New insights on the hydrochemistry, geothermometry, and isotopic characteristics of the hydrothermal groundwater of the SASS basin: case study of the Jérid geothermal field, Southern Tunisia.

Chemical and isotopic indicators were used to recognize the origin of hydrothermal groundwater, to assess the mineralization processes and groundwater quality, to identify the source of solutes and the likely mixing with cold, and elucidate the fluid geothermometry in the Jérid field of Southern Tunisia. The results show that the geothermal groundwater is neutral to slightly alkaline. They are characterized by SO4-Cl-Na-Ca water type. The dissolution of evaporates and pyrite-bearing rocks is the dominant mineralization process. The groundwater quality index indicates that the majority of samples are very hard and belong to poor to unsuitable for drinking classes. Applications and calculations of hydrogeochemical parameters, including SAR, %Na, PI, Kr, and MAR, showed that the majority of samples are unsuitable for agricultural practices. The human health risk was assessed based on hazard quotient and total hazard index through ingestion and dermal contact with iron-rich groundwater. The consumption of CI groundwaters does not present non-carcinogenic risk to adults and children. The δ18 O and δ2H signatures indicate that the geothermal groundwater was recharged by ocean precipitation during cold and wet paleoclimatic periods. The slight enrichment of oxygen-18 and deuterium contents suggests a limited mixing effect between geothermal water and cold groundwater within the same aquifer. This mixing effect is confirmed by the Na-K-Mg and the chloride-enthalpy diagrams. The K-Mg and SiO2 geothermometers provided fairly reliable reservoir temperature values, ranging between 69.6 and 99 °C. Calculated geothermal potential values, varying between 469 and 16987 kWth, which allow several applications such as domestic and agricultural heating.

Isotope hydrology of the intermontane Elk Valley, British Columbia: an assessment of water resources around coal mining operations.

This study aimed to synthesise and interpret stable isotopic data (δ2H and δ18O) from various sources to understand the isotope hydrology around coal mine operations in Elk Valley, B.C., Canada. The data, including precipitation, groundwaters, seeps, and mine rock drains, were used to construct a local meteoric water line (LMWL) for the Elk Valley, evaluate the spatiotemporal isotopic composition of its groundwater, and assess mine seepage and mine rock drain discharge. The study revealed a robust LMWL relation (δ2H = 7.4 ± 0.2 · δ18O - 4.3 ± 4.1). The groundwater and seep data indicated a winter season bias and a north-south latitudinal gradient, suggesting rapid near-surface groundwater flow without significant post-precipitation evaporation. Porewater isotope samples from unsaturated mine rock piles (MRPs) showed site-specific evaporation patterns, potentially due to convective air flows or exothermic sulphide oxidation. This research revealed the influence of groundwater and meltwater on rock drain discharge. Based on evaporative mass balance calculations, MRPs seasonally contributed ca. 5 %(December base flow) and 22 % (snowmelt) to drain discharge. The findings underscore the value of stable isotope data collections in the Elk Valley to help better define and quantify the hydrology-hydrogeology, including a better understanding of evaporative conditions in MRPs.

Comparison of carbon, nitrogen, and oxygen stable isotope ratios and mercury concentrations in muscle tissues of five beaked whale species and sperm whales stranded in Hokkaido, Japan.

We studied δ13C, δ15N and δ18O values, and total mercury (THg) concentrations in muscle samples from deep-sea predators - five beaked whale species and sperm whales - stranded along the coast of Hokkaido, in the north of Japan in 2010 and 2019. The δ13C, δ15N and δ18O values, THg concentrations, and body length (BL) of Stejneger's beaked whales were similar to those of Hubbs' beaked whales, which belong to the same genus. In contrast, δ13C values, THg concentrations, and BL of Sato's beaked whales were markedly different from those of Baird's beaked whales, which belong to the same genus. Stejneger's and Hubbs' beaked whales living around Hokkaido may compete in their ecological niches, whereas Sato's and Baird's beaked whales may segregate their ecological niches. Although Cuvier's beaked whales and sperm whales belong to different genera and their BLs were significantly different, their δ13C and δ15N values were similar, probably because they can dive and stay in deeper waters than other beaked whale species. The δ13C values in combined samples from all whales increased with increasing BL, probably owing to the larger whale species' dietary preference for squid. The δ13C values in combined samples from all whales were positively correlated with THg concentrations, whereas the δ15N values in the combined samples were negatively correlated. The δ18O values in combined samples from most whales tended to be positively correlated with THg concentrations. These correlations may be explained by a higher THg load from deep-sea feeding than from pelagic feeding and by a feeding shift towards lower trophic levels.

