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Changes in gas composition in the swimbladder of the European eel (Anguilla anguilla) infected with Anguillicola crassus (Nematoda).

The effect of Anguillicola crassus (Nematoda) on gas composition in the swimbladder of eels (Anguilla anguilla) was studied using mass spectrometry in feral eels from two German rivers and in experimentally infected eels. In both naturally and experimentally infected eels significant correlations were observed between the proportion of oxygen in the swimbladder and level of infection with Anguillicola crassus. In swimbladders of naturally infected feral eels the contribution of oxygen to swimbladder gas was reduced by 36-62.9% and in experimentally infected eels it was reduced by 11.4-57% compared to uninfected controls. The proportion of CO2 appeared to be lower in infected swimbladders compared to uninfected ones. However, this change was not significant. The findings are discussed in relation to an altered structure of the swimbladder wall due to the parasite.

Air Sacs↗

Induction of stress by the swimbladder nematode Anguillicola crassus in European eels, Anguilla anguilla, after repeated experimental infection.

The purpose of this study was to determine under laboratory conditions over a period of 311 days if infection with the nematode Anguillicola crassus induces stress in European eels (Anguilla anguilla), and stimulates the endocrine stress axis as measured by serum cortisol levels. Eels were experimentally infected with 3rd-stage larvae (L3) in different doses to simulate natural conditions with varying infection pressures. Blood samples were drawn from the caudal vein every 2 weeks and serum cortisol concentrations were determined by radioimmunoassay (RIA). The results showed that the application of L3 resulted in a significant increase in the cortisol levels. The period of time at which elevated cortisol values were observed was consistent with the time of larval development and the appearance of adult A. crassus. Thus, there is a stress response to the larval and young adult stages, but no chronic response to older adults. Therefore, it is likely that infection of eels under natural conditions especially with a high number of larvae may be a considerable stressor, in combination with different environmental factors like water temperature, pH, oxygen concentration, pollution and interindividual relationships.

Air Sacs↗

Epidemiology of Anguillicola crassus in the European eel (Anguilla anguilla) from two rivers in southern England.

European eels Anguilla anguilla from the rivers Thames and Test, in the south of England, were examined between 2000 and 2003 for infection with the swim-bladder nematode Anguillicola crassus. Since its introduction to Thames eels at tidal estuarine locations circa 1987, A. crassus has become established in non-tidal freshwater stretches upriver and data from these locations are reported for the first time. The prevalence of infection at Thames estuary locations was higher during 2000-2003 than for the period 1987-1992. By 2003, similar prevalences were observed at freshwater and estuarine locations, but infection intensities were significantly higher in freshwater. Eels from the river Test appear to have been recently colonized by A. crassus (circa 2000). Parasite population establishment within these eels was uncharacteristically slow during 2000-2001, with low prevalence and intensity of infection, and few gravid females during this period. By 2003, infection levels in Test eels were similar to those in Thames eels. The expansion of the A. crassus population in Test eels has occurred in the absence of the paratenic host ruffe Gymnocephalus cernuus, and at suboptimal pH for the survival and infectivity of free-living larvae. The epidemiology of A. crassus in Test eels demonstrates that transmission of A. crassus by ruffe is not required for high prevalence and intensity of infection in eels. However, the consistently low and atypical levels of infection in Test eels during 2000 and 2001 suggests that paratenic transmission by ruffe may provide a substantial contribution to the dynamics of A. crassus in eels in the early years following introduction, by facilitating the rapid increase in prevalence and intensity of A. crassus infection that typically occurs during this time.

Anguilla↗

Physiological status of wild European eels (Anguilla anguilla) infected with the parasitic nematode, Anguillicola crassus.

The effect of the parasitic swimbladder nematode, Anguillicola crassus, on the physiological status of wild European eels (Anguilla anguilla) was investigated during an 18 month survey (February 1995 until September 1996), and compared with that of wild uninfected eels collected over the same time-period. Despite the occurrence of up to 15 blood-feeding adults in the swimbladder lumen of the infected eels and as many as 25 additional larvae in the swimbladder wall, there were no major differences in hormonal, metabolic or osmoregulatory status of the 2 groups of eels. Wild European eels appear to adapt to chronic parasitism with Anguillicola crassus. The possible adverse effects of additional simultaneous stressors are discussed.

Air Sacs↗

Electroretinographic evaluation of spectral sensitivity in yellow and silver eels (Anguilla anguilla).

