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.

Biomarkers of contaminant exposure in Northern Pike (Esox lucius) from the Yukon River Basin, Alaska.

As part of a larger investigation, northern pike (n = 158; Esox lucius) were collected from ten sites in the Yukon River Basin (YRB), Alaska, to document biomarkers and their correlations with organochlorine pesticide (total p,p'-DDT, total chlordane, dieldrin, and toxaphene), total polychlorinated biphenyls (PCBs), and elemental contaminant (arsenic, cadmium, copper, lead, total mercury, selenium, and zinc) concentrations. A suite of biomarkers including somatic indices, hepatic 7-ethoxyresorufin O-deethylase (EROD) activity, vitellogenin concentrations, steroid hormone (17B- ustradiol and 16-kebtestosteront) concentrations, splenic macrophage aggregates (MAs), oocyte atresia, and other microscopic anomalies in various tissues were documented in YRB pike. Mean condition factor (0.50 to 0.68), hepatosomatic index (1.00% to 3.56%), and splenosomatic index (0.09% to 0.18%) were not anomalous at any site nor correlated with any contaminant concentration. Mean EROD activity (0.71 to 17.51 pmol/min/mg protein) was similar to basal activity levels previously measured in pike and was positively correlated with selenium concentrations (r = 0.88, P < 0.01). Vitellogenin concentrations in female (0.09 to 5.32 mg/mL) and male (<0.0005 to 0.097 mg/mL) pike were not correlated with any contaminant, but vitellogenin concentrations >0.01 mg/mL in male pike from multiple sites indicated exposure to estrogenic compounds. Mean steroid hormone concentrations and percent oocyte atresia were not anomalous in pike from any YRB site. Few site differences were significant for mean MA density (1.86 to 6.42 MA/mm(2)), size (812 to 1481 microm(2)), and tissue occupied (MA-%; 0.24% to 0.75%). A linear regression between MA-% and total PCBs was significant, although PCB concentrations were generally low in YRB pike (< or =63 ng/g), and MA-% values in female pike (0.24% to 0.54%) were lower than in male pike (0.32% to 0.75%) at similar PCB concentrations. Greater numbers of MAs were found as zinc concentrations increased in YRB female pike, but it is unlikely that this is a causative relationship. Histological abnormalities observed in gill, liver, spleen, and kidney tissues were not likely a result of contaminant exposure but provide information on the general health of YRB pike. The most common histologic anomalies were parasitic infestations in various organs and developing nephrons and nephrocalcinosis in posterior kidney tissues. Overall, few biomarker responses in YRB pike were correlated with chemical contaminant concentrations, and YRB pike generally appeared to be healthy with no site having multiple anomalous biomarker responses.

Histopathology of feral fish from a PCB-contaminated freshwater lake.

The purpose of this study was to evaluate the potential toxic effects of chronic sublethal polychlorinated biphenyl (PCB) exposure on feral fish, using histopathology as an endpoint. Histopathological study of bream (Abramis brama) and asp (Aspius aspius) living in a PCB-polluted freshwater lake revealed abnormal cellular changes in the renal corpuscle of both species. Dilation of glomerular capillaries (DGC), mesangial edema (ME), an adhesion between visceral and parietal layers of Bowman's capsule (ABC), and filling of Bowman's space (FBS), were highly prevalent features in lake fish. The prevalence of each of these lesions was significantly lower, or totally absent in fish caught from reference locations. Cellular alterations in liver, gill, gonads, spleen, and intestine were all linked to seasonal changes. The results suggest that some of the observed histopathological changes in renal glomeruli, particularly DGC and ME, could possibly indicate a prolonged chemical stress caused by PCBs and related compounds. It is also possible that chronic PCB exposure may have suppressed and weakened the immuno systems of exposed fish making them more vulnerable to secondary parasitic infections.

Molecular detection and phylogenetic placement of a microsporidian from English sole (Pleuronectes vetulus) affected by X-cell pseudotumors.

Flatfish tissue samples exhibiting X-cell pseudotumors were tested with a number of ribosomal DNA (rDNA) general primers in polymerase chain reactions (PCRs). Microsporidian primers resulted in the amplification of an rDNA fragment and molecular phylogenetic analysis indicated that although the organism did not relate closely with any current microsporidian genera, it was most similar to Nucleospora salmonis and branched within the Enterocytozoonidae. Re-examination of the original tissues used for DNA extractions revealed the presence of putative microsporidian spores in PCR-positive samples. These observations reiterate the highly sensitive diagnostic feature of PCR, allowing detection of organisms overlooked by conventional methods and demonstrate the occurrence of rare, coinfecting organisms.

