Do food and stress biomarkers work for wastewater-based epidemiology? A critical evaluation.

Dietary characteristics and oxidative stress are closely linked to the wellbeing of individuals. In recent years, various urinary biomarkers of food and oxidative stress have been proposed for use in wastewater-based epidemiology (WBE), in efforts to objectively monitor the food consumed and the oxidative stress experienced by individuals in a wastewater catchment. However, it is not clear whether such biomarkers are suitable for wastewater-based epidemiology. This study presents a suite of 30 urinary food and oxidative stress biomarkers and evaluates their applicability for WBE studies. This includes 22 biomarkers which were not previously considered for WBE studies. Daily per capita loads of biomarkers were measured from 57 wastewater influent samples from nine Australian catchments. Stability of biomarkers were assessed using laboratory scale sewer reactors. Biomarkers of consumption of vitamin B2, vitamin B3 and fibre, as well as a component of citrus had per capita loads in line with reported literature values despite susceptibility of degradation in sewer reactors. Consumption biomarkers of red meat, fish, fruit, other vitamins and biomarkers of stress had per capita values inconsistent with literature findings, and/or degraded rapidly in sewer reactors, indicating that they are unsuitable for use as WBE biomarkers in the traditional quantitative sense. This study serves to communicate the suitability of food and oxidative stress biomarkers for future WBE research.

A macroalgal germling bioassay to assess biocide concentrations in marine waters.

A bioassay method using the early life stages (germlings) of macroalgae was developed to detect toxicity of anti-fouling paint biocides. A laboratory based bioassay using Ulva intestinalis and Fucus spiralis germlings was performed with 4 common anti-fouling biocides (tributyltin (TBT), Irgarol 1051, Diuron and zinc sulphate), over a range of environmentally relevant concentrations (0.0033-10 µg l(-1)). Comparison between the two species showed that germlings of U. intestinalis were better adapted for in-situ monitoring, as germlings of F. spiralis appeared to be too robust to display sufficient growth differences. The response of U. intestinalis germling growth appeared to reflect environmental biocide concentrations. Overall the developed method showed potential for the assessment of the sub-lethal effects of anti-fouling biocides on the early developmental stages of U. intestinalis.

The binding of phenanthrene to engineered silver and gold nanoparticles.

The steadily rising production and use of engineered nanoparticles (ENP) leads to their entry into the aquatic environment. In addition to the various adverse effects that have been seen for different organisms, ENP are suspected to influence the transport, bioavailability and toxic properties of a range of environmental contaminants that may adsorb to their surface. In this study, the binding properties of the polycyclic aromatic hydrocarbon phenanthrene to stabilized silver and gold ENP were investigated using a novel mass balance based single-equilibrium approach. Only citrate coated gold ENP (AuNP(CIT)) were found to bind phenanthrene. No binding was observed for polyvinylpyrolidone coated silver ENP (AgNP(PVP)) nor citrate coated silver ENP (AgNP(CIT)) suggesting that the properties of the core material have a major influence on binding reactions. A binding coefficient K(b) was defined as the ratio between the concentration of phenanthrene associated to the AuNP(CIT) and that freely dissolved in the exposure medium. Temperature was not seen to significantly influence K(b) within an environmentally relevant range (4-25 °C). The presence of methanol significantly reduced or prevented the formation of the AuNP(CIT)-phenathrene complex. Results suggest that the binding is a low energy physio-sorption, likely associated to a partial displacement or specific arrangement of the citrate capping on the gold core.

The environmental fate and effects of antifouling paint biocides.

