Lab Matters Fall 2023 | Page 112

APHL 2023 POSTER ABSTRACTS

Surveillance of SARS-CoV-2 in Wastewater of Ohio : Consolidation of SARS-CoV-2 Testing from Academic , Commercial and Government Labs to Ohio Department of Health Laboratory ( ODHL )

J . Saxena , J . Stiverson , J . Scott , P . Chalasani , S . Anders , E . Leasure ; Ohio Department of Health

Wastewater ( WW ) -based epidemiology is an effective tool for early detection of SARS-CoV-2 in communities before it can be clinically detected . In May 2020 , “ Ohio Coronavirus Wastewater Monitoring Network ” was started at the Governor ’ s request to provide communities information on SARS-CoV-2 in WW to proactively prevent outbreaks . Routine WW monitoring of SARS- CoV-2 in Ohio started in July 2020 . Seventy-seven WW treatment facilities participated in the network and 8 academic , commercial and government labs performed the SARS-CoV-2 testing via different methodologies ( liquid supernatant or solid fraction of WW ; RT-PCR , digital PCR , and digital droplet PCR ). For an optimal comparison of WW data , a standard method of SARS-CoV-2 testing was needed . Hence , all WW testing sites were planned to be transferred to Ohio Department of Health Laboratory ( ODHL ). By July 2022 , 72 WW sites ( few sites dropped ) were being tested at ODHL . The GT-Molecular surveillance guide was used for virus filtration , concentration ( Innovaprep Concentrating Pipette ; ICP ), RNA extraction and SARS-CoV-2 quantification ( digital PCR amplification of N2 region of the SARS-CoV-2 nucleocapsid gene ). Bovine Coronavirus ( BCoV ) was used as a matrix recovery control . A pilot study was performed at ODHL from October 2021 to December 2021 for testing of SARS-CoV-2 from 4 WW sites ( APHL 2022 poster # 86 ). In our pilot study , the ICP concentration method showed a significant correlation ( r = 0.86 ; P = 20 to r = 0.65 , P = 30 ). In conclusion , ODHL successfully consolidated the WW testing sites using Ceres concentration method to monitor COVID cases in Ohio .

Presenter : Jyotisna Saxena , jyotisna . saxena @ odh . ohio . gov

Wastewater Surveillance of SARS-CoV-2 in South Carolina

K . R . l . Gilmore1 , J . Bonaparte2 , M . Davis2 , C . Greenwood2 , C . Aroh2 , C . J . Weaver2 , N . Epie2 ; 2Association of Public Health Laboratories , 2South Carolina Department of Health & Environmental Control

Introduction : Wastewater surveillance studies are used to track the real-time presence of SARS-CoV-2 , the virus that causes COVID-19 . This surveillance is important because it serves as a more expedient surveillance system in comparison to tracking of positive COVID-19 results . People infected with SARS-CoV-2 can shed the virus in their feces , even if they don ’ t have symptoms ; thus , the virus can then be detected in wastewater and capture viral presence of those who are both asymptomatic and symptomatic . This leads to quicker warnings of COVID-19 spread in communities and health regulations and statistics can be shared quicker and more precisely . Testing for SARS-CoV-2 in wastewater has several technical challenges . One issue is determining the most effective method of wastewater extraction which is crucial to maintaining accurate surveillance ( CDC , 2022 ). Currently , pasteurizing wastewater samples to reduce the chances of viral transmission while testing , while maintaining useful results are an area of interest . Pasteurizing wastewater samples will allow us to perform the extraction process without using the biosafety level 3 ( BSL3 ) guidelines . The purpose of this research is to determine if pasteurizing wastewater samples prior to extraction yields comparable results to non-pasteurized samples .

Methods : Wastewater samples received from two South Carolina counties , Georgetown and York County , with the highest observed concentrations of COVID-19 , were selected for pasteurization testing . 40mL of each sample were aliquoted for pasteurization as well as non-pasteurization . Pasteurization tests were done twice , 1 hour pasteurized samples compared to non-pasteurized samples and 30-minute pasteurized samples compared to non-pasteurized samples in a 65 ° C water bath . After extracting the pasteurized and non-pasteurized samples , quantification was conducted by digital PCR . The results from this procedure will determine if the pasteurization process will be validated by the South Carolina Department of Health and Environmental Control laboratory .

Results : For both Georgetown and York County , pasteurization methods have not been proven to to yield comparable results to non pasteurized wastewater samples for Sars-CoV-2 surveillance .

Conclusion : Thus far , the pasteurization results from the one-hour pasteurization method have been proven to be ineffective . The use of pasteurization on wastewater samples are unnecessary if there is significant viral loss for testing . For both counties , there is roughly a 4.5 decrease in antibody detection level of Sars-CoV-2 of the pasteurized samples compared to the non-pasteurized samples . The results for the 30-minute pasteurization method are still pending , there is hope that with a reduction of pasteurization time there will be less viral loss for wastewater samples .

Presenter : Kirsti Gilmore , gilmorkr @ dhec . sc . gov

Wastewater Surveillance Reports and Analysis of SARS- CoV-2 Concentrations and Variants in Hawai ’ i

M . Steadmon , B . Sze , L . Sylne , E . Desmond , D . Strange , D . Ornellas , S . Remigio , S . Badil ; Hawai ’ i State Department of Health

Wastewater surveillance has grown worldwide since the beginning of the COVID-19 pandemic to become a valuable layer for monitoring community levels of SARS-CoV-2 . Monitoring wastewater is especially vital to the State of Hawai ’ i to understand disease patterns and reduce mortalities , especially in the Native Hawaiian and Pacific Islander communities that consisted of 22 % of COVID-19 cases and deaths . Beginning in Spring 2022 , the Hawai ’ i State Department of Health ( HDOH ) joined the National Wastewater Surveillance System ( NWSS ) and enrolled 15 wastewater treatment facilities across the state to quantify SARS-CoV-2 in sewage . To monitor potential surges , identify lineages spreading in the community , and assist in public health responses , the HDOH publishes reports on the data provided by NWSS on the SARS-CoV-2 concentrations and variants present in wastewater . Surveillance has demonstrated that wastewater SARS-CoV-2 concentrations in sewage strongly correlates with COVID-19 case counts in Hawai ’ i . To use as an early warning indicator , concentrations are plotted over time and fit to a local regression to easily observe changes in trends that could indicate surges . Results have demonstrated that upward or downward trends have often reflected synonymous changes in COVID-19 case counts . As of January 2023 , over 400 SARS-CoV-2 variants have been detected in Hawai ’ i wastewater since surveillance began with detection of variants in sewage often occurring prior to clinical detection . Notably , XBB . 1.5 was identified in Honolulu County wastewater prior to detection in clinical samples . Overall , creating public reports of wastewater surveillance data for