Development of a novel Japanese eel myoblast cell line for application in cultured meat production.

The present study investigates the isolation, analysis, and characterization of primary cultured cells derived from the muscle tissue of Japanese eel (Anguilla japonica), culminating in establishing a spontaneously immortalized myoblast cell line, JEM1129. We isolated satellite cells from eel muscle tissue to establish a foundation for cultured eel meat production. While initial cell cultures contained myoblasts, continued passaging led to a decline in myoblast characteristics and an increase in fibroblast-like cells. RNA-Seq and RT-qPCR analyses showed significant downregulation of well-established markers for satellite cells and myoblasts, such as pax7a and myoD, over successive passages, highlighting a loss of myoblastic traits. Single-cell cloning was employed to overcome this challenge and maintain myoblast purity, leading to the successful creation of the JEM1129 cell line. These JEM1129 cells demonstrated enhanced expression of myoblast marker genes, exceeding the initial primary culture cell population. The cells showed strong myotube formation, particularly when cultured in a differentiation medium, indicating their robust potential for muscle development. The JEM1129 cell line represents a significant advancement in the cultivation of eel muscle cells, offering a promising avenue for cultured meat production. The findings contribute to a deeper understanding of muscle cell biology and provide valuable insights into using fish-derived myoblasts for cultured meat production.

Mathematical lipid correction of δ13C and effect of lipid extraction on δ15N of European eel (Anguilla anguilla) muscle: Lipid correction of δ13C in European eels.

RATIONALE: Carbon (δ13C) and nitrogen (δ15N) stable isotope analysis is a powerful tool to investigate diverse questions in fish ecology, such as their trophic position or migration strategies. These questions appear particularly important to protect endangered European eel. However, elevated lipid content in eel muscle can bias δ13C values, as lipids are 13C-depleted compared to proteins and carbohydrates. METHODS: We measured δ13C and δ15N values of bulk and lipid-free samples of eel muscle. Lipid-free samples were obtained after the extraction of lipids with cyclohexane. Lipid-corrected δ13C values, using five different mathematical equations based on bulk δ13C values, were compared to lipid-free δ13C values. We also evaluated the effect of lipid extraction on δ15N values. The analyses were based on linear regression performed on 333 individuals captured in nine lagoons and four rivers. RESULTS: Independently to the capture site or habitat (river or lagoon), the predicted lipid-corrected δ13C values were highly consistent with the measured lipid-free δ13C values (R2 > 0.90). The application of specific equations for each habitat or capture site only slightly increases these R2 (1.5% or less). The lipid extraction treatment significantly decreased by 0.2‰ the δ15N values compared to bulk samples. CONCLUSIONS: Given the excellent prediction of mathematical equations and the small decrease of δ15N values after lipids extraction, we propose to use mathematical correction to estimate δ13C values of eel muscle. As the habitats or sites did not strongly influence the results, the coefficients from our study can be applied to other studies on European eel.

How Japanese eels escape from the stomach of a predatory fish.

Predation shapes diversity in the defensive tactics of prey. One specialized defensive tactic is to escape the digestive system of the predator after capture1,2,3,4,5,6,7,8. While most of these defensive tactics involve passive ejection alive from predators' mouths and vents1,2,3,4,5, active escape from the digestive tracts of predators has recently been observed in certain invertebrate species6,7 and fish8. However, no study has yet uncovered the behavioral patterns and escape routes of the prey within a predator's digestive tract. Here, we report the sequential escape processes of the Japanese eel Anguilla japonica from capture to escape via the gills of predatory fish Odontobutis obscura using an X-ray video system. All captured eels had at least one portion of their bodies swallowed into the stomach of the predator. Surprisingly, after being swallowed, most individuals attempted to escape by going back up the digestive tract towards the esophagus and gill, and some of them succeeded in escaping via the predator's gill. Some eels, whose entire bodies were completely inside the stomach, exhibited circling behavior along the stomach, seemingly searching for possible escape routes. An electro-anesthetization experiment revealed that eels utilize various escape routes through gill clefts, rather than just one.

Laying the foundations for selective-fish guidance using electricity: multi-species response to pulsed direct currents.

