Impact of historical contamination on meiofaunal assemblages: The case study of the Bagnoli-Coroglio Bay (southern Tyrrhenian Sea).

The effects of contaminants on marine organisms have been documented since decades, but the long-term responses and recovery rates of benthic communities to mixtures of contaminants, several years after the cessation of industrial activities, need to be further investigated. Bagnoli-Coroglio Bay (Gulf of Naples, Tyrrhenian Sea) is a typical example of historically contaminated coastal area due to industrial activities stopped at the beginning of nineties. In the present study we carried out a fine spatial scale analysis of the distribution of meiofaunal (and nematodes) assemblages along five bathymetric transects located at increasing distance from the historical source of contamination in relation with the polycyclic aromatic hydrocarbon and heavy metal concentrations present in the sediment. Meiofaunal abundance and biomass changed widely along transects but independent from the distance from the source of contamination. Even when the contamination levels were expected to induce significant detrimental biological consequences, meiofaunal abundance and biomass were similar to those reported in unpolluted benthic coastal areas worldwide. Conversely, biodiversity in terms of meiofaunal taxa richness was generally low (range: 5-8 taxa in 12 of the overall 15 stations investigated). This was explained by the lack of sensitive groups such as ostracods, gastrotrichs and tardigrades commonly encountered in benthic coastal ecosystems, thus reflecting an overall poor/moderate environmental quality of the investigated area. Nematode (structural and functional) diversity was also low, particularly at stations characterized by higher contamination levels. At the same time, nematode species composition did not change significantly among stations suggesting a widespread effect of contaminants able to reduce the variability (i.e., turnover diversity) within the assemblages of the whole study area. Overall, our results indicate that even decades after the cessation of contaminant emissions, benthic biodiversity was affected in terms of both meiofaunal taxa and nematode species. These findings strongly reinforce the call for reducing sources of chronic pollution in marine ecosystems and provide new insights for a better understanding of the ecological recovery of historically contaminated marine environments.

Multiple human pressures in coastal habitats: variation of meiofaunal assemblages associated with sewage discharge in a post-industrial area.

Marine ecosystems are globally threatened by human activities, but some areas, such as those affected by abandoned industrial plants, show an overlap of acute and chronic impacts, which determine a considerable deterioration of their health status. Here we report the results of a research conducted on coastal sewers that discharge their loads in the highly contaminated area of Bagnoli-Coroglio (Tyrrhenian Sea, Western Mediterranean). The sampling area is characterized by heavy industrial activities (a steel plant using coal, iron and limestone) started in 1905 and ceased in 1990, which left widespread heavy metals and hydrocarbon contamination. After taking into account the potential influence of sediment grain size ranges through their inclusion as covariates in the analysis, we tested the potential impact of sewage discharge on the total abundance and multivariate structure of meiofaunal assemblages, as well as on the abundance of single taxa. The organic matter was analysed in terms of total phytopigment and biopolymeric carbon concentrations. Nematoda, Copepoda (including their nauplii), and Tardigrada were the most abundant meiofaunal taxa at all sites, but nematodes did not show a consistent pattern relative to the sewage outfalls. However, the sewer located in the historically most contaminated area showed a minimal abundance of all taxa, including nematodes, while copepods were relatively less abundant at the two southernmost sewers. Comparing the north vs. south site of the sewers, higher meiofaunal abundances were observed in the southward part, likely as a result of the local circulation. The results of this study indicate the general adaptation of meiofauna to multiple stressors (sewage discharge, superimposed to chronic industrial contamination) and its likely modulation by other local processes. They also provide relevant baseline information for future restoration interventions that would take into account the spatial variation of target organisms as needed.

Impact of offshore gas platforms on the structural and functional biodiversity of nematodes.

