Universal performance of benzalkonium chloride for the preservation of
environmental DNA in seawater samples
Abstract
Environmental DNA (eDNA) analysis allows non-invasive and cost-effective
monitoring of species distribution and composition in aquatic
ecosystems. Benzalkonium chloride (BAC) treatment is an inexpensive and
simple method for preserving macrobial eDNA in water samples, which is
suitable for maximizing both the number of sampling replicates and water
volume. However, its preservation performance has been evaluated in a
limited manner by species-specific assays, targeting short fragments of
mitochondrial DNA in freshwater and brackish ecosystems. Here, we
examined the performance of BAC in preserving eDNA in seawater samples,
targeting different fragment lengths of mitochondrial and nuclear eDNA,
and community information inferred by eDNA metabarcoding. First, we
quantified the time-series changes of Japanese jack mackerel
(Trachurus japonicus) eDNA concentrations in experimental tanks
and inshore seawater to compare the yields and decay rates of eDNA
between BAC treatments. As a result, BAC addition increased the eDNA
yields at the start of the experiment and substantially suppressed the
initial phase of rapid degradation but not the subsequent phase of
slower degradation. In addition, we performed eDNA metabarcoding
targeting fish community, showing that BAC addition suppressed the
decrease in species richness, where the number of fish species hardly
varied throughout the day. Findings of the present and previous studies
indicate high versatility of BAC in preserving qualitative (species
richness) and quantitative (copy number) information on aqueous eDNA
under various environmental conditions. BAC should therefore be used to
minimize the false-negative detection of eDNA, regardless of target
genetic regions, fragment sizes, environmental conditions, and detection
strategies.