Biomarker responses and disease susceptibility in juvenile rainbow trout Oncorhynchus mykiss fed a high molecular weight PAH mixture.

Juvenile rainbow trout were fed a diet containing an environmentally relevant mixture of 10 high molecular weight polycyclic aromatic hydrocarbons (PAHs) at a dose of 0.66 or 7.82 µg PAH · g fish(-1) · d(-1). At 3, 7, 14, and 28 d, biomarkers of aryl hydrocarbon receptor activation (AHR), hepatic microsomal ethoxyresorufin-O-deethylase (EROD) activity, and cytochrome P4501A (CYP1A)-associated staining increased 14- to 26-fold and 6- to 14-fold, respectively, in fish fed 7.82 µg PAH · g fish (-1) · d(-1). Cytochrome P4501A-associated staining increased 2- to 9-fold on days 3, 7, and 28 in fish fed 0.66 µg PAH · g fish(-1) · d(-1). Bile fluorescent aromatic compounds served as a biomarker of exposure and confirmed that PAH exposure was consistent over 50 d. DNA damage in blood cells, protein oxidation, and lipid peroxidation in the kidney were biomarkers of oxidative stress and all increased in fish fed 7.82 µg PAH · g fish(-1) · d(-1). Fish fed 0.66 µg PAH · g fish(-1) · d(-1) had elevated DNA damage in blood cells but increased protein oxidation or lipid peroxidation in the kidney were not observed. Challenge with Aeromonas salmonicida, at lethal concentration (LC) 20, decreased survival in fish previously fed either 0.66 µg PAH · g fish(-1) · d(-1) or 7.82 µg PAH · g fish(-1) · d(-1) relative to fish fed the control diet. In general, biomarkers of both AHR activation and oxidative stress peaked at 3 to 14 d then declined at 28 to 50 d of PAH exposure and an increase in susceptibility to disease was observed at 50 d. These results link PAH exposure to biomarker responses that may be useful as early indicators of population level responses, such as mortality resulting from an increase in disease susceptibility.

Advancing environmental toxicology through chemical dosimetry: external exposures versus tissue residues.

The tissue residue dose concept has been used, although in a limited manner, in environmental toxicology for more than 100 y. This review outlines the history of this approach and the technical background for organic chemicals and metals. Although the toxicity of both can be explained in tissue residue terms, the relationship between external exposure concentration, body and/or tissues dose surrogates, and the effective internal dose at the sites of toxic action tends to be more complex for metals. Various issues and current limitations related to research and regulatory applications are also examined. It is clear that the tissue residue approach (TRA) should be an integral component in future efforts to enhance the generation, understanding, and utility of toxicity testing data, both in the laboratory and in the field. To accomplish these goals, several key areas need to be addressed: 1) development of a risk-based interpretive framework linking toxicology and ecology at multiple levels of biological organization and incorporating organism-based dose metrics; 2) a broadly applicable, generally accepted classification scheme for modes/mechanisms of toxic action with explicit consideration of residue information to improve both single chemical and mixture toxicity data interpretation and regulatory risk assessment; 3) toxicity testing protocols updated to ensure collection of adequate residue information, along with toxicokinetics and toxicodynamics information, based on explicitly defined toxicological models accompanied by toxicological model validation; 4) continued development of residue-effect databases is needed ensure their ongoing utility; and 5) regulatory guidance incorporating residue-based testing and interpretation approaches, essential in various jurisdictions.

Impaired growth in the polychaete Armandia brevis exposed to tributyltin in sediment.

Juveniles of the opheliid polychaete, Armandia brevis, were exposed to sediment-associated tributyltin (TBT) for 42 days to evaluate toxicity and bioaccumulation. Growth in this species was inhibited in a dose-response fashion by increasing concentrations of TBT. Even though the biota-sediment accumulation factor (BSAF) for TBT declined for the higher sediment concentrations, the total butyltins in tissue increased over all sediment concentrations. At the highest sediment concentrations, polychaetes bioaccumulated less TBT than expected, which was most likely due to reduced uptake and continued metabolism of the parent compound. The less than expected BSAF values exhibited by animals at the exposure concentrations causing severe effects are an important finding for assessing responses in the field. It appears that severe biological effects can occur in long-term experiments without the expected high tissue concentrations; an observation likely explained by altered toxicokinetics. Analysis of variance determined the lowest observed effect concentration for growth to be 191 ng/g sediment dry wt. for 21 days of exposure and 101 ng/g sediment dry wt. at day 42, indicating that 21 days was insufficient for delineating the steady-state toxicity response. When based on regression analysis, the sediment concentration causing a 25% inhibition in growth at 42 days exposure was 93 ng/g dry wt. (total organic carbon = 0.58%). A dose-response association was also determined for polychaete net weight and TBT in tissue. The tissue residue associated with a 25% reduction in growth was 2834 ng/g dry wt. at day 42. A comparison of these results with previous work indicates that juveniles are approximately three times more sensitive than adults to TBT exposure. The sediment concentrations affecting growth in this species are commonly found in urban waterways indicating potentially severe impacts for this and other sensitive species.