To develop effective technology that employs electric fields to simultaneously guide valued freshwater fish whilst limiting the range expansion of undesirable invasive species, there is a need to quantify the electrosensitivity of multiple families. This experimental study quantified the electrosensitivity of two carp species that, in UK, are invasive (grass carp, Ctenopharyngodon idella, and common carp, Cyprinus carpio) and compared the values with those previously obtained for adult European eel (Anguilla anguilla), a species of conservation concern in Europe. Electric field strengths (V/cm) required to elicit physiological responses (twitch, loss of orientation and tetany) were identified across four pulsed direct current (PDC) electric waveforms (single pulse-2 Hz, double pulse-2 Hz, single pulse-3 Hz and double pulse-3 Hz). Grass carp were sensitive to differences in waveform with tetany exhibited at lower field strengths in the single pulse-2 Hz treatment. Both cyprinid species responded similarly and were less sensitive to PDC than adult European eel, although loss of orientation occurred at lower field strengths for grass than common carp in the single pulse-3 Hz waveform treatment. This variation in electrosensitivity, likely due to differences in body length, indicates potential for electric fields to selectively guide fish in areas where invasive and native species occur in sympatry.

River-estuary continuum highlighted by variabilities in carbon and nitrogen stable isotope ratios of the catadromous eel Anguilla japonica.

Estuaries exhibit high physicochemical variability and the properties of estuaries and the constituent segments are not yet systematically understood. This study aims to reveal the spatial heterogeneity of predominant organic sources using carbon and nitrogen stable isotope ratios (δ13C and δ15N, respectively) of Japanese eels (Anguilla japonica), one of the ideal natural samplers. In the Miyakoda River, Japan, our study site, the effectiveness of eel δ13C and δ15N values as standard indices of predominant organic sources was tested by employing the River Continuum Concept. This study then extended the application of these indices to the estuary, i.e., the Hamana Lake system, into which the Miyakoda River flows. Once in the upper estuary, eel δ13C and δ15N values became high, with the latter peaking in this river-estuary continuum, indicating that artificial labile subsidies (nutrients and organic matter) with high δ15N values were rapidly assimilated into the food web. Eel δ15N values decreased again in the middle estuary. Nevertheless, the influence of terrestrial organic subsidies extended into this segment, as evidenced by the low δ13C values of eels. These results suggest that refractory organic matter with low δ15N values, such as plant-derived ones, is slowly assimilated into the food web in the downstream estuarine segments. The higher δ13C values in the lower estuary suggested that the contribution of eelgrass or macroalgae occurred in addition to benthic microalgae. Thus, our results emphasize the need to consider the multiple energy flows to understand the estuary as a continuum.

Highly contiguous genome assembly and gene annotation of the short-finned eel (Anguilla bicolor pacifica).

In East Asia, anguillid eels are commercially important. However, unlike other species, they have not been successfully cultivated throughout their lifecycle. Facing population decline due to overharvesting and environmental pressures, the industry is turning to alternatives, such as Anguilla bicolor pacifica (short-finned eel). However, genomic data for short-finned eels are unavailable. Here, we present in-depth whole-genome sequencing results for short-finned eel obtained using two sequencing platforms (PacBio Revio, and Illumina). In this study, we achieved a highly contiguous genome assembly of the short-finned eel, comprising 19 pseudochromosomes encompassing 99.76% of the 1.087 Gb genome sequence with an N50 of 16.88 and 61.07 Mb from contig and scaffold, respectively. Transcripts from four different tissues led to the annotation of 23,095 protein-coding genes in the eel genome, 98.66% of which were functionally annotated. This high-quality genome assembly, along with the annotation data, provides a foundation for future functional genomic studies of short-finned eels.

Evidence of striking morphological similarity and tissue tropism of phylogenetically distant myxozoan genera: Myxidium and Paramyxidium in the kidney of the European eel.

