[Ecological risk assessment and its application].

Ecological risk assessment, closely linked to environmental toxicology, is becoming one of the most widely expanding disciplines in environmental science. A number of frameworks for ecological risk assessment have been developed by the USEPA, Environment Canada and European Commission in recent years. This review described the common elements, the process and steps of ecological risk assessment. In addition to drawing upon the recent published literature, we have developed some examples of applications based on our collective experiences in conducting ecological risk assessments.

Toxicity evaluation of trap and skeet shooting targets to aquatic test species.

: Large quantities of trap and skeet clay targets are used in shooting activities around the United States. For example, the number of targets used since 1970 has averaged approximately 560 million a year. A number of acute and chronic tests were performed to determine the toxicity of Remington Arms Company Blue Rock(®) trap and skeet target fragments upon selected freshwater and marine organisms. These studies were undertaken in support of an environmental impact study of trap and skeet shooting activities at a major gun club in the northeast United States. Targets were composed of approximately 67% dolomitic limestone, 32% petroleum pitch and 1% fluorescent aqueous paint (painted targets only). The majority of samples were painted, new targets obtained from the manufacturer and painted and aged targets collected around a shooting range. Additional tests were conducted using non-painted, new targets and leachates prepared from both painted, new and aged targets. Targets were crushed to small fragments and were either directly added to the test vessels at extremely high concentrations ranging from 670 to 600 000 mgl(-1) or used in leachate tests. In direct tests all target materials were essentially non-toxic to marine and freshwater organisms, except for the non-painted new targets which exhibited minimal acute toxicity to Daphnia magna (48 h EC 50=2200 mgl(-1)). In leachate tests, the leachate was not-toxic to mysid shrimp, the only organism tested. Additional samples of crushed targets were analysed for the presence of selected priority pollutants (EP toxicity test) and polycyclic aromatic hydrocarbons (PAHs). The targets did not exhibit the characteristics of toxicity as determined by the EP toxicity test but did contain substantial amounts of PAHs. However, results from new and aged targets suggest that PAH are tightly bound in the petroleum pitch and limestone matrix and are unlikely to be readily available in the environment. The potential impact of targets on the environment is further discussed.

Evaluation of benomyl and carbendazim in the in vivo aneuploidy/micronucleus assay in BDF1 mouse bone marrow.

Benomyl and its active metabolite carbendazim were investigated in BDF1 mouse bone marrow to establish whether micronuclei induced by these fungicides are caused by clastogenic or aneugenic events. Micronuclei were evaluated for kinetochores using immunofluorescent antikinetochore antibodies. Kinetochore positive (K+) micronuclei are likely to arise from chromosome loss since they presumably contain intact kinetochores and are indicative of aneuploidy. Conversely, kinetochore negative (K-) micronuclei are mostly likely to contain acentric chromosome fragments arising primarily from clastogenic damage. Benomyl and carbendazim were administered as single oral doses of 0.3, 8.6 or 17.2 mmol/kg (for benomyl, equivalent to 100, 2500 or 5000 mg/kg; for carbendazim, equivalent to 66, 1646 or 3293 mg/kg). Both compounds were positive in the micronucleus test at doses of 8.6 and 17.2 mmol/kg, and an average of 82% (benomyl) and 87% (carbendazim) of the total micronucleated polychromatic erythrocytes were K+. No effects were seen with either fungicide at 0.3 mmol/kg. These results are analogous to findings with known aneugens such as vincristine but are in contrast to results with classical clastogens such as cyclophosphamide. Thus, benomyl and carbendazim induce micronuclei in mouse bone marrow cells primarily through an aneugenic mechanism.

Activation and detection of (pro)mutagenic chemicals using recombinant strains of Streptomyces griseus.

Two recombinant strains of Streptomyces griseus have been developed to report on the activation of promutagenic chemicals. This activation is monitored by reversion of the bacterial test strains to a kanamycin-resistant phenotype. Strain H69 detects point mutations and was reverted at an increased frequency by acetonitrile, 2-aminoanthracene, 1,2-benzanthracene, benzidine, benzo(a)pyrene, 9,10-dimethyl-1,2-benzanthracene, and glycine. The second strain, FS2, detects frame shift mutations and was reverted at an increased frequency by 1,2-benzanthracene, benzidine, and glycine. Compounds such as butylated hydroxytoluene, catechol, chlorobenzene, hydroquinone, potassium chloride, phenol, cis-stilbene, trans-stilbene, and toluene did not elicit positive responses in either strain. In addition, these strains are capable of detecting direct-acting mutagens such as N-methyl-N'-nitrosoguanidine and ICR-191, providing further evidence of their promise for detecting a wider range of mutagens. To our knowledge, this is the first report of bacterial strains capable of activating promutagenic compounds and detecting their mutagenic metabolites without the benefit of an exogenous activation system such as the rodent liver homogenate (S9).

Application of the Hydra attenuata assay for identifying developmental hazards among natural waters and wastewaters.