Antifouling (AF) biocides are the active ingredients in AF paints that prevent the settlement, adhesion and growth of organisms to a painted surface. A wide range of chemicals are used as AF biocides, which have very different physico-chemical properties and therefore differing environmental fates, behaviour and effects. Copper has been used as an antifoulant for centuries and extensive research has been performed to understand how copper speciation influences bioavailability and toxicity. For biocides that have been widely used over a number of decades, for example Irgarol 1051 and diuron, there are a large amount of environmental data in the public domain, including for their respective metabolites, that allows their environmental safety and potential risk to the environment to be assessed. For other biocides such as dichlofluanid, DCOIT (SeaNine 211) and zinc/copper pyrithione, there is a good understanding of their fate and effects. However, few monitoring studies have been performed and not so much is known about the fate and effects of their metabolites. There are also new or candidate biocides such as triphenylborane pyridine, Econea, capsaicin and medetomidine for which there is very little information in the public domain. This review provides an overview of the environmental fate and occurrence data that are in the public domain for AF biocides and provides some insight into the effects of these compounds on non-target organisms.

Effect-directed identification of naphthenic acids as important in vitro xeno-estrogens and anti-androgens in North sea offshore produced water discharges.

Produced water from offshore oil production platforms represents the largest direct discharge of effluent into the offshore environment. Produced water effluents contain a complex mixture of substances which are known to bind to the estrogen receptor (ER) and antagonize the androgen receptor (AR). Short-chain petrogenic alkylphenols have been identified as responsible for around 35% of the ER agonist activity measured in vitro while the compounds responsible for antagonizing the androgen receptor are unknown. For the first time we report that petrogenic naphthenic acids are weak ER agonists that account for much of the 65% of the "unknown" ER agonist potency in North Sea produced waters while also disrupting the binding of AR agonists to the AR ligand receptor. We also report other known petrogenic components such as polycyclic aromatic hydrocarbons (PAHs) and alkylphenols as environmental AR antagonists. Our investigation shows that these petrogenic components are responsible for the majority of the ER and AR receptor mediated activity in produced waters. This hypothesis is supported by data from an effects-directed analysis of produced water using normal-phase high-performance liquid chromatography (HPLC) fractionation in combination with the yeast estrogen and androgen assays as well as androgen receptor binding assays of commercially available mixtures of naphthenic acids.

Ecotoxicity of the degradation products of triphenylborane pyridine (TPBP) antifouling agent.

Triphenylborane pyridine (TPBP) is an alternative to organotin antifouling compounds. This work aimed to identify the unknown Peak #1, and to evaluate the ecotoxicity of TPBP and its degradation products. Peak #1 was produced from TPBP dissolved in acetonitrile under UV-A photolysis using a high-pressure mercury lamp. The Peak #1 fraction was purified using two-step column chromatography from a TPBP-acetonitrile solution. The major compound of the fraction was identified as being biphenyl from the 1H NMR and 13C NMR spectra. The ecotoxicity of four degradation products (diphenylborane hydroxide, phenylborane dihydroxide, phenol, and biphenyl) and TPBP towards two marine planktons were assessed. The 48 h LC(50) values of the crustacean, Artemia salina, were 0.13 mg L(-1) for TPBP, 14 mg L(-1) for biphenyl, 17 mg L(-1) for phenol, and > 50 mg L(-1) for the other degradation products. The 72 h EC(50) values of the diatom, Skeletonema costatum, were 0.0022 mg L(-1) for TPBP, 1.2 mg L(-1) for biphenyl, and > 2 mg L(-1) for the other degradation products. Thus, the ecotoxicity of biphenyl and the other degradation products were not high compared to the parent compound, TPBP.

Inputs of chemicals from recreational activities into the Norwegian coastal zone.

Wastewater treatment works effluent is often considered to be one of the most important point sources of a wide range of anthropogenic contaminants to aquatic systems, however, this paper discusses other potential sources. With the aim of establishing the important sources of UV filters, insect repellent and biocides to the aquatic environment, samples were collected from sites with direct (bathing areas and marinas) and indirect (sites receiving wastewater effluent) human influences. Sunscreens containing UV filters are used in large volumes during the summer months and often applied shortly before a person enters the water for swimming activities. The results presented here demonstrate that washing directly from the skin is an important point source of 4 UV filters to the Oslofjord. The insect repellent, diethyl toluamide, was also measured and it was concluded that washing from the skin was not such an important point source into the fjord. Concentrations of the biocide Irgarol 1051 were also measured and were elevated in the small boat marina and surrounding enclosed area. This work demonstrates that man recreational water-based activities are a diffuse source of some contaminants into coastal and fjord environments and this study provides an initial assessment of the levels being released.

