Restoring groundwater quality at the drinking water catchment scale: A multidisciplinary and participatory approach.

Preserving or restoring the quality of groundwater resources with regard to nitrate is a major challenge. To date, useful and easily applicable tools to identify the best measures to implement at local scale are lacking. An innovative methodology is proposed to identify cost-effective restoration measures at the drinking water catchment scale. The methodology is based on the articulation of two tools: a model simulating nitrate groundwater contamination time series and an economic evaluation, within a participatory approach. It was applied to a representative drinking water catchment in Northwest France that has been affected by nitrate contamination for decades. Five scenarios of measures (changes in fertilization and intercropping practices, or in cropping patterns) were co-constructed with stakeholders, evaluated in terms of cost and impact on groundwater nitrate concentration, and discussed with stakeholders. Overall, two scenarios stand out. Introducing hemp in crop rotations is the only scenario generating an economic benefit, but for a very low impact on nitrate concentration (decrease of 4 mg/l by 2050 for 10% of hemp in crop rotations). Introducing alfalfa in crop rotations is the most effective measure to decrease nitrate concentration (decrease of 23 mg/l by 2050 for 20% of alfalfa in crop rotations), and for a moderate cost compared to the other scenarios (25-51 €/ha/year). Results show that substantial changes in cropping patterns - more important than those imagined initially by stakeholders (e.g. converting 40% of cropland to alfalfa) are needed to restore groundwater quality targets in the medium-long term. Measures deemed promising and achievable by the stakeholders proved to be insufficiently effective. The approach developed has been shown to provide valued and trusted information to stakeholders and to objectify debates. Stakeholders have shown interest in the evaluation of costs and further socio-economic information, in addition to the evaluation of the effectiveness of measures on groundwater quality, validating the multidisciplinary dimension of the approach. The approach has the advantage of being easy to implement, and is therefore applicable to other study sites where needed.

Triple-Element Compound-Specific Stable Isotope Analysis (3D-CSIA): Added Value of Cl Isotope Ratios to Assess Herbicide Degradation.

Multielement isotope fractionation studies to assess pollutant transformation are well-established for point-source pollution but are only emerging for diffuse pollution by micropollutants like pesticides. Specifically, chlorine isotope fractionation is hardly explored but promising, because many pesticides contain only few chlorine atoms so that "undiluted" position-specific Cl isotope effects can be expected in compound-average data. This study explored combined Cl, N, and C isotope fractionation to sensitively detect biotic and abiotic transformation of the widespread herbicides and groundwater contaminants acetochlor, metolachlor, and atrazine. For chloroacetanilides, abiotic hydrolysis pathways studied under acidic, neutral, and alkaline conditions as well as biodegradation in two soils resulted in pronounced Cl isotope fractionation (εCl from -5.0 ± 2.3 to -6.5 ± 0.7‰). The characteristic dual C-Cl isotope fractionation patterns (ΛC-Cl from 0.39 ± 0.15 to 0.67 ± 0.08) reveal that Cl isotope analysis provides a robust indicator of chloroacetanilide degradation. For atrazine, distinct ΛC-Cl values were observed for abiotic hydrolysis (7.4 ± 1.9) compared to previous reports for biotic hydrolysis and oxidative dealkylation (1.7 ± 0.9 and 0.6 ± 0.1, respectively). The 3D isotope approach allowed differentiating transformations that would not be distinguishable based on C and N isotope data alone. This first data set on Cl isotope fractionation in chloroacetanilides, together with new data in atrazine degradation, highlights the potential of using compound-specific chlorine isotope analysis for studying in situ pesticide degradation.

Applying gene regulatory network logic to the evolution of social behavior.

Animal behavior is ultimately the product of gene regulatory networks (GRNs) for brain development and neural networks for brain function. The GRN approach has advanced the fields of genomics and development, and we identify organizational similarities between networks of genes that build the brain and networks of neurons that encode brain function. In this perspective, we engage the analogy between developmental networks and neural networks, exploring the advantages of using GRN logic to study behavior. Applying the GRN approach to the brain and behavior provides a quantitative and manipulative framework for discovery. We illustrate features of this framework using the example of social behavior and the neural circuitry of aggression.

