Lab Matters Fall 2024 | Page 133

APHL 2024 POSTER ABSTRACTS concern in these specific communities , assisting in shaping public health initiatives to address this growing concern in rural areas .

Presenter : Doris Yoong Wen Di , doris . di . nsw @ doh . hawaii . gov

In-line Sodium Sulfate Drying to Improve Reduced Sample Volume Extraction Efficiency for US EPA Method 1664B

D . Bissonnette , Biotage

Removal of fats , oils and grease from wastewater is imperative to maintaining the integrity of our water lines and water treatment facilities . The US Environmental Protection Agency ( EPA ) method 1664B outlines the procedure for extraction and gravimetric determination of fats , oils and grease ( FOG ) in aqueous samples . EPA Method 1664B allows the reduction of sample volumes as long as the laboratory is able to meet all the necessary quality control and method criteria at both the one-liter volume and reduced sample volume . Reducing sample volumes offers a tremendous advantage in reducing labor and cost , as well as less solvent and waste generated from the extraction procedure . This study will be exploring an optimized workflow that provides laboratories with full automation of both standard and reduced sample volumes while staying compliant with EPA Method requirements .

With the utilization of automated solid-phase extraction ( SPE ), we demonstrate the benefits of reduced sample volume extractions accompanied by inline sodium sulfate drying . The method detection limit ( MDL ) in this study calculated as 0.9504 mg / L for one-liter samples and 0.8676 mg / L for the reduced volume samples . The seven replicate samples used for the MDL calculations show little variation keeping the calculated MDL below 1.4 mg / L , the required MDL per EPA Method 1664B . The MDLs were calculated using 6 degrees of freedom and a student t-value of 3.143 with a 99 % confidence interval . The recoveries for each set of four Initial Precision and Recovery ( IPR ) samples are well within the acceptable range of 83-101 % recovery , per EPA Method 1664B . The IPR resulted in a precision of 0.72 . This is well below the method requirement of 11.0 . The data from the one liter and reduced sample volume extractions demonstrates that the recovery of oil and grease on each extraction system is within the necessary recovery range outlined in US EPA Method 1664B , remaining compliant with the quality and consistency requirements .

Presenter : Deanna Bissonnette , deanna . bissonnette @ biotage . com

Integrated Wastewater and Clinical Genomic Surveillance for Strategic Planning of MPOX Vaccination in Southern Nevada

W . Bendik 1 , V . Vo 2 , M . Moshi 2 , C . Chang 2 , A . Young 1 , E . Buttery 1 , H . Zhang 1 , S . Afzal 1 , S . Skies 1 , K . Franich 3 , D . Gerrity 4 , C . Lockett 5 , E . Oh 2 , H . Kan 1 , Southern Nevada Public Health Laboratory 1 , University of Nevada-Las Vegas 2 , SNHD- Disease Surveillance and Control 3 , Southern Nevada Water Authority 4 , Southern Nevada Health District 5

Wastewater-based epidemiology ( WBE ) studies , coupled with whole genome sequencing ( WGS ), were employed to trace MPXV lineages and identify tourist locations in Las Vegas , Nevada . The Southern Nevada Public Health Laboratories ( SNPHL ) collaborated with the University of Nevada , Las Vegas ( UNLV ) WBE project to analyze wastewater samples from eight wastewater treatment plant ( WWTP ) locations and two manholes from the locations at the southern portion of the Las Vegas Strip and from nightclubs were also analyzed for MPXV DNA . Our investigation included the quantification and sequencing of MPXV DNA in samples from both wastewater treatment plants and upstream manhole sites .

Weekly samples were collected throughout 2022 , with nucleic acids extracted immediately using a Promega Wizard Enviro Total Nucleic Acid Kit ( Cat # A2991 ). WGS libraries were constructed using a custom primer panel for multiplex PCR on the MPXV complete genome ( NC _ 063383.1 ). We utilized a pipeline developed by the UNLV bioinformatics team to process the WGS data . Sequencing reads underwent adapter trimming with cutadapt v4.240 . Sequencing reads were then aligned to the MPXV genome ( NC _ 063383.1 ) using bwa mem v0.7.17-r118841 .

Between June and October 2022 , SNPHL analyzed 23 genomes from the 295 confirmed cases in Southern Nevada , revealing a spectrum of clinical genomes corresponding to B . 1 lineages , including B . 1.1 , B . 1.2 , B . 1.5 and B . 1.7 . The wastewater from tourist locations contained B . 1.1 and B . 1.2 lineages . Surprisingly , six distinct mutations were found in MPXV genomes from wastewater up to three months before they were found in clinical cases from Nevada . Moreover , two mutations were found in wastewater samples from Las Vegas around two weeks and six months before they were found in clinical samples from Texas , the USA and South Korea , respectively . In addition , the detection of MPXV DNA from a manhole location closer to the tourist location populated by bars and nightclubs revealed distinct temporal dynamics in comparison to results from the WWTPs . Examination of wastewater genome data also concluded genomic surveillance for potential antiviral or vaccine responsiveness . Tecovirimat , an antiviral treatment for smallpox , was authorized under exceptional circumstances to treat MPOX infections . Tecovirimat targets the F13L gene product VP37 , necessary for virus particle formation . In line with the clinical genomes observed in 2022 , we have discovered a Glu353Lys missense mutation present in all wastewater MPXV genomes .

In conclusion , data collectively demonstrate the invaluable potential of wastewater and public health genomic surveillance of MPXV in tracking the evolutionary patterns of the MPXV genome . This suggests that the examination of wastewater genomes could potentially support genomic surveillance for antiviral or vaccine responsiveness . By combining viral quantification and sequencing , we can strategically prioritize tourist and community locations , optimizing the distribution of limited public health resources . This integrated approach enhances our preparedness and response capabilities against emerging infectious diseases like MPOX .

Presenter : Horng-Yuan Kan , kan @ snhd . org

Leveraging Existing Wastewater Surveillance Infrastructure for Short-term Poliovirus Surveillance — Chicago , March – July 2023

A . Kittner 1 , C . Korban 1 , R . Poretsky 2 , S . Green 3 , K . Kunstman 3 , D . Foulkes 1 , K . Anderson 1 , T . Jeon 3 , H . Barbian 3 , S . Bobrovska 3 , M . Pacilli 1 , Chicago Department of Public Health 1 , University of Illinois at Chicago 2 , Rush University Medical Center 3

Introduction : At the direction of the Centers for Disease Control and Prevention ( CDC ), the Chicago Department of Public Health ( CDPH ) leveraged the city ’ s existing wastewater surveillance system ( WS ) for poliovirus surveillance in spring / summer 2023 . This was in