Assessment of the risk posed by the antifouling booster biocides Irgarol 1051 and diuron to freshwater macrophytes.

Antifouling paints are used to reduce the attachment of living organisms to the submerged surfaces of ships, boats and aquatic structures, usually by the release of a biocide. Two 'booster' biocides in common use are the triazine herbicide Irgarol 1051 (N-2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine), and diuron (1-(3,4-dichlorophenyl)-3,3-dimethylurea), which are designed to inhibit algal photosynthesis. Previous research has been directed at the effects of these compounds in marine and estuarine environments. In 2001 we sampled the main rivers and shallow freshwater lakes (Broads) of East Anglia UK for Irgarol 1051, its metabolite GS26575 (2-methylamino-4-tert-butylamino-6-amino-s-triazine) and diuron in order to establish the baseline environmental concentrations of these compounds in freshwater systems of eastern UK and to investigate their possible effects on aquatic plants. Irgarol 1051, GS26575 and diuron were found in water samples collected from 21 locations. The highest concentrations were found in the Norfolk and Suffolk Broads in May. The rivers Great Ouse, Wissey, Bure and Yare also contained all three compounds, as did the Great Ouse Cut-off Channel. The toxicity of these biocides to three macrophyte species (Apium nodiflorum, Chara vulgaris, and Myriophyllum spicatum) was investigated. Deleterious effects on relative growth rate, the maximum quantum efficiency (Fv/Fm) of photosystem II and, for Apium, root mass production were found. C. vulgaris was generally most sensitive; growth, especially of roots, was strongly affected in A. nodiflorum; growth rate of M. spicatum was sensitive to diuron. No observed effect concentrations (NOEC) were interpolated using standard toxicological analysis. These were compared with measured environmental concentrations (MEC) to determine the ranges of risk quotients (MEC/NOEC). Both Irgarol 1051 and diuron represented significant risks to A. nodiflorum and C. vulgaris in this area.

Investigating the environmental transport of human pharmaceuticals to streams in the United Kingdom.

The occurrence of 12 selected pharmaceutical compounds and pharmaceutical compound metabolites in sewage treatment works (STW) effluents and surface waters was investigated. The substances selected for the monitoring programme were identified by a risk ranking procedure to identify those substances with the greatest potential to pose a risk to the aquatic environment. STW final effluent and surface water samples were collected from Corby, Great Billing, East Hyde, Harpenden and Ryemeads STWs. Ten of the 12 pharmaceutical compounds were detected in the STW effluent samples: propranolol (100%, median = 76 ng/l), diclofenac (86%, median = 424 ng/l), ibuprofen (84%, median = 3086 ng/l), mefenamic acid (81%, median = 133 ng/l), dextropropoxyphene (74%, median = 195 ng/l), trimethoprim (65%, 70 ng/l), erythromycin (44%, < 10 ng/l), acetyl-sulfamethoxazole (33%, median =< 50 ng/l), sulfamethoxazole (9%, median =< 50 ng/l), tamoxifen (4%, median =< 10 ng/l). In the corresponding receiving streams, fewer compounds and lower concentrations were found: propranolol (87%, median = 29 ng/l), ibuprofen (69%, median = 826 ng/l), mefenamic acid (60%, median = 62 ng/l), dextropropoxyphene (53%, median = 58 ng/l), diclofenac (47%, median =< 20 ng/l), erythromycin (38%, median =< 10 ng/l), trimethoprim (38%, median =< 10 ng/l), acetyl sulfamethoxazole (38%, median =< 50 ng/l). Four human pharmaceutical compounds were detected in samples upstream of the STWs sampled: ibuprofen (57%, median = 181 ng/l), trimethoprim (36%, median < 10 ng/l), erythromycin (17%, median =< 10 ng/l), propranolol (14%, median =< 10 ng/l), suggesting that longer range stream transport of some compounds is possible. The particular STW that was sampled and the month that it was sampled significantly influenced the measured concentrations of several, but not all, substances. There was no significant relationship between usage data and the overall frequency with which different substances were detected. There was however, some evidence to suggest that usage data are positively associated with concentrations of pharmaceuticals in effluent and, particularly, with concentrations measured in surface waters below STWs. These results suggest that most sewage treatment works in England and Wales are likely to be routinely discharging small quantities of pharmaceuticals into UK rivers. None of the pharmaceuticals were found at concentrations that were high enough to cause acute toxic impacts to aquatic organisms. However, insufficient data were available to be able to comment on whether the concentrations measured have the potential to result in more subtle long-term effects on aquatic organisms (e.g. effects on growth, ability to reproduce).

