To transfer or not to transfer: the dilemma of mosaic embryos - a narrative review.

A frequent finding after preimplantation genetic diagnostic testing for aneuploidies using next-generation sequencing is an embryo that is putatively mosaic. The prevalence of this outcome remains unclear and varies with technical and external factors. Mosaic embryos can be classified by the percentage of cells affected, type of chromosome involvement (whole or segmental), number of affected chromosomes or affected cell type (inner mass cell, trophectoderm or both). The origin of mosaicism seems to be intrinsic as a post-zygotic mitotic error, but some external factors can play a role. As experience has increased with the transfer of mosaic embryos, clinical practice has gradually become more flexible in recent years. Nevertheless, clinical results show lower implantation, pregnancy and clinical pregnancy rates and higher miscarriage rates with mosaic embryo transfer when compared with the transfer of euploid embryos. Prenatal diagnosis is highly recommended after the transfer of mosaic embryos. This narrative review is intended to serve as reference material for practitioners in reproductive medicine who must manage a mosaic embryo result after preimplantation genetic testing for aneuploidies.

Antibiotics, antibiotic resistance and associated risk in natural springs from an agroecosystem environment.

This study investigates the occurrence, transport, and risks associated to antibiotic residues, antibiotic resistance genes (ARGs) and antibiotic resistant Escherichia coli (AR-E. coli) in eleven natural springs in an agroecosystem environment with intense livestock production, where groundwater nitrate concentration usually sets above 50 mg L-1. Out of 23 multiple-class antibiotics monitored, tetracycline and sulfonamide residues were the most ubiquitous, and they were detected at concentrations ranging from ng L-1 to µg L-1. Five ARGs were monitored, conferring resistance to the antibiotic classes of major use in livestock production. Thus, genes conferring resistance to sulfonamides (sul1 and sul2) and tetracyclines (tetW) as well as a gene proxy for anthropogenic pollution (intI1) were present in most springs. sul1 was the most abundant, with absolute concentrations ranging from 4 × 102 to 5.6 × 106 gene copies L-1 water. AR-E. coli showing resistance to sulfonamides and tetracyclines was also detected, with a prevalence up to approximately 40 % in some sites but with poor correlations with the concentration of antibiotic residues and ARGs. The occurrence of antibiotics, ARGs and AR-E. coli was characterized by large seasonal variations which were mostly associated to both hydrological factors and reactive transport processes. Finally, a risk assessment approach pointed out towards low risk for both the groundwater environment and human health, when spring water is used for direct human consumption, associated with the occurrence of antibiotics, ARGs and AR-E. coli. However, long-term effects cannot be neglected, and proper actions must be taken to preserve groundwater quality.

Groundwater antibiotic pollution and its relationship with dissolved organic matter: Identification and environmental implications.

The occurrence of veterinary antibiotics and hydro-chemical parameters in eleven natural springs in a livestock production area is evaluated, jointly with the characterization of their DOM fingerprint by Orbitrap HRMS. Tetracycline and sulfonamide antibiotics were ubiquitous in all sites, and they were detected at low ng L-1 concentrations, except for doxycycline, that was present at µg L-1 in one location. DOM analysis revealed that most molecular formulas were CHO compounds (49 %-68 %), with a remarkable percentage containing nitrogen and sulphur (16 %-23 % and 11 %-24 %, respectively). Major DOM components were phenolic and highly unsaturated compounds (~90 %), typical for soil-derived organic matter, while approximately 11 % were unsaturated aliphatic, suggesting that springs may be susceptible to anthropogenic contamination sources. Comparing the DOM fingerprint among sites, the spring showing the most different profile was the one with surface water interaction and characterized by having lower CHO and higher CHOS formulas and aliphatic compounds. Correlations between antibiotics and DOM showed that tetracyclines positively correlate with unsaturated oxygen-rich substances, while sulfonamides relate with aliphatic and unsaturated oxygen-poor compounds. This indicates that the fate of different antibiotics will be controlled by the type of DOM present in groundwater.

Governance and groundwater modelling: Hints to boost the implementation of the EU Nitrate Directive. The Lombardy Plain case, N Italy.

