Genetic differentiation between morphotypes in the Antarctic limpet <Emphasis Type="Italic">Nacella concinna</Emphasis> as revealed by inter-simple sequence repeat markers

The limpet Nacella concinna (Strebel 1908) was the focus of numerous studies dealing with Antarctic benthos. One of the main characteristics of the species is the presence of two distinguishable morphotypes, one inhabiting the intertidal (during summer) and the other inhabiting the subtidal. For a long time these forms were considered as an expression of phenotypic plasticity, since previous studies did not found genetic differences between them. In the present work, we performed both a morphometric and a genetic differentiation analysis (using ISSR-PCR markers) of these two forms in three stations sampled at Potter Cove, South Shetland Islands. The results confirmed the morphological differences between intertidal and subtidal forms reported in other Antarctic localities. The genetic differences detected indicate that the two forms can be considered as genetically distinct populations maintaining low levels of gene flow. The degree of reproductive isolation of the ecotypes is discussed, as well as the possible origin of the divergence. The genetic differentiation observed can also have behavioral and physiological correlates, pointing out the importance of taking into account the potential differences in the response of both populations to different conditions in future studies in this species. M. C. de Aranzamendi and R. Sahade contributed equally to this paper.     

Effects of ranitidine and its photoderivatives in the aquatic environment

This study was designed to assess the overall ecotoxicity of ranitidine, a histamine H₂-receptor antagonist that inhibits stomach acid production. Hence, in addition to ranitidine, its main two photoderivatives, obtained by solar simulator irradiation in water, were investigated. The photoproducts were identified by their physical features. Bioassays were performed on rotifers and microcrustaceans to assess acute and chronic toxicity, while SOS Chromotest and Ames test were utilized to detect the genotoxic potential of the investigated compounds. The results showed that ranitidine did not show any acute toxicity at the highest concentration tested (100 mg/L) for all the organisms utilized in the bioassays. Chronic exposure to these compounds caused inhibition of growth population on rotifers and crustaceans. Genotoxic and mutagenic effects were especially found for one photoproduct suggesting that transformation products, as frequently demonstrated, may show effects higher than the respective parental compound.     

Blends of Poly (Butylene Adipate-Co-Terephthalate) and Thermoplastic Whey Protein Isolate: A Compatibilization Study

Journal of Polymers and the Environment - This work assesses the influence of the plasticizer polyethylene glycol (PEG) on the compatibilization of poly (butylene adipate-co-terephthalate) (PBAT)...     

A method for measuring low-weight carboxylic acids from biosolid compost

Concentration of low-weight carboxylic acids (LWCA) is one of the important parameters that should be taken into consideration when compost is applied as soil improver for plant cultivation, because high amounts of LWCA can be toxic to plants. The present work describes a method for analysis of LWCA in compost as a useful tool for monitoring compost quality and safety. The method was tested on compost samples of two different ages: 3 (immature) and 6 (mature) months old. Acids from compost samples were extracted at high pH, filtered, and freeze-dried. The dried sodium salts were derivatized with a sulfuric acid-methanol mixture and concentrations of 11 low-weight fatty acids (C1-C10) were analyzed using headspace gas chromatography. The material was analyzed with two analytical techniques: the external calibration method (tested on 11 LWCA) and the standard addition method (tested only on formic, acetic, propionic, butyric, and iso-butyric acids). The two techniques were compared for efficiency of acids quantification. The method allowed good separation and quantification of a wide range of individual acids with high sensitivity at low concentrations. Detection limit for propionic, butyric, caproic, caprylic, and capric acids was 1 mg kg(-1) compost; for formic, acetic, valeric, enanthoic and pelargonic acids it was 5 mg kg(-1) compost; and for iso-butyric acid it was 10 mg kg(-1) compost. Recovery rates of LWCA were higher in 3-mo-old compost (57-99%) than in 6-mo-old compost (29-45%). In comparison with the external calibration technique the standard addition technique proved to be three to four times more precise for older compost and two times for younger compost. Disadvantages of the standard addition technique are that it is more time demanding and laborious.     

A proteomic approach to the study of the marine mussels Mytilus edulis and M. galloprovincialis

In this study, a proteomic approach was applied for the generation of reference maps and subsequently to detect, quantify and compare the global protein expression between two related species of marine mussels, Mytilus edulis and Mytilus galloprovincialis, growing in their own geographical habitats. A comparative study of the protein profiles generated from analytical two-dimensional electrophoresis gels was performed, and changes in protein expression were analysed quantitatively by computer analysis. An average of 1,278 spots per gel was detected in 16 individuals (8 M. edulis and 8 M. galloprovincialis); however, not all spots were included in the study. Expression of 420 spots was compared, and significant differences in the intensity levels were detected in 37 protein spots (8.8%). Fifteen proteins showed higher expression in M. edulis, and 22 proteins, in M. galloprovincialis. The technique of peptide mass fingerprinting using MALDI-TOF (matrix-assisted laser desorption ionisation/time-of-flight) and/or nanoelectrospray double subfragmentation mass spectrometry enabled the unambiguous identification of 15 of these 37 differentially expressed proteins. Most of the identified proteins can be grouped basically into four broad functional classes: cytoskeletal and myofibrillar proteins, proteins associated with stress response, proteins associated with the storage or production of energy, and proteins related to rearrangement in the synthesis of native structures. These results expand our understanding of the molecular differentiation of the two mussel taxa and serve as a useful base for future ecological, physiological and genetic studies.     

