A Metagenomic Investigation of Temporal Changes in Wastewater Microbiome across North Carolina A&T State University Campus
Description
Wastewater microbial communities are highly dynamic, exhibiting significant variation over time. We employed Next-Generation Sequencing to identify various SARS-CoV-2 variants of concern, followed by a shotgun metagenomic approach to examine the relative abundance of pathogens in wastewater associated with SARS-CoV-2 across the North Carolina A&T State University campus. Biweekly grab samples were collected across the NCA&T State University campus between January 2021 and December 2023. RT-qPCR results revealed higher concentrations of SARS-CoV-2 RNA in wastewater samples from 2021-2022 compared to 2023. Targeted sequencing detected all major variants of concern, including Alpha, Delta, and Omicron, in 2021-2022 samples. Additionally, a shotgun metagenomic approach was used to investigate changes in wastewater microbial communities from a COVID-19 quarantine dormitory (Haley Hall) on campus, with samples collected each semester over a three-year period (2021-2023). Results showed a higher prevalence of healthy gut bacteria during the Spring semesters of 2021-2023, while pathogenic bacterial taxa were more prominent in Fall 2023, highlighting the impact of pathogens on the sewage microbiome. However, no significant temporal variation in the overall prokaryotic community composition was observed between or within Spring and Fall semesters from 2021 to 2023 using alpha and beta diversity indices (p >0.05). These findings underscore the dynamic nature of the wastewater microbiome