Impact of trace metals from past mining on the aquatic ecosystem: a multi-proxy approach in the Morvan (France).

This study seeks to determine to what extent trace metals resulting from past mining activities are transferred to the aquatic ecosystem, and whether such trace metals still exert deleterious effects on biota. Concentrations of Cd, Cu, Pb and Zn were measured in streambed sediments, transplanted bryophytes and wild brown trout. This study was conducted at two scales: (i) the entire Morvan Regional Nature Park and (ii) three small watersheds selected for their degree of contamination, based on the presence or absence of past mining sites. The overall quality of streambed sediments was assessed using Sediment Quality Indices (SQIs). According to these standard guidelines, more than 96% of the sediments sampled should not represent a threat to biota. Nonetheless, in watersheds where past mining occurred, SQIs are significantly lower. Transplanted bryophytes at these sites consistently present higher trace metal concentrations. For wild brown trout, the scaled mass and liver indices appear to be negatively correlated with liver Pb concentrations, but there are no obvious relationships between past mining and liver metal concentrations or the developmental instability of specimens. Although the impact of past mining and metallurgical works is apparently not as strong as that usually observed in modern mining sites, it is still traceable. For this reason, past mining sites should be monitored, particularly in protected areas erroneously thought to be free of anthropogenic contamination.

Impact of historical mining assessed in soils by kinetic extraction and lead isotopic ratios.

The aim of this study is to estimate the long-term behaviour of trace metals, in two soils differently impacted by past mining. Topsoils from two 1 km(2) zones in the forested Morvan massif (France) were sampled to assess the spatial distribution of Cd, Cu, Pb and Zn. The first zone had been contaminated by historical mining. As expected, it exhibits higher trace-metal levels and greater spatial heterogeneity than the second non-contaminated zone, supposed to represent the local background. One soil profile from each zone was investigated in detail to estimate metal behaviour, and hence, bioavailability. Kinetic extractions were performed using EDTA on three samples: the A horizon from both soil profiles and the B horizon from the contaminated soil. For all three samples, kinetic extractions can be modelled by two first-order reactions. Similar kinetic behaviour was observed for all metals, but more metal was extracted from the contaminated A horizon than from the B horizon. More surprising is the general predominance of the residual fraction over the "labile" and "less labile" pools. Past anthropogenic inputs may have percolated over time through the soil profiles because of acidic pH conditions. Stable organo-metallic complexes may also have been formed over time, reducing metal availability. These processes are not mutually exclusive. After kinetic extraction, the lead isotopic compositions of the samples exhibited different signatures, related to contamination history and intrinsic soil parameters. However, no variation in lead signature was observed during the extraction experiment, demonstrating that the "labile" and "less labile" lead pools do not differ in terms of origin. Even if trace metals resulting from past mining and metallurgy persist in soils long after these activities have ceased, kinetic extractions suggest that metals, at least for these particular forest soils, do not represent a threat for biota.

Wild brown trout affected by historical mining in the Cévennes National Park, France.

In the protected area of the Cévennes National Park (Southern France), 114 wild brown trout (Salmo trutta fario) were captured at six locations affected to different extents by historical mining and metallurgy dating from the Iron Age to Modern Times. Cadmium and lead in trout livers and muscles reflect high sediment contamination, although an age-related effect was also detected for hepatic metal concentrations. Lead isotope signatures confirm exposure to drainage from mining and metallurgical waste. Developmental instability, assessed by fluctuating asymmetry, is significantly correlated with cadmium and lead concentrations in trout tissues, suggesting that local contamination may have affected fish development. Nowadays, the area is among the least industrialized in France. However, our results show that 60% of the specimens at one site exceed EU maximum allowed cadmium or lead concentration in foodstuffs. The mining heritage should not be neglected when establishing strategies for long-term environmental management.

On hidden heterogeneity in directional asymmetry--can systematic bias be avoided?

