Evidence for genetic differentiation in the european conger eel Conger conger based on mitochondrial DNA analysis

ABSTRACT:   The European conger eel Conger conger is an important marine benthic fish in the North-East Atlantic and represents a valuable fishery resource. However, little is known about its reproductive biology. In an attempt to gain a better understanding of the conger eel population structure, mitochondrial DNA (mtDNA) sequences were examined. A region with 432 bp of the control region of the mtDNA was sequenced from 40 individuals from six different locations around the central and eastern North Atlantic Ocean. Thirty variable positions defined 28 distinct haplotypes. The average sequence difference within samples (1.3–4.2%) was comparable to those between samples (1.4–3.6%). MtDNA sequence-based statistical tests showed significant geographic differentiation between some local population samples, suggesting that the conger eel does not comprise a single panmictic population. However, given our sample sizes, these preliminary results should be interpreted with caution and more individuals from more sites, including the Mediterranean Sea, should be analyzed in detail. The genetic variability detected in this study is an initial step to elucidate the genetic background of the conger eel population structure.     

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.     

Genetic structure of New Zealand eels Anguilla dieffenbachii and A. australis with allozyme markers

Abstract – Glass eels and adults of Anguilla australis and A. dieffenbachii were collected from rivers in the North and South Islands of New Zealand and tested for nine and ten allozyme loci, respectively. There was no heterogeneity among glass eel samples, but there was heterogeneity among adult samples within species. The results indicate that there may be single panmictic populations subject to sweepstake events or different selective pressures in the adult environments.     

Analysis of the genetic structure of the European eel Anguilla anguilla using the mtDNA D-loop region molecular marker

Sequence variation of the mtDNA D-loop region was analyzed in order to investigate the intraspecific evolution and the population genetic structure of the critically endangered European eel Anguilla anguilla. An additional attempt was made to collect information on the genetic characteristics of groups of eels representing naturally recruited eels to Lithuania and Latvia and introduced into Lithuanian lakes. A total of 148 eels were investigated, and 107 different haplotypes attributed to 39 haplogroups were determined during the study. A dataset comprising 229 sequences was created using the data from both this study and from earlier studies (n = 81). Analysis of this dataset revealed that haplotype diversity was 0.99567, the average number of nucleotide differences was 12.50544, there were 145 polymorphic sites and nucleotide diversity was 0.02426. No significant genetic differentiation was detected between different combinations of samples. However, the population genetic structure of this species could be characterized as a genetic mosaic formed due to the existence of reproductively isolated groups. The existence of a genetic mosaic in this species could be explained by the different evolutionary lineages found in the eel population.     

Developmental changes in pituitary–thyroid axis, and formation of gonads in leptocephali and glass eels of Anguilla spp.

SUMMARY: Pituitary, thyroid gland and gonads in leptocephali of Japanese eel Anguilla japonica (19.8–32.6 mm in total length), A. obscura (45.0 mm), and A. bicolor pacifica (49.5 mm) and those in glass eels of the Japanese eel were histologically and immunohistochemically examined in order to observe the developmental changes of these endocrine organs in the Anguillidae. The pituitary, consisting of adenohypophysis and neurohypophysis in Japanese eel leptocephali over 22.5 mm, did not contain thyroid stimulating hormone (TSH) immunoreactive cells. Such cells were, however, detectable in the more developed pituitaries of leptocephali of A. obscura and A. bicolor pacifica and in those of glass eels. Conversely, thyroxine (T₄)-immunoreactive thyroid follicles could be detected in all specimens, both leptocephalic and glass eel. Only in glass eels, gonads were found in the body cavity, and these gonads harbored one or two primordial germ cells (PGC) per cross-section. Our results indicate that thyroid hormones (TH) production started prior to TSH production, and that TSH and TH are both secreted during the metamorphosis from leptocephalus to glass eel. Therefore, it is plausible that the TSH–TH axis is involved in the metamorphosis from leptocephalus to glass eel, but not in the early growth from preleptocephalus to leptocephalus.     

Genetic divergence of kingfish from Japan, Australia and New Zealand inferred by microsatellite DNA and mitochondrial DNA control region markers

ABSTRACT: Genetic polymorphism in kingfish, collected from coastal waters of Japan, Australia and New Zealand, were examined using microsatellite (MS) DNA and mitochondrial DNA (mtDNA) control region markers. Sixteen to 25.7 alleles per locus were observed in three MS markers, while the average observed (and expected) heterozygosities were 0.782 (0.918), 0.750 (0.809) and 0.650 (0.888) for Australian, Japanese and New Zealand kingfish, respectively. Twelve mtDNA haplotypes were detected by the digestion of control region sequences with five endonucleases: Hae III, Hinf I Mbo I, Rsa I and Taq I. Significant genetic divergence was observed between the kingfish population from Japan and those from Australia–New Zealand. There was no significant differentiation among the Australian and New Zealand population samples.     

