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.     

日本鳗鲡人工繁殖技术研究最新进展

日本鳗鲡（Anguilla japonica）人工繁殖是至今尚未解决的世界性难题。2001～2003年日本水产综合研究中心在实施政府资助研究专项“生态保护型增养殖体系建设一种苗生产、放流技术开发”期间,对已有40余年研究历史的日本鳗鲡人工繁殖涉及的亲鱼选育、催熟、催产、优质卵判别、仔鱼培育等关键技术进行了比较系统的研究。经过270d培育,于2003年首次成功获得日本鳗鲡白仔鳗苗（体长52．6mm）。本文在主要介绍这一研究成果的同时,概要介绍我国在这一领域的最新研究进展,希望能为我国的日本鳗鲡人工繁殖研究提供参考。     

Genetic variability of the Japanese eel Anguilla japonica (Temminck & Schlegel) related to latitude

Abstract— Isozyme genotypes of 400 glass eels recruiting to 4 localities along the east Asian coast, stretching from Taiwan to the Yalu River of northeastern China, were studied using starch gel electrophoresis. Geographic cline was found to exist in two loci: NADP-isocitrate dehygenase-1 and 6-phosphogluconate dehydrogenase. Frequencies of the most common allele of these two loci increased from south to north. In the latitudinal range of 25°N to 40°N, the magnitude of difference of IDH100 and PGD100 was 13% and 9% respectively. However, deviations from Hardy-Weinberg equilibrium were found to be insignificant in both loci at three of the four localities. The cline was, therefore, unlikely to have resulted from selection. Migration time-lag from different parts of the continent to the spawning ground in the western Pacific was suggested to be a possible reason for the formation of the cline.     

Hen egg yolk and skinned krill as possible foods for rearing leptocephalus larvae of Anguilla japonica Temminck & Schlegel

Usual diets for rearing leptocephalus larvae of Japanese eel Anguilla japonica include eggs of the endangered spiny dogfish Squalus acanthias (SE). We investigated the effects of alternative food materials, hen egg yolk (HEY) and exoskeleton‐free (skinned) Antarctic krill (SAK), on the growth and survival of eel larvae. We found that feed comprising whole krill including exoskeleton (WAK) containing higher levels of fluoride (37.89 mg kg^?1) was acutely toxic to eel larvae exposed to this alone. In contrast, extract from SAK containing lower concentrations of fluoride (4.25 mg kg^?1) showed no apparent adverse effects. Growth of larvae fed a mixture of SE and SAK in a feed trial of 58 days [mean body weight (BW), 6.0 mg] was about twofold higher than that of larvae fed a mixture of SE and WAK (3.2 mg) (P < 0.01). A mixture of HEY and SAK also had some dietary benefits for eel larvae, enabling them to survive for up to 58 days and to grow significantly (mean BW, 2.4 mg), compared with their initial weight (mean BW, 0.2 mg) (P < 0.001). Although additional nutritional improvements are needed, the present results suggest that combination diet HEY and SAK may be a good alternative to SE as an effective diet for eel larvae.     

Demonstration of infectious pancreatic necrosis virus strain VR-299 in Japanese eel, Anguilla japonica Temminck & Schlegel

Abstract. Six isolates of infectious pancreatic necrosis virus (IPNV) were obtained from trout or eel farms in Taiwan in 1987. By using the electrophoretic analysis of ³⁵S-methionine-labelled early viral polypeptide patterns and silver-stained ds RNA gel patterns, a comparison of these isolates with the three archetypal IPNV serotypes (AB, SP and VR-299) was made. They were all identical with VR-299, both in the RNA and early viral polypeptide patterns. From the results obtained, it was apparent that the VR-299 serotype of IPNV was just as widespread in eel as in trout in Taiwan. This is the first case of VR-299 isolation from eel.     