Predicting the fate and effects of tributyltin in marine systems.

The available data indicate that sediment-water partitioning, bioaccumulation, and the toxicity responses for tributyltin (TBT) are predictable when using some of the assumptions and tenets of the equilibrium partitioning method, toxicokinetic modeling (1CFOK), and critical body residue (CBR) approach. Because TBT is ionizable, its speciation is strongly affected by pH, which appears to cause large variations in the octanol-water partition coefficient. In marine systems, and in freshwater systems with high pH, TBT occurs predominantly in the hydroxide form, which may explain the hydrophobic properties and its EqP behavior. Organic carbon in sediment (> 0.2%) appears to be the major controlling factor for sediment-water partitioning. The equilibrium organic carbon-normalized sediment-water partition coefficient (Koc) in marine environments is approximately 32,000 (log10 Koc approximately 4.5), which was determined from direct measurement and confirmed by the relationship between the lipid-normalized bioconcentration factor (BCF) in porewater and the biota-sediment accumulation factor (BSAF). The conclusion that sediment-water partitioning of TBT in marine systems follows EqP is supported by the similarity between its Kow and Koc and the correlation between the sediment-water partition coefficient (Kp) and sediment TOC, which results from the influence of organic carbon on pore-water concentrations. Even though the rates of uptake and elimination control tissue residues and lipid content appears to have no bearing on the amount of TBT that is accumulated, the species specific BSAF is useful for examining bioaccumulation, sediment-water partitioning, and the toxicity response. Although TBT is hydrophobic and appears to have a propensity to accumulate in lipid, the rates of uptake and elimination, not thermodynamics, appear to control whole-body tissue concentrations. Support for a toxicokinetic approach for predicting tissue residues is found in BCF and BSAF values for several species that are far in excess of that predicted by simple thermodynamic partitioning and in the comparisons of observed and predicted bioaccumulation values based on toxicokinetic coefficients. This observation is counter to the assumption of EqP that the route of uptake is of no consequence under equilibrium conditions. For TBT, it appears that kinetics determine tissue residues and that body lipid is important only for regulating the toxic response, not the amount bioaccumulated. Unlike those for neutral hydrophobic organic compounds, the toxicokinetics for this one toxicant are highly variable in diverse species but relatively accurate in predicting the amount bioaccumulated and the resulting toxicity response. For the CBR approach to be useful, a relatively constant tissue residue for a given biological response is necessary. Several studies support the CBR approach because certain biological effects, such as mortality and growth inhibition, occur at a relatively constant TBT tissue concentration. For TBT, the lethal whole-body tissue concentration affecting 50% of individuals (LR50) exhibits little variation, occurring at approximately 48 micrograms/g (166 nmol/g) dry weight in a range of species. Direct evidence and correlation of the LC50 and the bioconcentration factor (BCF) support this observation. Impaired growth, a sublethal response, also appears to be associated with a relatively constant tissue concentration, which has also been demonstrated by direct measurement and indirectly by regression of the BCF and LOEC. The lowest-observed-effect tissue residue (LOER) associated with impaired growth for several species was approximately 3 micrograms/g (10.4 nmol/g) dry wt. Because of the small number of studies linking growth impairment and tissue concentrations, additional studies are needed to confirm these values. (ABSTRACT TRUNCATED)

Organochlorines in stranded pilot whales (Globicephala melaena) from the coast of Massachusetts.

Pilot whales strand periodically along the U.S. coast, and these strandings offer an opportunity for the collection of tissues for biomonitoring of contaminant exposure in cetaceans, as well as for specimen archiving. Concentrations of organochlorine (OC) contaminants (e.g., PCB congeners, pesticides, DDTs) were measured in tissue samples from pilot whales that stranded in 1986 and 1990 along the Massachusetts coast. Adult and fetal samples of blubber, liver, brain, and kidney were collected, as well as ovaries from mature female whales. Many of the OCs found in maternal tissues were detected in corresponding fetal tissues indicating maternal transfer of OCs to the fetus. The concentrations of individual OCs in tissues varied considerably among the animals. Statistically significant differences were found between females and males for the concentrations of certain analytes (e.g., SigmaPCBs, p,p'-DDE) and these differences may be partially due to contaminants being transferred by the female whales during gestation and lactation. The concentrations of OCs in different tissues were similar when based on total lipid weight, except for the brain, which contained the lowest lipid-normalized OC concentrations. The low concentrations in brain may be related to the disparate lipid compositions in this tissue as well as the presence of the blood-brain barrier. The availability of data on these archived and biomonitoring samples provides a baseline for future retrospective studies.