Influence of hydrodynamics on the water pathway and spatial distribution of pesticide and metabolite concentrations in constructed wetlands.

Constructed wetlands (CWs) are likely to reduce pesticide levels reaching surface water. However, the distribution of the water flow path between the main channel and isolated areas may influence global pesticide mitigation. Little information is known about the influence of water pathways on pesticide mitigation. Thus, we performed tracer experiments at low and high flow rates (0.5 L/s and 4-7 L/s) in a pond CW and ditch CW to determine the localization of various hydraulic zones and to understand their implication on pesticide mitigation. The hydraulic performance reflecting the fraction of water transported from inlet to outlet passing through the whole of CW, was greater for the pond CW than for the ditch CW regardless of the flow rate, and greater at mean flow rates (MF) than at low flow rates (LF) due to a lower proportion of isolated areas at a MF (11%-68%) than at LF (38%-89%). Dispersion governed the water transport inside the isolated areas and the water convection inside the main channel. Consequently, dissolved pesticide concentrations are heterogeneously distributed in the CWs, i.e., in the main channel and isolated area, for both flow rates. However, one month after a no-flow period, this heterogeneity disappears, and dissolved pesticide concentrations become similar in the water of the whole CW due to dispersion. Furthermore, sedimentation and storage in sediments were greater in the isolated area than in the main channel, which is possibly due to a lower speed flow rate and a higher hydraulic residence time (HRT) in the isolated area than in the main channel. Thus, isolated areas act as effective's zones to mitigate pesticides from dissolved and particulate phases inside the CW during a complete drainage season (i.e., succession of high/low/no-flow periods).

Reductionist thinking and animal models in neuropsychiatric research.

Reductionist thinking in neuroscience is manifest in the widespread use of animal models of neuropsychiatric disorders. Broader investigations of diverse behaviors in non-model organisms and longer-term study of the mechanisms of plasticity will yield fundamental insights into the neurobiological, developmental, genetic, and environmental factors contributing to the "massively multifactorial system networks" which go awry in mental disorders.

Early life manipulations of vasopressin-family peptides alter vocal learning.

Vocal learning from social partners is crucial for the successful development of communication in a wide range of species. Social interactions organize attention and enhance motivation to learn species-typical behaviour. However, the neurobiological mechanisms connecting social motivation and vocal learning are unknown. Using zebra finches (Taeniopygia guttata), a ubiquitous model for vocal learning, we show that manipulations of nonapeptide hormones in the vasopressin family (arginine vasotocin, AVT) early in development can promote or disrupt both song and social motivation. Young male zebra finches, like human infants, are socially gregarious and require interactive feedback from adult tutors to learn mature vocal forms. To investigate the role of social motivational mechanisms in song learning, in two studies, we injected hatchling males with AVT or Manning compound (MC, a nonapeptide receptor antagonist) on days 2-8 post-hatching and recorded song at maturity. In both studies, MC males produced a worse match to tutor song than controls. In study 2, which experimentally controlled for tutor and genetic factors, AVT males also learned song significantly better compared with controls. Furthermore, song similarity correlated with several measures of social motivation throughout development. These findings provide the first evidence that nonapeptides are critical to the development of vocal learning.

Developmental effects of vasotocin and nonapeptide receptors on early social attachment and affiliative behavior in the zebra finch.

