顔写真

サノ　ダイスケ
佐野　大輔
Daisuke Sano
所属
大学院工学研究科　土木工学専攻　水環境学講座（環境水質工学分野）
職名
教授
学位

  • 博士（工学）（東北大学）

  • 修士（工学）（東北大学）

経歴 9

  • 2021年4月 ～ 継続中
    東北大学　大学院工学研究科 土木工学専攻　教授

  • 2015年4月 ～ 継続中
    北九州市立大学　非常勤講師

  • 2018年4月 ～ 2021年3月
    東北大学　大学院環境科学研究科 先端環境創成学専攻　准教授

  • 2017年4月 ～ 2018年3月
    東北大学　大学院工学研究科 土木工学専攻　准教授

  • 2009年4月 ～ 2017年3月
    北海道大学　大学院工学研究院　准教授

  • 2015年5月 ～ 2015年6月
    バルセロナ大学　生物学部微生物学科　客員准教授

  • 2007年4月 ～ 2009年3月
    バルセロナ大学　生物学部微生物学科　日本学術振興会海外特別研究員

  • 2003年4月 ～ 2007年3月
    東北大学　大学院工学研究科　日本学術振興会特別研究員 (PD)

  • 2002年4月 ～ 2003年3月
    東北大学　大学院工学研究科　日本学術振興会特別研究員（DC2）

︎全件表示 ︎最初の5件までを表示

学歴 6

  • 東北大学　工学研究科　土木工学専攻

    ～ 2003年

  • 東北大学

    ～ 2003年

  • 東北大学　工学研究科　土木工学専攻

    ～ 2000年

  • 東北大学

    ～ 2000年

  • 東北大学　工学部　土木工学科

    ～ 1998年

  • 東北大学

    ～ 1998年

︎全件表示 ︎最初の5件までを表示

委員歴 25

  • 日本水環境学会　理事

    2021年7月 ～ 継続中

  • Health-Related Water Microbiology study group (IWA)　Secretary

    2017年5月 ～ 継続中

  • 平成２７年度総合地球環境学研究所　共同研究員

    2015年10月 ～ 継続中

  • 日本水環境学会・水中健康関連微生物研究委員会　幹事

    2014年4月 ～ 継続中

  • Journal of Water and Health　Editor

    2013年10月 ～ 継続中

  • 日本水環境学会　運営幹事

    2013年8月 ～ 2021年6月

  • Water and Environment Technology Conference　Secretary

    2013年11月 ～ 2017年8月

  • Health-Related Water Microbiology study group (IWA)　Steering committee member

    2013年9月 ～ 2017年5月

  • 土木学会・環境工学委員会　次世代下水道小委員会委員

    2015年6月 ～ 2017年3月

  • 土木学会・環境工学委員会　副幹事長

    2015年4月 ～ 2017年3月

  • 北海道大学大学院工学研究院　国際交流室

    2014年4月 ～ 2017年3月

  • 北海道大学大学院工学研究院　ヒトを対象とする研究倫理審査委員会

    2014年4月 ～ 2017年3月

  • 日本水環境学会北海道支部　幹事

    2013年4月 ～ 2017年3月

  • 土木学会　環境工学委員会

    2011年4月 ～ 2017年3月

  • 46. International Symposium for Food and Environmental Virology 2016　Secretary of Organizing Committee

    2015年2月 ～ 2016年10月

  • Sapporo Pioneer Toastmaster Club　Vice President Membership

    2015年7月 ～ 2016年6月

  • 経済産業省北海道経済産業局・ミルキングパーラー排水の低コスト排水処理施設モデル構築のための検討ワーキング会議　委員長

    2015年9月 ～ 2016年3月

  • 札幌市　環境影響評価審議会委員

    2014年9月 ～ 2015年8月

  • Sapporo Pioneer Toastmaster Club　President

    2014年7月 ～ 2015年6月

  • 北海道大学工学部・日韓共同理工系学部留学生等専門部会　座長

    2014年5月 ～ 2015年3月

  • 土木学会・環境工学委員会　今後の水環境保全に貢献する下水道システムの技術的課題と管理手法についての調査研究に関する小委員会

    2012年6月 ～ 2015年3月

  • 経済産業省北海道経済産業局・北海道根釧地域における環境対策支援のための検討ワーキング会議　委員長

    2014年8月 ～ 2015年2月

  • Sapporo Pioneer Toastmaster Club　Vise President Education

    2013年7月 ～ 2014年6月

  • 土木学会論文集編集委員会　G分冊編集小委員会

    2012年6月 ～ 2014年5月

  • 土木学会・環境工学委員会　下水道関連震災調査小委員会

    2012年2月 ～ 2013年3月

︎全件表示 ︎最初の5件までを表示

所属学協会 8

  • ウイルス性下痢症研究会

  • 日本ウイルス学会

  • 臨床ウイルス学会

  • American Chemical Society

  • International Water Association

  • American Society for Microbiology

  • 日本水環境学会

  • 土木学会

︎全件表示 ︎最初の5件までを表示

研究キーワード 7

  • ノロウイルス

  • 腸内細菌

  • 水環境中動態

  • 消毒

  • ウイルス吸着タンパク質

  • 感染症

  • Civil and Environmental Engineering

研究分野 4

  • ライフサイエンス / 衛生学、公衆衛生学分野：実験系を含まない /

  • ライフサイエンス / 衛生学、公衆衛生学分野：実験系を含む /

  • ライフサイエンス / 医療管理学、医療系社会学 /

  • 社会基盤（土木・建築・防災） / 土木環境システム /

受賞 21

  1. 論文賞

    2018年6月　日本水環境学会　Bacterial histo-blood group antigens contributing to genotype-dependent removal of human noroviruses with a microfiltration membrane

  2. 総長研究賞（奨励賞）

    2016年2月　北海道大学

  3. 若手奨励賞

    2014年6月　日本臨床ウイルス学会

  4. 第50回環境工学研究フォーラム 優秀ポスター発表賞

    2013年11月　土木学会・環境工学委員会

  5. 第49回環境工学研究フォーラム 優秀ポスター発表賞

    2012年11月　土木学会・環境工学委員会

  6. メタウォーター賞（年間優秀論文賞）

    2011年9月　日本水環境学会

  7. 文部科学大臣表彰 若手科学者賞

    2008年

  8. The Commendation for Science and Technology by the Minister of Education, Culture, Sports, Science and Technology, The Young Scientists' Prize

    2008年

  9. Presentation Award in the 7th meeting of Water Microbiology Specialist Group in Spanish Society of Microbiology

    2008年

  10. 土木学会 環境工学研究フォーラム論文奨励賞

    2007年

  11. 土木学会 論文奨励賞

    2007年

  12. Encourage paper award from Japan Society for Civil Engineers

    2007年

  13. Encourage paper award from Committee of Environmental Engineering, Japan Society for Civil Engineers

    2007年

  14. 土木学会東北支部 技術開発賞

    2006年

  15. Technological development award from northeast area chapter of Japan Society for Civil Engineers

    2006年

  16. 土木学会 環境工学研究フォーラム論文賞

    2005年

  17. Excellent paper award from Committee of Environmental Engineering, Japan Society for Civil Engineers

    2005年

  18. 土木学会 優秀講演者表彰

    2004年9月

  19. Best presentation award from Japan Society for Civil Engineers

    2004年

  20. Grant for the research presentation in abroad by the Fundation for promotion of sewage system

    2002年

  21. Sato Atsuhisa Fund for young researcher

    2000年

︎全件表示 ︎最初の5件までを表示

論文 133

  1. Decay rate estimation of respiratory viruses in aerosols and on surfaces under different environmental conditions

    Sewwandi Bandara, Wakana Oishi, Syun-suke Kadoya, Daisuke Sano

    International Journal of Hygiene and Environmental Health　251　114187-114187　2023年6月

    出版者・発行元：Elsevier BV

    DOI： 10.1016/j.ijheh.2023.114187 　

    ISSN：1438-4639

  2. Temporal variations of human and animal Rotavirus A genotypes in surface water used for drinking water production

    Takayuki Miura, Syun-suke Kadoya, Hiroyuki Takino, Daisuke Sano, Michihiro Akiba

    Frontiers in Microbiology　13　2022年8月9日

    出版者・発行元：Frontiers Media SA

    DOI： 10.3389/fmicb.2022.912147 　

    eISSN：1664-302X

    詳細を見る 詳細を閉じる

    Rotavirus is a major cause of gastroenteritis among infants and children. In this study, nested PCR assays were developed to amplify partial regions of the VP7, VP4, and VP6 genes of Rotavirus A (RVA) for amplicon-based Illumina MiSeq sequencing to investigate RVA genotypes in environmental water samples. Eight sets of inner primers were first designed and screened for use in the nested PCR assays, and four sets of them could produce amplicons. Six sets of outer primers were then designed and combined with the four sets of inner primers that worked. The assays were evaluated for sensitivity using raw water samples collected from one drinking water treatment plant between April 2019 and March 2020 (Sample Set 1; N = 12) and seven DWTPs between 2018 and 2020 (Sample Set 2; N = 18). In total, 43 amplicons from Set 1 were sequenced and diverse sequences from human, porcine, bovine, equine, and feline RVA were observed. Human G8, G3, and G2 genotypes were obtained, with G8 predominant (relative abundance, 36–87%) in samples taken during the rotavirus epidemic season between April and June. Porcine G5, G11, and G4, and bovine G10 and G6 genotypes were also detected. VP4 sequence analysis revealed that the human P[8] genotype was present throughout the year, whereas P[4] and P[9] were present only in the epidemic season. The vaccine strains P[5] and P[8] (RotaTeq<sup>®</sup>) were also detected. Our approach enables the identification of prevalent human and animal RVA genotypes and their host species that potentially caused fecal contamination in water sources.