The EU Nitrate Directive has been ruling for almost 30 years, nevertheless nitrate concentration in the Lombardy Plain did not decrease. Together with failures of management implementation, a possible cause for such field observations is that management actions were taken without adequately considering the actual hydrogeological dynamics. To consider this aspect, the paper presents a groundwater flow and transport numerical model of a specific area of the Lombardy Plain. The aim of this model is to demonstrate how modelling, as a management tool, can be useful in the governance process. The groundwater model, using well-known MODFLOW-MT3D codes, is based on existing hydrogeological information, while a nitrogen mass balance has been performed at municipal scale to determine the agricultural N surplus to the subsurface. The model adequately reproduces head levels and nitrate concentrations in observation wells for a 10-year simulation period, showing that 4.5% of the N annual input remains stored in the system. The model indicates the efficiency of rivers and springs to export N out from the system at an estimated rate of 77.5% of the annual N inputs. Back to governance, the model shows that management data at municipal level (e.g. irrigation rates, groundwater withdrawal, N net recharge) provide a satisfactory scale for successfully reproducing nitrate evolution. Hence those variables that can be object of debate during a governance process can be treated as input data to the numerical model. Therefore, backcasting exercises can be conducted to check whether the model outcome fits with the expected results of specific management actions. The model highlights how the N mass balance evolves, providing clues on which factors can be managed to reduce nitrate concentrations and meet the Directive's requirements. Numerical groundwater models, as an option to address water management issues, ultimately contribute to solve the information and capacity governance gaps.

Forecasting nitrate evolution in an alluvial aquifer under distinct environmental and climate change scenarios (Lower Rhine Embayment, Germany).

When investigating future nitrate (NO3-) concentrations in groundwater, climate change has a major role as it determines the future water budget and, in turn, the conditions in the aquifer which will finally have a decisive effect on NO3- concentrations. In this study, the different effects on water balance and NO3- concentration under three projected climate scenarios - RCP 2.6, RCP 4.5, and RCP 8.5 - are analysed in a water protection area in the Lower Rhine Embayment in Germany. Recharge values were calculated from downscaled precipitation and temperature data for the 21st century in a water budget that considers land use in the evapotranspiration term. Nitrate concentration evolution is estimated using recharge and expected fertilization rates with a lumped-parameter model. In order to be able to map the NO3- concentration, the investigation area is divided into 1000 × 1000 m cells. Each cell is assigned a specific NO3- input and a NO3- degradation capacity. Results show significant variations in NO3- development projected with the different climate scenarios due to different temperatures and consequently actual ET, and precipitation. Nevertheless, nitrate concentrations clearly increase in all projections. The total NO3- mass increases most strongly with RCP 8.5 until 2099 (by 89% compared to 2020) and least with RCP 4.5 (by 50%). Further projections show a 20% reduction in agricultural NO3- input can reduce NO3- concentrations, but insufficiently to comply with drinking water guidelines in all regions and aquifers. The model indicates that NO3- input loads should be defined according to future recharge variations governed by climate change. Consequently, a time-varying fertilization rate specific for each region, with their own turnover time and degradation rate, must be estimated to meet pollution environmental goals.

EU Nitrates Directive, from theory to practice: Environmental effectiveness and influence of regional governance on its performance.

Despite the European Nitrate Directive (ND) being issued almost 30 years ago, groundwater nitrate contamination is still a serious threat to ecosystems and human health. In one of the areas most affected by nitrates, the Lombardy Plain (Italy), the effectiveness of the ND and the capacity of governance to support its application correctly was assessed using a socio-hydrogeological approach. Nitrate trends over 11 years show that most regions present steady or increasing concentrations, highlighting how contamination can affect previously impaired situations and supposedly resistant and resilient aquifers. Stakeholder network analysis reveals that the governance framework does not support knowledge dissemination and changes in farmers' attitudes, hindering water quality improvements. Nitrogen input needs to be reduced and manure relocation monitored. The local governance scale has a key role in enhancing ND dissemination. Reports to the EU Commission should integrate multi- or interdisciplinary evaluation of trends, including governance dynamics, alongside hydrochemical information.

Sorption properties and behaviour at laboratory scale of selected pharmaceuticals using batch experiments.

Despite the increasing public concern about the frequent occurrence of pharmaceuticals in the water bodies, their transport and fate are not yet well known; in particular in groundwater. In this laboratory study, batch experiments were conducted to investigate the sorption behaviour of selected pharmaceuticals. The choice of compounds was motivated by their chemical properties as well as by their occurrence at the relevant field sites. It included: antipyrine, atenolol, caffeine, carbamazepine, ciprofloxacin, diclofenac, ketoprofen, ofloxacin, and sulfamethoxazole. Sorption behaviour has already been investigated for some of these compounds (e.g. carbamazepine), but for the others (e.g. antipyrine, ketoprofen), extensive studies are missing so far. For the experiments, artificial and actual aquifer materials from complementary field sites were selected: technical coarse quartz sand and sediments from alluvial Vistrenque Aquifer, France (sandy loam), and fluvio-deltaic Baix Fluvià Aquifer, Spain (sandy clay loam, clay, and medium sand). In these field sites occurrence of pharmaceuticals in groundwater was previously stated, and the presented laboratory experiments were complementary to the field investigations. Five concentration steps for determining the sorption isotherms were investigated. Correlation analysis showed dependencies of K-coefficients of individual compounds and sediment properties; however, no clear, universal patterns for all compounds were found. Batch experiments suggest that sorption behaviour was governed by compound-specific properties rather than by sediment properties. These results contribute to the understanding sorption behaviour of pharmaceuticals in heterogeneous sediments, although some inconsistencies were revealed between laboratory scale results and field scale observations.