The Mediterranean Sea hosts hundreds of offshore gas platforms, whose activity represents a potential threat to marine ecosystems. Evidence from several studies indicates that nematodes can be highly sensitive to changes in the environmental quality. Here, we investigated the response of nematode assemblages to the presence of offshore gas platforms (located in the central Mediterranean Sea) in terms of spatial heterogeneity, structural and functional diversity. Since the effect of the investigated offshore platforms on macrofaunal assemblages were previously assessed by Terlizzi et al. (2008), the study provided also the opportunity to compare the response of different benthic compartments to the same impact related to fossil fuel extraction on marine environments. The platforms had a significant impact on nematode assemblages up to 1000 m distance from the structure. The effects were evident in term of: a) more homogeneous spatial distribution of nematode assemblages, b) increased trophic diversity of deposit feeders and c) changes in life strategies with an increase of opportunistic species in sediments closer to the platforms. Such effects seemed to be related to the dimension of the platform structures, rather than to chemical pollution or changes in food availability. These findings suggest that the platforms exert a physical alteration of the surrounding environment that is reflected by altered structural and functional traits of nematode biodiversity. The use of nematodes for monitoring the effects of the platforms only partially matched with the results obtained using macrofauna, providing further insights on potential outcomes on the functional response of marine assemblages to fossil fuel extraction.

Cluster phases of decorated micellar solutions with macrocyclic ligands.

An aqueous self-assembled micellar system (sodium dodecyl sulfate, SDS, decorated with various adhesive sites, cryptand Kryptofix 222 and crown ether 18-Crown-6 molecules) has been investigated by dynamic light scattering (DLS) and small angle x-ray scattering (SAXS) to have insights into the micellar structure, the micellar interactions, and the aggregation properties of the system. DLS demonstrates the existence of populations of aggregates in the submicrometer/micrometer range, while the Guinier analysis of the SAXS curves helps in detailing objects smaller than 30 nm. The aggregates of micelles are here named cluster phases of micelles (CPMs). Considering that SDS micelles in water do not aggregate at low concentration, it is shown that macrocyclic ligands induce the SDS micelle aggregation as a function of the concentration (i.e., investigated ligand/SDS molar ratios are 5.0, 1.5, 1.0, and 0.5) and hydrophobicity of the adhesive sites. The sizes and the percentages of the micelles and the CPMs have been monitored to test the stability and reversibility of the system. DLS results clearly show that the aggregation processes of the decorated micelles are reproducible at time intervals of the order of 1 month, while the stability may not be entirely maintained after a year. As an issue of particular relevance, the higher the ligand/surfactant molar ratio, the larger are the CPMs induced. The K222 ligand results in being more effective in promoting the micellar aggregation than 18C6 as a consequence of the different hydrophobicity.

ß-Connectin studies by small-angle x-ray scattering and single-molecule force spectroscopy by atomic force microscopy.

The three-dimensional structure and the mechanical properties of a ß-connectin fragment from human cardiac muscle, belonging to the I band, from I(27) to I(34), were investigated by small-angle x-ray scattering (SAXS) and single-molecule force spectroscopy (SMFS). This molecule presents an entropic elasticity behavior, associated to globular domain unfolding, that has been widely studied in the last 10 years. In addition, atomic force microscopy based SMFS experiments suggest that this molecule has an additional elastic regime, for low forces, probably associated to tertiary structure remodeling. From a structural point of view, this behavior is a mark of the fact that the eight domains in the I(27)-I(34) fragment are not independent and they organize in solution, assuming a well-defined three-dimensional structure. This hypothesis has been confirmed by SAXS scattering, both on a diluted and a concentrated sample. Two different models were used to fit the SAXS curves: one assuming a globular shape and one corresponding to an elongated conformation, both coupled with a Coulomb repulsion potential to take into account the protein-protein interaction. Due to the predominance of the structure factor, the effective shape of the protein in solution could not be clearly disclosed. By performing SMFS by atomic force microscopy, mechanical unfolding properties were investigated. Typical sawtooth profiles were obtained and the rupture force of each unfolding domain was estimated. By fitting a wormlike chain model to each peak of the sawtooth profile, the entropic elasticity of octamer was described.