European eel, Anguilla anguilla (Linnaeus) (Elopomorpha: Anguilliformes), is a critically endangered fish of ecological and economic importance, hosting numerous parasites, including myxozoans (Cnidaria). Since its initial discovery in the kidney of European eel, Myxidium giardi Cépède, 1906 has been reported with numerous spore sizes and shapes from various tissues of multiple anguillid species. Morphological variability, wide host and tissue spectrum, and lack of sequence data raised doubts about the conspecificity of reported isolates. Subsequent studies provided 18S rDNA sequences of several isolates from anguillids and other elopiform fish, and demonstrated a split of parasite data into two distinct phylogenetic lineages, one comprising the M. giardi sequence, and the other all species infecting elopiform fishes classified under the recently established genus Paramyxidium Freeman et Kristmundsson, 2018. Myxidium giardi was, however, transferred to this genus as Paramyxidium giardi n. comb. and designated as the type species of the genus. In line with this change, the sequence originally identified as M. giardi was considered to have been incorrectly associated with this species. To shed light on the status of M. giardi originally described by Cépède (1906), we conducted microscopic and molecular examinations of various organs of 24 individuals of European eel, originating from diverse Czech habitats. Through morphometric and molecular analyses, we demonstrated that spore and polar capsule morphology, morphometry and tissue tropism of our European eel kidney parasite isolates matched the features of the original M. giardi description. Our isolates clustered in the lineage encompassing the first published M. giardi sequence. Thus, the originally described M. giardi indeed represents an existing species within the genus Myxidium Bütschli, 1882, which we formally resurrect and redescribe. Due to the morphological and molecular differences between M. giardi and P. giardi of Freeman et Kristmundsson (2018), we additionally rename the latter species as Paramyxidium freemani nom. nov.

Rethinking fish-friendliness of pumps by shifting focus to both safe and timely fish passage for effective conservation.

Globally, catadromous freshwater eels of the genus Anguilla are of conservation concern, including critically endangered European eel (Anguilla anguilla). Pumping stations that move river water to a higher elevation severely impact eels during their seaward spawning migration. Fish-friendly pumps can mitigate fish injury and mortality but here we uniquely rethink a fish-friendly pump as a fish passage solution. In this pluriannual study, the seasonal timing of pump operation was misaligned with the typical silver eel migration period. Eels were almost exclusively nocturnal but night-time pumping represented as little as 5.6% a year. Night-time eel approaches were primarily influenced by pump duration and temperature, but did not align with lunar phase, unlike in unregulated rivers. After reaching the pumping station, eel passage was influenced by weedscreen aperture and increased when the aperture was increased. Passive sensor collision suggested non-pump infrastructure could cause injury and mortality to eels. It is therefore recommended pump operation should align with the timing of silver eel migration, weedscreen and pump entrance efficiencies should be maximised, and non-pump infrastructure must have low fish injury risk. Ultimately, considering the entire structure a fish passage solution will help ensure fish-friendly pumps have high conservation value for anguillid eels globally.

Environmental DNA Reveals Geographic Distributions of Two eel Species, Anguilla japonica and A. marmorata, in Western Kyushu, Japan.

Some anguillid eels migrate thousands of kilometers from their spawning grounds, dispersing across vast geographic areas to fresh and brackish water habitats, where they settle and grow. Japanese eels (Anguilla japonica) and giant mottled eels (A. marmorata) are both found in Japan, although their distributions differ, and their exact distributions are poorly known. We assumed that topographic distribution patterns of Japanese and giant mottled eels must differ among and within rivers along the northwest coast of Kyushu, Japan. Environmental DNA (eDNA) analysis was conducted at 87 sites in 23 rivers. Japanese eel eDNA was detected in 19 rivers (82.6%) and that of giant mottled eels was detected in eight (34.8%). We detected giant mottled eel eDNA in five rivers where they were previously unknown. eDNA for Japanese eels was detected at six of nine sites in the north (66.7%), 13 of 23 sites in Omura (56.5%), and 37 of 55 sites in the south (67.3%). In contrast, giant mottled eel eDNA was detected at one of nine sites in the north (11.1%), no sites in Omura, and 15 of 55 sites in the south (27.3%). There was no correlation between eDNA concentrations of the two species at 10 sites in the five rivers where eDNA of both species was detected. These findings suggest differences in the distribution of the two eel species and the northern distributional limit of giant mottled eels in the area facing the East China Sea.