  3. Unveiling the characterization and development of prokaryotic community during the start-up and long-term operation of a pilot-scale anaerobic membrane bioreactor for the treatment of real municipal wastewater 査読有り

    Zhe Kong, Lu Li, Jiang Wu, Chao Rong, Tianjie Wang, Rong Chen, Daisuke Sano, Yu-You Li

    813　152643-152643　2022年3月

    出版者・発行元：None

    DOI： 10.1016/j.scitotenv.2021.152643 　

    ISSN：0048-9697

    eISSN：1879-1026

  4. Comparison of five polyethylene glycol precipitation procedures for the RT-qPCR based recovery of murine hepatitis virus, bacteriophage phi6, and pepper mild mottle virus as a surrogate for SARS-CoV-2 from wastewater 国際誌 査読有り

    Shotaro Torii, Wakana Oishi, Yifan Zhu, Ocean Thakali, Bikash Malla, Zaizhi Yu, Bo Zhao, Chisato Arakawa, Masaaki Kitajima, Akihiko Hata, Masaru Ihara, Shigeru Kyuwa, Daisuke Sano, Eiji Haramoto, Hiroyuki Katayama

    807　(Pt 2)　150722-150722　2022年2月

    出版者・発行元：None

    DOI： 10.1016/j.scitotenv.2021.150722 　

    ISSN：0048-9697

  5. The Intrapopulation Genetic Diversity of RNA Virus May Influence the Sensitivity of Chlorine Disinfection. 国際誌

    Syun-Suke Kadoya, Syun-Ichi Urayama, Takuro Nunoura, Miho Hirai, Yoshihiro Takaki, Masaaki Kitajima, Toyoko Nakagomi, Osamu Nakagomi, Satoshi Okabe, Osamu Nishimura, Daisuke Sano

    Frontiers in microbiology　13　839513-839513　2022年

    DOI： 10.3389/fmicb.2022.839513 　

    詳細を見る 詳細を閉じる

    RNA virus populations are not clonal; rather, they comprise a mutant swarm in which sequences are slightly different from the master sequence. Genetic diversity within a population (intrapopulation genetic diversity) is critical for RNA viruses to survive under environmental stresses. Disinfection has become an important practice in the control of pathogenic viruses; however, the impact of intrapopulation genetic diversity on the sensitivity of disinfection, defined as -log10 (postdisinfected infectious titer/predisinfected titer), has not been elucidated. In this study, we serially passaged populations of rhesus rotavirus. We demonstrated that populations with reduced chlorine sensitivity emerged at random and independently of chlorine exposure. Sequencing analysis revealed that compared with sensitive populations, less-sensitive ones had higher non-synonymous genetic diversity of the outer capsid protein gene, suggesting that changes in the amino acid sequences of the outer capsid protein were the main factors influencing chlorine sensitivity. No common mutations were found among less-sensitive populations, indicating that rather than specific mutations, the diversity of the outer capsid protein itself was associated with the disinfection sensitivity and that the disinfection sensitivity changed stochastically. Simulation results suggest that the disinfection sensitivity of a genetically diverse population is destabilized if cooperative viral clusters including multiple sequences are formed. These results advocate that any prevention measures leading to low intrapopulation genetic diversity are important to prevent the spread and evolution of pathogenic RNA viruses in society.

  6. Experimental Adaptation of Murine Norovirus to Calcium Hydroxide. 国際誌

    Wakana Oishi, Mikiko Sato, Kengo Kubota, Ryoka Ishiyama, Reiko Takai-Todaka, Kei Haga, Kazuhiko Katayama, Daisuke Sano

    Frontiers in microbiology　13　848439-848439　2022年

    DOI： 10.3389/fmicb.2022.848439 　

    詳細を見る 詳細を閉じる

    Slaked lime (calcium hydroxide) is a commonly used disinfectant for fecal sludge. Although viruses are inactivated by lime treatment, whether RNA viruses adapt to lime treatment has not yet been determined. Here, we show that murine norovirus developed higher tolerance during serial passages with lime treatment. We compared synonymous and non-synonymous nucleotide diversities of the three open reading frames of viral genome and revealed that virus populations were subjected to enhanced purifying selection over the course of serial passages with lime treatment. Virus adaptation to lime treatment was coincident with amino acid substitution of lysine to arginine at position 345 (K345R) on the major capsid protein VP1, which accounted for more than 90% of the population. The infectious clones with the K345R produced using a plasmid-based reverse genetics system exhibited greater tolerance in a lime solution, which indicated that the specific amino acid substitution was solely involved in the viral tolerance in lime treatment.

  7. Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance 査読有り

    Warish Ahmed, Stuart L. Simpson, Paul M. Bertsch, Kyle Bibby, Aaron Bivins, Linda L. Blackall, Sílvia Bofill-Mas, Albert Bosch, João Brandão, Phil M. Choi, Mark Ciesielski, Erica Donner, Nishita D'Souza, Andreas H. Farnleitner, Daniel Gerrity, Raul Gonzalez, John F. Griffith, Pradip Gyawali, Charles N. Haas, Kerry A. Hamilton, Hapuarachchige Chanditha Hapuarachchi, Valerie J. Harwood, Rehnuma Haque, Greg Jackson, Stuart J. Khan, Wesaal Khan, Masaaki Kitajima, Asja Korajkic, Giuseppina La Rosa, Blythe A. Layton, Erin Lipp, Sandra L. McLellan, Brian McMinn, Gertjan Medema, Suzanne Metcalfe, Wim G. Meijer, Jochen F. Mueller, Heather Murphy, Coleen C. Naughton, Rachel T. Noble, Sudhi Payyappat, Susan Petterson, Tarja Pitkänen, Veronica B. Rajal, Brandon Reyneke, Fernando A. Roman, Joan B. Rose, Marta Rusiñol, Michael J. Sadowsky, Laura Sala-Comorera, Yin Xiang Setoh, Samendra P. Sherchan, Kwanrawee Sirikanchana, Wendy Smith, Joshua A. Steele, Rosalie Sabburg, Erin M. Symonds, Phong Thai, Kevin V. Thomas, Josh Tynan, Simon Toze, Janelle Thompson, Andy S. Whiteley, Judith Chui Ching Wong, Daisuke Sano, Stefan Wuertz, Irene Xagoraraki, Qian Zhang, Amity G. Zimmer-Faust, Orin C. Shanks

    805　149877-149877　2022年1月

    出版者・発行元：None

    DOI： 10.1016/j.scitotenv.2021.149877 　

    ISSN：0048-9697

  8. Disinfection efficacy and mechanism of olanexidine gluconate against norovirus 査読有り

    Kaoru Imai, Makoto Tanaka, Seiji Miyoshi, Ren Murakami, Akifumi Hagi, Sachi Yamagawa, Daisuke Sano

    2021年12月

    出版者・発行元：None

    DOI： 10.1016/j.ajic.2021.11.020 　

    ISSN：0196-6553

  9. Predictive Environmental Microbiology for Safe Use of Sanitation Products in Agriculture: Challenges and Perspectives 査読有り

    Wakana Oishi, Björn Vinnerås, Joan B. Rose, Daisuke Sano

    8　(11)　924-931　2021年11月9日

    出版者・発行元：None

    DOI： 10.1021/acs.estlett.1c00537 　

    ISSN：2328-8930

    eISSN：2328-8930

  10. The water temperature changes the effect of pH on copper toxicity to the green microalgae Raphidocelis subcapitata 査読有り

    Gissela Pascual, Daisuke Sano, Takashi Sakamaki, Michihiro Akiba, Osamu Nishimura

    133110-133110　2021年11月

    出版者・発行元：None

    DOI： 10.1016/j.chemosphere.2021.133110 　

    ISSN：0045-6535

  11. ウイルス感染症流行制御のための下水疫学調査のエッセンス 査読有り

    大石若菜, Zhu Yifan, 丸尾知佳子, 斉藤繭子, 北島正章, 佐野大輔

    水環境学会誌　44　(5)　125-133　2021年9月

  12. Early warning of COVID-19 in Tokyo via wastewater-based epidemiology: How feasible it really is? 査読有り

    Yifan Zhu, Wakana Oishi, Mayuko Saito, Masaaki Kitajima, Daisuke Sano

    Journal of Water and Environment Technology　19　(3)　170-183　2021年6月

    出版者・発行元：None

    DOI： 10.2965/jwet.21-024 　

    eISSN：1348-2165

  13. Olanexidine gluconate formulations as environmental disinfectants for enveloped viruses infection control 査読有り

    Kaoru Imai, Akifumi Hagi, Sachi Yamagawa, Daisuke Sano

    Journal of Hospital Infection　112　37-41　2021年6月

  14. Virus disinfection and population genetics: Toward the control of waterborne virus diseases by water engineering 査読有り