Haemoglobin powder as a dietary fish meal replacer in juvenile Japanese eel, Anguilla japonica (Temminck et Schlegel)

Two consecutive 6-week feeding trials were conducted to determine the amount of haemoglobin powder (BM) that could replace fish meal (FM) in juvenile Japanese eel Anguilla japonica (Temminck et Schlegel) diets. Fish were fed 50% crude protein diets in which each of ten isonitrogenous diets was formulated to contain white fish meal and/or blood meal as the dietary protein source to replace FM by BM as follows: Diet 1 (control), 0% BM; diet 2,12.5% BM; diet 3,25% BM; diet 4, 50% BM; diet 5, 75% BM; diet6,100%BM;diet7,25%BM + 3 Essential Amino Acids (EAA); diet 8, 50% BM + 3 EAA; diet 9, 75% BM + 3 EAA; diet 10, 100 BM + 3 EAA. In the first 6-week period, the results were not consistent with the treatments, and poor adaptation of the fish to the experimental diets and conditions was observed. In the second 6-week period, weight gain, specific growth rate, protein efficiency ratio and protein productive value offish fed diets 2, 3, 4, 7, 8 and 9 were not significantly different from those of fish fed the control diet (P > 0.05). However, feed conversion ratios offish fed diets 6 and 10 were lower than that offish fed the control diet (P < 0.05). These results demonstrate that FM can be replaced by BM up to 50% without supplementation of three EAA, and up to 75% with three EAA supplementation in juvenile Japanese eel diets.     

Gill filament necrosis in farmed Japanese eels, Anguilla japonica (Temminck & Schlegel), infected with Herpesvirus anguillae

A herpesviral gill disease accompanied by mass mortality occurred in Japanese eels, Anguilla japonica (Temminck & Schlegel), reared in warm water ponds from 1993 to 1995. Diseased fish displayed marked haemorrhage and congestion within gill filaments and destruction at the tips of affected filaments with necrosis and inflammation in the central connective tissue and the central sinus. Electron microscopy revealed herpesvirus particles in infected fibrocytes within the filamental connective tissue. The isolate was identified as Herpesvirus anguillae by a neutralization test. Infectivity experiments with the isolates revealed that the virus was pathogenic.     

Cell-mediated immunity of the eel, Anguilla japonica (Temminck & Schlegel), as measured by the migration inhibition test

Abstract. Sensitized lymphocytes isolated from the immunized eel, Anguilla japonica , upon interaction with the antigens in vitro , elaborated into the medium a soluble factor which was capable of inhibiting the migration of peritoneal exudate cells harvested from guinea pigs. Production of migration inhibition factor (MIF) was greatest to the specific antigen although there was also some nonspecific release. The elaboration of MIF from the sensitized lymphocytes occurred from the third day after immunization of eels and was detectable until day 106. The inhibitory activity from the blood appeared to persist over a longer period compared to that from the spleen and the anterior kidney. The inhibitory activity, however, was diminished by starvation of the immunized eels.     

日本鳗鲡(Anguilla japonica Temminck et Schlegel)鳗苗资源变动之谜

鳗苗的年产量多少（文中所指为白仔鳗，下同）波动很大，极大的影响着养鳗业。鳗苗资源的变动一直是养鳗业面临的一个严峻的问题。其资源究竟以何种机制发生变动？现就12年来对鳗苗资源动向的调查所获的情况作一概述。     

Humoral immune response of Japanese eel, Anguilla japonica Temminck & Schlegel, to Pleistophora anguillarum Hoshina, 1951 (Microspora)

A humoral response of Japanese eel, Anguilla japonica Temminck & Schlegel, to the microsporean Pleistophora anguillarum Hoshina was demonstrated using immunoblotting and an enzyme-linked immunosorbent assay (ELISA). Japanese eel immunoglobulin was purified by affinity chromatography. The immunoglobulin was composed of 25-kDa light chains and 72-kDa heavy chains. The ELISA values of P. anguillarum antibodies in naturally infected fish sera were significantly higher than those of clinically healthy fish. Spore proteins from the microsporean were separated by electrophoresis and subjected to analysis by Western blot. Sera from naturally infected fish showed different reaction patterns to the spore proteins. While the sera randomly selected from naturally infected eels all showed a significant positive reaction to P. anguillarum antigens, the mucus from only three out of the nine infected eels reacted positively in the ELISA test. Subsequent analyses indicated that there was no significant difference in the amount of mucus immunoglobulin among the tested eels. Therefore, the generally lower ELISA values of mucosal anti-P. anguillarum antibodies from the infected eels tested were evidently not caused by a lack of immunoglobulin per se, but seem to be the result of a lack of anti-P. anguillarum antibodies in the mucus and/or a lower affinity in the anti-P. anguillarum antibodies that were present.     