Microbial influence on the larval survival of Japanese eel Anguilla japonica: Antibiotic-mediated alterations and biomarker isolation.

In rearing systems for the Japanese eel Anguilla japonica, although it is assumed that microorganisms influence larval survival and mortality, particularly during the early stages of growth, the effects of bacterial communities on larval survival have yet to be sufficiently determined. In this study, we compared the bacterial communities associated with larval survival at three stages of eel growth. To artificially alter bacterial communities and assess larval survival, eel larvae were treated with 11 types of antibiotic, and larval survival and bacterial characteristics were compared between the antibiotic-treated and antibiotic-free control groups. Throughout the three growth stages, eels treated with four antibiotics (polymyxin B, tetracycline, novobiocin, and erythromycin) had survival rates higher than those in the control groups. The bacterial communities of surviving larvae in the control and antibiotic groups and dead larvae in the control groups were subsequently analyzed using 16S rRNA gene amplicon sequencing. PERMANOVA analysis indicated that these three larval groups were characterized by significantly different bacterial communities. We identified 14 biomarker amplicon sequence variants (ASVs) of bacterial genera such as Oceanobacter, Alcanivorax, Marinobacter, Roseibium, and Sneathiella that were enriched in surviving larvae in the antibiotic treatment groups. In contrast, all four biomarker ASVs enriched in dead larvae of the control groups were from bacteria in the genus Vibrio. Moreover, 52 bacterial strains corresponding to nine biomarkers were isolated using a culture method. To the best of our knowledge, this is the first study to evaluate the bacterial communities associated with the survival and mortality of larvae in during the early stages of Japanese eel growth and to isolate biomarker bacterial strains. These findings will provide valuable insights for enhancing larval survival in the eel larval rearing systems from a microbiological perspective.

Deep antiquity of seagrasses supporting European eel fisheries in the western Baltic.

Protecting ocean habitats is critical for international efforts to mitigate climate impacts and ensure food security, but the ecological data upon which policy makers base conservation and restoration targets often reflect ecosystems that have already been deeply impacted by anthropogenic change. The archaeological record is a biomolecular archive offering a temporal scope that cannot be gathered from historical records or contemporary fieldwork. Insights from biogeochemical and osteometric analyses of fish bones, combined with context from contemporary field studies, show how prehistoric fisheries in the western Baltic relied on seagrass meadows. European eels (Anguilla anguilla) harvested by Mesolithic and Neolithic peoples over millennia showed a strong fidelity for eelgrass foraging habitats, an ecological relationship that remains largely overlooked today, demonstrating the value of protecting these habitats. These data open new windows onto ecosystem- and species-level behaviours, highlighting the need for wider incorporation of archaeological data in strategies for protecting our oceans.

Transcriptome analysis of the Japanese eel (Anguilla japonica) during larval metamorphosis.

BACKGROUND: Anguillid eels spend their larval period as leptocephalus larvae that have a unique and specialized body form with leaf-like and transparent features, and they undergo drastic metamorphosis to juvenile glass eels. Less is known about the transition of leptocephali to the glass eel stage, because it is difficult to catch the metamorphosing larvae in the open ocean. However, recent advances in rearing techniques for the Japanese eel have made it possible to study the larval metamorphosis of anguillid eels. In the present study, we investigated the dynamics of gene expression during the metamorphosis of Japanese eel leptocephali using RNA sequencing. RESULTS: During metamorphosis, Japanese eels were classified into 7 developmental stages according to their morphological characteristics, and RNA sequencing was used to collect gene expression data from each stage. A total of 354.8 million clean reads were generated from the body and 365.5 million from the head, after the processing of raw reads. For filtering of genes that characterize developmental stages, a classification model created by a Random Forest algorithm was built. Using the importance of explanatory variables feature obtained from the created model, we identified 46 genes selected in the body and 169 genes selected in the head that were defined as the "most characteristic genes" during eel metamorphosis. Next, network analysis and subsequently gene clustering were conducted using the most characteristic genes and their correlated genes, and then 6 clusters in the body and 5 clusters in the head were constructed. Then, the characteristics of the clusters were revealed by Gene Ontology (GO) enrichment analysis. The expression patterns and GO terms of each stage were consistent with previous observations and experiments during the larval metamorphosis of the Japanese eel. CONCLUSION: Genome and transcriptome resources have been generated for metamorphosing Japanese eels. Genes that characterized metamorphosis of the Japanese eel were identified through statistical modeling by a Random Forest algorithm. The functions of these genes were consistent with previous observations and experiments during the metamorphosis of anguillid eels.

