Trace elements bioaccumulation in Porcellionides pruinosus (Brandt, 1833) and related biomarkers of exposure.

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.

Sugar and mineral enriched fraction from olive mill wastewater for promising cosmeceutical application: characterization, in vitro and in vivo studies.

Nowadays, agro-food by-products represent a potential low-cost source of biologically active ingredients which have been paid significant attention as nutraceuticals, medicine, food and cosmetics. In a previous study we evaluated the total sugars, metals and polyphenols of olive mill wastewater (OMWW) from a Cerasuola olive cultivar. In the present work we selectively recovered a sugar and mineral enriched fraction (SMEF) from Cerasuola OMWW by a green adsorption/desorption process. The SMEF was mainly found to be composed of monosaccharides and potassium by HPLC-ELSD and ICP-MS. The in vitro cytotoxicity on human fibroblasts, at different concentrations of the fraction, was investigated by MTT and comet assays. In addition, intracellular reactive oxygen species (ROS) production, apoptosis and cell morphological changes were examined. The physical stability of a formulation containing the SMEF (1% w/w) and its in vivo skin effects were also assessed.Our results highlighted that the SMEF showed a toxic effect at higher concentrations (i.e. cell viability reduction, DNA fragmentation and morphological alterations) well correlated with high ROS levels. Conversely, at low concentrations (0.5% and 1% w/w), no significant changes were observed. For the first time, through stability studies and in vivo tests, we also demonstrated that the SMEF formulation is stable and safe for topical application, since skin hydration improvement without negative effects was observed after 7 days of its use. Therefore, the SMEF has great potential to be used for cosmeceutical applications.

Toxic Effects of Zinc Chloride on the Bone Development in Danio rerio (Hamilton, 1822).

The increase of heavy metals in the environment involves a high exposure of aquatic organisms to these pollutants. The present study is planned to investigate the effects of zinc chloride (ZnCl2) on the bone embryonic development of Danio rerio and confirm the use of zebrafish as a model organism to study the teratogenic potential of this pollutant. Zebrafish embryos were exposed to different ZnCl2 concentrations and analyzed by ICP-MS. The skeletal anomalies were evaluated to confocal microscope after staining with calcein solution and RhodZin(TM)-3,AM. The data show a delay in hatching compared with the controls, malformations in the process of calcification and significant defects in growth. In conclusion, the current work demonstrates for the first time the Zn toxic effects on calcification process and confirm zebrafish (Danio rerio) as suitable alternative vertebrate model to study the causes and the mechanisms of the skeletal malformations.

Isopods of the genus Ligia as potential biomonitors of trace metals from the gulf of California and pacific coast of the Baja California peninsula.

Supralittoral and high intertidal coastal zones are exposed to pollution from both marine and terrestrial sources and undergo higher deposition rates than the subtidal zone. It is therefore important to identify organisms for this section of the coastal area that can be tolerant to contaminants. The aim of this study was to determine if supralittoral isopods of the genus Ligia can be used as biomonitors, since they are abundant and widely distributed. For this purpose, concentrations of trace elements were determined in Ligia isopods in toto from 26 locations across the Gulf of California and Pacific coast of the Baja California peninsula, which were collected during the summers of 2009 and 2010. The concentrations of trace elements followed the order of; Zn≥Cu>As>Cd>Pb>Hg. Elevated concentrations of copper (up to 1010 µg/g) were detected in Ligia from Santa Rosalía (SRo), a locality where industrial mining of copper has historically occurred. Industrial and municipal sewage discharges appear to have contributed to the high concentrations of zinc (326 µg/g) and lead (144 µg/g) found in organisms from Guaymas location. The high mercury concentration in organisms from Mazatlán (M) (2.01 µg/g) was associated with a thermoelectric plant. Natural sources of metals were also detected; coastal upwelling appears to be associated with high cadmium concentrations in Ligia from Punta Baja (PB) (256 µg/g) in the Pacific coast, whereas hydrothermal vents may have contributed to high concentrations of arsenic at Ensenada (E) (61 µg/g). Our results suggest that Ligia isopods reflect the natural and anthropogenic inputs of trace metals in the environment and could potentially be used as biomonitor organisms of the intertidal rocky shores of the Gulf of California and Pacific coast.

Spermiophagic activity in the female genital system of some species of terrestrial isopods (Crustacea, Halophilosciidae).

The females of some species of the family Halophilosciidae receive in the course of mating a quantity of sperm considerably redundant with respect to the number of eggs that can be fertilized; this is possible thanks to the peculiar morpho-functional organization that characterizes their genital system and that allows them to store the sperm not only in the great seminal receptacle but also within the ovary. While most of the sperm stay free in the lumen of the seminal receptacle, a part of those present in the ovary undergoes a process of capture by the follicular cells with consequent internalization within endocellular cavities. This process concerns exclusively the immotile tail, that characterizes the peculiar spermatozoon of the isopods and which is essentially of proteic nature. After their capture the sperm tails undergo a gradual process of digestion, which seems to be apparently realized without the intervention of lysosomes. The possible role of this spermiophagic activity might be to represent a significant trophic paternal investment aimed at improving the fitness of the female and of the offsprings.

Ligia italica (Isopoda, Oniscidea) as bioindicator of mercury pollution of marine rocky coasts.

In this study, we evaluated the possible role of Ligia italica as a bioindicator for the monitoring of heavy metals pollution in the suppralittoral zone of marine rocky coasts. Between 2004 and 2011 specimens of L. italica were collected along the Eastern Sicilian coasts from sites known for their high pollution levels as they are near to an area where in September 2001 a refinery plant discharged into the sea some waste containing Hg. Other specimens were collected from the Vendicari Natural Reserve located about 30 miles from the polluted sites and used as control area. On a consistent number of animals, the concentration in toto of As, Cd, Cr, Hg, Ni, Pb, V, was determined by Atomic Absorption Spectrometry. On other animals, investigations were carried out in order to check for ultrastructural alterations of the hepatopancreas, that is the main metals storage organ in isopods. Results revealed the presence, in the animals collected in 2004 from the polluted sites, of considerable concentrations of Hg and of lower concentrations of other metals such as As, Pb and V. The Hg bioaccumulation resulted in remarkable ultrastructural alterations of the two cellular types (B and S cells) in the epithelium of the hepatopancreas. Surprisingly, a moderate amount of Hg was also found in specimens collected in 2004 from the Vendicari Natural Reserve, proving that the Hg pollution can also spread many miles away. Animals collected from the polluted sites in the following years showed a progressively decreasing Hg content, reaching very low levels in those from the last sampling. Also, the ultrastructural alterations found in the hepatopancreas of the animals from the last sample were quite irrelevant. In conclusion, Ligia italica can represent a good bioindicator and the ultrastructure of the hepatopancreas could be used as ultrastructural biomarker of heavy metals pollution in the supralittoral zones.