Organic compounds in tire particle induce reactive oxygen species and heat-shock proteins in the human alveolar cell line A549

Debris produced from the attrition of tires of motor vehicles constitutes 5-7% of the atmospheric particulate matter (PM10). Debris particles are indeed small enough to enter human lung and thus morphological and chemical characterization has been performed. We demonstrated that the organic fraction of tire debris induces a dose-dependent increase in cell mortality, DNA damage, as well as a significant modification of cell morphology at the dose of 60 microg/ml, which may correspond to the quantity present in the air humans inhale daily. The present research aims at investigating if reactive oxygen species (ROS) production and Hsp70 expression are involved in the cascade of toxic effects produced on the A549 cell line, as it has been suggested for the ultrafine atmospheric particles and diesel exhaust. To this end, cells were exposed at the doses of 10, 50, 60, 75 microg/ml of TD organic extract (TDOE) and analyzed at different exposure time. ROS were detected by the oxidation of 2'7'-dichlorodihydrofluorescein diacetate to dichlorofluorescein, and fluorescence was measured by flow cytometry. Hsp70 protein expression was determined by immunochemical analysis, and protein expression quantification performed by optical densitometry. ROS production was analysed after 2 h of treatment. A statistically significant increase in fluorescence was observed and the intensity of the stress response was parallel to the increasing concentrations used. An evident increase of Hsp70 expression at lower doses (10, 50 microg/ml) and at longer exposure times (72 h) was observed, during the time that our previous studies showed that cell viability, plasma membrane integrity, and DNA molecules were not affected. Thus it can be deduced that the increase in Hsp70 expression protected the cells from those damages, which became evident at the higher doses, and that this parameter might be used as a sensitive indicator of exposure. These data suggest that ROS production may be the first event caused by A549 exposure to TDOE and this result is in line with other evidences provided for the role of ROS generation in ultrafine PM toxicity. It can be suggested that this event induces an overexpression of Hsp70 only at the lower doses and longer exposure time, when cells still appear unaffected. Subsequently when ROS generation reaches high levels, a general inhibition of protein synthesis probably occurs, culminating in cell toxicity.     

In situ monitoring of urban air in Southern Italy with the Tradescantia micronucleus bioassay and semipermeable membrane devices (SPMDs)

This study was designed to assess toxic and genotoxic compounds in the urban air of Caserta, South Italy using cuttings from the plant Tradescantia #4430. In situ monitoring of gaseous pollutans was made at 17 sampling points in two seasons of the year. Genotoxicity was evaluated by recording the micronuclei in meiotic pollen mother cells (Trad-MCN assay). In addition, the passive sampler semipermeable membrane devices (SPMDs) were deployed at the sampling points with a significant increase in micronuclei frequency. SPMDs concentrated priority organic pollutants were identified by high performance liquid chromatography and gas chromatography, while toxicity and mutagenesis were assessed on the bacterium Vibrio fischeri using the Microtox and Mutatox systems respectively. Significant toxic and mutagenic effects were observed at different points on the town grid and SPMDs effectively concentrated trace contaminants. The relationship between what was present in the air sampled by SPMDs and the micronuclei frequency was also explored.     

Arsenic accumulation and physiological attributes of spinach in the presence of amendments: an implication to reduce health risk

The current study examined the effect of calcium (Ca) and ethylenediaminetetraacetic acid (EDTA) on arsenic (As) uptake and toxicity to spinach (Spinacia oleracea) as well as assessed the potential human health risks. Spinach seedlings were exposed to three levels of As (25, 125, and 250 μM) alone or together with three levels of EDTA (25, 125, and 250 μM) and Ca (1, 5, and 10 mM). The effect of EDTA and Ca was assessed in terms of As contents in roots and shoots, hydrogen peroxide production, chlorophyll contents, and lipid peroxidation. The accumulation and toxicity of As to spinach plants increased with increasing As levels in nutrient solution. Exposure to As resulted in lipid peroxidation and reduced chlorophyll contents. The highest level of As alone (250 μM) showed highest human health risk (hazard quotient of 7.09 at As-250). Addition of EDTA enhanced As accumulation by spinach, while reduced As toxicity to spinach, as well as human health risk (hazard quotient of 4.01 at As-250). Similarly, Ca significantly reduced As toxicity to spinach and the human health risks (hazard quotient of 3.79 at As-250) by reducing its accumulation in spinach. Higher levels of Ca were more effective in reducing As uptake and toxicity as well as enhancing chlorophyll contents.