SARS-CoV-2 virus in Raw Wastewater from Student Residence Halls with concomitant 16S rRNA Bacterial Community Structure changes

Ye Li¹K T Ash¹Dominique Joyner¹D E Williams¹Terry C Hazen^2,3,4,5*

• ¹Department of Civil and Environmental Engineering, Tickle College of Engineering, University of Tennessee, Knoxville, Knoxville, Tennessee, United States
• ²Department of Civil & Environmental Engineering, The University of Tennessee, Knoxville, Knoxville, United States
• ³Biosciences Division, Oak Ridge National Laboratory (DOE), Oak Ridge, Tennessee, United States
• ⁴Department of Microbiology, College of Arts and Sciences, The University of Tennessee, Knoxville, Knoxville, Tennessee, United States
• ⁵Department of Earth and Planetary Sciences, College of Arts and Sciences, The University of Tennessee, Knoxville, Knoxville, Tennessee, United States

The detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in sewage is well-established, but the concomitant changes in microbial compositions during the pandemic remain insufficiently explored. This study investigates the impact of the SARS-CoV-2 virus on microbial compositions in raw sewage, utilizing high-throughput 16S rRNA amplicon sequencing to analyze wastewater samples collected from six dormitories over a one-year field trial at the University of Tennessee, Knoxville. The concentration of SARS-CoV-2 RNA was assessed using a reverse transcription-quantitative polymerase chain reaction. Significant variations in bacterial composition were evident across the six dormitories, highlighting the importance of independently considering spatial differences when evaluating the raw wastewater microbiome. Positive samples for SARS-CoV-2 exhibited a prominent representation of exclusive species across all dormitories, coupled with significantly reduced bacterial diversity compared to negative samples. The correlation observed between the relative abundance of enteric pathogens and potential pathogens at sampling sites introduces a significant dimension to our understanding of COVID-19, especially the notable correlation observed in positive SARS-CoV-2 samples. Furthermore, the significant correlation in the relative abundance of potential pathogens between positive and negative SARS-CoV-2 raw sewage samples may be linked to the enduring effects of microbial dysbiosis observed during COVID-19 recovery. These findings provide valuable insights into the microbial dynamics in raw sewage during the COVID-19 pandemic.The detection of SARS-CoV-2 RNA in sewage is established, yet understanding its impact on microbial compositions is limited. This study investigates this impact in raw sewage using 16S rRNA sequencing at the University of Tennessee, Knoxville. Significant bacterial composition variations were found among dormitories, emphasizing the importance of spatial differences.Positive SARS-CoV-2 samples exhibited distinct species representation and decreased bacterial diversity. Notably, the correlation between the abundance of enteric pathogens and potential pathogens at sampling sites enriches our understanding of COVID-19, particularly evident in positive SARS-CoV-2 samples. Furthermore, the link between the abundance of potential pathogens in positive and negative SARS-CoV-2 samples implies enduring microbial dysbiosis effects during COVID-19 recovery. These findings shed light on raw sewage microbial dynamics during the pandemic, offering invaluable contributions to the field.