This study explores changes in pastoral practices in the Jerusalem region (Iron Age II - Late Hellenistic) through a multi-isotope approach (strontium, carbon, oxygen, and nitrogen). Based on the analysis of 135 sheep, goat, and cattle teeth and bone samples from Givati Parking Lot we demonstrate the value of this method in reconstructing past animal husbandry, revealing adaptation and resilience of pastoral communities amidst environmental and socio-political changes. Isotopic analysis indicates local sourcing for most animals, with intriguing outliers from distant regions up to 150 km away, suggesting regional exchange networks. Notably, the Persian period (5th century BCE) exhibits a wider isotope range, implying increased flexibility and exploitation of diverse grazing lands, potentially driven by climate shifts and political upheavals. Conversely, Late Hellenistic (2nd century BCE) livestock display restricted movement, while showcasing a rise in desert caprines, indicative of increased import compared to the Persian era. These findings highlight the dynamism and adaptability of past pastoral communities, adjusting their strategies in response to various pressures. This study opens new avenues for understanding human-environment interactions in the Levant and underscores the power of multi-isotope approaches in unraveling intricate socio-economic and ecological dynamics of the past.