Directional asymmetry (DA) biases the analysis of fluctuating asymmetry (FA) mainly because among-individual differences in the predisposition for DA are difficult to detect. However, we argue that systematic bias mainly results from predictable associations between signed right-left asymmetry and other factors, i.e. from systematic variation in DA. We here demonstrate methods to test and correct for this, by analysing bilateral asymmetry in size and shape of an irregular sea urchin. Notably, in this model system, DA depended significantly on body length and geographic origin, although mean signed asymmetry (mean DA) was not significant in the sample as a whole. In contrast to the systematic variation in DA, undetectable, random variability in the underlying DA mainly leads to reduced statistical power. Using computer simulations, we show that this loss of power is probably slight in most circumstances. We recommend future studies on FA to routinely test and correct for not only as yet for mean DA, but also for systematic variation in DA.

When morphometry meets genetics: inferring the phylogeography of Carabus solieri using Fourier analyses of pronotum and male genitalia.

Population differentiation is a crucial step in the speciation process and is therefore a central subject in studies of microevolution. Assessing divergence and inferring its dynamics in space and time generally require a wide array of markers. Until now however, most studies of population structure are based on molecular markers and those concerning morphological traits are more scarce. In the present work, we studied morphological differentiation among populations of the ground beetle Carabus solieri, and tested its congruence with genetic population structure. The shape of pronotum and aedeagus was assessed using Dual Axis Fourier Shape Analysis. manova on Fourier coefficients revealed highly significant morphological variation between populations and a similar geographical pattern of differentiation for both structures. On the whole, morphological and genetic patterns were also found to be congruent. Our analysis confirms the phylogeographical scenario proposing that two entities of C. solieri differentiated during the last glaciation events before recolonizing the actual range of the species. It also indicates a large introgression between the two differentiated entities in the centre of the range.

Isolation by distance and sharp discontinuities in gene frequencies: implications for the phylogeography of an alpine insect species, Carabus solieri.

Analysis of genetic isolation by distance (IBD) is of prime importance for the study of processes responsible for spatial population genetic structure and is thus frequently used in case studies. However, the identification of a significant IBD pattern does not necessarily imply the absence of sharp discontinuities in gene frequencies. Therefore, identifying barriers to gene flow and/or secondary contact between differentiated entities remains a major challenge in population biology. Geographical genetic structure of 41 populations (1080 individuals) of an alpine insect species, Carabus solieri, was studied using 10 microsatellite loci. All populations were significantly differentiated and spatially structured according to IBD over the entire range. However, clustering analyses clearly identified three main clusters of populations, which correspond to geographical entities. Whereas IBD also occurs within each cluster, population structure was different according to which group of populations was considered. The southernmost cluster corresponds to the most fragmented part of the range. Consistently, it was characterized by relatively high levels of differentiation associated with low genetic diversity, and the slope of the regression of genetic differentiation against geographical distances was threefold those of the two other clusters. Comparisons of within-cluster and between-cluster IBD patterns revealed barriers to gene flow. A comparison of the two approaches, IBD and clustering analyses, provided us with valuable information with which to infer the phylogeography of the species, and in particular to propose postglacial colonization routes from two potential refugia located in Italy and in southeastern France. Our study highlights strongly the possible confounding contribution of barriers to gene flow to IBD pattern and emphasizes the utility of the model-based clustering analysis to identify such barriers.

Parasitic infection and developmental stability: fluctuating asymmetry in Gammarus pulex infected with two acanthocephalan species.

Several studies have reported a negative association between developmental stability and parasitic infection. However, the host-parasite associations examined so far consist only of a limited number of parasite taxa, and developmental stability was appraised on definitive hosts. The present study examines the association between infection by 2 acanthocephalan parasites. Pomphorhynchus laevis and Polymorphus minutus, and the developmental stability of their common intermediate host Gammarus pulex. Developmental stability was estimated from the fluctuating asymmetry (FA) levels of 6 morphological traits. A positive association was found between FA and infection. Infected gammarids tended to be more asymmetrical than the noninfected ones for an index generated by combining FA scores from 2 characters out of the 6 studied, even though no significant relationships were found between FA levels and parasitic loads. The simultaneous presence of both acanthocephalan species in the same host seems to be associated with increased FA levels of gammarids, but this trend was not statistically significant. For the same characters, males exhibited higher levels of FA than females.

