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Affiliation |
Center for Animal Disease Control |
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Associate Professor |
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Related SDGs |
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MEKATA Hirohisa
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Research Areas 【 display / non-display 】
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Life Science / Veterinary medical science
Papers 【 display / non-display 】
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Direct TaqMan assay for the detection and genotyping of bovine viral diarrhea virus types 1 and 2 Reviewed International coauthorship International journal
Shakir Ullah, Kosuke Notsu, Akatsuki Saito, Tamaki Okabayashi, Hirohisa Mekata, Norikazu Isoda, Satoshi Sekiguchi
Archives of Virology 170 ( 1 ) 1 - 10 2025.1
Language：English Publishing type：Research paper (scientific journal) Publisher：Springer Science and Business Media LLC
Abstract
Bovine viral diarrhea (BVD), caused by bovine viral diarrhea virus (BVDV), has a significant economic impact on affected farms worldwide. For effective disease control, it is crucial to select an appropriate vaccine based on the specific genotype of BVDV. Therefore, developing a rapid and reliable assay to detect and genotype BVDV is imperative for controlling the spread of disease. In this study, we developed a TaqMan assay to detect and genotype BVDV types 1 and 2 directly in bovine serum without extraction of RNA. The direct BVDV TaqMan assay effectively detected both BVDV1 and BVDV2 with confirmed specificity and showed no cross-reactivity with any of the other viruses tested, including bovine respiratory syncytial virus, bovine coronavirus, Akabane virus, bovine herpesvirus 1, bovine parainfluenza virus 3, bovine immunodeficiency virus, and bovine leukemia virus. The assay could detect the virus in serum samples with a titer as low as 10<sup>2</sup> TCID<sub>50</sub>/mL in two out of three trials for BVDV1 and all three trials for BVDV2, indicating that its sensitivity is equivalent to that of virus isolation. Our findings represent a significant advancement in BVDV detection and typing directly from bovine serum.DOI： 10.1007/s00705-024-06207-z
Other Link： https://link.springer.com/article/10.1007/s00705-024-06207-z/fulltext.html
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Complete genome sequence of the avian paramyxovirus serotype 9 strain duck/Miyazaki/128/2021. Reviewed International coauthorship International journal
Hirohisa Mekata, Mari Yamamoto, Yuto Matsui, Ahmad Massoud Niazi, Kentaro Yamada, Tamaki Okabayashi, Se-Yeoun Cha, Hyung-Kwan Jang
Microbiology resource announcements 13 ( 11 ) e0006024 2024.11
Authorship：Lead author, Corresponding author Language：English Publishing type：Research paper (scientific journal)
Here, we report the complete genome sequence of the avian paramyxovirus serotype 9 strain duck/Miyazaki/128/2021, which was determined using the Illumina MiSeq platform. The position of the hemagglutinin-neuraminidase stop codon differed from that of the only other available completely sequenced prototype strain, duck/New York/22/1977.
