Mature Eels Research Articles

Effect of Salinity and Salmon Pituitary Extract on the Expression of Reproduction and/or Salinity-Related Genes in the Pituitary Cells of Japaneses Eel.

Artificial sexual maturation of eel (Anguilla japonica) involves rearing in seawater and injecting salmon pituitary extract (SPE). The salinity of seawater and components of SPE influence hormonal activities of the eel pituitary, leading to gonad development. This study investigated the direct effects of salinity change and SPE treatment on the eel pituitary gland using primary cell cultures. Pituitary cells were cultured into four experimental groups: control culture (control), SPE-treated culture (SPE), NaCl-treated culture (NaCl) and NaCl+SPE treated culture (NaCl+SPE). We investigated the expression of genes presumably related to reproduction and/or salinity, including luteinizing hormone (LHβ), follicle stimulating hormone (FSHβ), progesterone receptor-like (pgrl), prolactin (PRL), dopamine receptor D4 (drd4), neuropeptide B/W receptor 2 (NPBWR2) and relaxin family peptide receptor 3-2b (rxfp3-2b). Gene expression analysis revealed significant upregulation of LHβ in SPE and NaCl+SPE groups compared to control and NaCl (p<0.05). FSHβ expression did not show any significant changes. PRL showed a significant decrease in the NaCl group (p<0.05). Pgrl, NPBWR2, drd4, and rxfp3-2b displayed the highest expression in the control group, with downregulation observed in all treatment groups (NaCl, SPE, and NaCl+SPE) (p<0.05). This study demonstrated the direct effects of salinity changes and SPE treatment on the eel pituitary. Results from this study also suggest that salinity change is necessary but work together with SPE to induce reproductive process, and that LHβ, pgrl, PRL, drd4, NPBWR2, and rxfp3-2b genes are obviously associated with reproduction and salinity changes in eels.

Effects of dopamine and melatonin treatment on the expression of the genes associated with artificially induced sexual maturation in Japanese eel, Anguilla japonica.

Japanese eel (Anguilla japonica) is a commercially important fish species in Asia. Understanding factors like photoperiod, temperature, and lunar cycles is crucial for successful aquaculture and managing its reproduction. Melatonin and dopamine (DA) are essential for regulating reproduction in vertebrates, including fish. This study investigated the effects of melatonin and DA on the reproductive system of mature male Japanese eels to better understand reproductive regulation in fish. To clarify the effects of these hormones on sexual maturation in eels, a critical stage in the reproductive process, sexual maturation was induced by injecting human chorionic gonadotropin, which stimulates the production of sex hormones. To check the effect of melatonin and DA on sexual maturation, DA, melatonin, and DA + domperidone were intraperitoneally injected into fish from each group (six per treatment) at a dose of 1 mg/kg body weight. The fish were then examined using quantitative RT-PCR by comparing the messenger RNA level of reproduction-related genes (gonadotropin releasing hormone 1; gnrh1, gonadotropin releasing hormone 2; gnrh2, follicle stimulating hormone; fshβ, luteinizing hormone; lhβ and DA receptor 2b; d2b), involved in the gonadotropic axis in eels, to those that received a control injection. The results indicate significant differences in the expression levels of gnrh1, gnrh2 and d2b in the brain and d2b, fshβ, lhβ in the pituitary at different stages of sexual maturation. Melatonin appears to enhance the production of sex gonadotropins, whereas DA inhibits them. These findings suggest an interaction between melatonin and DA in regulating reproduction in Japanese eels.

