The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institution

ABSTRACT Testing for the causative agent of coronavirus disease 2019 (COVID-19), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been crucial in tracking disease spread and informing public health decisions. Wastewater-based epidemiology has helped to alleviate some of the strain o...

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Main Authors: Chequita Brooks, Sebrina Brooks, Josie Beasley, Jenna Valley, Michael Opata, Ece Karatan, Rachel Bleich
Format: Article
Language:English
Published: American Society for Microbiology 2025-02-01
Series:Microbiology Spectrum
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Online Access:https://journals.asm.org/doi/10.1128/spectrum.02003-24
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author Chequita Brooks
Sebrina Brooks
Josie Beasley
Jenna Valley
Michael Opata
Ece Karatan
Rachel Bleich
author_facet Chequita Brooks
Sebrina Brooks
Josie Beasley
Jenna Valley
Michael Opata
Ece Karatan
Rachel Bleich
author_sort Chequita Brooks
collection DOAJ
description ABSTRACT Testing for the causative agent of coronavirus disease 2019 (COVID-19), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been crucial in tracking disease spread and informing public health decisions. Wastewater-based epidemiology has helped to alleviate some of the strain of testing through broader, population-level surveillance, and has been applied widely on college campuses. However, questions remain about the impact of various sampling methods, target types, environmental factors, and infrastructure variables on SARS-CoV-2 detection. Here, we present a data set of over 800 wastewater samples that sheds light on the influence of a variety of these factors on SARS-CoV-2 quantification using droplet digital PCR (ddPCR) from building-specific sewage infrastructure. We consistently quantified a significantly higher number of copies of virus per liter for the target nucleocapsid 2 (N2) compared to nucleocapsid 1 (N1), regardless of the sampling method (grab vs composite). We further show some dormitory-specific differences in SARS-CoV-2 abundance, including correlations to dormitory population size. Environmental variables like precipitation and temperature show little to no impact on virus load, with the exception of higher temperatures for grab sample data. We observed significantly higher gene copy numbers of the Omicron variant than the Delta variant within ductile iron pipes but no difference in nucleocapsid abundance (N1 or N2) across the three different sewage pipe types in our data set. Our results indicate that contextual variables should be considered when interpreting wastewater-based epidemiological data.IMPORTANCETesting for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), has been crucial in tracking the spread of the virus and informing public health decisions. SARS-CoV-2 viral RNA is shed by symptomatic and asymptomatic infected individuals, allowing its genetic material to be detected and quantified in wastewater. Here, we used wastewater-based epidemiology to measure SARS-CoV-2 viral RNA from several dormitories on the Appalachian State University campus and examined the impact of sampling methods, target types, environmental factors, and infrastructure variables on quantification. Changes in the quantification of SARS-CoV-2 were observed based on target type, as well as trends for the quantification of the Delta and Omicron variants by sampling method. These results highlight the value of applying the data-inquiry practices used in this study to better contextualize wastewater sampling results.
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spelling doaj-art-2fd93b34da9b4c92ba1056aafe35eb902025-02-04T14:03:41ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-02-0113210.1128/spectrum.02003-24The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institutionChequita Brooks0Sebrina Brooks1Josie Beasley2Jenna Valley3Michael Opata4Ece Karatan5Rachel Bleich6Department of Biology, Appalachian State University, Boone, North Carolina, USADepartment of Biology, University of North Carolina at Wilmington, Wilmington, North Carolina, USADepartment of Biology, Appalachian State University, Boone, North Carolina, USADepartment of Biology, Appalachian State University, Boone, North Carolina, USADepartment of Biology, Appalachian State University, Boone, North Carolina, USADepartment of Biology, Appalachian State University, Boone, North Carolina, USADepartment of Biology, Appalachian State University, Boone, North Carolina, USAABSTRACT Testing for the causative agent of coronavirus disease 2019 (COVID-19), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been crucial in tracking disease spread and informing public health decisions. Wastewater-based epidemiology has helped to alleviate some of the strain of testing through broader, population-level surveillance, and has been applied widely on college campuses. However, questions remain about the impact of various sampling methods, target types, environmental factors, and infrastructure variables on SARS-CoV-2 detection. Here, we present a data set of over 800 wastewater samples that sheds light on the influence of a variety of these factors on SARS-CoV-2 quantification using droplet digital PCR (ddPCR) from building-specific sewage infrastructure. We consistently quantified a significantly higher number of copies of virus per liter for the target nucleocapsid 2 (N2) compared to nucleocapsid 1 (N1), regardless of the sampling method (grab vs composite). We further show some dormitory-specific differences in SARS-CoV-2 abundance, including correlations to dormitory population size. Environmental variables like precipitation and temperature show little to no impact on virus load, with the exception of higher temperatures for grab sample data. We observed significantly higher gene copy numbers of the Omicron variant than the Delta variant within ductile iron pipes but no difference in nucleocapsid abundance (N1 or N2) across the three different sewage pipe types in our data set. Our results indicate that contextual variables should be considered when interpreting wastewater-based epidemiological data.IMPORTANCETesting for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), has been crucial in tracking the spread of the virus and informing public health decisions. SARS-CoV-2 viral RNA is shed by symptomatic and asymptomatic infected individuals, allowing its genetic material to be detected and quantified in wastewater. Here, we used wastewater-based epidemiology to measure SARS-CoV-2 viral RNA from several dormitories on the Appalachian State University campus and examined the impact of sampling methods, target types, environmental factors, and infrastructure variables on quantification. Changes in the quantification of SARS-CoV-2 were observed based on target type, as well as trends for the quantification of the Delta and Omicron variants by sampling method. These results highlight the value of applying the data-inquiry practices used in this study to better contextualize wastewater sampling results.https://journals.asm.org/doi/10.1128/spectrum.02003-24wastewater-based epidemiologySARS-CoV-2droplet digital PCR
spellingShingle Chequita Brooks
Sebrina Brooks
Josie Beasley
Jenna Valley
Michael Opata
Ece Karatan
Rachel Bleich
The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institution
Microbiology Spectrum
wastewater-based epidemiology
SARS-CoV-2
droplet digital PCR
title The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institution
title_full The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institution
title_fullStr The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institution
title_full_unstemmed The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institution
title_short The influence of environmental factors on the detection and quantification of SARS-CoV-2 variants in dormitory wastewater at a primarily undergraduate institution
title_sort influence of environmental factors on the detection and quantification of sars cov 2 variants in dormitory wastewater at a primarily undergraduate institution
topic wastewater-based epidemiology
SARS-CoV-2
droplet digital PCR
url https://journals.asm.org/doi/10.1128/spectrum.02003-24
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