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PubMed2004

Cloning and developmental expression of the cytochrome P450 aromatase gene (CYP19) in the European eel (Anguilla anguilla).

To characterize the involvement of the aromatase gene during the process of sex determination in the European eel (Anguilla anguilla), the expression of its gonadal form was determined during various developmental stages. The cloned cDNA from the European eel gonad (EeCYP19) contains an open reading frame of 1539 bp, encoding a deduced protein of 513 residues. The predicted amino acid sequence shows 97% identity with that of the Japanese eel, and 59-69% of identity with those of the CYP19 gonadal and brain forms of other teleost fish. Two potential initiation sites (ATG) were found downstream of the first ATG codon. A fluorescent-based method of real-time PCR was developed to quantify EeCYP19 expression. The expression levels of EeCYP19 in the gonads of adult males were approximately 12- and 30-fold lower than the levels in adult females and juvenile eels previously treated with E2, respectively. Expression of aromatase was found only in a single specimen in the control group. In contrast, no difference was found among sexes in the aromatase expression in the brain. Treatment with aromatase inhibitor (AI) of juvenile eel resulted in the total loss of aromatase expression in the gonads and brains. The results of this work revealed that AI treatment not only reduces the synthesis of estradiol, but reduces the expression levels of EeCYP19 as well. No evidence for the presence of a distinct extra-gonadal (brain) form of aromatase in the European eel could be provided.

Amino Acid Sequence↗
PubMed2004

Influence of vasostatins, the chromogranin A-derived peptides, on the working heart of the eel (Anguilla anguilla): negative inotropy and mechanism of action.

We have studied the effects of exogenous human recombinant Vasostatin-1 (VS-1), Vasostatin-2 (VS-2) and the human Chromogranin A (CGA) 7-57 synthetic peptides on the mechanical performance of the isolated and perfused working eel (Anguilla anguilla) heart. Under basal conditions, the three peptides decreased stroke volume (SV) and stroke work (SW), thus exerting negative inotropism. The VS-1-mediated negative inotropism was abolished by exposure to inhibitors of either Gi/o protein (pertussis toxin; PTx) or M1 muscarinic receptors (Pirenzepine) or calcium (Lantanum and Diltiazem) and potassium (Ba2+, 4-aminopyridine, tetraethylammonium, glibenclamide) channels, while it required an intact endocardial endothelium (EE). Using NG-monomethyl-L-arginine (L-NMMA) as an inhibitor of nitric oxide (NO) synthase (NOS), and hemoglobin as a NO scavenger, we demonstrated the obligatory role of NO signaling in mediating the vasostatin response. Pretreatment with either a specific inhibitor of soluble guanylate cyclase (GC) 1H-(1,2,4)oxadiazolo-(4,3-a)quinoxalin-1-one (ODQ), or the inhibitor of the cGMP-activated protein kinase (PKG) KT5823, abolished the VS-1-mediated inotropism, indicating the cGMP-PKG component as a crucial target of NO signaling. Of note, VS-1 was effective in counteracting the adrenergic (Isoproterenol and Phenylephrine)-mediated positive inotropism. These findings provide the first evidence that vasostatins exert cardiotropic action in fish, thus suggesting their long evolutionary history as well as their species-specific mechanisms of action.

Adrenergic Agents↗
PubMed2005

Endocrine control of Anguilla anguilla glass eel dispersal: effect of thyroid hormones on locomotor activity and rheotactic behavior.

Dispersal, one of the most important processes in population ecology, is an issue linking physiological and behavioral features. However, the endocrine control of animal dispersal remains poorly understood. Here, we tested whether and how thyroid hormones may influence dispersal in glass eels of Anguilla anguilla, by testing their influence on locomotor activity and rheotactic behavior. Glass eels were caught during their estuarine migration and treated by immersion in either a l-thyroxine (T(4)) or a thiourea (TU) solution. As measured by radioimmunoassay, T(4) and TU treatments induced, respectively, increased and decreased whole-body thyroid hormone levels relative to untreated controls. We tested a total of 960 glass eels distributed into control, and T(4) and TU treatment groups, on their swimming behavior in experimental flume tanks equipped with upstream and downstream traps that allowed us to concurrently measure both the locomotor activity and the rheotactic behavior. Compared to controls, locomotor activity significantly increased among the hyperthyroid, T(4)-treated eels, but significantly decreased among the hypothyroid, TU-treated eels. The results on rheotactic behavior suggested a more complex regulatory mechanism, since TU but not T(4) treatment significantly affected rheotactic behavior. The influence of thyroid hormones on locomotor activity suggests a central role for these hormones in the regulation of mechanisms leading to the colonization of continental habitats by glass eels. Thyroid hormones are also implicated in the control of locomotor activity in mammals and migratory behavior in birds, suggesting that these hormones represent conserved, proximate mediators of dispersal in vertebrates.

