Estimation of gonadal status of Japanese female eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> by a single injection of salmon pituitary extract

ABSTRACT:   Cultivated eels often either lack a part of the ovary (abnormal) or do not have any gonadal tissue (completely sterile). These abnormalities may cause a failure of induction of sexual maturation of female eels. In order to effectively induce sexual maturation and obtain good quality eggs, female eels having normal ovaries should be selected before induction of maturation. The present study examined effects of injection of salmon pituitary extract (SPE) on serum estradiol-17β levels to estimate gonadal status of sexually immature Japanese eels. At the initial stage (just prior to SPE injection), serum estradiol-17β levels were low and no significant difference was observed among eels possessing the different gonad types (normal, abnormal, and sterile), suggesting that gonadal status cannot be estimated by the measurement of serum estradiol-17β levels. A single injection of SPE significantly increased serum estradiol-17β levels in eels possessing normal and abnormal ovaries, but not in sterile eels. Serum estradiol-17β levels significantly increased from 24 h and were high on day 3. These data indicate that eels possessing ovaries can be distinguished from sterile eels by measurement of serum estradiol-17β between 1 and 3 days after a single injection of SPE.     

Effects of silvering state on induced maturation and spawning in wild female Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

ABSTRACT:   The effects of silvering state of wild female Japanese eels Anguilla japonica on the success of induced maturation and the following spawning were examined. Thirty-eight females, collected in Mikawa Bay, were divided into four stages based on their silvering state: yellow (Y1), late-yellow (Y2), silver (S1) and late silver eels (S2). Despite injections of salmon pituitary extract (SPE) through the standard technique, Y1 and Y2 eels did not respond to the treatment with undeveloped gonad (gonad-somatic index [GSI]: 0.3–0.9), and all these females died by 5 weeks, probably due to an abnormal physiological condition. Most S1 (81%) and S2 eels (100%) matured completely (GSI: 17.8–51.4), and finally spawned successfully (69% for S1, 89% for S2). S2 eels fully matured with oocytes of over 750 μm in diameter by significantly smaller number of injections of SPE (5–6 times) than the case of S1 eels (6–8 times). The amount of eggs released by S2 eels (0.65 ± 0.11 g/fish per body weight [BW]) was significantly larger than those by S1 eels (0.54 ± 0.09 g/fish per BW). There was no difference in fertilization and hatching rates between eggs released by S1 eels and those of S2 eels. These results indicate that the success of induced maturation and spawning in wild female Japanese eels depends on their silvering state, and matured eggs can be obtained efficiently through the use of S2 eels rather than other stages.     

Lipid content and fatty acid composition of muscle,liver, ovary and eggs of captive-reared and wild silver Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> during artificial maturation

ABSTRACT:   Changes in lipid content and fatty acid composition of muscle, liver and ovary of captive-reared and wild silver Japanese eel Anguilla japonica were examined during artificial maturation induced by salmon pituitary homogenate (SPH) injections. Although the relative levels of n-3 and n-6 highly unsaturated fatty acids (HUFA) in liver and ovary were higher than in muscle in both captive and wild silver eels before SPH injection, these tended to decrease with maturation. The relative levels of n-6 HUFA in muscle, liver, ovary and eggs of wild silver eels were remarkably higher than those in captive eels. Therefore, we attempted to alter the ratio of n-6 HUFA in eggs by feeding eels a diet supplemented with linoleic acid-rich plant oil. Although the percentage of n-6 polyunsaturated fatty acids to total fatty acids in eggs of eels fed the supplemented diet was similar to wild silver eels, the percentage of n-6 HUFA remained remarkably lower than in wild silver eels. Hence, it appears that the supplemented diet affected the fatty acid composition of eggs, but did not result in much conversion of linoleic acid to its higher homologs in eels.     