    Syun-suke Kadoya, Hiroyuki Katayama, Daisuke Sano

    Current Pollution Reports　7　407-416　2021年5月

  15. Predictive water virology using regularized regression analyses for projecting virus inactivation efficiency in ozone disinfection 査読有り

    Syun-suke Kadoya, Osamu Nishimura, Hiroyuki Kato, Daisuke Sano

    Water Research X　11　100093　2021年5月

  16. Early warning of COVID-19 via wastewater-based epidemiology: potential and bottlenecks 国際誌 査読有り

    Yifan Zhu, Wakana Oishi, Chikako Maruo, Mayuko Saito, Rong Chen, Masaaki Kitajima, Daisuke Sano

    Science of the Total Environment　767　145124-145124　2021年5月

    DOI： 10.1016/j.scitotenv.2021.145124 　

    詳細を見る 詳細を閉じる

    An effective early warning tool is of great administrative and social significance to the containment and control of an epidemic. Facing the unprecedented global public health crisis caused by COVID-19, wastewater-based epidemiology (WBE) has been given high expectations as a promising surveillance complement to clinical testing which had been plagued by limited capacity and turnaround time. In particular, recent studies have highlighted the role WBE may play in being a part of the early warning system. In this study, we briefly discussed the basics of the concept, the benefits and critical points of such an application, the challenges faced by the scientific community, the progress made so far, and what awaits to be addressed by future studies to make the concept work. We identified that the shedding dynamics of infected individuals, especially in the form of a mathematical shedding model, and the back-calculation of the number of active shedders from observed viral load are the major bottlenecks of WBE application in the COVID-19 pandemic that deserve more attention, and the sampling strategy (location, timing, and interval) needs to be optimized to fit the purpose and scope of the WBE project.

  17. Hierarchical Bayesian modeling for predictive environmental microbiology toward a safe use of human excreta: Systematic review and meta-analysis 査読有り

    Wakana Oishi, Syun-suke Kadoya, Osamu Nishimura, Joan B. Rose, Daisuke Sano

    Journal of Environmental Management　284　112088　2021年4月

  18. Biochar and GAC intensify anaerobic phenol degradation via distinctive adsorption and conductive properties 査読有り

    Qian Li, Xin Gao, Yaqian Liu, Gaojun Wang, Yu-You Li, Daisuke Sano, Xiaochang Wang, Rong Chen

    Journal of Hazardous Materials　124183-124183　2021年3月

    出版者・発行元：Elsevier BV

    DOI： 10.1016/j.jhazmat.2020.124183 　

    ISSN：0304-3894

  19. Human norovirus disease burden of consuming Crassostrea gigas oysters: A case-study from Japan 査読有り

    Yo Ueki, Mohan Amarasiri, Sayaka Kamio, Akie Sakagami, Hiroshi Ito, Sital Uprety, Arief Nurul Umam, Takayuki Miura, Thanh H. Nguyen, Daisuke Sano

    Food Control　121　107556-107556　2021年3月

    出版者・発行元：Elsevier BV

    DOI： 10.1016/j.foodcont.2020.107556 　

    ISSN：0956-7135

  20. Corrigendum to “Effects of chemical interaction of nutrients and EDTA on metals toxicity to Pseudokirckneriella subcapitata” [Ecotoxicol. Environ. Saf. 203 (2020) 110966] (Ecotoxicology and Environmental Safety (2020) 203, (S0147651320308058), (10.1016/j.ecoenv.2020.110966))

    Gissela Pascual, Daisuke Sano, Takashi Sakamaki, Osamu Nishimura

    Ecotoxicology and Environmental Safety　208　2021年1月15日

    DOI： 10.1016/j.ecoenv.2020.111465 　

    ISSN：0147-6513

    eISSN：1090-2414

    詳細を見る 詳細を閉じる

    The authors regret a misspelling in the title of our manuscript. The name of the selected microalgae species for the experiments is “Pseudokirchneriella subcapitata” instead of “Pseudokirckneriella subcapitata”. The authors would like to apologise for any inconvenience caused.

  21. Viral Interference as a Factor of False-Negative in the Infectious Adenovirus Detection Using Integrated Cell Culture-PCR with a BGM Cell Line 査読有り

    Daisuke Sano, Ryosuke Watanabe, Wakana Oishi, Mohan Amarasiri, Masaaki Kitajima, Satoshi Okabe

    Food and Environmental Virology　13　84-91　2021年1月

  22. Virus removal by membrane bioreactors: A review of mechanism investigation and modeling efforts 査読有り

    Yifan Zhu, Rong Chen, Yu-You Li, Daisuke Sano

    Water Research　188　116522-116522　2021年1月

    出版者・発行元：Elsevier BV

    DOI： 10.1016/j.watres.2020.116522 　

    ISSN：0043-1354

  23. スパース推定法と階層ベイズ推定法による環境水中ウイルス自然死滅モデルの構築 査読有り

    大石若菜, 加藤郁生, 西村修, 佐野大輔

    土木学会論文集　76　(6)　III_449-III_460　2020年12月

  24. Updated research agenda for water, sanitation and antimicrobial resistance 査読有り

    Daisuke Sano, Astrid Louse Wester, Heike Schmitt, Mohan Amarasiri, Amy Kirby, Kate Medlicott, Ana Maria de, Roda Husman

    Journal of Water and Health　18　(6)　858-868　2020年12月

  25. The Effect of GD1a Ganglioside-Expressing Bacterial Strains on Murine Norovirus Infectivity 国際誌 査読有り

    Yifan Zhu, Hiroki Kawai, Satoshi Hashiba, Mohan Amarasiri, Masaaki Kitajima, Satoshi Okabe, Daisuke Sano

    Molecules　25　(18)　4084-4084　2020年9月7日

    出版者・発行元：MDPI AG

    DOI： 10.3390/molecules25184084 　

    eISSN：1420-3049

    詳細を見る 詳細を閉じる

    In this study, we investigated the impact of GD1a-expressing bacterial strains on the infectivity of murine norovirus (MNV). Eligible bacterial strains were screened from a sewage sample using flow cytometry, and their genetic sequences of 16S rRNA were determined. The enzyme-linked immunosorbent assay (ELISA) was employed to analyze the binding between bacteria and MNV particles, and the plaque assay was used to assess the effects of GD1a-positive and negative strains on MNV infectivity. The result from ELISA shows that MNV particles are able to bind to both GD1a-positive and negative bacterial strains, but the binding to the GD1a-positive strain is more significant. The infectivity assay result further shows that the MNV infectious titer declined with an increasing concentration of GD1a-positive bacteria. The addition of anti-GD1a antibody in the infectivity assay led to the recovery of the MNV infectious titer, further confirming that the binding between MNV particles and bacterial GD1a ganglioside compromises MNV infectivity. Our findings highlight the role indigenous bacteria may play in the lifecycle of waterborne enteric viruses as well as the potential of exploiting them for virus transmission intervention and water safety improvement.

  26. Inactivation kinetics modeling of Escherichia coli in concentrated urine for implementing predictive environmental microbiology in sanitation safety planning. 国際誌 査読有り

    Wakana Oishi, Ikuo Kato, Nowaki Hijikata, Ken Ushijima, Ryusei Ito, Naoyuki Funamizu, Osamu Nishimura, Daisuke Sano

    Journal of Environmental Management　268　110672-110672　2020年8月15日

    DOI： 10.1016/j.jenvman.2020.110672 　

  27. Identification of novel norovirus polymerase genotypes from pediatric fecal samples collected between the year 1997 and 2000 in Japan. 国際誌 査読有り

    Mohan Amarasiri, Etsuko Utagawa, Daisuke Sano, Kazuhiko Katayama

    Infection, Genetics and Evolution　82　104313-104313　2020年8月

    DOI： 10.1016/j.meegid.2020.104313 　

  28. Effects of chemical interaction of nutrients and EDTA on metals toxicity to Pseudokirckneriella subcapitata. 国際誌 査読有り

    Gissela Pascual, Daisuke Sano, Takashi Sakamaki, Osamu Nishimura

    Ecotoxicology and Environmental Safety　203　110966-110966　2020年7月14日

    出版者・発行元：None

    DOI： 10.1016/j.ecoenv.2020.110966 　

    ISSN：0147-6513

  29. Virucidal efficacy of olanexidine gluconate as a hand antiseptic against human norovirus 国際誌 査読有り

    Kaoru Imai, Akifumi Hagi, Yasuhide Inoue, Mohan Amarasiri, Daisuke Sano

    Food and Environmental Virology　12　(2)　180-190　2020年6月

    出版者・発行元：None

    DOI： 10.1007/s12560-020-09422-4 　

    ISSN：1867-0334

    eISSN：1867-0342

  30. Improvement of Electrochemical Conditions for Detecting Redox Reaction of K<sub>3</sub>[Fe(CN)<sub>6</sub>] toward the Application in Norovirus Aptasensor 査読有り

