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

The Japanese eel Anguilla japonica is an important aquaculture fish species in the East Asian countries of Japan, China, Korea, and Taiwan. All glass eel fry are captured from the wild and understanding the recruitment patterns of the glass eel is important. The larvae of A. japonica are passively transported to the East Asian Continental Shelf by the North Equatorial Current, the Kuroshio, the Kuroshio intrusion currents, and coastal currents. In each location, recruitment time is diverse: It is November in Taiwan and April in the Yalu River. How the glass eels reach recruitment areas remains poorly understood. Here, we combine information from larval ages based on otolith increments, simulated drifting paths on the East Asian Continental Shelf, and main fishing seasons in each location of East Asia. We identify five main recruitment blocks: (1) The main Kuroshio, (2) The Taiwan Strait Warm Current, (3) The Taiwan Warm Current, (4) The Yellow Sea Warm Current and (5) The branch of Yellow Sea Warm Current. The counted age of the glass eels is significantly underestimated for the later recruits, possibly due to the cessation of the otolith edge growth under low water temperatures. This study clarifies the eel&rsquo, s larval characteristics and transport mechanisms in the East Asia Continental Shelf, providing important information for its recruitment dynamics in the marine stage.

To determine the actual status of the recruiting glass eel stock of Anguilla japonica, we conducted a continuous monthly monitoring program for 2 years between November 2009 and October 2011 at the Sagami River estuary, Japan. A total of 114 and 372 A. japonica glass eels were observed in the 2009–2010 and 2010–2011 seasons, respectively. Recruitment patterns were the same in both years, starting in early winter (December in 2009 and November in 2010), increasing slightly until March, decreasing in April, but attaining the largest peak in June, after which no glass eels were observed during August–October. The Japanese eel has been known to spawn mainly in the summer, and glass eels recruit to their freshwater growth habitats during the winter to early spring. Our results clearly demonstrate an unexpected late arrival of glass eels in the early summer for two recent consecutive year classes. The summer recruitment found in our study indicates the unusual phenology of the Japanese eel, which may be a possible response to recent climate change.

Eggs of the Japanese eel Anguilla japonica collected in the western North Pacific were identified by onboard species-specific polymerase chain reaction (PCR) and DNA nucleotide sequencing after the cruise. Fish eggs of various species were collected by large plankton net tows at 12 stations along the southern part of the West Mariana Ridge on 19–25 May 2009. A total of 43 fish eggs were distinguished morphologically as possibly being of A. japonica. Thirty-one of those were analyzed by PCR, which included 15 eggs collected at 12°50–55′N, 141°15–20′E (in 5 tows) that showed positive results. The 16S ribosomal RNA (rRNA) gene sequences of eggs determined after the cruise indicated that 31 A. japonica eggs had been collected. The remaining eggs were of mesopelagic eel species (Serrivomeridae and Derichthyidae), or unidentified species. The morphological characteristics of the A. japonica eggs were consistent with those of artificially spawned eggs, except they had a slightly larger diameter. The egg diameter range did not overlap with those of mesopelagic eels of the Serrivomeridae, which often spawn in the same area as A. japonica. These results suggest that egg diameter and embryo shape can be used to morphologically identify naturally spawned A. japonica eggs.

The distribution of all larval stages of the Japanese eel, Anguilla japonica, were examined using historical catch records and original data in the western North Pacific (WNP) to evaluate existing information about the larval distribution and migration of this species. A total of 148 preleptocephali, 2547 leptocephali, 6 metamorphosing larvae, and 21 glass eels were collected during 37 cruises over a 52-year period (1956–2007). Sampling effort was spatio-temporally biased in latitude/longitude among seasons with sampling effort being concentrated near the western margin of the subtropical gyre near Taiwan in the winter season and extensive effort occurring near the spawning area to the east near the seamount chain of the West Mariana Ridge in summer during the spawning season. The distribution of preleptocephali (4.2–8.7 mm) was limited to a narrow area around 14°N, 142°E just west of the southern part of the seamount chain, while leptocephali (7.7–62.0 mm) were widely distributed at increasing size westward in the North Equatorial Current (NEC) to the region east of Taiwan. Metamorphosing larvae (52.7–61.2 mm) were collected only in the area 21–26°N, 121–129°E to the east of Taiwan, while glass eels (51.3–61.2 mm) occurred only within or west of the Kuroshio. These distributions suggest that leptocephali begin to metamorphose within or just east of the Kuroshio, then after completion of metamorphosis the glass eels detrain from the current and migrate inshore. The relationship between catch date and body size of leptocephali suggested that the spawning season is from April to August, but further sampling is needed to eliminate possible effects of sampling bias. This analysis is consistent with the existing hypothesis that Japanese eel larvae born near the West Mariana Ridge are transported westward in the NEC and then transfer to the Kuroshio to recruit to East Asia, although more sampling effort is needed for later stage larvae in the NEC bifurcation region to help understand the larval migration in relation to the possible impacts of ocean–atmosphere changes.