Veterinary pharmaceuticals and antibiotics in manure and slurry and their fate in amended agricultural soils: Findings from an experimental field site (Baix Empordà, NE Catalonia).

The fate and transport of 34 veterinary pharmaceuticals (PhACs) is investigated in swine slurry and dairy cattle manure-amended agricultural soils, from an experimental field site, by using both analytical and modelled data. Potential differences on PhACs fate, attributed to the application of distinct swine slurry fractions (total, solid, and liquid), are herein assessed for the first time. Surface and deep soil layers, up to a depth of 120 cm, were analyzed at different periods after an annual fertilization event. Using input data representing typical agricultural soil conditions and the PhACs concentration measured in organic fertilizers the transport of these pollutants was modelled for a period of 10 years, including the monitored annual fertilization event. Fluoroquinolone, tetracycline and pleuromutilin antibiotics, together with anti-helmintics and analgesic and anti-inflammatories, were detected in manure-amended soils, at average concentrations ranging from 0.078 to 150 µg/kg dw in surface layers, with the highest levels found in the fields fertilized with the swine slurry solid fraction. Even though severe disagreements were observed between experimental and simulated PhACs concentrations along the soil column, both approaches pointed out that target compounds strongly adsorb onto surface layers, showing limited mobility along the soil profile. Thus, repeated manure and slurry fertilizations will contribute in building up persistent PhACs residues in the uppermost layers of the soil, while leaching will be a minor process governing their fate towards the subsurface. The ecotoxicological risks posed by the occurrence of PhACs in soils were estimated to be low for terrestrial organisms. Nevertheless the antibiotic enrofloxacin showed some potential to induce negative effects to crops.

Groundwater nitrate pollution and climate change: learnings from a water balance-based analysis of several aquifers in a western Mediterranean region (Catalonia).

Climate change will affect the dynamics of the hydrogeological systems and their water resources quality; in particular nitrate, which is herein taken as a paradigmatic pollutant to illustrate the effects of climate change on groundwater quality. Based on climatic predictions of temperature and precipitation for the horizon of 2021 and 2050, as well as on land use distribution, water balances are recalculated for the hydrological basins of distinct aquifer systems in a western Mediterranean region as Catalonia (NE Spain) in order to determine the reduction of available water resources. Besides the fact that climate change will represent a decrease of water availability, we qualitatively discuss the modifications that will result from the future climatic scenarios and their impact on nitrate pollution according to the geological setting of the selected aquifers. Climate effects in groundwater quality are described according to hydrological, environmental, socio-economic, and political concerns. Water reduction stands as a major issue that will control stream-aquifer interactions and subsurface recharge, leading to a general modification of nitrate in groundwater as dilution varies. A nitrate mass balance model provides a gross estimation of potential nitrate evolution in these aquifers, and it points out that the control of the fertilizer load will be crucial to achieve adequate nitrate content in groundwater. Reclaimed wastewater stands as local reliable resource, yet its amount will only satisfy a fraction of the loss of available resources due to climate change. Finally, an integrated management perspective is necessary to avoid unplanned actions from private initiatives that will jeopardize the achievement of sustainable water resources exploitation under distinct hydrological scenarios.

Isotope and microbiome data provide complementary information to identify natural nitrate attenuation processes in groundwater.