Anguilla↗
PubMed2003

Immunocytochemical studies on the pituitary gland of Anguilla anguilla L., in relation to early growth stages and diet-induced sex differentiation.

Mammalian and teleost antisera against pituitary hormones were used to identify and localize pituitary cell types in the European eel (Anguilla anguilla L.). The investigation was conducted on unpigmented glass eels of 5.6-6.2 of total body length (L(T)) caught in river mouths, then on yellow eels reared from the pigmented glass eel (or elver) stage up to 12-14 cm of L(T), in an eel farm in warm freshwater. Treated elvers were fed with commercial paste food supplemented with mature carp ovaries, containing oestradiol, that induced an early ovarian differentiation and a higher growth rate. The antisera detected seven types of immunoreactive (ir) cells, six of which were already found in glass eel adenohypophysis, suggesting differentiation of these cell types during the leptocephalus stage. In 12-14 cm treated yellow eels with small ovaries, a seventh type (ir-GtH) was detected in the proximal pars distalis; in the same animals the ir-TSH cells increased in number and size. From unpigmented glass eels to 12-14 cm yellow eels, the whole pituitary volume of controls increased nearly four times, while that of treated ones increased nearly six times. The larger volume of pituitary in treated eels was mainly due to volume increase of proximal pars distalis and rostral pars distalis. The %GH, that is the potential index of GH production, was significantly higher in treated yellow eels with gonads differentiating into ovaries than in controls; no difference was detected in %PRL between treated and control eels. The above results strongly suggest that in eels the feminizing effects of oestrogen is first exerted on the pituitary, probably through the hypothalamus, and later on the gonads.

Anguilla↗
PubMed2001

Glutathione-dependent resistance of the European eel Anguilla anguilla to the herbicide molinate.

Eels of species Anguilla anguilla were exposed to 5/4 LC50 (41.8 mg/l) of the herbicide molinate for 96 h in a time to death (TTD) test. Glutathione content (GSx, GSH, GSSG), glutathione reductase (GR) and gamma-glutamyl transpeptidase (gamma-GT) activities were determined in the liver and muscle tissues of dead and surviving (intoxicated) animals and compared to control values (non-exposed eels). TTD was positively correlated to hepatic GSH, GSH:GSSG ratio, hepatic and muscular GR, but negatively correlated to muscular GSH, which was severely depleted. Furthermore, glutathione and enzyme activities were intercorrelated, especially GSH and GR. These results indicate that eels which were able to induce GR activity, increase GSH and maintain the GSH:GSSG ratio in the liver showed an extended survival under the oxidative stress generated by molinate than those that lost glutathione homeostasis.

Anguilla↗
PubMed2003

Bile metabolites of polycyclic aromatic hydrocarbons (PAHs) in European eels Anguilla anguilla from United Kingdom estuaries.

A total of 94 European eels (Anguilla anguilla) were collected from five estuaries in the UK. The deconjugated metabolites of polycyclic aromatic hydrocarbons (PAHs) in the bile of the eels were separated using HPLC. Six PAH metabolites were identified: 1-hydroxy (1-OH) metabolites of phenanthrene, pyrene and chrysene; and the 1-OH, 3-OH and 7,8 dihydrodiol metabolites of benzo[a]pyrene (BaP). The mean concentration of the six metabolites was greatest in eels from the Tyne (49 microM) followed by the Wear (33 microM), Tees (19 microM), Thames (4 microM) and Severn (2 microM) estuaries. Although 1-OH pyrene was always the dominant compound, there were significant differences (P<0.05) between sites and between estuaries for some metabolites. Normalising the molar concentration of the bile metabolites to the bile biliverdin absorbance reduced sample variation. When the metabolites identified were each expressed as a percentage of the total detected, the metabolite profile was characteristic for each estuary.