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

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

Presence of viruses in wild eels Anguilla anguilla L,from the Albufera Lake (Spain)

A virological analysis was conducted on wild eels from the Albufera Lake (Spain). A total of 179 individuals at different growth stages were collected in two different surveys (2004 and 2008). Presence of anguillid herpesvirus (AngHV‐1), aquabirnavirus and betanodavirus was confirmed by PCR procedures in both surveys, although the number of detections was clearly higher in 2008 (83% of the eels analysed resulted positive for virus presence). AngHV‐1 was the viral agent most frequently detected, followed by aquabirnaviruses. Betanodaviruses were detected by the first time in wild eels, and although the detections were only made by nested PCR, high percentage of positives were achieved. In addition, in 2008, seven aquabirnaviruses were isolated. Phylogenetic analysis performed using partial sequences of both genomic segments of aquabirnaviruses indicated that the seven isolates could be typed as WB (genogroup I) on the basis of segment A sequences, but when segment B was used six of them clustered with C1 strain (genogroup V) and one was typed as Ab (genogroup II). These results indicate natural reassortment between different strains of aquabirnaviruses in the eels. Although betanodaviruses were not isolated in cell culture, the analysis of the sequence of the nested PCR product indicated that they clustered with SJNNV genotype. The diversity of viral agents and the high level of viral detections suggest that viral infections may play a more prominent role in the decline of the European eel than initially thought.     

Larval Japanese eel (Anguilla japonica) as sub‐surface current bio‐tracers on the East Asia continental shelf

Larval Japanese eel (leptocephali) are passively transported from their spawning sites of the North Equatorial Current to the Kuroshio and its branch waters for 4–6 months before reaching the East Asian coasts. The larvae mainly stay within water depths between 50–150 m. The dispersal dynamics of larvae thus should reflect the sub‐surface oceanic currents on the East Asia continental shelf. An analysis of Japanese glass eel catch data in East Asian countries during 1985 to 2009, and for Taiwan from 1968 to 2008, indicates that the overall annual catch is generally correlated across countries of East Asia, and between north and west areas of Taiwan. The Kuroshio and its branch waters disperse glass eels throughout East Asian habitats, and the glass eel distribution matches the flow directionality of oceanic currents. Recruitment in western Taiwan occurs with a sequential southwestern to northwestern direction, suggesting that the Taiwan Strait Current penetrates the western coast of Taiwan in the sub‐surface layer in winter. The monthly averaged sub‐surface 50 m circulation pattern in the vicinity of Taiwan and modeled tracer experiments also support the northward winter sub‐surface current in Taiwan Strait. These results suggest that the larval Japanese eel could serve as a valuable bio‐tracer of sub‐surface currents, and the earlier recruitment dynamics of Japanese glass eels in Taiwan could be a good predictor for the subsequent catch in other East Asia areas.     

The sex-ratio reversal of the Japanese eel Anguilla japonica in the Kaoping River of Taiwan: The effect of cultured eels and its implication

The sex ratio of wild Japanese eels in the Kaoping River of southwestern Taiwan has been extremely skewed towards females in the recent years. However, the sex ratio skewed towards males after Typhoon Mindulle, July 2, 2004 then recovered to the previous female-dominated status in the following year. To determine why the sex ratio drastically changed, eels captured in the river were examined by both morphologic characteristics and otolith elemental signatures by solution-based inductively coupled plasma mass spectrometry (SB-ICPMS) and laser-ablation ICPMS (LA-ICPMS). Most of the eels collected in the river after the typhoon had a blue-gray colored back, with morphology and sex ratio similar to that of cultured eels, which differed from wild yellow eels which had a green colored back. The chemical signature in otoliths of eels with a blue-gray colored back was similar to that of cultured eels, with significantly lower Sr/Ca ratios but slightly higher Mn/Ca ratios than for wild eels. This confirmed that the reversal in eel sex ratio in the Kaoping River estuary resulted from cultured eels that had escaped from eel farms. Eel farmers estimated that about 30,000 eels escaped during the typhoon, sufficient to reverse the sex ratio of the eels in the river. Furthermore, silver eels caught in the estuary in the winter 2004 were also mostly males. The chemical signature in otoliths of these silver eels was similar to that of escaped cultured eels. Their morphology and mean GSIs, however, were comparable to wild silver eels. Thus, cultured eels that have escaped from eel farms can silver normally in the wild. Consequently, cultured eels may help to balance the sex ratio of the wild eel population and may contribute to the spawning stock of Japanese eel.     

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

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Antibody production in Japanese eels, Anguilla japonica Temminck & Schlegel

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

Late arrival of Anguilla japonica glass eels at the Sagami River estuary in two recent consecutive year classes: ecology and socio-economic impacts

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?C2010 and 2010?C2011 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?COctober. 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.     