Tissue-specific promoters regulate the transcription of cyp19a1 in the brain-pituitary-gonad axis of Japanese eel (Anguilla japonica)

Aromatase is a key enzyme that catalyzes the biosynthesis of estrogens. Previous study indicated that putative tissue-specific promoters of the one aromatase gene (cyp19a1) may drive the differential regulatory mechanisms of cyp19a1 expression in Anguilla japonica. In the present study, for elucidating the transcription characteristics and the function of putative tissue-specific promoters of cyp19a1 in the brain-pituitary-gonad (BPG) axis during vitellogenesis, we investigated the transcriptional regulation of cyp19a1 by 17β-estrogen (E2), testosterone (T), or human chorionic gonadotropin (HCG) in A. japonica. The expression of estrogen receptor (esra), androgen receptor (ara), or luteinizing hormone receptor (lhr) was up-regulated as cyp19a1 in response to E2, T, or HCG, respectively in the telencephalon, diencephalon, and pituitary. The expression of cyp19a1 was also upregulated in the ovary by HCG or T in a dose-dependent manner. Unlike in the brain and pituitary, the expression of esra and lhr, rather than ara, was upregulated by T in the ovary. Subsequently, four primary subtypes of 5′-untranslated terminal regions of cyp19a1 transcripts and the corresponding two 5′ flanking regions (promoter P.I and P.II) were identified. The P.II existed in all BPG axis tissues, whereas the P.I with strong transcriptional activity was brain- and pituitary-specific. Furthermore, the transcriptional activity of promoters, the core promoter region, and the three putative hormone receptor response elements were validated. The transcriptional activity did not change when the HEK291T cells co-transfected with P.II and ar vector were exposed to T. These results suggested that the expression of cyp19a1 was upregulated indirectly through esra and lhr rather than ara by T in the ovary, whereas the expression of cyp19a1 was upregulated directly through androgen receptor and the downstream androgen response element of tissue-specific P.I in the brain and pituitary. The results of the study reveal the regulatory mechanisms of estrogen biosynthesis and provide a reference for optimizing the technology of artificially induced maturation in eels.

North Equatorial Current and Kuroshio velocity variations affect body length and distribution of the Japanese eel Anguilla japonica in Taiwan and Japan

The larval stage of Japanese eel travels a substantial distance over a long duration through the North Equatorial Current (NEC) and the Kuroshio, and the spawning behavior of mature eels leads to monthly arrival waves in eastern Taiwan between November and February. The total length (TL) of the glass eel relates to its larval duration and age; therefore, the TL can indicate the larval duration. The monthly mean TLs of eels along eastern Taiwan from 2010 to 2021 were used to estimate the batch age, and the recruitment patterns and relative abundances were compared. The TLs of glass eels followed a normal distribution, and the estimated ages were highly correlated with their mean TLs. Early recruit TLs were significantly greater than those of late recruits. The mean tracer drift time was longer in early recruitment months (November–December) than in later dates (February–March). The recruitment lag between Taiwan and Japan was approximately 1–1.5 months, with relative more abundance in Taiwan for the early recruits and in Japan for the late recruits. Speculated cohorts followed the main streams of the NEC and Kuroshio, and the monthly velocity changes of these currents could affect the mean TLs as well as the distribution patterns of Japanese glass eels in Taiwan and Japan.