Although differences in visual pigments between developmental stages of the European eel are well known, the expected differences in spectral sensitivity have not been demonstrated at the electrophysiological level. In fact, one past electroretinographic study led to the conclusion that in eels there is no change in scotopic sensitivity, with increasing sexual maturity. In the present experiments, electroretinograms (ERGs) were recorded from in situ eyecups of immobilized eels Anguilla anguilla (L.) caught in coastal running waters. It was shown that the ERG b-wave is as good an indicator of spectral sensitivity as the unmasked late receptor potential (LRP) which directly reflects the responsiveness of photoreceptors. Complete spectral-sensitivity curves, based on b-wave thresholds and on thresholds of LRP subsequently isolated by means of sodium iodate, have been obtained in the same eel. Using fitted amplitude-log intensity functions for threshold calculation, and two models for computer-assisted fitting of spectral-sensitivity curves, significant differences in lambda max were found between yellow and silver developmental stages of the eel, identified by ocular index measurements.

Anguilla↗

Cloning and expression of three aquaporin homologues from the European eel (Anguilla anguilla): effects of seawater acclimation and cortisol treatment on renal expression.

BACKGROUND INFORMATION: The European eel (Anguilla anguilla) is able to osmoregulate over a wide range of environmental salinities from FW (freshwater) to hyperconcentrated SW (seawater). Successful acclimation is associated with strict regulation of ion and water transport pathways within key osmoregulatory epithelia to enable animals to survive the dehydrating or oedematous conditions. These observations suggested that homologues of the AQP (aquaporin) water channel family were expressed in the eel and that these proteins may contribute to the water transport and osmoregulation in all euryhaline teleosts. RESULTS: Complementary DNAs encoding a homologue of the mammalian aquaglyceroporins (termed AQPe) and two homologues of mammalian aquaporin-1 [termed AQP1 and AQP1dup (aquaporin-1 duplicate)] were isolated from the European eel. Northern-blot analysis revealed (i) two AQP1 transcripts exhibiting a wide tissue distribution, (ii) a single AQP1dup mRNA transcript found in the kidney and the oesophagus, and (iii) a single AQPe mRNA detectable mainly in the kidney and the intestine. The relative expression of isoforms within the kidney was AQP1dup>AQPe>AQP1. SW acclimation significantly reduced the abundance of AQP1, AQP1dup and AQPe transcripts in the kidney of yellow eels by approx. 72, 66 and 34% respectively, whereas the expression levels in silver eels were independent of salinity and equivalent to those observed in yellow SW-acclimated fish. AQP1 protein expression was primarily located within the vascular endothelium in yellow eels and the epithelial apical brush border in some renal tubules in silver eels. Infusion of cortisol into FW eels had no effect on AQPe mRNA expression, but induced significant decreases in AQP1 and AQP1dup mRNA levels in the kidney of yellow eels. Cortisol infusion had no effect on the expression of any isoform in the silver eels. CONCLUSIONS: These results suggest that SW-acclimation or cortisol infusion induces a down-regulation of renal AQP expression in yellow eels. However, the lower levels of aquaporin expression found within the silver eel kidney were not further reduced by salinity transfer or steroid infusion. These differences in mRNA expression were accompanied by changes in the cellular distribution of the AQP1 protein between vascular endothelial and tubular epithelial cells.

Acclimatization↗

The involvement of cytochrome P450 system in the fate of 2,4,6-trinitrotoluene (TNT) in European eel [Anguilla anguilla (Linnaeus, 1758)].

TNT (2,4,6-trinitrotoluene) was the most common nitro aromatic explosive available in World War II ammunitions. The presence of ordnance dumped at sea might represent a great concern for marine species living close to dumping sites and the toxicological properties of the chemicals released into the marine environments need to be evaluated. The aim of the present study is to investigate the involvement of CYP (cytochrome P450) system in the metabolism of TNT in marine organisms by using the European eel [Anguilla anguilla (Linnaeus, 1758)] as model species. In vivo exposure to sublethal concentration of TNT (0.5, 1 and 2.5 mg/l) leads to a significant decrease in the phase I CYP1A catalytic activities such as EROD (7-ethoxyresorufin-O-de-ethylase) and MROD (7-methoxyresorufin-O-de-ethylase). On the opposite, a significant increase in NADPH cytochrome c reductase activity as well as phase II UDP-glucuronosyltransferase activity is observed. An inhibition at enzyme level is hypothesized for both CYP1A enzymes, also confirmed by a similar decrease observed after in vitro exposure. An active role of NADPH cytochrome c reductase and phase II enzymes in the TNT metabolism may also be hypothesized.

Anguilla↗

Localization of carbonic anhydrase in swimbladder of European eel (Anguilla anguilla) and perch (Perca fluviatilis).