Dielectric relaxation spectroscopy of lysozyme aqueous solutions: analysis of the δ-dispersion and the contribution of the hydration water.

The dielectric properties of lysozyme aqueous solutions have been investigated over a wide frequency range, from 1 MHz to 50 GHz, where different polarization mechanisms, at a molecular level, manifest. The dielectric relaxation spectra show a multimodal structure, reflecting the complexity of the protein-water interactions, made even more intricate with the increase of the protein concentration. The deconvolution of the spectra into their different components is not unambiguous and is generally a delicate process which requires caution. We have analyzed the whole relaxation region, on the basis of the sum of simple Debye-type relaxation functions, considering three main contributions. Particular attention has been payed to the δ-dispersion, intermediate between the ß-dispersion (rotational dynamics of the protein) and the γ-dispersion (orientational polarization of the water molecules). This intermediate contribution to the dielectric spectrum is attributed to the orientational polarization of water molecules in the immediate vicinity of the protein surface (hydration water). Our measurements clearly demonstrate that, at least at high protein concentrations, the δ-dispersion has a bimodal structure associated with two kinds of hydration water, i.e., tightly bound and loosely bound hydration water. In the concentration range investigated, the existence of a three-modal δ-dispersion, as recently suggested, is not supported, on the basis of statistical tests, by the analysis of the dielectric relaxations we have performed and a bimodal dispersion is accurate enough to describe the experimental data. The amount of the hydration water has been evaluated both from the dielectric parameters associated with the δ-dispersion and from the decrement of the loss peak of the γ-dispersion. The relative weight of tightly bound and loosely bound hydration water is briefly discussed.

Small-angle neutron scattering of percolative perfluoropolyether water in oil microemulsions.

A water in oil microemulsion system composed of water, surfactant, and oil, the latter two components of perfluoropolyether (PFPE) type, has been studied by small-angle neutron scattering (SANS) with the aim of knowing the microstructure of the system and to have an insight on the connection between microstructure characterization and percolation behavior. In fact, along the dilution line W/S = 11 of the phase diagram, dielectric spectroscopy and conductivity studies revealed a dynamic percolation process taking place approaching and above the dynamic percolation threshold, leading to a system composed of droplet clusters with percolation thresholds varying with temperature from a 0.501 volume fraction of the dispersed phase at 9.3 degrees C to 0.205 at 32.5 degrees C. The SANS experimental spectra of this work have been studied by modeling the microemulsion droplets as adhesive hard spheres. For all of the samples, the surfactant area per polar head has been also measured in the Porod region of the SANS spectra. Geometric and potential parameters as well as the osmotic pressure, the second virial coefficient, and the distance between droplets have been extracted from data as a function of droplets concentration. At low concentration, that is, below percolation thresholds, the droplets behave as hard spheres, whereas at threshold and above, adhesion changes significantly the samples. In fact, for each temperature, the measured size increases versus concentration from 30 to 50 A, and the area per polar head decreases correspondingly, suggesting that a process of dynamic fusion of droplets occurs in the system above threshold, that is, couples of droplets stick and unstick continuously with interdigitation of the surfactant tails.

Dynamic light scattering and atomic force microscopy imaging on fragments of beta-connectin from human cardiac muscle.