Anguilla↗
PubMedDate not supplied

Induction of DNA strand breakage and apoptosis in the eel Anguilla anguilla.

The ability of benzo[a]pyrene, Aroclor 1254, 2-3-7-8-tetrachlorodibenzo-p-dioxin and beta-naphthoflavone to induce DNA strand breaks (SB) and apoptosis in erythrocytes of the European eel (Anguilla anguilla) was investigated following by in vivo exposure. DNA damage was evaluated by the Comet assay, while the diffusion assay was used to investigate the induction of apoptosis 7 days after a single intraperitoneal administration. 2-3-7-8-Tetrachlorodibenzo-p-dioxin induced the highest genotoxic effect, followed by benzo[a]pyrene, while the other two substances had limited effects. A significant induction of apoptosis was observed at the highest doses after exposure to benzo[a]pyrene, when DNA damage was also elevated. The occurrence of apoptotic cells after exposure to Aroclor, 2-3-7-8-tetrachlorodibenzo-p-dioxin and beta-naphthoflavone was quite variable and did not show clear dose-related responses. The role of oxidative stress in mediating DNA damage was also discussed.

Anguilla↗
PubMed2002

Biotransformation, genotoxic, and histopathological effects of environmental contaminants in European eel (Anguilla anguilla L.).

A prolonged toxicity study was carried out in young European eel (Anguilla anguilla L.) to evaluate the effects of environmental contaminants, namely, two individual standard compounds, benzo[a]pyrene (BaP) and dehydroabietic acid (DHAA), and a complex mixture, bleached kraft pulp mill effluent (BKPME). Fish were exposed to BaP (0.22, 0.45, and 0.9 microM) and BKPME (3.12%, 6.25%, and 12.5% (v/v)) for 3, 7, and 30 days and to DHAA (0.07, 0.15, and 0.30 microM) for 3, 7, 30, 90, and 180 days. The biomarkers include biotransformation and genotoxicity indicators, such as total ethoxyresorufin O-deethylase (EROD) activity and frequency of erythrocytic nuclear abnormalities (ENAs), respectively. Hematological dynamics was assessed as frequency of immature erythrocytes (IEs). Histopathological examinations were carried out for the highest concentrations and for 30 days and longer exposures. Total EROD increases significantly only after 180 days of DHAA exposure. However, significant ENA induction was generally observed during exposure to all contaminants tested. Nevertheless, some of the ENA results suggest an altered genotoxic response, which may arise either from short-term exposures to the highest contaminant levels or long-term exposures to the lowest contaminant levels. IE frequency decreased significantly after 30 days of exposure to 0.45 microM BaP and 180 days of exposure to the entire DHAA concentration range. Increased density of pigmented macrophage aggregates in 30-day BaP- and BKPME-exposed fish as well as in 90- and 180-day DHAA-exposed fish confirmed histopathological liver alterations. Bile accumulation in hepatocytes after BaP treatment, cytoplasmic vacuolization and cell atrophy following DHAA exposure, as well as liver loss of parenchymal cells in BKPME-exposed fish, were also detected. Dispersed necrosis and focal inflammation were observed in the livers of all treated groups. Fish exposed to DHAA and BKPME showed skin and gill disruption as well as kidney Malpighian corpuscle alterations. All 30-day-treated groups revealed intense spleen hemosiderosis, indicating increased erythrophagia. This splenic effect may be strongly correlated with the observed disappearance of ENAs. Neoplastic lesions were not found. A multibiomarker strategy, which includes EROD, ENA, and IE assays as well as histopathological studies, contributed to a better understanding of the global toxic process.

Abietanes↗
PubMed2003

Anguilla anguilla L. liver ethoxyresorufin O-deethylation, glutathione S-transferase, erythrocytic nuclear abnormalities, and endocrine responses to naphthalene and beta-naphthoflavone.