The exploratory dataset of isotopic composition of different water sources across Kazakhstan.

This work presents the dataset of stable water isotopes of oxygen and hydrogen measured in water samples from different sources (precipitation, surface water, groundwater, tap water) across Kazakhstan from 2017 to 2018 and from 2020 to 2023. The dataset includes results on isotopic composition of 399 water samples, namely precipitation: event-based (n = 108), cumulative monthly (n = 22); surface water: lakes, reservoirs, brooks, rivers, channels (n = 175), groundwater: shallow and artesian groundwater, spring (n = 85), tapwater (n = 9). For each sample name of the source, location, latitude, longitude and date of sampling, measurement uncertainty (one standard deviation) are available. The samples were assessed by plotting the data in dual δ18O vs. δ2H isotope space with reference to values found in the published literature and fitting a linear regression equation for Astana (event) precipitation. Overall, this is the first dataset covering wide range of sources across Kazakhstan, which could be used by global and regional water resource assessments and studies such as tracing water sources, hydrograph separation and end-member analyses, isotope mass balance, evapotranspiration partitioning, residence time analysis and groundwater recharge.

A protocol for distilling animal body water from biological samples and measuring oxygen and hydrogen stable isotopes via cavity ring-down spectroscopy.

The application of stable isotope analysis (SIA) to the fields of ecology and animal biology has rapidly expanded over the past three decades, particularly with regards to water analysis. SIA now provides the opportunity to monitor migration patterns, examine food webs, and assess habitat changes in current and past study systems. While carbon and nitrogen SIA of biological samples have become common, analyses of oxygen or hydrogen are used more sparingly despite their promising utility for tracing water sources and animal metabolism. Common ecological applications of oxygen or hydrogen SIA require injecting enriched isotope tracers. As such, methods for processing and analyzing biological samples are tailored for enriched tracer techniques, which require lower precision than other techniques given the large signal-to-noise ratio of the data. However, instrumentation advancements are creating new opportunities to expand the applications of high-throughput oxygen and hydrogen SIA. To support these applications, we update methods to distill and measure water derived from biological samples with consistent precision equal to, or better than, ± 0.1 ‰ for δ17O, ± 0.3 ‰ for δ18O, ± 1 ‰ for δ2H, ± 2 ‰ for d-excess, and ± 15 per meg for Δ17O.

The stable isotope hydrology of Sable Island, Nova Scotia, Canada with implications for evaluating the water budget of wild horses.

We investigated the stable isotope hydrology of Sable Island, Nova Scotia, Canada over a five year period from September, 2017 to August, 2022. The δ2H and δ18O values of integrated monthly precipitation were weakly seasonal and ranged from -66 to -15 ‰ and from -9.7 to -1.9 ‰, respectively. Fitting these monthly precipitation data resulted in a local meteoric water line (LMWL) defined by: δ2H = 7.22 ± 0.21 · δ18O + 7.50 ± 1.22 ‰. Amount-weighted annual precipitation had δ2H and δ18O values of -36 ± 11 ‰ and -6.1 ± 1.4 ‰, respectively. Deep groundwater had more negative δ2H and δ18O values than mean annual precipitation, suggesting recharge occurs mainly in the winter, while shallow groundwater had δ2H and δ18O values more consistent with mean annual precipitation or mixing of freshwater with local seawater. Surface waters had more positive values and showed evidence of isolation from the groundwater system. The stable isotopic compositions of plant (leaf) water, on the other hand, indicate plants use groundwater as their source. Fog had δ2H and δ18O values that were significantly more positive than those of local precipitation, yet had similar 17O-excess values. δ2H values of horsehair from 4 individuals lacked seasonality, but had variations typical to those of precipitation on the island. Differences in mean δ2H values of horsehair were statistically significant and suggest variations in water use may exist between spatially disparate horse communities. Our results establish an important initial framework for ongoing isotope studies of feral horses and other wildlife on Sable Island.

