Comparative studies on the chromosomes of the European eel (Anguilla anguilla L.) and the American eel (Anguilla rostrata Le Sueur).
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Development of hepatic CYP1A and blood vitellogenin in eel (Anguilla anguilla) for use as biomarkers in the Thames Estuary, UK.
The potential of eel (Anguilla anguilla) as a monitoring species for the Thames Estuary, UK, was examined. Hepatic cytochrome P4501A [7-ethoxyresorufin O-deethylase (EROD) activity] and blood vitellogenin (Western analysis) were investigated as biomarkers of exposure to, respectively, organic contaminants and to contaminants showing estrogenic activity. Hepatic microsomal EROD activities in A. anguilla from seven sites in the Thames Estuary in May 1998 varied three-fold (111 +/- 24 to 355 +/- 42 pmol min-1 mg protein-1) (mean +/- S.E.M.) and showed correlation with salinity; however, the latter relationship was not maintained at other times of the year. The range of EROD activities was two- to eight-fold higher than the 37 +/- 8 pmol min-1 mg-1 for A. anguilla from the relatively clean Tamar Estuary. beta-Naphthoflavone treatment (5 mg kg-1 wet wt.; 2 days) of Thames A. anguilla produced a two-fold increase in hepatic microsomal EROD activity. Comparing the Thames EROD data with those for A. anguilla from well-characterised contaminated sites in the Netherlands (Van der Oost, R., Goksøyr, A., Celander, M., Heida, H., & Vermeulen, N. P. E. 1996. Aquatic Toxicology, 36, 189-222), the Thames is suggested to be moderately impacted by polycyclic aromatic hydrocarbons and related contaminants. 17-beta-Estradiol treatment produced the appearance of a plasma protein of 211 Kd app. mol. wt. (recognised by antibodies to vitellogenin of Morone saxatilis), but putative vitellogenin could not be detected in A. anguilla from selected sites in the Thames Estuary.
Anguilla↗
Carbonic anhydrase activity in tissues of the icefish Chionodraco hamatus and of the red-blooded teleosts Trematomus bernacchii and Anguilla anguilla.
Carbonic anhydrase (CA) activity was measured in blood, intestine, kidney and gill of two Antarctic teleosts, the haemoglobinless Chionodraco hamatus and the red-blooded Trematomus bernacchii, and of the temperate teleost Anguilla anguilla. In all species, the highest CA activity was in the gills, with the greatest activity in C. hamatus. CA activity in the blood was highest in A. anguilla, but none was detected in the blood of C. hamatus despite the presence of plasma CA inhibitors. The enzyme was present but its activity was low in the intestine and kidney of all three species. The existence of very high CA activity in C. hamatus gills compared with the red-blooded species was investigated further by isolating and characterising the branchial cytosolic CA isoforms. The turnover rate of the C. hamatus isoform was significantly higher than that of T. bernacchii and A. anguilla. The isoforms from both the Antarctic species exhibited lower apparent K(m) (K(m,app)) and heat stability than those from A. anguilla. Sensitivity to sulphonamides was similar in all species and was within the range of the mammalian CA II isoform. The branchial CA isoforms of C. hamatus, T. bernacchii and A. anguilla displayed relative molecular masses of 28.9, 29.9 and 31.2 kDa, respectively. The results suggest that the hemoglobinless teleost possesses a different branchial cytosolic CA isoform from that of red-blooded teleosts.
Anguilla↗
Helminth communities of European eels Anguilla anguilla (Linnaeus, 1758) from the Vistula Lagoon and Puck Bay, Poland.
