Benzalkonium Chloride Significantly Improves Environmental DNA Detection from Schistosomiasis Snail Vectors in Freshwater Samples

Abstract

Urogenital schistosomiasis, caused by Schistosoma haematobium and transmitted by Bulinus snails, affects approximately 190 million individuals globally and remains a major public health concern. Effective surveillance of snail vectors is critical for disease control, but traditional identification methods are time-intensive and require specialized expertise. Environmental DNA (eDNA) detection using qPCR has emerged as a promising alternative for large-scale vector surveillance. To prevent eDNA degradation, benzalkonium chloride (BAC) has been proposed as a preservative, though its efficacy with schistosomiasis snail vectors has not been evaluated. This study tested the impact of BAC (0.01%) on the stability of Bulinus truncatus eDNA under simulated field conditions. Water samples from aquaria with varying snail densities (0.5–30 snails/L) were stored up to 42 days with BAC. eDNA detection via qPCR and multivariable linear mixed regression analysis revealed that BAC enhanced eDNA stability. eDNA was detectable up to 42 days in samples with ≥1 snail/L and up to 35 days at 0.5 snails/L. Additionally, a positive correlation between snail density and eDNA concentration was observed. These findings support the development of robust eDNA sampling protocols for field surveillance, enabling effective monitoring in remote areas and potentially distinguishing between low- and high-risk schistosomiasis transmission zones.