Origin and stability of pit lake water in Baiyinhua, Inner Mongolia, based on hydrochemistry and stable isotopes.

Isotope technology is widely used in geochemical mechanisms analysis; however, studies on the origin of pit lake water by isotopes in coal concentration areas in grassland are rare. In this study, 20 groups of water samples were collected, which were subjected to chemical analysis to determine the hydrogeochemical characteristics of pit lake water. The mechanisms of pit lake water formation and recharge-evaporation were ascertained through principal component analysis and the Rayleigh fractionation model. The results indicate that the phreatic water is least affected by evaporation, followed by confined water, surface water and pit lake water. The ionic composition of surface water, phreatic water and most of the confined water is mainly affected by leaching, some confined water can be recharged by surface or phreatic water; while the ionic composition of pit lake water is dominantly affected by evaporation (69.4 %) and is less affected by groundwater recharge (17.1 %) and human activities (11.5 %). The pit lake water is recharged by precipitation, phreatic water and the lateral runoff of confined water; however, the proportion of phreatic and confined water recharge is small. The evaporative loss of the pit lake water is 40-61 % of the initial water body.

Topographic influence on ecohydrology in volcanic watersheds of the western Pacific monsoon area: evidence from water stable isotope composition of meteoric water, thermal water, and plants.

In Taiwanese volcanic watersheds, we investigated stable water isotopes in meteoric water, plants, and thermal water. Meteoric water exhibited a seasonal cycle, with heavier isotopes in winter and lighter ones in summer, especially in the southern region. The northern monsoon signal lagged the south by two weeks. In the Tatun mountains, young water fractions indicated prevalent old water sources. In the northern watershed, streamwater mainly came from the winter monsoon, while the southern one was influenced by alternating monsoons. Both indices indicated that winter plants depended on summer rainfall. Streamwater and plants had distinct sources in winter, supporting ecohydrological separation. Thermal spring water's d-excess helped identify water-rock interactions, with low d value signaling such interactions. The topographic wetness index showed a higher summer monsoon contribution to southern streamwater but a lower one to plants. The mean linear channel direction significantly affected the monsoon contribution fraction, with northeast-oriented channels vulnerable to northeastward winter monsoons. Finally, we developed a model illustrating hydrological processes on short and long timescales. Our findings enhance our understanding of hydrological disturbances' impact on water resources and ecosystems.

Stable isotope hydrology of surface and ground waters in King George Island, Antarctica.

The region around the tip of the Antarctic Peninsula is warming fast, a situation that will lead to widespread changes in local hydrological cycles. King George Island (KGI) hosts a complex network of lakes and rivers, fed by glaciers, snow and rain, and underlain by thick permafrost. We present here the first study of the stable isotope composition of the surface waters in the ice-free southern peninsulas of KGI. Permafrost samples had the highest δ18O and δ2H values (-6.7 and -50 ‰, respectively), and river waters the lowest (-9.1 and -70 ‰, respectively), with groundwater (-8.2 and -62.7 ‰, respectively), lakes (-8.6 and -66.8 ‰, respectively) and (summer) meltwater (-9 and -69.5 ‰, respectively) having intermediary values. Our results suggest that a clear separation of the various water bodies (permafrost, snow, meltwater, lakes) based on the δ18Owater and δ2Hwater is possible. Further, water in lakes on a W-E transect (i.e. with increased distance from the Bellingshausen Sea) have a general tendency towards lower δ18O (and δ2H) values. The results allow for the establishment of a baseline against which ongoing and future changes of the hydrological cycle could be analysed, and past climate changes be reconstructed.

Spatial and meteorological controls of stable water isotope dynamics of precipitation in Kashmir Valley, Western Himalaya, India.

In the Himalayas, the lives and livelihoods of millions of people are sustained by water resources primarily depending on the moisture brought by Western Disturbances and Indian Summer Monsoon. In the present study, a network of 12 precipitation stations was established across the Kashmir Valley to understand the spatial and meteorological factors controlling precipitation isotopes. Temperature and relative humidity are dominant meteorological factors, whereas altitude, proximity to forest canopy, land use/land cover, windward and leeward sides of the mountains are the main physical factors influencing precipitation isotopes. The study suggests that the Mediterranean Sea and nearby water bodies along with continental recycling are the dominant sources of moisture from October to May, while the Arabian Sea, Bay of Bengal and continental recycling are the main sources of moisture from June to September. However, some precipitation events from October to May collect moisture from the Arabian Sea and some precipitation events from June to September collect moisture from the Mediterranean Sea. The occasional passage of Western Disturbances in summer merging with the Indian Summer Monsoon yields heavy to very heavy precipitation. The study provides a better understanding of complex spatial and meteorological phenomena controlling precipitation isotopes across the Western Himalayas.

