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Conventional wastewater treatment plants remove phosphorus, which is captured in sewage sludge. Increasing attention is paid to suitable process pathways that allow recovery and recycling of phosphorus. One of the processes under investigation is acid leaching and recovery of phosphorus, but this requires considerable chemical additives, which could be avoided by stimulating acidification via microbiological processes. This study investigated phosphorus leaching from sewage sludge by biogenic sulfuric acid, using Acidithiobacillus thiooxidans. Sulfur supplementation and solid to liquid ratio were varied to examine how these factors affected phosphorus leaching yield. Chemical leaching by sulfuric acid from sewage sludge and thermally-treated sludge was conducted to compare with bioleaching from sewage sludge. Sewage sludge samples were collected from wastewater treatment plants in Ghent, Belgium, and Delft, The Netherlands. Both bioleaching and chemical leaching were conducted at laboratory scale using shake flask technique, and highest phosphorus leaching yield and time was determined using one-way ANOVA statistical tests. Biogenic sulfuric acid produced by A. thiooxidans extracted phosphorus from both sludge samples. The highest phosphorus leaching yield observed was 48 ± 0% for 17 days from Ghent samples and 57 ± 4% for 27 days from Delft samples with 5.0% (w/v) sulfur supplementation and 1.0% (w/v) solid to liquid ratio. Chemical leaching took shorter than bioleaching, but the leaching yield was lower, i.e. 41 ± 1% for 4 h from Ghent samples, 44 ± 1% for 1 h from Delft samples, 48 ± 1% for 1 h from thermally-treated Ghent samples and 51 ± 2% for 4 h from thermally-treated Delft samples. During phosphorus bioleaching, pH increase was observed during the early stage which hampered the activity of A. thiooxidans and therefore increased phosphorus leaching time. This study suggests that creating conditions for A. thiooxidans to overcome acid neutralizing capacity of sewage sludge is needed to extract phosphorus effectively., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)