The distribution of carbonic anhydrase in swimbladder tissue and especially in gas gland cells of the European eel (Anguilla anguilla) and the perch (Perca fluviatilis) was analysed using histochemical staining according to Hansson (1967), with modifications proposed by Riddersträle (1991). While in the European eel, gas gland cells are distributed as a single layered epithelium over the whole secretory part of the swimbladder, the gas gland of the perch consists of a compact, richly vascularized 'multilayered' epithelium, in which gas gland cells have contact with the swimbladder lumen via small channels. In spite of these differences in organization, membranes of gas gland cells near blood vessels are richly folded in both species. A strong histochemical staining for carbonic anhydrase was observed in these membrane foldings. With prolonged incubation times a positive reaction was also observed in the cytoplasm of gas gland cells. In addition, the vascular endothelium and the erythrocytes showed a positive histochemical reaction. No staining, however, was visible in apical membranes towards the lumen of the swimbladder. In the perch, swimbladder epithelial cells outside the gas gland showed no positive staining of carbonic anhydrase. The results thus indicate that carbonic anhydrase activity is especially concentrated in membranes facing blood vessels. This suggests that a rapid equilibrium of the CO2/HCO3- reaction in the intracellular as well as in the extracellular space is essential for swimbladder function.

Air Sacs↗

Evidence that corticotropin-releasing hormone acts as a growth hormone-releasing factor in a primitive teleost, the European eel (Anguilla anguilla).

The inhibitory control of growth hormone (GH) release by somatostatin (SRIH) has been conserved throughout vertebrate evolution. In contrast, the neuropeptides involved in the stimulatory control of GH vary according to species and/or physiological situations. We investigated the direct pituitary regulation of GH release in a primitive teleost, the European eel (Anguilla anguilla L.) at the juvenile stage. Short-term serum-free primary cultures of dispersed pituitary cells were used, and GH release was measured by an homologous radioimmunoassay. Whereas growth hormone-releasing hormone (GHRH), gonadotropin-releasing hormone (GnRH), thyrotropin-releasing hormone (TRH), neuropeptide Y (NPY) and cholecystokinin (CCK) failed to induce any change in GH release, corticotropin-releasing hormone (CRH) dose-dependently stimulated GH release with a significant effect at 1 nM and a maximal effect (> or =400% of controls at 24 h) at 100 nM. In agreement with our previous studies, PACAP also stimulated GH release but its maximal effect was lower than that of CRH. Proopiomelanocortin (POMC)-peptides, corticotropin (ACTH), melanotropin (alpha-MSH), beta-endorphin) had no effect on GH release, at any dose tested (0.1-1000 nM), indicating that the stimulatory effect of CRH on GH release by somatotrophs was not mediated by CRH-induced release of POMC-peptides from corticotrophs and melanotrophs. The CRH antagonist, alpha-helical CRH(9-41), significantly inhibited the stimulatory effect of CRH on GH release, suggesting the implication of specific CRH receptors related to mammalian ones. The stimulatory effect of CRH on GH release was reduced after 24 h of incubation, indicating a desensitization. In contrast, no desensitization to the inhibitory effect of SRIH was observed. SRIH inhibited CRH action in a dose-dependent manner. The effect of SRIH was overriding, 1 nM SRIH being able to abolish the effect of 1000 nM CRH. In conclusion, in the eel, CRH stimulates GH release directly at the pituitary cell level. GH and cortisol secretions could interact in controlling several physiological functions such as metabolism and ion exchange. This study suggests that CRH may have played an important early role in vertebrates co-ordinating the activation of various endocrine axes involved in metamorphosis, osmoregulation, stress and fasting. The stimulatory role of CRH on GH release may have been partially conserved during evolution, as it is found in some human physio-pathological situations such as stress, fasting and depression.

Adrenocorticotropic Hormone↗

The anodic hemoglobin of Anguilla anguilla. Molecular basis for allosteric effects in a root-effect hemoglobin.

The functional and structural basis for the Root effect has been investigated in the anodic hemoglobin of the European eel, Anguilla anguilla. This hemoglobin exhibits a large Bohr effect, which is accounted for by oxygen-linked binding of seven to eight protons in the presence of GTP at pH 7.5. Oxygen equilibrium curves show nonlinear lower asymptote of Hill plots, indicating the occurrence of heme-heme interactions within the T state. Analysis of the curves according to the co-operon model (Brunori, M., Coletta, M., and Di Cera, E. (1986) Biophys. Chem. 23, 215-222) reveals that T state cooperativity is positive at high pH and in the stripped hemoglobin (where the T --> R allosteric transition is operative) and negative at low pH and in the presence of organic phosphate (where the molecule is locked in the low affinity structure), indicating site heterogeneity. The complete amino acid sequence of eel anodic hemoglobin has been established and compared with that of other fish hemoglobins. The presence of the Root effect correlates with a specific configuration of the alpha1beta2 switch interface, which at low pH would stabilize subunit ligation in the T state without changing the quaternary structure. We propose that the major groups involved in the binding of oxygen-linked protons in eel anodic hemoglobin are located on the beta chain and comprise His-HC3 at the C terminus, His-FG4 at the switch interface, and Lys-EF6 and the N terminus at the phosphate-binding site.