Preparation of 18O-labelled azaspiracids for accurate quantitation using liquid chromatography-mass spectrometry.

Azaspiracids (AZAs) are a group of polyether marine algal toxins known to accumulate in shellfish, posing a risk to human health and the seafood industry. Analysis of AZAs is typically performed using LC-MS, which can suffer from matrix effects that significantly impact the accuracy of measurement results. While the use of isotopic internal standards is an effective approach to correct for these effects, isotopically labelled standards for AZAs are not currently available. In this study, 18O-labelled AZA1, AZA2, and AZA3 were prepared by reaction with H218O under acidic conditions, and the reaction kinetics and sites of incorporation were studied using LC-HRMS/MS aided by mathematical analysis of their isotope patterns. Analysis of the isotopic incorporation in AZA1 and AZA3 indicated the presence of four exchangeable oxygen atoms. Excessive isomerization occurred during preparation of 18O-labelled AZA2, suggesting a role for the 8-methyl group in the thermodynamic stability of AZAs. Neutralized mixtures of 18O-labelled AZA1 and AZA3 were found to maintain their isotopic and isomeric integrities when stored at -20 °C and were used to develop an isotope-dilution LC-MS method which was applied to reference materials of shellfish matrices containing AZAs, demonstrating high accuracy and excellent reproducibility. Preparation of isotopically labelled compounds using the isotopic exchange method, combined with the kinetic analysis, offers a feasible way to obtain isotopically labelled internal standards for a wide variety of biomolecules to support reliable quantitation.

Carbon, nitrogen, and oxygen stable isotope ratios of striped dolphins and short-finned pilot whales stranded in Hokkaido, northern Japan, compared with those of other cetaceans stranded and hunted in Japan.

Strandings of striped dolphins (SD) and short-finned pilot whales (PW) in Hokkaido, northern Japan, are rare but have recently increased, probably due to global warming. We quantified δ13C, δ15N, and δ18O in muscles of SD (n = 7) and PW (n = 3) stranded in Hokkaido and compared these values with those in muscles (red meat products) of hunted SD and PW in three areas of central and southern Japan. δ18O in stranded SD, except for the calf, decreased with increasing body length (BL), whereas δ13C increased, with no BL-related changes in δ15N. The variability of δ18O (range of maximum and minimum) was larger in the stranded SD (7.5 ‰) than of the hunted SD in three areas (0.9, 1.9, and 1.4 ‰), whereas that of δ15N was smaller in the stranded SD than in the hunted SD. Similarly, the variability of δ18O was larger in the stranded PW in Hokkaido (3.3 ‰) than in the hunted PW in central Japan (1.4 ‰). The larger variability of δ18O and smaller variability of δ15N in stranded SD imply long-term sojourning in coastal waters and feeding on small amounts of limited prey species at low trophic levels before death.

Preventing drift of oxygen isotopes of CO2-in-air stored in glass sample flasks: new insights and recommendations.

It is known that the oxygen isotope composition of CO2-in-air, when stored over longer time periods in glass sample flasks, tends to drift to more negative values while the carbon isotope composition remains stable. The exact mechanisms behind this drift were still unclear. New experimental results reveal that water already inside the flasks during sampling plays a major role in the drift of the oxygen isotopes. A drying method to remove any water sticking to the inner walls by evacuating the flasks for more than 72 h while heating to 60 °C significantly decreases drift of the oxygen isotopes. Moreover, flasks not dried with this method showed higher differences among drift rates of individual flasks. This is explained through the buildup of H2O molecules sticking to the inner walls. Humidity of the air samples in the flasks as well as surface characteristics will lead to differences among flasks. Results also show that permeability of water is higher through Viton O-ring flask seals than through polychlorotrifluoroethylene (PCTFE) shaft seals, and that the stability of flasks sealed with the latter is significantly better over time.