Do Metabolic Enzymes Explain Osmoregulation in Juvenile American Eels

During the spring, juvenile eels (Anguilla rostrata) (5-6 cm; 50-200 mg) gather in creeks along the Hudson River Tributary Estuary after migration from the Atlantic Ocean. This requires them to quickly adapt to environments with lower salt concentrations. We wondered how this is achieved. To study eels in a lab setting, we transferred them into Instant Ocean® sea salt (100% artificial sea water [ASW]) or dilutions thereof for 2 weeks; mortality was only observed in 100% ASW (37% expired in 3 hours). Interestingly, after 4 hours, 10 or 50% ASW eels showed little change in body mass, suggesting osmoregulation. While much progress has been made in understanding the mature eel response, few studies have reported enzyme activities in young eels. We hypothesized that dehydrogenases and acetylcholine esterase activities might provide a clue to explain osmoregulation and survival. Supernatants of whole juvenile eel extracts obtained from these experiments (IACUC-approved protocol) were run on 7.5% Tris-glycine PAGE gel (150V, 50 min) and the activities of non–denatured isozyme bands were analyzed with Bio-Rad ChemiDoc™ XRS+ Imager. Denatured supernatant extracts were also run on TGX 4-15% Tris-glycine PAGE gel to assess total protein banding. There was little difference in number of protein bands between eels exposed to 10% vs 50% ASW(2 and 3 weeks); however, band intensities were significantly higher for 50% ASW. Further studies revealed that two MDH (malate dehydrogenase) isozyme bands were present in extracts of eels exposed to a broad range of salinities: a relatively heavier band as mitochondrial (m) MDH and a lighter cytoplasmic (c) MDH. Activities of the mMDH bands were more robust than the cMDH bands at 50% and 10% ASW. Although the mMDH activities in eels exposed to either salinity were similar, the cMDH activities in eels exposed to 50% ASW were greater than in eels exposed to 10%. Additional experiments identified four acetylcholine esterase (AChE) isozyme bands in eel extracts; the activity of only one was significantly prominent in all exposures. Transferring the eels from 50% to 10% ASW reduced activity of the relatively heaviest isozyme band. Notably, we could not discern any fluctuations in lactate dehydrogenase (LDH) levels. In conclusion, while these preliminary findings do not reveal any concrete trends in metabolic regulation tied to survival, it does appear several key enzymes have a translational correlation tied to saline homeostasis. We acknowledge that the interpretation of these experimental results is complicated by a lack of genetic validation (i.e., sequences information and parental lines) - “nature” . In addition, the investigators were unable to track or define the environmental stresses faced during migration by individual juvenile eels before capture, which may manifest itself in unique stress responses - “nurture.”

Evaluating the impact of hydropower on downstream migrating anguillid eels: Catchment-wide and fine-scale approaches to identify cost-effective solutions

Hydropower is an increasingly popular source of renewable and ‘green’ (in terms of emissions) energy, but reduced longitudinal connectivity and diverting flow through turbines can have negative impacts on catadromous anguillid eel species that have declined globally. There is an urgent need for environmental managers to perform remediation actions, such as protecting flows for migratory fish and providing passage solutions at infrastructure, under increasing legislative pressure. To deliver this, a more comprehensive understanding of eel migration in catchments with hydropower is required. Here, we illustrate the importance of catchment-wide and fine-scale acoustic telemetry, coupled with the influence of eel maturation (i.e. sex steroid levels), to determine the impact of Wairua run-of-river Power Station (WPS) on downstream migrating shortfin eels (Anguilla australis; n = 25) in Wairua River, New Zealand. Migration speed through the unregulated reach upstream of WPS was positively correlated with flow, but not eel length or sex steroids. Three eels passed a diversion weir (DW) to follow the natural watercourse and eight entered the WPS canal. Eels predominantly entered (95.2%) and were last detected (85.7%) in WPS forebay during hours of darkness. Eleven (52%) of the 21 eels that entered WPS forebay were impinged or entrained, all when three or four turbines were in operation (power generation >3.04 MW). Ten (48%) passed WPS spillway during significantly higher spill than impinged or entrained eels, with four passing during no turbine operation, after experiencing high flows near the intake (multiple receivers in WPS forebay used to quantify fine-scale behaviour). On average, eels were impinged or entrained at WPS significantly quicker (6.40 ± 11.13 days) than eels that entered the spillway (25.17 ± 15.12 days), but eel length and sex steroids did not significantly influence fate. Of the eels that migrated through the entire 55 km study reach, passage time at DW and WPS equated to 0.01–0.02% and 47.62–92.17% of their migration, respectively. Mitigation for WPS (and similar power schemes) should focus on operational or physical changes at DW to minimise eels entering power station forebay(s). Turbine shutdowns, ensuring WPS spillway is available and the provision of a bypass channel in WPS forebay are also discussed as ways to conserve the species with the potential to save costs for water resource managers.

