Joanne L. Parrott

Pharmaceuticals and Personal Care Products in the Environment: What Are the Big Questions?

Background: Over the past 10–15 years, a substantial amount of work has been done by the scientific, regulatory, and business communities to elucidate the effects and risks of pharmaceuticals and personal care products (PPCPs) in the environment. Objective: This review was undertaken to identify key outstanding issues regarding the effects of PPCPs on human and ecological health in order to ensure that future resources will be focused on the most important areas. Data sources: To better understand and manage the risks of PPCPs in the environment, we used the “key question” approach to identify the principle issues that need to be addressed. Initially, questions were solicited from academic, government, and business communities around the world. A list of 101 questions was then discussed at an international expert workshop, and a top-20 list was developed. Following the workshop, workshop attendees ranked the 20 questions by importance. Data synthesis: The top 20 priority questions fell into seven categories: a) prioritization of substances for assessment, b) pathways of exposure, c) bioavailability and uptake, d) effects characterization, e) risk and relative risk, f ) antibiotic resistance, and g) risk management. Conclusions: A large body of information is now available on PPCPs in the environment. This exercise prioritized the most critical questions to aid in development of future research programs on the topic.

Toxicity of oil sands to early life stages of fathead minnows (Pimephales promelas)

The present study examines the effects of exposure to oil sands on the early life stages (ELS) of fathead minnows (Pimephales promelas). Sediments within and outside natural oil sand deposits were collected from sites along the Athabasca River (AB, Canada). The ELS toxicity tests were conducted with control water, natural oil sands, reference sediments, and oil-refining wastewater pond sediments. Eggs and larvae were exposed to 0.05 to 25.0 g sediment/L and observed for mortality, hatching, malformations, growth, and cytochrome P4501A induction as measured by immunohistochemistry. Natural bitumen and wastewater pond sediments caused significant hatching alterations and exposure-related increases in ELS mortality, malformations, and reduced size. Larval deformities included edemas, hemorrhages, and spinal malformations. Exposure to reference sediments and controls showed negligible embryo mortality and malformations and excellent larval survival. Sediment analyses using gas chromatography-mass spectrometry revealed high concentrations of alkyl-substituted polyaromatic hydrocarbons (PAHs) compared to unsubstituted PAHs in natural oil sands (220-360 microg/g) and oil-mining wastewater pond sediments (1,300 microg/g). The ELS sediment toxicity tests are rapid and sensitive bioassays that are useful in the assessment of petroleum toxicity to aquatic organisms.

Altered reproduction in fish exposed to pulp and paper mill effluents: roles of individual compounds and mill operating conditions.

For the last 20 years, studies conducted in North America, Scandinavia, and New Zealand have shown that pulp and paper mill effluents affect fish reproduction. Despite the level of effort applied, few leads are available regarding the factors responsible. Effluents affect reproduction in multiple fish species, as evidenced by decreased gonad size, decreased circulating and gonadal production of reproductive steroids, altered expression of secondary sex characteristics, and decreased egg production. Several studies also have shown that effluent constituents are capable of accumulating in fish and binding to sex steroid receptors/ binding proteins. Studies aimed at isolating biologically active substances within the pulping and papermaking process have provided clues about their source, and work has progressed in identifying opportunities for in-mill treatment technologies. Following comparisons of manufacturing processes and fish responses before and after process changes, it can be concluded that effluent from all types of mill processes are capable of affecting fish reproduction and that any improvements could not be attributed to a specific process modification (because mills normally performed multiple modifications simultaneously). Improved reproductive performance in fish generally was associated with reduced use of molecular chlorine, improved condensate handling, and liquor spill control. Effluent biotreatment has been effective in reducing some effects, but biotreated effluents also have shown no difference or an exacerbation of effects. The role of biotreatment in relation to effects on fish reproduction remains unclear and needs to be resolved.

Profiling oil sands mixtures from industrial developments and natural groundwaters for source identification.