Amino Acid Sequence↗

Early emergence of three dopamine D1 receptor subtypes in vertebrates. Molecular phylogenetic, pharmacological, and functional criteria defining D1A, D1B, and D1C receptors in European eel Anguilla anguilla.

The existence of dopamine D1C and D1D receptors in Xenopus and chicken, respectively, challenged the established duality (D1A and D1B) of the dopamine D1 receptor class in vertebrates. To ascertain the molecular diversity of this gene family in early diverging vertebrates, we isolated four receptor-encoding sequences from the European eel Anguilla anguilla. Molecular phylogeny assigned two receptor sequences (D1A1 and D1A2) to the D1A subtype, and a third receptor to the D1B subtype. Additional sequence was orthologous to the Xenopus D1C receptor and to several other previously unclassified fish D1-like receptors. When expressed in COS-7 cells, eel D1A and D1B receptors display affinity profiles for dopaminergic ligands similar to those of other known vertebrate homologues. The D1C receptor exhibits pharmacological characteristics virtually identical to its Xenopus homologue. Functionally, while all eel D1 receptors stimulate adenylate cyclase, the eel D1B receptor exhibits greater constitutive activity than either D1A or D1C receptors. Semiquantitative reverse transcription-polymerase chain reaction reveals the differential distribution of D1A1, D1A2, D1B, and D1C receptor mRNA within the hypothalamic-pituitary axis of the eel brain. Taken together, these data suggest that the D1A, D1B, and D1C receptors arose prior to the evolutionary divergence of fish and tetrapods and exhibit molecular, pharmacological, and functional attributes that unambiguously allow for their classification as distinct D1 receptor subtypes in the vertebrate phylum.

Adenylyl Cyclases↗

Helminth communities in eels Anguilla anguilla from Adriatic coastal lagoons in Italy.

The composition and diversity of the total and intestinal component and infra-communities were determined in eels Anguilla anguilla from three shallow lagoons on the Adriatic coast of Italy to determine whether the helminth communities would differ in composition and structure from those in eels from lagoons on the Tyrrhenian coast. The lagoons differed in respect of their management regimes and the extent of freshwater influx. Both freshwater and marine species of helminths were found in the eels in all three lagoons, but the freshwater component was richer in Valle Figheri. A suite of three digenean eel specialist species occurred in all three lagoons, of which any two members dominated each community. This conferred a high degree of similarity between the communities of the three lagoons. The same three species also dominated helminth communities in eels in lagoons along the Tyrrhenian coast of Italy, and compositional similarity levels were similar within and between western and eastern groups. Species richness was higher in the component communities of the eels of the Adriatic lagoons when compared to the Tyrrhenian ones, but diversity and dominance indices were of a similar order of magnitude and range. Intestinal helminth communities were richer and more diverse in two of the Adriatic lagoons because the proportion of eels with zero or one helminth species was, unusually, in the minority. It was nevertheless concluded that infracommunity structure was similar in eels from both western and eastern lagoons and that the hypothesis that it would differ in Adriatic lagoons could not be supported. The findings provide further evidence of the similarity in composition and structure of helminth communities in eels from coastal lagoons throughout Europe.

Anguilla↗

Patterns of infracommunity species richness in eels, Anguilla anguilla.

Between October 1999 and October 2001, a total of 510 European eels Anguilla anguilla were captured in 13 different samples from the rivers Thames (five locations) and Test (one location) in southern England. The relationship between parasite component community species richness (CCR) and maximum infracommunity species richness (ICRmax) compared with that previously observed in bird and mammal hosts. Specifically, the maximum number of parasite species occurring in infracommunities equalled or exceeded half the number of parasite species in the component community at that time, across a wide range of CCR values (2-9 parasite species). Furthermore, the frequency distribution of infracommunity richness (ICR) suggested that the species composition of infracommunities is probably random. These findings suggest that intestinal macroparasite infracommunities in eels are unsaturated and potentially species rich assemblages and, in these respects, share a fundamental similarity with the infracommunities of birds and mammals.

