Pesticide exposure affects DNA methylation patterns in natural populations of a mayfly.

Chemical pollutants derived from agricultural activities represent a major threat to freshwater biota. Despite growing evidence involving epigenetic processes, such as DNA methylation, in response to pesticide contamination in agroecosystems, research on wild populations of non-model species remains scarce, particularly for endemic freshwater arthropods. Using the MethylRAD method, this study investigates whether exposure to pesticide contamination in natural populations of the endemic mayfly A. torrens produces genome wide changes in levels of DNA methylation. From a total of 1,377,147 MethylRAD markers produced from 285 specimens collected at 30 different study sites along the Limarí watershed of north-central Chile, six showed significant differential methylation between populations exposed and unexposed to pesticides. In all cases the effect of pesticides was positive, independent and stronger than the effects detected for other spatial and environmental factors. Only one candidate marker appeared correlated significantly with additional variables, nitrate and calcium levels, which also reflects the impact of agrichemicals and could additionally suggest, to a lower extent, antagonistic effects of mineral salts concentration for this specific marker. These results suggest that the effect of pesticide exposure on methylation levels is apparent at these six MethylRAD markers in A. torrens populations. Such data is challenging to obtain in natural populations and is, for the most part, lacking in ecotoxicological studies. Our study shows that DNA methylation processes are involved in the response to pesticide contamination in populations of the mayfly A. torrens in their natural habitat, and provides new evidence regarding the impact of pesticide contamination and agricultural activities on the endemic fauna of lotic ecosystems.

Spatial autocorrelation signatures of ecological determinants on plant community characteristics in high Andean wetlands.

Understanding how biological communities are shaped is a central tenet of community ecology. Recent evidence highlights the potential of decoupling diversity spatial autocorrelation into its positive and negative components to reveal community assembly processes that would otherwise remain undetected, as well as to improve understanding of their impacts on different facets of diversity. Yet, such approaches have only been implemented to investigate the effects of a few assembly drivers on a small number of diversity components. Here, we used high Andean wetland plant communities over a strong latitudinal gradient to investigate the effects of various ecological factors on spatial autocorrelation patterns of nine community metrics with different informative values, including measures of richness, dominance, evenness and beta-diversity. By combining Moran's Eigenvector Maps, partial least squares structural equation modeling, and regression analyses, we revealed two groups of community parameters presenting contrasting spatial patterns due to specific sensitivities to ecological factors. While environmental variation and wetland connectivity increased positive spatial autocorrelation in richness and dominance-related parameters, species co-occurrence promoted negative spatial autocorrelation in evenness-related parameters. These results offer new insights regarding both how ecological processes affect species assembly, as well as the information captured by classical taxonomic parameters.

Genome-wide genetic diversity yields insights into genomic responses of candidate climate-selected loci in an Andean wetland plant.

Assessing population evolutionary potential has become a central tenet of conservation biology. Since adaptive responses require allelic variation at functional genes, consensus has grown that genetic variation at genes under selection is a better surrogate for adaptive evolutionary potential than neutral genetic diversity. Although consistent with prevailing theory, this argument lacks empirical support and ignores recent theoretical advances questioning the very concept of neutral genetic diversity. In this study, we quantified genome-wide responses of single nucleotide polymorphism loci linked to climatic factors over a strong latitudinal gradient in natural populations of the high Andean wetland plant, Carex gayana, and then assessed whether genetic variation of candidate climate-selected loci better predicted their genome-wide responses than genetic variation of non-candidate loci. Contrary to this expectation, genomic responses of climate-linked loci only related significantly to environmental variables and genetic diversity of non-candidate loci. The effects of genome-wide genetic diversity detected in this study may be a result of either the combined influence of small effect variants or neutral and demographic factors altering the adaptive evolutionary potential of C. gayana populations. Regardless of the processes involved, our results redeem genome-wide genetic diversity as a potentially useful indicator of population adaptive evolutionary potential.

