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The effects of octanoic acid/nonanoic acid and acclimation time on the synthesis of short-chain-length and medium-chain-length PHA blends from activated sludge were investigated. An increased concentration (847−1366 mg/L) of PHAs resulted from 4-month acclimation compared with the concentration derived from 2-month acclimation (450−1126 mg/ L). The content of octanoic acid had a positive linear relationship with the content of even-numbered carbon monomers among the PHAs. The blending products were identified mainly with scl-PHAs during the 2-month acclimation period and were thereafter dominated by mcl-PHAs until 4 months of acclimation. Thermal properties analysis demonstrated that the products derived from 4-month acclimation were a mixture of scl-PHAs and mcl-PHAs rather than a copolymer of scl-PHAs and mcl-PHAs. High-throughput sequencing results indicated that Pseudofulvimonas, Paracoccus, and Blastocatella were the dominant genera that might be responsible for scl-PHAs production during the 2-month acclimation period, whereas Comamonas and Pseudomonas that were responsible for mcl-PHAs production then became the dominant genera after 4-months acclimation.