Peter van den Hurk

The induction of biochemical changes in Daphnia magna by CuO and ZnO nanoparticles

Whilst a considerable number of studies have been reported on the acute toxicity of nanoparticles (NPs) on invertebrates such as Daphnia magna, few studies have been reported on the biochemical change (biomarkers) induction on these species by NPs, especially metal oxide NPs. The aim of this study was to investigate some biomarkers in D. magna induced by copper oxide (CuO) and zinc oxide (ZnO) NPs under controlled laboratory conditions. We exposed the 5 day old D. magna for 72 h to sublethal concentration of CuO and ZnO NPs in synthetic moderately hard water (MHW) with and without dissolved natural organic matter (NOM) and estimated the glutathione-S-transferase (GST) activity, formation of oxidized glutathione (GSSG), and amounts of thiobarbituric acid reacting substances (TBARS) and metallothionein (MT). Additionally, complementary short term dissolution studies on CuO and ZnO NPs were conducted. The results showed inactivation of GST enzyme by both metal oxide NPs. The results also showed increased production of oxidized GSH, increased generation of TBARS and increased induction of MT. In the presence of NOM, significant reduction (p<0.05) in these biochemical changes was observed. These results indicated that oxidative stress is one of the toxicity mechanisms for these metal oxide NPs. Furthermore, the results suggest that these metal oxide NPs compromise the health of D. magna, and possibly other aquatic organisms, and therefore have potential to affect ecosystem stability. The short term dissolution studies showed that the proportion of dissolved NPs is higher (1.2% and 70% of initial concentration for dissolved Cu and Zn, respectively) at low particle concentration and is lower (0.4% and 17% of initial concentration for dissolved Cu and Zn, respectively) at higher particle concentration. These results suggest that the observed toxicity may be caused by both metal oxide nanoparticles and metal ions dissociated from the nanoparticles.

A BIOMARKER APPROACH TO MEASURE BIOLOGICAL EFFECTS OF CONTAMINANT EXPOSURE IN LARGEMOUTH BASS FROM LAKE CONESTEE, SOUTH CAROLINA, USA

Sediments from Lake Conestee, a former reservoir now filled with pollutant-enriched sediments, located south of Greenville, South Carolina, USA, and other nearby reservoirs were collected and analyzed for lead and polycyclic aromatic hydrocarbons (PAHs). Hepatic ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), UDP-glucuronosyltransferase (UGT), sulfotransferase (SULT), and erythrocyte delta-aminolevulinic acid dehydratase (ALAD) were measured in largemouth bass (Micropterus salmoides) to characterize biological effects of these contaminants over three seasons. Results showed that total PAH concentrations in Lake Conestee sediments were significantly greater than the control during each season. An average 10-fold induction in EROD activity was observed at Lake Conestee compared with the control over all three seasons, indicating that PAHs present in sediment were bioavailable to fish. Significant gender effects were observed in EROD activity during the spring, in which activity in reproductively active female fish was significantly suppressed compared with the male fish. Sediments from Lake Conestee had elevated lead concentrations, but the lack of ALAD inhibition in bass indicated that lead was not biologically available. Total GST activity, UGT activity, and SULT activity were not significantly induced in fish from any of the affected sites compared with the reference site. Both EROD and UGT activities were highest during the winter, as were sediment PAH concentrations in Lake Conestee, possibly linked to seasonal resuspension events. The biomarkers measured in this study were useful as a first investigation into the biological effects of contaminant exposure, as well as in determining the bioavailability of contaminants in Lake Conestee.

Influence of carbon nanotubes on the bioavailability of fluoranthene

Concurrent with the increase in the use of carbon nanotubes (CNTs) in society is the rise of their introduction into the environment. Carbon nanotubes cause adverse effects themselves, and they have the potential to adsorb contaminants such as polycyclic aromatic hydrocarbons (PAHs). Although CNTs have a high adsorption capacity for PAHs and these contaminants can co-occur in the environment, few studies have characterized the bioavailability of CNT-adsorbed PAHs to fish. The goal of the present study was to characterize the bioavailability of fluoranthene adsorbed to suspended multiwalled-carbon nanotubes (MWNTs) in freshwater containing natural organic matter (NOM). Adsorption isotherms indicated that NOM influenced the adsorption of fluoranthene to MWNTs, although in the absence of MWNTs it did not influence the bioavailability of fluoranthene to Pimephales promelas. Pimephales promelas were exposed for 16 h in synthetic moderately hard water containing fluoranthene in the presence of different concentrations of NOM, and fluoranthene adsorbed to MWNTs in the presence of NOM. Bioavailable fluoranthene was quantified in each exposure through bile analysis using fluorescence spectrophotometry. By comparing the concentration of fluoranthene metabolites in the bile with the concentration of fluoranthene added to MWNT and NOM solutions, the relative bioavailability of fluoranthene adsorbed to MWNTs was quantified. Results indicate that approximately 60% to 90% of the fluoranthene was adsorbed to the MWNTs and that adsorbed fluoranthene was not bioavailable to P. promelas. The results also suggest that fluoranthene is not desorbed from ingested MWNT, and the bioavailable fraction is only the freely dissolved fluoranthene in the aqueous phase.