In order to investigate the protein folding-unfolding process, dynamic light scattering (DLS) and atomic force microscopy (AFM) imaging were used to study two fragments of the muscle cardiac protein beta-connectin, also known as titin. Both fragments belong to the I band of the sarcomer, and they are composed of four domains from I(27) to I(30) (tetramer) and eight domains from I(27) to I(34) (octamer). DLS measurements provide the size of both fragments as a function of temperature from 20 up to 86 degrees C, and show a thermal denaturation due to temperature increase. AFM imaging of both fragments in the native state reveals a homogeneous and uniform distribution of comparable structures. The DLS and AFM techniques turn out to be complementary for size measurements of the fragments and fragment aggregates. An unexpected result is that the octamer folds into a smaller structure than the tetramer and the unfolded octamer is also smaller than the unfolded tetramer. This feature seems related to the significance of the hydrophobic interactions between domains of the fragment. The longer the fragment, the more easily the hydrophobic parts of the domains interact with each other. The fragment aggregation behavior, in particular conditions, is also revealed by both DLS and AFM as a process that is parallel to the folding-unfolding transition.

Small-angle neutron scattering of mixed ionic perfluoropolyether micellar solutions.

Aqueous mixed micellar solutions of perfluoropolyether carboxylic salts with ammonium counterions have been studied by small-angle neutron scattering. Two surfactants differing in the tail length were mixed in proportions n2/n3 = 60/40 w/w, where n2 and n3 are the surfactants with two and three perfluoroisopropoxy units in the tail, respectively. The tails are chlorine-terminated. The mixed micellar solutions, in the concentration range 0.1-0.2 M and thermal interval 20-40 degrees C, show structural characteristics of the interfacial shell that are very similar to ammonium n2 micellar solutions previously investigated; thus, the physics of the interfacial region is dominated by the polar head and counterion. The shape and dimensions of the micelles are influenced by the presence of the n3 surfactant, whose chain length in the micelle is 2 A longer than that of the n2 surfactant. The n3 surfactant favors the ellipsoidal shape in the concentration range 0.1-0.2 M with a 1/2 ionization degree of n2 micelles. The very low surface charge of the mixed micelles is attributed to the increase in hydrophobic interactions between the surfactant tails, due to the longer n3 surfactant molecules in micelles. The closer packing of the tails decreases the micellar curvature and the repulsions between the polar heads, by surface charge neutralization of counterions migrating from the Gouy-Chapman diffuse layer, leading to micellar growth in ellipsoids with greater axial ratios.

Structural and functional response of meiofauna rocky assemblages to sewage pollution.

Despite meiofauna being one of the most popular tool for detecting the ecological effects of different sources of impact, the application of structured experimental designs to this benthic component is typically neglected, especially in subtidal rocky shores. In this study, an asymmetrical design was used to detect hard-bottom meiofaunal abundance and biodiversity response to sewage discharge. Sampling was carried out at a depth of 3-4 m by SCUBA diving, by means of a modified manual corer. Six replicate cores were collected at three sites (80-100 m apart), at each of three locations (one purportedly impacted [I] and two controls [Cs]). A total of 84,994 specimens were collected. The outfall affected meiofaunal assemblages in terms of taxon richness, by determining a significant reduction of the number of taxa in the disturbed location, and community structure, by causing functional changes in terms of a decrease of the abundance of nematodes and an increase of the hydrozoan component. Nematodes and syllid polychaetes showed significant lower average abundance at I. Multivariate analyses showed that both meiofaunal assemblage and syllids were significantly different at I compared with Cs. The sewage outfall also affected patterns of spatial distribution at the scale of site (100 m apart) and of replicate units (centimetres apart), both in syllids and in nematodes. Our results provide evidence that the selection of multiple controls is crucial to prevent the widespread risk of Type II error, highlighting the need of more accurate experimental designs when dealing with meiofauna.

Exo-enzymatic activities and dissolved organic pools in relation with mucilage development in the Northern Adriatic Sea.

