Effect of salinity on occurrence of notochord deformities in Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> larvae

One of the difficulties to rear Anguilla japonica larvae is the frequent occurrence of notochord deformities. We tested the effect of salinity on the occurrence of the deformities, because we have been using 50 % diluted seawater (50 % SW) for glass eel production, on the basis of the fact that intermediate salinity saves energy due to lower cost for osmoregulation and contributes higher survival and growth rates. We reared 6-day-old larvae in 50 and 100 % SW for 85 days and observed their morphology. The occurrence rate of deformed larvae, including kyphosis and scoliosis, was significantly higher in 50 % SW (35.8 %) than in 100 % SW (25.4 %), while survival rate was significantly higher in 50 % SW (69.8 %) than in 100 % SW (32.3 %) and growth in 50 % SW (mean body depth: 7.9 ± 5.3 mm) was better than in 100 % SW (6.8 ± 4.6 mm). We speculate that the most of severely deformed larvae could not survive in the tougher condition of 100 % SW, showing the lower occurrence of deformed larvae. Eventually, the yield of normal larvae after 85 days rearing was 1.9-fold higher in 50 % SW than in 100 % SW, implying that the advantage of 50 % SW for rearing eel larvae outweighs the risk of deformities.     

Anosmic Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> can no longer detect magnetic fields

ABSTRACT:   Magnetosensitivity in anosmic Japanese eel Anguilla japonica was examined by conditioning and electrocardiography. Five eels from a freshwater farm in Kagoshima, Japan, were made anosmic by injection of boiling petroleum jelly (130–140°C) into the nasal cavity, a technique widely used in studies on olfaction and olfaction-related behavior of eels. Another five eels were not injected with boiling petroleum jelly and served as controls. Conditioning consisted of exposure of eels to an imposed magnetic field of 192 473 nT. After complete conditioning of the eels, the imposed magnetic field was reduced from 192 473 nT to 12 663 nT during the tests. The control eels showed a significant conditioned response (i.e. slowing of the heart beat) to the magnetic field after one set of conditioning runs. The anosmic eels did not respond to the magnetic field, presumably due to damage of the olfactory organ from the jelly injection. The results suggest that Japanese eels have a magnetosensitive organ in or around the nares. At 1 month after the anosmia treatment, there was no visible trauma to the olfactory lamellae of anosmic eels.     

Predation on glass eels of Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> in the Tone River Estuary,Japan

Aquaculture of Japanese eel Anguilla japonica relies upon the natural recruitment of their glass eels (juveniles); however, predation that could influence glass eel recruitment remains unknown. In the present study, we aimed to elucidate the proportion of predation on A. japonica glass eels through stomach content analysis of predatory fishes collected in the estuary region of the Tone River system and its vicinity in Japan. Species of the predated glass eels were identified by DNA barcoding. A total of 270 predatory fishes of 15 taxa was collected over 2 years. The overall proportion of predation on glass eels, genetically identified as Japanese eel, was 0.7%, but this rose to 2.0% when data were limited to fishes caught during the peak months of glass eel recruitment. A glass eel was found in the stomach contents of a channel catfish Ictalurus punctatus, an invasive species in this river system, and a blackfin sea bass Lateolabrax latus. These fishes are therefore considered potential predators of A. japonica glass eels. However, as the proportion of predation was low, and the glass eels represented only small proportions of predator stomach contents, further investigation is needed for a better understanding of predation on A. japonica glass eels, and its effects on the early life stages of this endangered species.     

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.     