Natural attenuation processes alleviate the impact of fertilization practices on groundwater resources. Therefore, identifying the occurrence of denitrification has become a requirement for water quality management. Several approaches are useful for this purpose, such as isotopic and microbiological methods, each of them providing distinct but complementary information about denitrification reactions, attenuation rates and their occurrence in the aquifer. In this paper, we investigate the contribution of both approaches to describe denitrification in a consolidated rock aquifer (limestone and marls), with a porosity related to fracture networks located in the northeastern sector of the Osona basin (NE Spain). Isotopic methods indicated the origin of nitrate (fertilization using manure) and that denitrification occurred, reaching a reduction of near 25% of the nitrate mass in groundwater. The studied area could be divided in two zones with distinct agricultural pressures and, consequently, nitrate concentrations in groundwater. Denitrification occurred in both zones and at different levels, indicating that attenuation processes took place all along the whole hydrogeological unit, and that the observed levels could be attributed to a larger flow path or, in a minor extent, to mixing processes that mask the actual denitrification rates. Microbiological data showed a correlation between denitrifier genes and the isotopic composition. However, the groundwater microbiome and the distribution of denitrifying bacteria did not reveal a major influence on the denitrification level observed by isotopic methods. This focuses the interest of microbiological analysis to identify functional genes within the bacteria present in the aquifer. Results indicated that isotopic methods provide information of the overall denitrification ability of the hydrogeological unit, and that genomic data represent the processes actually acting nearby the well. A combination of both approaches is advised to support induced in situ attenuation actions in polluted sites.

Characterizing sources and natural attenuation of nitrate contamination in the Baix Ter aquifer system (NE Spain) using a multi-isotope approach.

Nitrate pollution is a widespread issue affecting global water resources with significant economic and health effects. Knowledge of both the corresponding pollution sources and of processes naturally attenuating them is thus of crucial importance in assessing water management policies and the impact of anthropogenic activities. In this study, an approach combining hydrodynamic, hydrochemical and multi-isotope systematics (8 isotopes) is used to characterize the sources of nitrate pollution and potential natural attenuation processes in a polluted basin of NE Spain. δ2H and δ18O isotopes were used to further characterize the sources of recharge of the aquifers. Results show that NO3- is not homogeneously distributed and presents a large range of concentrations, from no NO3- to up to 480mgL-1. δ15N and δ18O of dissolved NO3- identified manure as the main source of nitrate, although sewage and mineral fertilizers can also be isotopically detected using boron isotopes (δ11B) and δ34S and δ18O of dissolved sulphate, respectively. The multi-isotope approach proved that natural denitrification is occurring, especially in near-river environments or in areas hydrologically related to fault zones. δ34S and δ18O indicated that denitrification is not driven by pyrite oxidation but rather by the oxidation of organic matter. This could not be confirmed by the study of δ13CHCO3 that was buffered by the entanglement of other processes and sources.

Nitrate pollution of groundwater; all right , but nothing else?

Contamination from agricultural sources and, in particular, nitrate pollution, is one of the main concerns in groundwater management. However, this type of pollution entails the entrance of other substances into the aquifer, as well as it may promote other processes. In this study, we deal with hydrochemical and isotopic analysis of groundwater samples from four distinct zones in Catalonia (NE Spain), which include 5 different aquifer types, to investigate the influence of fertilization on the overall hydrochemical composition of groundwater. Results indicate that intense fertilizer application, causing high nitrate pollution in aquifers, also homogenize the contents of the major dissolved ions (i.e.; Cl(-), SO4(2-), Ca(2+), Na(+), K(+), and Mg(2+)). Thus, when groundwater in igneous and sedimentary aquifers is compared, significant differences are observed under natural conditions for Cl(-), Na(+) and Ca(2+) (with p-values ranging from <0.001 to 0.038), and when high nitrate concentrations occur, these differences are reduced (most p-values ranged between 0.054 and 0.978). Moreover, positive linear relationships between nitrate and some ions are found indicating the magnitude of the fertilization impact on groundwater hydrochemistry (with R(2) values of 0.490, 0.609 and 0.470, for SO4(2-), Ca(2+) and Cl(-), respectively). Nevertheless, the increasing concentration of specific ions is not only attributed to agricultural pollution, but to their enhancing effect upon the biogeochemical processes that control water-rock interactions. Such results raise awareness that these processes should be evaluated in advance in order to assess an adequate groundwater resources management.

Temporal analysis of spring water data to assess nitrate inputs to groundwater in an agricultural area (Osona, NE Spain).

Non-point agricultural pollution is a major concern in groundwater management. To investigate nitrate input to the subsurface through groundwater recharge, thirteen natural springs were sampled. Discharge, electrical conductivity (EC), nitrate concentration, pH value and water temperature were monitored every two weeks from January 2010 till February 2011 at selected springs in the Osona region (NE Spain). Two extensive hydrochemical analyses were also conducted at the beginning and at the end of the survey. Springs are classified in four groups describing their hydrological response, based on discharge, EC and nitrate data. Geostatistical analysis provides an additional insight into the discharge and nitrate temporal pattern. Even though discharge and EC can be related to specific hydrogeological behaviours, nitrate content shows uniform values in most of the springs with only a minor influence from external factors such as rainfall events, fertilisation regimes and geological traits. Such evenness of outflow might be attributed to a homogenisation of the subsurface processes that determine nitrate infiltration after decades of intensive fertilisation using pig manure. Accumulated nitrate mass load and variograms confirm this result. Assuming that spring data are representative of nitrate leaching towards the underlying aquifer, nitrate content of infiltrating recharge in shallow aquifers should therefore show a steady value over time with only small fluctuations due to natural processes. Nevertheless, temporal fluctuations in nitrate content in aquifers could be also attributed to flow regime alterations due to human groundwater withdrawal.

