Characterization of microsatellite loci from the Japanese eel Anguilla japonica

The Japenese eel, Anguilla japonica, is generally assumed to be composed of a single population with wide distribution range, and some genetic studies using allozyme or mitochondrial DNA methods supported this population model. However, one genetic study suggested the existence of multiple populations in this species, and thus, more detailed studies on the population structure is needed. Here we characterized a total of 11 microsatellite markers of the Japanese eel. These will serve as powerful tools for detailed population study for the Japanese eel, though two of them showed the significant departure from the Hardy–Weinberg expectations.     

Recruitment mechanism of the eel, Anguilla japonica, to the Japanese coast

A total of 149 Anguilla japonica elvers collected at 10 locations along the Japanese coast were aged according to otolith daily increments. The interrelationships among age, birth date, body size, pigmentation stage, sampling location and the timing of recruitment were examined in order to determine the recruitment mechanism of elvers to coastal waters.
First catches of eels were earlier in the locations at lower latitudes. Age at recruitment was roughly constant, 218 ± 29 (mean ± s.d.) days old, disregarding the localities, but a weak positive correlation was obtained between age and sampling date or timing of recruitment. Body length at recruitment was also constant, 56.3 ± 2.3 mm s.l., and showed no significant correlation with either locality or recruitment timing. Estimated birth date ranged from April to November, the mean ± s.d. being 22 July 1982 ± 42 days, suggesting a peak season spawning in summer. Birth date was closely related with both the latitude of location and sampling time. Pigmentation developed more at lower latitude. Recruitment mechanism of the Japanese eel was summarized as follows: the earlier-born individual recruits earlier at lower latitude and at younger age, but at a constant body size.     

Internal illuminance and shelter shape affect shelter selection by the Japanese eel Anguilla japonica

The present study evaluated the effects of internal illuminance and shelter shape on shelter selection by Japanese eels to enable the preservation or construction of suitable shelters for the Japanese eel. Japanese eels were able to distinguish a 1.25-fold difference in illumination inside the shelters, and preferred darker shelters. When the internal illumination of two shelters with the same shape was less than one-tenth of the ambient illumination (about 400 lx), shelter selection by Japanese eels was not affected by internal illuminance, even when there was a 10-fold difference in internal illumination between shelters. The width of the shelter was not important, but Japanese eels preferred a deep shelter with a low ceiling and walls that spread to a ‘dead end’. This has important implications on the creation of suitable shelters for Japanese eels.     

Wavelength-specific thresholds of artificially reared Japanese eel Anguilla japonica larvae determined from negative-phototactic behaviours

We report wavelength-specific thresholds of leptocephali of Japanese eels Anguilla japonica determined from their negative-phototactic behaviour. Leptocephali are most sensitive to wavelengths 400–500 nm and at very short wavelengths. Their visual sensitivity decreases more sharply at wavelengths >500 nm than it does at wavelengths <400 nm. The spectral sensitivity of leptocephali adapts to the optical conditions of their habitat. The mean visual sensitivity threshold of leptocephali is 7.22 × 10⁻⁴ μmol m⁻² s⁻¹ between 400 and 500 nm. Based on visual sensitivity thresholds of 475 nm, the most transparent wavelength in waters where these leptocephali occur, the daytime depth of occurrence of these larvae may exceed 250 m. LEDs emitting light of wavelength 625 nm in culture environments would minimise disturbance to leptocephali during facility maintenance.     

Pseudodactylogyrus kamegaii sp. n. (Monogenea: Pseudodactylogyridae) from wild Japanese eel,Anguilla japonica

The monogenean Pseudodactylogyrus kamegaii sp. n. is described, based on specimens collected from the gills of wild Japanese eel Anguilla japonica caught in Chiba Prefecture, Japan. This species is the most similar to P. anguillae (Yin and Sproston, 1948), but different in the shape and measurements of the male copulatory organ, vagina and marginal hook. This new species was collected from the eel in brackish waters, while P. anguillae and P. bini, the other known pseudodactylogyrids of Japanese eel, have been recorded only in fresh waters.     