    Seiya HIRANO, Junki SAITO, Tomoki YUKAWA, Daisuke SANO, Akihiro OKAMOTO, Satoshi OKABE, Masaaki KITAJIMA

    Electrochemistry　88　(3)　205-209　2020年5月5日

    出版者・発行元：The Electrochemical Society of Japan

    DOI： 10.5796/electrochemistry.20-00017 　

    ISSN：1344-3542

    eISSN：2186-2451

  31. Bottleneck size-eependent changes in the genetic diversity and specific growth rate of a rotavirus A strain 国際誌 査読有り

    Syun-Suke Kadoya, Syun-Ichi Urayama, Takuro Nunoura, Miho Hirai, Yoshihiro Takaki, Masaaki Kitajima, Toyoko Nakagomi, Osamu Nakagomi, Satoshi Okabe, Osamu Nishimura, Daisuke Sano

    Journal of Virology　94　(10)　e02083-19　2020年5月4日

    DOI： 10.1128/JVI.02083-19 　

  32. Assessment of microbial risks by characterization of Escherichia coli presence to analyze the public health risks from poor water quality in Nepal 国際誌 査読有り

    Sital Uprety, Bipin Dangol, Pramina Nakarmi, Isha Dhakal, Samendra P Sherchan, Joanna L Shisler, Antarpreet Jutla, Mohan Amarasiri, Daisuke Sano, Thanh H Nguyen

    International Journal of Hygiene and Environmental Health　226　113484-113484　2020年5月

    DOI： 10.1016/j.ijheh.2020.113484 　

  33. Required chlorination doses to fulfill the credit value for disinfection of enteric viruses in water: A critical review 国際誌 査読有り

    Andri Taruna Rachmadi, Masaaki Kitajima, Tsuyoshi Kato, Hiroyuki Kato, Satoshi Okabe, Daisuke Sano

    Environmental Science & Technology　54　(4)　2068-2077　2020年2月18日

    DOI： 10.1021/acs.est.9b01685 　

  34. Regularized regression analysis for the prediction of virus inactivation efficiency by chloramine disinfection 査読有り

    Syun-suke Kadoya, Osamu Nishimura, Hiroyuki Kato, Daisuke Sano

    Environmental Science: Water Research & Technology　6　(12)　3341-3350　2020年

    出版者・発行元：Royal Society of Chemistry (RSC)

    DOI： 10.1039/d0ew00539h 　

    ISSN：2053-1400

    eISSN：2053-1419

    詳細を見る 詳細を閉じる

    <p>The sparse modeling methods using water quality information as explanatory variables enable us to appropriately predict virus inactivation efficiency in wastewater treatment plants.</p>

  35. Understanding human health risks caused by antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARG) in water environments: Current knowledge and questions to be answered 査読有り

    Mohan Amarasiri, Daisuke Sano, Satoru Suzuki

    Critical Reviews in Environmental Science and Technology　50　(19)　2016-2059　2020年

    出版者・発行元：None

    DOI： 10.1080/10643389.2019.1692611 　

  36. Revisiting the effects of powdered activated carbon on membrane fouling mitigation in an anaerobic membrane bioreactor by evaluating long-term impacts on the surface layer 国際誌 査読有り

    Zhen Lei, Shuming Yang, Xiang Li, Wen Wen, Xingyuan Huang, Yuan Yang, Xiaochang Wang, Yu-You Li, Daisuke Sano, Rong Chen

    Water Research　167　115137-115137　2019年12月15日

    DOI： 10.1016/j.watres.2019.115137 　

  37. 各戸導入型小型水供給設備の利用における水安全計画的アプローチによる健康リスク管理 査読有り

    門屋俊祐, 牛島健, 伊藤竜生, 長谷川祥樹, 三浦尚之, 秋葉道宏, 西村修, 佐野大輔

    土木学会論文集　75　(7)　III_403-III_412　2019年12月

  38. Predictive water virology: Hierarchical Bayesian modeling for estimating virus inactivation curve 査読有り

    Syun-suke Kadoya, Osamu Nishimura, Hiroyuki Kato, Daisuke Sano

    Water　11　(10)　2187　2019年10月

  39. Fecal Source Tracking in A Wastewater Treatment and Reclamation System Using Multiple Waterborne Gastroenteritis Viruses. 国際誌 査読有り

    Zheng Ji, Xiaochang C Wang, Limei Xu, Chongmiao Zhang, Cheng Rong, Andri Taruna Rachmadi, Mohan Amarasiri, Satoshi Okabe, Naoyuki Funamizu, Daisuke Sano

    Pathogens (Basel, Switzerland)　8　(4)　170-170　2019年9月30日

    出版者・発行元：{MDPI} {AG}

    DOI： 10.3390/pathogens8040170 　

    詳細を見る 詳細を閉じる

    Gastroenteritis viruses in wastewater reclamation systems can pose a major threat to public health. In this study, multiple gastroenteritis viruses were detected from wastewater to estimate the viral contamination sources in a wastewater treatment and reclamation system installed in a suburb of Xi'an city, China. Reverse transcription plus nested or semi-nested PCR, followed by sequencing and phylogenetic analysis, were used for detection and genotyping of noroviruses and rotaviruses. As a result, 91.7% (22/24) of raw sewage samples, 70.8% (17/24) of the wastewater samples treated by anaerobic/anoxic/oxic (A2O) process and 62.5% (15/24) of lake water samples were positive for at least one of target gastroenteritis viruses while all samples collected from membrane bioreactor effluent after free chlorine disinfection were negative. Sequence analyses of the PCR products revealed that epidemiologically minor strains of norovirus GI (GI/14) and GII (GII/13) were frequently detected in the system. Considering virus concentration in the disinfected MBR effluent which is used as the source of lake water is below the detection limit, these results indicate that artificial lake may be contaminated from sources other than the wastewater reclamation system, which may include aerosols, and there is a possible norovirus infection risk by exposure through reclaimed water usage and by onshore winds transporting aerosols containing norovirus.

  40. Sign-constrained linear regression for prediction of microbe concentration based on water quality datasets 国際誌 査読有り

    Tsuyoshi Kato, Ayano Kobayashi, Wakana Oishi, Syun-suke Kadoya, Satoshi Okabe, Naoya Ohta, Mohan Amarasiri, Daisuke Sano

    Journal of Water and Health　17　(3)　404-415　2019年6月

    DOI： 10.2166/wh.2019.001 　

  41. Specific interactions between human norovirus and environmental matrices: Effects on the virus ecology 査読有り

    Mohan Amarasiri, Daisuke Sano

    Viruses　11　(3)　224　2019年3月

  42. Specific interactions of rotavirus HAL1166 with Enterobacter cloacae SENG-6 and their contribution on rotavirus HAL1166 removal 査読有り

    Mohan Amarasiri, Hiroki Kawai, Masaaki Kitajima, Satoshi Okabe, Daisuke Sano

    Water Science and Technology　79　(2)　342-348　2019年1月

    出版者・発行元：{IWA} Publishing

    DOI： 10.2166/wst.2019.050 　

    ISSN：0273-1223

  43. Assays for specific growth rate and cell-binding ability of rotavirus 査読有り

    Syun-suke Kadoya, Daisuke Sano

    Journal of Visualized Experiments　143　e58821　2019年1月

  44. Free-Chlorine Disinfection as a Selection Pressure on Norovirus. 国際誌 査読有り

    Andri Taruna Rachmadi, Masaaki Kitajima, Kozo Watanabe, Sakiko Yaegashi, Joeselle Serrana, Arata Nakamura, Toyoko Nakagomi, Osamu Nakagomi, Kazuhiko Katayama, Satoshi Okabe, Daisuke Sano

    Applied and environmental microbiology　84　(13)　2018年7月1日

    DOI： 10.1128/AEM.00244-18 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    Human noroviruses are excreted in feces from infected individuals and included in wastewater. It is critical to remove/inactivate them in wastewater treatment processes, particularly in the disinfection step, before release to aquatic environments. However, the high mutation rates of human noroviruses raise concerns about the emergence of strains that are less susceptible to disinfectants and can survive even after wastewater treatment. This study aimed to demonstrate the strain-dependent susceptibility of norovirus to free chlorine. A population originated from the murine norovirus strain S7-PP3, a surrogate for human noroviruses in environmental testing, was exposed to free chlorine and then propagated in a host cell. This cycle of free chlorine exposure followed by propagation in cells was repeated 10 times, and populations with lower susceptibility to free chlorine were obtained from two independent trials of chlorine exposure cycles. Open reading frame 2 (ORF2) and ORF3 of the murine norovirus genome were analyzed by next-generation sequencing, and a unique nonsynonymous mutation (corresponding to a change from phenylalanine to serine) at nucleotide (nt) 7280 in ORF3, which encodes the minor capsid protein VP2, was found in chlorine-exposed populations from both trials. It was confirmed that all of the clones from the chlorine-treated population had lower susceptibility to free chlorine than those from the control population. These results indicate that exposure to free chlorine and dilution exert different driving forces to form murine norovirus (MNV) quasispecies, and that there is a selective force to form MNV quasispecies under free chlorine exposure.IMPORTANCE This study showed that free chlorine disinfection exerted a selection pressure for murine norovirus (MNV). The strain-dependent viral susceptibility to the disinfectant elucidated in this study highlights the importance of employing less susceptible strains as representative viruses in disinfection tests, because the disinfection rate values obtained from more susceptible strains would be less useful in predicting the virus inactivation efficiency of circulating strains under practical disinfection conditions.

