Development of a new diagnostic procedure for bovine leukemia virus infection using bovine milk exosomes

• Project/Area Number: 26660239
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Veterinary medical science
• Research Institution: Gifu University
• Principal Investigator: Inoshima Yasuo 岐阜大学, 応用生物科学部, 准教授 (20355184)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: エキソソーム / 生乳 / バルク乳 / 牛白血病 / 牛白血病ウイルス / 診断法

Outline of Final Research Achievements

We tried to establish a method for efficient isolation and purification of exosomes from bovine raw milk and to develop a new diagnostic procedure for bovine leukemia virus (BLV) infection using milk exosomes. Our study revealed that commercially available exosome-isolation kits are applicable to isolate bovine milk exosomes after removal of milk casein by sequential ultracentrifugation. BLV antigens and DNA were not detected from milk exosomes from BLV-infected young cattle subclinically. These results suggest that detectability of BLV antigens and DNA in milk exosomes are conceivably related to the copy numbers in cattle.

Research Products

• [Journal Article] Evaluating the Stability of Loop-Mediated Isothermal Amplification Reagents at Irregular Storage Temperatures for On-Site Diagnosis (2015)
  • Author(s): Inoshima, Y., Ishiguro, N.
  • Journal Title: JARMAM Volume: 26 Pages: 7-12
  • Peer Reviewed
• [Journal Article] Role of cell death in the propagation of PrPSc in immune cells (2015)
  • Author(s): Takahashi, K., Inoshima, Y., Ishiguro, N.
  • Journal Title: Archives of Virology Volume: 160 Issue: 3 Pages: 693-399
  • DOI: 10.1007/s00705-014-2320-z
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Long-term kinetics of AA amyloidosis and effects of inflammatory restimulation after disappearance of amyloid depositions in mice (2015)
  • Author(s): Muhammad, N., Murakami, T., Inoshima, Y., Ishiguro, N.
  • Journal Title: Clinical and Experimental Immunology Volume: 181 Issue: 1 Pages: 133-141
  • DOI: 10.1111/cei.12615
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Systemic AA amyloidosis as a prion-like disorder (2015)
  • Author(s): Murakami, T., Inoshima, Y., Ishiguro, N.
  • Journal Title: Virus Research Volume: 207 Pages: 76-81
  • DOI: 10.1016/j.virusres.2014.12.019
  • Peer Reviewed / Open Access
• [Journal Article] Effect of heating on the stability of amyloid A (AA) fibrils and the intra- and cross-species transmission of AA amyloidosis (2015)
  • Author(s): Ogawa, S., Murakami, T., Inoshima, Y., Ishiguro, N.
  • Journal Title: Amyloid Volume: 22 Issue: 4 Pages: 236-243
  • DOI: 10.3109/13506129.2015.1095735
  • Peer Reviewed / Open Access
• [Journal Article] 新生子豚にみられた感染後期の豚痘 (2015)
  • Author(s): 下田智彦、竹馬工、中山季大、鈴木義久、中西運悦、小畑晴美、猪島康雄
  • Journal Title: 家畜診療 Volume: 62 Pages: 167-173
  • Peer Reviewed
• [Journal Article] 徳島県で確認された離乳豚での豚痘 (2015)
  • Author(s): 井口陽香、小島久美子、鈴木幹一郎、山本由美子、尾川誠次郎、芝原友幸、猪島康雄
  • Journal Title: 日本豚病研究会報 Volume: 66 Pages: 26-29
  • NAID: 50009481709
  • Peer Reviewed / Open Access
• [Journal Article] On-site Visual Diagnosis of Parapoxvirus Infection Using a Portable Cordless Incubator (2014)
  • Author(s): Inoshima, Y., Ishiguro, N.
  • Journal Title: Analytical Sciences Volume: 30 Issue: 12 Pages: 1169-1173
  • DOI: 10.2116/analsci.30.1169
  • NAID: 130004704355
  • ISSN: 0910-6340, 1348-2246
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Up-Regulation of MUC2 Mucin Expression by Serum Amyloid A3 Protein in Mouse Colonic Epithelial Cells (2014)
  • Author(s): Shigemura, H., Ishiguro, N., Inoshima, Y.
  • Journal Title: Journal of Veterinary Medical Science Volume: 76 Issue: 7 Pages: 985-991
  • DOI: 10.1292/jvms.14-0007
  • NAID: 130003391314
  • ISSN: 0916-7250, 1347-7439
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Reconstructing the colonization history of lost wolf lineages by the analysis of the mitochondrial genome (2014)
  • Author(s): Matsumura, S., Inoshima, Y., Ishiguro, N.
  • Journal Title: Molecular Phylogenetics and Evolution Volume: 80 Pages: 105-112
