Parasitology of the English Sole, Parophrys vetulus Girard in Oregon, U.S.A.

A total of 877 juvenile English sole ( Parophrys vetulus Girard) from the Yaquina Bay estuary and742 juvenileandadultsole from the Pacific Ocean off Oregon were examined forparasites. Fifteen species of parasites were found in juvenile English sole on the estuarine nursery ground. Differences in the prevalence and intensity of parasite infection between size classes of juvenile sole and between sole occupying the upper and lower estuary were determined. An additional 14 parasite species were found in offshore English sole, bringing the total observed in all fish to 29 species. Parasites acquired only in the estuary included the microsporidan Glugea stephani , the acanthocephalan Echinorhynchus lageniformis , and the nematode Philometra americana . Those acquired only in offshore areas included the trematodes Otodistomum veliporum and Zoogonus dextrocirrus , the leech Oceanobdella sp. and three species of copepods. An attempt to use parasite data to indicate the presence of distinct English sole stocks along the Oregon coast was inconclusive.     

A new species of marine leech (Hirudinea: Piscicolidae) from the north-eastern Pacific Ocean, parasitic on the English sole, Parophrys vetulus Girard

The marine piscicolid leech Calliobdella knightjonesi sp. nov. from the Oregon coast is described and illustrated. Mature individuals do not exceed 10 mm in length, have faint reddish-brown segmental pigment bands, small pulsatile vesicles, six pairs of testisacs and a well-developed seminal receptacle connected to the posterior portion of the bursa by paired tissue strands. The leech lacks eyes and ocelli.     

Epizootic characteristics of hepatic and renal lesions in English sole, Parophrys vetulus, from Puget Sound

English sole, Parophrys vetulus , from eight locations within Puget Sound, U.S.A. were examined for histological abnormalities of the liver and kidney. A total of 1083 fish were collected between February 1979 and June 1984 in nine separate sampling periods. Idiopathic lesions, i.e. lesions of unknown cause, were classified into eight hepatic and four renal lesion categories. The significance of certain potential risk factors associated with lesion prevalences, including age, gender, site of capture, season of capture and year of capture, were assessed by logistic regression. The odds ratios for the significant risk factors were also calculated. Site of capture and age of the examined fish were found to be the most important factors for most of the observed lesion types, e.g., hepatic neoplasms and foci of cellular alteration, whereas year and season of capture were significant influences for only a few lesion types, e.g., specific and nonspecific degenerative/necrotic lesions of the liver. Gender of examined fish was not found to be a significant factor for any of the lesion types. These results emphasize the importance of including certain potential influences, such as age and season of capture, in the analysis of field-collected organ pathology data.     

Trypanosoma pacifica sp. n. from the English Sole Parophrys vetulus Girard from Oregon*

Trypanosoma pacifica sp. n. is described from the blood of the English sole, Parophrys vetulus Girard, from Oregon. The total length averages 35.9 μm of which 15.4 μm is free flagellum. Comparisons are made with other trypanosomes reported from related species of fishes and with those reported from marine fishes adjacent to North America.     

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.     

Oxidative DNA damage in tissues of English sole (Parophrys vetulus) exposed to nitrofurantoin

Juvenile English sole were exposed intramuscularly to nitrofurantoin (NF) and the levels of 8-hydroxy-2′deoxyguanosine (8-OH-dG) in liver, kidney and blood were determined using reversed-phase HPLC with electrochemical detection. Identification and quantitation of the 8-OH-dG in the samples was accomplished by comparison with standard 8-OH-dG, which was characterized by UV spectroscopy and fast-atom bombardment mass spectrometry. The levels of hepatic 8-OH-dG increased (r² = 0.59, P = 0.015) with the dose of NF (0.10 – 10 mg NF/kg fish). In kidney and blood, however, the levels of 8-OH-dG were significantly higher than controls only at the highest dose tested. The level of binding in liver ranged from 0.37 to 0.76 fmol 8-OH-dG/μg DNA. The levels of hepatic 8-OH-dG reached a maximum (approx. 1 fmol 8-OH-dG/μg DNA) between 1 and 3 days after exposure, followed by a decrease to control levels (approx. 0.25 fmol 8-OH-dG/μg DNA) at 5 days post-exposure. These data demonstrate the first direct evidence for the formation of oxidized DNA bases resulting from the metabolism of a nitroaromatic compound by fish.     

Effects of low temperature on the development of the microsporidan Glugea stephani in English sole (Parophrys vetulus)

Glugea stephani requires temperatures above 15 C for development in juvenile pleuronectid flatfishes in Yaquina Bay, Oregon. The effect of low temperature (10 C) on the development of recently established parasites was tested experimentally in juvenile English sole (Parophrys vetulus). Low temperature arrested parasite development, but did not kill the protozoan which resumed development on return to 19-20 C after as long as 42 days at 10 C. No parasites detectable with the light microscope were found in fish examined after 70 days at 10 C. Although most juvenile English sole move permanently from the estuary to cooler ocean waters in fall and do not contribute to the continuation of the parasite life cycle, the cycle may be maintained by low numbers of English sole that overwinter in the estuary.     

