Detection of Genetic Patterns in Endangered Marine Species Is Affected by Small Sample Sizes.

Knowledge of Genetic diversity and its spatial distribution is crucial to improve conservation plans for endangered species. Genetic tools help ensure species' long-term persistence by unraveling connectivity patterns and evolutionary trajectories of populations. Here, microsatellite genotypes of individuals from populations of Patella ferruginea are used to assess the effect of sample size on metrics of within-and between-population genetic diversity by combining empirical and simulated data. Within-population metrics are slightly to moderately affected by small sample size, albeit the magnitude of the bias is proportional to the effective population size and gene flow. The power of detecting genetic differentiation among populations increases with sample size, albeit the gain of increasing the number of sampled individuals tends to be negligible between 30 and 50. Our results line up with those of previous studies and highlight that small sample sizes are not always a hindrance to investigating genetic patterns in endangered marine species. Caution is needed in interpreting genetic patterns based on small sample sizes when the observed genetic differentiation is weak. This study also highlights the importance of carrying out genetic monitoring in seemingly well-preserved but potentially isolated populations.

Genetic patterns in Mugil cephalus and implications for fisheries and aquaculture management.

Exploitation of fisheries and aquaculture practices are exposing marine fish populations to increasing genetic risks. Therefore, the integration of genetic information into fisheries and aquaculture management is becoming crucial to ensure species' long-term persistence. The raising commercial value of grey mullet (Mugil cephalus) and its roe represents a growing challenge to the sustainable management of this economically important fishery resource. Here, microsatellites were used to investigate patterns of genetic variation in a Mediterranean area that harbor flourishing fisheries and practice semi-intensive farming of grey mullet. Genetic diversity within populations is smaller than values reported in previous studies as a result of the lower polymorphism displayed by the new microsatellite loci. Lack of genetic structuring points to the existence of a unique genetic stock, which is consistent with the species' high dispersal capabilities. Nonetheless, differences in local population effective size as well as the excess of related individuals do not completely fit the picture of a large panmictic population. Baseline genetic information here gathered will allow to set up the genetic monitoring of regional fish stocks, which is needed to assess the impact of both harvesting and aquaculture on the genetic integrity of Mugil cephalus wild populations.

Influence of genetic drift on patterns of genetic variation: The footprint of aquaculture practices in Sparus aurata (Teleostei: Sparidae).

Aquaculture finfish production based on floating cage technology has raised increasing concerns regarding the genetic integrity of natural populations. Accidental mass escapes can induce the loss of genetic diversity in wild populations by increasing genetic drift and inbreeding. Farm escapes probably represent an important issue in the gilthead sea bream (Sparus aurata), which accounted for 76.4% of total escapees recorded in Europe during a 3-year survey. Here, we investigated patterns of genetic variation in farmed and wild populations of gilthead sea bream from the Western Mediterranean, a region of long gilthead sea bream farming. We focused on the role that genetic drift may play in shaping these patterns. Results based on microsatellite markers matched those observed in previous studies. Farmed populations showed lower levels of genetic diversity than wild populations and were genetically divergent from their wild counterparts. Overall, farmed populations showed the smallest effective population size and increased levels of relatedness compared to wild populations. The small broodstock size coupled with breeding practices that may favour the variance in individual reproductive success probably boosted genetic drift. This factor appeared to be a major driver of the genetic patterns observed in the gilthead sea bream populations analysed in the present study. These results further stress the importance of recommendations aimed at maintaining broodstock sizes as large as possible and equal sex-ratios among breeders, as well as avoiding unequal contributions among parents.

Mitochondrial DNA reveals genetic structuring of Pinna nobilis across the Mediterranean Sea.