Temperature effects on the incorporation of strontium in otolith of Japanese eel Anguilla japonica

The effects of temperature on somatic and otolith growth and the incorporation of strontium in otolith of the Japanese eel, were studied in laboratory-reared and field-caught eels. The somatic and otolith growth rates of the eel increased significantly with temperature and were estimated as approximately 0·096 mm t.l, (P<0·01) and 0·36 μm in otolith diameter per degree-day (0·01相似文献   

Evaluation of downward movements of Japanese eel Anguilla japonica inhabiting brackish water areas

This study evaluated the size and age distributions and otolith microchemistry of the Japanese eel Anguilla japonica in freshwater and brackish water areas in the Aki and Tsuchikawa rivers for 1 year, and in brackish water areas in the Asahi River for 3 years to understand the movements of Japanese eels between continental habitats of different salinity after recruitment (n = 759). For all three rivers, the total length (L_T) and age distributions were consistent; yellow eels captured in the upper brackish water (Aki River: 353.5 ± 77.4 mm and 3.0 ± 0.8 years; Tsuchikawa River: 287.7 ± 87.3 mm and 3.7 ± 1.3 years; Asahi River: 418.2 ± 112.1 mm and 4.2 ± 1.7 years) were smaller and younger than not only those in the fresh water of the two rivers but also those in the lowest brackish water sampling areas (Aki River: 436.0 ± 71.6 mm and 3.8 ± 1.1 years; Tsuchikawa River: 370.9 ± 121.7 mm and 4.9 ± 2.3 years; Asahi River: 558.5 ± 85.9 mm and 5.7 ± 1.7 years). In the Asahi River, these tendencies were found throughout the 3 years. Otolith analysis indicated that the majority of the eels captured in the lowest brackish water areas had moved down from upstream. These results suggest that Japanese eels inhabiting saline water generally move from the upper estuary as they grow. The upper estuary can be an important area for the management of this species because these eels spend their early continental growth life there.     

Anguilla japonica is already magnetosensitive at the glass eel phase

Magnetosensitivity of the Japanese eel Anguilla japonica at the glass eel phase (newly metamorphosed juveniles) was examined by conditioning and electrocardiography. The glass eels were conditioned to an imposed magnetic field of 192 473 nT parallel to the fish body placed along the earth's west‐east axis. After 10 to 40 conditioning runs, all the glass eels exhibited a significant conditioned response ( i.e . slowing of the heart beat) to a 192 473 nT magnetic field and even to a 12 663 nT magnetic field that combined with the geomagnetic field (32 524 nT) at the laboratory and produced a resultant magnetic field of 21° easterly. These results indicate that glass eels have high magnetosensitivity and probably acquire geomagnetic information early in life. It is hypothesized that silver‐phase adult eels find their way back to the oceanic spawning ground by reversing the geomagnetic direction that had been detected and 'memorized' during the glass eel phase when migrating from the open ocean towards the continental shelf and coastal waters.     

Unravelling the life-history patterns and habitat preferences of the Japanese eel (Anguilla japonica) in the Pearl River,China

Eels have fascinated biologists for centuries due to their amazing long-distance migrations between freshwater habitats and very distant ocean spawning areas. The migratory life histories of the Japanese eel, Anguilla japonica, in the waters of south China are not very clear despite its ecological importance, and the need for fishery regulation and management. In this study, strontium (Sr) and calcium (Ca) microchemical profiles of the otoliths of silver eels were measured by X-ray electron probe microanalysis based on data collected from different habitats (including freshwater and brackish habitats), in the large subtropical Pearl River. The corresponding habitat preference characteristics were further analysed using redundancy analysis (RDA). A total of 195 Japanese eels were collected over 6 years. The collected individuals ranged from 180 to 771 mm in total length and from 8 to 612 g in body weight. Two-dimensional pictures of the Sr:Ca concentrations in otoliths revealed that the A. japonica in the Pearl River are almost entirely river eels, spending the majority of their lives in fresh water without exposure to salt water, while the catadromous migration time has delayed about 1 month in the Pearl River estuary in the past 20 years. RDA analysis further indicated that juveniles and adults preferred water with high salinity and high tide levels. Youth preferred habitats with high river fractals. Our findings contribute to a growing body of evidence showing that the eels are extremely scarce currently and conservation measures against them are imminent, including the protection of brackish and freshwater areas where they live in south China.     