Characterization of estrogenic compounds in water samples collected from United Kingdom estuaries.

This report describes the identification of important estrogenic compounds in surface and sediment pore-water samples collected from the Tyne and Tees estuaries (United Kingdom) through the application of toxicity identification evaluation (TIE) procedures. The Tyne and Tees estuaries represent estuaries that have been historically impacted by industrial activities and continue to receive treated domestic sewage and industrial effluent. In 1998, Dabholm Gut on the Tees received a mixture of treated and untreated effluent, while Howdon sewage treatment works (STW) discharged primary treated effluents. An estrogenically active water sample collected from Howdon STW on the Tyne was shown to contain 17beta-estradiol, androsterone, and an unknown estrogenic compound(s). Most of the activity contained in a sample collected from the Dabholm Gut combined discharges on the Tees was also due to 17beta-estradiol with additional activity from nonylphenol and (tentatively) bis(2-ethylhexyl)phthalate. The only sediment pore-water sample to demonstrate estrogenic activity was collected from Dabholm Gut.

Antifouling paint booster biocides in the UK coastal environment and potential risks of biological effects.

In the yachting sector of the UK antifouling market, organic biocides are commonly added to antifouling preparations to boost performance. Few data presently exist for concentrations of these compounds in UK waters. In this study the concentrations of tributyltin (TBT) and eight booster biocides were measured before and during the 1998 yachting season. The Crouch Estuary, Essex, Sutton Harbour, Plymouth and Southampton Water were chosen as representative study sites for comparison with previous surveys of TBT concentrations. Diuron and Irgarol 1051 were the only organic booster biocides found at concentrations above the limits of detection. Diuron was measured at the highest concentrations, whilst detectable concentrations of both Irgarol 1051 and diuron were determined in areas of high yachting activity (e.g. mooring areas and marinas). Maximum measured values were 1,421 and 6,740 ng/l, respectively. Lower concentrations of both compounds were found in open estuarine areas, although non-antifouling contributions of diuron may contribute to the overall inputs to estuarine systems. TBT was found to be below or near the environmental quality standard (EQS) of 2 ng/l for all samples collected from estuarine areas frequented by pleasure craft alone, but with much higher concentrations measured in some marinas, harbours and in areas frequented by large commercial vessels. Using the limited published environmental fate and toxicity data available for antifouling booster biocides, a comparative assessment to evaluate the risk posed by these compounds to the aquatic environment is described. TBT still exceeds risk quotients by the greatest margins, but widespread effects due to Irgarol 1051 and less so diuron cannot be ruled out (particularly if use patterns change) and more information is required to provide a robust risk assessment.

Toxicity characterisation of organic contaminants in stormwaters from an agricultural headwater stream in south east England.