Pinna nobilis is the largest endemic Mediterranean marine bivalve. During past centuries, various human activities have promoted the regression of its populations. As a consequence of stringent standards of protection, demographic expansions are currently reported in many sites. The aim of this study was to provide the first large broad-scale insight into the genetic variability of P. nobilis in the area that encompasses the western Mediterranean, Ionian Sea, and Adriatic Sea marine ecoregions. To accomplish this objective twenty-five populations from this area were surveyed using two mitochondrial DNA markers (COI and 16S). Our dataset was then merged with those obtained in other studies for the Aegean and Tunisian populations (eastern Mediterranean), and statistical analyses (Bayesian model-based clustering, median-joining network, AMOVA, mismatch distribution, Tajima's and Fu's neutrality tests and Bayesian skyline plots) were performed. The results revealed genetic divergence among three distinguishable areas: (1) western Mediterranean and Ionian Sea; (2) Adriatic Sea; and (3) Aegean Sea and Tunisian coastal areas. From a conservational point of view, populations from the three genetically divergent groups found may be considered as different management units.

Washing treatment of automotive shredder residue (ASR).

Worldwide, the amount of end-of-life vehicles (ELVs) reaches 50 million units per year. Once the ELV has been processed, it may then be shredded and sorted to recover valuable metals that are recycled in iron and steelmaking processes. The residual fraction, called automotive shredder residue (ASR), represents 25% of the ELV and is usually landfilled. In order to deal with the leachable fraction of ASR that poses a potential threat to the environment, a washing treatment before landfilling was applied. To assess the potential for full-scale application of washing treatment, tests were carried out in different conditions (L/S = 3 and 5L/kgTS; t = 3 and 6 h). Moreover, to understand whether the grain size of waste could affect the washing efficiency, the treatment was applied to ground (<4 mm) and not-ground samples. The findings obtained revealed that, on average, washing treatment achieved removal rates of more than 60% for dissolved organic carbon (DOC), chemical oxygen demand (COD) and total Kjeldahl nitrogen (TKN). With regard to metals and chlorides, sulphates and fluoride leachable fraction, a removal efficiency of approximately 60% was obtained, as confirmed also by EC values. The comparison between the results for ground and not-ground samples did not highlight significant differences.

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.

Washing of waste prior to landfilling.

The main impact produced by landfills is represented by the release of leachate emissions. Waste washing treatment has been investigated to evaluate its efficiency in reducing the waste leaching fraction prior to landfilling. The results of laboratory-scale washing tests applied to several significant residues from integrated management of solid waste are presented in this study, specifically: non-recyclable plastics from source separation, mechanical-biological treated municipal solid waste and a special waste, automotive shredded residues. Results obtained demonstrate that washing treatment contributes towards combating the environmental impacts of raw wastes. Accordingly, a leachate production model was applied, leading to the consideration that the concentrations of chemical oxygen demand (COD) and total Kjeldahl nitrogen (TKN), parameters of fundamental importance in the characterization of landfill leachate, from a landfill containing washed wastes, are comparable to those that would only be reached between 90 and 220years later in the presence of raw wastes. The findings obtained demonstrated that washing of waste may represent an effective means of reducing the leachable fraction resulting in a consequent decrease in landfill emissions. Further studies on pilot scale are needed to assess the potential for full-scale application of this treatment.

Waste washing pre-treatment of municipal and special waste.

Long-term pollution potential in landfills is mainly related to the quality of leachate. Waste can be conveniently treated prior to landfilling with an aim to minimizing future emissions. Washing of waste represents a feasible pre-treatment method focused on controlling the leachable fraction of residues and relevant impact. In this study, non-recyclable plastics originating from source segregation, mechanical-biological treated municipal solid waste (MSW), bottom ash from MSW incineration and automotive shredder residues (ASR) were treated and the removal efficiency of washing pre-treatment prior to landfilling was evaluated. Column tests were performed to simulate the behaviour of waste in landfill under aerobic and anaerobic conditions. The findings obtained revealed how waste washing treatment (WWT) allowed the leachability of contaminants from waste to be reduced. Removal rates exceeding 65% were obtained for dissolved organic carbon (DOC), chemical oxygen demand (COD) and Total Kjeldahl Nitrogen (TKN). A percentage decrease of approximately 60% was reached for the leachable fraction of chlorides, sulphates, fluoride and metals, as proved by a reduction in electric conductivity values (70%).

Patterns of spatial genetic structuring in the endangered limpet Patella ferruginea: implications for the conservation of a Mediterranean endemic.