Comparison of elements in bottlenose dolphins stranded on the beaches of Texas and Florida in the Gulf of Mexico over a one-year period.

We analyzed tissue samples from bottlenose dolphins (Tursiops truncatus) that had stranded on beaches in Texas and Florida over a 1-year period starting in September 1991. The concentrations of 10 elements plus methyl mercury (MeHg) were determined in brain, kidney, and liver, and we examined these results for differences based upon age, site, sex, and tissue type. A strong inverse relationship between total mercury (Hg) and the percentage that was MeHg was found in liver, kidney, and brain tissue, presumably due to demethylation of MeHg. A threshold concentration was found for total Hg in brain tissue, indicating that most Hg was present as MeHg up to about 8 years of age. Increases in total Hg after this age were accompanied by an increase in the ratio of total Hg to MeHg, indicating demethylation. Strong relationships were found between total Hg in liver and age and between total Hg and selenium in liver, which have been observed before in many fish- and squid-eating marine mammals. The only difference based on sex of the animals was observed for MeHg, which was higher in females and contrary to the pattern often observed for organic contaminants. Several elements (copper, Hg, lead, zinc) exhibited intersite differences, which were not consistent. Bottlenose dolphin from Florida exhibited the highest levels of MeHg and total Hg, while animals from Texas exhibited the highest levels of lead, copper, and zinc. The essential elements copper and zinc were expected to be the same for the Texas and Florida animals; however, observed differences may indicate population differences in basic physiological levels, dietary intake, or health status.

Chemical contaminants in harbor porpoise (Phocoena phocoena) from the north Atlantic coast: tissue concentrations and intra- and inter-organ distribution.

Concentrations of chlorinated hydrocarbons (CHs), such as polychlorinated biphenyls (PCBs), were measured in subsamples taken from different anatomical locations of blubber and liver of three apparently healthy harbor porpoises (Phocoena phocoena) incidentally caught in a gill-net fishery along the northwest Atlantic coast; selected elements (e.g., mercury) were measured in subsamples of liver. The vertical distribution (skin to muscle) of contaminants within blubber was also determined. Additionally, the concentrations of CHs and elements were determined in individual samples of brain, lung, kidney, and testis to assess how the disposition of toxic chemicals may be dependent on the physiological characteristics of a specific organ. Statistical analyses of the results showed that the anatomical location of the blubber or liver sample had no significant effect on concentrations of either CHs in blubber and liver, or of selected elements in liver. However, there were statistical differences between strata of blubber (skin to muscle) for the concentrations of CHs. As expected, the results showed that the CH concentrations, based on wet weight, were considerably higher in the blubber than in the other tissues; however, the concentrations of CHs in the different tissues were more comparable when values were based on total lipid weight with the exception of the brain where lipid normalized concentrations were lower than in all other tissues: This low relative accumulation of lipophilic contaminants in the brain tissue may be due to the presence of the blood-brain barrier, or due to a lower proportion of neutral lipids, such as triglycerides, as analysis for percent lipid and for the proportion of specific lipid classes showed.

Comparative bioaccumulation of chlorinated hydrocarbons from sediment by two infaunal invertebrates.

Bioaccumulation of chlorinated hydrocarbons (CHs) from field-contaminated sediments by two infaunal invertebrates, Rhepoxynius abronius (a non-deposit feeding amphipod) and Armandia brevis (a nonselective, deposit-feeding polychaete), was examined and species responses were compared. Sediments were selected over a large geographical area of the Hudson-Raritan estuary to assess the potential for bioaccumulation from a typical urban estuary. Unlike polycyclic aromatic hydrocarbons (PAHs) from these sediments, concentrations of CHs in interstitial water (IW) indicated that partition coefficients (Koc) were generally as expected, especially when based on predicted, nonsorbed, interstitial water CH concentrations (IWfree). Correlations between amphipod and polychaete tissue residues revealed that these species were responding similarly to a gradient of CH concentrations in sediment. While tissue residues and BAFloc (lipid/organic carbon normalized bioaccumulation factor) values for the trichlorobiphenyls were similar for both species, accumulation in the polychaete was three to 10 times higher for the more hydrophobic PCBs, which was attributed to differences in the route of exposure. A negative correlation between the bioaccumulation factor (BAF) and total organic carbon (TOC) was found for both species, which was expected according to equilibrium partitioning theory. Because it was assumed that the amphipod was not feeding in these tests and the polychaete was ingesting sediment, comparison of their tissue residues and bioaccumulation factors was useful for highlighting the importance of sediment ingestion, especially for short-term, nonequilibrium exposures. These results may also help elucidate the limitations associated with assessing bioaccumulation and the resultant toxic response in standard 10-day toxicity tests with similar invertebrates.

