The U.S. federal framework for research on endocrine disruptors and an analysis of research programs supported during fiscal year 1996.

The potential health and ecological effects of endocrine disrupting chemicals has become a high visibility environmental issue. The 1990s have witnessed a growing concern, both on the part of the scientific community and the public, that environmental chemicals may be causing widespread effects in humans and in a variety of fish and wildlife species. This growing concern led the Committee on the Environment and Natural Resources (CENR) of the National Science and Technology Council to identify the endocrine disruptor issue as a major research initiative in early 1995 and subsequently establish an ad hoc Working Group on Endocrine Disruptors. The objectives of the working group are to 1) develop a planning framework for federal research related to human and ecological health effects of endocrine disrupting chemicals; 2) conduct an inventory of ongoing federal research programs; and 3) identify research gaps and develop a coordinated interagency plan to address priority research needs. This communication summarizes the activities of the federal government in defining a common framework for planning an endocrine disruptor research program and in assessing the status of the current effort. After developing the research framework and compiling an inventory of active research projects supported by the federal government in fiscal year 1996, the CENR working group evaluated the current federal effort by comparing the ongoing activities with the research needs identified in the framework. The analysis showed that the federal government supports considerable research on human health effects, ecological effects, and exposure assessment, with a predominance of activity occurring under human health effects. The analysis also indicates that studies on reproductive development and carcinogenesis are more prevalent than studies on neurotoxicity and immunotoxicity, that mammals (mostly laboratory animals) are the main species under study, and that chlorinated dibenzodioxins and polychlorinated biphenyls are the most commonly studied chemical classes. Comparison of the inventory with the research needs should allow identification of underrepresented research areas in need of attention.

Food chain transfer and potential renal toxicity of mercury to small mammals at a contaminated terrestrial field site.

Mercury concentrations were determined in surface soil and biota at a contaminated terrestrial field site and were used to calculate transfer coefficients of mercury through various compartments of the ecosystem based on trophic relationships. Mercury concentrations in all compartments (soil, vegetation, invertebrates, and small mammals) were higher than mercury concentrations in corresponding samples at local reference sites. Nonetheless, mercury concentrations in biota did not exceed concentrations in the contaminated surface soil, which averaged 269 µg g(-1). Plant tissue concentrations of mercury were low (0.01 to 2.0 µg g(-1)) and yielded soil to plant transfer coefficients ranging from 3.7×10(-5) for seeds to 7.0×10(-3) for grass blades. Mercury concentrations in invertebrates ranged from 0.79 for harvestmen (Phalangida) to 15.5 µg g(-1) for undepurated earthworms (Oligochaeta). Mean food chain transfer coefficients for invertebrates were 0.88 for herbivores/omnivores and 2.35 for carnivores. Mean mercury concentrations in target tissue (kidney) were 1.16±1.16 µg g(-1) for the white-footed mouse (Peromyscus leucopus), a granivore, and 38.8±24.6 µg g(-1) for the shorttail shrew (Blarina brevicauda), an insectivore. Transfer coefficients for diet to kidney were 0.75 and 4.40 for P. leucopus and B. brevicauda, respectively. A comparison of kidney mercury residues measured in this study with values from controlled laboratory feeding studies from the literature indicate that B. brevicauda but not P. leucopus may be ingesting mercury at levels that are nephrotoxic.

Small mammals as monitors of environmental contaminants.