The effects of naphthalene (NAP) and beta-naphthoflavone (BNF) on phase I biotransformation and genotoxicity in Anguilla anguilla L. were evaluated. Phase II biotransformation and cortisol levels were also assessed in NAP-treated fish. Two groups of eels were exposed to either a NAP or a BNF concentration range (0.1-2.7 microM) for different exposure periods (2-72 h). An early significant ethoxyresorufin O-deethylation (EROD) activity inhibition was observed, especially for the highest NAP concentrations at 2-6 h exposure and for BNF at 2h exposure. However, a significant EROD activity increase was detected from 16 to 72 h exposure for NAP and from 4 to 72 h exposure for BNF. The cytochrome P450 (P450) content was not dose related. However, with regard to BNF exposure, P450 was the first biomarker to respond. Liver alanine transaminase (ALT) activity was measured as an indicator of hepatic health condition. ALT results demonstrated that the EROD activity decrease, previously described for NAP, was not related to tissue damage. Nevertheless, the highest BNF concentrations were demonstrated to induce liver damage and to impair the EROD activity response. An increased genotoxic response, measured as erythrocytic nuclear abnormalities (ENA), was observed during the first 8h NAP exposure. However, for exposures longer than 8 h, ENA frequency returned to the control levels. This response profile may reflect a considerable DNA repair capacity and/or a metabolic adaptation providing an efficient NAP biotransformation and consequent detoxification. BNF revealed no ENA alterations for all concentrations and exposure lengths. In the NAP experiment a causal relationship between immature erythrocytes (IE) and ENA frequency disappearance was not found. BNF results with regard to IE frequency revealed an ability to alter the balance between erythropoiesis and removal of erythrocytes. Liver glutathione S-transferase activity was significantly induced after 2 and 48 h NAP exposure. A cortisol-impaired response seems to occur from 4 to 24 h NAP exposure, demonstrating an endocrine disruption. However, an adaptation process seems to occur after 48 h, since the plasma cortisol had a tendency to increase. The present findings confirm the usefulness of the adopted biomarkers. The ecological risk associated with aquatic contamination by NAP was also confirmed by the present data.

Anguilla↗
PubMed2004

Enzymatic and nonenzymatic antioxidants as an adaptation to phagocyte-induced damage in Anguilla anguilla L. following in situ harbor water exposure.

Anguilla anguilla L. were caged for 8 and 48 h in harbor water of Aveiro Lagoon, Portugal. Respiratory burst activity (RBA) of peritoneal, head kidney, and gill phagocytes was measured. Lipid peroxidation (LPO) was estimated in gill, kidney, and liver. Liver ethoxyresorufin-O-deethylase (EROD) activity, cytochrome P450 (Cyt P450) content, and bile metabolites were assayed. Various antioxidant enzymes, viz., glutathione peroxidase, catalase, and glutathione S-transferase and nonenzymatic antioxidant, viz., total reduced glutathione were also studied. Harbor water xenobiotics induced a significant RBA increase in gill after 8 h; whereas in peritoneum and head kidney it increased after 48 h exposure. These responses were adversely associated with tissue-specific peroxidative damage since significant LPO increase was observed in gill (8 and 48 h), kidney (48 h), and liver (48 h). The tissue most affected was gill. Moreover, liver EROD activity, Cyt P450 content and bile metabolites remain unaltered after 8 h; in contrast, 48 h exposure showed significant EROD activity decrease and pyrene-type bile metabolites increase. Decreased EROD activity may be associated with concomitant liver damage, as increased LPO was observed after 48 h. Furthermore, the tissue-specific damage corresponded to the differences in the antioxidant potentials of the tissues, since the initial exposure period caused a significant increase in liver antioxidant activities, whereas gill and kidney showed a significant decrease, demonstrating that liver is highly adaptive to oxidative damage. However, at 48 h exposure gill, kidney, and liver showed a suppressive antioxidant effect, probably due to PAHs, since a significant induction at PAH-type bile metabolites has been seen. Our results demonstrate that phagocyte activation and associated peroxidative damage are concomitantly corroborated with enzymatic and nonenzymatic antioxidant activity differences. In addition, hepatic antioxidant induction after short-term exposure may serve as a potent biomarker for water pollutants in fish.

Anguilla↗
PubMed2004

Genotoxic and biochemical responses in caged eel (Anguilla anguilla L.) after short-term exposure to harbour waters.

