Development of cheap and quick diagnostic methods using muzzle skin of rabid dogs

• Project/Area Number: 26450410
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: Kitasato University
• Principal Investigator: PARK ChunHo 北里大学, 獣医学部, 准教授 (50383550)
• Co-Investigator(Renkei-kenkyūsha): Inoue Satoshi 国立感染症研究所, 獣医科学部, 室長 (90213157) ; Yamada Kentaro 大分大学, 医学部, 准教授 (70458280)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 狂犬病 / フィリピン / 犬 / 脳 / 洞毛（触毛） / 病理診断法 / メルケル細胞 / 人獣共通感染症 / 鼻口部洞毛（触毛） / 病理学 / 皮膚 / 唾液腺 / 中枢神経系

Outline of Final Research Achievements

The direct fluorescent antibody test on fresh brain tissues is the gold standard for rabies virus antigen detection in dogs. However, this method is laborious and holds a high risk of virus exposure for the experimenter. In mammals, the tactile hair is a specialized touch organ that is abundant in the muzzle skin. Each tactile hair is equipped with more than 2,000 sensory nerve endings. Therefore, this organ is expected to serve as an alternative postmortem diagnostic material. In the present study, muzzle skins were obtained from rabid dogs and the target cells and localization of rabies virus antigen was defined. Virus antigen was clearly detected in a part of the outer root sheath at the level of the ring sinus of the tactile hairs, and the majority of cells were Merkel cell. Our results suggest that Merkel cells in the tactile hairs of the muzzle skin are a target for virus replication and could serve as a useful specimen source for diagnosis of rabies.

