THP-AD sludge metagenome - 140C_155C_165Ca_165Cb Targeted loci

Anaerobic digestion is a widely applied biological process for stabilizing wastewater sludge and producing renewable energy in the form of biogas. Thermal hydrolysis processes (THP) are increasingly used as a pretreatment step to improve sludge degradability, enhance biogas production, and reduce pathogen loads. However, changes in THP operating temperature may alter the solubilization of organic matter and the characteristics of substrates entering the digesters, which can influence the structure and stability of microbial communities involved in hydrolysis, acidogenesis, and methanogenesis.This project investigates the impact of full scale THP operating temperatures (140 C, 155 C, and 165 C) on the microbial communities present in downstream mesophilic anaerobic digesters at a municipal water resource recovery facility. Digestate samples were collected during stable operating periods at each temperature condition, including two samples at 165 C to capture potential temporal variation. DNA was extracted and bacterial and archaeal community composition was analyzed using high throughput sequencing of the 16S rRNA gene (V4-V5 region). Amplicon sequence variants (ASVs) were inferred using a denoising workflow, and taxonomy was assigned using curated reference databases.The objective of this project is to characterize how THP temperature affects microbial diversity, relative abundance, and community level resilience in anaerobic digesters. Particular attention is given to whether temperature driven changes in solubilized organic matter correspond to shifts in dominant fermentative, syntrophic, and methanogenic taxa. The study focuses on community structure rather than functional gene profiling, with emphasis on identifying stable core populations that support digester robustness under changing upstream conditions.The dataset provides insight into the adaptability of anaerobic digestion microbiomes in full scale systems and contributes to a broader understanding of microbial responses to thermal pretreatment strategies. The sequences generated here support comparative studies of environmental anaerobic microbiomes and ongoing efforts to optimize sludge treatment processes in wastewater resource recovery facilities.