DOI： 10.1128/mra.00060-24
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Nine-year seroepidemiological study of severe fever with thrombocytopenia syndrome virus infection in feral horses in Cape Toi, Japan. Reviewed International journal
Hirohisa Mekata, Kentaro Yamada, Kazumi Umeki, Mari Yamamoto, Akihiro Ochi, Kunihiko Umekita, Ikuo Kobayashi, Takuya Hirai, Tamaki Okabayashi
BMC veterinary research 20 ( 1 ) 190 - 190 2024.5
Authorship：Lead author, Corresponding author Language：English Publishing type：Research paper (scientific journal)
Severe fever with thrombocytopenia syndrome (SFTS) is a fatal zoonosis caused by ticks in East Asia. As SFTS virus (SFTSV) is maintained between wildlife and ticks, seroepidemiological studies in wildlife are important to understand the behavior of SFTSV in the environment. Miyazaki Prefecture, Japan, is an SFTS-endemic area, and approximately 100 feral horses, called Misaki horses (Equus caballus), inhabit Cape Toi in Miyazaki Prefecture. While these animals are managed in a wild-like manner, their ages are ascertainable due to individual identification. In the present study, we conducted a seroepidemiological survey of SFTSV in Misaki horses between 2015 and 2023. This study aimed to understand SFTSV infection in horses and its transmission to wildlife. A total of 707 samples from 180 feral horses were used to determine the seroprevalence of SFTSV using enzyme-linked immunosorbent assay (ELISA). Neutralization testing was performed on 118 samples. In addition, SFTS viral RNA was detected in ticks from Cape Toi and feral horses. The overall seroprevalence between 2015 and 2023 was 78.5% (555/707). The lowest seroprevalence was 55% (44/80) in 2016 and the highest was 92% (76/83) in 2018. Seroprevalence was significantly affected by age, with 11% (8/71) in those less than one year of age and 96.7% (435/450) in those four years of age and older (p < 0.0001). The concordance between ELISA and neutralization test results was 88.9% (105/118). SFTS viral RNA was not detected in ticks (n = 516) or feral horses. This study demonstrated that horses can be infected with SFTSV and that age is a significant factor in seroprevalence in wildlife. This study provides insights into SFTSV infection not only in horses but also in wildlife in SFTS-endemic areas.
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MALDI-TOF MS analysis for detection of bovine coronavirus with tryptic peptides from viral proteins. Reviewed International journal
Katsuhiko Hayashi, Kenji Ohya, Tomoya Yoshinari, Shouhei Hirose, Souta Shimizu, Yuji Morita, Takahiro Ohnishi, Maiko Watanabe, Satoshi Taharaguchi, Hirohisa Mekata, Takahide Taniguchi, Yukiko Hara-Kudo
Journal of microorganism control 29 ( 4 ) 143 - 151 2024
Language：English Publishing type：Research paper (scientific journal)
Bovine coronavirus (BCoV）, a significant cattle pathogen causing enteric and respiratory diseases, is primarily detected using reverse transcription-polymerase chain reaction. Our objective was to develop a novel detection method for BCoV by matrix-assisted laser desorption/ionization‒time-of-flight mass spectrometry (MALDI-TOF MS）. Peptide mass fingerprint analysis revealed that nucleocapsid (N）, membrane (M）, and hemagglutinin-esterase (HE) were three main BCoV proteins. Their tryptic peptides were used as target molecules for BCoV detection. When the tryptic digest of 107.0 viral copies was analyzed by MALDI-TOF MS, five peptides with relatively strong peaks were detected. The detection limit was between 105.0 and 106.0 copies per test for BCoV alone. To detect BCoV in the swab eluate, ultrafiltration purification achieved a detection limit between 106.0 and 107.0 copies per test, sufficient to detect BCoV-infected calves. Our findings offer valuable insights for BCoV detection by MALDI-TOF MS.
DOI： 10.4265/jmc.29.4_143
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Generation of a Porcine Cell Line Stably Expressing Pig TMPRSS2 for Efficient Isolation of Swine Influenza Virus Reviewed International journal
Yuri L Tanaka, Maya Shofa, Erika P Butlertanaka, Ahmad Massoud Niazi, Takuya Hirai, Hirohisa Mekata, Akatsuki Saito
Pathogens 13 ( 1 ) 18 - 18 2023.12
Language：English Publishing type：Research paper (scientific journal) Publisher：MDPI AG
Pigs are important animals for meat production but can carry several zoonotic diseases, including the Japanese encephalitis virus, Nipah virus, and influenza viruses. Several Orthomyxoviridae and Coronavirinae respiratory viruses require cleavage of envelope proteins to acquire viral infectivity and consequently, need a host protease or the addition of exogenous trypsin for efficient propagation. Host TMPRSS2 is a key protease responsible for viral cleavage. Stable expression of human TMPRSS2 in African green monkey-derived Vero cells can enhance the porcine epidemic diarrhea virus. However, considering the narrow host tropism of viruses, a porcine cell line expressing pig TMPRSS2 could be optimal for replicating pig-derived viruses. Herein, we generated and evaluated a pig-derived PK-15 cell line stably expressing pig TMPRSS2. This cell line markedly (>1000-fold) and specifically enhanced the growth of influenza viruses. Furthermore, we demonstrated the usefulness of a PK-15 cell line lacking the Stat2 gene with a stable expression of pig TMPRSS2 for efficient virus isolation from clinical samples in the presence of type I interferons. Therefore, PK-15 cells expressing pig TMPRSS2 could be a valuable and promising tool for virus isolation, vaccine production, and virological studies of TMPRSS2-dependent viruses.
