Legionella pneumophila occurrence in reduced-occupancy buildings in 11 cities during the COVID-19 pandemic

In spring 2020, reduced water demand was an unintended consequence of COVID-19 pandemic-related building closures. Concerns arose that contaminants associated with water stagnation, such as Legionella pneumophila, could become prevalent. To investigate this potential public health risk, samples from 26 reduced-occupancy buildings across 11 cities in the United States, Canada, and Switzerland were analyzed for L. pneumophila using liquid culture (Legiolert, n=258) and DNA-based methods (qPCR/ddPCR, n=138). L. pneumophila culture-positivity was largely associated with just five buildings, each of which had specific design or operational deficiencies commonly associated with L. pneumophila occurrence. Samples from free chlorine buildings had higher culture-positivity (37%) than chloramine buildings (1%), and 78% of culture-positive samples occurred when the residual was [≤]0.1 mg/L Cl2. Although overall sample positivities using culture- and DNA-based methods were equivalent (34% vs. 35%), there was disagreement between the methods in 13% of paired samples. Few buildings reported any water management activities, and L. pneumophila concentrations in flushed samples were occasionally greater than in first-draw samples. This study provides insight into how building plumbing characteristics and management practices contribute to L. pneumophila occurrence during low water use periods and can inform targeted prevention and mitigation efforts. Synopsis StatementLegionella pneumophila occurrence was evaluated in reduced-occupancy buildings during the COVID-19 pandemic across multiple cities. Graphic for Table of Contents (TOC) O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=108 SRC="FIGDIR/small/22277022v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@12e4961org.highwire.dtl.DTLVardef@1d381eforg.highwire.dtl.DTLVardef@f78342org.highwire.dtl.DTLVardef@1ca856d_HPS_FORMAT_FIGEXP M_FIG C_FIG

SARS-CoV-2 RNA is enriched by orders of magnitude in solid relative to liquid wastewater at publicly owned treatment works

Wastewater-based epidemiology has gained attention throughout the world for detection of SARS-CoV-2 RNA in wastewater to supplement clinical testing. Methods have been developed using both the liquid and the solid fraction of wastewater, with some studies reporting higher concentrations in solids. To investigate this relationship further, we collaborated with six other laboratories to conduct a study across five publicly owned treatment works (POTWs) where both primary solids and raw wastewater influent samples were collected and quantified for SARS-CoV-2 RNA. Solids and influent samples were processed by participating laboratories using their respective methods and retrospectively paired based on date of collection. SARS-CoV-2 RNA concentrations by mass (gene copies per gram) were higher in solids than in influent by approximately three orders of magnitude. Concentrations in matched solids and influent were positively and significantly correlated at all five POTWs. RNA concentrations in both solids and influent were correlated to COVID-19 incidence rates in the sewershed and thus representative of disease burden; the solids methods appeared to produce a comparable relationship between SARS-CoV-2 RNA concentration measurements and incidence rates across all POTWs. Solids and influent methods showed comparable sensitivity, N gene detection frequency, and calculated empirical incidence rate lower limits. Analysis of solids has the advantage of using less sample volume to achieve similar sensitivity to influent methods.