Short-term induced swimming activity enhanced innate immune parameters and antioxidant status of European eel (Anguilla anguilla).

The swimming activity, although an essential trait in the life cycle of fish, is still poorly understood in farmed fish. The current study aimed to investigate the impact of short-term induced swimming on the immune and antioxidant defence systems in European eel (Anguilla anguilla). Sixteen male yellow European eels (total length: 39.9 ± 0.7 cm; body weight: 108.8 ± 6.1 g) were individually placed in swimming flumes and divided into two groups: i) no swimming (n = 8); and ii) induced-swimming (n = 8) at 0.3 body lengths (BL)·s-1 for 7 h. Swimming resulted in a 2-fold lower cortisol concentration in plasma, whereas plasma glucose, lactate, and several immune-related parameters did not present variations between groups. Interestingly, swimming led to higher lysozyme, peroxidase, and protease activities in skin mucus, whereas bactericidal activity did not show differences among groups. Additionally, the gene expression of interleukin 1 beta showed an up-regulation in the skin of fish with induced swimming, while no differences were observed in the head-kidney or gills. Furthermore, modulation of the antioxidant status was observed in the liver and posterior skeletal muscle after induced swimming. Fish subjected to swimming showed lower lipid peroxidation and higher reduced glutathione levels, increasing the reduced/oxidized glutathione ratio. However, no variations in the antioxidant status were observed between groups in the anterior skeletal muscle. This study showed modulation of immune and oxidative stress markers in European eels upon short-term induced swimming compared to non-swimming fish.

Can displaced Anguilla marmorata return to their original habitat? Daily tracking study of their homing behavior.

Anguilla marmorata collected in the Odana River lower reaches were passive integrated transponder-tagged displaced and released into the upper river reaches (54 up-transported eels), and eels collected in the upper reaches were tagged and released downstream (52 down-transported eels). Their movements were detected once per day for 10 days using a portable radio-frequency identification (RFID) system. The homing rate of the down-transported eels was 38.9%, compared to 3.7% for the up-transported eels, suggesting that eels inhabiting upstream areas have relatively high fidelity to their habitats and downstream eels have less fidelity.

Preparation of valine-curcumin conjugate and its in vitro antibacterial and antitumor activity and in vivo biological effects on American eels (Anguilla rostrata).

Curcumin (Cur) exhibits diverse natural pharmacological activities, despite its limited water solubility (hydrophobicity) and low bioavailability. In this investigation, a valine-curcumin conjugate (Val-Cur) was synthesized through amino acid side chain modification, and its solubility increased to 1.78 mg/mL. In vitro experimental findings demonstrated that the antibacterial activity of Val-Cur against Escherichia coli, Staphylococcus aureus, Aeromonas hydrophila, and Vibrio parahaemolyticus was significantly superior to that of Cur. The inhibition rate of Val-Cur against HepG2 (human hepatocellular carcinoma) cells was higher than that of Cur at low concentrations (below 25 µmol/L), although the IC50 value of Val-Cur did not differ significantly from that of Cur. In vivo biological effects of Val-Cur were assessed by adding it into the feed (150 mg/kg) of American eels (Anguilla rostrata). Val-Cur significantly improved the growth performance (↑weight gain rate, ↑specific growth rate, and ↓feed conversion rate) and activities of intestinal digestive enzymes (amylase and lipase) and antioxidant enzymes (superoxide dismutase) in American eels. Additionally, Val-Cur significantly improved serum biochemical indices (↑high-density lipoprotein cholesterol, ↓low-density lipoprotein cholesterol, ↓aspartate and alanine aminotransferases). Furthermore, Val-Cur increased intestinal microbial diversity, reduced the abundance of potentially pathogenic bacteria (Spiroplasma, Clostridium, and Pseudomonas), and elevated the abundance of beneficial digestion-promoting bacteria (Romboutsia, Phyllobacterium, Romboutsia sedimentorum, and Clostridium butyricum) conducive to glucose metabolism (P < 0.05). To the best of our knowledge, this study is the first to explore water-soluble curcumin in aquaculture, and the findings will lay the groundwork for the potential application of water-soluble curcumin in the field of aquaculture.