The merit of using small mammals as monitors of environmental contaminants was assessed using data from the published literature. Information was located on 35 species of small mammals from 7 families used to monitor heavy metals, radionuclides, and organic chemicals at mine sites, industrial areas, hazardous and radioactive waste disposal sites, and agricultural and forested land. To document foodchain transfer of chemicals, concentrations in soil, vegetation, and invertebrates, where available, were included. The most commonly trapped North American species were Peromyscus leucopus, Blarina brevicauda, and Microtus pennsylvanicus. In these species, exposure to chemicals was determined from tissue residue analyses, biochemical assays, and cytogenetic assays. Where enough information was available, suitable target tissues, or biological assays for specific chemicals were noted. In general, there was a relationship between concentrations of contaminants in the soil or food, and concentrations in target tissues of several species. This relationship was most obvious for the nonessential heavy metals, cadmium, lead, and mercury and for fluoride. Kidney was the single best tissue for residue analyses of inorganic contaminants. However, bone should be the tissue of choice for both lead and fluorine. Exposure to lead was also successfully documented using biochemical and histopathological endpoints. Bone was the tissue of choice for exposure to 90Sr, whereas muscle was an appropriate tissue for 137Cs. For organic contaminants, exposure endpoints depended on the chemical(s) of concern. Liver and whole-body residue analyses, as well as enzyme changes, organ histology, genotoxicity, and, in one case, population dynamics, were successfully used to document exposure to these contaminants. Based on information in these studies, each species' suitability as a monitor for a specific contaminant or type of contaminant was evaluated and subsequently ranked. A relationship between contaminant exposure and trophic level emerged. Insectivores (shrews) had the highest levels of contaminants, followed by omnivores (cricetid mice) with intermediate levels, and herbivores (voles) with the lowest levels. A substantial number of these biomonitoring studies using small mammals collectively point to the importance of food habits and habitat of small mammals, and their availability and abundance as factors that should influence species selection for monitoring studies. The type of contaminants under consideration as well as the appropriateness of the endpoints selected are important factors to consider when deciding whether or not to include small mammals in biomonitoring studies.

Microbial degradation of trichloroethylene in the rhizosphere: potential application to biological remediation of waste sites.

The possibility that vegetation may be used to actively promote microbial restoration of chemically contaminated soils was tested by using rhizosphere and nonvegetated soils collected from a trichloroethylene (TCE)-contaminated field site. Biomass determinations, disappearance of TCE from the headspace of spiked soil slurries, and mineralization of [14C]TCE to 14CO2 all showed that microbial activity is greater in rhizosphere soils and that TCE degradation occurs faster in the rhizosphere than in the edaphosphere. Thus, vegetation may be an important variable in the biological restoration of surface and near-surface soils.

Benzoquinolinediones: activity as insect teratogens.

Morphological abnormalities including extra compound eyes, extra heads, and distally duplicated legs were generated in cricket embryos by treating eggs with single doses of either benz[g]isoquinoline-5,10-dione or benzo[h]quinoline-5,6-dione. Slight structural modifications of the molecules resulted in a loss of teratogenic activity, although embryotoxicity occurred. These potent insect teratogens can be used for analysis of developmental events during embryogenesis.

Use of the cricket embryo (Acheta domesticus) as an invertebrate teratology model.

Embryos of the cricket Acheta domesticus (L.) have been shown by bioassay to develop gross morphological abnormalities after exposure to a number of complex organic mixtures as well as to display a critical period of teratogen sensitivity and an ability to metabolize xenobiotics during development. Because the assay is simple, inexpensive, short-term (less than two weeks), and objective, it could be useful as an in vivo screen in a hierarchical approach to teratogen detection. Further investigation of cricket embryo responses to known teratogens is needed to establish the predictive value of this assay.

Pharmacodynamic behavior of [14C]acridine in the cricket Acheta domesticus (L.).

Cuticular and gastrointestinal penetration, in vivo metabolism, and excretion of [14C]acridine were investigated in the nymphal cricket Acheta domesticus (L.) to find a pharmacodynamic basis for this insect's differential susceptibility to acridine at different life stages. Topically applied [14C]acridine readily penetrated the cuticular exoskeleton of nymphs (half-time of penetration, 48 min). Radiolabeled compounds appeared in the hemolymph within 0.5 h after ingestion of [14C]acridine and continued to move across the gut wall for 7.5 h. The biological half-time was 18 h and the rate constant for elimination was 0.039 h-1 after ingestion. Within 5 d after dosing, 97% of the dose was excreted. Several metabolites were present in the feces of nymphs fed [14C]acridine, and less than 13% of the extractable radioactivity was parent compound. The cuticle and the gastrointestinal tract proved to be ineffective barriers to acridine entry in A, domesticus. However, the ability to readily metabolize and excrete acridine probably contributes to the higher acridine tolerance observed in the nymphs and adults than in the eggs, which are susceptible to toxic effects.

Chemical impurity produces extra compound eyes and heads in crickets.

A chemical impurity isolated from commercially purchased acridine causes cricket embryos to develop extra compound eyes, branched antennae, extra antennae, and extra heads. Purified acridine does not produce similar duplications of cricket heads or head structures nor do the substituted acridines proflavine, acriflavine, or acridine orange. A dose-response relation exists such that the number and severity of abnormalities increase with increasing concentration of the teratogen.