鳗鲡病原性维氏气单胞菌的分离与鉴定

从养殖患病的日本鳗鲡(Anguilla japonica)肾脏分离得到1株优势菌株,形态特征和生理生化测试结果显示,分离菌在4%羊血平板培养基上呈α型溶血,革兰氏阴性,有运动性,氧化酶阳性,在pH6.0和1%NaCl中生长等。结合Biolog自动微生物鉴定系统和16S rRNA、gyrB基因序列分析结果,确定该分离菌为维氏气单胞菌(Aeromonas veronii)。人工腹腔注射感染鳗鲡的试验结果表明,分离菌对鳗鲡的半数致死量LD50为2.15×107CFU/mL。药敏试验结果显示,在13种抗菌类药物中,分离菌对左氧氟沙星、氟哌酸、阿奇霉素和萘啶酸等8种药物敏感,而对利福平和新生霉素等5种药物敏感度低。     

Determination of quantitative parameters of Escherichia coli phagocytosis by mouse peritoneal macrophages

Quantitative parameters of phagocytosis of fluorescein-labeled Escherichia coli cells by mouse peritoneal macrophages were studied using a fluorimetric method. E. coli cells were conjugated with fluoresceinisothiocyanate (FITC) and then incubated with macrophages. At the end of incubation, phagocytosis was stopped by the addition of a lysing solution (0.5% Triton X-100 in 0.01 M phosphate buffer in 0.15 M saline, pH 7.4). Trypan blue at a concentration of 0.04% was used as a quenching agent to differentiate between attached and ingested E. coli cells. It was shown that phagocytosis of E. coli cells depended on temperature and opsonization of bacteria. The number of E. coli cells ingested by macrophages increased rapidly for the initial 60 min of incubation at 37°C. To achieve optimal uptake of E. coli cells, their opsonization with 5% native serum was needed. The uptake of nonopsonized bacteria by macrophages was significantly lower than that of the opsonized ones (p < 0.05). Sodium azide was shown to produce a dose-dependent suppression of phagocytosis of E. coli cells by mouse peritoneal macrophages.     

Relative abundance, sex ratio and population structure of the Japanese eel Anguilla japonica in the Tanshui River system of northern Taiwan

From the Gong-Shy-Tyan (GST) Stream and the Tanshui River, Taiwan, eels ranged from 5.53 cm t.l . (elvers) to 72.5 cm t.l . Anguilla japonica accounted for 93-99%, A. marmorata 1-7% and A. bicolor pacifica less than 1% of all eels caught. Mean eel lengths increased from 10 cm t.l . at the downstream sites to 50 cm t.l . at the upstream sites. Females made up 92.8% of the sex-determined eels. Population density, averaged approximately 0.14 eel m⁻² (2.42 g m⁻²) and 0.25 eel m⁻² (0.92 g m⁻²) in downstream sites of the GST and Tanshui River, respectively, and decreased substantially with upstream distance. Eels were rarely found in the heavily polluted and dam constructed areas in the midstream site of the Tanshui River.     

Effects of changes in environmental calcium on prolactin secretion in Japanese eel,Anguilla japonica

Effects of changes in environmental Ca²⁺ on the secretion of prolactin, a possible hypercalcemic hormone, were examined both in vivo and in vitro in the Japanese ecl, Anguilla japonica. Transfer of seawater- or freshwater-adapted fish to fresh water, fresh water containing 10 mmol Ca²⁺ · 1^-1 sea water, Ca²⁺-free sea water, or deionized water was accompanied by significant changes in plasma Ca²⁺ levels after 7 days, except for the fish transferred from fresh water to fresh water and from sea water to sea water. Changes in external Ca²⁺ concentrations did not affect plasma prolactin levels, although plasma prolactin levels as well as pituitary prolactin contents were significantly greater in fish in a hypotonic environment than those in a hypertonic environment, regardless of the external Ca²⁺ concentration. Hypercalcemia, induced by removal of the corpuscles of Stannius, did not alter plasma prolactin levles. Incubation of the pituitary in the medium with different Ca²⁺ concentrations (up to 2.9 mmol·l^-1) did not affect the basal release of prolactin, except at an extremely low Ca²⁺ concentration (less than 0.1 mmol·l^-1) where prolactin release was inhibited. Addition of Ca²⁺ ionophore (A23187) to the medium led to a marked and significant increase in prolactin release, indicating that an increase in intracellular Ca²⁺ stimulates prolactin release. However, the effect was not specific to prolactin cells; a similar increase was seen in growth hormone release. These results indicate that changes in environmental Ca²⁺ concentration may not be the primary factor influencing prolactin secretion in the eel; changes in environmental osmolality or Na⁺ levels seem to be more critical for the regulation of prolactin secretion.Abbreviations CSX stanniectomy - DMSO dimethylsulphoxide - DW deionized water - FW fresh water - GH growth hormone - PRL prolactin - SW sea water     