Within 2001-2002 a total of 621 eel Anguilla anguilla (L., 1758) (488 from the Vistula Lagoon and 133 from the Puck Bay) were examined. Fifteen parasite taxa were recovered: Pseudodactylogyrus anguillae (Yin et Sproston, 1948), Brachyphallus crenatus (Rudolphi, 1802), Deropristis inflata (Molin, 1859), Diplostomum spp., Bothriocephalus claviceps (Goeze, 1782), Proteocephalus macrocephalus (Creplin, 1825), Anguillicola crassus (Kuwahara, Niimi et Itagaki, 1974), Camallanus lacustris (Zoega, 1776), Cystidicola farionis Fischer, 1798, Hysterothylacium aduncum (Rudolphi, 1802), Raphidascaris acus (Bloch, 1779), Acanthocephalus anguillae (Müller, 1780), A. lucii (Müller, 1776), Echinorhynchus gadi Müller, 1776, and Pomphorhynchus laevis (Müller, 1776), representing Monogenea, Digenea, Cestoda, Nematoda, and Acanthocephala, respectively. Ten of these taxa occurred in the Vistula Lagoon, while fourteen were noted in the Puck Bay. P. anguillae, Diplostomum spp., C. lacustris, C. farionis and P. laevis were not found in the lagoon eels, while B. crenatus did not occur in the bay. Anguillicola crassus was the most frequently found parasite (Vistula Lagoon: prevalence 75%, mean intensity 6.9 specimens; Puck Bay: 74.4%, and 8.3 specimens, respectively). Pseudodactylogyrus anguillae was recorded for the first time in the Puck Bay.
Anguilla↗
Stegodexamene anguillae (Digenea: Lepocreadiidae), an intestinal parasite of eels (Anguilla spp.) in New Caledonia.
Adult trematodes of the genus Stegodexamene Macfarlane, 1951 were recorded from the intestines of two species of freshwater eels, the Pacific short-finned eel Anguilla obscura Günther and the speckled longfin eel Anguilla reinhardtii Steindachner from the Pouembout and La Foa Rivers, New Caledonia, South Pacific. They were identified as Stegodexamene anguillae Macfarlane, 1951, a species previously reported only from eels in New Zealand. The morphology of adult S. anguillae seems to be practically identical with that of the later described Stegodexamene callista Watson 1984 from eels in Australia. The finding of S. anguillae in A. obscura and A. reinhardtii in New Caledonia represents new geographical and host records. In addition to this trematode, two larval specimens of a cestode of the order Proteocephalidea were found in the intestine of one A. obscura from the Pouembout River. No monogenean was found in eels of New Caledonia.
Anguilla↗
Responses of European eel (Anguilla anguilla L.) in two polluted environments: in situ experiments.
European eels (Anguilla anguilla L.) were caged for 8 and 48 h in two polluted areas to assess the contamination cleanup process in the Vouga River 2 years after the official closing of a bleached kraft pulp mill effluent (BKPME; experiment 1), and to monitor the effects induced by contaminated offward fishing harbor waters (experiment 2). Plasma cortisol, glucose, and lactate were evaluated as stress responses. In experiment 1, plasma cortisol, glucose, and lactate increased after 8 h of exposure in site 3, which is located farthest from the deactivated sewage outlet. However, A. anguilla seemed to adapt after 48 h of exposure in site 3, because all three parameters returned to control levels. Plasma glucose also significantly increased after 8 h of exposure at sites 1, 2, and 3, returning to control levels after 48 h. Plasma lactate levels increased after 8 h of exposure at site 3 and after 48 h of exposure at site 1. In experiment 2, A. anguilla exposed to contaminated harbor waters increased their plasma lactate after 8 and 48 h of exposure, whereas their cortisol and glucose plasma were elevated only after 48 h of exposure. The results demonstrate that even 2 years after the official closing of the BKPME sewage outlet, the river Vouga water remains contaminated by the sediment associated chemicals. Because the fishing harbor induced in A. anguilla the same type of stress responses, it is also an area of concern. The adopted stress parameters allied to a caging strategy are recommended for future environmental monitoring assessments.
Anguilla↗
Hepatic 7-ethoxyresorufin O-deethylase activity in eel (Anguilla anguilla) from the Thames Estuary and comparisons with other United Kingdom Estuaries.