  45. Virus Particle Detection by Convolutional Neural Network in Transmission Electron Microscopy Images 査読有り

    Eisuke Ito, Takaaki Sato, Daisuke Sano, Etsuko Utagawa, Tsuyoshi Kato

    Food and Environmental Virology　10　(2)　201-208　2018年6月1日

    出版者・発行元：Springer New York LLC

    DOI： 10.1007/s12560-018-9335-7 　

    ISSN：1867-0342 1867-0334

    詳細を見る 詳細を閉じる

    A new computational method for the detection of virus particles in transmission electron microscopy (TEM) images is presented. Our approach is to use a convolutional neural network that transforms a TEM image to a probabilistic map that indicates where virus particles exist in the image. Our proposed approach automatically and simultaneously learns both discriminative features and classifier for virus particle detection by machine learning, in contrast to existing methods that are based on handcrafted features that yield many false positives and require several postprocessing steps. The detection performance of the proposed method was assessed against a dataset of TEM images containing feline calicivirus particles and compared with several existing detection methods, and the state-of-the-art performance of the developed method for detecting virus was demonstrated. Since our method is based on supervised learning that requires both the input images and their corresponding annotations, it is basically used for detection of already-known viruses. However, the method is highly flexible, and the convolutional networks can adapt themselves to any virus particles by learning automatically from an annotated dataset.

  46. A review on recent progress in the detection methods and prevalence of human enteric viruses in water 査読有り

    Eiji Haramoto, Masaaki Kitajima, Akihiko Hata, Jason R. Torrey, Yoshifumi Masago, Daisuke Sano, Hiroyuki Katayama

    Water Research　135　168-186　2018年5月15日

    出版者・発行元：Elsevier Ltd

    DOI： 10.1016/j.watres.2018.02.004 　

    ISSN：1879-2448 0043-1354

    eISSN：1879-2448

    詳細を見る 詳細を閉じる

    Waterborne human enteric viruses, such as noroviruses and adenoviruses, are excreted in the feces of infected individuals and transmitted via the fecal-oral route including contaminated food and water. Since viruses are normally present at low concentrations in aquatic environments, they should be concentrated into smaller volumes prior to downstream molecular biological applications, such as quantitative polymerase chain reaction (qPCR). This review describes recent progress made in the development of concentration and detection methods of human enteric viruses in water, and discusses their applications for providing a better understanding of the prevalence of the viruses in various types of water worldwide. Maximum concentrations of human enteric viruses in water that have been reported in previous studies are summarized to assess viral abundances in aquatic environments. Some descriptions are also available on recent applications of sequencing analyses used to determine the genetic diversity of viral genomes in water samples, including those of novel viruses. Furthermore, the importance and significance of utilizing appropriate process controls during viral analyses are discussed, and three types of process controls are considered: whole process controls, molecular process controls, and (reverse transcription (RT)-)qPCR controls. Although no standards have been established for acceptable values of virus recovery and/or extraction-(RT-)qPCR efficiency, use of at least one of these appropriate control types is highly recommended for more accurate interpretation of observed data.

  47. Microfluidic PCR amplification and MiSeq amplicon sequencing techniques for high-throughput detection and genotyping of human pathogenic RNA viruses in human feces, sewage, and oysters 査読有り

    Mamoru Oshiki, Takayuki Miura, Shinobu Kazama, Takahiro Segawa, Satoshi Ishii, Masashi Hatamoto, Takashi Yamaguchi, Kengo Kubota, Akinori Iguchi, Tadashi Tagawa, Tsutomu Okubo, Shigeki Uemura, Hideki Harada, Naohiro Kobayashi, Nobuo Araki, Daisuke Sano

    Frontiers in Microbiology　9　830　2018年4月27日

    出版者・発行元：Frontiers Media S.A.

    DOI： 10.3389/fmicb.2018.00830 　

    ISSN：1664-302X

    詳細を見る 詳細を閉じる

    Detection and genotyping of pathogenic RNA viruses in human and environmental samples are useful for monitoring the circulation and prevalence of these pathogens, whereas a conventional PCR assay followed by Sanger sequencing is time-consuming and laborious. The present study aimed to develop a high-throughput detection-and-genotyping tool for 11 human RNA viruses [Aichi virus astrovirus enterovirus norovirus genogroup I (GI), GII, and GIV hepatitis A virus hepatitis E virus rotavirus sapovirus and human parechovirus] using a microfluidic device and next-generation sequencer. Microfluidic nested PCR was carried out on a 48.48 Access Array chip, and the amplicons were recovered and used for MiSeq sequencing (Illumina, Tokyo, Japan) genotyping was conducted by homology searching and phylogenetic analysis of the obtained sequence reads. The detection limit of the 11 tested viruses ranged from 100 to 103 copies/μL in cDNA sample, corresponding to 101-104 copies/mL-sewage, 105-108 copies/g-human feces, and 102-105 copies/g-digestive tissues of oyster. The developed assay was successfully applied for simultaneous detection and genotyping of RNA viruses to samples of human feces, sewage, and artificially contaminated oysters. Microfluidic nested PCR followed by MiSeq sequencing enables efficient tracking of the fate of multiple RNA viruses in various environments, which is essential for a better understanding of the circulation of human pathogenic RNA viruses in the human population.

  48. Reverse transcription-quantitative PCR assays for genotype-specific detection of human noroviruses in clinical and environmental samples 査読有り

    Mohan Amarasiri, Masaaki Kitajima, Akiho Miyamura, Ricardo Santos, Silvia Monteiro, Takayuki Miura, Shinobu Kazama, Satoshi Okabe, Daisuke Sano

    International Journal of Hygiene and Environmental Health　221　(3)　578-585　2018年4月

    出版者・発行元：Elsevier {BV}

    DOI： 10.1016/j.ijheh.2018.02.008 　

    ISSN：1438-4639

  49. Selection of cellular genetic markers for the detection of infectious poliovirus 査読有り

    D. Sano, M. Tazawa, M. Inaba, S. Kadoya, R. Watanabe, T. Miura, M. Kitajima, S. Okabe

    Journal of Applied Microbiology　124　(4)　1001-1007　2018年4月1日

    出版者・発行元：Blackwell Publishing Ltd

    DOI： 10.1111/jam.13621 　

    ISSN：1365-2672 1364-5072

    詳細を見る 詳細を閉じる

    Aims: Cellular responses of an established cell line from human intestinal epithelial cells (INT-407 cells) against poliovirus (PV) infections were investigated in order to find cellular genetic markers for infectious PV detection. Methods and Results: Gene expression profile of INT-407 cells was analysed by DNA microarray technique when cells were infected with poliovirus 1 (PV1) (sabin) at multiplicity of infection of 10−3 and incubated for 12 h. Poliovirus infection significantly altered the gene expressions of two ion channels, KCNJ4 and SCN7A. The expression profile of KCNJ4 gene was further investigated by real-time RT-qPCR, and it was found that KCNJ4 gene was significantly regulated at 24 h postinfection of PV1. Conclusions: KCNJ4 gene, coding a potassium channel protein, is proposed as a cellular genetic marker for infectious PV detection. Significance and Impact of the Study: This is the first study to show the availability of cellular responses to detect infectious PV. The selection of cellular genetic markers for infectious viruses using DNA microarray and RT-qPCR can be applicable for the other enteric viruses.

  50. Disinfection as a Selection Pressure on RNA Virus Evolution. 国際誌

    Andri Taruna Rachmadi, Masaaki Kitajima, Kozo Watanabe, Satoshi Okabe, Daisuke Sano

    Environmental science & technology　52　(5)　2434-2435　2018年3月6日

    DOI： 10.1021/acs.est.8b00590 　

    ISSN：0013-936X

  51. Sapovirus in wastewater treatment plants in Tunisia: Prevalence, removal, and genetic characterization 査読有り

    Miguel F. Varela, Imen Ouardani, Tsuyoshi Kato, Syunsuke Kadoya, Mahjoub Aouni, Daisuke Sano, Jesús L. Romalde

    Applied and Environmental Microbiology　84　(6)　e02093-17　2018年3月1日

    出版者・発行元：American Society for Microbiology

    DOI： 10.1128/AEM.02093-17 　

    ISSN：1098-5336 0099-2240

    詳細を見る 詳細を閉じる

    Sapovirus (SaV), from the Caliciviridae family, is a genus of enteric viruses that cause acute gastroenteritis. SaV is shed at high concentrations with feces into wastewater, which is usually discharged into aquatic environments or reused for irrigation without efficient treatments. This study analyzed the incidence of human SaV in four wastewater treatment plants from Tunisia during a period of 13 months (December 2009 to December 2010). Detection and quantification were carried out using reverse transcription-quantitative PCR (RT-qPCR) methods, obtaining a prevalence of 39.9% (87/ 218). Sixty-one positive samples were detected in untreated water and 26 positive samples in processed water. The Dekhila plant presented the highest contamination levels, with a 63.0% prevalence. A dominance of genotype I.2 was observed on 15 of the 24 positive samples that were genetically characterized. By a Bayesian estimation algorithm, the SaV density in wastewater was estimated using left-censored data sets. The mean value of log SaV concentration in untreated wastewater ranged between 2.7 and 4.5 logs. A virus removal efficiency of 0.2 log was calculated for the Dekhila plant as the log ratio posterior distributions between untreated and treated wastewater. Multiple quantitative values obtained in this study must be available in quantitative microbial risk assessment in Tunisia as parameter values reflecting local conditions.