Isolation of a novel polyomavirus,related to Japanese eel endothelial cell‐infecting virus,from marbled eels,Anguilla marmorata (Quoy & Gaimard)

Marbled eels, Anguilla marmorata (Quoy & Gaimard), cultured in Taiwan exhibited haemorrhage and mortality in January 2012. The severely diseased eels bled from the gills and showed congestion of the central venous sinus of the gill filaments and haemorrhage throughout the body similar to viral endothelial cell necrosis of eel. In this study, a novel polyomavirus (AmPyV) was isolated from the diseased eels using the AMPF cell line established from the pectoral fin of healthy marbled eels. AmPyV was found to encode a long T‐antigen orthologous gene. Phylogenetic analysis showed that AmPyV was closely related to Japanese eel endothelial cell‐infecting virus. PCR assays revealed AmPyV infection throughout the systemic organs. AmPyV proliferated in the AMPF, EK‐1 and EO‐2 cells at temperatures 25–30 °C, and the progeny virus yields were 10^7.0, 10^7.4 and 10^7.7 TCID₅₀ mL^?1, respectively. The purified virions were icosahedral particles, 70–80 nm in diameter. No clinical signs or mortality was observed among the eels injected with the virus; however, the virus was reisolated from the brain, eyes, kidneys, fins and gills of infected eels 2 month after injection. Our results suggest that AmPyV exhibits a latent infection. Pathogen of the disease needs to study further.     

Immunohistochemical changes in production of pituitary hormones during artificial maturation of female Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

ABSTRACT:   Specific antibodies against follicle-stimulating hormone β subunit (FSHβ), prolactin (PRL), and somatolactin (SL) of the Japanese eel Anguilla japonica were produced. These antibodies, as well as antibodies against luteinizing hormone β subunit (LHβ) and growth hormone (GH) produced previously, were used to examine changes in the production of pituitary hormones in female eels during maturation induced by salmon pituitary homogenate (SPH) injection. Immunohistochemical observations showed a decrease in FSH production after SPH injection, suggesting that SPH inhibits FSH production. In contrast, LH production increased markedly with maturation. The number of GH producing cells decreased gradually during maturation, possibly because of inhibition by exogenous GH present in the SPH and/or endogenous insulin-like growth factor-I produced by the stimulation of salmon GH. Although changes in the number of PRL producing cells with maturation were not evident, the number of SL producing cells showed a peak at the late vitellogenic stages, and thereafter decreased to the migratory nucleus stage. These results suggest that GH and SL are involved in sexual maturation in SPH injected eels, as in other fishes.     

Growth differences of Japanese eels Anguilla japonica between fresh and brackish water habitats in relation to annual food consumption in the Kojima Bay‐Asahi River system,Japan

To learn about the relationships between feeding and growth of temperate eels in freshwater and brackish water habitats, we analysed 533 yellow‐phase Japanese eels Anguilla japonica collected in both types of habitats in southeastern Japan. Because male eels were very rare in each habitat (FW, N = 1; BW, N = 20), characteristics of female eels were compared between the different habitats. Annual food consumption was evaluated with the consideration of instantaneous food consumption and annual activity period. Stomach fullness index (stomach content weight/body weight) was used as an indicator of instantaneous food consumption. The ratios of number of months with eel catch to those when eel sampling was conducted were used as an indicator of activity period. Female yellow eels tended to be older and slower growing in fresh water (N = 78; age, mean ± SD = 7.9 ± 2.4 years; growth rate, 59.8 ± 14.0 mm year^?1) than in brackish water (N = 229; age, 5.5 ± 1.8 years; growth rate, 90.1 ± 24.4 mm year^?1). Irrespective of sex, yellow eels in brackish water had a higher stomach fullness index and a greater ratio of months with eel catches, indicating greater annual food consumption by brackish water eels. These results indicate that greater annual food consumption contributes to the greater growth rates of Japanese eels in brackish water habitats.     

Two forms of cytochrome P450 cDNA from 3-methylcholanthrene-treated European eel Anguilla anguilla

ABSTRACT:     The cytochrome P450 (CYP) represents a large group of microsomal monooxygenases that catalyze drugs as well as a host of lethal environmental contaminants such as dioxins, leading to either detoxification and excretion from the animal or generation of carcinogenic intermediates. In the present study two forms of cDNA were cloned (Eu MC1 and Eu MC2) for European eel CYP1A genes by polymerase chain reaction (PCR) techniques. The cDNA of Eu MC1 was 3368 bp long coding 521 amino acid residues, and that of Eu MC2 was 2464 bp long coding 517 amino acid residues. Identities of deduced amino acid sequences between Eu MC1 and Japanese eel CYP1A1 and that between Eu MC2 and the second form of Japanese eel CYP1A were 98% and 97%, respectively, showing decisively that Eu MC1 and Eu MC2 are orthologous to Japanese eel CYP1A1 and the second form of CYP1A, respectively. A striking difference between the two eel species was that the Eu MC1 peptide was two amino acid residues longer than that of the Japanese eel CYP1A1. Existence of two loci of CYP1A in Japanese and European eels may suggest that the two forms of CYP1A exist widely among the eel species, because the divergence between the two eel species has been shown to be close to the basal divergence among eels. The identities in CYP1A may help to estimate genetic distance between European and Japanese eels.     