Development of a stream-aquifer numerical flow model to assess river water management under water scarcity in a Mediterranean basin.

Stream flow, as a part of a basin hydrological cycle, will be sensible to water scarcity as a result of climate change. Stream vulnerability should then be evaluated as a key component of the basin water budget. Numerical flow modeling has been applied to an alluvial formation in a small mountain basin to evaluate the stream-aquifer relationship under these future scenarios. The Arbúcies River basin (116 km(2)) is located in the Catalan Inner Basins (NE Spain) and its lower reach, which is related to an alluvial aquifer, usually becomes dry during the summer period. This study seeks to determine the origin of such discharge losses whether from natural stream leakage and/or induced capture due to groundwater withdrawal. Our goal is also investigating how discharge variations from the basin headwaters, representing potential effects of climate change, may affect stream flow, aquifer recharge, and finally environmental preservation and human supply. A numerical flow model of the alluvial aquifer, based on MODFLOW and especially in the STREAM routine, reproduced the flow system after the usual calibration. Results indicate that, in the average, stream flow provides more than 50% of the water inputs to the alluvial aquifer, being responsible for the amount of stored water resources and for satisfying groundwater exploitation for human needs. Detailed simulations using daily time-steps permit setting threshold values for the stream flow entering at the beginning of the studied area so surface discharge is maintained along the whole watercourse and ecological flow requirements are satisfied as well. The effects of predicted rainfall and temperature variations on the Arbúcies River alluvial aquifer water balance are also discussed from the outcomes of the simulations. Finally, model results indicate the relevance of headwater discharge management under future climate scenarios to preserve downstream hydrological processes. They also point out that small mountain basins could be self-sufficient units so long as the response of the main hydrological components to external forces that produce water scarcity, as climate change or human pressures, is appropriately considered in water resource planning.

Analysis of vulnerability factors that control nitrate occurrence in natural springs (Osona Region, NE Spain).

Nitrate pollution is one of the main concerns of groundwater management in most of the world's agricultural areas. In the Osona region of NE Spain, high concentrations of nitrates have been reported in wells. This study uses the occurrence of this pollutant in natural springs as an indicator of the sub-surface dynamics of the water cycle and shows how groundwater quality is affected by crop fertilization, as an approach to determine the aquifer vulnerability. Nitrate concentration and other hydrochemical parameters based on a biannual database are reported for approximately 80 springs for the period 2004-2009. The background concentration of nitrate is first determined to distinguish polluted areas from natural nitrate occurrence. A statistical treatment using logistic regression and ANOVA is then performed to identify the significance of the effect of vulnerability factors such as the geological setting of the springs, land use in recharge areas, sampling periods, and chemical parameters like pH and EC, on groundwater nitrate pollution. The results of the analysis identify a threshold value of 7-8 mg NO(3)(-)/L for nitrate pollution in this area. Logistic regression and ANOVA results show that an increase in EC or a decrease in pH values is linked to the possibility of higher nitrate concentrations in springs. These analyses also show that nitrate pollution is more dependent on land use than the geological setting of springs or sampling periods. Indeed, the specific geological and soil features of the uppermost layers in their recharge areas do not contribute to the buffering of nitrate impacts on aquifers as measured in natural springs. Land use, and particularly fertilization practices, are major factors in groundwater vulnerability.

Analyzing hydrological sustainability through water balance.

The objective of the Water Framework Directive (2000/60/EC) is to assist in the development of management plans that will lead to the sustainable use of water resources in all EU member states. However, defining the degree of sustainability aimed at is not a straightforward task. It requires detailed knowledge of the hydrogeological characteristics of the basin in question, its environmental needs, the amount of human water demand, and the opportunity to construct a proper water balance that describes the behavior of the hydrological system and estimates available water resources. An analysis of the water balance in the Selva basin (Girona, NE Spain) points to the importance of regional groundwater fluxes in satisfying current exploitation rates, and shows that regional scale approaches are often necessary to evaluate water availability. In addition, we discuss the pressures on water resources, and analyze potential actions, based on the water balance results, directed towards achieving sustainable water management in the basin.