Zebra finches demonstrate selective affiliation between juvenile offspring and parents, which, like affiliation between pair partners, is characterized by proximity, vocal communication and contact behaviors. This experiment tested the hypothesis that the nonapeptide arginine vasotocin (AVT, avian homologue of vasopressin) and nonapeptide receptors play a role prior to fledging in the development of affiliative behavior. Zebra finch hatchlings of both sexes received daily intracranial injections (post-hatch days 2-8) of either AVT, Manning Compound (MC, a potent V1a receptor antagonist) or saline (vehicle control). The social development of both sexes was assessed by measuring responsiveness to isolation from the family and subsequent reunion with the male parent after fledging. In addition, we assessed the changes in affiliation with the parents, unfamiliar males, and unfamiliar females each week throughout juvenile development. Compared to controls, MC subjects showed decreased attachment to the parents and MC males did not show the normal increase in affiliative interest in opposite sex individuals as they reached reproductive maturity. In contrast, AVT subjects showed a sustained affiliative interest in parents throughout development, and males showed increased interest in opposite sex conspecifics as they matured. These results provide the first evidence suggesting that AVT and nonapeptide receptors play organizational roles in social development in a bird.

Coevolution of Parental Care, Parasitic, and Resistance Efforts in Facultative Parasitism.

Here we develop a tug-of-war game theory model of mixed-strategy facultative parasitism. In this framework, individuals decide how to strategically invest in parental care effort, parasitism, and resistance to being parasitized, choosing their investments to respond optimally to their opponent's behavior and vice versa. We have implemented the model in the well-studied case of mixed-strategy conspecific brood parasitism, which occurs when a female raises her own clutch of eggs and flexibly lays some eggs in a conspecific's nest. However, the nest of a parasitic female in this case may also be parasitized. We model this as a tug-of-war, allowing parasitic and resistance efforts to strategically coevolve with each other. We then derive expressions for parasitism outcomes commonly observed in the field. The model also captures the trade-offs between parental care and parasitism when both are possible. We make a number of novel predictions about the rate of successful versus attempted parasitism as well as how parental care effort, resistance to parasitism, and fitness vary as functions of group size and individual differences between players in energy budget and production efficiency. Although we focus primarily on conspecific brood parasitism, the model is general enough to be extended to other systems in which facultative parasitism is possible, including the parasitism of food, fertilizations, nest sites, or other resources from conspecifics.

Advantages of online SPE coupled with UPLC/MS/MS for determining the fate of pesticides and pharmaceutical compounds.

Laboratory experimentation is essential for our understanding of the fate and behaviour of pollutants. Many analytical techniques exist, but they all have disadvantages either in terms of sensitivity or of selectivity. The number of samples that can be analysed, the low volume of samples available during the experiment and the need to identify different degradates are all obstacles that new techniques are able to overcome. The work presented here summarizes progress in the field of metrology as concerns online solid phase extraction technology coupled with liquid chromatography followed by tandem mass spectrometry detection. Recently developed analytical techniques were validated for both 18 pesticides and their degradates and 17 pharmaceuticals and their degradates. Limits of quantification from 20 to 70 ng L(-1) for pharmaceuticals and from 15 to 25 ng L(-1) for pesticides and metabolites have been obtained, with linearity range up to 1 µg L(-1). The limits of quantification of a few nanograms per litre, the possibility of working on less than 1 mL of sample and the simultaneous quantification of the target products and their transformation products are all advantages that are demonstrated by two environmental applications. The first application concerns the evaluation of ecotoxicological effects of pesticides on aquatic organisms exposed in mesocosms. The second application aims to determine the adsorption constants of pharmaceutical molecules on soils and river sediments. For both applications, the robustness, range of linearity and limit of quantification of the developed analytical methods satisfy the requirements for laboratory experiments conducted under controlled conditions. Specific constraints generated by this type of experiment (adding CaCl2 for the adsorption study and filtration of the water coming from the mesocosms) were not found to limit the use of online SPE. These two preliminary studies show that new experimental fields are possible thanks to online solid phase extraction coupled with liquid chromatography.

Low concentrations of ethylene bisdithiocarbamate pesticides maneb and mancozeb impair manganese and zinc homeostasis to induce oxidative stress and caspase-dependent apoptosis in human hepatocytes.