European eel (Anguilla anguilla L.) were caged and exposed in situ for 8 and 48 h to the Aveiro offward fishing harbour water (HW) and to clean seawater under laboratory conditions (Control). Eel liver biotransformation (Phase I) was measured as ethoxyresorufin-O-deethylase (EROD) activity, cytochrome P450 (P450) and glutathione S-transferase (GST) activity (Phase II). Genotoxic responses were determined as blood, liver and kidney DNA strand breaks as well as erythrocytic nuclear abnormalities (ENAs). HW failed to significant increase liver EROD, GST activities and ENA frequency. Nevertheless, P450 content was significantly increased after 8 and 48 h exposure. Genotoxicity measured as DNA integrity decrease was found in blood after 8 and 48 h exposure to HW, whereas in liver and kidney, it was observed after 48 h exposure to HW. Blood, kidney and liver genotoxicity may be due to the presence of polycyclic aromatic hydrocarbons (PAHs) which are genotoxic compounds and the main HW organic contaminants.

Anguilla↗
PubMed2004

Anguilla anguilla L. plasma cortisol, lactate and glucose responses to abietic acid, dehydroabietic acid and retene.

Anguilla anguilla L. were exposed to 0, 0.1, 0.3, 0.9 and 2.7 microM abietic (AA), dehydroabietic (DHAA) acids and retene (Re) during 8, 16, 24 and 72 h. The eels plasma cortisol, glucose and lactate were measured. A significant decrease in plasma cortisol was observed at 72 h exposure to 0.9 and 2.7 microM Re. DHAA (0.1 microM) significantly decreased plasma cortisol in eels after 8 and 24 h exposure. However, a significant plasma cortisol increase was found after 16 h, 2.7 microM AA exposure and after 24 h exposure to 0.1 microM and 2.7 microM AA. Furthermore, 72 h exposure to 0.9 microM AA also induced a plasma cortisol increase. A general rise in plasma glucose was detected after all exposure periods to Re. The plasma lactate also increased after 72 h exposure to 2.7 microM AA and after 8 h exposure to 0.1 microM DHAA.

Abietanes↗
PubMed2002

Kinetics of radiolabelled silver uptake and depuration in the gills of rainbow trout (Oncorhynchus mykiss) and European eel (Anguilla anguilla): the influence of silver speciation.

We examined the influence of speciation on the kinetics of silver uptake and depuration in the gills of two freshwater fish, the rainbow trout (Oncorhynchus mykiss) which has high branchial Na(+) and Cl(-) uptake rates and is relatively sensitive to silver, and the European eel (Anguilla anguilla, yellow stage) which has low ion uptake rates and is relatively resistant to silver. Fish previously acclimated to the appropriate chloride level were exposed to 110mAgNO(3) (1.3 microg l(-1), sublethal) for 24 h in synthetic softwater with either low (10 microM) or high (1200 microM) chloride concentration, and then followed over a subsequent 67-day post-exposure period in silver-free water of the same chloride content. The exposures were therefore mainly to the free ion, Ag(+) in the low chloride water versus mainly to the neutral aqueous complex, AgCl(aq) in the high chloride water. In trout, but not in eel, water chloride is known to protect against physiological disturbances and toxicity caused by Ag(+). In both fish species, at both chloride levels, silver uptake exhibited complex kinetics. Gill silver loading occurred slowly until 6 h, then rose greatly to a peak at 12 h, followed by significant net depuration thereafter during continued exposure. By 24 h, net gill loading was three- to fivefold greater from AgCl(aq) than from Ag(+) exposure in both species, and threefold greater in trout than in eel under both conditions, with trout holding a lower fraction of the whole body burden in their gills. During the post-exposure period, depuration of silver from the gills occurred rapidly in trout, but very slowly in eel, such that gill silver burdens were greater in eel throughout the 67-day period on both an absolute and relative basis (e.g. 35% of whole body burden in eel versus <3% in trout at day 8). The kinetics of depuration were described by two phase exponential models, with break points between the fast and slow phases at 1 and 15 days for trout and eel, respectively. We conclude that speciation affects not only uptake rates but also the kinetics of depuration. When silver is loaded from AgCl(aq) it is clearly more labile than from Ag(+) exposures, with 1.6-1.8-fold greater loss rates during the fast phases in both species. Differences in branchial silver uptake between eel and trout correlate well with differences in acute toxicity, but are not as large as differences in ion uptake rates. The complex time-dependent patterns of gill loading, and the higher loading from AgCl(aq) than from Ag(+), mean that gill total silver burden is not an appropriate endpoint for biotic ligand modelling.

