A catchment-based study of endocrine disruption in surface waters: multivariate evaluation of the health of a sentinel fish species exposed to sewage treatment works effluent

Summary of the results in the context of EDCAT 5 project aims: 1. By comparing appropriate biomarkers in fish sampled from STW-impacted sites and control sites during the pre-remediation period, to determine whether there was evidence for any effects that might be attributed to the presence of estrogenic (or androgenic, or anti-androgenic/-estrogenic) endocrine disrupting chemicals in the former. This aim was addressed by measuring concentrations of the estrogen-dependent yolk protein precursor vitellogenin, and the androgen-dependent nest glue spiggin in male and female sticklebacks. In addition histological examination of the gonadal structure of fish captured at the impacted and non-impacted sites was employed to seek evidence of overt alterations in reproductive physiology of the fish. For a subset of matched samples from the two rivers, the relative induction of hepatic choriogenin mRNA, a biomarker of estrogen exposure, was measured. Conclusions: Chemistry data provided by EDCAT3&4 showed that estrogenicity of the effluent was low prior to remediation and lower still following installation of the GAC plant. No evidence of overt estrogenic effects was detected in male sticklebacks in the Ray, VTG and ChG levels were similar in males from both rivers. Nor was there any evidence of alterations in spiggin concentrations in the kidneys of males from the Ray compared to the Ock. However, VTG concentrations in female sticklebacks from the Ray were increased following the STW upgrade as were hepatic ChG transcript levels, and kidney spiggin concentrations. No changes in these elements of the reproductive system were observed in females from the Ock across the same time periods. Chemical analysis of the effluent indicated that prior to installation of the GAC plant substantial concentrations of anti-androgenic chemicals were present, together with a wide range of other organics. Concentrations of these were much reduced following the plant upgrade. It is reasonable to suppose that the changes observed in the female reproductive endocrine system following the upgrade were related to the removal of some or all of this complex mixture of chemicals. The absence of effects in males may be related to the balance between exogenous and endogenous signals, or to the specificity of effects exerted by the chemicals present. No intersex fish were detected from either river. A significant bias in favour of females was detected in the stickleback populations in both rivers suggesting a factor associated with life-history of the fish, rather than contaminant burden, was responsible. 2. By comparing appropriate biomarkers in fish sampled from STW-impacted sites and control sites during the pre-remediation period, to determine whether there was evidence for any effects that might be attributed to the presence of “conventionally” toxic chemicals. This was addressed by measurement of the activity of a key Phase I transforming enzyme in the liver of fish, either using direct enzymatic assay (EROD) or by quantifying the levels of expression of the corresponding gene (CYP1A). Conclusions: EROD activity was significantly greater in fish from the Ray than the Ock in two samples collected prior to the installation of the GAC plant (2006, 2007) and this likely reflects the differential contaminant loading in the two rivers. A single sample following the commissioning of the GAC plant (2008) indicated that EROD activity had increased among fish from the Ock while that in fish from the Ray remained unchanged. While a delayed recovery of this biomarker in fish from the Ray may be expected depending on the route of exposure (direct via water or indirect via contaminated food) the reasons for elevated EROD activity in fish from the Ock/Childrey Brook are not immediately evident. Provision of a full data set for Cyp1A expression awaits the repeat of the assay. When this is complete the factors underlying the EROD findings may become clear. 3. To determine whether the adaptive capacity and energetic status of fish varied between the STW-impacted and non-impacted sites. This was addressed by measurement of indicators of stress (whole-body corticosteroid levels), metabolic status (whole-body glucose levels) and anabolic activity (RNA:DNA ratios). Conclusions: The data provide no evidence that the stress response of fish captured in the Ray prior to installation of the GAC plant was modified by exposure to the effluent. However, large variations in whole-body corticosteroid and glucose concentrations in fish from both rivers, with clear trends over time, were closely linked to perturbations in the river flow regime. Whether there was interaction between environmental and chemical factors in determining corticosteroid and glucose status is difficult to discern but it seems likely that variation in these indicators of the stress axis was driven primarily by environmental factors. The RNA:DNA ratios were closely linked with seasonal change in temperature and closely matched observed patterns of weight and length gain in stickleback populations in the two rivers. The longer growth period enjoyed by fish in the Ray was clearly evident. For both rivers, mean anabolic activity was greater during 2008 than 2007 and it seems likely that this is related to adverse effects associated with the periods of extreme flow change observed on both rivers in 2007. 4. To assess whether there were differences in population size and structure between STW-impacted and non-impacted sites. This was addressed by comparison of key somatic measures, in particular frequency distributions for fork length. Conclusions: Because of the extreme patchiness of the distribution of stickleback populations in both rivers accurate abundance estimates were not obtained. However, the catch per unit effort across the life of the project was similar for both rivers. While population size, and age structure (both rivers hosted annual populations), appeared to be similar fish in the Ray were overall larger than those from the Ock, and spawned earlier. The differences in growth and timing of spawning between the rivers were likely to have been associated with the Rodbourne STW effluent. Downstream of the discharge on the Ray water temperatures were consistently 2 – 3oC above those of the Ock. This temperature difference, in combination with the introduction of additional nutrients into the river which is likely to have affected the availaibility of food, probably accounts for the different growth profile among the sticklebacks in the two rivers. However, over and above this difference, there was a significant increase in size of sticklebacks in the Ray between the matched pre- and post-remediation periods in the Ray while no change in size of the fish in the Ock occurred during the same period. Similarly, the RNA:DNA ratio was higher in fish from the Ock during 2007 but greater in fish from the Ray during 2008. Taken together, these observations suggest that there was an improvement in the status of the fish in the Ray following the commissioning of the GAC plant, while the population in the Ock remained relatively stable. It is reasonable to suppose that this may be linked to the reduction of the chemical load entering the Ray at Rodbourne following the installation of the GAC plant. The Ray is “cleaner” now than was the case prior to remediation but remains nutrient rich and several degrees warmer than the Ock, this combination of factors providing fish in the Ray with greater scope for growth relative to populations in the Ock