Hepatic lesions in a redstriped rockfish (Sebastes proriger) suggestive of a herpesvirus infection.

Unusual lesions were observed in a redstriped rockfish (Sebastes proriger) collected during a survey of marine fishes off the coast of British Columbia, Canada. This particular fish exhibited prominent hepatomegaly, with large, coalescing, multiple hemorrhages. The affected liver exhibited remarkable histological changes that, taken together, strongly suggested infection by a virus of the herpesvirus group. Multiple, multinucleated giant cells or syncytia of hepatocytes occurred throughout the liver and were associated with massive, coalescing areas of coagulation necrosis, edema, congestion and cavernous hemorrhages (peliosis hepatis) with thrombosis. In addition, the liver showed multifocal inflammation, characterized by perivascular and peribiliary cuffing of mononuclear inflammatory cells. High magnification of the syncytia revealed that the nuclei were pleomorphic, hyperchromatic, and typically contained eosinophilic to densely amphophilic inclusion bodies of varying size, closely resembling Cowdry Type A inclusions. These inclusions stained red to purple in Feulgen's stain, indicating presence of DNA. Electron-lucent spheres (approximately 100 nm diameter) were observed within hepatocyte nuclei by transmission electron microscopy, suggestive of herpesvirus capsids. To our knowledge, this is the first report of a putative or confirmed herpesvirus infection in any rockfish of the genus Sebastes.

From sediment bioassay to fish biomarker--connecting the dots using simple trophic relationships.

Two common problems in applying and interpreting invertebrate bioassays and fish biomarkers in sediment toxicology are the wide gap between significant effects concentrations determined by these two approaches, and a general lack of ecological context. We have devised an exposure system that is able to reconcile much of the disparity between invertebrate bioassay and fish biomarker results by incorporating realistic ecological processes based on deposit feeding and predator-prey interactions. This system relates the disturbance of interest (sediment contamination) to biologically meaningful effects in a resource of interest (marine flatfish) via a realistic contaminant vector (a deposit-feeding polychaete worm). In this pilot study, polychaetes (Armandia brevis) were exposed for 28 days to clean sediments supplemented with benzo(a)pyrene (BaP), para-para dichlorodiphenyldichloroethylene (pp'DDE), Aroclor 1254, or field sediments collected from two sites in Puget Sound, Washington, contaminated predominantly with polcyclic aromatic hydrocarbons (PAHs) or chlorinated compounds. Exposed worms were then fed live to juvenile English sole (Pleuronectes vetulus) for 10 or 12 days. At the end of the exposure period, fish were measured for length and weight, sacrificed, and preserved for either routine histopathology and immunohistochemical analysis of cytochrome P450 1A induction, or 32P post-labeling determination of hepatic PAH-DNA adducts. Growth of predatory flatfish was lower than reference in all but one of eight groups fed contaminant-exposed polychaetes; however, statistically significant reductions in growth were only observed in three of these eight groups, at least in part due to low statistical power. Juvenile sole from all contaminant-exposed groups showed increased expression of CYP1A, and fish exposed to BaP-exposed worms showed clear evidence of hepatic PAH-DNA adducts. This method allows the concurrent evaluation of sediment contamination at multiple biological and ecological levels. These results indicate that sediments determined to be nontoxic by common invertebrate bioassays may have the potential to cause adverse effects at higher trophic levels.

Molecular epizootiology of genotoxic events in marine fish: linking contaminant exposure, DNA damage, and tissue-level alterations.