Research Products

• [Int'l Joint Research] 熱帯医学研究所 (RITM）(フィリピン)
• [Int'l Joint Research] 台湾・家畜衛生試験所 (AHRI)(台湾)
• [Int'l Joint Research] モンゴル・獣医学研究所(モンゴル)
• [Int'l Joint Research] Veterinary Research Department/RITM/Department of Health(Philippines)
• [Journal Article] Pathological lesions in the central nervous system and peripheral tissues of ddY mice with street rabies virus (1088 strain) (2017)
  • Author(s): Kimitsuki K, Yamada K, Shiwa N, Inoue S, Nishizono A, Park CH.
  • Journal Title: J. Vet. Med. Sci. Volume: 79(6)
  • NAID: 130005695725
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Localization of the rabies virus antigen in Merkel cells in the follicle-sinus complexes of muzzle skins of rabid dogs (2016)
  • Author(s): Shimatsu T, Shinozaki H, Kimitsuki K, Shiwa N, Manalo DL, Perez RC, Dilig JE, Yamada K, Boonsriroj H, Inoue S, Park CH.
  • Journal Title: J. Virol. Methods Volume: 237(40) Pages: 40-46
  • Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] A pathological study of the salivary glands of rabid dogs in the Philippines. (2016)
  • Author(s): Boonsriroj H, Manalo DL, Kimitsuki K, Sihmatsu T, Shiwa N, Shinozaki H, Takahashi Y, Tanaka N, Inoue S, Park CH.
  • Journal Title: J. Vet. Med. Sci. Volume: 78 Pages: 35-42
  • NAID: 130005121596
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Survey of francisella tularensis among wild animals in the endemic areas in Japan (2016)
  • Author(s): Hotta A, Tanabayashi K, Fujita O, Shindo J, Park CH, Kudo N, Hatai H, Oyamada T, Yamamoto Y, Takano A, Kawabata H, Sharma N, Uda A, Yamada A, Morikawa S.
  • Journal Title: Jpn. J. Infect. Dis. Volume: inpress
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ザーネン種山羊の脳脊髄糸状虫症の１例 (2016)
  • Author(s): 中本悠香、志和 希、 島津太一、君付和範、 富岡美千子,、片山芳也、筏井宏実、朴 天鎬
  • Journal Title: 日獣会誌 Volume: 69 Pages: 36-40
  • NAID: 130005126759
  • Peer Reviewed / Open Access
• [Journal Article] The co-existence of a myxoid liposarcoma and leiomyoma in the same ovarian mass of a dog (2016)
  • Author(s): Shiwa N, Boonsriroj H, Kimitsuki K, Shimatsu T, Park CH.
  • Journal Title: J. Vet. Med. Sci. Volume: 78 Pages: 467-471
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Congenital lobar emphysema concurrent with pneumothorax and pneumomediastinum in a dog (2016)
  • Author(s): Yun S, Lee H, Lim J, Lee K, Jang K, Shiwa N, Boonsriroj H, Kimitsuki K, Park CH, Kwon Y.
  • Journal Title: J. Vet. Med. Sci. Volume: in press
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Association between RABV G proteins transported from the perinuclear space to cell surface membrane and N-glycosylation of the sequon at Asn204 (2015)
  • Author(s): Hamamoto N, Uda A, Tobiume M, Park CH, Noguchi A, Kaku Y, Okutani A, Morikawa S, Inoue S
  • Journal Title: JJID Volume: in press
  • NAID: 130005098925
  • Peer Reviewed / Open Access
• [Presentation] フィリピンの熱帯医学研究所で狂犬病陽性と診断された発症犬の神経病理学的研究 (2017)
  • Author(s): 志和希、川本尚未、君付和範、Daria Llenaresas Manalo、井上智、 朴天鎬
  • Organizer: 第4回日本獣医病理学専門家協会学術集会
  • Place of Presentation: タワーホール船堀（東京江戸川区）
  • Year and Date: 2017-03-30
• [Presentation] 狂犬病発症犬の鼻口部洞毛組織の死後診断的有用性の再確認と洞毛組織における病理学的所見 (2016)
  • Author(s): 中島千景, Daria Llenaresas Manalo, 君付和範, 川本尚未, 須見康代, 山岸光葉, 篠崎春美, 島津太一, 志和 希, 山田健太郎, 井上智, 朴天鎬
  • Organizer: 第159回日本獣医学会学術集会
  • Place of Presentation: 日本大学(神奈川県藤沢市）
  • Year and Date: 2016-10-04
• [Presentation] Merkel cells in the follicle-sinus complexes of muzzle skins are useful for postmortem diagnosis of rabid dogs (2016)
  • Author(s): Park CH, Shimatsu T, Kimitsuki K, Shiwa N, Manalo DL, Inoue S
  • Organizer: The Korean Society of Veterinary Science
  • Place of Presentation: Korea, Seoul
  • Year and Date: 2016-04-29
  • Int'l Joint Research
• [Presentation] Pathological study on the taste buds and von Ebner’s glands of rabid dogs in the Philippines (2015)