Regional groundwater flow system characterization of volcanic aquifers in upper Awash using multiple approaches, central Ethiopia.

ABSTRACTCharacterization of the groundwater flow systems is important for sustainable water resource management decision-making. We have used vertical profiles of electrical conductivity (EC) and water temperature taken at 2 m intervals during drilling of 109 boreholes, and samples for stable isotope analysis (δ18O, δ2H) taken from 47 boreholes to characterize groundwater recharge, flow and discharge. 222Rn measurements and piezometric evidence were used to complement results from the EC and stable isotopes. The converging evidence shows that groundwater in the study area is characterized by a mix of two different groundwater flow systems: i) the deep groundwater systems are connected to the regional groundwater flow originating from the highlands, outside the surface water basin, ii) the shallow groundwater systems get recharge from local rains. The local recharge zones are located in highly urbanized and industrialized zones posing risk to recharge reduction and pollution. Therefore, attention should be given to protect groundwater resources from contamination and increase groundwater resilience to climate change.

Automated rapid triple-isotope (δ15N, δ18O, δ17O) analyses of nitrate by Ti(III) reduction and N2O laser spectrometry.

The nitrogen and oxygen (δ15N, δ18O, δ17O) stable isotopic compositions of nitrate (NO3-) are crucial tracers of nutrient N sources and dynamics in aquatic and atmospheric systems. Methods to reduce aqueous NO3- to N2O gas (microbial or Cd method) before 15N and 18O isotope analyses require multi-step conversion or toxic chemicals, and 17O in N2O cannot be disentangled by IRMS due to isobaric interferences. This technical note describes the automation of the stable-isotope analyses of nitrate by coupling the new Ti method with a headspace autosampler and an N2O triple-isotope laser analyzer based on off-axis integrated cavity output spectroscopy. The automation yielded accurate and precise results for routine determinations of δ15N, δ18O, and δ17O values for aqueous nitrate in environmental waters. Systematic corrections were required for cavity pressure, N2O concentration and water vapour content to obtain the highest precision for all three isotopic ratios. For the first time, an automated laser-based system facilitates routine low-cost triple isotope analyses in studies where high-temporal resolution isotope analyses of NO3- are required but have been, until now, cost-prohibitive and time-consuming (e.g. atmospheric N pollution).

Near-infrared reflectance spectroscopy accurately predicted isotope and elemental compositions for origin traceability of coffee.

Stable isotope ratios and trace elements are well-established tools that act as signatures of the product's environmental conditions and agricultural processes; but they involve time, money, and environmentally destructive chemicals. In this study, we tested for the first time the potential of near-infrared reflectance spectroscopy (NIR) to estimate/predict isotope and elemental compositions for the origin verification of coffee. Green coffee samples from two continents, 4 countries, and 10 regions were analysed for five isotope ratios (δ13C, δ15N, δ18O, δ2H, and δ34S) and 41 trace elements. NIR (1100-2400 nm) calibrations were developed using pre-processing with extended multiplicative scatter correction (EMSC) and mean centering and partial-least squares regression (PLS-R). Five elements (Mn, Mo, Rb, B, La) and three isotope ratios (δ13C, δ18O, δ2H) were moderately to well predicted by NIR (R2: 0.69 to 0.93). NIR indirectly measured these parameters by association with organic compounds in coffee. These parameters were related to altitude, temperature and rainfall differences across countries and regions and were previously found to be origin discriminators for coffee.

Hydrochemistry and stable isotopes revealed focused and diffuse recharge processes in the Sonora River basin, Mexico.

A hydro-geochemical characterization was conducted in the northern part of the Sonora River basin, covering an area of 9400 km2. Equipotential lines indicated that groundwater circulation coincided with the surface water flow direction. Based on the groundwater temperature measured (on average ∼21 °C), only one spring exhibited thermalism (51 °C). Electrical conductivity (160-1750 µS/cm), chloride and nitrate concentrations (>10 and >45 mg/L) imply highly ionized water and anthropogenic pollution. In the river network, δ18O values revealed a clear modern meteoric origin. Focused recharge occurred mainly from the riverbeds during the rainy season. During the dry season, diffuse recharge was characterized by complex return flows from irrigation, urban, agricultural, mining, and livestock. Drilled wells (>50 m) exhibited a strong meteoric origin from higher elevations during the rainy season with minimal hydrochemical anomalies. Our results contribute to the knowledge of mountain-front and mountain-block recharge processes in a semi-arid and human-altered landscape in northern Mexico, historically characterized by limited hydrogeological data.

