Modulation of steroidogenesis by coastal waters and sewage effluents of Hong Kong, China, using the H295R assay

BACKGROUND, AIM, AND SCOPE: The presence of a variety of pollutants in the aquatic environment that can potentially interfere with the production of sex steroid hormones in wildlife and humans has been of increasing concern. The aim of the present study was to investigate the effects of extracts from Hong Kong marine waters, and influents and effluents from wastewater treatment plants on steroidogenesis using the H295R cell bioassay. After exposing H295R cells to extracts of water, the expression of four steroidogenic genes and the production of three steroid hormones were measured. MATERIALS AND METHODS: Water samples were collected during the summer of 2005 from 24 coastal marine areas and from the influents and effluents of two major waste water treatment plants (WWTPs) in Hong Kong, China. Samples were extracted by solid phase extraction (SPE). H295R cells were exposed for 48 h to dilutions of these extracts. Modulations of the expression of the steroidogenic genes CYP19, CYP17, 3betaHSD2, and CYP11beta2 were determined by measuring mRNA concentrations by real-time polymerase chain reaction (Q-RT-PCR). Production of the hormones progesterone (P), estradiol (E2), and testosterone (T) was quantified using enzyme linked immunosorbent assays (ELISA). RESULTS: Extracts from samples collected in two fish culture areas inhibited growth and proliferation of H295R cells at concentrations greater or equal to 10(5) L equivalents. The cells were exposed to the equivalent concentration of active substances in 10,000 L of water. Thus, to observe the same level of effect as observed in vitro on aquatic organisms would require a bioaccumulation factor of this same magnitude. None of the other 22 marine samples affected growth of the cells at any dilution tested. Twelve of the marine water samples completely inhibited the expression of CYP19 without affecting E2 production; inhibition of CYP17 expression was observed only in one of the samples while expression of CYP11beta2 was induced as much as five- and ninefold after exposure of cells to extracts from two locations. The expression of the progesterone gene 3betaHSD2 was not affected by any of the samples; only one sample induced approximately fourfold the production of E2. Although more than twofold inductions were observed for P and T production, none of these values were statistically significant to conclude effects on the production of these two hormones. While influents from WWTPs did not affect gene expression, an approximately 30% inhibition in the production of E2 and a 40% increase in P occurred for the exposure with influents from the Sha Tin and Stonecutters WWTPs, respectively. Effluents from WWTPs did not affect the production of any of the studied hormones, but a decrement in the expression of the aldosterone gene CYP11beta2 was observed for the Sha Tin WWTP exposure. No direct correlation could be established between gene expression and hormone production. DISCUSSION: Observed cytotoxicity in the two samples from fish culture areas suggest the presence of toxic compounds; chemical analysis is required for their full identification. Although effluents from WWTPs did not affect hormone production, other types of endocrine activity such as receptor-mediated effects cannot be ruled out. Interactions due to the complexity of the samples and alternative steroidogenic pathways might explain the lack of correlation between gene expression and hormone production results. CONCLUSIONS: Changes observed in gene expression and hormone production suggest the presence in Hong Kong coastal waters of pollutants with endocrine disruption potential and others of significant toxic effects. The aromatase and aldosterone genes seem to be the most affected by the exposures, while E2 and P are the hormones with more significant changes observed. Results also suggest effectiveness in the removing of compounds with endocrine activity by the WWTPs studied, as effluent samples did not significantly affect hormone production. The H295R cell showed to be a valuable toll in the battery required for the analysis of endocrine disrupting activities of complex environmental samples. RECOMMENDATIONS AND PERSPECTIVES: Due to the intrinsic complexity of environmental samples, a combination of analytical tools is required to realistically assess environmental conditions, especially in aquatic systems. In the evaluation of endocrine disrupting activities, the H295R cell bioassay should be used in combination with other genomic, biological, chemical, and hydrological tests to establish viable modes for endocrine disruption and identify compounds responsible for the observed effects.     