Independence between developmental stability and canalization in the skull of the house mouse.

The relationship between the two components of developmental homeostasis, that is canalization and developmental stability (DS), is currently debated. To appraise this relationship, the levels and morphological patterns of interindividual variation and fluctuating asymmetry were assessed using a geometric morphometric approach applied to the skulls of laboratory samples of the house mouse. These three samples correspond to two random-bred strains of the two European subspecies of the house mouse and their F1 hybrids. The inter- and intraindividual variation levels were found to be smaller in the hybrid group compared to the parental ones, suggesting a common heterotic effect on skull canalization and DS. Both buffering mechanisms might then depend on the same genetic condition, i.e. the level of heterozygosity. However, related morphological patterns did not exhibit any congruence. In contradiction with previous studies on insect wing traits, we therefore suggest that canalization and DS may not act on the same morphological characters. The fact that this discrepancy could be related to the functional importance of the symmetry of the characters under consideration is discussed in the light of our knowledge of the genetic bases of both components of developmental homeostasis.

Chromosomal transmission bias in laboratory hybrids between wild strains of the two European subspecies of house mice.

Laboratory crosses between wild strains of the two European house mouse subspecies Mus musculus domesticus (2n = 34) and M. m. musculus (2n = 40) were performed to analyze the selective processes involved in the non-introgression of centromeric regions of Robertsonian (Rb) fusions in the Danish hybrid zone. The chromosomal analysis of 226 backcross progeny from 22 reciprocal crosses showed that the segregation of the three Rb fusions present did not significantly differ from Mendelian expectations. However, a significant negative correlation was found between Rb transmission rates and the average litter sizes of the F1 pairs. Among the different models of selection discussed, the most likely one supported the existence of two opposing selective factors resulting in an overall compensation of chromosomal types in the backcross progeny. A two-phase selective process involving embryo competition was postulated with non-Rb carriers being favored during pre-implantation but disadvantaged after implantation. Such balanced selective pressures acting on musculus non-Rb centromeres are compatible with the steep slope and off-centered position of the chromosomal cline observed in the Danish hybrid zone. These results suggested that these selective factors may be more related to centromere origin (musculus or domesticus) than to centromere structure (Rb or non-Rb).

Fluctuating asymmetry in the Mus musculus hybrid zone: a heterotic effect in disrupted co-adapted genomes.

Developmental stability reflects the organism's ability to buffer minor developmental accidents and is often estimated by measuring the fluctuating asymmetry. Either implicitly or explicitly, numerous authors have assumed that developmental stability is correlated with overall fitness. If this is the case, changes in morphological asymmetry across a hybrid zone could be used as a measure of the selection on hybrid genomes. Developmental stability in hybrid populations is theoretically related to the genetic distance between hybridizing taxa, and results from a balance between the stabilizing effect due to increased heterozygosity and the disruptive effect caused by breakdown of genomic co-adaptation. Here we have compared the amount of fluctuating asymmetry across a transect of the hybrid zone between the two European subspecies of the house mouse (Mus musculus domesticus and M. m. musculus) in Denmark. For the first time in any natural hybrid zone we found an increased developmental stability in the populations with mixed genomes. Moreover, the apparently beneficial effect of hybridization on the developmental stability of the hybrid mice contrasts with the results of both genetic and parasitological studies which show that hybrid dysgenesis occurs in this zone. Our results suggest that the barrier to gene flow in the Mus musculus hybrid zone may result from the disruption of relatively few gene systems. They also lead us to reassess the relation between developmental stability expressed as fluctuating asymmetry, co-adaptation and overall fitness.