The worldwide and intensive use of phytosanitary compounds results in environmental and food contamination by chemical residues. Human exposure to multiple pesticide residues is a major health issue. Considering that the liver is not only the main organ for metabolizing pesticides but also a major target of toxicities induced by xenobiotics, we studied the effects of a mixture of 7 pesticides (chlorpyrifos-ethyl, dimethoate, diazinon, iprodione, imazalil, maneb, mancozeb) often detected in food samples. Effects of the mixture was investigated using metabolically competent HepaRG cells and human hepatocytes in primary culture. We report the strong cytotoxicity of the pesticide mixture towards hepatocytes-like HepaRG cells and human hepatocytes upon acute and chronic exposures at low concentrations extrapolated from the Acceptable Daily Intake (ADI) of each compound. Unexpectedly, we demonstrated that the manganese (Mn)-containing dithiocarbamates (DTCs) maneb and mancozeb were solely responsible for the cytotoxicity induced by the mixture. The mechanism of cell death involved the induction of oxidative stress, which led to cell death by intrinsic apoptosis involving caspases 3 and 9. Importantly, this cytotoxic effect was found only in cells metabolizing these pesticides. Herein, we unveil a novel mechanism of toxicity of the Mn-containing DTCs maneb and mancozeb through their metabolization in hepatocytes generating the main metabolite ethylene thiourea (ETU) and the release of Mn leading to intracellular Mn overload and depletion in zinc (Zn). Alteration of the Mn and Zn homeostasis provokes the oxidative stress and the induction of apoptosis, which can be prevented by Zn supplementation. Our data demonstrate the hepatotoxicity of Mn-containing fungicides at very low doses and unveil their adverse effect in disrupting Mn and Zn homeostasis and triggering oxidative stress in human hepatocytes.

Carbon isotope ratio measurements of glyphosate and AMPA by liquid chromatography coupled to isotope ratio mass spectrometry.

The interest in compound-specific isotope analysis for product authenticity control and source differentiation in environmental sciences has grown rapidly during the last decade. However, the isotopic analysis of very polar analytes is a challenging task due to the lack of suitable chromatographic separation techniques which can be used coupled to isotope ratio mass spectrometry. In this work, we present the first method to measure carbon isotope compositions of the widely applied herbicide glyphosate and its metabolite aminomethylphosphonic acid (AMPA) by liquid chromatography coupled to isotope ratio mass spectrometry. We demonstrate that this analysis can be carried out either in cation exchange or in reversed-phase separation modes. The reversed-phase separation yields a better performance in terms of resolution compared with the cation exchange method. The measurement of commercial glyphosate herbicide samples show its principal applicability and reveals a wide range of δ(13)C values between -24 and -34 ‰ for different manufacturers. The absolute minimum amounts required to perform a precise and accurate determination of carbon isotope compositions of glyphosate and AMPA were in the sub-microgram range. The method proposed is sensitive enough to further perform the experiments that are necessary to better understand the carbon isotope fractionation associated to the natural degradation of glyphosate into AMPA. Furthermore, it can be used for contaminant source allocation and product authenticity as well.

Oxytocin receptor antagonism during early vocal learning reduces song preference and imitation in zebra finches.

In species with vocal learning, acquiring species-typical vocalizations relies on early social orienting. In songbirds, for example, learning song requires dynamic social interactions with a "tutor" during an early sensitive period. Here, we hypothesized that the attentional and motivational processes that support song learning recruit the oxytocin system, which is well-understood to play a role in social orienting in other species. Juvenile male zebra finches naïve to song were each tutored by two unfamiliar adult males. Before exposure to one tutor, juveniles were injected subcutaneously with oxytocin receptor antagonist (OTA; ornithine vasotocin) and before exposure to the other, saline (control). Treatment with OTA reduced behaviors associated with approach and attention during tutoring sessions. Using a novel operant paradigm to measure preference while balancing exposure to the two tutor songs, we showed that the juveniles preferred to hear the song of the control tutor. Their adult songs more closely resembled the control tutor's song, and the magnitude of this difference was predicted by early preference for control over OTA song. Overall, oxytocin antagonism during exposure to a tutor seemed to bias juveniles against that tutor and his song. Our results suggest that oxytocin receptors are important for socially-guided vocal learning.

