were highlighted in the hepatopancreas. The main observed alterations were (a) the destruction of the microvilli border in a considerable portion of cells, (b) the increase of the lipid droplets with different shapes and sizes, (c) the increase in the number of the mitochondria, and (d) the appearance of TE in the form of B-type granules. The obtained results confirmed the ability of P. laevis to deal with high amounts of MTE, suggesting its possible use in future soil's biomonitoring programs.

Evaluation of trace element contamination using Armadillo officinalis Duméril, 1816 (Crustacea, Isopoda) as a tool: An ultrastructural study.

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.

Effects of trace metal elements on ultrastructural features of hepatopancreas of Armadillidium granulatum Brandt, 1833 (Crustacea, Isopoda).

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.

Ecotoxicological effects of trace element contamination in talitrid amphipod Orchestia montagui Audouin, 1826.

This study deals with the evaluation of trace element bioaccumulation and histological alterations in the hepatopancreas of the supralittoral amphipod Orchestia montagui Audouin, 1826 due to the exposure to cadmium, copper, and zinc. Orchestia montagui individuals were maintained during 14 days in soils contaminated with different trace elements namely cadmium, copper, and zinc; a control was also prepared. Our results show that the mortality and the body mass vary according to the metal and the nominal concentration used. In general, the mortality increases from the seventh day. However, the body mass shows a decrease with cadmium exposure and an increase with copper and zinc exposures. Furthermore, the concentration factor highlights that this species is considered a macroconcentrator for copper and zinc. The hepatopancreas of unexposed and exposed animals were compared to detect histological changes. Our results show significant alterations in the hepatopancreas of the exposed animals after the experiment. The degree of these alterations was found to be dose-dependent. Among the histological changes in the hepatopancreas in O. montagui, a loss of cell structure was noted, especially cell remoteness and border lyses, the reduction of nuclear volume, an increase in the cytoplasm density with the presence of trace element deposits in both the nucleus and vacuoles, a disorganization and destruction of microvilli, and a condensation of the majority of cell organelles and mitochondria swelling. Through this study, we have confirmed that O. montagui can be a relevant model to assess trace metal element pollution in Tunisian coastal lagoons with the aim of using it in future biomonitoring programs.

Biodiversity of Talitridae family (Crustacea, Amphipoda) in some Tunisian coastal lagoons.

BACKGROUND: Although wetlands were remarkable habitats with their fauna and flora diversity, few studies have been devoted to the study of amphipod distribution in this type of environment. To study both qualitatively and quantitatively amphipod community, surveys were conducted during the spring season in ten coastal lagoons ranging from subhumid to arid bioclimatic stage. At each station, eight quadrats of 50 × 50 cm were randomly placed. Amphipods were preserved in alcohol 70°C. In the laboratory, the specimens collected were identified and counted. Meanwhile, analyses of organic matter, particle size, and heavy metals from the soil taken from each station were made. RESULTS: A total of 1,340 specimens of amphipods were collected, and eight species belonging to Talitridae family were identified. Species richness ranges from one species collected in the supralittoral zone of El Bcherliya (Ghar El Melh lagoon) and eight species in the supralittoral zone of Bizerte lagoon. In this last station, the relative abundance of amphipods was significantly higher (36.04%, N = 483). In addition, the diversity indices of Simpson, Shannon-Weaver, and equitability shows that the highest species diversity characterizes this same station while the community was more balanced in opposite El Boughaz (Ghar El Melh lagoon) (J″ =0.996). CONCLUSIONS: Thespatial distribution of different amphipod species depends on edaphic (heavy metals, granulometry, organic matter) and climatic (temperature, humidity) factors.

[Reproductive phenology of a natural population of Armadillidium pelagicum Arcangeli, 1955 (Isopoda, Oniscidea)]. / Phénologie de la reproduction d'une population naturelle de Armadillidium pelagicum Arcangeli, 1955 (Isopoda, Oniscidea).

Armadillidium pelagicum, the most abundant species of the genus in Tunisia is endemic to the North of Tunisia and the circumsicilian islands. In the laboratory and under the natural conditions of temperature and photoperiod, virgin females of A. pelagicum exhibit a spontaneous ovarian maturation, followed by a parturial moult. Nevertheless, the onset of reproduction is greatly accelerated by the presence of a male. In fact, mating shortened the lag time (from the beginning of experiment to the parturial moult) by reducing the number of normal moult preceding the first parturial moult and the duration of the preparturial intermoult. In this species, only one mating is enough to ensure the fertility for several egg layings. Otherwise, it has been shown that A. pelagicum has a reproductive period rather than a reproductive activity and that mating females have a longer reproductive period than virgins. Like several species of terrestrial Isopods, the duration of the gestation period is temperature-dependent.

Towards a characterization of the locomotor activity rhythm of the supralittoral isopod Tylos europaeus.

Freshly collected samples of Tylos europaeus from Korba beach (northeast of Tunisia) were housed in an environmental cabinet at controlled temperature (18°C ± .5°C) and photoperiod. Locomotor activity was recorded under two photoperiodic regimens by infrared actography every 20 min by multichannel data loggers. One regimen simulated the natural light-dark cycle on the day of collection, whereas the second imposed a state of continuous darkness on all individuals. Under entraining conditions, the animals displayed rhythmic activity, in phase with the period of darkness, whereas in continuous darkness these isopods exhibited a strong endogenous rhythm with circadian and semidiurnal components at mean periods of τ (h:min) = 25:09 ± 01:02 h and τ = 12:32 ± 00:26 h, respectively. Under free-running conditions, this endogenous rhythm showed significant intraspecific variability.