Evaluation of lethal and nonlethal assessment methods of muscle fat content in European eel (Anguilla anguilla)

Individual fat reserves are considered a key factor for the reproductive fitness of the endangered European eel (Anguilla anguilla). In contrast with most established standards, microwave measurements enable the determination of fat contents without sacrificing individual fish, offering a broad range of ecological applications. To test the reliability of nonlethal assessment methods of the muscle fat content in eels, the performance of microwave measurements was compared with the prevailing standard of measuring fat in a distinct subsample of muscle tissue by solvent extraction. Results indicate that either method is prone to error due to physiological and morphological changes during the sexual maturation of eels. Since microwave measurements were systematically affected by life stage and body length, it was possible to calibrate the method accordingly, putting it at least on par with the prevailing standard and further facilitating its use for scientific purposes.

Weaning European glass eels (Anguilla anguilla) with plant protein-based diets and its effects on intestinal maturation

Weaning glass eels with compound diets (36% proteins, 16% lipids) differing in their fishmeal (FM) level (50, 75 and 100% FM replaced by a blend of plant proteins, PP) was compared to a group fed cod roe. Weaning lasted for 20 days and then, eels were fed compound diets for 70 days, whereas the other group was only fed cod roe (90 days). Diets were tested with four replicates and evaluated in terms of growth, survival, glass eels metamorphosis into elvers, oxidative stress status and activity of digestive enzymes. Although glass eels are traditionally fed with fish roe and progressively weaned onto compound diets, results revealed that this strategy should not be prolonged for a long time, since feeding glass eels with cod roe for 90 days negatively affected their growth (2 times lower than fish fed compound diets), delayed their metamorphosis, as well as the maturation of their digestive function as the ratio of alkaline phosphatase and leucine-alanine peptidase indicated. Weaning glass eels onto compound diets differing in their FM levels did not affect their growth, metamorphic stage nor the activity of pancreatic enzymes (total alkaline proteases, trypsin, bile salt-activated lipase and α-amylase), although 75% FM replacement by PP sources delayed the level of intestinal maturation in eels. In comparison to glass eels fed the 100% FM diet, survival was negatively affected in groups fed diets with 50 and 75% FM replacement by PP ingredients, which indicated that further improvement is needed in diet formulation for this stage of development.

Long-term variation in numbers and biomass of silver eels being produced in two European river systems

Abstract The European eel (Anguilla anguilla) population has been in decline at least since the 1960s and reliable regional information, particularly on the spawner production and escapement (i.e. the silver eel life stage), is a requirement of the EU stock recovery regulation. Two comparable time series exist in Burrishoole (Ireland) and Imsa (Norway), with monitoring of total silver eel production since the early 1970s. Numbers of emigrating silver eels fell significantly (p &amp;lt; 0.0001) in the 1980s (breakpoints: Burrishoole 1982; Imsa 1988), in both catchments from &amp;gt;4000 eels per annum to ∼2000 eels per annum. The proportion of male eels dropped and the average size of female eels increased. Biomass of silver eels escaping has remained similar in Burrishoole (1.1/1.2 kg/ha), but not in Imsa (2.1/0.9 kg/ha) between the early period and the 2000s. Factors that govern the onset of eel maturation (silvering) and the annual production of silver eels are little understood. In this paper, the influence of time-lagged environmental variables on silver eel production is examined. Annual variation in the time series was partly (r2 Burrishoole = 0.43, Imsa = 0.46) explained by variation in water temperature and water level. Annual number of migrating eels in both catchments was positively related to summer temperature and summer water flow, negatively related to summer temperatures in the previous year, and in the Burrishoole, also negatively related to high water levels in September/October. The models did not transfer well between catchments, indicating likely catchment specific environmental factors impacting on eel production. The reduction in eel numbers observed in both catchments, accompanied by the change in sex ratio and mean weight of females that contribute to maintain biomass production, calls into question the advisability of basing a spawner escapement recovery target on biomass alone, while numbers and proportions of males decline.

Signal Transduction of Eel Luteinizing Hormone Receptor (eelLHR) and Follicle Stimulating Hormone Receptor (eelFSHR) by Recombinant Equine Chorionic Gonadotropin (rec-eCG) and Native eCG.