  • Author(s): Shiwa N, Sumi Y, Shimatsu T, Kimitsuki K, Boonsriroj H, Manalo DL, Inoue S, Park CH.
  • Organizer: 7th Asian Society of Veterinary Pathology (ASVP)
  • Place of Presentation: University of the Philippines（Philippines）
  • Year and Date: 2015-11-07
  • Int'l Joint Research
• [Presentation] A pathological study of the salivary glands of rabid dogs in the Philippines (2015)
  • Author(s): Kimitsuki K, Boonsriroj H, Manalo DL, Shimatsu T, Shiwa N, Takahashi Y, Tanaka N, Inoue S, Park CH.
  • Organizer: 7th Asian Society of Veterinary Pathology (ASVP)
  • Place of Presentation: University of the Philippines（Philippines）
  • Year and Date: 2015-11-07
  • Int'l Joint Research
• [Presentation] The utility of muzzle skin of rabid dogs as antemortem and postmortem diagnosis (2015)
  • Author(s): Shimatsu T, Kawamoto N, Shiwa N, Kimitsuki K, Boonsriroj H, Manalo DL, Shinozaki H, Takahashi Y, Tanaka N, Inoue S, Park CＨ.
  • Organizer: 7th Asian Society of Veterinary Pathology (ASVP)
  • Place of Presentation: University of the Philippines（Philippines）
  • Year and Date: 2015-11-07
  • Int'l Joint Research
• [Presentation] Congenita lobar emphysema concurrent with pneumothorax and pneumomediastinum in a dog (2015)
  • Author(s): Kim E, Yun SH, Lim J, Jang KH, Park CH, Kwon YS
  • Organizer: 7th Asian Society of Veterinary Pathology (ASVP)
  • Place of Presentation: University of the Philippines（Philippines）
  • Year and Date: 2015-11-07
  • Int'l Joint Research
• [Presentation] 直接迅速免疫組織化学（DRIT）法を利用したフィリピンにおける狂犬病検査法の検討 (2015)
  • Author(s): Manalo L. Daria, Bazartseren Boldbaatar, 朴 天鎬, 井上 智
  • Organizer: 第158回日本獣医学会学術集会
  • Place of Presentation: 北里大学十和田キャンパス（青森県十和田市）
  • Year and Date: 2015-09-07
• [Presentation] 動物由来感染症を想定した解剖施設紹介と狂犬病発症犬の病理組織学 (2015)
  • Author(s): 朴天鎬
  • Organizer: 平成27年度狂犬病予防業務地方ブロック技術研修会
  • Place of Presentation: 岩手県環境保健研究センター（岩手県盛岡市）
  • Year and Date: 2015-06-22
  • Invited
• [Presentation] Pathological Study on the Central Nerve System and Peripheral Tissues of ddY Mice Intramuscularly Infected with Street Rabies Virus (1088 Strain) (2015)
  • Author(s): Park, CH, Yamada, K, Kimitsuki, K, Yanogawa, S, Hassadin, B, Inoue, S, Nishizono, A.
  • Organizer: 17th International Conference on Emerging Infectious Diseases (EID)
  • Place of Presentation: 台湾中央研究院ACADEMICA SINICA（台北市）
  • Year and Date: 2015-01-26 – 2015-01-29
• [Presentation] Pathological Studies on the Salivary Glands of Rabid Dogs (2015)
  • Author(s): Hassadin, B, Takahashi, Y, Tanaka, N, Shinozaki, H, Kimitsuki, K, Daria, L, Manalo, Inoue, S, Park, CH.
  • Organizer: 7th International Conference on Emerging Infectious Diseases (EID)
  • Place of Presentation: 台湾中央研究院ACADEMICA SINICA（台北市）
  • Year and Date: 2015-01-26 – 2015-01-29
• [Presentation] Different intracellular localization of G protein between Kyoto strain (street virus) and CVS-26 strain (fixed virus) was associated with N-glycosylation of G protein at the site of 204 (2015)
  • Author(s): Hamamoto N, Uda A, Noguchi A, Okutani A, Kaku Y, Park CH, Morikawa S, Inoue S.
  • Organizer: 17th International Conference on Emerging Infectious Diseases (EID)
  • Place of Presentation: 台湾中央研究院ACADEMICA SINICA（台北市）
  • Year and Date: 2015-01-26 – 2015-01-29
• [Presentation] 狂犬病発病犬の口唇皮膚におけるウイルス抗原の局在と診断的有用性 (2014)
  • Author(s): 篠崎春美、高橋友里花、矢野川祥子、君付和範、Hassadin Boonsriroj、Daria Llenaresas Manalo、井上智、朴 天鎬
  • Organizer: 第156回日本獣医学会学術集会
  • Place of Presentation: 北海道大学（札幌）
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] 狂犬病発病犬の唾液腺に関する病理学的研究 (2014)
  • Author(s): 高橋友里花、篠崎春美、矢野川祥子、君付和範、Hassadin Boonsriroj、Daria Llenaresas Manalo、井上智、朴天鎬
  • Organizer: 第156回日本獣医学会学術集会
  • Place of Presentation: 北海道大学（札幌）
  • Year and Date: 2014-09-09 – 2014-09-12