Lipids as biomarkers to assess the nutritional and physiological status of two diadromous fish (Anguilla anguilla and Chelon auratus) at early life stages in a temperate macrotidal estuary.

Estuaries are considered as key habitats for the early life stages of fish. However, in the face of massive destruction of many estuarine intertidal areas, management and conservation measures are needed. Fish condition indicators may be used as a proxy of habitat quality and provide valuable information for management of coastal areas. In this study, the larvae of golden mullet (Chelon auratus) and European glass eels (Anguilla anguilla) were sampled in three sites of the Gironde Estuary. Different lipid classes and fatty acids were quantified: phospholipids (globally, phosphatidylethanolamine and phosphatidylcholine), triglycerides, omega-3 (particularly docosahexaenoic and eicosapentaenoic acids), omega-6 and C18:1. These biomarkers provide information on the nutritional status of the larvae as well as on prey availability and larvae diet between sites. One site significantly differed from the others as it seemed to offer abundant and better-quality prey. The very high levels of omega-3 contained in mullet larvae suggested that this site provided a high amount of diatoms. However, the mullet larvae that colonized this site also showed physiological stress that could be explained by exposure to pollutants through their prey. This work constitutes an essential baseline for developing biomarkers to assess the quality of habitats in a global change context.

IRF11 synergizes with STAT1 and STAT2 to promote type I IFN production.

Interferon regulatory factor 11 (IRF11), a fish specific member of IRF family, is a transcription factor known for its positive role in teleost antiviral defense by regulating IFN expression. Despite its recognized function, the precise mechanism of IRF11 in type I IFNs production remains largely unknown. In this study, we identified IRF11 in Japanese eel, Anguilla japonica, (AjIRF11) and determined its involvement in the later phase of fish IFN production. Our results demonstrate that IRF11-induced IFN production operates through ISRE binding. Mutations in each ISRE site within the promoter of AjIFN2 or AjIFN4 abolished IRF11-mediated activation of IFN promoters. In addition, the overexpression of AjIRF11 does not significantly impact the activation of AjIFN promoters induced by RLR-related signaling pathway proteins. Furthermore, IRF11-knockdown in ZFLs (zebrafish liver cells) has no effect on the RLRs-induced expression of zebrafish IFN-φ1 and IFN-φ3, indicating that IRF11 is not involved in the RLR-mediated IFN production. However, AjIRF11 can form transcription complexes with AjSTAT1 or AjSTAT2, or form homo- or heterodimers with AjIRF1 to stimulate the transcription of type I IFNs. Overall, it is shown in this study that IRF11 can act synergistically with STAT1 and/or STAT2 for the induction of IFN.

Effects of adding Bacillus subtilis natto NTU-18 in paste feed on growth, intestinal morphology, gastrointestinal microbiota diversity, immunity, and disease resistance of Anguilla japonica glass eels.