ABSTRACTPrevious studies showed that recombinant equine chorionic gonadotropin (rec-eCGβ/α) exhibits both follicle-stimulating hormone (FSH) and luteinizing hormone (LH)-like activities in rat LHR- and FSHR-expressing cells. In this study, we analyzed signal transduction by eelFSHR and eelLHR upon stimulation with rec-eCGβ/α and native eCG. The cyclic adenosine monophosphate (cAMP) stimulation in CHO-K1 cells expressing eelLHR was determined upon exposure to different doses (0–1,450 ng/mL) of rec-eCGβ/α and native eCG. The EC50 values of rec-eCGβ/α and native eCG were 172.4 and 786.6 ng/mL, respectively. The activity of rec-eCGβ/α was higher than that of native eCG. However, signal transduction in the CHO PathHunter Parental cells expressing eelFSHR was not enhanced by stimulation with both agonist rec-eCGβ/α and native eCG. We concluded that rec-eCGβ/α and native eCG were completely active in cells expressing eelLHR, similar to the activity in the mammalian cells expressing LHRs. However, rec-eCGβ/α and native eCG did not invoke any signaling response in the cells expressing eelFSHR. These results suggest that eCG has a potent activity in cells expressing eelLHR. Thus, we also suggest that rec-eCGβ/α can induce eel maturation by administering gonadotropic reagents (LH), such as salmon pituitary extract.

Eel Kisspeptins: Identification, Functional Activity, and Inhibition on both Pituitary LH and GnRH Receptor Expression.

The European eel (Anguilla anguilla) presents a blockade of sexual maturation at a prepubertal stage due to a deficient production of gonadotropins. We previously initiated, in the eel, the investigation of the kisspeptin system, one of the major gatekeepers of puberty in mammals, and we predicted the sequence of two Kiss genes. In the present study, we cloned and sequenced Kiss1 and Kiss2 cDNAs from the eel brain. The tissue distributions of Kiss1 and Kiss2 transcripts, as investigated by quantitative real-time PCR, showed that both genes are primarily expressed in the eel brain and pituitary. The two 10-residue long sequences characteristic of kisspeptin, eel Kp1(10) and Kp2(10), as well as two longer sequences, predicted as mature peptides, eel Kp1(15) and Kp2(12), were synthesized and functionally analyzed. Using rat Kiss1 receptor-transfected Chinese hamster ovary cells, we found that the four synthesized eel peptides were able to induce [Ca2+]i responses, indicating their ability to bind mammalian KissR-1 and to activate second messenger pathways. In primary culture of eel pituitary cells, all four peptides were able to specifically and dose-dependently inhibit lhβ expression, without any effect on fshβ, confirming our previous data with heterologous kisspeptins. Furthermore, in this eel in vitro system, all four peptides inhibited the expression of the type 2 GnRH receptor (gnrh-r2). Our data revealed a dual inhibitory effect of homologous kisspeptins on both pituitary lhβ and gnrh-r2 expression in the European eel.

Batch-like Arrival Waves of Glass Eels of Anguilla japonica in Offshore Waters of Taiwan.

Yu-San Han, Chau-Ron Wu, and Yoshiyuki Iizuka (2016) The larval stage of Anguilla japonica includes a long dispersal time over a long distance. In theory, the larvae should be distributed evenly throughout their transportation route when using both the NEC and Kuroshio, but the hypothesized new moon spawning of mature eels should lead to recruited glass eels exhibiting batch-like arrival waves, with a one-month-long cycle. However, environmental disturbances could mask the expected batch-like waves of glass eel recruitment. Thus, this phenomenon is best observed in glass eels collected from offshore waters, which are closer to the spawning site and less disturbed by these environmental factors. The offshore area of Yilan, Taiwan, is a suitable place to observe the arrival dynamics of the A. japonica glass eel. In this area, batch-like waves of glass eel arrival of A. japonica were observed, with peaks occurring between the last quarter and first quarter lunar periods, with a near one-month periodicity. No arrival peaks were found during the full moon period, suggesting that the glass eels exhibit light-avoidance behavior. Furthermore, all of the batches of arrivals were in the early pigmentation stage and similar in age (around 150-160 days), suggesting that they are likely a new arrival cohort. The tracer simulation showed that the mean tracer drift time, from the presumed spawning site to Yilan, was 155 ± 19.8 days. The observed batch-like arrival waves of glass eels in the offshore waters of Taiwan support the "New Moon Hypothesis," which suggests that there is synchronized spawning behavior of the eels during the new moon period.