Hepatic microsomal 7-ethoxyresorufin O-deethylase (EROD) activities (indicative of exposure to polycyclic aromatic hydrocarbons (PAHs) and polychlorobiphenyls (PCBs)) were measured in eel Anguilla anguilla from the Thames Estuary. Fish were collected from up to 13 sites during November 1997, May and August 1998 and October 1999. Throughout this period no clear seasonal variation could be identified at every site along the Thames. However, during the summer months, fish sampled from sites in the middle to the upper estuary (Woolwich, Greenhithe and West Thurrock) reported up to 3-fold higher EROD activities compared to sites either at the upper reaches (Richmond and Brentford) at the same time of the year, or fish sampled in winter, along the entire length of the estuary. A laboratory exposure experiment demonstrated a 3-fold elevation of EROD activity 2 days after injection with beta-naphthoflavone (beta-NF). However, higher levels of activity could be determined in fish sampled from the Weston canal near the Merseyside. The lowest levels of A. anguilla EROD activity were observed in fish sampled from the upper reaches of the River Tamar, Devon, and were comparable to activities determined in fish from the Wear and Humber estuaries. A. anguilla sampled along the Thames, Tyne and Tees estuaries reported between 2.5- and 7-fold higher EROD activities compared to fish collected from the Tamar. These results indicate that a low to moderate induction of A. anguilla CYP1A had occurred (indicative of low to moderate exposure to PAHs and planar PCBs) in fish collected from the Thames, Tyne, Wear, Tees, Humber and Tamar estuaries. However, the highest level of EROD activity was observed in fish from the Weston Canal (Merseyside).
Anguilla↗
Seasonal variation and estradiol-dependent elevation of Thames estuary eel Anguilla anguilla plasma vitellogenin levels and comparisons with other United Kingdom estuaries.
Eel Anguilla anguilla plasma vitellogenin was investigated as a biomarker of exposure to environmental compounds with estrogenic activity, along the tidal course of the Thames Estuary, UK. A. anguilla was chosen as a sentinel species because of their wide distribution, robustness in field and laboratory studies and also because they have a characterised normal intersex' condition where the gonad contains both developing male and female gonadal cells termed a Syrski organ. Following laboratory exposure to 17beta-estradiol (intraperitoneal injection), a plasma protein (approx. 211 kDa apparent molecular weight) was detected by monoclonal antibodies to vitellogenin of striped bass (Morone saxatilis). Western and dot blot analyses were developed and vitellogenin was isolated from 17beta-estradiol-treated fish to calibrate the quantification of the blots by image analysis. The limits of sensitivity for the Western and dot blots were 100 and 10 ng vitellogenin/ml, respectively. Levels of vitellogenin in Thames estuary samples were below the detection limits of the Western but not the dot blot, and showed no statistically significant site-specific (10 sites) and seasonal-specific (May, August, November) differences. Values were observed to be low, between 11 and 165 ng/ml, compared with 17-50 mg/ml for 17beta-estradiol-treated eels. Similar low levels of plasma vitellogenin were determined in fish sampled along the Tyne, Wear, Tees or Humber estuaries, or the Weston canal Liverpool, with mean plasma vitellogenin levels varying between 44 and 82 ng/ml. These levels of vitellogenin in A. anguilla plasma were observed to be consistent with the known biology of the eel. Immature females, or fish with syrski organs, reported both lower levels and smaller variation of plasma vitellogenin concentrations whereas the highest plasma vitellogenin concentrations were determined in fish above 45 cm consistent with female fish. These results indicate inter-species variation between the plasma vitellogenin concentrations of A. anguilla and other published fish studies undertaken along the same estuaries.
Anguilla↗
Long-term investigation of the composition and richness of intestinal helminth communities in the stocked population of eel, Anguilla anguilla, in Neusiedler See, Austria.
Data from a long-term study of the intestinal helminth parasite community of eels, Anguilla anguilla, stocked into the shallow eutrophic Neusiedler See, Austria, were collected over an 8 year period (1994-2001). In total, 720 eels from 2 sampling sites were examined. The parasite community showed characteristics similar to those in the natural eel populations in rivers of the UK and mainland Europe: it was species poor, with only 5 species (Acanthocephalus lucii, Acanthocephalus anguillae, Raphidascaris acus, Proteocephalus macrocephalus, Bothriocephalus claviceps) comprising the component community and a maximum infracommunity richness of 4 species. Over the period, the intestinal parasite community of the sampling site in Illmitz, which was originally dominated by A. lucii, changed. As levels of A. anguillae increased to a point at which it dominated the community, diversity increased whilst dominance of a single species decreased. By contrast the community in the southern sampling site remained rather constant with a continuous high infection level of A. anguillae and low abundance of A. lucii. Both acanthocephalan species exhibited higher infection levels in larger eels and in different seasons of the year and the infection parameters were significantly different between the years of study. The significant differences in the infection levels of the 2 acanthocephalan species at the 2 sampling sites were surprising as both acanthocephalan species use the same intermediate host, Asellus aquaticus, and the sampling sites were in close proximity and were similar in terms of water quality, host size and invertebrate abundance. Differences in the fish communities of the 2 sampling sites and eel movements rather than interspecific competition are discussed as possible explanations for the differences in the parasite communities of the 2 sampling sites.