Japanese eel, Anguilla japonica, holds significant importance in Taiwanese aquaculture. With the intensification of eel farming, the impact of Edwardsiella tarda has become increasingly severe. Consequently, the abusive use of antibiotics has risen. Bacillus subtilis natto NTU-18, a strain of Bacillus with a high survival rate in feed processing, plays a crucial role in promoting intestinal health through competitive rejection, enhancing immune responses against bacterial pathogens, and improving intestinal health by modulating gastrointestinal microbiota to produce beneficial metabolites of mice and grass carp, Ctenopharyngodon idella. This study investigated the effects of different proportions (control, 0.25 %, 0.5 %, 1 %, and 2 %) of B. subtilis natto NTU-18 added to paste feed on the growth performance, intestinal morphology, and microbiota, expression of immune-related genes, and resistance to E. tarda in Japanese glass eel. The results indicated that the growth performance of all groups with B. subtilis natto NTU-18 added was significantly higher than that of the control group and did not impact the villi morphology. The expression of immune-related genes in the kidney, specifically HSP70 and SOD, was significantly higher from 0.5 % and above than the control; however, no significant differences were observed in CAT, POD, and HSP90. In the liver, significant differences were found in HSP70 and IgM above 0.25 % compared to the control group, with no significant differences in SOD, CAT, POD, and HSP90 among all groups. Additionally, intestinal microbiota analysis revealed that the 2 % additional group had significantly lower diversity than other groups, with Cetobacterium as the dominant species. The challenge test observed that the survival rates of the 0.5 % and 1 % groups were significantly higher. This research suggests that adding 0.5 % and 1 % of B. subtilis natto NTU-18 to the diet is beneficial for Japanese glass eel's immunity, growth performance, and disease resistance.

The expression profiles of cyp19a1, sf-1, esrs and gths in the brain-pituitary during gonadal sex differentiation in juvenile Japanese eels.

Eels are gonochoristic species whose gonadal differentiation initiates at the yellow eel stage and is influenced by environmental factors. We revealed some sex-related genes were sex dimorphically expressed in gonads during gonadal sex differentiation of Japanese eel (Anguilla japonica); however, the expression of sex-related genes in the brain-pituitary during gonadal sex differentiation in eels is still unclear. This study aimed to investigate the sex-related gene expressions in the brain-pituitary and tried to clarify their roles in the brain and gonads during gonadal sex differentiation. Based on our previous histological study, the control eels developed as males, and estradiol-17ß (E2) was used for feminization. Our results showed that during testicular differentiation, the brain cyp19a1 transcripts and aromatase proteins were increased significantly; moreover, the cyp19a1, sf-1, foxl2s, and esrs (except gperb) transcripts in the midbrain/pituitary also were increased significantly. Forebrain gnrh1 transcripts increased slightly during gonadal differentiation of both sexes, but the gnrhr1b and gnrhr2 transcripts in the midbrain/pituitary were stable during gonadal differentiation. The expression levels of gths and gh in the midbrain/pituitary were significantly increased during testicular differentiation and were much higher in males than in E2-feminized females. These results implied that endogenous estrogens might play essential roles in the brain/pituitary during testicular differentiation, sf-1, foxl2s, and esrs may have roles in cyp19a1 regulation in the midbrain/pituitary of Japanese eels. For the GnRH-GTH axis, gths, especially fshb, may be regulated by esrs and involved in regulating testicular differentiation and development in Japanese eels.

Recent insights into egg quality and larval vitality of the European eel Anguilla anguilla.

To date, the eel industry still depends on wild-caught juveniles that are grown to marketable size. There is an urgent need to close the eel life cycle in captivity to make aquaculture independent of the natural population. With this artificial reproduction protocol, yolk-sac larvae can be produced but egg quality may be impaired. Low survival rates and high deformity rates are frequently observed during the first week after hatching. Over the past four years, we have conducted studies with the aim to optimize the artificial reproduction protocol, thereby focussing on increasing egg and larval quality. Weekly carp or salmon pituitary extract (PE) treatment was successfully replaced with recombinant gonadotropins (rGTHs) to mature female eels and produce larvae. 17α,20ß-dihydroxy-4-pregnen-3-one (DHP) was replaced with upstream precursor progesterone (P) to induce the endogenous production of DHP by the female eel. DHP and P were found equally potent in inducing oocyte maturation and ovulation. The effects of antibiotics on larval survival and the occurrence of deformities were investigated. Antibiotic treatment increased survival and decreased the occurrence of deformities indicating bacterial infection as an important cause. A deformity determination key for young eel larvae has been developed that provides a framework of reference for larval deformities which will be instrumental with gaining insights on the reasons behind each larval deformity. These improvements of the artificial reproduction protocol and hatchery practices will contribute to the production of robust eel larvae that survive, grow and metamorphose into juveniles that will later be able to reproduce in captivity.