Trophic transfer of microplastics in aquatic ecosystems: Identifying critical research needs

To evaluate the process of trophic transfer of microplastics, it is important to consider various abiotic and biotic factors involved in their ingestion, egestion, bioaccumulation, and biomagnification. Toward this end, a review of the literature on microplastics has been conducted to identify factors influencing their uptake and absorption; their residence times in organisms and bioaccumulation; the physical effects of their aggregation in gastrointestinal tracts; and their potential to act as vectors for the transfer of other contaminants. Limited field evidence from higher trophic level organisms in a variety of habitats suggests that trophic transfer of microplastics may be a common phenomenon and occurs concurrently with direct ingestion. Critical research needs include standardizing methods of field characterization of microplastics, quantifying uptake and depuration rates in organisms at different trophic levels, quantifying the influence that microplastics have on the uptake and/or depuration of environmental contaminants among different trophic levels, and investigating the potential for biomagnification of microplastic-associated chemicals. More integrated approaches involving computational modeling are required to fully assess trophic transfer of microplastics. Integr Environ Assess Manag 2017;13:505-509.

Effects of an antidepressant mixture on the brain serotonin and predation behavior of hybrid striped bass

Antidepressants have been found in measurable concentrations in final treated wastewater effluent and receiving waters throughout the world. Studies have shown that these concentrations are typically not overtly toxic, but the psychotropic mode of action of these chemicals warrants examination of their behavioral effects. Exposure of hybrid striped bass to the antidepressants fluoxetine or venlafaxine alone has been shown to cause decreased brain serotonin levels and increased time to capture prey at concentrations typically 1 to 2 orders of magnitude higher than environmentally relevant concentrations. In the present study, equally effective doses of fluoxetine and venlafaxine were used to perform a mixture study, using a toxic unit approach to determine whether these antidepressants may act in an additive manner at lower concentrations. The results indicated that mixtures of these antidepressants caused decreased brain serotonin and increased time to capture prey at concentrations lower than reported in previous studies. Low concentration mixtures caused an additive effect on brain serotonin levels and time to capture prey, whereas higher concentrations were less than additive. The results were consistent with the dose addition concept, with higher concentration mixtures potentially saturating the effects on serotonin in the brain. Results from the present study indicate that antidepressants have the potential to be additive on the biochemical and individual scale, which necessitates more robust analysis of antidepressant mixtures and their potential to act together in low concentration scenarios.

Biomarker responses in sunfish species and largemouth bass from the Saluda River, South Carolina.

The upstate and Piedmont region of South Carolina is a rapidly urbanizing area as a result of a steadily growing population. This increase in population and development has the potential to negatively impact local aquatic systems like the Saluda River due to increased pollution from runoff, and effluents from industrial and wastewater treatment facilities. During the summer months of 2010, 159 fish from the Centrarchidae family (sunfish species (Lepomis) and largemouth bass - Micropterus salmoides) were collected from 13 sites along the Saluda River. A suite of biomarker assays, including ethoxyresosufin-O-deethylase, bile fluorescence, glutathione S-transferase, thiobarbituric acid reactive substances, bile estrogens, acetylcholinesterase inhibition, metallothionein and tissue metal levels were applied to investigate the impacts of diminished water quality on fish health. Results indicate that fish from the Saluda River are responding to contamination in a site specific manner, with up to four significant biomarker responses in the most impacted sites. Sampling sites in the lower portion of the Saluda watershed are less impacted by pollution than the upper and central sections. The observed biomarker responses can be explained by the proximity of urban areas, point sources and general land use, and demonstrate the applicability of biomarkers in environmental biomonitoring programs.

Xenoestrogen Exposure and Effects in Bluegill from the Reedy River, South Carolina, USA

Water quality in the Reedy River basin of Greenville, South Carolina, has been impacted by diverse and highly urbanized land uses. It has been demonstrated that urban runoff and point sources, such as effluent from wastewater treatment facilities, introduce organic pollutants and potentially endocrine disrupting compounds (EDCs) into the watershed. The objective of this study was to investigate the potential toxicological effects of EDCs that may be present in the Reedy River watershed using a set of biomarkers measured in indigenous fish to characterize the exposure and biological effects of these contaminants. Bluegill (Lepomis macrochirus) were collected during three different sampling seasons (spring, summer, and fall) from several sites along the length of the Reedy River and from an unimpacted site at Lake Robinson. Fish were analyzed for xenoestrogenic exposure (estrogenic effect of bile extracts) and effects (vitellogenin production in juvenile fish), which were compared to the hepatosomatic index as a general health parameter. Samples downstream of Greenville, especially downstream of the wastewater treatment facilities, were found to have significantly higher levels of estrogenic activity in bile extracts, which correlated well with elevated plasma vitellogenin concentrations relative to the specimens collected in reference sites. The results provide evidence that bluegill in the Reedy River were exposed to elevated concentrations of xenoestrogenic compounds and that these xenoestrogens were bioavailable, resulting in biological effects.