Developmental instability is associated with estrogenic endocrine disruption in the Chilean native fish species, Trichomycterus areolatus.

Endocrine disrupting compounds (EDCs) are widespread contaminants that alter the normal functioning of the endocrine system. While they cause dysfunctions in essential biological processes, it is unclear whether EDCs also impact developmental stability. In the present study, we investigated the occurrence of estrogenic endocrine disrupting compounds in a small watershed of south-central Chile impacted by anthropogenic activities. Then, we assessed their relationship with internal levels of estrogenic active compounds and fluctuating asymmetry (FA), a proxy of developmental stability in organisms with bilateral symmetry, in a native fish species (Trichomycterus areolatus). Yeast estrogenic screen assays were performed to measure estrogenic activity in river sediments and in male fish tissues collected from 17 sites along the Chillán watershed, and geometric morphometrics used to estimate fluctuating asymmetry based on the shapes of 248 fish skulls. Estrogenic activity was detected both in sediments and male fish tissues at concentrations of up to 1005 ng and 83 ng 17ß-estradiol equivalent/kg dw, respectively. No significant correlation was found between the two. However, fish tissue estrogenicity, water temperature and dissolved oxygen explained >80% of the FA population variation. By showing a significant relationship between estrogenic activity and FA of T. areolatus, our results indicate that developmental stability can be altered by estrogenic endocrine disruption, and that FA can be a useful indicator of sub-lethal stress in T. areolatus populations.

Assessment of Gene Expression Biomarkers in the Chilean Pencil Catfish, Trichomycterus areolatus, from the Choapa River Basin, Coquimbo Chile.

The objective of this study was to describe changes in the gene expression in the Chilean catfish, Trichomycterus areolatus, based on their geographic location within the Choapa River. Genes of choice included those that are biomarkers of exposure to metals, oxidative stress, and endocrine disruption. Male and female T. areolatus were sampled from four sites in January 2015 differently impacted by human activities. In males, but not females, hepatic gene expression of heat shock protein (HSP70) and cytochrome P450 1A (CYP1A) were significantly elevated at the site adjacent to the small city of Salamanca, relative to the other sites. In females, hepatic HSP70, the aryl hydrocarbon receptor (AHR), and the estrogen responsive genes, vitellogenin (VTG) and estrogen receptor alpha (ERα), were significantly lower at the site located furthest downstream. A similar downstream pattern of lower expression levels also was found in ovarian tissue for the genes, HSP70 and ERα. Gill gene expression showed a unique pattern in females as levels of metallothionein were elevated at the site furthest downstream. While analytical chemistry of water samples provided limited evidence of agrichemical contamination, the gene expression data are consistent with an exposure to agrichemicals and metals. T. areolatus may be a valuable sentinel organism and its use as a bioindicator species in some rivers within Chile can provide considerable insight, particularly in situations analytical chemistry is limited by environmental constraints.

Pesticide contamination drives adaptive genetic variation in the endemic mayfly Andesiops torrens within a semi-arid agricultural watershed of Chile.

Agrichemical contamination can provoke evolutionary responses in freshwater populations. It is a particularly relevant issue in semi-arid regions due to the sensitivity of endemic species to pollutants and to interactions with temperature stress. This paper investigates the presence of pesticides in rivers within a semi-arid agricultural watershed of Chile, testing for their effects on population genetic characteristics of the endemic mayfly Andesiops torrens (Insecta, Ephemeroptera). Pesticides were detected in sediment samples in ten out of the 30 sites analyzed throughout the upper part of the Limarí watershed. To study the evolutionary impact of such contamination on A. torrens, we used a genome-wide approach and analyzed 2056 single nucleotide polymorphisms (SNPs) loci in 551 individuals from all sites. Genetic differentiation was weak between populations, suggesting high gene flow across the study area. While we did not find evidence of pesticide effects on genetic diversity nor on population differentiation, the allele frequency of three outlier SNP loci correlated significantly with pesticide occurrence. Interrogation of genomic resources indicates that two of these SNPs are located within functional genes that encode for the low-density lipoprotein receptor-related protein 2 and Dumpy, both potentially involved in insect cuticle resistance processes. Such genomic signatures of local adaptation are indicative of past adverse effects of pesticide exposure on the locally adapted populations. Our results reveal that A. torrens is sensitive to pesticide exposure, but that a high gene flow may confer resilience to contamination. This research supports the contention that A. torrens is an ideal model organism to study evolutionary responses induced by pesticides on non-target, endemic species.

