Monitoring Lyme Disease in a Small Urban City Through Wastewater Detection of <i>Borrelia burgdorferi</i>

Lyme disease remains the most reported vector-borne disease in the United States, yet traditional surveillance methods rely heavily on clinical diagnosis and laboratory confirmation, both of which are subject to underreporting and diagnostic limitations. This study evaluated the feasibility of wastewater-based epidemiology (WBE) as a complementary surveillance tool for monitoring the lyme disease causing agent, Borrelia burgdorferi in a small urban community. It was conducted between 2023-2025 with 92 influent wastewater samples collected from two municipal treatment facilities in Bloomington, Indiana. Samples were concentrated using 0.45 µm pore size electronegative membrane filtration and analyzed via digital PCR for B. burgdorferi, with crAssphage quantified as a fecal normalization marker. B. burgdorferi was detected intermittently at low concentrations, with peak values reaching 3,649 gene copies/L. Detection exhibited pronounced seasonal variability, with the highest positivity occurring in fall (62.5%) and no detections observed in spring. No statistically significant differences were observed between treatment plants. Our findings indicate that B. burgdorferi can be detected in municipal wastewater and that detection patterns align with known seasonal trends in Lyme disease transmission. Despite low and intermittent detections, WBE may serve as a useful complementary population- level surveillance tool for B. burgdorferi in small urban systems.