Molecular epizootiological studies are increasingly being used to investigate environmental effects of genotoxic contaminants. The assessment of damage to DNA and linking the damage to subsequent molecular, cellular, or tissue-level alterations is a central component of such studies. Our research has focused on the refinement of the 32P-postlabeling assay for measuring covalent DNA-xenobiotic adducts arising from exposure to polycyclic aromatic compounds, using DNA adducts as molecular dosimeters of genotoxic contaminant exposure in biomonitoring studies, and investigating the relationship of DNA adduct formation to toxicopathic liver disease, including neoplastic lesions. A combination of field and laboratory studies using the 32P-postlabeling assay has shown that DNA adducts in marine fish are effective molecular dosimeters of genotoxic contaminant exposure. Investigations of the relationship of DNA adduct formation to neoplastic liver disease have shown that elevated levels of DNA adducts in certain fish species from contaminated coastal sites are associated with increased prevalences of toxicopathic hepatic lesions, including neoplasms, and that the ability to assess DNA damage has helped to explain, in part, species differences in lesion prevalence. Moreover, in a study of a site in Puget Sound contaminated with polycyclic aromatic compounds, we have shown, for the first time, that elevated levels of hepatic DNA adducts are a significant risk factor for certain degenerative and preneoplastic lesions occurring early in the histogenesis of hepatic neoplasms in feral English sole (Pleuronectes vetulus). These latter findings coupled with our current studies of mutational events in the K-ras proto-oncogene should provide further mechanistic substantiation that mutagenic events resulting from exposure to complex mixtures of genotoxic polycyclic aromatic compounds are involved in the etiology of hepatic neoplasia in English sole.

Hydropic vacuolation in the liver of three species of fish from the U.S. West Coast: lesion description and risk assessment associated with contaminant exposure.

Hydropic vacuolation (HydVac) of biliary epithelial cells and hepatocytes is described for 3 species of U.S. West Coast bottom fishes. Risk assessment analyses are also conducted to determine if the prevalence of this lesion increases in association with contaminant exposure and site of capture. The morphology of HydVac in starry flounder Platichthys stellatus, white croaker Genyonemus lineatus and rock sole Lepidopsetta bilineata was similar to that described in winter flounder Pleuronectes americanus from the U.S. Atlantic Coast, especially in that HydVac most commonly affected biliary epithelial cells. Hydropic vacuolation was the most prevalent liver lesion in starry flounder and white croaker captured from contaminated environments. Risk assessment analyses confirmed that the relative risk for HydVac increased with the presence of aromatic and chlorinated hydrocarbons in sediment, fish bile, and fish liver for these species. Hydropic vacuolation also frequently occurred in rock sole, but the lesion showed no clear association with contaminant exposure in this species. The types of liver lesions that were useful biomarkers of contaminant effects in fish depended on the species and this factor must be taken into account when evaluating histopathological biomarkers of response to contaminant exposure.

Immunohistochemical localization of CYP1A and CYP3A-like isozymes in hepatic and extrahepatic tissues of Atlantic cod (Gadus morhua L.), a marine fish.

The cellular localization of inducible CYP1A and constitutive CYP3A-like forms in different organ systems of Atlantic cod (Gadus morhua) was determined in control fish and fish exposed to beta-naphthoflavone (BNF). Paraffin-embedded sections were stained with polyclonal rabbit anti-cod P450 1A IgG or rabbit anti-rainbow trout P450con (a putative CYP3A form which cross-reacts with purified cod P450b) serum by the avidin-biotin peroxidase complex method. Following BNF-exposure of cod, CYP1A induction was immunohistochemically demonstrated in hepatocytes and endothelial cells of liver, the endocardium and vascular endothelium in the atrium and ventricle, and epithelial cells of the proximal tubular segment, endothelial cells, and interrenal cells in kidney. The vascular endothelium was the main site of induction of CYP1A in gills, spleen, gut, pyloric caecae, and gonads. The CYP3A-like isozyme P450b was mainly localized to hepatocytes, renal tubular epithelium, and epithelial cells of the mucosa in the intestine. Furthermore, the distribution of P450b was not affected by BNF exposure. The localization of P450b bears interesting similarities to the localization of CYP3A in mammals supporting the CYP3A-like identity of cod P450b. Simultaneous localization of inducible CYP1A and a constitutively expressed CYP isoenzyme has not previously been reported in fish. This is also the first presentation of cellular distribution of a CYP3-like isozyme in fish. Staining of CYP1A in endothelial cells supports previous observations that endothelium is a major site of CYP1A induction following xenobiotic exposure in fish. The observation of CYP1A induction in interrenal cells has important implications for possible endocrine effects of xenobiotic exposure.

Relationships between toxicopathic hepatic lesions and exposure to chemical contaminants in English sole (Pleuronectes vetulus), starry flounder (Platichthys stellatus), and white croaker (Genyonemus lineatus) from selected marine sites on the Pacific Coast, USA.