Isolation and characterization of bacteria causing mortality in early stage larvae of captive‐bred Japanese eels (Anguilla japonica Temminck & Schlegel)

Bacteria with lethal effect on eel larvae were isolated from moribund captive‐bred eel larvae and their 16S rRNA gene sequences were analysed. Nucleotide sequence of 10 isolates showed highest similarity with Lacinutrix algicola, Crocinitomix catalasitica and Pseudoalteromonas rubra. Age‐dependent changes in the susceptibility of eel larvae were observed in response to challenge by a highly lethal isolate. Compared with 10 and 11 days after hatching (DAH), larval susceptibility to the highly lethal isolate was lower at 18 DAH. We found that the bacterial isolates have lethal effect on the captive‐bred eel larvae, especially at the early developmental stage. These results will be useful to establish appropriate culture practices for eel larvae that will improve the success of mass production of glass eels for aquaculture.     

Protective properties of egg yolk IgY containing anti-Edwardsiella tarda antibody against paracolo disease in the Japanese eel, Anguilla japonica Temminck & Schlegel

Abstract. Immunized hens are known to contain a high level of immunoglobulin Y (IgY) in their egg yolk. In this study, the present authors obtained anti- Edwardsiella tarda IgY (containing 20% specific IgY, agglutination titre, 1:128) from hens vaccinated by injection with formalin-killed bacterin. The IgY was stable against eel digestive factors, and therefore, was orally administered with viable E. tarda to the Japanese eels and the efficacy of protection against E. tarda infection was evaluated. Orally administered IgY at a dose of 400mg fish^-1 cleared E. tarda inoculated simultaneously at 10^5–6 CFU fish^-1 from the intestine within 24h. Moreover, orally administered IgY at doses of 200 and 400mg fish^-1 inhibited the penetration of E. tarda inoculated simultaneously at 10^4–6 CFU fish^-1 into the liver and kidney via the damaged intestine. The fish orally administered with IgY showed reduced mortality. These results suggest that egg yolk containing anti- E. tarda IgY is effective in preventing edwardsiellosis.     

The roles of bone and muscle as phosphorus reservoirs during the sexual maturation of female Japanese eels, Anguilla japonica Temminck and Schlegel (Anguilliformes)

We sought to determine the organs that function as phosphorus (P) reservoirs for supplying this mineral to the ovary during the sexual maturation of the Japanese eel, Anguilla japonica (Anguilliformes). The P content of the ovary increased markedly during sexual maturation despite the lack of an extrinsic P supply (e.g., diet). This phenomenon suggests that the P that accumulated in the ovary was derived from other parts of the eel. In addition, the P content of bone dramatically decreased during maturation, as did the total bone weight. This result suggests that resorption of bone occurred during the sexual maturation of the eel. Whereas the P content and mass of muscle markedly decreased during maturation, the Ca and P contents of the skin (which was scaled) did not. The total (whole-fish) P content was constant throughout maturation. Together, our results suggest that almost all of the P lost from bone and muscle is incorporated in the ovary and that bone and muscle function as P reservoirs during the sexual maturation of the Japanese eel. This revised version was published online in August 2006 with corrections to the Cover Date.     

Studies on epizootic iridovirus infection among red sea bream,Pagrus major (Temminck & Schlegel), cultured in Taiwan

Since 1993, an epizootic viral disease has occurred in net-cage cultured red sea bream, Pagrus major (Temminck & Schlegel), in Peng-hu Island located on the south-western coast of Taiwan. The diseased fish exhibited abnormal swimming and were lethargic, but few visible external signs were observed. The cumulative mortality because of the disease sometimes reached 50-90% over 2 months. Histopathogical studies of the affected fish showed enlarged basophilic cells in the gill, kidney, heart, liver and spleen. These necrotic cells were Feulgen-positive and stained blue using Giemsa. Transmission electron microscopy revealed icosahedral virions in the cytoplasm of the necrotic cells. The viral particles consisted of a central nucleocapsid (75-80 nm) and envelope, and were 120-150 nm in diameter. These results suggest that the virus belongs to the Iridoviridae. Using polymerase chain reaction (PCR), approximately 570 bp fragments were produced from the viral DNA using as a template 1-F and 1-R primers derived from red seabream iridovirus (RSIV) from red sea bream in Japan. Similar results were also obtained using nested-PCR with different primer sets (1-F, 2-R and 2-F, 1-R). Although the size and some features of epizootics of this virus differed from RSIV in Japan, it shows close genetic affinities with the latter and it is suggested that RSIV has been introduced to Taiwan.