Books 【 display / non-display 】
MISC 【 display / non-display 】
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The Development of MALDI-ToF MS-based Analysis for the Direct Detection of SARS-CoV-2 in Nasopharyngeal Swabs
林克彦, 吉成知也, 廣瀬昌平, 大屋賢司, 大西貴弘, 渡辺麻衣子, 田原口智士, 目堅博久, 谷口隆秀, 前田卓哉, 折原悠太, 川村利江子, 新井沙倉, 斎藤嘉朗, 合田幸広, 工藤由起子
日本薬学会年会要旨集(Web) 143rd 2023
Language：Japanese Publishing type：Research paper, summary (national, other academic conference)
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牛伝染性リンパ腫の診断と早期発症予測法の開発
富永みその, 今内覚, 岡川朋弘, 神谷可菜, 齋藤麻矢, 安富一郎, 目堅博久, 前川直也, 村田史郎, 大橋和彦
北海道獣医師会雑誌 67 ( 8 ) 2023
Language：Japanese Publishing type：Research paper, summary (national, other academic conference)
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MALDI-ToF/MSによるSARS-CoV-2の新規検出法の開発
廣瀬昌平, 吉成知也, 林克彦, 大屋賢司, 大西貴弘, 渡辺麻衣子, 田原口智士, 目堅博久, 谷口隆秀, 新井沙倉, 合田幸広, 工藤由起子
日本獣医学会学術集会講演要旨集 165th (CD-ROM) 2022
Language：Japanese Publishing type：Research paper, summary (national, other academic conference)
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次世代シーケンス(NGS)を用いた豚糞便中のウイルス検索:新しいBastrovirus遺伝子の発見
長井誠, 岡林環樹, 松鵜彩, 藤本佳万, 目堅博久, 中尾亮, 浅井鉄夫, 中川敬介, 伊藤壽啓, 野中成晃, 小原恭子, 猪島康雄, 水谷哲也, 三澤尚明
日本獣医学会学術集会講演要旨集 163rd 2020
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Encouragement of bovine leukemia control measure based on bovine leukemia virus proviral load
Hirohisa MEKATA
The Journal of Farm Animal in Infectious Disease 7 ( 4 ) 163 - 168 2018.12
Language：Japanese Publishing type：Article, review, commentary, editorial, etc. (bulletin of university, research institution)
Presentations 【 display / non-display 】
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伴侶動物と野生動物のSFTS
目堅博久
第2回RCGLID&CADIC共同セミナー (大分大学) 2024.12.2 大分大学グローカル感染症研究センター（RCGLID）
Event date： 2024.12.2
Language：Japanese Presentation type：Public lecture, seminar, tutorial, course, or other speech
Venue：大分大学 Country：Japan
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牛伝染性リンパ腫対策について Invited
目堅博久
家畜防疫・衛生指導対策事業に関わる地域自衛防疫取組推進会議 (津市) 2024.9.26 三重県畜産協会
Event date： 2024.9.26
Language：Japanese Presentation type：Public lecture, seminar, tutorial, course, or other speech
Venue：津市 Country：Japan
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2021-23年度に宮崎⼤学で実施した伴侶動物の SFTS 検査
目堅博久, 金子泰之, 平井卓哉, 梅木一美, 梅北邦彦, 山田健太郎, 岡林環樹
第6回SFTS研究会 (札幌市) 2024.9.14 SFTS研究会
Event date： 2024.9.14 - 2024.9.15
Language：Japanese Presentation type：Oral presentation (general)
Venue：札幌市 Country：Japan
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Urine from cats with severe fever with thrombocytopenia syndrome could be an infectious source International conference
Hirohisa Mekata, Yasuyuki Kaneko, Kentaro Yamada, Tamaki Okabayashi, Akatsuki Saito
The 4th Joint Meeting of Veterinary Science in East Asia (Obihiro) 2024.9.8 The Japanese Society of Veterinary Science
Event date： 2024.9.8 - 2024.9.9
Language：English Presentation type：Oral presentation (general)
Venue：Obihiro Country：Japan