Relationships between hepatic lesions and chemical contaminant concentrations in sediments, stomach contents, liver tissue, and bile were statistically evaluated in three species of bottomfish, English sole (Pleuronectes vetulus), starry flounder (Platichthys stellatus), and white croaker (Genyonemus lineatus), captured from 27 urban and nonurban sites on the Pacific Coast from Alaska to southern California. Lesions detected were neoplasms, preneoplastic foci of cellular alteration, nonneoplastic proliferative lesions, unique or specific degenerative/necrotic lesions, nonspecific degenerative/necrotic lesions, and hydropic vacuolation of biliary epithelial cells and hepatocytes. In general, lesion prevalences were significantly higher in all three species captured at chemically contaminated urban sites, and certain lesions had significantly higher relative risks of occurrence at urban sites in Puget Sound, San Francisco Bay, the vicinity of Los Angeles, and San Diego Bay. Concentrations of polycyclic aromatic hydrocarbons, polychlorinated biphenyls, DDT and its derivatives, and chlordanes and dieldrin in sediment, stomach contents, liver, and fluorescent aromatic compounds in bile were significant risk factors for the occurrence of neoplastic, preneoplastic, nonneoplastic proliferative, and specific degenerative/necrotic lesions, as well as hydropic vacuolation. Fish age also had a significant influence on occurrence of several hepatic lesions, but gender was rarely a significant risk factor. These relationships provide strong evidence for the involvement of environmental contaminants in the etiology of hepatic lesions in several marine bottomfish species and clearly indicate the utility of these lesions as biomarkers of contaminant-induced effects in wild fish.

Feasibility of postpartum rapid hospital discharge: a study from a community hospital population.

We have examined the safety and efficacy of an early postpartum discharge program in 289 patients who were identified prior to delivery and counseled and educated for discharge 12 to 36 hours after delivery. A total of 55% of the study patients were able to be discharged early. There were 4.3% significant maternal problems and 3% significant neonatal problems identified in the first 72 hours after delivery by a nurse practitioner home visit. Significant maternal problems after 72 hours (10%) and neonatal problems (5%) were also found by follow-up visits. The hospital readmission rate was 1.8%. These incidences are consistent with other studies in the literature and suggest that some type of early follow-up of both the mother and infant should routinely accompany early postpartum discharge programs.

Relationships between hepatic neoplasms and related lesions and exposure to toxic chemicals in marine fish from the U.S. West Coast.

English sole (Parophrys vetulus) inhabiting polluted waterways and embayments of Puget Sound, Washington, are affected with a variety of multiple, co-occurring idiopathic hepatic lesions, including unique degenerative conditions, putatively preneoplastic foci of cellular alteration, and neoplasms. Results of a statistical analysis of the patterns of co-occurrence of these lesions in wild English sole are consistent with the concept that these lesions represent morphologically identifiable steps forming a sequence of progression ultimately leading to the development of hepatic neoplasms. This progressive sequence parallels the pattern identified in experimental models of chemically induced hepatocarcinogenesis in rodents. The rationale for the hypothesis that these lesions in wild English sole can be caused by exposure to certain hepatoxic and hepatocarcinogenic xenobiotic compounds in the marine environment is based on the demonstration of significant and consistent statistical associations between levels of aromatic hydrocarbons (AHs) in sediment and prevalences of these idiopathic liver lesions; a significant contribution by sediment AHs to the variability in hepatic neoplasm prevalence in a logistic regression model; significantly increased probabilities for several idiopathic lesions in sole from chemically contaminated sites in Puget Sound; significant correlations between uptake of polycyclic aromatic hydrocarbons, as measured by levels of fluorescent metabolites of aromatic compounds in bile of sole, and prevalences of several hepatic lesion types; and experimental induction of unique degenerative, proliferative, and putatively preneoplastic focal lesions in English sole injected with either benzo(a)pyrene or a polycyclic aromatic hydrocarbons (PAH) enriched fraction of an extract from a contaminated urban sediment from Puget Sound.(ABSTRACT TRUNCATED AT 250 WORDS)

Consistent associations between hepatic lesions in English sole (Parophrys vetulus) and polycyclic aromatic hydrocarbons in bottom sediment.