日本鳗鲡产卵后生存及其繁育

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

Population structure of Japanese spiny lobster Panulirus japonicus inferred by nucleotide sequence analysis of mitochondrial COI gene

ABSTRACT:   The mitochondrial DNA sequence for the COI region was determined for 73 Panulirus japonicus individuals collected at three sample locations in Japan. Fifty-one haplotypes, including 69 polymorphic sites without insertion and deletion, were detected, of which three dominant haplotypes were shared among the three sample locations. The nucleotide and haplotype diversities at the three sample locations were in the ranges of 0.009–0.010 and 0.959–0.990, respectively. No significant population subdivision was detected among the sample locations based on the F _st value and net nucleotide substitution rate, and analysis of the molecular variance model ( amova ). There was no characteristic geographic distribution pattern for these haplotypes. These results indicate there is no genetic differentiation between P. japonicus populations in Japan, and support the hypothesis that benthic individuals of P. japonicus are sustained from a population of a common pool of long-period phyllosoma larvae through long-distance larval transport within the Kuroshio Subgyre.     

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

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

Genetic differentiation of the bronze featherback Notopterus notopterus between Mekong River and Tonle Sap Lake populations by mitochondrial DNA analysis

ABSTRACT:   Although the bronze featherback Notopterus notopterus is an important fish in South-East Asia, the population structure has not been investigated. In this study, genetic diversity and population structure of the bronze featherback were examined using nucleotide sequence analysis of the mitochondrial DNA control region for 332 fish collected from Tonle Sap Lake and the Mekong River in Cambodia. The average nucleotide diversity (π) of all samples was 0.033, and the Mekong River samples had higher nucleotide diversity (0.034) than Tonle Sap Lake samples (0.028). The nucleotide diversity between the lake and river samples varied from 0.029 to 0.037. The genetic differentiation between the river and lake populations was also supported by the pairwise F -statistic values and hierarchical analysis of molecular variance, indicating that the Tonle Sap Lake population is genetically isolated from the population in main stream of the Mekong River.     

基于线粒体COII和ND4基因序列的塔里木裂腹鱼群体遗传多样性分析

为探究塔里木裂腹鱼群体的遗传多样性和遗传分化现状,选用车尔臣河、克孜勒河和阿克苏河3个不同地理种群的塔里木裂腹鱼共计126尾样本,进行线粒体DNA COII和ND4基因序列的对比分析,探讨了2种标记对塔里木裂腹鱼遗传多样性分析结果的差异和关系。结果显示,塔里木裂腹鱼线粒体DNA COII和ND4基因序列的A+T含量均高于G+C含量,碱基组成具有偏倚性。基于线粒体DNA COII和ND4基因序列分析,126个样本中分别确定了6个和23个单倍型,其中线粒体DNA COII基因序列中3个群体存在共享单倍型现象,线粒体DNA ND4基因序列中未发现3个群体间存在共享单倍型。2种标记下群体的单倍型多样性和核苷酸多样性分别为0.738±0.019/0.904±0.014和0.02129±0.00298/0.04876±0.00149,呈现出高单倍型多态性和高核苷酸多态性的特征。2种标记的分子方差分析（AMOVA）均表明,在所有群体中遗传变异主要来源于群体间,且车尔臣河群体的塔里木裂腹鱼与其他2个群体的遗传分化均达到显著水平（P<0.05）。贝叶斯法（BI）构建的BI树与单倍型网络图结构一致,塔里木裂腹鱼形成了2个分支。COII和ND4基因序列的岐点分布图均呈现双峰型,表明塔里木裂腹鱼现在的分布是先前分化种群发生二次交流的结果。种群结构分析结果亦表明塔里木裂腹鱼已分化出2个明显的地理种群,故建议将车尔臣河群体作为塔里木裂腹鱼的亚种。     

An otolith microchemistry study of possible relationships between the origins of leptocephali of European eels in the Sargasso Sea and the continental destinations and relative migration success of glass eels

Martin J, Daverat F, Pécheyran C, Als TD, Feunteun E, Réveillac E. An otolith microchemistry study of possible relationships between the origins of leptocephali of European eels in the Sargasso Sea and the continental destinations and relative migration success of glass eels.
Ecology of Freshwater Fish 2010: 19: 627–637. © 2010 John Wiley & Sons A/S Abstract – Little is known about the extent to which Atlantic eels coming from different European rivers converge on the same spawning site. Our aim was to evaluate the spatial homogeneity of eel spawning area(s) with an otolith microchemistry approach. This work compared the elemental signatures of otolith’s core region of Anguilla anguilla leptocephali caught in the Sargasso Sea in 2007 with those of glass eels and elvers sampled in European estuaries during 2006, 2008 and 2009. Using laser ablation inductively coupled plasma mass spectrometry, the same annular ablation trajectory along the first feeding mark was applied on otoliths of glass eels, elvers and leptocephali. The concentrations of thirteen isotopes in the otoliths of glass eels/elvers did not vary among three annual cohorts collected in eleven European locations. However, otolith elemental fingerprints of leptocephali differed significantly from glass eels otolith’s signature. Although the mechanisms that regulate the differences in trace element signatures among leptocephali and glass eels/elvers are unknown, we propose that the sampled glass eels/elvers were born in a spawning site or region where favourable transport and/or feeding conditions occurred. Conversely, the leptocephali may have been sampled in a less favourable region in the Sargasso Sea, with a low probability of reaching continental growth areas.