Detection of Viruses in Coastal Seawater Using <Emphasis Type="Italic">Mytilus Galloprovincialis</Emphasis> as an Accumulation Matrix

The purpose of this study was to demonstrate that shellfish can be used to detect enteric viruses in marine waters where they are present at very low concentrations. Aqua-cultured mussels were placed in the sea just off the mouth of a drainage channel affected by human and animal faecal contamination. Samples were taken from the channel, the sea and the mussels at intervals over two 4-week periods. The samples were tested to verify the presence of both rotaviruses and E. coli. Rotaviruses were detected by Real Time-PCR, typed by multiplex PCR and subsequently sequenced. E. coli was enumerated in water matrices by a filtering method and in mussels by the MPN method. The presence of E. coli in the examined matrices demonstrates contamination of faecal origin throughout the studied environments. Rotaviruses were recorded in channel waters, but not in sea water. In both experiments, rotaviruses were detected in mussels 21 and 28 days after being placed in the sea water off the channel mouth. The use of mussels thus enabled the detection of rotaviruses in waters where the high dilution rendered direct investigation impossible. This study indicates that mussels can be used in marine virological surveillance programs.     

Seasonality and microhabitat selection in a forest-dwelling salamander

Many small terrestrial vertebrates exhibit limited spatial movement and are considerably exposed to changes in local environmental variables. Among such vertebrates, amphibians at present experience a dramatic decline due to their limited resilience to environmental change. Since the local survival and abundance of amphibians is intrinsically related to the availability of shelters, conservation plans need to take microhabitat requirements into account. In order to gain insight into the terrestrial ecology of the spectacled salamander Salamandrina perspicillata and to identify appropriate forest management strategies, we investigated the salamander’s seasonal variability in habitat use of trees as shelters in relation to tree features (size, buttresses, basal holes) and environmental variables in a beech forest in Italy. We used the occupancy approach to assess tree suitability on a non-conventional spatial scale. Our approach provides fine-grained parameters of microhabitat suitability and elucidates many aspects of the salamander’s terrestrial ecology. Occupancy changed with the annual life cycle and was higher in autumn than in spring, when females were found closer to the stream in the study area. Salamanders showed a seasonal pattern regarding the trees they occupied and a clear preference for trees with a larger diameter and more burrows. With respect to forest management, we suggest maintaining a suitable number of trees with a trunk diameter exceeding 30 cm. A practice of selective logging along the banks of streams could help maintain an adequate quantity of the appropriate microhabitat. Furthermore, in areas with a presence of salamanders, a good forest management plan requires leaving an adequate buffer zone around streams, which should be wider in autumn than in spring.     

Combined molecular mechanics, molecular dynamics and quantum mechanical study of (+)-multifidene structure and conformation

Molecular mechanics, molecular dynamics and quantum mechanical tools have been used to explore the conformational space of (+)(3S,4S)-multifidene bioactive compounds. All the computations have been performed in the absence and presence of the solvent, simulated by the explicit introduction of a number of water molecules or by the polarized continuum model (PCM). Three conformational minima, both in vacuo and in water (simulated by PCM tool), have been confirmed at ab initio level of theory (B3LYP and MP2). All minima lie in a very narrow range of energy, and the interconversion between them is possible with a small amount of energy expense.     

Inter-simple sequence repeat markers reveal strong genetic differentiation among populations of the endangered mollusc Patella ferruginea (Gastropoda: Patellidae) from two Sardinian marine protected areas

The giant Mediterranean limpet Patella ferruginea Gmelin, 1791 is an endangered marine gastropod, whose range has progressively shrinked to few, restricted areas, due to intense human exploitation. We have studied the genetic structure of the species, in order to (1) gather information about the levels of genetic variability within and between natural populations of P. ferruginea collected in two Sardinian marine protected areas (MPAs) (Penisola del Sinis—Isola di Mal di Ventre and Isola dell’ Asinara), and (2) make an attempt to find relationships between ecological and biological attributes of the species and the genetic differentiation of the populations studied. The genetic study was carried out by means of the analysis of ten inter-simple sequence repeat (ISSR) primers on a total of 40 individuals collected at four localities. Genetic analysis evidenced (1) medium to high levels of within-population genetic variability, (2) a pattern of genetic structuring that varied with spatial scales, and (3) a strong genetic differentiation between the two MPAs. Although preliminary, these results suggest that gene flow may be present only at very small geographic scale, raising concerns on the future of the conservation of the species.     

