Dynamics and predicted decline of Anguillicola crassus infection in European eels, Anguilla anguilla, in Neusiedler See, Austria

The eel population in Neusiedler See has been maintained by regular massive stocking since 1958. After the establishment of the National Park Neusiedler See-Seewinkel in 1993, eel stocking was prohibited and the population, together with the specific parasites of eels, was predicted to decline to extinction within 10 years. This investigation was undertaken to document the decline and extinction of the Anguillicola crassus population in eels. From 1994 to 2001, 720 eels were collected from two sites in the lake. Prevalence and abundance of A. crassus were lower in spring than in summer and autumn and larger eels harboured more parasites than smaller ones. Neither year of study nor sampling site were correlated with parasite infection levels. No significant trend in the population parameters of A. crassus was detected over the 8 years of the survey. This suggested that there had been no significant decline in the eel population. This suggestion was confirmed by investigations of the fishery, which also found evidence of regular illegal stocking. The stability of the A. crassus population over the past decade seems to reflect the lack of change in eel population density. No mass mortalities of eels occurred over the period despite the many similarities between Neusiedler See and Lake Balaton in Hungary. Differences in eel size, eel diet and the lack of large-scale insecticide use are discussed as possible explanations for the absence of eel mass mortalities in Neusiedler See.     

Antidiuretic Effect of Eel ANP Infused at Physiological Doses in Conscious, Seawater-Adapted Eels, Anguilla japonica

Atrial natriuretic peptide (ANP) is known as a potent natriuretic/diuretic hormone in vertebrates. However, eel ANP infused at doses that did not alter arterial blood pressure (0.3-3.0 pmol/kg/min) decreased urine volume and increased urinary Na concentration in seawater (SW)-adapted eels but not in freshwater (FW)-adapted eels. The renal effects were dose-dependent and disappeared after infusate was switched back to a vehicle (0.9% NaCl). Urinary Na excretion (volume x Na concentration) did not change during ANP infusion. ANP infusion increased plasma ANP concentration, but the increase at the highest dose was still within those observed endogenously after injection of hypertonic saline. Urinary Mg and Ca concentrations increased during ANP infusion in SW eels, but urinary Ca excretion decreased in FW eels. Plasma Na concentration profoundly decreased during ANP infusion only in SW eels, suggesting that ANP stimulates Na extrusion via non-renal routes. These results indicate that ANP is a hormone which specifically extrudes Na ions and thereby promotes SW adaptation in the eel. This is in sharp contrast with mammals where ANP is a volume regulating hormone that extrudes both Na and water.     

Polycyclic aromatic hydrocarbons and heavy metals in Kostrena coastal area

The aim of this study was to determine pollution by polycyclic aromatic hydrocarbons (PAH) and heavy metals in seawater and sediment in Kostrena coastal area, as well as their toxicity using bioluminescence based tests. Total PAH concentration in seawater ranged 1.7-155.3 ng/L. The share of carcinogenetic PAH was relatively high, ranging 22-48.3%. Nickel concentrations in seawater were beyond detection limits (< 0.1 microg/L), vanadium concentrations ranged 0.66-1.96 microg/L, chrome concentrations were beyond detection limits, and copper concentrations were also beyond detection limits or extremely low (up to 0.32 microg/L). EC50 values in seawater ranged 23.80-90.90 ng/L. Correlation between total PAH concentration and toxicity of seawater showed strong connection between them (r = 0.9579). Total PAH concentration in marine sediment ranged 58.02-1116 microg/kg dry weight (d.w.). The share of carcinogenetic PAH was extremely high ranging 10-53%. Nickel concentrations in marine sediment ranged 8-24 mg/kg d.w., vanadium concentrations ranged 24-42 mg/kg d.w., chrome concentrations ranged 11-19 mg/kg d.w., and copper concentrations ranged 7-25 mg/kg d.w. EC50 values in marine sediment ranged 818-4596 microg/kg d.w. Correlation between total PAH concentration and toxicity of marine sediment showed weak connection between them (r = 0.2590). Previous studies of seawater samples from areas of the Adriatic sea under the direct influence of oil industry did not include concentrations of heavy metals, which makes our study the first to present such comprehensive results. Our results point out the need for further evaluations and following of marine environment pollution and its consequences on living organisms and marine ecosystem in whole.     