Pesticides and their metabolites in European groundwater: Comparing regulations and approaches to monitoring in France, Denmark, England and Switzerland.

Pesticides, i.e. plant protection products (PPP), biocides and their metabolites, pose a serious threat to groundwater quality and groundwater dependent ecosystems. Across large parts of Europe these compounds are monitored in groundwater to ensure compliance with the European Water Framework Directive (WFD), the Groundwater Directive (GWD) and Drinking water Directive (DWD). European regulation concerning the placing of PPP on the market includes groundwater monitoring as a higher tier of the regulatory procedure. Nevertheless, the lists of compounds to be monitored vary from one directive to another and between countries. The implementation of monitoring strategies for these directives and other national drivers, differs across Europe. This is illustrated using case studies from France, Denmark (EU member states), England (part of the EU up to January 2020) and Switzerland (associated country). The collection of data (e.g. monitoring design and analytical approaches) and dissemination at national and European level and the scale of data reporting to EU is country-specific. Data generated by the implementation of WFD and DWD can be used for retrospective purposes in the context of PPP registration whereas the post-registration monitoring data generated by the product applicants are generally only directly available to the regulators. This lack of consistency and strategic coordination between thematic regulations is partly compensated by national regulations. This paper illustrates the benefits of a common framework for regulation in Europe but shows that divergent national approaches to monitoring and reporting on pesticides in groundwater makes the task of assessment across Europe challenging.

Dynamic molecular evolution of a supergene with suppressed recombination in white-throated sparrows.

In white-throated sparrows, two alternative morphs differing in plumage and behavior segregate with a large chromosomal rearrangement. As with sex chromosomes such as the mammalian Y, the rearranged version of chromosome two (ZAL2m) is in a near-constant state of heterozygosity, offering opportunities to investigate both degenerative and selective processes during the early evolutionary stages of 'supergenes.' Here, we generated, synthesized, and analyzed extensive genome-scale data to better understand the forces shaping the evolution of the ZAL2 and ZAL2m chromosomes in this species. We found that features of ZAL2m are consistent with substantially reduced recombination and low levels of degeneration. We also found evidence that selective sweeps took place both on ZAL2m and its standard counterpart, ZAL2, after the rearrangement event. Signatures of positive selection were associated with allelic bias in gene expression, suggesting that antagonistic selection has operated on gene regulation. Finally, we discovered a region exhibiting long-range haplotypes inside the rearrangement on ZAL2m. These haplotypes appear to have been maintained by balancing selection, retaining genetic diversity within the supergene. Together, our analyses illuminate mechanisms contributing to the evolution of a young chromosomal polymorphism, revealing complex selective processes acting concurrently with genetic degeneration to drive the evolution of supergenes.

Pesticides in groundwater at a national scale (France): Impact of regulations, molecular properties, uses, hydrogeology and climatic conditions.

Contaminants in groundwater are a major issue worldwide. Temporal trends of such occurrences in French groundwaters were evaluated for several active substances of pesticides belonging to different chemical classes, to identify key factors explaining groundwater contamination. Our study relied on exploitation of a French national database (ADES, created in the mid-1990s and remarkable for the available data, including over 88 million analyses). Temporal changes in the frequency of exceeding a reference value of 0.1 µg/L for several substances were determined at yearly and monthly scales. Such trends were examined by distinguishing different periods according to changes in regulations (new approval, withdrawal, or dose reduction), and were combined with data on effective rainfall as a proxy for groundwater recharge, on aquifer lithology, and on sales of active substances as a proxy for actual applications. A review of monthly data shows that a rapid transfer of pesticides with contrasting physico-chemical properties can occur after application in many aquifers, regardless of their lithology. For substances such as metolachlor, showing a sharp increase in sales, a clear relationship exists between quantities sold and frequency of exceeding the reference value. For other active substances, such as isoproturon or chlortoluron, frequencies of exceedance are governed by both sales and effective rainfall. Finally, the occurrence of active substances in groundwater several years after their withdrawal from the market is explained by at least three major mechanisms: the transfer time from soil into groundwater, processes of remobilization from soil and/or unsaturated zone, and no or low degradation in the saturated zone. While these processes are well documented for atrazine and different types of aquifers, they can be virtually unknown for other active substances.