Optimisation of a microbial bioassay for contaminated soil monitoring: bacterial inoculum standardisation and comparison with Microtox assay

This work represents the first step to set up a toxicity testing procedure and to evaluate the sensitivity of the test microorganism to several classes of environmental pollutants. First, three different techniques were employed to standardise the microbial inoculum, then two different toxicity assessment protocols have been compared: Microtox and a dehydrogenase (DHase) activity inhibition test. The main goal was the optimisation of a microbial bioassay based on the dehydrogenase activity (DHase) inhibition in Pseudomonas fluorescens bacterial strain ATCC 13525. Triphenyl tetrazolium chloride (TTC) was used as electron acceptor and its reduction produces Triphenyl formazane (TPF). The P. fluorescens DHase inhibition bioassay was investigated for being a reliable and rapid method for assessing toxicity. The optimisation of the operating conditions resulted in a repeatable bioassay. Then, P. fluorescens and Vibrio fischeri sensitivity were firstly compared by testing Zn++, one of the reference compounds for Microtox test. In addition, other compounds (Ni++, Cd++, Cu++, phenol) were also tested with both bioassays. A high statistical significance of data was obtained with the logistic curve. The present work has demonstrated that P. fluorescens is as sensitive as Microtox culture (V. fischeri), for some of the metal ions. With reference to organic compounds, the lower sensitivity of P. fluorescens to phenol makes its use difficult in organic polluted samples.     

Generation and stability of bentonite colloids at the bentonite/granite interface of a deep geological radioactive waste repository

The possible mechanisms of colloid generation at the near field/far field interface of a radioactive repository have been investigated by means of novel column experiments simulating the granite/bentonite boundary, both in dynamic and in quasi-static water flow conditions. It has been shown that solid particles and colloids can be detached from the bulk and mobilised by the water flow. The higher the flow rate, the higher the concentration of particles found in the water, according to an erosion process. However, the gel formation and the intrinsic tactoid structure of the clay play an important role in the submicron particle generation even in the compacted clay and in a confined system. In fact, once a bentonite gel is formed, in the regions where the clay is contacted with water, clay colloids can be formed even in quasi-static flow conditions. The potential relevance of these colloids in radionuclide transport has been studied by evaluating their stability in different chemical environments. The coagulation kinetics of natural bentonite colloids was experimentally studied as a function of the ionic strength and pH, by means of time-resolved light scattering techniques. It has been shown that these colloids are very stable in low saline (approximately 1 x 10(-3) M) and alkaline (pH > or = 8) waters.     

Standardization of BOD₅/COD ratio as a biological stability index for MSW

The control of biodegradable substances is the key issue in evaluating the short and long-term emission potential and environmental impact of a landfill.Aerobic and anaerobic indices, such as respirometric index (RI) and biomethane potential production (GB21), can be used in the estimation of the stability of solid waste samples.Previous studies showed different degrees of relationship between BOD₅/COD ratio compared with RI4.Aim of this study is to standardize the parameter BOD₅/COD ratio and to test the methodology under different operating conditions (dynamic or static leaching and leaching duration, 6 and 24-h) keeping constant temperature and liquid/solid ratio (L/S = 10 l/kg_TS), with the introduction of a COD fractioning method. The COD fractioning is based on the differentiation between the soluble fraction (COD_sol) and the colloidal fraction (COD_coll) using a flocculation method.The BOD₅/COD and the BOD₅/COD_sol indices are both consistent and significant and can be used as stability indices.The BOD₅/COD ratio does not seem to be influenced, for the same test duration, by the type of test, static or dynamic. In the same way the longer test duration (24-h) does not influence significantly the values of BOD₅/COD ratio. As a consequence a leaching test duration of 6-h is preferable to avoid the beginning of the hydrolysis and oxidation processes.