Evidence for the "grasshopper" effect and fractionation during long-range atmospheric transport of organic contaminants

Although there is indisputable evidence that long-range atmospheric transport (LRAT) of organic contaminants occurs on a global scale, uncertainties remain about the detailed mechanism and extent of this phenomenon as well as the physical-chemical properties which facilitate LRAT. In this study, we discuss how mass balance models and monitoring data can contribute to a fuller understanding of the mechanism and extent of LRAT. Specifically we address the issues of "grasshopping" or "hopping" (the extent to which molecules are subject to multiple hops as distinct from a single emission-deposition event) and "global fractionation" (the differing behavior of chemicals as they are transported). It is shown that simple mass balance models can be used to assist the interpretation of monitoring data while also providing an instrument that can be used to assess the LRAT potential and the extent of hopping that organic substances may experience. The available evidence supports the notion that many persistent organic pollutants experience varying degrees of "hopping" during their environmental journey and as a consequence become fractionated with distance from source.     

Trace metals in the hair of habitants of the Ok Tedi region, Papua New Guinea

It has long been known that mining activity can markedly change the level and distribution of certain heavy metals in the adjacent environment. This pollution can be quite widespread and long lasting and often has deleterious effects on the health of local populations. In the present study scalp hair was used as the biopsy material because of its ease of collection and long history of use in this connection. Hair was collected from all the local villages in the vicinity of the mine site, as well as from Papua New Guinean nationals from other provinces, and European expatriates who were employed by the mining company and who were resident in the area. Hair from local people showed a remarkably high iron content by comparison with previously studied populations. The extreme variations in hair iron levels were reflected in the differential distribution of levels according to location, age and sex. Hair cadmium was also high in the population studied. Hair copper, lead, zinc and mercury all appeared to be within 'normal' limits by comparison with other general populations. These results are discussed in the context of the environmental and social impact of the mining operations on the local people.     

Chemical fractionation of copper, lead and zinc in ombrotrophic peat

Measurement of operationally defined chemical fractions in ombrotrophic peat samples provides information not obtained by total metal extractions. Examination of such sequential data permits interpretation of process dynamics. Results for copper, lead and zinc chemical fractionation of peat profiles from Ringinglow Bog, near Sheffield, Great Britain, are reported and discussed. Lead and zinc share similar patterns of partitioning. A considerable proportion of these records is in a form predisposed to transformation and mobilisation. In contrast, much of the copper is comparatively immobile, it being associated with the more chemically intractable fractions. The results suggest that peat copper deposition records may be used for reconstruction of pollution history. The use of lead and zinc records for these purposes is problematic.     

Bioavailability of persistent organic pollutants in soils and sediments--a perspective on mechanisms, consequences and assessment

It has been observed that as soil-pollutant contact time increases, pollutant bioavailability and extractability decreases. This phenomenon has been termed 'ageing'. Decreased chemical extractability with increased soil-chemical contact time is evident where both 'harsh' techniques, e.g. dichloromethane Soxhlet extraction, and 'non-exhaustive' techniques, e.g. butanol shake extraction, have been used. It has also been observed that the amount of chemical extracted by these techniques varies considerably over time. Similarly, decreases in bioavailability with increased soil-pollutant contact time have been described in bacterial, earthworm and other organism studies. From these investigations, it has been shown that the fraction of pollutant determined to be bioavailable can vary between organisms. Thus, there is an immediate definition problem, what is bioavailability? Additionally, if bioavailability is to be assessed by a chemical means, which organisms should (or can) be mimicked by the extraction procedure? This review provides a background to the processes inherent to ageing, a discussion of its consequences on bioavailability and ends with some reflections on the appropriateness of chemical extraction techniques to mimic bioavailability     

The global re-cycling of persistent organic pollutants is strongly retarded by soils

