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

ABSTRACT: 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).

ORGANISM(S): Danio rerio

PROVIDER: GSE152131 | GEO | 2020/11/30

REPOSITORIES: GEO

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Project description:Three surface waters in Gainesville, Florida were used in a 48 hour whole effluents exposure to assess gene expression profiles of male fathead minnow liver. Microarray analysis was used to determine changes in gene expression of exposed fish to waters from a site downstream of a wastewater treatment plant (streamwater), a wastewater treatment plant (wastewater), and a lake (stormwater). Differences in gene expression between fish exposed to collected waters and controls were observed. Number of altered genes and biological processes were 1028 and 18 for stormwater; 787 and 19 for streamwater; and: 575 and 12 for wastewater. In general, the effects observed in all exposed fish were related with fatty acid metabolism, DNA repair, oxidation-reduction process, cell wall catabolic process and apoptosis. All exposed fish showed altered expression of genes related with DNA damage repair. In particular fish exposed to stormwater and streamwater showed downregulation of several key intermediates transcripts of cholesterol. The presence and environmental persistence of perfluorinated chemicals (PFCs) in these waters, the resemblance in known effects on transcripts with those found in this study, suggest that the set of genes differentially regulated in fathead minnows after 48 hours of exposure may be attributed to exposure to PFCs. Three surface water sites were chosen for effluent collection in Gainesville, Florida: A lake (stormwater), surface water downstream of a wastewater treatment plant (streamwater), and a wastewater treatment plant effluent used for landscaping irrigation (wastewater). Water from each site was collected two days prior to the fish exposure experiment using Chemfluor ® tubing and a 120 liters steel barrels coated with polyester resin (gel coat) to avoid cross-contamination. Three barrels for each effluent were collected during day 1. Water from the barrel was transported to the laboratory and pumped into four fiberglass cylinders in the aquatic toxicology facility. Water from each cylinder was then pumped into four replicate aquariums per treatment and kept for 1 day without fish (pre-treatment). On day 2, four male fathead minnows from a common tank were transferred to each replicate aquarium and kept for 48 hours, with one 75% water change after first 24 hours. The exposure system consisted of 40 L glass aquaria. Each exposure was conducted in quadruplicate and each aquarium contained the four male fish in 25 L of treatment water . The water used in the control treatment was carbon filtered, dechlorinated tap water. The positions of the treatment tanks were randomized and test initiation times were staggered to ensure an exposure/sampling interval of 48 h. The fish were not fed during the experiment. The temperature range of the water was 24-26 °C with a photoperiod of 16 h light: 8 h dark. Liver was isolate from 4 males indviduals for each treatment except for control group (3 individuals).

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

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