Population structure of the Japanese eel, Anguilla japonica

Mitochondrial DNA (mtDNA) sequences that include (a) a part of the cytochrome b gene, (b) two tRNA genes, and (c) a part of the noncoding D-loop region of 31 Anguilla japonica (Japanese eel) and 1 A. marmorata collected from Taiwan, Japan, and mainland China were determined to evaluate the population structure of Japanese eel. Among 30 genotypes identified from the 31 Japanese eel mtDNAs sequenced, there are 58 variable sites, predominantly clustered at the D-loop region. The phylogenetic tree constructed by the unweighted pair-group method with arithmetic mean shows neither significant genealogical branches nor geographic clusters. Furthermore, the sequence-statistics test reveals little, if any, significant genetic differentiation. These results indicate that the 31 Japanese eels might come from a single population. Analysis of sequence variation in mtDNA by using the relationship between the number of segregating sites and the average number of nucleotide differences under the neutral mutation hypothesis reveals that neutral mutation acts as a major factor influencing the evolutionary divergence of the Japanese eel mitochondrial genome sequenced, especially in the noncoding region.      

Atomic force microscopy of peritoneal macrophages after particle phagocytosis

The atomic force microscope was used to image peritoneal macrophages after phagocytosis of latex beads with 0.45 m in diameter and of zymosan particles. The rigidity of the phagocytosed material allowed to image the live membrane at forces below 2 nn. Repeated scanning of the membrane unavoidably caused the protrusion of the beads and increased their virtual height. The influence of fixation by glutaraldehyde on the image and the corresponding force vs. distance curves were analyzed and compared. Short treatment with Triton X-100 enabled us to identify intracellular components, such as embedded latex beads, cell nucleus and cytoskeletal strands. The data demonstrate that it is possible to image living cells if they are bolstered by stiff material.The authors wish to thank Dr. H. Oberleithner for his generous support, helpful discussions and the suggestion of Triton treatment. The authors gratefully acknowledge Dr. I. Jahns for establishing the AFM technique at the Institute of Physiology. The work was supported by the Deutsche Forschungsgemeinschaft Projekt No. La 315/4-1.     

Age and growth of eels Anguilla japonica in a Chinese river

Age and growth of Anguilla japonica sampled from the Qiulu River in southern China were studied. Mean body lengths and weights of the eels at each age were back-calculated from their otoliths. The growth parameters for the von Bertalanffy function gave L _∞= 98–2cm, K =0·07 and t ₀=− 1·94. Scales were observed to form firstly at the middle part of the lateral line, at a minimum body length of 160 mm and were inappropriate for ageing.     

Polymorphisms of GA/GT Microsatellite Loci from Anguilla japonica

Six novel microsatellite loci, containing (GA)_15\N17 or (GT)_10\N19 perfect tandem repeats, were isolated and characterized for the catadromous eel Anguilla japonica. The allelic size of the 6 loci ranged from 79 to 226 bp in length. All loci were polymorphic with a mean number of 14.7 alleles per locus and a mean heterozygosity of 0.67, suggesting higher polymorphism than that of freshwater and anadromous fishes, but lower than that of marine fishes. Genotype diversity of the 6 loci ranged from 0.22 to 0.61 with a mean value of approximately 0.5. Cross-species amplification showed that 5 of the 6 microsatellite primers proved to be useful in addressing questions of population genetics for all Anguilla species. Received October 11, 2000; accepted December 29, 2000.     

Purification and characterization of a novel imidazole dipeptide synthase from the muscle of the Japanese eel Anguilla japonica

We have purified a novel enzyme from eel white muscle which catalyzes the syntheses of imidazole dipeptides, such as carnosine (β-alanyl-l-histidine), anserine (β-alanyl-π-methyl-l-histidine), and balenine (ophidine; β-alanyl-τ-methyl-l-histidine), directly from their precursors. The enzyme was purified 1130-fold from eel muscle by a series of column chromatographies. Although eel muscle contains a large amount of carnosine and only trace amounts of anserine and balenine, the anserine synthesizing activity was by far the highest. From gel permeation chromatography, the molecular mass of the enzyme was calculated to be 275 kDa. SDS-PAGE of the purified enzyme represented a band around 43 kDa, suggesting that the native enzyme is a hexamer or heptamer. The optimal pH and temperature were around 9.5 and 60 °C, respectively. K_m values for β-alanine and π-methyl-l-histidine were 44 and 89 mM, respectively. The enzyme was greatly activated by Zn²⁺ and inhibited by EDTA. The N-terminal amino acid sequence of 25 residues of the purified enzyme showed 52% amino acid identity to 38–62 residues of zebrafish haptoglobin precursor. The purified enzyme also exhibited hydrolytic activity against these imidazole dipeptides.