  52. Identification of the inactivating factors and mechanisms exerted on MS2 coliphage in concentrated synthetic urine 査読有り

    Wakana Oishi, Daisuke Sano, Loic Decrey, Syunsuke Kadoya, Tamar Kohn, Naoyuki Funamizu

    Science of the Total Environment　598　213-219　2017年11月

    出版者・発行元：None

    DOI： 10.1016/j.scitotenv.2017.04.088 　

    ISSN：0048-9697

    eISSN：1879-1026

  53. Target virus log(10) reduction values determined for two reclaimed wastewater irrigation scenarios in Japan based on tolerable annual disease burden 査読有り

    Toshihiro Ito, Masaaki Kitajima, Tsuyoshi Kato, Satoshi Ishii, Takahiro Segawa, Satoshi Okabe, Daisuke Sano

    Water Research　125　438-448　2017年11月

    出版者・発行元：None

    DOI： 10.1016/j.watres.2017.08.057 　

    ISSN：0043-1354

  54. Bacteriophage removal efficiency as a validation and operational monitoring tool for virus reduction in wastewater reclamation: Review 査読有り

    Mohan Amarasiri, Masaaki Kitajima, Thanh H. Nguyen, Satoshi Okabe, Daisuke Sano

    Water Research　121　258-269　2017年9月

    出版者・発行元：Elsevier {BV}

    DOI： 10.1016/j.watres.2017.05.035 　

    ISSN：0043-1354

  55. 下水再生処理におけるヒト腸管系ウイルスの目標除去効率の算定法 査読有り

    伊藤 寿宏, 押木 守, 小林 直央, 加藤 毅, 瀬川 高弘, 幡本 将史, 山口 隆司, 原田 秀樹, 北島 正章, 岡部 聡, 佐野 大輔

    環境工学研究論文集　72　(7)　305-313　2017年4月3日

    出版者・発行元：土木学会

    DOI： 10.2208/jscejer.72.III_305 　

  56. Removal of human pathogenic viruses in a down-flow hanging sponge (DHS) reactor treating municipal wastewater and health risks associated with utilization of the effluent for agricultural irrigation 査読有り

    Naohiro Kobayashi, Mamoru Oshiki, Toshihiro Ito, Takahiro Segawa, Masashi Hatamoto, Tsuyoshi Kato, Takashi Yamaguchi, Kengo Kubota, Masanobu Takahashi, Akinori Iguchi, Tadashi Tagawa, Tsutomu Okubo, Shigeki Uemura, Hideki Harada, Toshiki Motoyama, Nobuo Araki, Daisuke Sano

    Water Research　110　389-398　2017年3月

    出版者・発行元：None

    DOI： 10.1016/j.watres.2016.10.054 　

    ISSN：0043-1354

  57. Evaluation of virus reduction efficiency in wastewater treatment unit processes as a credit value in the multiple-barrier system for wastewater reclamation and reuse 査読有り

    Toshihiro Ito, Tsuyoshi Kato, Makoto Hasegawa, Hiroyuki Katayama, Satoshi Ishii, Satoshi Okabe, Daisuke Sano

    Journal of Water and Health　14　(6)　879-889　2016年12月

    出版者・発行元：None

    DOI： 10.2166/wh.2016.096 　

    ISSN：1477-8920

  58. Bactericidal and virucidal mechanisms in the alkaline disinfection of compost using calcium lime and ash 査読有り

    Nowaki Hijikata, Rui Tezuka, Shinobu Kazama, Masahiro Otaki, Ken Ushijima, Ryusei Ito, Satoshi Okabe, Daisuke Sano, Naoyuki Funamizu

    Journal of Environmental Management　181　721-727　2016年10月

    出版者・発行元：None

    DOI： 10.1016/j.jenvman.2016.08.026 　

    ISSN：0301-4797

    eISSN：1095-8630

  59. Effect of leaf surface chemical properties on efficacy of sanitizer for rotavirus inactivation 査読有り

    Miyu Fuzawa, Kang-Mo Ku, Sindy Paola Palma-Salgado, Kenya Nagasaka, Hao Feng, John A. Juvik, Daisuke Sano, Joanna L. Shisler, Thanh H. Nguyen

    Applied and Environmental Microbiology　82　(20)　6214-6222　2016年10月

    出版者・発行元：None

    DOI： 10.1128/AEM.01778-16 　

    ISSN：0099-2240

    eISSN：1098-5336

  60. Risk management of viral infectious diseases in wastewater reclamation and reuse: Review 査読有り

    Daisuke Sano, Mohan Amarasiri, Akihiko Hata, Toru Watanabe, Hiroyuki Katayama

    Environment International　91　220-229　2016年5月

    出版者・発行元：None

    DOI： 10.1016/j.envint.2016.03.001 　

    ISSN：0160-4120

  61. Bacterial histo-blood group antigens contributing to genotype-dependent removal of human noroviruses with a microfiltration membrane 査読有り

    Mohan Amarasiri, Satoshi Hashiba, Takayuki Miura, Toyoko Nakagomi, Osamu Nakagomi, Satoshi Ishii, Satoshi Okabe, Daisuke Sano

    Water Research　95　383-391　2016年5月

    出版者・発行元：Elsevier {BV}

    DOI： 10.1016/j.watres.2016.04.018 　

    ISSN：0043-1354

  62. 水中病原体の許容感染リスクに基づいた水質衛生基準候補値の算出方法に関する提案 査読有り

    小林彩乃, 佐野大輔, 加藤毅, 伊藤寿宏, 宮村明帆, 三浦尚之, 石井聡, 岡部聡

    土木学会論文集　72　(3)　40-49　2016年3月

    出版者・発行元：公益社団法人 土木学会

    DOI： 10.2208/jscejer.72.40 　

    詳細を見る 詳細を閉じる

    本研究では環境水の水質衛生基準候補値を算出する方法を提案する．まずモニタリング調査結果をもとに指標微生物に対する病原体の濃度比分布を推定し，さらに定量的微生物リスク評価の手法を用いることで仮定された水利用形態における病原体の河川水中許容濃度を算出した．この病原体の水中許容濃度は，指標微生物の病原体に対する濃度比分布と照合され，安全率を勘案した上で水質衛生基準値としての指標微生物濃度を算出した．個体差に関わる安全率を10とした場合，生活環境の保全に関する環境基準（河川）B類型における大腸菌の水質衛生基準値の候補として150 MPN/ 100mLを得た．本研究で提案した手法により，モニタリングデータの活用と仮定の検証を進めることで，「科学的根拠」に基づいた水質衛生基準値の設定が可能となる．

  63. Estimation of concentration ratio of indicator to pathogen-related gene in environmental water based on left-censored data 査読有り

    Tsuyoshi Kato, Ayano Kobayashi, Toshihiro Ito, Takayuki Miura, Satoshi Ishii, Satoshi Okabe, Daisuke Sano

    Journal of Water and Health　14　(1)　14-25　2016年2月

    出版者・発行元：None

    DOI： 10.2166/wh.2015.029 　

    ISSN：1477-8920

  64. 環境と胃腸炎ウイルス：ウイルス吸着性細菌との関わり

    佐野 大輔

    ウイルス　66　(2)　179-186　2016年

    出版者・発行元：日本ウイルス学会

    DOI： 10.2222/jsv.66.179 　

    ISSN：0042-6857

    詳細を見る 詳細を閉じる

    ノロウイルスやロタウイルスに代表される胃腸炎ウイルスは，ヒト-ヒト間の感染に加えて，水や食品，もしくはドアノブなどの環境表面を介して感染するため，感染制御のためには感染者との接触を避けるだけでなく，胃腸炎ウイルスに汚染された水，食品及び環境表面の適切な消毒が求められる．本稿の著者らは，これまでの研究において，胃腸炎ウイルスの消毒剤感受性や水環境中動態に影響を与えうる存在として，血液型決定抗原（Histo-blood group antigen: HBGA）陽性細菌に着目してきた．本稿では，HBGA陽性細菌に関わる研究事例をまとめ，胃腸炎ウイルスの生活環に与える影響について，これまでに報告されている内容を紹介する．

  65. Genome sequence of Enterobacter cloacae strain SENG-6, a bacterium producing histo-blood group antigen-like substances that can bind with human noroviruses

    Satoshi Ishii, Mohan Amarasiri, Satoshi Hashiba, Peiyi Yang, Satoshi Okabe, Daisuke Sano