The objective of this study was to identify chemical components that could distinguish chemical mixtures in oil sands process-affected water (OSPW) that had potentially migrated to groundwater in the oil sands development area of northern Alberta, Canada. In the first part of the study, OSPW samples from two different tailings ponds and a broad range of natural groundwater samples were assessed with historically employed techniques as Level-1 analyses, including geochemistry, total concentrations of naphthenic acids (NAs) and synchronous fluorescence spectroscopy (SFS). While these analyses did not allow for reliable source differentiation, they did identify samples containing significant concentrations of oil sands acid-extractable organics (AEOs). In applying Level-2 profiling analyses using electrospray ionization high resolution mass spectrometry (ESI-HRMS) and comprehensive multidimensional gas chromatography time-of-flight mass spectrometry (GC × GC-TOF/MS) to samples containing appreciable AEO concentrations, differentiation of natural from OSPW sources was apparent through measurements of O2:O4 ion class ratios (ESI-HRMS) and diagnostic ions for two families of suspected monoaromatic acids (GC × GC-TOF/MS). The resemblance between the AEO profiles from OSPW and from 6 groundwater samples adjacent to two tailings ponds implies a common source, supporting the use of these complimentary analyses for source identification. These samples included two of upward flowing groundwater collected <1 m beneath the Athabasca River, suggesting OSPW-affected groundwater is reaching the river system.

A Decade of Research on the Environmental Impacts of Pulp and Paper Mill Effluents in Canada: Sources and Characteristics of Bioactive Substances

This article is a review of research efforts over the last decade on the sources and characteristics of substances in Canadian pulp mill effluents associated with two responses in fish: (1) induction of detoxification enzymes and (2) reproductive effects. The initial uncertainty regarding the role of chlorine bleaching and dioxins in these responses was resolved by the mid 1990s, when it was determined that effects were not correlated with effluent adsorbable organic halogen (AOX) levels and that releases of dioxins had decreased substantially. In the mid 1990s researchers were able to partially attribute enzyme activity induction in fish to wood components, while other studies showed individual wood extractives had the potential to affect fish reproduction. A lack of correlation between threshold reproductive responses and effluent concentrations indicated additional unidentified compounds and mechanisms were involved. In the late 1990s, source identification approaches in concert with the development of mechanistically linked in vitro and in vivo bioassays showed multiple compounds are affecting production and signaling of sex steroids in fish. These substances are bioavailable and accumulated rapidly, consistent with the body of evidence that has shown a sustained exposure is required to produce both elevated enzyme activity and depressions in sex steroid levels. The patterns of these substances in effluents and fish tissues are not correlated with production type or effluent treatment. Collectively, these findings show that bioactive substances originate from wood and are derived from lignin and/or terpenoids, they are liberated during pulp digestion, and in kraft mills they are present in black liquor and chemical recovery condensates. Additional bioactive substances are also present in bleachery effluents containing residual lignin. The lack of a definitive identification of the responsible compounds has prevented an evaluation of the effectiveness of industry-wide process changes. Continued research into the identities, origins, and environmental fate of these substances and the efficacy of effluent treatment is required to determine their significance and relationship to the existing impacts of effluents from pulp and paper mills in Canadian aquatic ecosystems. Completion of this review would not have been possible without the helpful comments and scientific reviews provided by Drs. John Carey, Jim Maguire, Kelly Munkittrick, Glen Van Der Kraak, and Olöf Sandstrom.

CYP1A Induction and Blue Sac Disease in Early Life Stages of White Suckers (Catostomus commersoni) Exposed to Oil Sands