Side Effects of Pesticides and Metabolites in Groundwater: Impact on Denitrification.

The impact of two pesticides (S-metolachlor and propiconazole) and their respective main metabolites (ESA-metolachlor and 1,2,4-triazole) on bacterial denitrification in groundwater was studied. For this, the denitrification activity and the bacterial diversity of a microbial community sampled from a nitrate-contaminated groundwater were monitored during 20 days in lab experiments in the presence or absence of pesticides or metabolites at 2 or 10 µg/L. The kinetics of nitrate reduction along with nitrite and N2O production all suggested that S-metolachlor had no or only little impact, whereas its metabolite ESA-metolachlor inhibited denitrification by 65% at 10 µg/L. Propiconazole and 1,2,4-triazole also inhibited denitrification at both concentrations, but to a lesser extent (29-38%) than ESA-metolachlor. When inhibition occurred, pesticides affected the reduction of nitrate into nitrite step. However, no significant differences were detected on the abundance of nitrate reductase narG and napA genes, suggesting an impact of pesticides/metabolites at the protein level rather than on denitrifying bacteria abundance. 16S rRNA gene Illumina sequencing indicated no major modification of bacterial diversity in the presence or absence of pesticides/metabolites, except for ESA-metolachlor and propiconazole at 10 µg/L that tended to increase or decrease Shannon and InvSimpson indices, respectively. General growth parameters suggested no impact of pesticides, except for propiconazole at 10 µg/L that partially inhibited acetate uptake and induced a decrease in microbial biomass. In conclusion, pesticides and metabolites can have side effects at environmental concentrations on microbial denitrification in groundwater and may thus affect ecosystem services based on microbial activities.

Elucidating heterogeneous nitrate contamination in a small basement aquifer. A multidisciplinary approach: NO3 isotopes, CFCs-SF6, microbiological activity, geophysics and hydrogeology.

Nitrate contamination of groundwater remains a major concern despite all the measures and efforts undertaken over the last decades to protect water resources. We focused on a small catchment in Brittany (France) facing nitrate pollution with concentrations over the European drinking water standard of 50 mg.L-1. This is a common situation in catchments where - supposedly effective - measures were applied for reducing the transfer of N to groundwater. At the scale of this small (~100 ha) basement aquifer, nitrate concentrations are very heterogeneous in the groundwater, sampled up to 15-20 m below the soil surface in several observation wells (hereafter referred as piezometers) and up to 110 m deep in a borehole drilled through a faulted area near the Spring (outlet of the catchment). We used complementary and robust approaches for exploring and constraining the driving parameters of nitrate transfer and distribution in groundwater. Detailed geological work and a geophysical electrical resistivity tomography survey identified the lithologies, tectonic structures and weathering layers. This highlighted a complex geological structure with several compartments delimited by faults, as well as the highly variable thickness of the weathered layer. It also illustrated the heterogeneity of the hydrosystem, some compartments appearing to be disconnected from the general groundwater flow. This was confirmed by geochemical analyses and by the mean apparent groundwater residence time based on CFCs-SF6 and noble-gas analyses, locally revealing old and nitrate-free groundwater, and very old water with a recharge temperature below than the current average temperature in the area, reflecting water dating back to the last period of glaciation (-19 to -17 ky). Nitrate isotopes clearly showed denitrification processes in a few piezometers, which was generally supported by microbiology and molecular biology results. This highlighted the presence of functional genes involved in denitrification as well as a capacity of the groundwater microbial community to denitrify when in situ conditions are favourable. This type of combined approach - covering chemistry, isotopic methods, dissolved gases, microbiological activity, geophysics and hydrogeology - appears to be indispensable for implementing the most relevant programme of measures and for accurately assessing their effectiveness, notably by considering the timeframe between implementation of the measures and their impact on groundwater quality.