Landscape connectivity among remnant populations of guanaco (Lama guanicoe Müller, 1776) in an arid region of Chile impacted by global change.

Connectivity between populations plays a key role in the long-term persistence of species in fragmented habitats. This is of particular concern for biodiversity preservation in drylands, since water limited landscapes are typically characterized by little suitable habitat cover, high habitat fragmentation, harsh matrices, and are being rapidly degraded at a global scale. In this study, we modelled landscape connectivity between 11 guanaco Lama guanicoe populations in Chile's arid Norte Chico, a region that supports the last remnant coastal populations of this emblematic herbivore indigenous to South America. We produced a habitat suitability model to derive a regional surface resistance map, and used circuit theory to map functional connectivity, investigate the relative isolation between populations, and identify those that contribute most to the patch connectivity network. Predicted suitable habitat for L. guanicoe represented about 25% of the study region (i.e., 29,173 km2) and was heterogeneously distributed along a continuous stretch along the Andes, and discontinuous patches along the coast. As a result, we found that high connectivity current flows in the mid and high Andes formed a wide, continuous connectivity corridor, enabling connectivity between all high Andean populations. Coastal populations, in contrast, were more isolated. These groups demonstrate no inter-population connectivity between themselves, only with higher altitude populations, and for two of them, animal movement was linked to the effectiveness of wildlife crossings along the Pan-American highway. Our results indicate that functional connectivity is an issue of concern for L. guanicoe in Chile's Norte Chico, implying that future conservation and management plans should emphasize strategies aimed at conserving functional connectivity between coastal and Andean populations, as well as the protection of habitat patches likely to act as stepping stones within the connectivity network.

Partitioning genetic and species diversity refines our understanding of species-genetic diversity relationships.

Disentangling the origin of species-genetic diversity correlations (SGDCs) is a challenging task that provides insight into the way that neutral and adaptive processes influence diversity at multiple levels. Genetic and species diversity are comprised by components that respond differently to the same ecological processes. Thus, it can be useful to partition species and genetic diversity into their different components to infer the mechanisms behind SGDCs. In this study, we applied such an approach using a high-elevation Andean wetland system, where previous evidence identified neutral processes as major determinants of the strong and positive covariation between plant species richness and AFLP genetic diversity of the common sedge Carex gayana. To tease apart putative neutral and non-neutral genetic variation of C. gayana, we identified loci putatively under selection from a dataset of 1,709 SNPs produced using restriction site-associated DNA sequencing (RAD-seq). Significant and positive relationships between local estimates of genetic and species diversities (α-SGDCs) were only found with the putatively neutral loci datasets and with species richness, confirming that neutral processes were primarily driving the correlations and that the involved processes differentially influenced local species diversity components (i.e., richness and evenness). In contrast, SGDCs based on genetic and community dissimilarities (ß-SGDCs) were only significant with the putative non-neutral datasets. This suggests that selective processes influencing C. gayana genetic diversity were involved in the detected correlations. Together, our results demonstrate that analyzing distinct components of genetic and species diversity simultaneously is useful to determine the mechanisms behind species-genetic diversity relationships.

Genetic variation of loci potentially under selection confounds species-genetic diversity correlations in a fragmented habitat.