Adaptation, Physiological↗
PubMed2003

Internal redistribution of radiolabelled silver among tissues of rainbow trout (Oncorhynchus mykiss) and European eel (Anguilla anguilla): the influence of silver speciation.

Influence of water Ag(I) speciation on pharmacokinetics of Ag(I) during a post-exposure period was investigated in rainbow trout (Oncorhynchus mykiss) and European eel (Anguilla anguilla). The rainbow trout is sensitive to waterborne ionic Ag(+) whereas the eel is tolerant. The fish were acclimated to either of two chloride concentrations, 10 or 1200 microM, in synthetic soft water and then exposed to a sublethal 24-h pulse of 1.3 microgl(-1) of 110mAg(I) added as 110mAgNO(3) in these waters. The protocol provided exposures to mainly the free ion Ag(+) (low chloride water) or mainly AgCl(aq) (high chloride water). Contents and concentrations of 110mAg(I) in tissues and body fluids were then monitored over a 67-day post-exposure period in Ag(I)-free water of the same chloride levels. Changing the speciation of Ag(I) in the water had no effect on the whole body load of 110mAg(I), but did result in differences in internal distribution. In trout, changing water Ag(I) speciation significantly altered elimination or accumulation of Ag(I) in several body compartments. Notably, trout exposed to AgCl(aq) eliminated 110mAg(I) from the kidney more quickly than trout exposed to Ag(I) primarily as Ag(+). This elimination was matched by higher concentrations of 110mAg in liver of trout exposed to Ag(I) primarily as AgCl(aq). In eel, shifting speciation from Ag(+) to AgCl(aq) hastened elimination of 110mAg(I) from mid and posterior intestine and increased 110mAg(I) retention in kidney. While there was no difference between the two fish species in whole body 110mAg(I) load, most internal body compartments of trout had higher 110mAg(I) concentrations than those in eel early in the experiment. Because tissue-specific elimination times were longer in eel than in trout, these differences were generally cancelled by the end of the 67-day depuration period. The only exception was the liver, which in trout continued to accumulate 110mAg(I) throughout the experiment but in eel remained unchanged. The combined effect of 110mAg(I) movements in the two species was that trout retained all their accumulated 110mAg(I) through the 67-day period, whereas the body burden of 110mAg(I) in eel was reduced to half initial values by day 67.

Adaptation, Physiological↗
PubMed1999

High density lipoprotein (HDL), and not albumin, is the major palmitate binding protein in New Zealand long-finned (Anguilla dieffenbachii) and short-finned eel (Anguilla australis schmidtii) plasma.

Plasma from two members of the teleost Anguillidae family, the New Zealand long-finned (Anguilla dieffenbachii) and short-finned eels (Anguilla australis schmidtii), were examined. Agarose gel electrophoresis showed both species had a major anionic diffuse protein band migrating at approximately the same position as human albumin, and autoradiography showed this protein bound [14C]palmitic acid, but not 63Ni2+. Cellulose acetate electrophoresis followed by Oil Red O staining suggested that this band was a lipoprotein. Two-dimensional electrophoresis of plasma showed the absence of a significant albumin band at approx. 65 kDa, and that the palmitate binding band appeared to be composed of at least three proteins, with the major protein running at 30 kDa. N-Terminal sequencing of the palmitate binding band indicated major sequences of DAPAPP(S)QLED- for long-finned eel and DAPAPPSQLEHV- for short-finned eel, confirming their identities as apo-AI, the major apolipoprotein of high density lipoprotein (HDL). When ultracentrifugation was used to separate the lipoproteins of each species, the anionic palmitate binding protein was found solely in the lipoprotein fractions. There was no evidence of albumin in plasma from either eel, and it appears that in its absence HDL takes on the role of fatty acid transport.

Anguilla↗
PubMed1998

Cu uptake, metabolism and elimination in fed and starved European eels (Anguilla anguilla) during adaptation to water-borne Cu exposure.