Temperature modulates testis steroidogenesis in European eel

This study evaluates the effects of temperature on hCG-induced spermatogenesis in European eel (Anguilla anguilla), subjected to three thermal regimes: T10: 10°C (first 4weeks), 15°C (next 3weeks) and 20°C (last 6weeks); T15: 15°C (first 4weeks) and 20°C (last 9weeks); and T20: constant 20°C for the duration of the experiment. At 10°C, maturation stopped in the A spermatogonial stage (SPG1), and no further maturation was observed until the temperature was ≥15°C. With the aim of explaining these results, the influence of temperature on steroidogenic enzyme gene expression and steroid synthesis was tested. The initial synthesis of androgens (T and 11-KT) increased at SPG1, and was not influenced by temperature. Likewise, the gene expression of the steroidogenic enzymes linked to androgen synthesis (aacyp11a1, aacyp17-I and aa11βHSD) also increased at SPG1. In contrast, no correlation was seen between the increase in E2 and the aacyp19a1 gene expression peak in the testes, with E2 increasing as a consequence of the seawater acclimation carried out before hormonal treatment, and peaking the aacyp19a1 gene expression at B spermatogonial stage (SPG2). Aacyp21 gene expression was also higher at SPG2, and this stage was only reached when the rearing temperature was ≥15°C.In conclusion, androgen synthesis is not dependent on temperature, but further maturation requires higher temperatures in order to induce a change in the steroidogenic pathway towards estrogen and progestin synthesis. This study demonstrates that temperature plays a crucial role in European eel maturation, even perhaps controlling gonad development during the reproductive migration.

A question of origin: dioxin-like PCBs and their relevance in stock management of European eels

The stock of European Eel (Anguilla anguilla L.) has reached an all-time low in 2011. Spawner quality of mature eels in terms of health status and fitness is considered one of the key elements for successful migration and reproduction. Dioxin-like Polychlorinated Biphenyls (dl-PCBs) are known persistent organic pollutants potentially affecting the reproductive capability and health status of eels throughout their entire lifetime. In this study, muscle tissue samples of 192 European eels of all continental life stages from 6 different water bodies and 13 sampling sites were analyzed for contamination with lipophilic dl-PCBs to investigate the potential relevance of the respective habitat in light of eel stock management. Results of this study reveal habitat-dependent and life history stage-related accumulation of targeted PCBs. Sum concentrations of targeted PCBs differed significantly between life stages and inter-habitat variability in dl-PCB levels and -profiles was observed. Among all investigated life stages, migrant silver eels were found to be the most suitable life history stage to represent their particular water system due to habitat dwell-time and their terminal contamination status. With reference to a possible negative impact of dl-PCBs on health and the reproductive capability of eels, it was hypothesized that those growing up in less polluted habitats have a better chance to produce healthy offspring than those growing up in highly polluted habitats. We suggest that the contamination status of water systems is fundamental for the life cycle of eels and needs to be considered in stock management and restocking programs.

Repeated vertical movements of mature anguillid eels in a lake

Reproductive migration is a critical phase in the life history of anguillid eels. Nevertheless, fine-scale behaviours of migrating eels remain unknown, primarily due to the difficulty in attaching high-resolution recording devices to, and recovering them from, these small-sized teleosts. We attached a small accelerometer with time-scheduled release system to mature Anguilla celebesensis and A. marmorata in Lake Poso, Indonesia, during the pre-migration period. The eels repeated up-and-down movements in the water column (maximum depth, 77m), with slower, less active descents at shallower pitch angles, followed by faster, more active ascents at steeper pitch angles. The asymmetric diving pattern indicates negative buoyancy of the eels, which was confirmed by the measurements of body densities. The repeated diving is unlikely to represent foraging or thermoregulation because mature eels are thought to fast and water temperature changed little with depth. We suggest that the repeated diving is a result of the eels’ internal motivation for continuous swimming in preparation for oceanic migration, and is possibly energetically more efficient than if they keep swimming at a certain depth. The swimming energetics of eels in nature might be more complicated than previously thought because of the effect of vertical movements and non-neutral buoyancy.