Positive species-genetic diversity correlations (SGDCs) are often thought to result from the parallel influence of neutral processes on genetic and species diversity. Yet, confounding effects of non-neutral mechanisms have not been explored. Here, we investigate the impact of non-neutral genetic diversity on SGDCs in high Andean wetlands. We compare correlations between plant species diversity and genetic diversity (GD) calculated with and without loci potentially under selection (outlier loci). The study system includes 2188 specimens from five species (three common aquatic macroinvertebrate and two dominant plant species) that were genotyped for 396 amplified fragment length polymorphism loci. We also appraise the importance of neutral processes on SGDCs by investigating the influence of habitat fragmentation features. Significant positive SGDCs were detected for all five species (mean SGDC = 0.52 ± 0.05). While only a few outlier loci were detected in each species, they resulted in significant decreases in GD and in SGDCs. This supports the hypothesis that neutral processes drive species-genetic diversity relationships in high Andean wetlands. Unexpectedly, the effects on genetic diversity GD of the habitat fragmentation characteristics in this study increased with the presence of outlier loci in two species. Overall, our results reveal pitfalls in using habitat features to infer processes driving SGDCs and show that a few loci potentially under selection are enough to cause a significant downward bias in SGDC. Investigating confounding effects of outlier loci thus represents a useful approach to evidence the contribution of neutral processes on species-genetic diversity relationships.

The role of spatial processes and environmental determinants in microgeographic shell variation of the freshwater snail Chilina dombeyana (Bruguière, 1789).

Wildlife data often show spatial organization, demonstrating positive correlations either as a result of processes occurring over the landscape or due to the influence of spatially structured environmental variables. It is, thus, essential to consider non-random spatial structure when evaluating the underlying causes of biological variation. In this study, we analyzed the population structure of Chilina dombeyana shell morphology of 14 populations that are close geographically and belong to the same hydrographic basin. We utilized a variation partitioning approach to evaluate the importance of spatial processes, such as migration, acting over the landscape, and environmental characteristics, including habitat and hydrologic characteristics, and the occurrence of aquatic predators in promoting between population variation. Our results demonstrate spatially structured variation in C. dombeyana shell morphology, with populations living near each other having more similar shell sizes than populations living farther apart. The shell size variation partition indicated that both spatially structured environmental factors and genetic relationships resulting from migration or shared common ancestry may explain this pattern. Shell shape variation, in contrast, was found to be essentially under the influence of non-spatially structured environmental factors, with habitat and water characteristics accounting for about half of the total variation among populations. The large proportion of the variation in shell size that is spatially structured demonstrates that spatial structure on morphological traits might be strong and highlights the need to consider such phenomenon in intraspecific studies of phenotypic evolution.

Permanent genetic resources added to Molecular Ecology Resources Database 1 February 2011-31 March 2011.

This article documents the addition of 111 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Acipenser oxyrinchus desotoi, Anopheles nuneztovari sensu lato, Asellus aquaticus, Calopteryx splendens, Calopteryx virgo, Centaurea aspera, Centaurea seridis, Chilina dombeyana, Proctoeces cf. lintoni and Pyrenophora teres f. teres.

Estrogen pollution in a highly productive ecosystem off central-south Chile.

While the presence of steroid estrogens in the environment has become a major environmental and health concern, their occurrence in coastal sediments remains poorly characterized. In this study, we measured the levels of three natural (estrone, 17ß-estradiol, estriol) and one synthetic (17α-ethinylestradiol) estrogens in 54 coastal sediment samples collected from nine locations off central-southern Chile. Steroid estrogens were found in every sample. Remarkably high levels of 17α-ethinylestradiol were detected, reaching up to 48.14 ng/g dry weight. As a result, the global estrogenic loads were estimated to be high at all sites. Interestingly, they were found to correlate with the size of human populations served by sewage plants. Our study indicates that 17α-ethinylestradiol may accumulate in coastal sediments. The possible impact of this highly potent synthetic estrogen on the biota of the marine ecosystem off central-south Chile and on human health remains an open question.

A theoretical estimation of the concentration of steroid estrogens in effluents released from municipal sewage treatment plants into aquatic ecosystems of central-southern Chile.