A consistent and statistically significant association between prevalence of hepatic neoplasms in free-living sole (Parophrys vetulus) and levels of anthropogenic polycyclic aromatic hydrocarbons in bottom sediment from sites of fish capture was documented in a series of studies conducted over a period of 7 years in Puget Sound, Washington. This result strengthens the evidence supporting a causal relationship between exposure to sediment-associated hydrocarbons and development of hepatic neoplasms in this bottom-dwelling marine fish species. Prevalences of two other distinct categories of idiopathic hepatic lesions-megalocytic hepatosis and steatosis/hemosiderosis-also showed consistent, statistically significant associations with polycylic aromatic hydrocarbon concentrations in bottom sediment, and association with prevalence of a third category (putatively preneoplastic foci of cellular alteration) approached statistical significance. On the basis of other studies, megalocytic hepatosis and foci of cellular alteration are both considered to be important precursor lesions in the stepwise histogenesis of hepatic neoplasms.

Overview of studies on liver carcinogenesis in English sole from Puget Sound; evidence for a xenobiotic chemical etiology. I: Pathology and epizootiology.

Livers of wild English sole (Parophrys vetulus) from polluted waterways and embayments of Puget Sound, Washington, are affected by a spectrum of multiple, co-occurring idiopathic hepatic lesions, including neoplasms, putative preneoplastic foci of cellular alteration, and unique degeneration conditions. Results from a statistical analysis of the patterns of co-occurrence of these lesions in wild English sole indicate that these lesions represent morphologically identifiable steps leading to the development of hepatic neoplasms. This sequence parallels the lesion progression in experimental models of chemically induced liver carcinogenesis in rodents. The hypothesis that these lesions in wild English sole can be caused by exposure to certain xenobiotic hepatotoxic and hepatocarcinogenic compounds in Puget Sound is based on: a) statistical associations between levels of aromatic hydrocarbons (sigma AHs) in sediment and prevalences of these idiopathic liver lesions, b) the contribution of sigma AHs in accounting for the variability in hepatic neoplasm prevalence in a logistic regression model, c) elevated odds ratios for several idiopathic hepatic lesion types in sole from polluted sites in Puget Sound, d) significant correlations between prevalences of idiopathic hepatic lesions and levels of fluorescent metabolites of aromatic compounds (FACs) in bile of English sole, and e) experimental induction of putatively preneoplastic focal lesions in English sole injected with a PAH-enriched fraction of an extract from a contaminated urban sediment from Puget Sound, that were morphologically identical to lesions found in wild English sole from the same site.

The ultrastructure and histology of cholangiocellular carcinomas in English sole (Parophrys vetulus) from Puget Sound, Washington.

The ultrastructure and histology of cholangiocellular carcinomas from feral English sole (Parophyrs vetulus) living in polluted waterways of Puget Sound, WA. are described. Electron microscopy confirmed that biliary epithelial cells were the main proliferative cell type composing this variety of neoplasm. The arrangement of these cells varied from well-organized multiple bile duct-like structures to disorganized multilayered sheets of poorly differentiated biliary epithelial cells. A fibrous stroma consisting of multiple layers of collagen fibers and fibroblasts, with macrophages and various blood cell types scattered among these layers occurred between bile duct-like structures or aggregates of biliary epithelial cells. Hepatocytes were not apparent in these neoplasms except within small necrotic regions surrounded by neoplastic biliary epithelial cells. No virus-like particles were observed among the cases examined in this study.

Effect of bromobenzene and O-bromophenol on kidney and liver of English sole (Parophrys vetulus).

1. English sole (Parophrys vetulus) were injected intraperitoneally with a single dose of 9.8 mmol bromobenzene/kg of fish or 1.9 mmol O-bromophenol/kg of fish, both known renal toxicants in mammals. 2. Kidney, liver, gill spleen, intestines, heart and blood samples were subsequently obtained up to 48 hr post-injection for determination of microscopic lesions, concentrations of selected tissue antioxidants (glutathione and ascorbic acid), and selected serum parameters. 3. Bromobenzene and O-bromophenol were both found to be hepatotoxic in English sole, as indicated by the presence of hepatocellular coagulation necrosis and fatty change in the liver, altered glutathione and ascorbic acid levels in liver tissue, elevated serum aspartate aminotransferase and alkaline phosphatase activity and increased serum glucose and triglyceride levels. 4. No evidence of nephrotoxicity was found in English sole exposed to either toxicant. 5. It is concluded that bromobenzene and O-bromophenol cannot be used as model nephrotoxicants but can be used as hepatotoxicants in English sole.

The ultrastructure and histology of hepatocellular carcinomas of English sole (Parophrys vetulus) from Puget Sound, Washington.