The objectives of this study were to evaluate the influence of natural oil sands on the early developmental stages of white sucker (Catostomus commersoni) and to determine whether biochemical responses in this species were similar to native fish caught in the Athabasca Oil Sands area. Early life stage (ELS) sediment toxicity tests were conducted using controls, reference sediments, natural oil sands, and industrially contaminated (wastewater pond) sediments collected from sites along the Athabasca River, Alberta (Canada). Eggs and larvae were observed for mortality, hatching, deformities, growth, and cytochrome P-4501A (CYP1A) activity using immunohistochemistry. E-Nat-, S-Nat-, and wastewater pond sediment-exposed groups showed significant premature hatching, reduced growth, and exposure-dependent increases in ELS mortality and larval malformations relative to controls. The most common larval deformities included edemas (pericardial, yolk sac, and subepidermal), hemorrhages, and spinal defects. Juveniles exposed to oil sands and wastewater pond sediments (96 h) demonstrated significantly increased 7-ethoxyresorufin–O-deethylase (EROD) activity (30- to 50-fold) as compared to controls. Reference sediment-exposed groups and water controls demonstrated reliable embryo and larval survival, minimal malformations, and negligible CYP1A staining. These observed signs of blue sac disease (ELS mortality, malformations, growth reductions, CYP1A activity induction) may produce deleterious reproductive effects in natural fish populations exposed to oil sands mixtures. This research was funded by grants from the Toxic Substances Research Initiative (Project 187) through Health Canada, Panel on Energy Research and Development, and the Natural Sciences and Engineering Council of Canada (PVH). M. Colavecchia was supported by scholarships awarded by OGS, Canadian Network of Toxicology Centres, and Petro-Canada. The authors thank the following individuals for their technical assistance with the field sampling program: R. Neurtherander (NWRI), B. Crosley, M.Conly (CWS), Golder Associates (K. Allen and M. Ezekiel), Regional Aquatics Monitoring Program, Suncor Energy (A. Cummins), Syncrude Canada (T. VanMeer, N. Rutley), M. Bowerman, and A. Winchester (Queens University). We thank G. Fodor (DFO), Dr. P. Akhtar (Queens University), B. Blunt, M. Baker (NWRI), S. Cagampan, and S. Backus (NLET) for their enthusiastic laboratory assistance. Portions of this research were presented at the 2003 Annual Aquatic Toxicity Workshop (Thirtieth Annual Meeting Abstracts, p. 90). Useful comments by Dr. S. Kacew and two anonymous reviewers helped improve an earlier draft of this article.

Environmental risk assessment for the serotonin re-uptake inhibitor fluoxetine: Case study using the European Risk Assessment Framework

The serotonin re-uptake inhibitor fluoxetine was selected for an environmental risk assessment, using the most recent European guideline (EMEA 2006) within the European Union (EU)-funded Environmental Risk Assessment of Pharmaceuticals (ERAPharm) project due to its environmental persistence, acute toxicity to nontarget organisms, and unique pharmacokinetics associated with a readily ionizable compound. As a widely prescribed psychotropic drug, fluoxetine is frequently detected in surface waters adjacent to urban areas because municipal wastewater effluents are the primary route of entry to aquatic environments. In Phase I of the assessment, the initial predicted environmental concentration of fluoxetine in surface water (initial PEC(SW)) reached or exceeded the action limit of 10 ng/L, when using both a default market penetration factor and prescription data for Sweden, Germany, and the United Kingdom. Consequently, a Phase II risk assessment was conducted in which green algae were identified as the most sensitive species with a NOEC of <0.6 microg/L. From this value, a predicted no effect concentration for surface waters (PNEC(SW)) of 0.012 microg/L was derived. The PEC/PNEC ratio was above the trigger value of 1 in worst-case exposure scenarios indicating a potential risk to the aquatic compartment. Similarly, risks of fluoxetine for sediment-dwelling organisms could not be excluded. No risk assessment was conducted for the terrestrial compartment due to a lack of data on effects of fluoxetine on soil organisms. The need for a separate risk assessment for the main metabolite of fluoxetine, norfluoxetine, was not conducted because of a lack of fate and effect studies. Based on published data, fluoxetine and norfluoxetine appeared to have a low to moderate bioaccumulation potential, which should be confirmed in formal studies according to OECD guidelines. Exposure assessments for fluoxetine according to the current framework rely heavily on K(OC) and K(OW) values. This approach is problematic, because fluoxetine is predominantly a cationic substance at environmental pH values. Consequently, the fate of fluoxetine (and other ionic substances) cannot be predicted using partition coefficients established for nonionic compounds. Further, published estimates for partition coefficients of fluoxetine vary, resulting in considerable uncertainties in both the exposure and environmental risk assessments of fluoxetine.

Using reproductive endpoints in small forage fish species to evaluate the effects of athabasca oil sands activities