Hepatotoxic effects of CCl4 on English sole (Parophrys vetulus): possible indicators of liver dysfunction

Selected serum parameters (enzyme activities and triglycerides) and liver glutathione and vitamin C concentrations were measured in English sole (Parophrys vetulus) after i.p. injection of carbon tetrachloride (CCl4), a hepatotoxin in fish. Serum lactate dehydrogenase (LDH), alkaline phosphatase (AP) and glutamate dehydrogenase (GDH) activities and the concentration of triglycerides increased in a dose-dependent manner 24 hr post injection. Concentrations of glutathione (reduced and oxidized) and ascorbic acid (vitamin C) in liver did not change in response to CCl4 toxicity 24 hr post injection. These studies indicate that serum AP activity and triglyceride concentrations can be useful in assessing the effects of CCl4-induced liver toxicity in this species of marine fish. Serum LDH and GDH activity should be used with some caution in assessing liver damage in English sole, as other tissues represent more likely sources for serum activity. The levels of liver antioxidants do not appear to be significantly affected, 24 hr post injection, by this particular hepatotoxin.     

Multiplication of Trypanosoma pacifica (Euglenozoa: Kinetoplastea) in English sole, Parophrys vetulus, from Oregon coastal waters

Multiplication of Trypanosoma pacifica was common in the fish host from observations of live flagellates and Giemsa-stained blood smears. Multiplication began with the elongation of the kinetoplast, thickening of the posterior portion of the body, and appearance of a new flagellum near the kinetoplast. The new flagellum was very rigid when less than 3 microm in length, but it became flexible as it elongated. When the new flagellum was approximately 12 microm in length, cell division began and the kinetoplast also began to divide. The timing of nuclear division was variable. Generally, it did not occur until division of the kinetoplast had begun, but occasionally binucleate individuals were observed before cell or kinetoplast division was apparent. As division continued, 1 nucleus migrated past the dividing kinetoplast into the future daughter trypanosome. Finally, the kinetoplast completed division and the trypanosomes separated. Cell division was unequal, with the daughter trypanosome being smaller than the parent and with a more weakly developed undulating membrane.     

Formation and persistence of benzo[a]pyrene-diolepoxide-DNA adducts in liver of English sole (Parophrys vetulus)

The formation of DNA adducts from the carcinogenic environmental pollutant benzo[a]pyrene (BaP) was investigated in liver of English sole (Parophrys vetulus), a fish species that exhibits a high prevalence of liver neoplasms in several polycyclic aromatic hydrocarbon (PAH)-contaminated areas of Puget Sound, WA. Analysis by the 32P-postlabeling assay of hepatic DNA digests from English sole exposed parenterally to BaP showed the presence of BaP-diol epoxide (BaPDE)-DNA adducts. When English sole were injected with 2-15 mg BaP/kg body wt., one major adduct was detected and was identified as the anti-BaPDE-DNA adduct. Moreover, in English sole sampled at 1, 28 and 60 days post-exposure to 15 mg BaP/kg body wt., there was no significant change in the level of the anti-BaPDE-DNA adduct. The autoradiographs of 32P-labeled hepatic DNA digests from fish exposed to 100 mg BaP/kg body wt. showed an elongated spot suggesting the presence of more than one adduct. Chromatography on large polyethyleneimine sheets (20 x 20 cm) showed 2 spots with the same chromatographic characteristics as those of syn- and anti-BaPDE-deoxyguanosine adduct standards. Mild acid hydrolysis of hepatic DNA of English sole, exposed to 100 mg BaP/kg body wt., also revealed the presence of tetrols derived from both anti- and syn-BaPDE, thus confirming the presence of syn- and anti-BaPDE. In fish exposed to 2-100 mg BaP/kg body wt., a linear (0.996) dose response for anti-BaPDE-DNA adduct formation was observed. The results from this study offer the first direct evidence for the formation of the suspected ultimate carcinogen, BaPDE, in liver of English sole exposed to BaP in vivo and thus further support the hypothesis that exposure to PAHs is an important factor in the etiology of hepatic neoplasms in English sole from contaminated sites.     

Nucleotide sequence and tissue distribution of the human mitochondrial aspartate aminotransferase mRNA

The cDNA of human mitochondrial aspartate aminotransferase (E.C.2.6.1.1.) was isolated from a human liver cDNA library using a rat mitochondrial aspartate aminotransferase cDNA as probe. The sequence of this cDNA gives a predicted aminoacid sequence for the human presequence and for the human mature protein exhibiting respectively 93% and 95% homology with rat sequences. A Northern blot of total RNA, isolated from various human tissues and hybridized with this cDNA, revealed a single 2.4 Kb RNA band. Mitochondrial aspartate aminotransferase RNA was clearly detected in human kidney, placenta, stomach and spleen as well as in both fetal and adult liver.     