We tested the hypothesis that the appearance of mucilage in the Northern Adriatic Sea was related with the accumulation of dissolved organic compounds released by intensive enzymatic activities and not utilized as direct substrate for microbial growth. To do this enzymatic activities and dissolved organic and inorganic pools in periods characterized by the presence of mucilage and in the same seasons but in absence of mucilage were compared. Extracellular enzymatic activities (aminopeptidase, beta-glucosidase and alkaline phosphatase), nutrient pool concentrations (total dissolved nitrogen, dissolved organic nitrogen, total dissolved phosphorus, dissolved organic phosphorus) and the biochemical composition of particulate and dissolved organic matter (in terms of proteins and carbohydrates) were determined on a monthly basis over a period of 3 years. Aminopeptidase and alkaline phosphatase activities displayed higher values in springs preceding the appearance of mucilage than in spring when no mucilage was observed. Beta-Glucosidase activity showed significantly higher values in summer periods characterized by the massive production of mucilage than in summers without mucilage events. The months preceding mucilage events were also characterized by an increase of the alkaline phosphatase to aminopeptidase activity ratio and by a significant accumulation of dissolved proteins. These findings, together with the significant increase of the DON/DOP ratio, suggest that mucilage formation is favoured by the deficiency of organic P. The present study provides compelling evidences that mucilage formation is favoured by the unbalance between organic matter mobilization by enzymatic activities and the accumulation of labile dissolved organic-N compounds.

Small-angle neutron scattering of ionic perfluoropolyether micellar solutions: role of counterions and temperature.

This paper reports a small-angle neutron scattering (SANS) characterization of perfluoropolyether (PFPE) aqueous micellar solutions with lithium, sodium, cesium and diethanol ammonium salts obtained from a chlorine terminated carboxylic acid and with two perfluoroisopropoxy units in the tail (n(2)). The counterion and temperature effects on the micelle formation and micellar growth extend our previous work on ammonium and potassium salts n(2) micellar solutions. Lithium, sodium, cesium and diethanol ammonium salts are studied at 0.1 and 0.2 M surfactant concentration in the temperature interval 28-67 degrees C. SANS spectra have been analyzed by a two-shell model for the micellar form factor and a screened Coulombic plus steric repulsion potential for the structure factor in the frame of the mean spherical approximation of a multicomponent system reduced to a generalized one component macroions system (GOCM). At 28 degrees C, for all the salts, the micelles are ellipsoidal with an axial ratio that increases from 1.6 to 4.2 as the counterion volume increases. The micellar core short axis is 13 A and the shell thickness 4.0 A for the alkali micelles, and 14 and 5.1 A for the diethanol ammonium micelles. Therefore, the core short axis mainly depends on the surfactant tail length and the shell thickness on the carboxylate polar head. The bulky diethanol ammonium counterion solely influences the shell thickness. Micellar charge and average aggregation number depend on concentration, temperature and counterion. At 28 degrees C, the fractional ionization decreases vs the counterion volume (or molecular weight) increase at constant concentration for both C = 0.1 M and C = 0.2 M. The increase of the counterion volume leads also to more ellipsoidal shapes. At C = 0.2 M, at 67 degrees C, for sodium and cesium micelles the axial ratio changes significantly, leading to spherical micelles with a core radius of 15 A, lower average aggregation number, and larger fractional ionization.

Sustainable impact of mussel farming in the Adriatic Sea (Mediterranean Sea): evidence from biochemical, microbial and meiofaunal indicators.

We have investigated the impact of a large mussel farm on the benthic environment using a battery of benthic indicators of environmental quality (including biochemical, microbial and meiofaunal parameters). These were analysed through a multi-control sampling strategy over one year. The differences across the seasons are typically higher than those between the impacted and the control stations. No effects are seen in terms of the sediment oxygen penetration and the downward fluxes (as the total mass, organic and phytopigment fluxes). The indicators based on the biochemical compositions of the sediment organic matter and the microbial parameters also show no evidence of the eutrophication process, except as a slight increase in the bacterial density in the sediments beneath the long-lines of the farm during the period of highest mussel stocks. Finally, no effects are observed in terms of the benthic faunal indicators, as the meiofaunal abundance, the community structure and the taxa richness are all indistinguishable between the farm sediments and the controls. These results show that mussel farming in the investigated system is eco-sustainable and does not significantly alter the coastal marine ecosystem, both in terms of the functioning and the trophic state. The battery of indicators selected in this study represents a useful tool for the monitoring of the potential ecological impact of mussel farms, towards guaranteeing the sustainable development of aquacultures in shallow coastal environments.