Endocrine disorders associated with sewage effluents have been documented in aquatic species from various regions of the world and sewage treatment works (STWs) are now widely recognized as one of the major discharge source of endocrine disrupting compounds. Steroid estrogens usually emerge as the main contributors to the endocrine disrupting capacity of municipal sewage effluents. Because human wastes are believed to be the primary source of release of steroid estrogens in watercourses, the presence of these compounds in aquatic systems is likely to constitute a pervasive ecological problem. In spite of that, the endocrine disrupting impact of sewage effluents has rarely been investigated in South America. In this paper, we used Johnson and Williams' predictive model to estimate the concentration of steroid estrogens in effluents released from 38 municipal STWs of central-southern Chile and to assess steroid estrogen concentrations in rivers. In STW effluents, we estimated the estrogen concentrations to range from 9.35 to 739.92 ng/L for estrone, 1.03 to 81.74 ng/L for estradiol and 0.38 to 30.56 ng/L for ethynylestradiol. Overall, the predicted estrogen concentrations are significantly higher than those reported for STW effluents in the literature. This can be explained by demographic and sewage flow differences between Chile and industrialized western countries. Predicted steroid estrogen concentrations at river sites indicate that endocrine disruption in fish is likely to occur in the Itata catchment. However, future research is needed to attest this and to evaluate the real impact of the STW discharges into central-southern Chile's marine and freshwater environments.

Quantitative trait loci for peripheral blood cell counts: a study in baboons.

Increasingly, baseline peripheral blood cell counts are implicated as risk factors for common complex diseases. While genetic influences on these hematologic parameters are firmly established, the genetic architecture of the blood counts is still poorly understood. In this article we used data from 582 healthy pedigreed baboons and variance components methods to localize quantitative trait loci (QTLs) influencing complete blood count variables. Besides performing genome-wide linkage scans for each trait individually, we conducted bivariate linkage analyses for all pairwise trait combinations to also identify pleiotropic QTLs influencing several blood counts. While significant and suggestive QTLs were localized throughout the genome (LOD range: 1.5-3.5), chromosomal regions associated with the expression of various hematologic parameters stand out. In particular, our results provide significant and consistent evidence for a QTL on the orthologous human chromosome 1p that is shared by several blood counts, mainly erythrocyte parameters. In addition, multiple suggestive evidence of linkage was detected on the orthologous human chromosomes 10 (near the q-terminus) and 19 (centromeric section). Future studies should help identify the genes responsible for these QTL and elucidate their role on baseline variation in hematologic indicators of health and disease.

The form of sexual selection on male genitalia cannot be inferred from within-population variance and allometry - a case study in Aquarius remigis.

Male genital morphology in insects and arachnids is characterized by static hypoallometry and low intrapopulational levels of phenotypic variation relative to other male traits. The one-size-fits-all model of genital evolution attributes these patterns to stabilizing sexual selection. This model relies on the assumption that the observed patterns of variation and allometry reflect the form of sexual selection acting these traits. We test this by examining the patterns of scaling and trait variation for a set of genitalic and somatic morphological traits in male water striders (Aquarius remigis). This suite of traits is of particular interest because previous work has shown that the genitalic traits are under strong directional selection whereas the somatic traits are under either weak directional or stabilizing selection. Because the selection regime for these traits is known, we can, for the first time, test the purported relationship between trait variation, scaling, and the form of sexual selection. We show that the patterns of variation and scaling of these traits differ sharply from those predicted for traits experiencing strong directional sexual selection. Specifically, the male genital structures show static hypoallometry and low intrapopulational levels of phenotypic variation relative to other male traits, in spite of consistent, strong, directional sexual selection. These scaling relationships and levels of variation are typical of genital traits in other insect species, where they have been presumed to reflect stabilizing sexual selection. Our data clearly refute the assumption of the one-size-fits-all hypothesis that hypoallometric scaling of genitalic traits implies stabilizing selection. We discuss the implications of this finding and propose future directions for improving our current understanding of genital evolution in arthropods.