Occurrence of estuarine and sea eels <Emphasis Type="Italic">Anguilla japonica</Emphasis> and a migrating silver eel <Emphasis Type="Italic">Anguilla anguilla</Emphasis> in the Tokyo Bay area,Japan

Otolith microchemical analyses of the strontium (Sr) and calcium (Ca) concentrations in the eels Anguilla japonica and A. anguilla caught in Tokyo Bay were undertaken to reconstruct the eels’ migratory histories. A. japonica in the yellow stage (immature stage) were caught in a bay without any adjacent rivers or streams. A. anguilla was in the silver stage (early maturing stage), and the eel was confirmed to have just begun spawning migration to the open ocean from Tokyo Bay based on the otolith Sr:Ca ratios, which showed a typical catadromous life history with low Sr:Ca ratio values throughout the eel’s life after recruitment. The mean Sr:Ca ratios in A. japonica from the elver mark to the otolith edge indicated the eels belonged to several general categories of migratory histories, including sea eels (average Sr:Ca ratio ≥6.0 × 10⁻³) and estuarine eels (average Sr:Ca ratio 2.5 to 6.0 × 10⁻³) based on the criteria reported previously in A. japonica. All eels had a certain freshwater life period, although the period was highly variable among fish. These results indicate that A. japonica has a flexible pattern of migration, with the ability to adapt to various habitats and salinities.     

Development of an in vitro culture system for producing eel larvae from immature ovarian follicles in Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

To standardize conditions during the final maturation and ovulation of ovarian follicles from Japanese eel, we have developed a culture system for the production of fertilizable eggs from post-vitellogenic ovarian follicles in vitro. Post-vitellogenic ovarian follicles were incubated in culture medium supplemented with 17α,20β-dihydroxy-4-pregnen-3-one (DHP) with or without bovine serum albumin (BSA) to assess the effects of protein concentration. Eggs that ovulated during incubation were fertilized, and the remaining follicles were incubated in prostaglandin F_2α (PGF_2α) for a further 3 or 6 h before fertilization. Male eels were injected repeatedly with human chorionic gonadotropin. The quality of eggs obtained under the different culture conditions was evaluated after artificial fertilization in terms of hatching success. Hatching rates tended to decrease with increasing concentrations of BSA in the incubation medium in a dose-dependent manner. The addition of PGF_2α drastically increased the number of eggs that ovulated, but the rate of hatching was greatly decreased compared with eggs obtained earlier by DHP incubation alone. The larvae obtained from artificially fertilized eggs produced in vitro survived for 14 days without feeding. We conclude that in vitro culture systems thus have a great potential for the acquisition of good quality eggs under tightly controlled artificial conditions, culminating in the production of eel larvae.     

Rapid identification of eels <Emphasis Type="Italic">Anguilla japonica</Emphasis> and <Emphasis Type="Italic">Anguilla anguilla</Emphasis> by polymerase chain reaction with single nucleotide polymorphism-based specific probes

ABSTRACT:   Standard molecular techniques, such as sequencing and restriction fragment length polymorphism analysis after polymerase chain reaction (PCR) amplification are relatively complicated, and species identification can take a long time when using such techniques. We established a quick method, using PCR with species-specific TaqMan Minor Groove Binder (MGB) probes based on single nucleotide polymorphism (SNP) to distinguish the two eel species Anguilla japonica and Anguilla anguilla . This method can be used in processed products. Partial sequences of the mitochondrial 16S rRNA gene were compared between A. japonica and A. anguilla to design a primer pair common to both A. japonica and A. anguilla and probes specific to A. japonica and A. anguilla . Different fluorescence intensities were produced in two PCR microtubes each containing A. japonica - and A. anguilla -specific probes for one target sample. We observed the fluorescence intensity of PCR products in microtubes under ultraviolet transillumination, with similar results to those obtained by real-time PCR. Therefore, SNP-based PCR is a powerful tool for identifying materials of processed foods from either A. japonica or A. anguilla .     

iTRAQ proteomic analysis of salinity acclimation proteins in the gill of tropical marbled eel (<Emphasis Type="Italic">Anguilla marmorata</Emphasis>)

