{"created":"2023-06-19T09:48:56.254802+00:00","id":5324,"links":{},"metadata":{"_buckets":{"deposit":"cdf319cf-232c-4f28-9f12-f4b2b4257fcd"},"_deposit":{"created_by":3,"id":"5324","owners":[3],"pid":{"revision_id":0,"type":"depid","value":"5324"},"status":"published"},"_oai":{"id":"oai:fukuyama-u.repo.nii.ac.jp:00005324","sets":["502:506:514:516"]},"author_link":["26269","26268","26272","26264","26270","26265","26267","26271","26266"],"item_1_biblio_info_14":{"attribute_name":"書誌情報","attribute_value_mlt":[{"bibliographicIssueDates":{"bibliographicIssueDate":"1999-12","bibliographicIssueDateType":"Issued"},"bibliographicPageEnd":"69","bibliographicPageStart":"1","bibliographicVolumeNumber":"10","bibliographic_titles":[{"bibliographic_title":"福山大学内海生物資源研究所報告"}]}]},"item_1_creator_6":{"attribute_name":"著者名(日)","attribute_type":"creator","attribute_value_mlt":[{"creatorNames":[{"creatorName":"芦田, 貴行"}],"nameIdentifiers":[{"nameIdentifier":"26264","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"沖増, 英治"}],"nameIdentifiers":[{"nameIdentifier":"26265","nameIdentifierScheme":"WEKO"}]},{"creatorNames":[{"creatorName":"雨村, 明倫"}],"nameIdentifiers":[{"nameIdentifier":"26266","nameIdentifierScheme":"WEKO"}]}]},"item_1_description_1":{"attribute_name":"ページ属性","attribute_value_mlt":[{"subitem_description":"P(論文)","subitem_description_type":"Other"}]},"item_1_description_11":{"attribute_name":"抄録(日)","attribute_value_mlt":[{"subitem_description":"主要海産養殖魚であるヒラメのエドワジエラ症は,1982年頃から発生するようになり、その原因菌としてEdwardsiella tardaが分離された。その後,本症は初夏から晩秋にかけての高水温期に魚令を問わず発生するようになり,1995年以降には多発するようになった。現在では,ヒラメ養殖において最も被害の大きな疾病となっている。本症に対する治療薬としては,塩酸オキシテトラサイクリンが用いられているが,薬剤の多用によって耐性菌が出現するようになり,大きな問題となっている。したがって,より効果的な予防・治療法を確立することが重要な課題となってきた。そこで,本章では前節においてブリの腸球菌症に対して効果が認められたquillaja saponinと,魚類の免疫機構を活性化する効果があるとされているβ-1,3-glucan (curdlan)の2種類の生理活性物質,およびワクチンをヒラメに投与して,エドワジエラ症に対する投与効果を検討した。これら2種の生理活性物質うち,quillaja saponinのヒラメのエドワジエラ症に対する防御効果は,多少の延命効果が認められるものの有効性には大きな期待ができないことが明らかになった。Curdlanについては,ヒラメの生体防御能に与える投与期間および投与法の影響を白血球の貧食能とPMA刺激によるO_2^-産生率から調べて検討した。その結果,curdlanを連続投与したヒラメ白血球の貧食能は,投与開始後2週間までは対照区に比較して上昇したが,3週間以上投与すると活性が減少して投与開始前の水準まで低下した。また,curdlan投与はO_2^-産生に影響しなかった。しかし,curdlanを3週間投与したヒラメのPMNに,in virtoでcurdlanを添加したところ,O_2^-産生率が上昇した。いっぽう,4か月間curdlanを隔日投与したヒラメ白血球の貧食能は,対照区に比較して高くなった。しかし,その活性は短期間の連続投与に比較して低下した。これらのことから,curdlan投与がヒラメの非特異的免疫機構に与える効果は,その投与法と投与期間によって影響されることが明らかになった。そこで,白血球貧食能に対する活性化効果が顕著に認められたcurdlanを,2週間連続投与したのちに,E. tardaで実験感染を行って有効性を評価したところ,curdlan投与による顕著な延命効果は認められなかった。そこで,E. tardaのホルマリン不活化菌体をワクチンとして経口投与し,特異的な防御機構を活性化してワクチンの効果を検討した。その結果,ワクチンの単独投与では血液中抗体価の上昇が認められ,有効率も生理活性物質を単独投与した場合に比較して高い値となったが,顕著な延命効果は認められなかった。したがって、次にワクチンとquillaja saponinおよびcurdlanの2種類の生理活性物質の併用投与を検討した。ワクチンと生理活性物質を併用投与したヒラメでは,ワクチンを単独投与した場合に比較して抗体価と負食能が上昇した。したがって、quillaja saponinおよびcurdlanにはアジュバント効果があることが示唆された。また,ワクチンとこれら2種の生理活性物質を併用投与すると,E. tarda実験感染に対する有効率が高められた。このように,ワクチンや生理活性物質の投与は単独では十分な有効性は得られなかったが,併用すると,相乗効果が認められることが明らかになった。しかし,これらの併用投与は白血球のO_2^-産生には影響しなかった。ヒラメにワクチンとquillaja saponinとcurdlanの2種の生理活性物質を併用投与し,感染前後の腸管,腎臓および脾臓のhsp60の変化を調べ,併用投与が,hsp60の発現にどのような影響を及ぼすかについて検討した。その結果,腸管と腎臓および脾臓とではhsp60の誘導様式に差異が認められた。すなわち,腸管のhsp60はワクチンと生理活性物質の併用投与区では,対照区に比較して感染前に上昇する傾向が認められた。しかし,腎臓と脾臓においては併用投与しても,腸管のような投与後のhsp60レベルの上昇は認められなかった。腸管は病原体などの侵入経路であることから,粘膜組織に特有な粘膜免疫機構が発達しているとされている。粘膜免疫機構は,腸管上皮細胞間に存在するリンパ球であるγ,δ-T細胞がhsp60を認識して,活性化されると考えられている。これらのことから,ワクチンと生理活性物質との併用投与区の腸管では,投与後のhsp60レベルが上昇して粘膜免疫機構が活性化されているのではないかと推察された。ワクチンと生理活性物質とを併用投与すると,腸管のhsp60レベルが上昇し,粘膜免疫系の活性化を促進して延命効果が高くなるのではないかと思われた。いっぽう,腎臓では対照区の感染後のhsp60レベルが上昇した。腎臓のhsp60は前章のブリの腸球菌症の試験感染においても感染前では低く,感染後に高くなった。また,脾臓ではワクチンとcurdlanの併用投与区および対照区の感染後のhsp60レベルが,高い値を示した。したがって,腎臓および脾臓のhsp60は粘膜免疫機構の発達した腸管とは異なり,炎症組織の保護と修復に関与するシャペロニンとして機能していると思われた。腎臓および脾臓のhsp60レベルが高い病魚では,炎症が沈静化されておらず,細菌感染によるストレスが高い状態にあるのではないかと推察された。これに対して,ワクチンと2種の生理活性物質の併用投与区では,感染前と感染後の生残魚の腎臓と脾臓のhsp60レベルに有意差は認められなかった。したがって,併用投与区の病魚の腎臓および脾臓の感染によるストレスは,低いと思われた。これらのことから,ワクチンと生理活性物質との併用投与では,感染後のヒラメにおいて症状が発現しなかったか,あるいは治癒傾向にあったと推察された。