The distribution of Polychlorinated Biphenyls (PCBs) in the River Thames Catchment under the scenarios of climate change

Measurements have shown low levels of PCBs in water but relatively high concentrations in the resident fish of the River Thames (UK). To better understand the distribution and behaviour of PCBs in the Thames river basin and their potential risks, a level III fugacity model was applied to selected PCB congeners (PCB 52, PCB 118 and PCB 153). The modelling results indicated that fish and sediments represent environmental compartments with the highest PCB concentrations; but the greatest mass of PCBs (over 70%) is likely to remain in the soil. As emissions decline, soil could then act as a significant secondary source of PCBs with the river bed-sediment functioning as a long-term reservoir of PCBs. The predicted changes in temperature and rainfall forecast in the UK Climate Projections 2009 (UKCP09) over the next 80 years had only a modest influence on PCB fate in the model. The most significant result was a tendency for climate change to enhance the evaporation of PCBs from soil to air in the Thames catchment. (C) 2015 Elsevier B.V. All rights reserved

The influence of exposure and physiology on microplastic ingestion by the freshwater fish Rutilus rutilus (roach) in the River Thames, UK

Microplastics are widespread throughout aquatic environments. However, there is currently insufficient understanding of the factors influencing ingestion of microplastics by organisms, especially higher predators such as fish. In this study we link ingestion of microplastics by the roach Rutilus rutilus, within the non-tidal part of the River Thames, to exposure and physiological factors. Microplastics were found within the gut contents of roach from six out of seven sampling sites. Of sampled fish, 33% contained at least one microplastic particle. The majority of particles were fibres (75%), with fragments and films also seen (22.7% and 2.3% respectively). Polymers identified were polyethylene, polypropylene and polyester, in addition to a synthetic dye. The maximum number of ingested microplastic particles for individual fish was strongly correlated to exposure (based on distance from the source of the river). Additionally, at a given exposure, the size of fish correlated with the actual quantity of microplastics in the gut. Larger (mainly female) fish were more likely to ingest the maximum possible number of particles than smaller (mainly male) fish. This study is the first to show microplastic ingestion within freshwater fish in the UK and provides valuable new evidence of the factors influencing ingestion that can be used to inform future studies on exposure and hazard of microplastics to fish

Size dependence of silver nanoparticle removal in a wastewater treatment plant mesocosm measured by FAST single particle ICP-MS

The quantities of engineered nanoparticles (NP) released to the environment are often influenced by their fate in waste water treatment plants (WWTP). Here, 40 nm silver NP (AgNP) were spiked into a mesocosm simulating the process used at a major municipal WWTP. The evolution of the mass distributions and number concentrations were followed by fast acquisition speed technique single particle inductively coupled mass spectrometry (FAST spICP-MS) using a high-resolution ICP-MS. It was thus possible to detect smaller Ag containing NP than hitherto possible in similar studies. These small particles (ca. 5-10 nm in corresponding metallic Ag equivalent spherical diameter) were possibly dissolved Ag+ precipitated as Ag2S particles. They were detected immediately upon spiking and were stable with respect to aggregation and thus much less removed by the WWTP process compared to the 40 nm AgNP. The results also suggested that any transformation of the latter AgNP occurred without dissolution. Most of these larger AgNP were probably removed by aggregation with large floc particles and subsequent sedimentation with the suspended particulate matter in the simulated WWTP process. The results have implications for differentiating the fate of nanoparticles as a function of size and demonstrate how spICP-MS can reveal such size-dependent fate dynamics

Which metal represents the greatest risk to freshwater ecosystem in Bohai Region of China?

