This study examined the effects of different temperatures (37°C and 45°C) and pH (uncontrolled, 8 and 10) on the co-fermentation of FeCl3, anionic polyacrylamide (a-PAM) settling chemical enhanced primary sedimentation (CEPS), and waste activated sludge (WAS) to produce volatile fatty acids (VFAs), which analyzed through characterization, microbial community, and untargeted metabolic pathways. Further, to investigate the interacting effects, temperatures of 37°C, 45°C, and pHs of 8 and 10 were chosen, with a sludge ratio of 7.5. The hydrolysis of sludge was aided by the concentration of soluble chemical oxygen demand (SCOD) rising as temperature and pH rose. Nevertheless, elevated SCOD concentrations failed to materialize into benefits for acid production. Therefore, considering practical applications, the optimal conditions for acid production were determined to be 37°C and pH 10, with a concentration of 3411.17 ± 119.92 mg COD/L. A thorough investigation of correlations between microbial communities, differential metabolites, and environmental factors was carried out, highlighting the significance of pH in relation to microbial and acid production performance. That instance, the pH affects acid generation efficiency more so than temperature parameters. Within the microbial community, there is a considerable positive correlation between several distinct metabolites and Steptococcus, Trichococcus, and Tepidimimicrobium. These metabolites are intimately associated with the materials and energy needed for biological cell production and metabolism. This could be as a result of how environmental influences alter the distribution of microbial populations, which influences various metabolites and ultimately the process of producing acid.
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