64Cu accumulation and total Cu concentrations were measured in gill filaments, plasma, liver and bile of fed and starved European eels (Anguilla anguilla) during 28 days of exposure to 12 and 94 micrograms Cu l-1. Branchial Cu uptake was found to be 0.03 and 0.13 microgram Cu g gill filament-1 h-1 at 12 and 94 micrograms Cu l-1, respectively, throughout 28 days of Cu exposure. At least during exposure to 94 micrograms Cu l-1, the basolateral membrane seemed to be the rate limiting step for branchial Cu uptake. The plasma accounted for at least 70% of the 64Cu labelled Cu present in the blood at both Cu concentrations. The plasma Cu concentration was gradually increased from 0.64 to 1.92 and 2.68 micrograms ml-1 during the first 6 days of Cu exposure; it was partly normalized to 1.07 and 1.45 micrograms ml-1 after 28 days of exposure to 12 and 94 micrograms Cu l-1, respectively, despite a constant branchial Cu uptake, indicating an increased turnover of plasma Cu. The hepatic accumulation of 64Cu was similar in fed and starved eels (0.55 microgram Cu g liver-1 h-1) at both Cu concentrations during the 28 days of exposure. The hepatic total Cu concentrations were lower in the fed fish compared to the starved fish and increased only marginally from 3 to 28 days of exposure indicating a high turnover of hepatic Cu during acclimation. Hepatic Cu elimination, measured as 64Cu in the bile, was highest in the fed fish at the highest Cu concentration and was stimulated during acclimation at least in the starved fish.

Adaptation, Physiological↗
PubMed2000

Catalytic properties of CYP1A isoforms in the liver of an agnathan (Lampetra fluviatilis) and two species of teleost (Pleuronectes flesus, Anguilla anguilla).

The catalytic activity of CYP1A isoforms and the effect of mammalian CYP1A-specific inhibitors in liver S9 fractions were studied in an agnathan (River lamprey, Lampetra fluviatilis, 30-33 cm) and in two species of teleost fish (European flounder, Pleuronectes flesus, 11-18 cm and common eel, Anguilla anguilla, 31-48 cm). Ethoxyresorufin O-deethylation (EROD), caffeine N-demethylation/C-oxidation and phenacetin O-deethylation (POD) activity increased 3-4-fold in flounders and 17-46-fold in eels, 5 days after fish were injected (i.p.) with 100 mg kg(-1) benzo(a)pyrene (B[a]P). In lampreys, basal EROD activity was very low and no increase in activity was observed following exposure to B[a]P. While the apparent Michaelis constant (K(m)) for each assay showed only small changes after B[a]P injection, maximum reaction velocity (V(max)) values increased by up to 19- and 84-fold for EROD activity, 4- and 35-fold for caffeine-related metabolism and 4- and 19-fold for POD activity in flounders and eels, respectively. The mammalian CYP1A2 inhibitor furafylline (50 microM-1 mM) reduced activity in the EROD, caffeine and POD assays to 65, 21 and 20% of control values in flounders and to 85, 10 and 5% of control values in eels, respectively. By contrast, low concentrations (0.025-0.050 microM) of the mammalian CYP1A1 inhibitor ellipticine completely abolished EROD activity, but had no effect (up to 1 mM) on caffeine metabolism or POD activity in either species. While the inhibitor studies strongly suggest that two separate enzymes are present in flounders and eels, the monophasic Michaelis-Menten kinetics obtained in all the assays imply that only a single CYP1A protein is present that has substrate and inhibitor specificities characteristic of both mammalian CYP1A1 and CYP1A2 isoforms.

Anguilla↗
PubMed1997

Composition and structure of helminth communities in eels Anguilla anguilla from Italian coastal lagoons.

The composition and diversity of total and intestinal helminth component and infracommunities were determined in eels Anguilla anguilla from four shallow coastal lagoons near Rome. The lagoons differed principally in respect of their salinity. Only Lake Burano still received an input of freshwater, and both freshwater and marine helminth species were found in eels. In the other three lagoons all helminth species were marine or euryhaline and communities were dominated by digeneans. At component community level these three lagoons were far more similar to each other than to Lake Burano, but this distinction almost disappeared at infracommunity level. Species richness and diversity declined with increasing salinity at component community level but not at infracommunity level. Despite the changes and differences in helminth community composition, intestinal helminth infracommunity structure was very similar in all four lagoons and to that in eels from freshwater localities in Britain. The findings thus provide further and unexpected support for the view that some factor(s) other than supply side ones must be operating to produce the fundamental structural similarity in helminth communities in eels.

Anguilla↗