Anguilla↗

Accumulation of the precious metals platinum, palladium and rhodium from automobile catalytic converters in Paratenuisentis ambiguus as compared with its fish host, Anguilla anguilla.

The platinum group metals (PGM) Pt, Pd and Rh are emitted into the environment mainly by catalytic exhaust gas converters of cars. As PGM accumulate in sediments of aquatic ecosystems, the study was focused on the uptake of the noble metals by European eels, Anguilla anguilla infected with the acanthocephalan Paratenuisentis ambiguus. Eels were exposed to ground catalytic converter material for six weeks. After exposure Pt and Pd were detected in the liver and kidney of the eels and in the parasites. Palladium was also found in fish muscle and intestine. No Rh uptake by the eel tissues and the parasites occurred. Paratenuisentis ambiguus contained the highest levels of both metals with 40 times higher Pt concentrations and four times higher Pd concentrations than the liver of its host. Due to its accumulation capacity for PGM, P. ambiguus can be applied as a sensitive accumulation indicator in field studies to assess the degree of environmental PGM contamination in aquatic ecosystems.

Acanthocephala↗

Lethal toxicity of Lindane on a teleost fish, Anguilla anguilla from Albufera Lake (Spain): hardness and temperature effects.

This paper reports the results of toxicity tests conducted using Anguilla anguilla under three different water temperature (15, 22 and 29 degrees C) and two hardness regimes (250 and greater than 600 ppm CaCO3). The 96-h LC50 increased in the experimental medium (p less than 0.05) by an order of magnitude from 0.32 to 0.45 mg/L between 15 and 29 degrees C. However in the natural medium it is similar (p greater than 0.05) (0.54 to 0.55 mg/L) for these same temperatures. The toxicity of Lindane on eels increased when the water hardness decreased. The 24, 48, 72 and 96-h LC50 for this fish in both media is less at 15 degrees C (96-h LC50 = 0.32 and 0.55 mg/L) than at 29 degrees C (96-h LC50 = 0.45 and 0.55 mg/L). These results suggest that the toxicity of Lindane presents a negative temperature coefficient.

Anguilla↗

Comparative acute toxicities of selected pesticides to Anguilla anguilla.

The acute toxicities (24, 48, 72 and 96 hr) of eight pesticides to Anguilla anguilla were determined. The organochlorine pesticide, endosulfan was the most toxic, with LC50 values in the range of 0.042 to 0.041 mg/L. Endosulfan was followed in order of decreasing toxicity by diazinon, fenitrothion, chlorpyrifos, lindane, methidathion, trichlorfon and methylparathion. When fishes were exposed to the pesticides tested they exhibited signs of restlessness, erratic swimming, convulsions and difficulty in respiration. This response was more persistent in fishes exposed to organophosphorus pesticides.

Anguilla↗

Acute toxicity, uptake and clearance of diazinon by the European eel, Anguilla anguilla (L.).

The toxicity, accumulation, and elimination of diazinon were investigated for the european eel, Anguilla anguilla. The 24, 48, 72 and 96-h median lethal concentrations (LC50) were 0.16, 0.11, 0.09 and 0.08 mg/L, respectively. Fish exposed to sublethal concentration (0.042 mg/L) accumulated diazinon in liver and muscle tissues. Bioconcentration factors (BCF) of diazinon were 1850 in liver, and 775 in muscle over the 96-h exposure period. Upon removal from diazinon containing water the contaminated fish rapidly eliminated diazinon. The excretion rate constants of this insecticide were 0.108 h-1 for liver and 0.016 h-1 for muscle. Diazinon half-lives were 16.6 and 33.2 hours for liver and muscle, respectively.

Anguilla↗

Physiological stress responses of Anguilla anguilla to fenitrothion.

The present communication deals with the effect of fenitrothion (0.04 ppm) on the carbohydrate metabolism of the european eel Anguilla anguilla and its recovery from intoxication. Analysis of various parameters such as glycogen, lactate and glucose was made on different eel tissues after 2, 8, 12, 24, 32, 48, 56, 72 and 96 hr of fenitrothion exposure. Subsequently, the fish were allowed recovery periods of 8, 12, 24, 48, 72, 96, 144 and 192 hr in clean water, and the same parameters were evaluated. Muscle glycogen content decreased significantly during the exposure time, while blood glucose levels increased markedly. Gill, muscle and blood lactate values increased during pesticide exposure. Most of the metabolic disorders did not persist after allowing recovery in clean water during less than a week. The observed effects of fenitrothion on fish metabolism are discussed in relation to a stress syndrome and probable reasons for alterations have been exposed.

Analysis of Variance↗