Field Evaluation of pBI143 Genetic Marker for Microbial Source Tracking in Urban Surface Waters

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Abstract This study evaluated the sewage-associated plasmid pBI143 using two independent surface water data sets: long-term monitoring from Lake Pontchartrain, New Orleans (2016–2017), and short-term urban sampling at Chicago beaches (2025). pBI143 was compared with sewage-associated Carjivirus (formerly crAssphage) and, where available, enterococci, turbidity, and human adenovirus (HAdV). Conditional probability analysis was used to estimate confidence that sewage pollution was present given pBI143 detection. In New Orleans, pBI143 was detected in 88.3% of samples (53/60), compared with 35.0% (21/60) of Carjivirus. All Carjivirus-positive samples were also pBI143-positive. In Chicago, pBI143 and Carjivirus were detected in 66.7% (24/36) and 52.8% (19/36) of samples, respectively, with partial overlap. pBI143 was positively correlated with Carjivirus in New Orleans (Spearman ρ = 0.545, p < 0.0001), whereas the association was weaker in Chicago (ρ = 0.332, p = 0.048). Agreement analysis indicated weak to moderate concordance between pBI143 and Carjivirus in both waters, emphasizing marker complementarity. Conditional probability analysis indicated high confidence of sewage pollution given pBI143 detection, with posterior probabilities of 98.4–99.1% in New Orleans and 94.3–96.6% in Chicago. These results indicate that pBI143 is a sensitive sewage-associated marker that complements Carjivirus for MST in surface waters.

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