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Parasitological study of a population of Tiber River eels (Anguilla anguilla).
A study was conducted on the parasitofauna of Anguilla anguilla caught in the section of the Tiber river south of Rome (Italy). The sample examined consisted of 417 specimens that were classified by stage (elver, young yellow eel, yellow eel and silver eel) and season of sampling. The following species were identified: Trypanosoma granulosum, Eimeria anguillae, Myxidium giardi, Sphaerospora reichenowi, Myxobolus sp., Trichodina anguilli, Ichthyohodo sp., Gyrodactylus anguillae, Bothriocephalus claviceps, Proteocephalus macrocephalus, Raphidascaris acus, Acanthocephalus clavula. Each parasite species was considered in its prevalence according to stage and season. For some parasite the pathogenic effect was also examined in a histological study of serial sections. The results of the statistical analysis of the single parasitic infection as related to different seasons pointed to marked seasonal trends only in some species (Trypanosoma granulosum and Gyrodactylus anguillae). The intensity of infection assessed for acanthocephalan infection only was found to affect neither weight nor condition index (weight/length3). No difference in weight or condition index was found to be ascribable to the presence or absence of a species. An attempt to find a rule-of-thumb algorithm to gauge the overall effect to parasitic diseases failed to give sufficiently significant results.
Acanthocephala↗
Effects of diazinon exposure on cholinesterase activity in different tissues of European eel (Anguilla anguilla).
Cholinesterase (ChE) activity was measured in brain, plasma, and whole eye of Anguilla anguilla experimentally exposed to a sublethal concentration of 0.042 mg/liter (0.50 of the 96-hr LC50) of the organophosphorous pesticide diazinon. Whole eye was the tissue which revealed higher values of ChE activity (8.17 micromol/min/g) in nonexposed animals. Brain, plasma, and whole eye ChE activity of A. anguilla was inhibited at 6, 24, 48, 72, and 96 hr of diazinon exposure. Pesticide induced significant inhibitory effects on the ChE activity of this species ranging from >70% inhibition in brain tissue to >90% in plasma samples. Brain and plasma presented technical difficulties in their collection. Due to its ease of collection, high ChE activity levels, and sensitivity to ChE pesticide inhibition, the use of whole eye is recommended for post-mortem ChE analysis in A. anguilla.
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Characterization of pore-forming activity in liver mitochondria from Anguilla anguilla. Two porins in mitochondria?
A fast purification procedure for the isolation and purification of eukaryotic porin (De Pinto et al., (1987) Biochim. Biophys. Acta 905, 499-502) was applied to liver mitochondria of the fish Anguilla anguilla. A protein preparation was obtained which formed slightly anionically selective pores in reconstitution experiments with lipid bilayer membranes. The distribution of single-channel conductances had two maxima of 2.4 nS and 4.0 nS in 1 M KCl. Sodium dodecylsulfate electrophoretograms of the protein preparation showed the presence of two bands of very similar electrophoretic mobility (32 and 32.5 kDa). Both bands cross-reacted with antibodies raised against purified bovine heart porin and with antibodies raised against the 19 amino acids N-terminal end of human porin. No cross-reactivity was observed with antibodies against yeast porin. The peptide maps of the two bands showed slight differences. The possibility of the presence of two different porins in liver mitochondria of Anguilla anguilla is discussed. An extensive immunological comparison of different mitochondrial porins is presented.
Anguilla↗
Anguilla anguilla L. genotoxic and liver biotransformation responses to abietic acid exposure.