Ultrasonography as a non-invasive tool for sex determination and maturation monitoring in silver eels

In the context of the severe decrease in temperate eel abundance, understanding and control of eel maturation has strong interest for scientific and commercial purposes. Possible use of ultrasonography for improvement of sex determination and maturation monitoring in silver eel was investigated. Gonads of 96 Anguilla anguilla silver eels were observed using portable equipment associated to a 6–15MHz probe, and sex determination was tried before artificial induction of maturation. To estimate gonad mass and monitor individual gonadosomatic index (GSI) in females, cross-sectional images were captured at different times of maturation and gonad length was measured at scanning. Two methods were tried for ovary mass estimation using ultrasonography: one based on a linear model and another on calculating ovary volume from a representation of gonad shape. Ultrasonography resulted in 100% success in sex determination. Ovary mass estimated by ultrasonography was strongly correlated to true ovary mass (R2=0.97). The use of a linear model for gonad mass and then GSI estimation seemed more appropriate than the use of a representation of gonad shape. Evolution of GSI estimates during maturation supports possible detection of early inter-individual differences in maturation using ultrasonography in female silver eels. This non-invasive tool can then obviously be exploited to improve sex determination in silver eels caught in the wild and to monitor maturation at the individual level. Ultrasonography thus has great potential for use in eel both for conservation and aquaculture. To our knowledge, this is the first report on the use of ultrasonography on eels or any anguillid species.

Effect of photoperiod on endocrine profiles and vitellogenin expression in European eels Anguilla anguilla during artificially induced ovarian development

The aim of this work was to determine the effects of dark and light conditions on the E2, testosterone and thyroid hormones levels and on the gene expression levels (vitellogenin 1, vitellogenin 2, and estradiol receptor one) in European eels (Anguilla anguilla) during ovarian development induced by increasing doses of carp pituitary extracts (CPEs). The subjects were divided into 2 groups: 14-hour light:10-hour dark (Light Group) and 24-hour darkness (Dark Group). All the eels received intramuscular injections with CPE at a dosage of 10 mg/kg body weight (BW) once a week for the first 3 weeks, 20 mg/kg BW fourth-sixth week, 30 mg/kg BW seventh-ninth week, and 40 mg/kg up to the end of the experiment (13th week). Vitellogenin and estradiol receptor expression levels did not show significant differences between the two housing conditions whereas in both groups vitellogenin mRNA increased starting from first CPE injection. Testosterone and 17-beta estradiol plasma levels were significantly greater in the Dark Group compared with the Light Group starting from the ninth and the 13th week, respectively. These results suggest that darkness could be a useful variable for standardizing gonadal maturation in eels kept in captivity.

Changes in the gene expression profiles of the brains of male European eels (Anguilla anguilla) during sexual maturation.