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プロウイルス量を使った繁殖農場でのBLV対策と課題 Invited
目堅博久
第64回獣医疫学会学術集会(シンポジウム) (オンライン) 2024.8.31 獣医疫学会
Event date： 2024.8.31
Language：Japanese Presentation type：Symposium, workshop panel (nominated)
Venue：オンライン Country：Japan
Awards 【 display / non-display 】
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日本獣医学会獣医学奨励賞
2013.3 日本獣医学会
目堅 博久
Award type：Award from Japanese society, conference, symposium, etc. Country：Japan
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第１５５回日本獣医学会大会長賞
2013.3 日本獣医学会
目堅 博久
Award type：Award from Japanese society, conference, symposium, etc. Country：Japan
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日本産業動物獣医学会九州地区学会長賞
2014.10 日本産業動物獣医学会
目堅 博久
Award type：Award from Japanese society, conference, symposium, etc. Country：Japan
Grant-in-Aid for Scientific Research 【 display / non-display 】
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動物フォーミーウイルスベクターの基盤構築と牛伝染性リンパ腫ワクチンの開発
Grant number：23K23765 2024.04 - 2026.03
独立行政法人日本学術振興会 科学研究費基金 基盤研究(B)
Authorship：Principal investigator
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ベトナムにおけるアフリカ豚熱の実践的な高感度診断法の開発と環境動態の解明
Grant number：22KK0097 2022.04 - 2027.03
独立行政法人日本学術振興会 科学研究費基金 国際共同研究加速基金(国際共同研究強化(B))
Authorship：Coinvestigator(s)
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Construction of animal foamy virus vector and development of enzootic bovine leukosis vaccine
Grant number：22H02500 2022.04 - 2026.03
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
Authorship：Principal investigator
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Studies on anti-tumor potential of symbiotic retroviruses and their therapeutic applications
Grant number：20H03150 2020.04 - 2024.03
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)
Authorship：Coinvestigator(s)
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Pathological analysis of nasopharynx and lung caused by respiratory disease virus in pigs
Grant number：18K05974 2018.04 - 2021.03
Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
Hirai Takuya
Authorship：Coinvestigator(s)
Swine influenza virus (A/H1N1pdm09) were detected in the piglets of the local pig farm in Miyazaki prefecture. Subclinical infection was considered because these piglets did not show respiratory sign. CK-18 immunohistochemistry demonstrated M cells in the epithelial cells covering nasal-associated lymphoid tissue (NALT). In addition, lectin histochemistry revealed that the number of α2,6-linked sialic acid is more than α2,3-linked sialic acid in the epithelial cells of NALT. Several epithelial cells show double positive for both CK-18 immunohistochemistry and α2,6-linked sialic acid lectin histochemistry. Influenza positive macrophages were also detected in the NALT. These results suggest that M cells express α2,6-linked sialic acid, and influenza virus transported by M cells may be engulfed by macrophages or dendritic cells.