  • DOI: 10.1016/j.ympev.2014.08.004
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Identification and characterization of a multispecific monoclonal antibody G2 against chicken prion protein (2014)
  • Author(s): Kamatari, Y. O., Ohta, S., Inoshima, Y., Oda, M., Maruno, T., Kobayashi, Y., Ishiguro, N.
  • Journal Title: Protein Science Volume: 23 Issue: 8 Pages: 1050-1059
  • DOI: 10.1002/pro.2491
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Surveillance of Amyloid Deposition and Bacterial Contamination in Chicken Liver from Meat Market (2014)
  • Author(s): Ishiguro N, Murakami T, Abdelazim EE, Inoshima Y.
  • Journal Title: The Journal of Poultry Science Volume: 51 Issue: 1 Pages: 104-107
  • DOI: 10.2141/jpsa.0130028
  • NAID: 130004446425
  • ISSN: 1346-7395, 1349-0486
  • Peer Reviewed / Open Access
• [Presentation] 小型鯨類における新規組織特異的損傷マーカーの探索-マイクロRNAを指標に- (2015)
  • Author(s): 瀬川太雄、小林由紀、鈴木美和、前田健、猪島康雄、遠藤智子、伊藤琢也
  • Organizer: 平成27年度日本水産学会
  • Place of Presentation: 仙台
  • Year and Date: 2015-09-22
• [Presentation] 腸管上皮での粘液ムチン発現に関与する血清アミロイドA3のアミノ酸領域の同定 (2015)
  • Author(s): 田代真奈美、猪島康雄、石黒直隆
  • Organizer: 第158回日本獣医学会
  • Place of Presentation: 十和田
  • Year and Date: 2015-09-07
• [Presentation] 疾患モデルマウスにおけるAAアミロイド症の伝播性 (2015)
  • Author(s): 前田麻友子、猪島康雄、石黒直隆
  • Organizer: 第158回日本獣医学会
  • Place of Presentation: 十和田
  • Year and Date: 2015-09-07
• [Presentation] ワクチン関連ニワトリAAアミロイド症に関する研究 (2015)
  • Author(s): 村上智亮、衣斐彼方、Naeem Muhammad、猪島康雄、御領政信、柳井徳麿、石黒直隆
  • Organizer: 第3回日本アミロイドーシス研究会
  • Place of Presentation: 東京
  • Year and Date: 2015-08-21
• [Presentation] 簡易DNA抽出とLAMP法による野外でのパラポックスウイルス感染症診断 (2015)
  • Author(s): 猪島康雄、石黒直隆
  • Organizer: 第27回臨床微生物迅速診断研究会
  • Place of Presentation: 金沢
  • Year and Date: 2015-07-04
• [Presentation] バンドウイルカにおけるアクアポリン2の選択的スプライシング産物と細胞の浸透圧耐性 (2014)
  • Author(s): 鈴木美和、伊藤琢也、瀬川太雄、岩﨑俊秀、前田健、野口慧多、猪島康雄、香山薫
  • Organizer: 平成２６年度日本水産学会
  • Place of Presentation: 福岡
  • Year and Date: 2014-09-19 – 2014-09-22
• [Presentation] パラポックスウイルス感染症の野外での迅速診断法の開発 (2014)
  • Author(s): 猪島康雄、石黒直隆
  • Organizer: 第１５７回日本獣医学会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] Long-term study of amyloid transmission within and across the animal species in murine AA amyloidosis model (2014)
  • Author(s): Muhammad, N., Murakami, T., Ogawa, S., Inoshima, Y., Ishiguro, N.
  • Organizer: 第１５７回日本獣医学会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] 加熱処理がAAアミロイドの伝播性に与える影響 (2014)
  • Author(s): 小川紗季、村上智亮、猪島康雄、石黒直隆
  • Organizer: 第１５７回日本獣医学会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] 絶滅した日本のオオカミの遺伝的系譜 (2014)
  • Author(s): 石黒直隆、猪島康雄、柳井徳麿
  • Organizer: 第１５７回日本獣医学会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] 2007年以前の国内野生動物における重症熱性血小板減少症候群ウイルス(SFTSV)に対する抗体調査 (2014)
  • Author(s): 堀田明豊、木村昌伸、坪田敏男、中村幸子、片山敦司、中下留美子、猪島康雄、鈴木道雄、今岡浩一、棚林清、藤田修、山本美江、宇田晶彦、森川茂
  • Organizer: 第１５７回日本獣医学会
  • Place of Presentation: 札幌
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] 農場でのパラポックスウイルス感染症診断を目指したコードレス恒温維持装置開発の試み (2014)
  • Author(s): 猪島康雄、石黒直隆
  • Organizer: 第２６回臨床微生物迅速診断研究会
  • Place of Presentation: 岡山
  • Year and Date: 2014-07-26
• [Presentation] 腸管上皮における血清アミロイドA3によるムチン産生の亢進 (2014)
  • Author(s): 猪島康雄、重村洋明、石黒直隆
  • Organizer: 第７９回日本インターフェロン・サイトカイン学会
  • Place of Presentation: 札幌
  • Year and Date: 2014-06-19 – 2014-06-20
• [Presentation] Long-term study of AA amyloidosis mechanism within and across the animal species (2014)
  • Author(s): Muhammad, N., Murakami, T., Inoshima, Y., Ishiguro, N.
  • Organizer: XIV International Symposium on Amyloidosis
  • Place of Presentation: Indianapolis, IN, USA
  • Year and Date: 2014-04-27 – 2014-05-01
• [Book] ポックスウイルス、アスファウイルス、イリドウイルス（コアカリ獣医微生物学） (2015)
  • Author(s): 猪島康雄
  • Total Pages: 5
  • Publisher: 文永堂出版
• [Book] 牛丘疹性口炎、偽牛痘 （獣医公衆衛生学II） (2014)
  • Author(s): 猪島康雄
  • Total Pages: 1
  • Publisher: 文永堂出版
• [Remarks] 岐阜大学 応用生物科学部 共同獣医学科 食品環境衛生学研究室
  • URL: http://www1.gifu-u.ac.jp/~naishigu/index.html
• [Remarks] 食品環境衛生学研究室
  • URL: http://www1.gifu-u.ac.jp/~naishigu/index.html