Between- and within-sex variation in hormonal responses to psychological stress in a large sample of college students.

This study investigated (1) sex differences in hormonal responses to psychosocial stress; (2) the relation between variability in pre-test hormone concentrations and stress-induced hormonal changes; and (3) some possible sources of within-sex variation in pre-test hormone concentrations and in hormonal responses to the test in a large human subject population. To this end, changes in salivary concentrations of testosterone and cortisol in response to a mild psychosocial stressor (a set of computerized economic decision-making tests) were measured in a sample of over 500 MBA students. Males had higher concentrations of testosterone and cortisol than females both before and after the test. After taking effects of time of testing on hormone concentrations into account, testosterone showed a post-test decrease in males but not in females. Cortisol level increased in both sexes but the post-test increase was larger in females than in males. At the individual level, the pre-test concentrations of testosterone and cortisol predicted both the direction and the magnitude of the post-test hormone change, so that low pre-test hormone concentrations showed large post-test increases whereas high pre-test concentrations showed large post-test decreases. Within-sex variation in hormone concentrations was not accounted for by variation in 2D:4D digit length ratio, a marker of prenatal androgen exposure, but by social variables. Single males without a stable romantic partner had higher testosterone level than males with stable partners, and both males and females without a partner showed a greater cortisol response to the test than married individuals with or without children. Studies conducted with large sample sizes such as this one can help understand normative patterns of hormonal responses to psychosocial stimuli as well as identify the sources of interindividual variation in endocrine function.

Ecotype differences in aggression, neural activity and behaviorally relevant gene expression in cichlid fish.

In Lake Malawi, two ecologically distinct lineages of cichlid fishes (rock- vs sand-dwelling ecotypes, each comprised of over 200 species) evolved within the last million years. The rock-dwelling species (Mbuna) are aggressively territorial year-round and males court and spawn with females over rocky substrate. In contrast, males of sand-dwelling species are not territorial and instead aggregate on seasonal breeding leks in which males construct courtship "bowers" in the sand. However, little is known about how phenotypic variation in aggression is produced by the genome. In this study, we first quantify and compare behavior in seven cichlid species, demonstrating substantial ecotype and species differences in unconditioned mirror-elicited aggression. Second, we compare neural activity in mirror-elicited aggression in two representative species, Mchenga conophoros (sand-dwelling) and Petrotilapia chitimba (rock-dwelling). Finally, we compare gene expression patterns between these two species, specifically within neurons activated during mirror aggression. We identified a large number of genes showing differential expression in mirror-elicited aggression, as well as many genes that differ between ecotypes. These genes, which may underly species differences in behavior, include several neuropeptides, genes involved in the synthesis of steroid hormones and neurotransmitter activity. This work lays the foundation for future experiments using this emerging genetic model system to investigate the genomic basis of evolved species differences in both brain and behavior.

Water isotopes and chemical tools for understanding pesticide transfer in a watershed of the volcanic island of Martinique (French West Indies).

Monitoring of water quality over several years has revealed a persistent pesticide contamination of surface and groundwater in several Caribbean Islands, with pesticide concentrations locally over the drinking-water limit set in Europe, i.e. 0.1 µg L-1 per substance. For Martinique, mainly one pesticide, chlordecone (CLD), remains of major concern despite its withdrawal from the market in 1993. Since the first sampling campaign in 1999-2000, time and space variations of CLD concentrations in surface water and groundwater are still not well understood and difficult to correlate with climate, geological or hydrogeological contexts. We carried out a study in the Chalvet catchment (northeast Martinique) in order to understand more precisely how water movements may explain pesticide transfer. Various tools such as δ2H - δ18O and chemical parameters were used. Deuterium excess d was proven relevant for determining how CLD is transported in groundwater; it highlighted the role of the groundwater/surface water interaction in spatial and temporal variability of surface water quality. The resulting conceptual hydrogeological model also helps understanding why CLD still has high concentrations in surface water. The approach proposed here can be used in other Caribbean islands that are poorly equipped for explaining pesticide occurrences in surface waters.