This study concerns application of the Hydra attenuata assay to detect the developmental toxicity potential of various aqueous samples. First, the assay was modified for testing aqueous samples because water quality has a major impact on aquatic toxicity testing and the results thus obtained. Ranges of sample pH, salinity (conductivity), and hardness were examined for their adverse effects upon the hydra. Adult hydra were unaffected morphologically by pH 5.5-9.5, and the artificial embryo ("embryo") developed normally in a pH range of 6.25 to 8.25. For water hardness, the minimal affective concentration was 1000 mg/liter (as CaCO3) in adults and 625 mg/liter in the embryos; the NOAELs for these were 750 mg/liter in the adult and 250 mg/liter CaCO3 in the embryo. Salinity in excess of 5 ppt was lethal to adults and embryos, indicating the assay may not be applicable to marine or highly saline samples. Finally, grab samples were tested from rivers in Maryland, Pennsylvania, New Jersey, and Delaware, some of which are impacted by industrial and agricultural activities, as well as several samples of industrial wastewaters from one major facility. The assay functioned normally with these diverse samples and yielded results that can be used in assessing the potential developmental hazard of these materials.

The genetic toxicology of organic compounds in natural waters and wastewaters.

This review was drawn from the literature describing genotoxic organic compounds in natural water (rivers, lakes, streams) and wastewater, as well as from recent discussions with industrial scientists and environmental regulators. Testing of wastewaters for genotoxicity may become a routine requirement for some industrial wastewater discharge permits, not unlike the more common requirement for routine aquatic toxicity tests. The stimuli for this are concerns that aquatic organisms inhabiting waters impacted by wastewater discharges suffer an increased risk of genetic damage or cancer, and that humans utilizing these waters for recreational and drinking water purposes may suffer similar genetic or carcinogenic risks. Some evidence suggests that neoplasia in aquatic organisms is related to habitat contamination, yet field evaluations fail to substantiate adequately a cause-and-effect relationship. Because aquatic organisms respond like mammals to the same genotoxic compounds, the increased burden of genotoxic compounds to the environment may impact certain endemic species. Wastewater discharges may be one source of genotoxic organic compounds in those impacted areas. With respect to potential human health impacts, evidence is supportive of increased cancer risk to individuals drinking water from surface sources; however, this risk may or may not relate to whether the drinking water source received input of wastewater discharges or known carcinogens. Throughout the published literature reviewed herein, the Salmonella/Ames gene mutation test was widely used to assess genotoxic activity, although studies using indigenous plants and aquatic organisms as in vivo monitors of genotoxic activity exist. No "standard" or frequently followed protocols for sample collection, sample processing, selection of tests or their conduct, or interpretation of data exist for most of the genotoxicity studies reviewed. For the Salmonella/Ames test, the aqueous samples were concentrated usually on XAD resin or by liquid:liquid extraction, and without this concentration step few samples exhibited genotoxic activity. Hence, in most instances, the ambient concentration of the compounds causing this activity is below that which is readily detectable by this test, a finding not new to this review. In contrast, aquatic organisms in laboratory and field studies responded to ambient concentrations of genotoxic compounds, thus alleviating the need for sample concentration. However, there appears to be a reluctance to utilize this information for extrapolation to potential human health effects. Unfortunately, no generally accepted and scientifically validated protocol for preparing aqueous samples for genotoxicity testing exists. Developing such a protocol is necessary before embarking on widespread genotoxicity testing of wastewaters, especially if results are to be used for permit compliance.(ABSTRACT TRUNCATED AT 400 WORDS)

Activation of promutagenic chemicals by Streptomyces griseus containing cytochrome P-450soy.

Streptomyces griseus cells containing cytochrome P-450soy oxidize a diverse array of xenobiotic compounds. This metabolic capability was exploited for activation of promutagenic chemicals such as polycyclic aromatic hydrocarbons, aromatic amines and small aliphatics in a modified Salmonella/Ames plate incorporation assay using tester strains TA98 and TA1538. In this assay promutagens such as 3,3'-dimethylbenzidine, 3,3'-dimethoxybenzidine, benzidine, 2-acetylaminofluorene, 2-aminoanthracene, 2,4-diaminotoluene, 4-aminobiphenyl, benzo(a)pyrene, chloropicrin and N-nitrosodimethylamine were oxidized to mutagenic metabolites by S. griseus intact cells which mutated Salmonella tester strains (TA98 and TA1538). S. griseus failed to activate 7,12-dimethylbenzanthracene and 4-chloro-2-nitroaniline. In parallel tests performed with rat liver homogenate (S9), N-nitrosodimethylamine was not activated.

Mutagenic and cytogenetic analyses of organic extracts of simulated runoffs from model coal piles.

The use of coal as a fuel in utility and other industries in the United States is increasing. Typically, these utilities store their coal outdoors in large piles, and rainfall on the piles produces a runoff containing hazardous inorganic and organic materials. Four coals of varying sulfur contents, all used for fuel in the United States, were tested. Organic materials were extracted from simulated runoffs of model coal piles and were tested for mutagenicity with a Salmonella/microsomal assay and for clastogenicity and cytotoxicity in Chinese hamster ovary cells. The extracts of the high-sulfur coals and the lignite were more mutagenic and clastogenic than extracts from the low-sulfur coal.