Pharmaceutical uptake kinetics in rainbow trout: In situ bioaccumulation in an effluent-dominated river influenced by snowmelt.

Whether seasonal instream flow dynamics influence bioaccumulation of pharmaceuticals by fish is not well understood, specifically for urban lotic systems in semi-arid regions when flows are influenced by snowmelt. We examined uptake of select pharmaceuticals in rainbow trout (Oncorhynchus mykiss) caged in situ upstream and at incremental distances downstream (0.1, 1.4, 13 miles) from a municipal effluent discharge to East Canyon Creek in Park City, Utah, USA during summer and fall of 2018. Fish were sampled over 7-d to examine if uptake occurred, and to define uptake kinetics. Water and fish tissues were analyzed via isotope dilution liquid chromatography tandem mass spectrometry. Several pharmaceuticals were consistently detected in water, fish tissue and plasma, including carbamazepine, diphenhydramine, diltiazem, and fluoxetine. Pharmaceutical levels in water ranged up to 151 ng/L for carbamazepine, whereas the effluent tracer sucralose was consistently observed at low µg/L levels. During both summer and fall experiments at each of three downstream locations from effluent discharge, rainbow trout rapidly accumulated these pharmaceuticals; tissue levels reached steady state conditions within 24-96 h. Spatial and temporal differences for pharmaceutical levels in rainbow trout directly corresponded with surface water exposure concentrations, and uptake kinetics for individual pharmaceuticals did not vary among sites or seasons. Such observations are consistent with recent laboratory bioconcentration studies, which collectively indicate inhalational exposure from water governs rapid accumulation of ionizable base pharmaceuticals by fish in inland surface waters.

Periphyton, bivalves and fish differentially accumulate select pharmaceuticals in effluent-dependent stream mesocosms.

Pharmaceuticals and other ionizable contaminants from municipal wastewater treatment plant effluent can bioaccumulate in fish, particularly in effluent dominated and dependent systems in semi-arid and arid regions. However, invertebrate bioaccumulation of these compounds has been less studied. Using municipal wastewater effluent as source water in outdoor stream mesocosms to simulate effluent-dependent lotic systems, we examined bioaccumulation of several widely-used pharmaceuticals including acetaminophen (nonsteroidal anti-inflamatory), caffeine (stimulant), carbamazepine (anti-epileptic), diltiazem (calcium channel blocker), diphenhydramine (anti-histamine), fluoxetine (anti-depressant), norfluoxetine (anti-depressant metabolite), and sertraline (anti-depressant) in freshwater clams (Corbicula fluminea), periphyton and stoneroller minnows (Campostoma anomalum), a commonly studied grazer in stream ecology, during a replicated outdoor stream mesocosm study at the Baylor Experimental Aquatic Research facility. Target analytes were determined in tissues, source effluent and stream water by isotope dilution LC-MS/MS. After an 8-day uptake period, clams accumulated a number of pharmaceuticals, including acetaminophen, carbamazepine, diltiazem, diphenhydramine, fluoxetine, norfluoxetine and sertraline with maximum concentrations reaching low µg/kg. We observed uptake rates in clams for acetaminophen at 2.8 µg/kg per day, followed by diphenhydramine (1.2 µg/kg per day) and carbamazepine (1.1 µg/kg per day). Caffeine, carbamazepine, diltiazem and diphenhydramine were measured in periphyton. Diphenhydramine was the only compound detected in all matrices, where bioaccumulation factors (BAFs) were elevated in bivalves (1631 ± 589 L/kg), compared to stoneroller minnows (247 ± 84 L/kg) and periphyton (315 ± 116 L/kg). Such BAF variability across multiple biological matrices highlight the need to understand bioaccumulation differences for ionizable contaminants among freshwater biota, including threatened and endangered species (e.g., unionids), commercially important bivalves (e.g., estuarine and marine bivalves), and fish.

Influence of salinity and pH on bioconcentration of ionizable pharmaceuticals by the gulf killifish, Fundulus grandis.

Estuaries routinely receive discharges of contaminants of emerging concern from urban regions. Within these dynamic estuarine systems, salinity and pH can vary across spatial and temporal scales. Our previous research identified bioaccumulation of the calcium channel blocker diltiazem and the antihistamine diphenhydramine in several species of fish residing in multiple urban estuaries along the Gulf of Mexico in Texas, where field-measured observations of diltiazem in fish plasma exceeded human therapeutic plasma doses. However, there remains a limited understanding of pharmaceutical bioaccumulation in estuarine environments. Here, we examined the influence of pH and salinity on bioconcentration of three pharmaceuticals in the Gulf killifish, Fundulus grandis. F. grandis were exposed to low levels of the ionizable pharmaceuticals carbamazepine, diltiazem, and diphenhydramine at two salinities (5 ppt, 20 ppt) and two pH levels (6.7, 8.3). pH influenced bioconcentration of select weak base pharmaceuticals, while salinity did not, suggesting that intestinal uptake via drinking does not appear to be a major exposure route of these pharmaceuticals in killifish. Compared to our previous pH dependent uptake observations with diphenhydramine in the fathead minnow model, killifish apparent volume of distribution values were markedly lower than fatheads, though killifish bioconcentration factors were similar at high pH and four fold higher at low pH than freshwater fish. Advancing an understanding of environmental gradient influences on pharmacokinetics among fish is necessary to improve bioaccumulation assessments and interpretation of toxicological observations for ionizable contaminants.

Spatio-temporal bioaccumulation and trophic transfer of ionizable pharmaceuticals in a semi-arid urban river influenced by snowmelt.

Bioaccumulation of pharmaceuticals in aquatic organisms is increasingly reported in the peer-reviewed literature. However, seasonal instream dynamics including occurrence and bioaccumulation across trophic positions are rarely studied, particularly in semiarid streams with flows influenced by seasonal snowmelt and municipal effluent discharges. Thus, we selected East Canyon Creek in Park City, Utah, USA to examine spatio-temporal bioaccumulation of select ionizable pharmaceuticals across trophic positions using trophic magnification factors calculated at incremental distances (0.15, 1.4, 13 miles) downstream from a municipal effluent discharge during spring (May), Summer (August), and fall (October). Nine target analytes were detected in all species during all sampling events. Trophic dilution was consistently observed for amitriptyline, caffeine, diphenhydramine, diltiazem, fluoxetine, and sertraline, regardless of seasonal instream flows or distance from effluent discharge. Calculated TMFs ranged from 0.01-0.71 with negative slopes observed for all regressions of chemical residue in tissue and trophic position. We further presents the first empirical investigation of normalizing pharmaceutical concentrations to lipid, phospholipid or protein fractions using pair matched fish samples. Empirical results identify that normalization of ionizable pharmaceutical residues in aquatic tissues to neutral lipids, polar lipids, or the total protein fraction is inappropriate, though bioaccumulation studies examining influences of internal partitioning (e.g., plasma proteins) are needed.