American eels produce and release bile acid profiles that vary across life stage

The American eel (Anguilla rostrata) is an imperilled fish hypothesized to use conspecific cues, in part, to coordinate long-distance migration during their multistage life history. Here, holding water and tissue from multiple American eel life stages was collected and analysed for the presence, profile and concentration of bile acids. Distinct bile acid profiles were identified in glass, elver, yellow eel and silver eel holding waters using ultraperformance liquid chromatography high-resolution mass spectrometry and principal component analysis. Taurochenodeoxycholic acid, taurodeoxycholic acid, cholic acid, deoxycholic acid, taurolithocholic acid and taurocholic acid were detected in whole tissue of American glass eels and elvers, and in liver, intestine and gallbladder samples of late-stage yellow eels. Bile acids were not a major component of silver eel washings or tissue. This study is novel because little was previously known about bile acids produced and emitted into the environment by American eels. Future behavioural studies could evaluate whether any bile acids produced by American eels influence conspecific migratory behaviour.     

Callus tissue of Solanum tuberosum L. cultured in the presence of the pyrethroid deltamethrin

The effect of Decis (deltamethrin as active ingredient) on callus tissue of Solanum tuberosum L. cv. Désirée was studied. Decis is an agrochemical currently used on field grown potato plants to control the Colorado beetle, a potato pest. Deltamethrin added to the culture medium interferes with the behaviour of callus tissue. After 5 h of culture, the level of total proteins was higher in treated tissue than in the control, and SDS-PAGE showed that deltamethrin promoted the increase of some soluble proteins. After 24 h and 14 days of culture, the level of total proteins became similar in both treated and control material. This similarity between control and treated tissues, after 14 days of culture, occurred as a result of the treatment with deltamethrin which caused a decrease of soluble proteins, but an increase in insoluble proteins whereas the opposite was observed for control callus. SDS-PAGE of both soluble and insoluble proteins showed that only quantitative dissimilarities occurred. This longer treatment also increased the chlorophyll content. Ultrastructural study of the cells revealed that tissue callus cultured for 14 days in the presence of deltamethrin had plastids containing a more developed membranous system, with a higher number of grana and with more compartments than in control cells. Deltamethrin also promoted the abundance of vesicles associated with dictyosomes. This response of the callus tissue to Decis added to the callus medium parallels the behaviour of potato plants treated with this agrochemical under field conditions (Fidalgo, Santos & Salema, 1993).     

Characterization of Anopheles minimus CYP6AA3 expressed in a recombinant baculovirus system

Metabolism by cytochrome P450 monooxygenases is a major mechanism implicated in resistance of insects to insecticides, including pyrethroids. We previously isolated the cytochrome P450 CYP6AA3 from deltamethrin-selected resistant strain of Anopheles minimus mosquito, a major malaria vector in Thailand. In the present study, we further investigated the role of CYP6AA3 enzyme in deltamethrin metabolism in vitro. The CYP6AA3 was expressed in Spodoptera frugiperda (Sf9) insect cells via baculovirus-mediated expression system. The enzymatic activity of CYP6AA3 in deltamethrin metabolism was characterized after being reconstituted with An. minimus NADPH-cytochrome P450 reductase and a NADPH-regenerating system. The contribution of CYP6AA3 responsible for deltamethrin metabolism was determined by measurement of deltamethrin disappearance following the incubation period and deltamethrin-derived compounds were detected using combined gas chromatography mass spectrometry analysis. 3-Phenoxybenzaldehyde was a major product of CYP6AA3-mediated deltamethrin metabolism. Deltamethrin degradation and formation of metabolites were NADPH-dependent and inhibited by piperonyl butoxide. Deltamethrin was catalyzed by CYP6AA3 with an apparent K(m) of 80.0 +/- 2.0 and V(max) of 60.2 +/- 3.6 pmol/min/pmol P450. Furthermore, deltamethrin cytotoxicity assays by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue dye exclusion were examined in Sf9 insect cells, with and without expression of CYP6AA3. Results revealed that CYP6AA3 could play a role in detoxifying deltamethrin in the cells. Thus, the results of this study support the role of CYP6AA3 in deltamethrin metabolism.     