The Japanese eel (Anguilla japonica), European eel (Anguilla anguilla), and American eel (Anguilla rostrata) are migratory, catadromous, temperate zone fish sharing several common life cycle features. The population genetics of panmixia in these eel species has already been investigated. Our extensive population genetics analysis was based on 1400 Gb of whole-genome sequence (WGS) data from 84 eels. It demonstrated that a Japanese eel group from the Kuma River differed from other populations of the same species. Even after removing the potential adapted/selected single nucleotide polymorphism (SNP) data, and with very small differences (fixation index [Fst] = 0.01), we obtained results consistently indicating that panmixia does not occur in Japanese eels. The life cycle of the Japanese eel is well-established and the Kuma River is in the center of its habitat. Nevertheless, simple reproductive isolation is not the probable cause of non-panmixia in this species. We propose that the combination of spawning area subdivision, philopatry, and habitat preference/avoidance accounts for the non-panmixia in the Japanese eel population. We named this hypothesis the &ldquo, reproductive isolation like subset mapping&rdquo, (RISM) model. This finding may be indicative of the initial stages of sympatric speciation in these eels.

Minegishi, Y., Yoshinaga, T., Aoyama, J., and Tsukamoto, K. 2009. Species identification of Anguilla japonica by real-time PCR based on a sequence detection system: a practical application to eggs and larvae. – ICES Journal of Marine Science, 66: 1915–1918. To develop a practical method for identifying Japanese eel Anguilla japonica eggs and larvae to species by a sequence detection system using a real-time polymerase chain reaction (PCR), we examined (i) the sensitivity of the system using samples at various developmental stages, and (ii) influences of intra- and interspecific DNA sequence variations in the PCR target region. PCR amplifications with extracted DNA solution at 7.0 ng µl−1 or lower were efficient at distinguishing A. japonica from other anguillids. A single egg at the gastrula or later developmental stages could also be identified. Two sequence variations in the PCR target region were observed in 2 out of 35 A. japonica collected from three localities, and from four year classes at a single locality. These mutations, however, did not affect the result of species identification achieved by A. japonica-specific PCR primers and probe. The accuracy of this PCR-based method of species identification will help in field surveys of the species.

Extensive collections were made of the larvae of the temperate Japanese eel Anguilla japonica and the tropical giant mottled eel Anguilla marmorata in an overlapping area of the North Equatorial Current region of the western North Pacific Ocean. Collections of 189 A. marmorata and > 2500 A. japonica larvae during nine surveys from 1991 to 2007 showed that these two anguillid eels have similar spawning areas just west of the southern West Mariana Ridge. In July to August 2006 and August 2007, morphologically and genetically identified A. marmorata preleptocephali were mainly collected between 14.5-15 degrees N and 142-142.5 degrees E, where A. japonica preleptocephali were also caught in some of the same net tows. Fewer A. marmorata preleptocephali, however, were collected (n = 31) compared to those of A. japonica (n = c. 165), and fewer small larvae of A. marmorata were collected per tow than A. japonica (n = 1-10 and 1-294, respectively), suggesting relatively smaller spawning aggregations of A. marmorata. The distribution of preleptocephali and small larvae was wider in longitude in A. marmorata (131- 143 degrees E) than in A. japonica (137-143 degrees E), while the latitudinal range was almost the same (12-17 degrees N). Although spawning by these two species overlaps both spatially and temporally, the tropical eels of the North Pacific population of A. marmorata probably have a much longer spawning season with fewer spawners, at least in summer, and recruit to a much wider latitudinal range of growth habitats.