    Genome Announcements　4　(4)　e00893-16　2016年

    出版者・発行元：American Society for Microbiology

    DOI： 10.1128/genomeA.00893-16 　

  66. Bayesian modeling of virus removal efficiency in wastewater treatment processes 査読有り

    T. Ito, T. Kato, K. Takagishi, S. Okabe, D. Sano

    Water Science and Technology　72　(10)　1789-1795　2015年11月

    出版者・発行元：None

    DOI： 10.2166/wst.2015.402 　

    ISSN：0273-1223

    eISSN：1996-9732

  67. Removal properties of human enteric viruses in a pilot-scale membrane bioreactor (MBR) process 査読有り

    Takayuki Miura, Satoshi Okabe, Yoshihito Nakahara, Daisuke Sano

    Water Research　75　282-291　2015年5月

    出版者・発行元：None

    DOI： 10.1016/j.watres.2015.02.046 　

    ISSN：0043-1354

  68. Culture-independent evaluation of nonenveloped-virus infectivity reduced by free-chlorine disinfection 査読有り

    Daisuke Sano, Takatomo Ohta, Arata Nakamura, Toyoko Nakagomi, Osamu Nakagomi, Satoshi Okabe

    Applied and Environmental Microbiology　81　(8)　2819-2826　2015年4月

    出版者・発行元：None

    DOI： 10.1128/AEM.03802-14 　

    ISSN：0099-2240

    eISSN：1098-5336

  69. Molecular epidemiology of noroviruses detected in Nepalese children with acute diarrhea between 2005 and 2011: Increase and predominance of minor genotype GII.13 査読有り

    Thi Nguyen Hoa-Tran, Toyoko Nakagomi, Daisuke Sano, Jeevan B. Sherchand, Basu D. Pandey, Nigel A. Cunliffe, Osamu Nakagomi

    Infection Genetics and Evolution　30　27-36　2015年3月

    出版者・発行元：None

    DOI： 10.1016/j.meegid.2014.12.003 　

    ISSN：1567-1348

    eISSN：1567-7257

  70. 衛生環境工学分野におけるマイクロビオーム研究 招待有り

    井原賢, 稲葉愛美, 佐野大輔

    臨床と微生物　42　(6)　685-689　2015年

    ISSN：0910-7029

  71. 将来の社会システムにおける感染性胃腸炎リスク 招待有り

    佐野 大輔

    最新医学　70　2415-2422　2015年

  72. 水中の健康関連微生物に関する研究動向

    片山浩之, 安井宣仁, 猪又明子, 田中宏明, 岸田直裕, 原本英司, 渡部徹, 真砂佳史, 浅田安廣, 端昭彦, 佐野大輔, 井原賢, 稲葉愛美

    水環境学会誌　37(A)　(12)　424-433　2014年12月10日

    ISSN：0916-8958

  73. Microfluidic quantitative PCR for simultaneous quantification of multiple viruses in environmental water samples 査読有り

    Satoshi Ishii, Gaku Kitamura, Takahiro Segawa, Ayano Kobayashi, Takayuki Miura, Daisuke Sano, Satoshi Okabe

    Applied and Environmental Microbiology　80　(24)　7505-7511　2014年12月

    出版者・発行元：None

    DOI： 10.1128/AEM.02578-14 　

    ISSN：0099-2240

    eISSN：1098-5336

  74. External CO2 and water supplies for enhancing electrical power generation of air-cathode microbial fuel cells 査読有り

    So Ishizaki, Itto Fujiki, Daisuke Sano, Satoshi Okabe

    Environmental Science & Technology　48　(19)　11204-11210　2014年10月

    出版者・発行元：None

    DOI： 10.1021/es5021197 　

    ISSN：0013-936X

    eISSN：1520-5851

  75. Estimation of contamination sources of human enteroviruses in a wastewater treatment and reclamation system by PCR-DGGE 査読有り

    Zheng Ji, Xiaochang C. Wang, Limei Xu, Chongmiao Zhang, Naoyuki Funamizu, Satoshi Okabe, Daisuke Sano

    Food and Environmental Virology　6　(2)　99-109　2014年6月

    出版者・発行元：None

    DOI： 10.1007/s12560-014-9140-x 　

    ISSN：1867-0334

    eISSN：1867-0342

  76. Water quality monitoring and risk assessment by simultaneous multipathogen quantification 査読有り

    Satoshi Ishii, Takamitsu Nakamura, Shuji Ozawa, Ayano Kobayashi, Daisuke Sano, Satoshi Okabe

    Environmental Science & Technology　48　(9)　4744-4749　2014年5月

    出版者・発行元：None

    DOI： 10.1021/es500578s 　

    ISSN：0013-936X

    eISSN：1520-5851

  77. 水環境中からのウイルス情報の収集とその活用・総説 招待有り

    佐野 大輔

    臨床とウイルス　42　(5)　211-223　2014年

  78. 培養できないウイルスの感染性の評価法 招待有り

    佐野大輔, 岡部聡

    感染と消毒　21　(2)　24-27　2014年

  79. Bayesian modeling of enteric virus density in wastewater using left-censored data 査読有り

    Tsuyoshi Kato, Takayuki Miura, Satoshi Okabe, Daisuke Sano

    Food and Environmental Virology　5　(4)　185-193　2013年12月

    出版者・発行元：None

    DOI： 10.1007/s12560-013-9125-1 　

    ISSN：1867-0334

    eISSN：1867-0342

  80. Use of a genetically-engineered Escherichia coli strain as a sample process control for quantification of the host-specific bacterial genetic markers 査読有り

    Ayano Kobayashi, Daisuke Sano, Asami Taniuchi, Satoshi Ishii, Satoshi Okabe

    Applied and Environmental Microbiology　97　(20)　9165-9173　2013年10月

    出版者・発行元：None

    DOI： 10.1007/s00253-013-5188-z 　

    ISSN：0175-7598

  81. Histo-blood group antigen-like substances of human enteric bacteria as specific adsorbents for human noroviruses 査読有り

    Takayuki Miura, Daisuke Sano, Atsushi Suenaga, Takeshi Yoshimura, Miyu Fuzawa, Toyoko Nakagomi, Osamu Nakagomi, Satoshi Okabe

    Journal of Virology　87　(17)　9441-9451　2013年9月

    出版者・発行元：None

    DOI： 10.1128/JVI.01060-13 　

    ISSN：0022-538X

  82. Chicken- and duck-associated Bacteroides-Prevotella genetic markers for detecting fecal contamination in environmental water 査読有り

    Ayano Kobayashi, Daisuke Sano, Jun Hatori, Satoshi Ishii, Satoshi Okabe

    Applied Microbiology and Biotechnology　97　(16)　7427-7437　2013年8月

    出版者・発行元：None

    DOI： 10.1007/s00253-012-4469-2 　

    ISSN：0175-7598

  83. 水環境中の病原ウイルス分析手法

    三浦尚之, 佐野大輔

    日本水環境学会誌　5　151-155　2013年

  84. 生活用水に潜むウイルス対策 招待有り

    佐野大輔

    バムサジャーナル　25　(4)　8-13　2013年

  85. A new approach for evaluating the infectivity of noncultivatable enteric viruses without cell culture 査読有り

    Kazuki Tojo, Daisuke Sano, Takayuki Miura, Toyoko Nakagomi, Osamu Nakagomi, Satoshi Okabe

    Water Science and Technology　67　(10)　2236-2240　2013年

    出版者・発行元：None

    DOI： 10.2166/wst.2013.114 　

    ISSN：0273-1223

    eISSN：1996-9732

  86. Effects of temperature and predator on the persistence of host-specific Bacteroides-Prevotella genetic markers in water 査読有り

    Ayano Kobayashi, Daisuke Sano, Satoshi Okabe

    Water Science and Technology　67　(4)　838-845　2013年

    出版者・発行元：4

    DOI： 10.2166/wst.2012.626 　

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Genetic markers derived from Bacteroidales spp. have been proposed as promising indicators for fecal contamination in the water environment. However, little is known about the persistency of Bacteroidales spp. 16S rRNA genetic markers in the natural environment, which hampers the precise identification of fecal contamination sources. In this study, the persistency of human-specific Bacteroidales spp. genetic markers in river water was investigated during a 3-week agitation. The copy number of Bacteroidales spp. genetic marker was decreased with agitation time, and was very sensitive to water temperature. After the 3-week agitation, three clones of 18S rRNA gene related to Glaucoma scintillans, Spumella-like flagellate, and Colpidium campylum were acquired. The presence of predators that can prey on target bacteria could also be a critical factor affecting the quantified value of genetic markers. It is very important to take these factors, water temperature and the presence of predator, into account for predicting the fate of genetic markers to accurately identify fecal pollution sources. © IWA Publishing 2013.