Biological and chemical assays of pyrethroids in cattle dung

Bioassays were developed in Zimbabwe to measure pyrethroid in cattle dung. These and chemical assays then estimated concentrations in dung from treated oxen and elucidated risks to dung fauna. Laboratory bioassays with adult beetles (Histeridae and Scarabaeinae, including Copris, Digitonthophagus, Onitis and Sisyphus spp.) and muscoid larvae (Musca lusoria Wiedemann) indicated that the LC50 of pyrethroids, as ppm in the wet weight, averaged 0.04 for deltamethrin pour-on, 0.25 for deltamethrin dip, 0.22 for alphacypermthrin pour-on, 0.10 for cyfluthrin pour-on, 0.23 for cypermethrin dip and 0.63 for flumethrin dip. Field bioassays involved artificial dung pats of 800 g, deployed in woodland and inspected after 24 h to record insects dead and alive. Beetles were most abundant in the wet season. Muscoid larvae were less seasonal. The LC50 of insecticides in the field confirmed laboratory indications. Adult Diptera (muscoids and Sgifidae) were not repelled or killed until the deltamethrin concentration reached 10 ppm. Pat dispersal by dung fauna and termites (Microtermes spp.) was halved by deltamethrin at 0.1-1 ppm. Scavenging of dead beetles by ants was greatest with small beetles (< 15 mm long) uncontaminated with insecticide. Dips and pour-ons of deltamethrin on cattle gave residues of about 0.01-0.1 ppm in dung produced in the fortnight after application. About 1.6% of the deltamethrin applied was transferred to dung. Deltamethrin and alphacypermethrin in dung showed no detectable degradation in 64 days. Contamination levels threaten populations of slow-breeding beetles.     

溴氰菊酯对不同品系家蝇脑突触体膜蛋白磷酸化及ATP酶活性的影响

以敏感品系家蝇和溴氰菊酯抗性品系家蝇(Musca domestica L.)为材料,研究和比较了神经毒剂溴氰菊酯对其脑突触体蛋白磷酸化作用的影响。结果表明,浓度为10^-5 mol/L溴氰菊酯抑制了敏感品系家蝇脑突触体蛋白磷酸化作用,而对抗性品系家蝇脑突触体蛋白磷酸化作用无明显影响。若反应体系中加入2.5×10^-6 mol/L的cAMP显著激活了敏感品系家蝇脑突触体蛋白磷酸化水平,但是当0.6 mmol/Lca²⁺或0.6 mmol/L Ca²⁺加10^-5 mol/L钙调蛋白时明显增强了抗性品系家蝇脑突触体蛋白磷酸化水平,甚至超过了其对敏感品系的作用水平。此外,还发现不同浓度的溴氰菊酯可抑制突触膜上的Na/K-ATP酶和Ca-ATP酶活力,浓度越高抑制作用也越大,并且敏感品系家蝇对溴氰菊酯的敏感度要高于抗性品系。     

Regulation of expression of two aquaporin homologs in the intestine of the European eel: effects of seawater acclimation and cortisol treatment

Complementary DNAs encoding homologs of the mammalian aquaglyceroporins (termed AQPe) and aquaporin-1 isoforms (termed AQP1) were isolated from the European eel. The AQP amino acid sequences share 35-54% identity with other known human AQPs. Although AQPe mRNA expression was approximately equivalent along the entire length of the gut, AQP1 expression was the highest in the posterior/rectal segment. Seawater (SW) acclimation increased AQP1 mRNA abundance by 5- and 17-fold in the anterior, 14- and 23-fold in the mid-, and 9- and 7-fold in the posterior/rectal gut regions of yellow and silver eels, respectively. SW acclimation had an effect on AQPe mRNA expression only in the midintestine of silver eels, where a small but significant 1.7-fold increase in abundance was measured. Western blots using an eel AQP1-specific antibody identified the presence of a major immunoreactive 28-kDa protein, primarily within the posterior/rectal segment. A 3-wk SW transfer induced an increase in AQP1 protein abundance in all intestinal segments, with the posterior/rectal region still expressing protein levels approximately 40- and 8-fold higher than the anterior and midsegments, respectively. Strong AQP1 immunofluorescence was detected within the vascular endothelium in both freshwater (FW)- and SW-acclimated eels and in the epithelial apical brush border in the posterior/rectal gut regions of SW-acclimated eels. Cortisol infusion into FW eels had no effect on intestinal AQPe mRNA expression but induced increases in AQP1 mRNA and protein levels. These results provide evidence for the presence of a SW-induced and steroid-regulated AQP water channel pathway within the intestine of the European eel.     

