Wastewater Treatment Plants (WWTPs) generate a substantial amount of sludge as a residue of the wastewater treatment process. The management of this sludge requires the adoption of cost-effective and environmentally friendly techniques. Sludge Treatment Wetlands (STW) have emerged as an effective means to manage and treat sludge, resulting in the production of biosolids suitable for agricultural purposes. This study aimed to ascertain the optimal operation in terms of final rest period for STWs in the Mediterranean region to boost the production of biosolids for agricultural applications. Two WWTPs in Spain were monitored for a period of 9 months, during which several physic-chemical parameters were monitored to determine the optimal final rest period. The findings revealed that, when sludge feeding period was stopped in summer, a final rest period of 6 months was sufficient to reach biosolids suitable for agricultural re-use. These biosolids displayed a content of 53% total solids (TS), a mineralization rate of 40% volatile solids (VS/TS), a stabilization degree (SD) of 40%, and a high nutrient content. Moreover, the presence of heavy metals, pathogens, antimicrobial resistance genes (ARGs), antibiotics, and pesticides was generally low. Overall, this study demonstrates that, in the Mediterranean Region, 6 months of rest period starting in summer are appropriate for sludge treatment wetlands to effectively generate biosolids with suitable characteristics to be safely re-used as a bio-based fertilizer.