Osmoregulation plays an important role in the migration process of catadromous fish. The osmoregulatory mechanisms of tropical marbled eel (Anguilla marmorata), a typical catadromous fish, did not gain sufficient attention, especially at the molecular level. In order to enrich the protein database of A. marmorata, a proteomic analysis has been carried out by iTRAQ technique. Among 1937 identified proteins in gill of marbled eel, the expression of 1560 proteins (80 %) was quantified. Compared with the protein expression level in the gill of marbled eel in freshwater (salinity of 0 ‰), 336 proteins were up-regulated and 67 proteins were down-regulated in seawater (salinity of 25 ‰); 33 proteins were up-regulated and 32 proteins were down-regulated in brackish water (salinity of 10 ‰). These up-regulated proteins including Na⁺/K⁺-ATPase, V-type proton ATPase, sodium–potassium–chloride co-transporter and heat shock protein 90 were enriched in many KEGG-annotated pathways, which are related to different functions of the gill. The up-regulated oxidative phosphorylation and seleno-compound metabolism pathways involve the synthesis and consumption of ATP, which represents extra energy consumption. Another identified pathway is the ribosome pathway in which a large number of up-regulated proteins are involved. It is also more notable that tight junction and cardiac muscle contraction pathways may have correlation with ion transport in gill cells. This is the first report describing the proteome of A. marmorata for acclimating to the change of salinity. These results provide a functional database for migratory fish and point out some possible new interactions on osmoregulation in A. marmorata.     

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.     

Enhanced local recruitment of glass eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> in Oyodo River,Miyazaki and offshore environmental conditions in 2002

To examine the environmental factors controlling the inshore recruitment dynamics of Anguilla japonica in the Oyodo River, Miyazaki Prefecture, Kyushu, glass eel samplings were carried out using fyke nets during winter (November–March) of 1994–2014. The peak CPUEs (catch per unit effort) were observed between November and February, but differed from year to year. The yearly CPUE was extremely high in 2002, when the sea surface temperature (SST) in the offshore area of the Oyodo River was the lowest in winter of all the sampling years. The negative SST anomaly of less than ? 0.5 °C was sustained in the offshore area during the recruitment season in 2002, which was caused by two combined factors; low air temperature and the Kuroshio path. The oceanographic data showed that the dominant path of the Kuroshio was displaced eastward at 31°N in 2002, which was different from the average Kuroshio path. The eastward displacement of the Kuroshio induced a cyclonic mesoscale eddy in the offshore area of the Oyodo River, resulting in the entrainment of the cold seawater into coastal waters from deep water. The oceanographic condition in relation to the continuous low SST could be favorable for local recruitment of glass eels.     

A simple method to distinguish two commercially valuable eel species in Japan <Emphasis Type="Italic">Anguilla japonica</Emphasis> and <Emphasis Type="Italic">A. anguilla</Emphasis> using polymerase chain reaction strategy with a species-specific primer

In order to distinguish the two eel species Anguilla japonica and A. anguilla irrespective of unprocessed and processed samples, a quick, convenient method using polymerase chain reaction (PCR) with a species-specific primer was established. The comparison of partial sequences of the mitochondrial 16S ribosomal RNA gene between A. japonica and A. anguilla facilitated designing of a primer pair common to both A. japonica and A. anguilla and a primer specific to A. japonica. PCR was carried out with the three primers for total 110 specimens of A. japonica and A. anguilla. PCR products with the species-specific primer showed two bands for A. japonica and one band for A. anguilla in an agarose gel electrophoresis. This analytical procedure required only a short period and is more convenient than PCR-restriction fragment length polymorphism, one of the standard methods employed for fish species identification.     

Biomass fluctuation of two dominant lanternfish <Emphasis Type="Italic">Diaphus</Emphasis><Emphasis Type="Italic">garmani</Emphasis> and <Emphasis Type="Italic">D. chrysorhynchus</Emphasis> with environmental changes in the East China Sea

Acoustic surveys have been conducted for estimating the biomass of commercially important fish (e.g., anchovy, jack mackerel), lanternfish (Diaphus garmani and D. chrysorhynchus), and pearlside (Maurolicus japonicus) in summer in the East China Sea (ECS) since 1997. The biomass of lanternfish and pearlside was 2.26–19.16 times that of commercially important fish, and these species represented substantial biomass in the ECS. Though there were no correlations between biomass of pearlside and environmental indices, significant correlations between biomass of lanternfish and southern oscillation index (SOI) in March (positive correlation), arctic oscillation (AO) in March (negative) and October (positive), monsoon index (MOI) in February (positive), and Kuroshio flow mass in winter (positive) were observed. Weak AO and strong MOI would cool down the sea temperature and would lead to increased primary and secondary production in the ECS, thereby enhancing larval survival of lanternfish. The SOI would affect the Kuroshio meander in the ECS, and strong SOI and Kuroshio flow mass would transport larvae of lanternfish to the present survey area. This is the first report on the lanternfish standing stock and its fluctuation in the ECS.     