Relative abundance, sex ratio and population structure of the Japanese eel Anguilla japonica in the Tanshui River system of northern Taiwan

From the Gong-Shy-Tyan (GST) Stream and the Tanshui River, Taiwan, eels ranged from 5.53 cm t.l . (elvers) to 72.5 cm t.l . Anguilla japonica accounted for 93-99%, A. marmorata 1-7% and A. bicolor pacifica less than 1% of all eels caught. Mean eel lengths increased from 10 cm t.l . at the downstream sites to 50 cm t.l . at the upstream sites. Females made up 92.8% of the sex-determined eels. Population density, averaged approximately 0.14 eel m⁻² (2.42 g m⁻²) and 0.25 eel m⁻² (0.92 g m⁻²) in downstream sites of the GST and Tanshui River, respectively, and decreased substantially with upstream distance. Eels were rarely found in the heavily polluted and dam constructed areas in the midstream site of the Tanshui River.     

Differential expression of neural and gonadal aromatase enzymatic activities in relation to gonadal development in Japanese eel, Anguilla japonica

The objectives of the present study were to investigate the enzymatic characteristics of brain aromatase in Japanese eel, Anguilla japonica, as well as the correlations between aromatase activities in various tissues (brain regions, pituitary, and gonads) and ovarian development. Eel brain aromatase exhibited a K(m) of 75 nM and a V(max) of 1.14 fmol/min mg protein (91.5 fmol/h mg protein), indicating an enzymatic activity much lower than in other teleosts but similar to that in mammals. This supports the hypothesis of the occurrence of a single aromatase gene in eels (representative of an ancient group of teleosts, Elopomorphs), as in mammals, but unlike what is observed in more recent teleosts. Aromatase inhibitors could significantly suppress brain and pituitary aromatase activity. There was no significant sex difference in aromatase activity in the forebrain, midbrain, hindbrain, or pituitary, but there was in the gonads, where aromatase could be detected in the ovaries but not in the testes, in accordance with the role of this enzyme in ovarian differentiation. Comparison with another teleost, black porgy (Percomorph), under the same experimental conditions, further confirmed the low activity of aromatase in the eel. We investigated variations in brain, pituitary, and gonad aromatase activity in relation to ovarian development in control female eels (gonadosomatic index, GSI, 0.1-1.6%) as well as in eels treated with pituitary extract (experimental ovarian maturation; GSI up to 25%). Differential expression of neural and gonadal aromatase was observed in relation to the course of gonadal development. Pituitary aromatase activity increased with GSI at all stages. Brain (specially forebrain) aromatase activity significantly increased in early vitellogenic control eels (GSI>0.8%) and in treated eels. The low activity of eel aromatase may be related to the characteristic life cycle of the eel, in which there is a long delay of the onset of puberty before oceanic reproductive migration.     

A model to predict the presence of longfin eels in some New Zealand streams, with particular reference to riparian vegetation and elevation

Two logistic regression models were developed from a database of 27 biotic and physicochemical variables for 99 sites in the Taieri River, New Zealand, to predict the probability of occurrence of longfin eels. Average depth was associated positively with eels while woody debris and oxygen concentration were negatively associated. At a macro-scale probability of eel occurrence declined with increasing elevation and, for a given elevation, was higher in tussock and pasture catchments and lower in pine and native forest settings. Using a separate fish database for the Taieri River this macro-scale model predicted eel presence 95·4% in agreement with observation. A map was generated from the model showing areas of predicted high, moderate and low probabilities of eel occurrence. The model also estimated the minimum total number of eels present in the Taieri River catchment (excluding lakes, and streams below 100 m and above 1000 m) as 20 865 (95% CL: 10 560-36 350).     