Patella ferruginea Gmelin, 1791 is an endangered marine gastropod endemic to the Western Mediterranean. Its range is restricted to the Sardinian-Corsican region (SCR), North Africa, a few scattered sites in Southern Spain, and Sicily. Inter-simple sequence repeat (ISSR) markers and three different mitochondrial DNA (mtDNA) regions, Cytochrome c Oxidase subunit I, 12S (small-subunit ribosomal RNA gene) and 16S (large-subunit ribosomal RNA gene), were used to investigate the presence of genetic population structuring. The mtDNA sequences showed very low levels of genetic differentiation. Conversely, ISSRs showed the presence of two main genetic groups, corresponding to Spain, North Africa and Sicily and the SCR. The SCR was further split into two subgroups. The ISSR results suggest that, on a regional scale, the genetic structure of P. ferruginea is mainly determined by the restriction of gene flow by dispersal barriers. On a more local scale human harvesting may play a crucial role in population structuring by increasing the effect of genetic drift.

Population structure of the Monocelis lineata (Proseriata, Monocelididae) species complex assessed by phylogenetic analysis of the mitochondrial Cytochrome c Oxidase subunit I (COI) gene.

Monocelis lineata consists of a complex of sibling species, widespread in the Mediterranean and Atlantic Ocean. Previous genetic analysis placed in evidence at least four sibling species. Nevertheless, this research was not conclusive enough to fully resolve the complex or to infer the phylogeny/phylogeography of the group. We designed specific primers aiming at obtaining partial sequences of the mtDNA gene Cytochrome c Oxidase subunit I (COI) of M. lineata, and have identified 25 different haplotypes in 32 analyzed individuals. The dendrogram generated by Neighbor-Joining analysis confirmed the differentiation between Atlantic and Mediterranean siblings, as well as the occurrence of at least two Mediterranean sibling species. Thus validated, the method here presented appears as a valuable tool in population genetics and biodiversity surveys on the Monocelis lineata complex.

Population structure of the Monocelis lineata (Proseriata, Monocelididae) species complex assessed by phylogenetic analysis of the mitochondrial Cytochrome c Oxidase subunit I (COI) gene

Monocelis lineata consists of a complex of sibling species, widespread in the Mediterranean and Atlantic Ocean. Previous genetic analysis placed in evidence at least four sibling species. Nevertheless, this research was not conclusive enough to fully resolve the complex or to infer the phylogeny/phylogeography of the group. We designed specific primers aiming at obtaining partial sequences of the mtDNA gene Cytochrome c Oxidase subunit I (COI) of M. lineata, and have identified 25 different haplotypes in 32 analyzed individuals. The dendrogram generated by Neighbor-Joining analysis confirmed the differentiation between Atlantic and Mediterranean siblings, as well as the occurrence of at least two Mediterranean sibling species. Thus validated, the method here presented appears as a valuable tool in population genetics and biodiversity surveys on the Monocelis lineata complex.

Heavy metal bioavailability and chelate mobilization efficiency in an assisted phytoextraction process.

The heavy metal bioavailable fraction of a soil is a core parameter to verify the potential risks of contaminant exposure to organisms or plants. The purpose of the present work is to identify the bioavailable metal fraction in soils treated with chelates. This fraction was evaluated directly by analyzing metal concentrations in soil solution and indirectly using sequential extraction procedures. The metal bioavailable fraction was compared with metal accumulated in plant leaves, grown in both untreated and chelate-treated reactors. In order to verify the effect of the readily and slowly biodegradable chelates [S,S]-ethylenediaminedisuccinic acid (EDDS), methylglycine diacetic acid (MGDA), and ethylenediaminetetraacetic acid (EDTA) on metal speciation in soils, a simulation of chelate treatment was made and metal concentrations in different soil compartments before and after the simulation were compared. Lead concentration in the soil solution was positively correlated with metal concentration in the test plants. The soluble fraction showed the best correlation with metal concentration in soil solution. The simulation of the chelate treatment demonstrated that EDTA and EDDS were able to extract part of the organic- and sulfide-bound fraction, which are less available to plants.