Effects of temperature on vitellogenesis in Japanese eel Anguilla japonica

ABSTRACT:   Cultured immature female Japanese eels acclimated to sea water at either 10 or 20°C were treated weekly with salmon gonadotropin fraction (sGTH) in order to investigate the effects of water temperature on artificial induction of ovarian maturation. In eels maintained at 20°C, ovulation was induced in 11 of 18 fish during the experimental period of 13 weeks, whereas at 10°C all fish showed a low gonadosomatic index (GSI) at the end of the experiment. Plasma vitellogenin levels were higher in eels kept at 20°C than in eels at 10°C throughout the experiment. However, no significant differences were observed in the plasma testosterone and estradiol-17β levels between groups. Eels pretreated with sGTHs at 10°C for 13 weeks were separated to two different temperature (10 and 20°C) groups, and received the same weekly sGTH injections. In eels transferred to 20°C, plasma vitellogenin levels, GSI and oocyte diameter were increased, but these values were maintained at low levels in eels that remained at 10°C. These results clearly indicate that water temperature is an important factor regulating vitellogenesis in the Japanese eel.     

Antibody production in Japanese eels, Anguilla japonica Temminck & Schlegel

Adult Japanese eels, Anguilla japonica Temminck & Schlegel, (200–250 g, 45–55 cm) were immunized by intramuscular injection with goat IgG. After 5 weeks, eel immunoglobulin (Ig) was purified using affinity chromatography. The purified eel Ig was used to immunize rabbits to produce anti-eel Ig antibody. The highest antibody ELISA value in eels was reached 3 weeks after initial immunization with goat IgG, and then gradually decreased. The antibody could still be detected at 140 days post-immunization. The optimal temperature for antibody production was 30°C. Freund's complete adjuvant and secondary immunization both increased antibody production in eels.     

Influence of geomagnetism on the activity and orientation of the eel, Anguilla anguilla (L.), as evident from laboratory experiments

Abstract– Studies were performed with both a mechanical and a photosensory arrangement of the experimental tanks. Silver (migratory) eels were more active than yellow (stationary) eels. Comparison of pooled data on yellow eels tested under natural geomagnetic field conditions with those on eel tested under unnatural ones prcded no evidence of difference in activity. Silver and yellow eels showed stronger activity during the 6 days around new moon compared with the phase around full moon as well as a peak before new moon and a smaller peak before full moon. Under each kind of unnatural field condition, the activity relationships reversed or were cancelled. The directional choice of eels under natural conditions (controls) pointed to magnetic NW and mostly also in the opposite direction. When the magnetic north was changed by 90°, the tested yellow eels followed this change to about the same degree. The additional change of both the inclination (from 68° or 45° or 30°) and total intensity (by a reduction to 31% of the natural field) resulted in a change of the preferences of about 90° relative to magnetic north. Reversal of magnetic north by 180°, which is identical to a change of the inclination from 68° to 112°, also resulted in a change of the preferences by about 90°. Three experiments using compensation of both the horizontal and the vertical geomagnetic field led to circular directional preferences at random in two cases and in one case to about NW of the residual field magnetic north. The navigational abilities of the eel, on the basis of its magnetic sensitivity, are discussed.     

Oogenesis in the common Japanese conger Conger myriaster

ABSTRACT:    We investigated the process and characteristics of oogenesis in the common Japanese conger Conger myriaster . Young fish caught in November 1996 were reared for use in this experiment. Fish were sampled monthly from December 1997 to August 1998. Some were injected with human chorionic gonadotropin to stimulate ovarian maturation from May to August 1998. Oocytes from the chromatin nucleolus stage to the secondary yolk globule stage were obtained from non-hormone-treated fish; those of more advanced stages were obtained from hormone-treated ones. We divided oocyte development into eight stages from the chromatin nucleolus stage to the maturation stage. The yolk vesicle stage was not separated because yolk vesicles began to appear just after appearance of yolk globules. Oocyte, oil droplet, yolk globule and nucleus diameters all increased concomitant with oocyte development. Oil droplet and yolk globule diameters increased remarkably at the maturation stage. However, zona radiata thickness peaked at the secondary yolk globule stage, decreasing gradually thereafter. Increased gonadosomatic index was related to oocyte development as found in European and Japanese eels receiving hormone treatment to mature. The present study is the first report describing oogenesis characteristics in congrid eels. It indicates that oogenesis is almost identical to that of other anguillid eels.     