The main objective of this study was to evaluate the influence of naturally occurring oil sands-related compounds (OSRC) on reproductive function in fish in order to assess the impacts of anthropogenic point-source inputs. The health of slimy sculpin (Cottus cognatus) and pearl dace (Semotilus margarita) collected from the Alberta Athabasca Oil Sands (Canada) watershed were examined. Two rivers were selected for study: the Steepbank and the Ells. These rivers originate outside the oil sands formation, where fish are unexposed (Ref), exposed to naturally occurring oil sands-related compounds (Nat), or exposed to naturally occurring compounds as well as adjacent to surface mining activity (Dev). Assessment endpoints included gonadosomatic indices (GSI), fecundity, and in vitro gonadal steroid production. In vitro gonadal incubations demonstrated lower levels of steroid production at sites along the Steepbank River within the oil sands deposit. Hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity, an indicator of exposure to OSRC, was elevated twofold at the site with natural compounds and up to 10-fold at the site adjacent to development compared to EROD activity in fish from the reference site. Fish collected in the Ells River had a threefold induction in EROD activity but no significant reduction in steroid production when compared to reference fish. No consistent alterations in gonadal development were seen in fish collected from sites within the oil sands deposit. This research in the Athabasca River basin provides baseline information of the health of fish populations within the oil sands deposit prior to further development in the area.

A Decade of Research on the Environmental Impacts of Pulp and Paper Mill Effluents in Canada: Field Studies and Mechanistic Research

Studies conducted in Sweden in the early 1980s provided some of the first evidence that effluents from some pulp mills were capable of inducing toxic responses in fish at very low concentrations in the receiving environment. In response to these findings, studies were initated in Canada and impacts of primary treated bleached kraft mill effluent on reproductive function in fish were found. Reproductive impacts in fish were not limited to mills that used chlorine in the bleaching process and were also evident at some mills that employed secondary effluent treatment. In 1992, new federal regulations were passed under the Canadian Environmental Protection Act to control releases of dioxins and furans, and a new Pulp and Paper Effluent Regulation under the Fisheries Act set stricter limits for biological oxygen demand and total suspended solids. Very importantly, the new regulations included requirements for environmental effects monitoring (EEM) at all mill sites. This allowed the effectiveness of the control limits in protecting fish, fish habitat, and human use of fisheries resources to be assessed. At the same time, the Minister of the Environment launched an intensive government, industry, and university research program. Results from this research program along with feedback from the EEM program would then be used to define what additional control actions might be necessary. This article reviews the field studies and mechanistic research conducted in Canada following the implementation of the new federal regulations. Great progress has been made in this area, first demonstrating reproductive effects at various locations, then determining the mechanisms responsible for the reproductive effects at specific sites, followed by the demonstration of partial recovery in reproductive function following process and treatment changes in response to the new regulations. However, it is clear from the results of the first two cycles of the EEM program that mill effluents still affect the local receiving environments at a number of locations across Canada, and that continued research combining field studies, bioassay application, and chemical identification is required. Completion of this assessment would not have been possible without the helpful comments and scientific reviews provided by Drs. John Carey, Jim Maguire, Kelly Munkittrick, Glen Van Der Kraak, Jim Sherry, and Olöf Sandstrom.

Life-Cycle Exposure of Fathead Minnows to a Mixture of Six Common Pharmaceuticals and Triclosan

Fathead minnows were exposed for a life cycle to environmentally relevant concentrations of a mixture of six common pharmaceuticals and one personal care product (nominal concentrations: 1,000, 300, 100, 30, or 10 ng/L). Mean measured concentrations of each chemical in the highest fish exposure aquaria were: naproxen 793 ng/L, gemfibrozil 662 ng/L, diclofenac 331 ng/L, ibuprophen 217 ng/L, triclosan 115 ng/L, salicylic acid 67 ng/L, and acetaminophen (chemical analysis inconclusive, nominal 1000 ng/L). Fish exposed for a life cycle even to the highest concentrations of the six pharmaceuticals and personal care product (PPCP) mixture showed no significant changes in growth and development compared to control. Length, weights, condition factors, liver weights, and gonad weights of PPCP-exposed fish were similar to water and solvent controls (0.000005% ethanol v/v). There were no marked effects of PPCP mixture exposure on external sex characteristics of the fish or on egg production. The only parameter that appeared to be affected was percent larval deformities in F1, which showed a significant increase in the 100- and 300-ng/L (nominal) PPCP mixture. Larvae from control fish had 4.7% (water controls) and 3.4% (solvent controls) deformities, compared to 9.3% in the 100-ng/L (nominal) PPCP mixture and 9.2% deformities in the 300-ng/L (nominal) PPCP mixture. Chronic exposure to environmentally relevant concentrations of seven PPCP most often detected in Canadian municipal wastewater effluents (MWWE) did not appear to affect fathead minnow survival, growth, or egg production, although it produced quantitative increases in deformities in the F1 generation.