The transient movement of pesticides at biologically active concentrations during storm events is considered to be a cause of biological impoverishment in some headwater streams. The programme of work described sought to identify compounds that are the cause of toxic effects during such events. Along with targeted pesticide analysis, toxicity identification evaluation (TIE) procedures were used to identify compounds with a demonstrated toxic effect. These procedures were specifically directed towards isolating and attributing toxicity to classes of organic contaminants in samples collected from an English headwater stream during a storm event. The organic load was isolated by means of solid-phase extraction (SPE). Bioassay of the SPE extract at x 100 whole water concentrations confirmed that the samples contained substances toxic to Daphnia magna, although the raw samples were not toxic. Targeted pesticide analysis identified simazine and diuron as the major pesticides present and, using a toxicity unit (TU) approach. were shown to be responsible for a significant amount of the observed concentrate toxicity during a runoff event. However, they were not present in sufficient quantities to be totally responsible for a more toxic later event. By simplification of the SPE isolate using reverse-phase HPLC, fractions from which were tested for toxicity, the cause of concentrate toxicity in the later event was isolated to two discrete fractions. GC-MS analysis of these fractions identified nonylphenol. endosulfan sulphate and pendimethalin as present, with the majority of toxicity attributed to nonylphenol (NP). The main advantage of the TIE approach is that it allows biological active compounds with a demonstrated effect to be identified that may not be selected by more traditional techniques.

Determination of the antifouling agent zinc pyrithione in water samples by copper chelate formation and high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry.

Zinc pyrithione has recently been incorporated into antifouling paints as a booster biocide, which is slowly released into the water as the paint ages. In order to determine concentrations of zinc pyrithione (ZPT) in aqueous samples, a liquid chromatographic method has been developed. Since ZPT interacts with certain reversed-phase packing materials or stainless steel components of the HPLC system, the method uses transchelation of the ZPT into the stable copper(II) complex before analysis by liquid chromatography coupled to atmospheric pressure chemical ionisation mass spectrometry. ZPT was extracted as copper pyrithione using dichloromethane with adequate recovery (77% +/- 17%, n = 6) from 2-l water samples. The limit of detection was calculated to be 20 ng/l, using selected ion monitoring. The analysis of samples collected from various UK marinas showed no detectable concentrations to be present, whilst a laboratory-based study confirmed that this is probably due to the rapid photodegradation of ZPT in seawater.

B-cell and T-cell lymphomas and other associated diseases induced by an infectious DNA viroid-like agent in hamsters (Mesocricetus auratus).

Five epidemics of diffuse, poorly differentiated lymphocytic, immunoblastic, and plasmacytoid lymphoma induced by an infectious, horizontally transmitting viroidlike agent have occurred in two hamster facilities. Incidence summaries and pathologic characteristics of the lymphomas induced in LSH and LVG hamsters are presented. An elevated leukocyte count with a marked increase in neutrophils and a significant decrease in small mononuclear lymphocytes was detected in 5-week-old but not in 10- or 25-week-old LVG hamsters born in the facility contaminated with the lymphoma-inducing agent. Three-week-old LVG hamsters exposed to the contaminated facility showed no similar hematologic change at 5 weeks of age or 5 weeks of exposure. Several associated syndromes, including an intussusception disease, pyelonephritis, inflammatory bowel disease, and body warts associated with the presence of the causative viroidlike agent in the contaminated colonies are described. Details of the epidemiology of the disease, karyology, viral studies, and correlation with several epidemics in other laboratories are presented.

Influence of catecholamines on dexamphetamine-induced changes in locomotor activity.

In the mouse, central noradrenaline receptor stimulation by clonidine, or intracerebroventricular injection of noradrenaline or alpha-methylnoradrenaline, caused marked enhancement of the locomotor stimulant effects of dexamphetamine in doses that were without effect when given alone. A minimally locomotor-stimulant dose of apomorphine reduced the effect of dexamphetamine. Pimozide and phenoxybenzamine each virtually abolished locomotor stimulation after dexamphetamine, while FLA63 caused significant reduction. Phenoxybenzamine also abolished the enhancement by clonidine. The intensity of the dexamphetamine effect was dose-related, while in the case of apomorphine the duration rather than the intensity was related to the dose administered. Clonidine potentiated apomorphine locomotor stimulation; following this drug combination, the nature of the movements more closely resembled those seen after dexamphetamine. The results suggest the involvement of both noradrenaline and dopamine in the dexamphetamine response.