Environmental DNA analysis reveals the spatial distribution,abundance, and biomass of Japanese eels at the river‐basin scale

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Immune effects of bathing European eels in live pathogenic bacteria,Aeromonas hydrophila

The specific and non‐specific immune parameters and protection of European eels (Anguilla anguilla) were evaluated after bathing eels with Aeromonas hydrophila. Two hundred eels were distributed into two equal groups and bathed with Phosphate‐buffered saline (Control group) or 1.0 × 10⁷ cfu mL^?1 A. hydrophila (Test group) for 1 h respectively. Then, eels were bled aseptically from the caudal sinus on 1, 4, 7, 14 and 28 days post treatment. The blood cells were used to evaluate the cellular immunity and the serum was used to determine the titres of specific antibody as well as the activities of superoxide dismutase (SOD) and lysozyme. Eels from both groups were challenged by intraperitoneal injection of 1.0 × 10⁷ cfu of A. hydrophila on 28 and 42 days post bathing. The results show that eels bathed in A. hydrophila significantly (P < 0.05) enhanced the proliferation of different types of blood cells and the serum titres of anti‐A. hydrophila antibody. The Relative Percent Survival (RPS) after challenge on 28 and 42 days post bathing in Test group vs. Control group was 40% and 50% respectively. These results suggest that bathing European eels in A. hydrophila would positively affect specific as well as non‐specific immune parameters and protect against A. hydrophila infection in freshwater farming.     

Variation in the demographic characteristics of yellow‐phase Japanese eels in different habitats of the Hamana Lake system,Japan

Abstract – Yellow‐phase Japanese eels (Anguilla japonica) were investigated in the Hamana Lake system, Japan, from 2003 to 2004 to understand how their demographic attributes vary within the lake system. A total of 779 yellow eels were collected during sampling in two inlet rivers and two brackish/saltwater lakes within the lake system. Female eels predominated, constituting 84% of the 75 sex‐determined eels in the river, and 50% of the 151 sex‐determined eels in the lakes. Total lengths (TL) of all eels examined ranged from 54.2 to 715.0 mm (mean = 320.4 ± 145.4 SD). In the inlet river, the TL of eels showed a significant positive relation with the distance from the river mouth. The estimated relative abundances of eels ranged from 0 to 1.8 eels·m^?2 effort (mean: 0.3 ± 0.41) in the river and was negatively correlated with the distance from the river mouth. This suggested that larger eels might tend to be distributed at lower abundances in upstream reaches of the river. Mean age of yellow eels determined by their otolith annuli was younger in the lake (N = 117, 3.3 ± 1.4 years) than in the river (N = 214, 4.3 ± 1.7 years). Growth rate was higher in the lake than in the river at age 1–2 years (131.9 and 104.4 mm·year^?1, respectively). The results of this study suggest that, although Japanese eels can adapt to various types of environments, significant differences can occur in population structures and growth patterns among habitats.     

A preliminary telemetry study of the migration of silver European eel (Anguilla anguilla L.) in the River Mosel, Germany

Abstract  – To study the behaviour of silver eels ( Anguilla anguilla L.) during their downstream migration, particularly near a hydroelectric power dam, we tagged nine eels with ultrasonic transmitters and tracked their paths in the River Mosel, Germany. The onset of migration coincided with the first flood event that followed the full moon but was independent of daytime, because migration and turbine passage occurred during both day and night. During migration eels swam actively downstream with a velocity of 0.3–1.2 m · s⁻¹. When migrating eels arrived at the dam, they either passed through the turbines immediately or stayed upstream of the powerhouse for up to 8 days, showing a characteristic circling behaviour. Circling eels repeatedly approached the trashrack, sprinted upstream, and finally passed through the turbines with the next high water discharge. These observations are discussed with regard to the design of appropriate downstream passage facilities.     