The eels of Lake Kariba: distribution, taxonomic status, age, growth and density

It had been predicted that eels would disappear from Lake Kariba because juveniles would be unable to surmount the dam which was closed in 1958. This prediction has been proved wrong. In 1970–71 an abundant population of eels was discovered in the lake, mostly at depths from 25–40 m. The eels have intermingled characters of both the mottled Anguilla nebulosa labiata and the plain-colored A. mossambica . A clear separation of these two taxa was impossible and therefore conspeciflc status of these two species is suspected; the name A. n. labiata is retained for the population.
The age structure of the eel cohorts suggests that juveniles surmount the dam in their second year of life and then spend approximately 7 years in streams of the lake drainage. The fastest maturing individuals which are in best condition then emigrate to the ocean, whereas the slower maturing eels remain in the lake longer. The oldest eel in the samples was 18 years old.
The catch per unit of effort for hoopnets was 2.35 kg in the upper part of the lake and 0.41 kg for the lower part of the lake. The density at Namazambwe was assessed at 46 eels per 1 ha. The unexploited eel population of Lake Kariba could form a valuable resource.     

Enhanced expression and release of C-type natriuretic peptide in freshwater eels

C-type natriuretic peptide (CNP) is recognized as a paracrine factor acting locally in the brain and periphery. To assess the role of CNP in teleost fish, a cDNA encoding a CNP precursor was initially cloned from the eel brain. CNP message subsequently detected by ribonuclease protection assay, using the cDNA as probe, was most abundant in the brain followed by liver, gut, gills, and heart. Expression was generally higher in freshwater (FW) than in seawater (SW) eels, but not in the brain. Plasma CNP concentration measured by a newly developed homologous radioimmunoassay for eel CNP was higher in FW than in SW eels. The CNP concentration was also higher in the heart of FW eels but not in the brain. These results show that CNP is abundantly synthesized in peripheral tissues of FW eels and secreted constitutively into the circulation. Therefore, CNP is a circulating hormone as well as a paracrine factor in eels. Together with our previous demonstration that CNP-specific receptor expression is enhanced in FW eels, it appears that CNP is a hormone important for FW adaptation. Because atrial NP (ANP) promotes SW adaptation in eels, CNP and ANP, despite high sequence identity, appear to have opposite effects on environmental adaptation of the euryhaline fish.     

Genotoxic effects of deltamethrin in the mouse bone marrow micronucleus assay

The genotoxicity of deltamethrin was studied in Swiss albino male mice (five animals/group) using the bone marrow micronucleus assay. Deltamethrin (two i.p. injections, 30 h and 6 h before sample collection) was found to induce micronuclei at 162.5 and 300.0 mg/kg body weight. A lower dose (32.5 mg/kg body weight) failed to induce a significant increase in micronuclei over the control level.     

Roles of Slc13a1 and Slc26a1 sulfate transporters of eel kidney in sulfate homeostasis and osmoregulation in freshwater

Sulfate is required for proper cell growth and development of all organisms. We have shown that the renal sulfate transport system has dual roles in euryhaline eel, namely, maintenance of sulfate homeostasis and osmoregulation of body fluids. To clarify the physiological roles of sulfate transporters in teleost fish, we cloned orthologs of the mammalian renal sulfate transporters Slc13a1 (NaSi-1) and Slc26a1 (Sat-1) from eel (Anguilla japonica) and assessed their functional characteristics, tissue localization, and regulated expression. Full-length cDNAs coding for ajSlc13a1 and ajSlc26a1 were isolated from a freshwater eel kidney cDNA library. Functional expression in Xenopus oocytes revealed the expected sulfate transport characteristics; furthermore, both transporters were inhibited by mercuric chloride. Northern blot analysis, in situ hybridization, and immunohistochemistry demonstrated robust apical and basolateral expression of ajSlc13a1 and ajSlc26a1, respectively, within the proximal tubule of freshwater eel kidney. Expression was dramatically reduced after the transfer of eels from freshwater to seawater; the circulating sulfate concentration in eels was in turn markedly elevated in freshwater compared with seawater conditions (19 mM vs. 1 mM). The reabsorption of sulfate via the apical ajSlc13a1 and basolateral ajSlc26a1 transporters may thus contribute to freshwater osmoregulation in euryhaline eels, via the regulation of circulating sulfate concentration.     