To compensate for the limited number of morphological characteristics of fish eggs and larvae, we established a convenient and robust method of species identification for eggs of the Japanese eel (Anguilla japonica) using a real-time polymerase chain reaction (PCR) that can be performed onboard research ships at sea. A total of about 1.2 kbp of the mitochondrial 16S ribosomal RNA gene sequences from all species of Anguilla and 3 other anguilliform species were compared to design specific primer pairs and a probe for A. japonica. This real-time PCR amplification was conducted for a total of 44 specimens including A. japonica, A. marmorata, A. bicolor pacifica, and 6 other anguilliform species. Immediate PCR amplification was only observed in A. japonica. We then tested this method under onboard conditions and obtained the same result as had been produced in the laboratory. These results suggest that real-time PCR can be a powerful tool for detecting Japanese eel eggs and newly hatched larvae immediately after onboard sampling during research cruises and will allow targeted sampling efforts to occur rapidly in response to any positive onboard identification of the eggs and larvae of this species.

The spawning area of the common Japanese conger, Conger myriaster, had remained unknown because spawning adults or its newly hatched larvae were never collected. Using genetic identification, we determined that C. myriaster spawns far offshore in the western North Pacific, just west of the spawning area of the Japanese eel, Anguilla japonica. In June 2008, six newly hatched C. myriaster larvae, 5.6–6.9 mm, were collected at the eastern edge of where many small unidentified Conger leptocephali (7–20 mm) were collected previously. The offshore spawning location of C. myriaster is analogous to that of the American conger eel, Conger oceanicus, and the American eel, Anguilla rostrata, in the Sargasso Sea, suggesting that convergent evolution of large-scale reproductive migration strategies in both anguillid and conger eels has occurred in the north Atlantic and Pacific subtropical gyres. The realization that two anguillids, A. rostrata and A. japonica, and two congers, C. oceanicus and C. myriaster, have evolved almost identical migration strategies in widely separated ocean basins suggests that natural selection for larval survival and recruitment success has resulted in long offshore spawning migrations in two phylogenetically distant taxa of anguilliform eels.

The natural reproductive ecology of freshwater eels remained a mystery even after some of their offshore spawning areas were discovered approximately 100 years ago. In this study, we investigate the spawning ecology of freshwater eels for the first time using collections of eggs, larvae and spawning-condition adults of two species in their shared spawning area in the Pacific. Ovaries of female Japanese eel and giant mottled eel adults were polycyclic, suggesting that freshwater eels can spawn more than once during a spawning season. The first collection of Japanese eel eggs near the West Mariana Ridge where adults and newly hatched larvae were also caught shows that spawning occurs during new moon periods throughout the spawning season. The depths where adults and newly hatched larvae were captured indicate that spawning occurs in shallower layers of 150–200 m and not at great depths. This type of spawning may reduce predation and facilitate reproductive success., Little is known about the reproductive ecology of freshwater eels. In this article, the authors describe the capture of two species of eels together with eggs and newly hatched larvae, and suggest that spawning takes place during the new moon at shallower depths than previously thought.

The Japanese eel Anguilla japonica spawns in the North Equatorial Current in the Philippine Sea and their larvae are transported to their freshwater growth habitats in East Asia. Here we report the occurrence and biological characteristics of glass eels of A. japonica that were collected on northern Luzon Island of the Philippines, the southern limit of the distribution range of this species. Anguillid glass eels recruiting to the Cagayan River estuary in January and February 2009 were collected and identified using their morphological and genetic characteristics. Among the 767 specimens, 52 glass eels collected in January were found to be A. japonica, while the remaining were tropical anguillid species. Age estimation revealed that the glass eels of A. japonica from northern Luzon Island were 147.2 ± 21.3 days old (mean ± sd) with a range between 111 and 185. The hatch dates of these A. japonica suggested that they were derived from at least four spawning events in the 2008 spawning season. Despite the increasing demand on the glass eels for aquaculture in the area where the five anguillid species simultaneously recruit, abundance of each species is yet unknown and investigations will be important for the protection of stock of the anguillids.