The cellular and subcellular morphology of hepatocellular carcinomas (HCs) from feral English sole (Parophrys vetulus) living in contaminated waterways of Puget Sound was compared with normal-appearing liver from English sole inhabiting reference sites in Puget Sound. The most distinctive feature of English sole HCs was the occurrence of large dilations of RER containing densely packed microfilamentous-like material that was often arranged in complex branching arrays that nearly filled the cytoplasm of affected hepatocytes. These arrays have not previously been reported in HCs of any other fishes. Several other characteristics also distinguished HCs from reference liver, including: the apparent proliferation of Golgi complexes and mitochondria; occurrence of torus-shaped mitochondria; elongation and/or proliferation of perisinusoidal cells; and inclusion of large intraheptocellular vacuoles.

An internal X-cell pseudotumor in a black croaker (Cheilotrema saturnum).

Cells closely resembling X-cells were the primary cellular component of a large pseudotumor in the viscera of a black croaker (Cheilotrema saturnum). The fish was captured in coastal waters off San Diego, California and was maintained at the Scripps Aquarium. After 2 years in captivity the fish exhibited extreme abdominal swelling due to a visceral mass, which weighed approximately one-fifth the total body weight. The cells associated with the pseudotumor were identified as X-cells due to their great variability in size, vesicular nuclei which stained negative for DNA and their formation of a pseudotumor. This is the first report of an X-cell pseudotumor in the visceral cavity. We agree with the hypothesis that the X-cell is a protozoan, and the description here of X-cells in a visceral pseudotumor in a fish of the family Sciaenidae associates an additional type of lesion with this enigmatic parasite.

Field and laboratory studies of the etiology of liver neoplasms in marine fish from Puget Sound.

A series of field studies was conducted between 1979 and 1985 in Puget Sound, Washington State, to investigate etiological relationships between prevalences of hepatic neoplasms in bottom-dwelling marine fish species, with emphasis on English sole (Parophrys vetulus), and concentrations of toxic chemicals in sediments and affected fish. Statistically significant (p less than or equal to 0.05) correlations have been found between the prevalences of hepatic neoplasms in English sole and the following parameters: sediment concentrations of aromatic hydrocarbons, and concentrations of the metabolites of aromatic compounds in the bile of affected sole. A significant difference (p less than 0.001) was also found between the relative concentrations of aromatic free radicals in the liver microsomes of English sole with liver lesions compared to sole without liver lesions. Laboratory studies designed to evaluate the etiology of the liver neoplasms in English sole have also yielded evidence that is consistent with the view that high molecular weight aromatic hydrocarbons, e.g., benzo[a]pyrene (BaP), are hepatocarcinogens in English sole. The current status of a series of long-term (up to 18 months) exposures of English sole and rainbow trout (Salmo gairdneri) to selected fractions of Puget Sound sediment extracts, enriched with aromatic hydrocarbons and nitrogen-containing aromatic compounds, and to individual carcinogens (e.g., BaP) is discussed.

Pathologic anatomy and patterns of occurrence of hepatic neoplasms, putative preneoplastic lesions, and other idiopathic hepatic conditions in English sole (Parophrys vetulus) from Puget Sound, Washington.

A comprehensive description of the histopathologic characteristics of a spectrum of idiopathic lesions in feral English sole (Parophrys vetulus), a bottom-dwelling flatfish, from Puget Sound, Washington State, is presented. Among these lesions are unique degenerative conditions, regeneration, storage disorders, foci of hepatocellular alteration (putative preneoplastic lesions), hepatocellular and biliary neoplasms, and nonneoplastic proliferative conditions, all of which morphologically resemble the lesions induced by various hepatocarcinogens-hepatotoxins in experimental exposures of fish and/or rodents. Results from a statistical analysis of the patterns of co-occurrence of these lesions in English sole are consistent with the concept, developed from experimental studies of liver carcinogenesis in rodents, that there are morphologically identifiable steps representing progression toward hepatic neoplasms. This is the first study in which it has been possible to demonstrate a close morphological congruity between a set of idiopathic hepatic lesions in any feral population and an established series of hepatic lesions inducible in rodents by certain hepatocarcinogens under laboratory conditions. Since sediments from the habitats occupied by the fish in this study have been shown to contain multiple hepatocarcinogens, the findings strengthen cumulative evidence that English sole are useful as indicators of exposure to hepatocarcinogens in aquatic environments.