An Investigation into the Biological Effects of Indirect Potable Reuse Water Using Zebrafish Embryos

In the search for sustainable drinking water, many countries are weighing up the benefits of advanced treatment technologies as a proactive measure to assist with the transformation of treated wastewater into a source of water used for the production of potable water. We investigated the biological effects along a pilot plant with an advanced water treatment process, using zebrafish embryos at different stages of development. The study took an innovative approach, comparing phenotypic observations with whole genome responses. This enabled us to keep an open mind about which chemicals might be influencing the biological activity. There was no evidence of acute toxicity at any stage of treatment, but distinctive abnormalities – skeletal, cardiovascular and pigmentation – occurred in a small proportion of embryos along the treatment process, and in a tap water, that were not detected in the aquarium water control. Reverse osmosis (RO) reduced the concentration of measured chemical contaminants in the water the most, whilst eliminating the occurrence of abnormalities detected in the fish. In contrast, advanced oxidation appeared to reverse the benefits of RO treatment by increasing the frequency of teratogenic and sub-lethal abnormalities seen in embryos. Genomic analysis found alterations to the retinoid system, which was consistent with the teratogenic abnormalities observed. In addition, we found evidence of changes to metabolic pathways, including tryptophan metabolism associated with the production of melatonin required for the control of normal circadian rhythms. Although we cannot extrapolate these preliminary findings in zebrafish embryos to human or environmental health, we show that underexplored forms of biological activity (that existing Test Guidelines are not designed to capture) occur in treated wastewater effluent, and/or may be created depending on the type of advanced treatment process used. Although the identity of the culprit chemicals are unknown at this time, our innovative approach highlights the need for more research into the effects of chemicals on the retinoid system (and metabolism). Analysis of water from a indirect potable reuse water plant. Stages of the water purification were tested on effects on Zebrafish embryos. These stages included AOP-1, AOP-2, MF, RAW, RO, Tap, a separate water control and the Final effluent. Exposures were performed over time (8,12,16,24,36, and 48h). In total 132 samples were generated.