Bioaccumulation of polycyclic aromatic hydrocarbons by marine organisms.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the marine environment, occurring at their highest environmental concentrations around urban centers. While they can occur naturally, the highest concentrations are mainly from human activities, and the primary sources are combustion products and petroleum. Two factors, lipid and organic carbon, control to a large extent the partitioning behavior of PAHs in sediment, water, and tissue; the more hydrophobic a compound, the greater the partitioning to these phases. These two factors, along with the octanol-water partition coefficient, are the best predictors of this partitioning and can be used to determine PAH behavior and its bioavailability in the environment. It is well known that the lipid of organisms contains the highest levels of hydrophobic compounds such as PAHs, and that organic carbon associated with sediment or dissolved in water can have the greatest influence on PAH bioavailability. Partitioning of combustion-derived PAHs between water and sediment may be much less than predicted, possibly because associations with particles are much stronger than expected. This reduced partitioning may produce erroneous results in predicting bioaccumulation where uptake from water is important. Accumulation of PAHs occurs in all marine organisms; however, there is a wide range in tissue concentrations from variable environmental concentrations, level and time of exposure, and species ability to metabolize these compounds. PAHs generally partition into lipid-rich tissues, and their metabolites can be found in most tissues. In fish, liver and bile accumulate the highest levels of parent PAH and metabolites; hence, these are the best tissues to analyze when determining PAH exposure. In invertebrates, the highest concentrations can be found in the internal organs, such as the hepatopancreas, and tissue concentrations appear to follow seasonal cycles, which may be related to variations in lipid content or spawning cycles. The major route of uptake for PAHs has been debated for years. For the more water-soluble PAHs, it is believed that the main route of uptake is through ventilated water and that the more hydrophobic compounds are taken in mainly through ingestion of food or sediment. There are many variables, such as chemical hydrophobicity, uptake efficiency, feeding rate, and ventilatory volume, which may affect the outcome. The route of uptake may be an important issue for short-term events; however, under long-term exposure and equilibrium conditions between water, prey, and sediment, the route of uptake may be immaterial because the same tissue burdens will be achieved regardless of uptake routes.(ABSTRACT TRUNCATED AT 400 WORDS)

Chemical contaminants in gray whales (Eschrichtius robustus) stranded along the west coast of North America.

The concentrations of selected chlorinated hydrocarbons (e.g. PCBs, DDTs, DDEs, chlordanes) and essential (e.g. zinc, selenium, copper) and toxic (e.g. mercury, lead, arsenic) elements were measured in tissues and stomach contents from 22 gray whales (Eschrichtius robustus) stranded between 1988 and 1991 at sites from the relatively pristine areas of Kodiak Island, AK, to more urbanized areas in Puget Sound, WA, and San Francisco Bay, CA. The majority of animals were stranded at sites on the Washington outer coast and in Puget Sound. The gray whale has the unique feeding strategy among Mysticeti of filtering sediments to feed on benthic (bottom dwelling) invertebrates. Thus, the wide geographical distribution of the stranded whales allowed (1) an initial assessment of whether concentrations of chemical contaminants in these whales exhibited region specific differences and (2) whether toxic chemicals that accumulate in sediments may have contributed to the mortality and stranding of gray whales near the more polluted urban areas. Analyses for chlorinated hydrocarbons in blubber from 22 animals showed no apparent significant differences among stranding sites. The concentrations of sigma PCBs and sigma DDEs in blubber, for example, ranged from 120 to 10,000 and 9 to 2100 p.p.b. (ng/g) wet weight, respectively. Additionally, analyses of chlorinated hydrocarbons and selected elements in liver (n = 10) also showed no apparent significant differences between whales stranded in Puget Sound and whales stranded at more pristine sites (Alaska, Washington outer coast and Strait of Juan de Fuca and Strait of Georgia). For example, the concentrations of sigma PCBs and sigma DDEs in liver ranged from 79 to 1600 and 7 to 280 p.p.b., respectively, and the concentrations of the toxic elements, mercury and lead ranged from 9 to 120 and 20 to 270 p.p.b., respectively. Analyses of stomach contents revealed low concentrations of chlorinated hydrocarbons, but high concentrations (wet weight) of aluminum (1,700,000 +/- 450,000 p.p.b.), iron (320,000 +/- 250,000 p.p.b.), manganese (23,000 +/- 15,000 p.p.b.), and chromium (3400 +/- 1300 p.p.b.), but no significant differences were observed between whales stranded in Puget Sound compared to whales stranded at the more pristine sites. The relative proportions of these elements in stomach contents of stranded whales were similar to the relative proportions in sediments, which is consistent with a geological source of these elements from the ingestion of sediment during feeding. Thus, overall, the concentrations of anthropogenic chemicals in stranded gray whales showed little relation to the levels of chemical contaminants at the stranding sites.(ABSTRACT TRUNCATED AT 400 WORDS)