Climate change, inter-annual precipitation variability, recurrent droughts, and flash flooding, coupled with increasing water needs, are shaping the co-evolution of socioeconomic and cultural assemblages, water laws and regulations, and equitable drinking water access and allocation worldwide. Recognizing the need for mitigation strategies for drinking water availability in urban areas, the Isotope Hydrology Section of the International Atomic Energy Agency (IAEA) coordinated a state-of-the-art global assessment to evaluate water sources and distribution of drinking water supply in urban centers, an initiative entitled “Use of Isotope Techniques for the Evaluation of Water Sources for Domestic Supply in Urban Areas (2018-2023)”. Here, we report on a) current research trends for studying urban drinking water systems during the last two decades and b) the development, testing, and integration of new methodologies, aiming for a better assessment, mapping, and management of water resources used for drinking water supply in urban settings. Selected examples of water isotope applications (Canada, USA, Costa Rica, Ecuador, Morocco, Botswana, Romania, Slovenia, India, and Nepal) provide context to the insights and recommendations reported and highlight the versatility of water isotopes to underpin seasonal and temporal variations across various environmental and climate scenarios. The study revealed that urban areas depend on a large spectrum of water recharge across mountain ranges, extensive local groundwater extraction, and water transfer from nearby or distant river basins. The latter is reflected in the spatial isotope snapshot variability. High-resolution monitoring (hourly and sub-hourly) isotope sampling revealed large diurnal variations in the wet tropics (Costa Rica) (up to 1.5‰ in δ 18O) and more uniform diurnal variations in urban centers fed by groundwater sources (0.08 ‰ in δ 18O) ([Ljubljana](https://www.google.com/search?client=firefox-b-1-d&sca_esv=f5a20a2e9138d638&sca_upv=1&sxsrf=ADLYWIKR6-DvBtjaWqFYRhn6VgnegOa8kg:1717189104058&q=Ljubljana&stick=H4sIAAAAAAAAAONgVuLQz9U3SMrNNXnEaMwt8PLHPWEprUlrTl5jVOHiCs7IL3fNK8ksqRQS42KDsnikuLjgmngWsXL6ZJUm5WQl5iUCAAFa64FOAAAA&sa=X&ved=2ahUKEwjMrrz047iGAxWyG9AFHSVwCBgQzIcDKAB6BAgTEAE), Slovenia). Similarly, while d-excess was fairly close to the global mean value (+10 ‰) across all urban centers (10-15‰), reservoir-based drinking water systems show significantly lower values (up to ~ -20 ‰) (Arlington, TX, USA and Gaborone, Botswana), as a result of strong evapoconcentration processes. δ 18O time series and depth-integrated sampling highlighted the influence of the catchment damping ratio in the ultimate intake water composition. By introducing new, traceable spatial and temporal tools that span from the water source to the end-user and are linked to the engineered and socio-economic structure of the water distribution system, governmental, regional, or community-based water operators and practitioners could enhance drinking water treatment strategies (including more accurate surface water blending estimations) and improve urban water management and conservation plans in the light of global warming.

Extracting water from discrete xylem and soil samples and continuous (long-term) monitoring of water vapor across the soil-plant-atmosphere continuum remains challenging and under a large debate. Here, we present a detailed one-year study in the Fort Worth Botanic Garden (north-central Texas) to evaluate the analytical robustness of the centrifugation extraction method and understand water sourcing from three common urban tree species (Elderberry, Sambucus canadensis; Cherry Laurel, Prunus caroliniana; and Boxelder Maple, Acer negundo). Xylem (N=110) isotope ratios (δ 18O and δ 2H) are compared to local precipitation (N=498), throughfall (N=33), and soil water (N=105) at different depths (0-38 cm). Complementary soil water samples were obtained from cup suction lysimeters (N=42) (0-38 cm). Soil and xylem water extraction volumes ranged from 100 µL to 7.5 mL in plant samples and from 100 µL to 10.5 mL in soil samples. Extraction success rates were 68.8% and 75.2% for xylem and soil samples, respectively. The minimum sample total water content for effective extractions was determined as 10.6% (soil) and 17.8% (xylem). Xylem mean narrowband and broadband (proxy for organic contamination) were 0.23±0.40 (-) and 1.00±0.01 (-), respectively. These values agree with mean narrowband and broadband metrics from throughfall and soils, which highlight the non-invasive nature of centrifugated extractions. Annual mean soil δ 18O compositions (-3.6±1.7‰) corresponded with the throughfall input (-3.6±2.4‰). Xylem δ 18O compositions exhibited a strong temporal enrichment trend at the end of the winter, summer, and fall seasons. Mean spring xylem δ 18O (-2.85‰) was less variable and close to soil mean compositions (-2.82‰). For this season, Bayesian mixing analysis showed source water contributions from distinct soil depths: 0 cm to 12.7 cm for Boxelder Maple, 12.7 cm to 25.4 cm for Cherry Laurel, and 12.7 cm to 38.1 cm for Elderberry. Our results offer a standardized and effective protocol for centrifugation extractions and reveal plant water uptake preferences in a highly altered urban green space during an unprecedented warm year.