ワクチンと生理活性物質の経口投与は,単独では十分な防御効果は得られないが,併用によって血中抗体価,白血球の貧食能および試験感染に対する有効率も高くなった。また,感染後の病魚では症状が発現しないか,発現してもそれらは治癒傾向にあったと推察された。したがって,ワクチンと生理活性物質を併用した経口投与法は,ヒラメの本症に対する有効な防御手段となるのではないかと思われた。","subitem_description_type":"Other"}]},"item_1_description_12":{"attribute_name":"抄録(英)","attribute_value_mlt":[{"subitem_description":"Yellowtail, Seriola quinqueradiata, and Japanese flounder, Paralichthys olivaceus, are popular cultured marine fish in Japan. The enterococcicosis of yellowtail and edwardsiellosis of Japanese flounder are serious problems for fish culture. Antibiotics are effective for treating these bacterial diseases, but the easy use of these chemicals has led to the appearance of drug-resistant bacteria and the persistence of the drugs in the fish. In this study, more effective techniques for preventing these diseases involving the use of immunostimulants and a vaccine, which will prevent the appearance of antibiotic-resistant bacteria and harm to our health, were developed. First, the effects of quillaja and beet saponins on the resistance against enterococcicosis of yellowtail was examined. The survival of infected fish was monitored for 9 days after immersion challenge. The results showed that all fish in the control (without saponin) for the challenge groups had died by day 6 after the challenge, whereas the rate of survival of fish in the saponin-fed groups were 40%. Thus, the feeding of saponins was found to elevate the survival rate after bacterial challenge. In these experiments the amount of lipid peroxide (LPO) in the liver increased after the enterococcicosis, however, those in the liver of saponin-fed fish did not increase after the infection. On the other hand, the activities of superoxide clismutase (SOD) and glutathioneperoxidases (GPxs), which are active oxygen-scavenging enzymes, increased in the kidney and spleen of yellowtail after the challenge, and saponin feeding resulted in progressive increases in the activities following the challenge. The activation of these enzymes was thought to reduce the oxidative stress caused by the enterococcicosis. Infection and oxidative stress induce a serious stress response to organisms, resulting in the acquisition of stress resistance to the organisms. Chaperone proteins, hsps, whose function is to prevent.the improper aggregation or association of partially folded proteins, prevent the denaturation of proteins due to various forms of stress such as heat and infection. In the kidney and spleen, hsp60 and hsp32 were produced continuously at high levels in the saponin-fed fish. These results showed that saponins seem to promote hsp induction. The promotion of active oxygenscavenging enzymes and hsp induction on pre-feeding of saponins may help to reduce the bacterial infection stress more effectively, and the reduced infection stress may increase the survival rate after enterococcicosis of yellowtail. Next, the effect of quillaja saponin feeding on edwardsiellosis of Japanese flounder was studied, however, the effect was low. Therefore, we investigated the effect of bacterial β-1,3-glucan (curcllan) on this disease, because some β-glucans are known to increase the non-specific immune response of a host. In a short period (1, 2, 3 and 4 weeks) feeding test, the phagocytic activity of Japanese flounder increased until 2 weeks with everyday feeding of curdlan, and the superoxide generation in fish fed curdlan every clay for 3 weeks was enhanced from 10 to 20% with the low concentrations of curdlan (in vitro). When fish were fed every 2 days for 4 months (long period feeding test) their phagocytic activity increased, however, levels were lower than in the short period feeding test, whereas the superoxide generation did not increase. Thus, the feeding period and feeding mode influenced the effect of curdlan. Then, the effect of a vaccine (formalin-killed Edwardsiella tarda) on activation of the specific immune defense mechanism in Japanese flounder was studied. The agglutination