Adult eels (Anguilla anguilla L.) were exposed for 8, 16, 24, and 72 h to 0, 0.1, 0.3, 0.9, and 2.7 microM abietic acid (AA). Genotoxicity was measured as erythrocytic nuclear abnormalities (ENA), as well as DNA strand breaks in blood and liver. Liver cytochrome P450 (P450) content, liver ethoxyresorufin O-deethylase (EROD), and glutathione S-transferase (GST) activities were determined as biotransformation biomarkers. Liver alanine transaminase (ALT) activity was also measured as an indication of tissue damage. Low AA concentrations, such as 0.1 and 0.3 microM, result in a delayed induction of A. anguilla L. liver EROD activity, whereas the higher AA concentration (2.7 microM AA) also has a delayed effect probably as a consequence of liver tissue high inhibitory concentration. The current eel liver GST activity results demonstrate that only low AA concentrations promote liver increases in GST, whereas high AA concentrations, such as 0.9 and 2.7 microM, do not alter it. The results concerning eel liver ALT activity indicate that significant liver damage is induced by high AA concentrations, such as 2.7 and 0.9 microM. The eel ENA result analysis reveals that AA is a weak ENA inducer in A. anguilla L. Blood DNA integrity results suggest that low AA concentrations promote late decreases in blood DNA integrity; nevertheless, high AA concentrations are early blood genotoxic inducers compared with low AA doses. According to the present research results with respect to eel liver DNA damage, all of the AA exposure concentrations decreased liver DNA integrity.
Abietanes↗
Anguilla anguilla L. liver EROD induction and genotoxic responses after retene exposure.
Anguilla anguilla L. (European eel) were exposed for 8, 16, 24, and 72 h to 0 (control), 0.1, 0.3, 0.9, and 2.7 microM retene. A. anguilla L. liver ethoxyresorufin-O-deethylase EROD activity significantly increased during the whole exposure period to all retene concentrations, when compared to their controls. The liver cytochrome P450 content only increased after exposure to high retene concentrations (0.9 and 2.7 microM) from 8 to 24 and 72 h, respectively. Generally, liver DNA integrity decreased with increased retene concentrations. Thus, a low retene concentration (0.1 microM) was only effective at 16 h, 0.3 and 0.9 microM had an early and prolonged effect up to 24h, and 2.7 microM decreased liver DNA integrity during the whole exposure period. However, blood DNA integrity decrease was observed in eels after 24h exposure to 0.1 microM retene, and at 16 h to 0.3 and 0.9 microM retene, despite an early blood DNA integrity decrease at 8 and 16 h exposure to 2.7 microM retene. An early genotoxic response to retene was also observed as erythrocyte nuclear abnormalities plus Notched (ENA+Not) frequency increase at 8, 16, and 24h exposure to 0.1 and 0.3 microM retene as well as at 8, 16, 24, and 72 h to 0.9 microM retene. Though, the highest retene concentration (2.7 microM) only induced ENA+Not and erythrocyte nuclear abnormalities minus Notched (ENA-Not) at 16 h exposure. The eel ENA+Not increase was more sensitive than the ENA-Not increase as a measure of retene genotoxicity. Eel liver alanine amino transferase (ALT) increased activity reveals its enhanced transamination capacity after short-term exposure to retene. The A. anguilla L. ratio between hemoglobin concentration and red blood cells count (Hb/RBC) increased at 8h exposure to 0.1, 0.3, and 0.9 microM retene, suggesting an initial homeostatic process.
Alanine Transaminase↗
Endocrine and metabolic changes in Anguilla anguilla L. following exposure to beta-naphthoflavone--a microsomal enzyme inducer.
Anguilla anguilla L. were exposed during 24 and 48 h to 2.7 muM beta-naphthoflavone (BNF), a known microsomal enzyme inducer. The BNF effects on thyroid-stimulating hormone (TSH), free triiodothyronine (T3), free thyroxine (T4) and cortisol plasma levels were investigated. Alterations on plasma glucose and lactate levels were also measured as an indication of energy-mobilizing hormones alterations. BNF showed to be able to decrease significantly A. anguilla plasma T4 levels, whereas TSH, T3 and cortisol plasma remained constant. However, plasma glucose levels were significantly increased, demonstrating that intermediary metabolism has been affected. These results demonstrate that BNF a PAH-like compound alters the normal functioning of the hypothalamo-pituitary-thyroid (HPT) axis in A. anguilla.