'Persistent organic pollutants' (POPs) are semi-volatile, mobile in the environment and bioaccumulate. Their toxicity and propensity for long-range atmospheric transport (LRAT) has led to international bans/restrictions on their use/release. LRAT of POPs may occur by a 'single hop' or repeated temperature-driven air-surface exchange. It has been hypothesised that this will result in global fractionation and distillation-with condensation and accumulation in polar regions. Polychlorinated biphenyls (PCBs)--industrial chemicals banned/restricted in the 1970s--provide a classic illustration of POP behaviour. A latitudinally-segmented global PCB inventory has been produced, which shows that approximately 86% of the 1.3 x 10(6) tonnes produced was used in the temperate industrial zone of the northern hemisphere. A global survey of background surface soils gives evidence for 'fractionation' of PCBs. More significantly, however, very little of the total inventory has 'made the journey' via primary emission and/or air-surface exchange and LRAT out of the heavily populated source regions, in the 70 years since PCBs were first produced. Soils generally occlude PCBs, especially soils with dynamic turnover of C/bioturbation/burial mechanisms. This limits the fraction of PCBs available for repeated air-soil exchange. The forested soils of the northern hemisphere, and other C-rich soils, appear to be playing an important role in 'protecting' the Arctic from the advective supply of POPs. Whilst investigations on POPs in remote environments are important, it is imperative that researchers also seek to better understand their release from sources, persistence in source regions, and the significant loss mechanisms/global sinks of these compounds, if they wish to predict future trends.     

Bound pesticide residues in soils: a review

This article is a review of the current state of knowledge regarding the formation and biological/environmental significance of bound pesticide residues in soils. We begin by defining various terms used in our discussions and identifying the types/classes of pesticides which may be added to soil and interact with it. We then consider various soil properties and aspects of land management which will influence the nature and degree of the soil-pesticide association and discuss the possible physical and chemical binding mechanisms. We then move on to consider the role of microorganisms and other forms of soil biota in bound residue formation and the bioavailability of soil-borne pesticide residues. The review ends with a consideration of the significance of bound pesticide residues.     

Assessing Landscape Condition Relative to Water Resources in the Western United States: A Strategic Approach

The Environmental Monitoring and Assessment Program (EMAP) is proposing an ambitious agenda to assess the status of streams and estuaries in a 12-State area of the western United States by the end of 2003. Additionally, EMAP is proposing to access landscape conditions as they relate to stream and estuary conditions across the west. The goal of this landscape project is to develop a landscape model that can be used to identify the relative risks of streams and estuaries to potential declines due to watershed-scale, landscape conditions across the west. To do so, requires an understanding of quantitative relationships between landscape composition and pattern metrics and parameters of stream and estuary conditions. This paper describes a strategic approach for evaluating the degree to which landscape composition and pattern influence stream and estuary condition, and the development and implementation of a spatially-distributed, landscape analysis approach.     

INTERSTATE AND INTERNATIONAL AQUIFERS1

Many important groundwater aquifers cross state and national boundaries. The flow of water in these aquifers is not influenced by the boundaries but may be materially influenced by mans activities on one or both sides of a boundary. Interstate and international problems may develop because of excessive groundwater lowering on one side of a boundary affecting water users on the opposite side of the line. Similarly, intensive groundwater development along a surface stream may influence the amount of surface water that flows across a boundary. A third type of problem may develop when pumping on one side of the boundary induces poor quality water into an aquifer on the other side of the boundary. Several specific interstate and international aquifer problems are briefly described.     

An ecological economics approach to the management of a multi-purpose coastal wetland

Three interrelated management problems—eutrophication of multiple use shallow lakes, sea level rise and flood risk mitigation and tourism pressures—are analysed in the context of an internationally important wetland area, the Norfolk and Suffolk Broads in the UK. The ecological-economic research findings presented should provide essential information to underpin the regulatory and management process in this internationally important conservation area. The relevant authority somehow has to integrate the maintenance of public navigation rights, nature conservation, and tourism promotion in a highly dynamic ecosystems setting. Because of the stakeholder conflicts, potential and actual, a more inclusionary decision-making procedure is required, and is currently being implemented.