titer of Japanese flounder fed diets containing the vaccine was higher than that in the vaccine-unsupplemented diet group, however, the survival rate of the vaccine group (30%) was not notably higher than that in the curdlan or quillaja saponin-fed fish, or the control (without an immunostimulant) after immersion challenge. For effective treatment, we examined the combined effects of these immunostimulants and the vaccine. When the vaccine was fed with curdlan and quillaja saponin, the survival rates of E. tarda-infected fish (42%) increased significantly compared to the control group. In these fish, the phagocytic activity of leucocytes and the antibody titers in serum were higher than in vaccine only fed fish, however, the combinations of the immunostimulants and vaccine were not effective for the superoxide generation by leucocytes. Finally, the influence of oral administration of quillaja saponin, curdlan and vaccine on the hsp60 expression levels in the intestine, kidney and spleen of Japanese flounder was examined. In the immunostimulants and vaccine-fed fish, the hsp60 expression pattern in the intestine was found to be different from those in the kidney and spleen, i.e. the hsp60 level in the intestine increased more highly compared to the control (no feeding of an immunostimulants or the vaccine), but did not increase significantly with the levels in the kidney and spleen. It was suggested that the surfaces of mucosal tissues such as the intestine, and respiratory and urogenital tracts develop a special immune defense system: M cells which exist in the intestinal epithelium transport antigens for efficient endocytosis and transcytosis; the antigens transported by M cells are processed and presented by macrophages, and then the macrophages activate γ,δ-T cells in the intraepithelial pocket. The activation of the mucosal immune defense system is thought to be caused by hsps recognized by γ,δ-T cells. Thus, hsp60 seems to regulate the T cell response against this disease as an important function in the activation of the mucosal immune system, and the immunostimulants and vaccine we used seem to stimulate the expression of hsp60. However, after infection the hsp60 level in the intestine decreased. The reason for this is that cells of the intestine were disrupted or the mucosal immune system was damaged by edwardsiellosis. On the other hand, the hsp60 levels in the kidney and spleen of the control group increased after infection. These results showed previously that hsp60 increased in the kidney of yellowtail on enterococcicosis. The abnormal expression of such stress proteins has been widely observed in a number of infection conditions including oxidant injury, ischemia and inflammation. The hsp60 induced in these organs may have protected them from the oxidative stress caused by bacterial infection. Thus, it seems that hsp 60 induction is an important event induced by bacterial infection stress in the kidney or other organs. As the hsp60 level in the intestine decreased with edwardsiellosis, probably through damage to the mucosal immune system, as mentioned above, we will be able to determine the extent of infection stress in fish by measuring the hsp60 levels in organs. In conclusion, the feeding of saponins elevated the hsp60 levels in some organs of yellowtail, which led to a higher survival rate of yellowtail after s enterococcicosis. In Japanese flounder, the single feeding of saponin, curdlan or the vaccine was not effective against edwardsiellosis. However, the combination of these immunostimulants and vaccine raised the agglutination antibody titers in serum and triggered the phagocytic activity of leucocytes, which enhanced the ultimate survival rate of fish after infection as a result. The method of oral administration of immunostimulants and vaccine used here was better than intraperitoneal injection, spraying and immersion, since with this method a mass of fish is immunized with