Metals discharged from industrial effluents, agricultural wastewater, and sewage runoff by rapid urbanization are of concern as contaminants of freshwater ecosystem because of their persistence and high toxicity to aquatic organisms. This study attempted to identify which metal posed the greatest risk to freshwater ecosystem in the Bohai Region, China. The metals arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), zinc (Zn), lead (Pb), nickel (Ni), mercury (Hg), iron (Fe), and manganese (Mn) were compared against norfloxacin and gamma-hexachlorocyclohexane (lindane). By comparing the median reported environmental and ecotoxicity concentrations, it showed that Cu, Fe, Zn, Mn, and Cr were the top five metals of concern. Of these, Cu was deemed to represent the highest risk and Hg the lowest risk. The risks for all metals were higher than those for norfloxacin and lindane. Almost all the metals except Hg had water concentrations that exceeded levels where ecotoxicity effects had been recorded in the literature. A comparison with the measurements across the UK rivers suggested that all metals examined had water concentrations about 5- to 10- fold higher than the UK median values except for Cu, Fe, Cd, and Pb. The Fuyang River, a tributary of the Haihe River Basin, seemed to be the location with the highest metal concentrations. However, comparing the post- 2010 period to 2000–2009, concentrations of all the metals had fallen except for Fe and Mn, so risks have decreased over the last 7 yr with the greatest improvements for Cd and Pb. While metals still pose high risks to freshwater ecosystem in this region, there is encouragement that some control measures are taken into effect

The different fate of antibiotics in the Thames River, UK, and the Katsura River, Japan

Little is known about the mechanisms influencing the differences in attenuation of antibiotics between rivers. In this study, the natural attenuation of four antibiotics (azithromycin, clarithromycin, sulfapyridine, and sulfamethoxazole) during transport along the Thames River, UK, over a distance of 8.3 km, and the Katsura River, Japan, over a distance of 7.6 km was compared. To assist interpretation of the field data, the individual degradation and sorption characteristics of the antibiotics were estimated by laboratory experiments using surface water or sediment taken from the same rivers. Azithromycin, clarithromycin, and sulfapyridine were attenuated by 92, 48, and 11% in the Thames River stretch. The first-order decay constants of azithromycin and sulfapyridine were similar to those in the Katsura River, while that of clarithromycin was 4.4 times higher. For sulfamethoxazole, the attenuation was limited in both rivers. Loss of sulfapyridine was attributed to both direct and indirect photolysis in the Thames River, but to only direct photolysis in the Katsura River. Loss of azithromycin and clarithromycin was attributed to sorption to sediment in both rivers. The probable explanation behind the difference in loss rates of clarithromycin between the two rivers was considered to be sediment sorption capacity

Linking changes in antibiotic effluent concentrations to flow, removal and consumption in four different UK sewage treatment plants over four years

The arrival and discharge of seven antibiotics were monitored at two trickling filter sewage treatment plants of 6,000 and 11,000 population equivalents (PE) and two activated sludge plants of 33,000 and 162,000 PE in Southern England. The investigation consisted of 24 h composite samples taken on two separate days every summer from 2012 to 2015 and in the winter of 2015 (January) from influent and effluent. The average influent concentrations generally matched predictions based on England-wide prescription data for trimethoprim, sulfamethoxazole, azithromycin, oxytetracycline and levofloxacin (within 3-fold), but were 3-10 times less for clarithromycin, whilst tetracycline influent concentrations were 5-17 times greater than expected. Over the four years, effluent concentrations at a single sewage plant varied by up to 16-fold for clarithromycin, 10-fold for levofloxacin and sulfamethoxazole, 7-fold for oxytetracycline, 6-fold for tetracycline, 4-fold for azithromycin and 3-fold for trimethoprim. The study attempted to identify the principal reasons for this variation in effluent concentration. By measuring carbamazepine and using it as a conservative indicator of transport through the treatment process, it was found that flow and hence concentration could alter by up to 5-fold. Measuring influent and effluent concentrations allowed assessments to be made of removal efficiency. In the two activated sludge plants, antibiotic removal rates were similar for the tested antibiotics but could vary by several-fold at the trickling filter plants. However, for clarithromycin and levofloxacin the variations in effluent concentration were above that which could be explained by either flow and/or removal alone so here year on year changes in consumption are likely to have played a role