Spatial learning-induced <Emphasis Type="Italic">egr</Emphasis>-<Emphasis Type="Italic">1</Emphasis> expression in telencephalon of gold fish <Emphasis Type="Italic">Carassius auratus</Emphasis>

The immediate-early gene (egr-1) expression was used to examine the neuron’s response in telencephalon of goldfish during spatial learning in small space. Fishes were pre-exposed in the experimental apparatus and trained to pick food from the tray in a rectangular-shaped arena. The apparatus was divided into identical compartments comprising three gates to provide different spatial tasks. After the fish learned to pass through the gate one, two more gates were introduced one by one. Fish made more number of attempts and took longer time (P < 0.05) to pass through the first gate than the gate two or three. This active learning induces the expression of egr-1 in telencephalon as established by western blot analysis. Subsequently, the fish learn quickly to cross the similar type of second and third gate and make fewer errors with a corresponding decline in the level of egr-1 expression. As the fish learned to pass through all the three gates, third gate was replaced by modified gate three. Interestingly, the level of egr-1 expression increased again, when the fish exhibit a high exploratory behavior to cross the modified gate three. The present study shows that egr-1 expression is induced in the telencephalon of goldfish while intensively acquiring geometric spatial information to pass through the gates.     

Effects of environmental variables on recruitment of <Emphasis Type="Italic">Anguilla japonica</Emphasis> glass eels in the Yangtze Estuary,China

The recruitment pattern of Anguilla japonica glass eels into the Yangtze Estuary, China was studied during five successive migration seasons (2012–2016). The variation in daily catch was analyzed in relation to environmental factors that affect inshore migration. Two large migration waves of glass eels were observed in each recruitment season: one in the winter (January/February) and the other in the spring (March/April). A generalized additive model indicated that daily changes in glass eel catch were partially affected by local water temperature and tidal range. Water temperature seemed to have a threshold effect in winter, and there were optimal temperature ranges of 6–8 °C and approximately 10.5–12.0 °C associated with different recruitment waves to the Yangtze Estuary. The influence of tides was complex, with an overall positive correlation. The annual glass eel catch fluctuated greatly, but generally did not show a significant downward trend. To achieve sustainable use of glass eels, it is proposed that relevant management and conservation activities be reinforced.     

Effects of water temperature on early development of Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

ABSTRACT:   To determine an optimal temperature range for efficient production of healthy eel larvae Anguilla japonica , the effect of water temperature on hatching, survival, and deformity rates was examined. The early ontogeny of morphological features in this species by incubating eggs at five different temperatures (19, 22, 25, 28 and 31°C) was examined. Hatching occurred at 24, 28, 46 and 58 h after fertilization when incubated at 28, 25, 22 and 19°C, respectively; no eggs hatched at 31°C. The growth rate of prefeeding larvae increased as water temperature was elevated and acquisition of feeding ability was also accelerated at higher temperature. Significantly high hatching rates (76–86%, P  < 0.05) and survival rates (61–86%, P  < 0.05) were observed at relatively high temperatures (22–28°C). The deformities were 'open lower jaw', 'pericardial edema', and 'notochordal bending', in which the mouth was deformed with a downward projecting lower jaw, the pericardial cavity was swollen, and the notochord was bent or twisted to various degrees, respectively. Open lower jaw and pericardial edema were especially predominant at 19°C, with rates of 68 and 92%, respectively, compared with 31 and 10% at 25 and 28°C, respectively. The occurrence of notochordal bending was not affected by temperature. The optimal temperature for incubation and rearing A. japonica eggs and prefeeding larvae is approximately 25–28°C.