What is driving the NF-κB response in environmental water extracts?

In vitro bioassays are increasingly applied for water quality monitoring, with assays indicative of adaptive stress responses commonly included in test batteries. The NF-κB assay is responsive to surface water and wastewater extracts, but the causative compounds are unknown and micropollutants typically found in water do not activate the NF-κB assay. The current study aimed to investigate if co-extracted organic matter and/or endotoxins could cause the NF-κB response in surface water extracts. The effect of model bacterial lipopolysaccharides (LPS) and dissolved organic carbon (DOC) was evaluated in the NF-κB assay both before and after solid-phase extraction (SPE), with 7% effect recovery for LPS and between 7 and 52% effect recovery for DOC observed. The NF-κB response, endotoxin activity, micropollutant concentration and total organic carbon concentration was measured in four surface water extracts. All water extracts showed a response in the NF-κB assay, but the detected micropollutants could not explain the effect. Comparison of predicted bioanalytical equivalent concentrations based on micropollutant, DOC and endotoxin concentrations in surface water with experimental bioanalytical equivalent concentrations suggest that co-extracted endotoxins are the most important drivers of the observed effect, with DOC only having a minor contribution. While in vitro bioassays typically detect mixtures of organic micropollutants, the current study shows that the NF-κB assay can integrate the effects of co-extracted endotoxins. Given that endotoxins can pose a risk for human health, the NF-κB assay is a valuable inclusion in bioanalytical test batteries used for water quality monitoring.