Natural grasslands represent the second largest ecosystem in Turkey. However, the impact of varying nitrogen (N) fertilization rates on overall soil health indicators have not been reported in the country. A 2-year study was conducted in the Kahramnmaras Plateau region in Turkey to evaluate the impacts of seven N application rates [i.e., 0 (N0), 50 (N50), 100 (N100), 150 (N150), 200 (N200), 250 (N250), and 350 (N350) kg N ha−1] on physical, chemical, and biological parameters of soil health. Nitrogen addition decreased bulk density by 8–12%, and increased aggregate stability by 3–5% and EC up to 110%. Application of ≥ 100 kg N ha−1 increased soil porosity up to 6.7%. Soil pH and C:N ratios were not affected by N addition. The lowest plant available water occurred with the N0 and N50 treatments, decreasing around 24% and 17% compared to N300. Soil organic carbon, total nitrogen, and C and N stocks increased with increasing N addition. Application of N300 rates increased C stocks between 4 and 34%, and N stocks between 15 and 22% compared to all other treatments. Compared to control, N250 increased microbial biomass carbon by 349% and nitrogen by 250%. Microbial respiration in the N250 and the N300 treatments was 97% and 129% greater than control. Addition of N fertilization for a first time in a grassland ecosystem with a previous history of long-term overgrazing, even at low rates, positively impacted several parameters of soil health, a positive impact that could ensure a greater sustainability of these fragil systems over the long-term.
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