Assessing the population equivalent and performance of wastewater treatment through the ratios of pharmaceuticals and personal care products present in a river basin: application to the River Thames basin, UK

The quality of surface waters in lowland rivers is largely dependent on the efficiency of wastewater treatment. Even in the developed countries, there have been difficulties in evaluating the effectiveness of wastewater management and the proportion of wastewater content (WWC) in the river, as well as in estimating the contributing human population. This study aimed to develop a wastewater quality and quantity assessment based on the occurrence of pharmaceuticals in the receiving waters. A survey of 53 pharmaceuticals in 324 samples (river water and influent and effluent of sewage (wastewater) treatment plants) was carried out in southern England in the River Thames catchment over four years. Carbamazepine was selected as stable marker and from its concentration WWC in the rivers and cumulative human populations along the catchment were estimated. The estimated population had a strong relationship (R2=0.94) with that reported by the local water company. The concentration ratio of the labile marker caffeine to carbamazepine indicated the efficiency of wastewater treatment in the different treatment systems (i.e. trickling filter or activated sludge) and in the receiving waters. The ratio in some river samples revealed unexpected discharges of untreated or poorly treated wastewater, with a total concentration of the analytes (up to 20mug/L) five times higher than that in treated wastewater. Such information could be valuable to estimate the discharge or occurrence of not only non-targeted chemicals, but also pathogens within the basin

Which persistent organic pollutants in the rivers of the Bohai region of China represent the greatest risk to the local ecosystem?

Freshwater aquatic organisms can be exposed to hundreds of persistent organic pollutants (POPs) discharged by natural and anthropogenic activities. Given our limited resources it is necessary to identify, from the existing evidence, which is the greatest threat so that control measures can be targeted wisely. The focus of this study was to rank POPs according to the relative risk they represent for aquatic organisms in rivers in the Bohai Region, China. A list of 14 POPs was compiled based on the available data on their presence in these rivers and ecotoxicological data. Those that were widely detected were benzo[a]pyrene, p,p'-DDE, p,p'-DDT, endrin, fluoranthene, heptachlor, hexabromocyclododecane, hexachlorobenzene, alpha-hexachlorocyclohexane, gamma-hexachlorocyclohexane, naphthalene, perfluorooctanoic acid, perfluorooctane sulfonate and phenanthrene. Effect concentrations were compiled for Chinese relevant and standard test species and compared with river aqueous concentrations. Only bed-sediment concentrations were available so water levels were calculated based on the known local sediment organic carbon concentration and the Koc. The POPs were ranked on the ratio between the median river and median effect concentrations. Of the POPs studied, fluoranthene was ranked as the highest threat, followed by phenanthrene, naphthalene and p,p'-DDE. The risk from p,p'-DDE may be magnified due to being highly bioaccumulative. However, the greatest overlap between river concentrations and effect levels was for lindane. Overall, fish was the most sensitive species group to the risks from POPs. Hotspots with the highest concentrations and hence risk were mainly associated with watercourses draining in Tianjin, the biggest city in the Bohai Region

Quantification of pharmaceutical related biological activity in effluents from wastewater treatment plants in UK and Japan

While pharmaceuticals are now routinely detected in aquatic environments, we know little of the biological activity their presence might provoke. It is estimated that nearly 40% of all marketed pharmaceuticals are G protein-coupled receptors (GPCRs) acting pharmaceuticals. Here, we applied an in-vitro assay, called the TGFα shedding assay, to measure the biological activities of GPCRs-acting pharmaceuticals present in effluents from municipal wastewater treatment plants in the United Kingdom (UK) and Japan from 2014 to 2016. The results indicated that compounds were present in the wastewater with antagonistic activities against angiotensin (AT1), dopamine (D2), adrenergic (β1), acetylcholine (M1), and histamine (H1) receptors in both countries. The most consistent and powerful antagonistic activity was against the H1, D2, and AT1 receptors at up to microgram-antagonist-equivalent quantity/L. Chemical analysis of the same UK samples was also conducted in parallel. Comparing the results of the bioassay with the chemical analysis indicated (1) the existence of other D2 or M1 receptor antagonists besides sulpiride (D2 antagonist) or pirenzepine (M1 antagonist) in wastewater and (2) that there might be a mixture effect between agonist and antagonistic activities against β1 receptor. GPCR-acting pharmaceuticals should be paid more attention in the environmental monitoring and toxicity testing in future studies