Radiodiagnostic method for studying the dynamics of Anguillicola crassus (Nematoda: Dracunculoidea) infection and pathological status of the swimbladder in Lake Balaton eels

Swimbladder changes resulting from Anguillicola crassus infection of the European eel Anguilla anguilla have been the subject of several studies reported in the literature. These investigations, however, studied exclusively the status of infection at a given point in time and did not deal with changes in swimbladder infection in eels suffering from anguillicolosis over a period of time. In this study, A. crassus-induced pathological changes were monitored in 78 eels naturally infected in Lake Balaton and subsequently kept in the laboratory, thus excluding the possibility of further infection. During the 3 mo study, the status of the swimbladder was checked by radiographic examination on 4 occasions. At the end of the study the eels were dissected and the gross pathological changes in the swimbladders were compared with the radiographic findings. As compared to their starting condition, by the end of the study the pathological status of the swimbladder had deteriorated in 55% and remained the same in 37% of the cases. Tendency to improvement (1%) and variable findings (7%) were recorded in a low percentage of cases only. With the help of the radiographs presented, the dynamics of A. crassus infection and of changes in the swimbladder of individual eel specimens can be monitored easily.     

Changes in otolith microchemistry of the Japanese eel, Anguitta japonica, during its migration from the ocean to the rivers of Taiwan

The newly recruited Japanese eel, Anguilla japonica , elvers and 1-year-old eels collected in estuaries and in rivers, respectively, were studied. The microstructure and chemical composition of the sagittal otolith of these eels were examined by SEM and wavelength-dispersive spectrometer (WDS), A transition zone or'elver mark'was observed in the otolith of the young eels. A comparison of the otoliths of elvers with those from the 1-year-old eels suggests that this transition zone was deposited during upstream migration, a change from a marine to freshwater environment. Strontium (Sr) content in the primordium of the otolith of both elvers and young eels was low, probably due to the maternal or freshwater origin of the oocyte. The concentration of Sr in the otolith increased gradually during marine life and reached a peak approximately 1 month before upstream migration. As the elvers entered the estuary, the Src concentration dramatically decreased and remained at a low level thereafter. These findings indicate that the history of the migratory environment of the eel can be reconstructed from a combined study of otolith microstructure and microchemistry analysis.     

Pharmacokinetics of ciprofloxacin in eels by high-performance liquid chromatography with fluorescence detection

The plasma kinetics of ciprofloxacin (CF) were investigated in the eels after administration by oral gavage and bath treatment. Plasma concentrations of CF were determined by high-performance liquid chromatography with fluorescence detection. The mean concentration time data after oral gavage of a single dose (10.0 mg/kg CF) and after bath treatment by exposure (10 microg/ml CF) to medicated water for 48 h were both best fitted by a one-compartment model. After oral gavage in eels, the half-time of absorption (T1/2Ka) was 0.10 h, the half-time of elimination (T1/2Ke) was 51.87 h, and the maximum plasma concentration (Cmax) was 0.4552 microg/ml at Tmax 0.88 h. After bath treatment, the (T1/2Ka) was 0.02 h, the (T1/2Ke) was 15.46 h, and the Cmax was 0.1175 microg/mL at Tmax 0.22 h.     

Riparian land use and accessibility to fishers influence size class composition and habitat use by longfin eels in a New Zealand river

Fyke nets were used to sample longfin eels Anguilla die.enbachii in fourth order stream sites with contrasting riparian land use (native tussock, exotic pasture and pasture plus willows Salix spp.) in Lee Stream, a tributary of the Taieri River, New Zealand. Total lengths (L_T) of longfin eels from locations inaccessible to eel fishers were normally distributed whereas those from accessible locations had a non-normal, positively skewed distribution. Mean L_T and body condition were higher in inaccessible than accessible sites, consistent with considerable fishing exploitation in the latter. Mean L_T of the fish was greatest in pasture, intermediate in willow and smallest in tussock sites. Larger longfin eels (≥535 mm) were associated with a riffle-type habitat consisting of shallow, faster-flowing water with coarse, variable sediment and were taken farther from cover. In contrast, smaller longfin eels were associated with a pool-type habitat consisting of slower-flowing, deeper water, with fine, homogenous sediment and were captured closer to cover. This pattern of habitat use differed from some previous reports and probably reflects differences in methodology: fyke nets set over 48 h to sample actively moving longfin eels compared to daytime electric fishing, which samples longfin eels that are more likely to be at rest.