Seafood substitutions obscure patterns of mercury contamination in Patagonian toothfish (Dissostichus eleginoides) or "Chilean sea bass".

Seafood mislabeling distorts the true abundance of fish in the sea, defrauds consumers, and can also cause unwanted exposure to harmful pollutants. By combining genetic data with analyses of total mercury content, we have investigated how species substitutions and fishery-stock substitutions obscure mercury contamination in Patagonian toothfish (Dissostichus eleginoides), also known as “Chilean sea bass”. Patagonian toothfish show wide variation in mercury concentrations such that consumers may be exposed to either acceptable or unacceptable levels of mercury depending on the geographic origins of the fish and the allowable limits of different countries. Most notably, stocks of Patagonian toothfish in Chile accumulate significantly more mercury than stocks closer to the South Pole, including the South Georgia/Shag Rocks stock, a fishery certified by the Marine Stewardship Council (MSC) as sustainably fished. Consistent with the documented geography of mercury contamination, our analysis showed that, on average, retail fish labeled as MSC-certified Patagonian toothfish had only half the mercury of uncertified fish. However, consideration of genetic data that were informative about seafood substitutions revealed a complex pattern of contamination hidden from consumers: species substitutions artificially inflated the expected difference in mercury levels between MSC-certified and uncertified fish whereas fishery stock substitutions artificially reduced the expected difference in mercury content between MSC-certified and uncertified fish that were actually D. eleginoides. Among MSC-certified fish that were actually D. eleginoides, several with exogenous mtDNA haplotypes (i.e., not known from the certified fishery) had mercury concentrations on par with uncertified fish from Chile. Overall, our analysis of mercury was consistent with inferences from the genetic data about the geographic origins of the fish, demonstrated the potential negative impact of seafood mislabeling on unwanted mercury exposure for consumers, and showed that fishery-stock substitutions may expose consumers to significantly greater mercury concentrations in retail-acquired fish than species substitutions.

Assessment of heavy metal and PAH exposure in largemouth bass (Micropterus salmoides) in the Reedy River watershed, South Carolina, USA: A multi‐season assessment of metallothionein and bile fluorescence

Biomarkers can be used as tools to help determine ecological exposure in watershed assessments. In the present study, metallothionein and fixed wavelength bile fluorescence for two-, four-, and five-ring hydrocarbons were used as biomarkers of exposure in largemouth bass (Micropterus salmoides) in the Reedy River watershed located in South Carolina, USA. Fish were sampled from three impoundments and a reference site over three seasons in the same year. Biomarker endpoints were compared to chemical concentrations at each site during each season. Results indicated that despite elevated concentrations of hydrocarbons being present in sediments, the hydrocarbons did not appear to be bioavailable based on bile fluorescence analysis. Bile fluorescence analysis also indicated that the hydrocarbons detected in this watershed were likely of petrogenic origin. Significantly elevated sediment concentrations of Cd, Cr, Cu, Pb, Ni, Ag, Zn, and metallothionein were found in one impoundment, Lake Conestee, compared with the reference site, indicating both the presence and bioavailability of these metals. Seasonal variability of bile fluorescence was limited; however, metallothionein showed elevated concentrations in the spring and summer compared with fall.

Phylogenetic signals in detoxification pathways in Cyprinid and Centrarchid species in relation to sensitivity to environmental pollutants

Observations in a previous study on biomarker responses in fish collected from urban creeks in Greenville, SC, indicated that there might be considerable differences in the expression of biotransformation enzymes in chub and sunfish species. To further investigate these species differences a dosing experiment was performed in which bluehead and creek chub (Nocomis leptocephalus and Semotilus atromaculatus), and redbreast sunfish, pumpkinseed, and bluegill (Lepomis auritus, L. gibbosus, and L. macrochirus) were injected with benzo[a]pyrene (BaP) as a model compound for common pollutants in urban creeks. Fish were injected with BaP doses of 0, 25 and 50mg/kg, and after 3days BaP metabolites in bile, and enzymatic activities of cytochrome P450-1A (CYP1A), UDP-glucuronosyltransferase (UGT) and glutathione S-transferase (GST) were measured. CYP1A activity was significantly increased after BaP dosing in both species groups, but chubs had significantly lower levels than were observed in the dosed sunfish. The UGT activity in unexposed animals was comparable in both species groups, and significantly increased in both groups as a result of BaP dosage. Finally, GST activity was significantly higher in chubs, but did not change in either species group as a result of BaP exposure. There were no significant differences between species within each species group, and the results confirmed that unexposed chubs have much lower CYP1A activity, but a much higher GST activity than unexposed sunfish. The metabolized BaP was excreted in both species groups, but at the time of sampling there were no differences in the amount of BaP metabolites in the bile of dosed animals. The differences in baseline enzyme activity and induction capacity between both species groups are an example of phylogenetically determined differences between fish families, and may explain why chubs are in general more sensitive to exposure to environmental pollutants than sunfish. This conclusion was corroborated by the observation that the highest BaP dose of 50mg/kg was close to the apparent LC50 for chub, while no mortality was observed in the sunfish at this dose.