Determination of nitrate sources in a karst plateau reservoir based on nitrogen and oxygen isotopes.

Investigating the sources, migration and proportional contribution of nitrate is essential to effectively protect water quality. δ15N-NO3-, δ18O-NO3- and Stable Isotope Analysis in R (SIAR) were used to qualitatively and quantitatively analyse nitrate sources in the Pingzhai Reservoir water body. The values of δ15N-NO3- and δ18O-NO3- in water vary with season. Soil organic nitrogen and chemical fertilisers are the main sources of nitrate in autumn, while domestic sewage and livestock manure are the primary sources of nitrate in winter and spring. The SIAR results showed that chemical fertilisers, livestock manure, sewage, and soil organic nitrogen had the highest proportional contribution. In autumn, the proportional contribution of chemical fertilisers to river and reservoir were 47 and 51 %. During winter, the proportional contributions of livestock manure and sewage to river and reservoir were 53 and 68 %, respectively, and in spring 49 and 68 %, respectively. Considering the fragility of karst ecosystems, strict measures should be formulated for the use of chemical fertilisers and standards for sewage discharge should be raised. Control nitrogen input from agricultural activities and prevent water quality deterioration.

Nitrate isotopes (δ15N, δ18O) in precipitation: best practices from an international coordinated research project.

Stable isotope ratios of nitrogen and oxygen (15N/14N and 18O/16O) of nitrate (NO3-) are excellent tracers for developing systematic understanding of sources, conversions, and deposition of reactive atmospheric nitrogen (Nr) in the environment. Despite recent analytical advances, standardized sampling of NO3-) isotopes in precipitation is still lacking. To advance atmospheric studies on Nr species, we propose best-practice guidelines for accurate and precise sampling and analysis of NO3- isotopes in precipitation based on the experience obtained from an international research project coordinated by the International Atomic Energy Agency (IAEA). The precipitation sampling and preservation strategies yielded a good agreement between the NO3- concentrations measured at the laboratories of 16 countries and at the IAEA. Compared to conventional methods (e.g., bacterial denitrification), we confirmed the accurate performance of the lower cost Ti(III) reduction method for isotope analyses (15N and 18O) of NO3- in precipitation samples. These isotopic data depict different origins and oxidation pathways of inorganic nitrogen. This work emphasized the capability of NO3- isotopes to assess the origin and atmospheric oxidation of Nr and outlined a pathway to improve laboratory capability and expertise at a global scale. The incorporation of other isotopes like 17O in Nr is recommended in future studies.

Isotopically labeled ozone: A new approach to elucidate the formation of ozonation products.

As ozonation becomes a widespread treatment for removal of chemicals of emerging concern from wastewater treatment plant effluents, there are increasing concerns regarding the formation of ozonation products (OPs), and their possible impacts on the aquatic environment and eventually human health. In this study, a novel method was developed that utilizes heavy oxygen (18O2) for the production of heavy ozone ([18O1]O2, [18O2]O1, [18O3]) to actively label OPs from oxygen transfer reactions. To establish and validate this new approach, venlafaxine with a well-described oxygen transfer reaction (tertiary amine -> N-oxide) was chosen as a model compound. Observed 18O/16O ratios in the major OP venlafaxine N-oxide (NOV) correlated with expected 18O purities based on tracer experiments. These results confirmed the successful labeling with heavy oxygen and furthermore demonstrate the potential to monitor NOV as an indicator of 18O/16O ratios during ozonation. As a next step, 18O/16O ratios were used to elucidate the formation mechanism of previously described OPs from sulfamethoxazole (SMX). Seven OPs were detected including the frequently described nitro-SMX, which was formed with a maximum yield of 3.2% (of initial SMX). With the successful labeling of six of the seven OPs from sulfamethoxazole, it was possible to confirm their previously proposed formation pathways, and to distinguish oxygen transfer from electron transfer reactions. 18O/16O ratios in OPs indicate that hydroxylation of the aromatic ring and formation of nitro-groups mostly follows oxygen transfer reactions, while electron transfer reactions initiate the formation of hydroxylamine and the abstraction of NH2 leading to catechol.