BackgroundThe vertebrate brain plays a critical role in the regulation of sexual maturation and reproduction by integrating environmental information with developmental and endocrine status. The European eel Anguilla anguilla is an important species in which to better understand the neuroendocrine factors that control reproduction because it is an endangered species, has a complex life cycle that includes two extreme long distance migrations with both freshwater and seawater stages and because it occupies a key position within the teleost phylogeny. At present, mature eels have never been caught in the wild and little is known about most aspects of reproduction in A. anguilla. The goal of this study was to identify genes that may be involved in sexual maturation in experimentally matured eels. For this, we used microarrays to compare the gene expression profiles of sexually mature to immature males.ResultsUsing a false discovery rate of 0.05, a total of 1,497 differentially expressed genes were identified. Of this set, 991 were expressed at higher levels in brains (forebrain and midbrain) of mature males while 506 were expressed at lower levels relative to brains of immature males. The set of up-regulated genes includes genes involved in neuroendocrine processes, cell-cell signaling, neurogenesis and development. Interestingly, while genes involved in immune system function were down-regulated in the brains of mature males, changes in the expression levels of several receptors and channels were observed suggesting that some rewiring is occurring in the brain at sexual maturity.ConclusionsThis study shows that the brains of eels undergo major changes at the molecular level at sexual maturity that may include re-organization at the cellular level. Here, we have defined a set of genes that help to understand the molecular mechanisms controlling reproduction in eels. Some of these genes have previously described functions while many others have roles that have yet to be characterized in a reproductive context. Since most of the genes examined here have orthologs in other vertebrates, the results of this study will contribute to the body of knowledge concerning reproduction in vertebrates as well as to an improved understanding of eel biology.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-799) contains supplementary material, which is available to authorized users.

Downstream migration dynamics of female and male silver eels (Anguilla anguilla L.) in the regulated German lowland Warnow River

AbstractIn the light of the European wide efforts to increase the spawning biomass of the European eel, a reliable measurement of the escapement of mature silver eel is necessary to prove the effectiveness of the conservation management measures. The seaward migration of mature eel is commonly viewed as a seasonal phenomenon with concentrated migration peaks occurring in spring and autumn. To verify the assumed seasonal silver eel migration events for regulated lowland rivers, a stow‐net system was installed in the Warnow River located in north‐eastern Germany. Between 2008 and 2011, the stow‐net system was operated from March to December each year. The eel harvest was documented on a weekly base including the documentation of weight and length, the silvering stage and the tissue sampling for the molecular identification of the eel species. During the 4 year monitoring period, a continuous downstream migration of female and male silver eels was observed. Additionally, single migration peaks were recorded in each year occurring between April and December. Moreover, female and male silver eels showed varying downstream migration dynamics. Based on a Chi‐squared Automatic Interaction Detection (CHAID) tree analysis, it was shown that during periods of a daily minimum air temperature over 10.4 °C, increased discharge levels and increased wind speeds, higher weekly migration rates of silver eels were likely. Furthermore, the results indicated that both sexes differed in their responses to migration triggering environmental factors. The presented results might be helpful to design more efficient eel conservation management strategies in regulated lowland rivers.

Quantitative bioassays for measuring biologically functional gonadotropins based on eel gonadotropic receptors

Significant declines in eel stocks have been noted in many parts of the world. Because eel aquaculture is dependent on wild-caught juveniles, there is a need to achieve artificial reproduction. Adult eel maturation is currently induced by repeated injections of purified gonadotropin (human chorionic gonadotropin [hCG]) or pituitary extract. Thus the determination of the biological efficacy and quantification of internal levels of gonadotropic hormones is important for optimizing artificial reproduction protocols. To quantify the plasma levels of biologically functional gonadotropic hormones, we developed a bioassay for luteinizing hormone (LH) and follicle-stimulating hormone (FSH) based on the stable expression of receptors in HEK293 cells of the Japanese eel Anguilla japonica LH (ajLHR) and the European eel Anguilla anguilla FSH (aaFSHR), respectively. Such cells also contain a firefly luciferase reporter gene driven by a cAMP-responsive element (CRE-Luc). We found that the obtained stable cells, with ajLHR, responded linearly to a more than 100,000-fold concentration range of hCG diluted in saline. The cells with aaFSHR showed a linear response to a 1000-fold concentration range of salmon pituitary extract mixed with saline. The biological functionality of the LH and FSH bioassays was validated using hCG, human FSH, and pituitary extracts from salmon, carp and eel. Since the toxins in eel plasma damaged the HEK293 cells, the protocol was adapted to selectively inactivate the toxins by heating at 37°C for 24h. This process successfully enabled the monitoring of hormone levels in blood plasma sampled from hCG-injected eels. In this paper, we describe the development of gonadotropin bioassays that will be useful for improving reproduction protocols in eel aquaculture.