Anguilla↗
Anguilla anguilla L. Genotoxic responses after in situ exposure to freshwater wetland (Pateira de Fermentelos, Portugal).
Pateira de Fermentelos is a Cértima River enlargement, close to its river mouth (by the Agueda River), where the introduction of agricultural chemicals such as fertilisers and pesticides, domestic sewage, as well as heavy metals from electroplating industries, results in increased water pollution. The present research work concerns a 48 h in situ exposure of caged eels (Anguilla anguilla L.) at the Pateira de Fermentelos. Five exposure sites were selected, i.e., site A, site B, site C, site D and site E in order to study its water genotoxicity potential, measured in gill, blood, liver and kidney as DNA strand breaks. Eels were also exposed at a reference site by the Cértima River spring. Bottom water samples were collected for further physical-chemical analysis. Site A exposure, significantly decreased gill, blood and liver DNA integrity. Gill and liver DNA integrity was also significantly decreased at site B. At site C only blood DNA integrity was significantly decreased. The present field in situ study demonstrated that the three exposure sites close to the Pateira initial part, such as A, B and C are polluted by pro and/or genotoxic compounds. The genotoxic effects induced in A. anguilla L. suggest a different contamination of the exposure sites A, B and C, in genotoxic chemicals. Thus, according to its genotoxic potential the exposure sites A, B and C, may be ordered as follows site A>site B>site C. No genotoxic effects on A. anguilla L. were observed at site E as DNA strand breaks increase.
Anguilla↗
Mutation analysis of ras gene in the liver of European eel (Anguilla anguilla L.) exposed to benzo[a]pyrene.
Ras is regarded as one of the most important genes involved in carcinogenesis. Such genes have been characterised in several fish species and the presence of ras mutations have already been described in fish populations from hydrocarbon contaminated areas and following experimental exposure to specific contaminants. The aims of this study were to evaluate the DNA integrity by comet assay, to isolate the normal ras gene of Anguilla anguilla and analyse for the presence of ras gene mutations or changes in gene expression levels induced after one month of benzo[a]pyrene (BaP) experimental exposure. The A. anguilla ras cDNA isolated revealed a 189 amino acid protein and alignment with other vertebrate ras proteins revealed conservation of functionally important regions. Following experimental exposure to BaP, an increase in DNA damage was found by comet assay. However, no point mutations or changes in ras gene expression levels were detected when compared to control samples. In contrast to the majority of fish ras gene sequences, a high degree of polymorphic variation was detected in the A. anguilla ras gene.
Amino Acid Sequence↗
Growth differences and growth hormone expression in male and female European eels [Anguilla anguilla (L.)].
In this study, we examined the growth differences of males and females following a sex reversion, and the growth hormone (GH) expression variation between sexes of European eels [Anguilla anguilla (L.)]. A high percentage of females (88%) was found in the group fed with estradiol 17beta compared to the control group (comprised of only 6% female eels), which was defined as the male population. Significant differences between growth rate and size were found following 480 days of growth, whereby the males reached 60+/-4.3 g (means+/-SE) in size and the females 73.4+/-5.9 (g+/-SE); after 600 days, the males reached 114.1+/-4.3 and the females 171+/-11.7 (g+/-SE). A cDNA coding for the complete growth hormone of the European eel A. anguilla (eeGH) was cloned by RACE PCR using several sets of degenerate oligonucleotides. The eeGH cDNA coding region is 627 bp long. A sequence comparison of eeGH with Anguilla japonica GH (jeGH) cDNA showed a 98% identical base. Comparison of the deduced amino acid sequence revealed 99% identical residues, meaning that a difference exists in only two of the 209 residues. In both cases, the differing residues in the eeGH amino acid sequence are lysine. We measured the mRNA levels of growth hormone in the pituitaries of male and female eels growing at different rates. A significantly higher expression of eeGH was found in the female eels in comparison to the males. These results show that different levels of GH transcription eeGH can explain the growth rate differences between male and female European eels.
Administration, Oral↗