Nematode community response to fish-farm impact in the western Mediterranean.

A previous investigation on fish-farm biodeposition effects on benthos, carried out in the Gaeta Gulf (northwestern Mediterranean Sea), revealed a strong impact on meiofaunal assemblages. This study implements these findings by examining in detail the nematode assemblage and its response to organic enrichment from the start of a fish farm activity to the conclusion of the fish rearing cycle. Density, community structure and individual size were utilised for univariate (genus, trophic diversity and abundance patterns) and multivariate analyses (MDS) in order to identify the best descriptors of impact and the response of the nematode assemblages. Nematodes displayed significantly reduced densities, diversity and richness in sediments beneath fish farms. The impact of biodeposition was evident not only from structural community parameters but also in terms of functional indices. Forty-five days after starting fish farming, an increase of the nematode individual biomass was observed. MDS ordination pointed out the presence of two different nematode communities in disturbed sediments and in the control site. These results were substantiated by the analysis of the temporal changes of k-dominance curves, the maturity index and, to a lesser extent, by the index of trophic diversity. Some nematode genera were highly sensitive to biodeposition (Setosabatieria, Latronema and Elzalia) and disappeared almost completely in farm sediments, whereas other genera largely increased their dominance (Sabatieria, Dorylaimopsis and Oxystomina). This study indicates that nematodes are very sensitive to this kind of environmental disturbance. The use of simple tools, such as the k-dominance analysis and maturity index, are recommended for monitoring of aquaculture impact.

Percolation phenomenon of calcium bis(2-ethylhexyl) sulfosuccinate water-in-oil microemulsions by conductivity and dielectric spectroscopy measurements.

The sodium counterion (Na+) of the sodium bis(2-ethylhexyl) sulfosuccinate (AOT) surfactant was exchanged with calcium Ca2+ to investigate the counterion charge effect on the structure of water in normal decane microemulsions. Ohmic conductivity and dielectric permittivity measurements were performed on samples at constant water to surfactant mole ratio [water]/[Ca(AOT)(2)]=26.6. Increasing the volume fraction of the dispersed phase phi, a percolation phenomenon was observed at the constant temperature of 25 degrees C. The percolation threshold was found at phi approximately 15% by Ohmic conductivity and static dielectric permittivity studied as a function of phi, and by the frequency dependence of the complex permittivity. Critical exponents typical of the static percolation mechanism (formation of bicontinuous microemulsions) were found below and above threshold. The comparison of these results obtained for the two different counterions, Ca2+ and Na+, in AOT surfactant water in normal decane microemulsions allows detection of an important difference. The percolation below threshold is dynamic for the sodium-based microemulsions, accounting for the formation of clusters of droplets, whereas calcium-based microemulsions show a static percolation. For this system, the coalescence of droplets begins to occur below threshold at phi approximately 12%.

Investment finance: off limits for women.

PIP: In Zambia, women's access to finance is limited because 1) women lack collateral, 2) administrative practices discriminate against women, 3) women lack knowledge and information, and 4) women fail to maintain required financial records because their incomes are so low. Women have invested in areas which would be categorized as "feminine," however, including garment-making and flower-growing. The UNIP Women's League, which was the government's official body charged with ensuring gender awareness in policy-making and planning failed to survive the transition to a multiparty democracy. Some churches and nongovernmental organizations have attempted to promote gender-aware projects to help women survive the current economic crisis, but these programs are few and far between and suffer from a lack of capital. Organizations must adopt a business-like approach to help women move from welfare to economic empowerment.^ieng