Intertidal recruitment and feeding in relation to optimal utilization of nursery areas by juvenile English sole (Parophrys vetulus: Pleuronectidae)

Synopsis Juvenile Parophrys vetulus 19–102 mm were collected on an intertidal flat in Humboldt Bay during 1976–1977. Recently metamorphosed fish that fed almost exclusively on harpacticoid copepods and other epibenthic crustaceans were found in large numbers during spring and early summer. A feeding transition occurred among fish 50–65 mm in length, and infaunal polychaetes were the dominant prey of fish greater than 65 mm. Increases in the feeding niche width and average size of prey items, as well as a decrease in the number of prey items per stomach, accompanied growth.The disappearance of English sole from intertidal areas in early fall at an average size of 82 mm and their subsequent residence in subtidal channels until a size of 140 mm suggest a size-depth segregation within the nursery ground. The advantages of such a distribution are discussed in terms of optimal foraging and reduction of intra- and interspecific competition.     

Xenobiotic metabolizing enzymes in spawning English sole (Parophrys vetulus) exposed to organic-solvent extracts of marine sediments from contaminated and reference areas

Female English sole (Parophrys vetulus) within 1-2 days of spawning were exposed by i.m. injection to organic-solvent extracts of marine sediments at the following doses: Eagle Harbor (EHSE, contaminated site)--6.8 mg aromatic hydrocarbons (AHs)/kg body wt; Duwamish Waterway (DSE, contaminated site)--0.52 mg AHs and 0.040 mg chlorinated hydrocarbons (CHs)/kg body wt; Hood Canal (HCSE, reference site)--0.00090 mg AHs/kg body wt. Hepatic aryl hydrocarbon hydroxylase (AHH) activity, measured at spawning, was induced 10-, 23-and 2-fold by EHSE, DSE and HCSE, respectively, compared to sham and vehicle controls. Hepatic glutathione-S-transferase and epoxide hydrolase activities were not affected by any treatment. EHSE, but not DSE or HCSE, inhibited spawning (P less than 0.01) in 36% of the exposed fish and hepatic AHH activity in the non-spawning fish was significantly (P less than 0.05) higher than in the fish that did spawn. These results suggest a potential for reproductive toxicity in benthic fish after exposure to sediment-associated contaminants.     

Diurnal variations of nitrite reductase, glutamine synthetase, glutamate synthase, alanine aminotransferase and aspartate aminotransferase in barley leaves

Diurnal variations of in vitro activity of 5 enzymes of nitrogen metabolism were studied. Barley ( Hordeum vulgare L. cv. Herta) seedlings were grown in 8 h short days, in daylight or under fluorescent lamps. During, the photoperiod nitrite reductase (EC 1.7.7.1) increased by an average of 18% in daylight and 10% under fluorescent lamps. Glutamine synthetase (EC 6.3.1.2) activity increased by 14 and 10%, respectively. The increase in enzyme activity reflected the overall increase in soluble proteins which was 8% in daylight and 3% under fluorescent lamps. Alanine aminotransferase (EC 2.6.1.2) increased by 82% in daylight and 37% under fluorescent lamps. Desalting of the extracts did not alter the enzyme activity and thus supported the assumption that changes in extractable enzyme activity are due to changes in the amount of (active) enzyme protein. Glutamate synthase (EC 1.4.7.1) activity did not show regular diurnal variations, and aspartate aminotransferase (EC 2.6.1.1) activity was almost constant.     

Intramitochondrial localization of alanine aminotransferase in rat-liver mitochondria: comparison with glutaminase and aspartate aminotransferase

Summary The removal of the outer mitochondrial membrane and hence of constituents of the intermembrane space in rat-liver mitochondria using digitonin showed that phosphate-dependent glutaminase, alanine and aspartate aminotransferase were localized in the mitoplasts. Further fractionation of mitoplasts following their sonication resulted in 90% of glutaminase, 98% of alanine aminotransferase and 48% of aspartate aminotransferase being recovered in the soluble fraction while the remainder of each enzyme was recovered in the sonicated vesicles fraction. These results indicated that glutaminase and alanine aminotransferase were soluble matrix enzymes, the little of each enzyme recovered in the sonicated vesicles fraction being probably due to entrapment in the vesicles. Aspartate aminotransferase had dual localization, in the inner membrane and matrix with the high specific activity in sonicated vesicles confirming its association with the membrane. Activation experiments suggested that the membrane-bound enzyme was localized on the inner side of the inner mitochondrial membrane.