Discovering the dominance of the non-native European eel in the upper reaches of the Tone River system,Japan

To investigate the presence of non-native anguillid eels in Japanese waters, 141 eels were collected from seven sampling sites throughout the Tone River system. Genetic species identification showed an extraordinary dominance of the non-native European eel Anguilla anguilla in the uppermost site of the study area. Estimation of age from otoliths suggested that the European eels were introduced into the river in the 1990s, corresponding to previous reports from other Japanese water systems. Comparison of the von Bertalanffy growth curve parameters indicated that the European eels caught in the Tone River system appeared to have a similar or even higher growth rate than the same species in the original habitats in Europe. The long-term inhabitation and the normal development of European eels in Japanese waters suggest that regulations prohibiting the release of non-native eels and safeguards against accidental escape from culture ponds must be strictly maintained in order to ensure the conservation of the native Japanese eel.     

Comparative study of Japanese eel and yellowfin goby brains using a new brain size index

We propose a simple index of brain volume (D*L index), calculated using two-dimensional photographs of the brain, captured from the dorsal and lateral views. Here, D and L represent the dorsal and lateral brain areas in the images, respectively. Two distantly related species were assessed. The D*L indices of 11 Japanese eels, Anguilla japonica and 11 yellowfin gobies, Acanthogobius flavimanus were calculated. High correlation coefficients of D*L index with brain weights were obtained from both species, suggesting the validity and reliability of the index. The relative profiles of the olfactory bulb, telencephalon, optic tectum, and cerebellum were calculated using this index. Eels had a larger olfactory bulb and telencephalon, whereas gobies had a larger optic tectum. Principal component analysis showed two separate clusters for eels and gobies. Published images of two Anguilliformes species and four Gobiidae species were used to test the D*L index. Principal component analysis showed separate clusters for Anguilliformes and Gobiidae species. The D*L index is a suitable tool for data collection because it is simple, low-cost, and has reliability and validity.

     

Development and validation of a real‐time PCR assay for the detection of anguillid herpesvirus 1

Anguillid herpesvirus 1 (AngHV1) causes a haemorrhagic disease with increased mortality in wild and farmed European eel, Anguilla anguilla (L.) and Japanese eel Anguilla japonica, Temminck & Schlegel). Detection of AngHV1 is currently based on virus isolation in cell culture, antibody‐based typing assays or conventional PCR. We developed, optimized and concisely validated a diagnostic TaqMan probe based real‐time PCR assay for the detection of AngHV1. The primers and probe target AngHV1 open reading frame 57, encoding the capsid protease and scaffold protein. Compared to conventional PCR, the developed real‐time PCR is faster, less labour‐intensive and has a reduced risk of cross‐contamination. The real‐time PCR assay was shown to be analytically sensitive and specific and has a high repeatability, efficiency and r²‐value. The diagnostic performance of the assay was determined by testing 10% w/v organ suspensions and virus cultures from wild and farmed European eels from the Netherlands by conventional and real‐time PCR. The developed real‐time PCR assay is a useful tool for the rapid and sensitive detection of AngHV1 in 10% w/v organ suspensions from wild and farmed European eels.     

日本鳗鲡产卵后生存及其繁育

为证明日本鳗鲡(Anguilla japonica)生活史最后一步一产后鳗的命运,本研究模拟产后的日本鳗鲡继续在海水中养殖,观察其存活率及繁育情况.结果表明,产后鳗在海水中停食约18 d后,体能得到恢复,部分亲鱼开始出现摄食,1个月左右全部恢复摄食,经244 d养殖,雌、雄鳗体质量增加,存活率达94.6%.随后,给产后鳗注射外源性促性腺激素(鲤鱼脑垂体匀浆CPE和人绒毛膜促性腺激素HCG)后激发其退化的性腺(卵巢和精巢)重新发育(与当年银鳗作对照).通过性腺组织切片观察产后鳗和对照鳗性腺发育成熟的全过程及其差异,发现产后鳗起初性腺发育比当年银鳗差,但经多次注射激素后,产后鳗性腺成熟与当年银鳗同步,证明产后鳗生殖细胞对激素的敏感件高.应用17α,20β-双羟孕酮和促性腺激素释放激素类似物(GnRH-A3)使催熟的产后鳗和对照鳗均产卵和排精,并孵化出仔鱼,从而有力地证明,鳗鲡产后虽体质弱,但待体能恢复后能够继续生存和繁殖.本研究旨在探讨利用产后鳗作为今后鳗鲡人工繁殖亲鳗的可行性.