very little handling stress.","subitem_description_type":"Other"}]},"item_1_full_name_7":{"attribute_name":"著者名よみ","attribute_value_mlt":[{"nameIdentifiers":[{"nameIdentifier":"26267","nameIdentifierScheme":"WEKO"}],"names":[{"name":"アシダ, タカユキ"}]},{"nameIdentifiers":[{"nameIdentifier":"26268","nameIdentifierScheme":"WEKO"}],"names":[{"name":"オキマス, エイジ"}]},{"nameIdentifiers":[{"nameIdentifier":"26269","nameIdentifierScheme":"WEKO"}],"names":[{"name":"アメムラ, アキノリ"}]}]},"item_1_full_name_8":{"attribute_name":"著者名(英)","attribute_value_mlt":[{"nameIdentifiers":[{"nameIdentifier":"26270","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Ashida, Takayuki","nameLang":"en"}]},{"nameIdentifiers":[{"nameIdentifier":"26271","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Okimasu, Eiji","nameLang":"en"}]},{"nameIdentifiers":[{"nameIdentifier":"26272","nameIdentifierScheme":"WEKO"}],"names":[{"name":"Amemura, Akinori","nameLang":"en"}]}]},"item_1_source_id_13":{"attribute_name":"雑誌書誌ID","attribute_value_mlt":[{"subitem_source_identifier":"AA1157543X","subitem_source_identifier_type":"NCID"}]},"item_1_text_10":{"attribute_name":"著者所属(英)","attribute_value_mlt":[{"subitem_text_language":"en","subitem_text_value":"Doctoral Programsin Life Scienceand Biotechnology, Graduate School of Engineering, Fukuyama University"},{"subitem_text_language":"en","subitem_text_value":"Department of Marine Biotechnology, Fukuyama University"},{"subitem_text_language":"en","subitem_text_value":"Department of Marine Biotechnology, Fukuyama University"}]},"item_1_text_2":{"attribute_name":"記事種別(日)","attribute_value_mlt":[{"subitem_text_value":"原著論文"}]},"item_1_text_3":{"attribute_name":"記事種別(英)","attribute_value_mlt":[{"subitem_text_language":"en","subitem_text_value":"Original"}]},"item_1_text_9":{"attribute_name":"著者所属(日)","attribute_value_mlt":[{"subitem_text_value":"福山大学大学院生命工学専攻博士後期課程"},{"subitem_text_value":"福山大学海洋生物工学科"},{"subitem_text_value":"福山大学海洋生物工学科"}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"1999-12-01"}],"displaytype":"detail","filename":"KJ00005781881.pdf","filesize":[{"value":"4.2 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"url":"https://fukuyama-u.repo.nii.ac.jp/record/5324/files/KJ00005781881.pdf"},"version_id":"7d981b3f-dc9e-48d5-85d4-5b34e623b858"}]},"item_keyword":{"attribute_name":"キーワード","attribute_value_mlt":[{"subitem_subject":"yellowtail","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"enterococcicosis","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"immunostimulant","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"active oxygens","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"heat shock protein","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"Japanese flounder","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"eclwarclsiellosis","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"oral vaccine","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"agglutination titer","subitem_subject_language":"en","subitem_subject_scheme":"Other"},{"subitem_subject":"phagocytosis","subitem_subject_language":"en","subitem_subject_scheme":"Other"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"jpn"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"departmental bulletin paper","resourceuri":"http://purl.org/coar/resource_type/c_6501"}]},"item_title":"ブリの腸球菌症とヒラメのエドワジエラ症の防御に関する研究","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"ブリの腸球菌症とヒラメのエドワジエラ症の防御に関する研究"},{"subitem_title":"Studies on Protection against Enterococcicosis of Yellowtail and Edwardsiellosis of Japanese Flounder","subitem_title_language":"en"}]},"item_type_id":"1","owner":"3","path":["516"],"pubdate":{"attribute_name":"公開日","attribute_value":"1999-12-01"},"publish_date":"1999-12-01","publish_status":"0","recid":"5324","relation_version_is_last":true,"title":["ブリの腸球菌症とヒラメのエドワジエラ症の防御に関する研究"],"weko_creator_id":"3","weko_shared_id":-1},"updated":"2023-06-19T11:26:14.198255+00:00"}