  87. Occurrence of Hand-Foot-and-Mouth Disease Pathogens in Domestic Sewage and Secondary Effluent in Xi'an, China 査読有り

    Zheng Ji, Xiaochang Wang, Chongmiao Zhang, Takayuki Miura, Daisuke Sano, Naoyuki Funamizu, Satoshi Okabe

    MICROBES AND ENVIRONMENTS　27　(3)　288-292　2012年9月

    出版者・発行元：JAPANESE SOC MICROBIAL ECOLOGY, DEPT BIORESOURCE SCIENCE

    DOI： 10.1264/jsme2.ME11352 　

    ISSN：1342-6311

    詳細を見る 詳細を閉じる

    Hand, foot and mouth disease (HFMD), caused by a group of enteric viruses such as Enterovirus 71 (EV71), Coxsackievirus A16 (CVA16) and Coxsackievirus A10 (CVA10), is heavily epidemic in East Asia. This research focused on investigating the occurrence of HFMD pathogens in domestic sewage and secondary effluent before disinfection in a wastewater treatment plant (WWTP) in Xi'an, the largest megacity in northwest China. In order to simultaneously detect all three HFMD pathogens, a semi-nested RT-PCR assay was constructed with a newly designed primer set targeting conservative gene regions from the 5' untranslated region (UTR) to VP2. As a result, 86% of raw sewage samples and 29% of the secondary effluent samples were positive for the HFMD viral gene, indicating that HFMD pathogens were highly prevalent in domestic wastewater and that they could also persist, even with lower probability, in the secondary effluent before disinfection. Of the three HFMD pathogens, CVA10 was positive in 48% of the total samples, while the occurrences of CVA16 and EV71 were 12% and 2%, respectively. It could thus be stated that CVA10 is the main HFMD pathogen prevailing in the study area, at least during the investigation period. High genetic diversity in the conservative gene region among the same serotype of the HFMD pathogen was identified by phylogenetic analysis, implying that this HFMD pathogen replicates frequently among the population excreting the domestic sewage.

  88. Human norovirus occurrence and diversity in the Llobregat river catchment, Spain 査読有り

    Unai Perez-Sautu, Daisuke Sano, Susana Guix, Georg Kasimir, Rosa M. Pinto, Albert Bosch

    ENVIRONMENTAL MICROBIOLOGY　14　(2)　494-502　2012年2月

    出版者・発行元：WILEY-BLACKWELL

    DOI： 10.1111/j.1462-2920.2011.02642.x 　

    ISSN：1462-2912

    詳細を見る 詳細を閉じる

    Human noroviruses (NoV) were quantified and characterized in an 18 month survey conducted along the Llobregat river catchment in Spain. Sample types included freshwater, untreated and treated wastewater and drinking water. High NoV genome copy numbers were reported, reaching up to 10(6) l(-1) and 10(9) l(-1) in freshwater and raw sewage respectively. In both types of samples, GII NoV genome copies outnumbered those of GI, although without significance. All samples of semi-treated and treated drinking water were negative for NoV. A clear seasonality of NoV occurrence was observed both in river water and sewage samples, with significantly higher genome copy numbers in the cold than in the warm months period. Mean NoV log reduction rates after biological treatment of sewage were 2.2 and 3.1 for GI and GII respectively. A total of 77 NoV strains isolated in the Llobregat river catchment could be phylogenetically characterized, 44 belonging to GI and 33 to GII. The most prevalent genotype was GI. 4, followed by GII.4 and GII.21. Several variants of the pandemic GII.4 strain were detected in the environment, corroborating their circulation among the population.

  89. Adsorption characteristics of an enteric virus-binding protein to norovirus, rotavirus and poliovirus 査読有り

    Takahiro Imai, Daisuke Sano, Takayuki Miura, Satoshi Okabe, Keishi Wada, Yoshifumi Masago, Tatsuo Omura

    BMC Biotechnology　11　2011年12月16日

    DOI： 10.1186/1472-6750-11-123 　

    ISSN：1472-6750

    詳細を見る 詳細を閉じる

    Background: Water contamination with human enteric viruses has posed human health risks all over the world. Reasonable and facile methodologies for recovering and quantifying infectious enteric viruses in environmental samples are needed to address the issues of waterborne viral infectious diseases. In this study, a bacterial protein that has a binding capability with several enteric viruses is discovered, and its binding characteristics were investigated for utilizing it as a viral adsorbent in virus recovery and detection technologies.Results: A gene of an enteric virus-binding protein (EVBP), derived from a monomer of a bacterial chaperon protein GroEL, was successfully acquired from a genomic DNA library of activated sludge microorganisms with nested PCR. Equilibrium dissociation constants between EVBP and norovirus-like particles (NoVLPs) of genotypes GI.7 and GII.4, estimated with quartz crystal microbalance method, were 240 and 210 nM, respectively. These values of equilibrium dissociation constant imply that the binding affinity between EVBP and NoVLPs is 1 to 3-log weaker than that in general antigen-antibody interactions, but about 2-log stronger than that in weak specific interactions of proteins with cations and organic polymers. The adsorptions of EVBP to norovirus, group A rotavirus and poliovirus type 1 were found to be significant in enzyme-linked immunosorbent assay. Meanwhile, the binding of native GroEL tetradecamer to viral particles was weaker than that of EVBP, presumably because of a steric hindrance. The small molecule of EVBP could have an advantage in the access to the surface of viral particles with rugged structure.Conclusions: EVBP that has a broad binding spectrum to enteric viruses was newly discovered. The broad binding characteristic of EVBP would allow us to utilize it as a novel adsorbent for detecting diverse enteric viruses in clinical and environmental samples. © 2011 Imai et al licensee BioMed Central Ltd.

  90. Development of an Effective Method for Recovery of Viral Genomic RNA from Environmental Silty Sediments for Quantitative Molecular Detection 査読有り

    Takayuki Miura, Yoshifumi Masago, Daisuke Sano, Tatsuo Omura

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　77　(12)　3975-3981　2011年6月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.02692-10 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    Nine approaches to recover viral RNA from environmental silty sediments were newly developed and compared to quantify RNA viruses in sediments using molecular methods. Four of the nine approaches employed direct procedures for extracting RNA from sediments (direct methods), and the remaining five approaches used indirect methods wherein viral particles were recovered before RNA extraction. A direct method using an SDS buffer with EDTA to lyse viral capsids in sediments, phenol-chloroform-isoamyl alcohol to extract RNA, isopropanol to concentrate RNA, and magnetic beads to purify RNA resulted in the highest rate of recovery (geometric mean of 11%, with a geometric standard deviation of 0.02; n = 7) of poliovirus 1 (PV1) inoculated in an environmental sediment sample. The direct method exhibiting the highest rate of PV1 recovery was applied to environmental sediment samples. One hundred eight sediment samples were collected from the Takagi River, Miyagi, Japan, and its estuary from November 2007 to April 2009, and the genomic RNAs of enterovirus and human norovirus in these samples were quantified by reverse transcription (RT)-quantitative PCR (qPCR). The human norovirus genome was detected in one sample collected at the bay, although its concentration was below the quantification limit. Meanwhile, the enterovirus genome was detected in two samples at the river mouth and river at concentrations of 8.6 x 10(2) and 2.4 x 10(2) copies/g (wet weight), respectively. This is the first report to obtain quantitative data for a human pathogenic virus in a river and in estuarine sediments using RT-qPCR.

  91. Development and validation of a microarray for the confirmation and typing of norovirus RT-PCR products 査読有り

    Kirsten Mattison, Nathalie Corneau, Ingvild Berg, Albert Bosch, Erwin Duizer, Ion Gutierrez-Aguirre, Yvan L&apos;Homme, Yalda Lucero, Zhiyao Luo, Alexander Martyres, Mette Myrmel, Miguel O&apos;Ryan, Franco Pagotto, Daisuke Sano, Sanela Svraka, Ulises Urzua, Sabah Bidawid

    JOURNAL OF VIROLOGICAL METHODS　173　(2)　233-250　2011年5月

    出版者・発行元：ELSEVIER SCIENCE BV

    DOI： 10.1016/j.jviromet.2011.02.011 　

    ISSN：0166-0934

    詳細を見る 詳細を閉じる

    Noroviruses are implicated in many worldwide institutional, food and waterborne outbreaks each year. Genetic typing of isolates is valuable for monitoring outbreak spread as well as variation in circulating strains. Microarrays have the potential to provide rapid genotype information for norovirus samples. The NoroChip v3.0 provides an oligonucleotide hybridization platform to screen for over 600 potential interactions in each experiment. The NoroChip v3.0 was developed at Health Canada and validated in seven international partner laboratories. Each laboratory validated the NoroChip v3.0 using norovirus amplicons routinely characterized in their testing protocols. Fragments from the capsid region (region C) and a 2.4 kb amplicon spanning polymerase and capsid sequences (region AD) were validated in six of the partner laboratories and provided correct genogroup typing information (GI or GII) when hybridized to the NoroChip v3.0. Results indicate that the current limiting factor for implementing the NoroChip v3.0 as a strain typing tool is the difficulty obtaining a long, specific amplicon from all circulating norovirus strains. Data obtained with the longer region AD amplicon provided the best discrimination between norovirus strains. Crown Copyright (C) 2011 Published by Elsevier B.V. All rights reserved.

  92. 感染性胃腸炎の診断,治療,伝播予防 7.感染性胃腸炎と上下水道の衛生工学的管理

    佐野大輔

    化学療法の領域　27　(4)　734-740　2011年3月

    ISSN：0913-2384

  93. Analytical Methods for Virus Detection in Water and Food 査読有り

    Albert Bosch, Gloria Sanchez, Morteza Abbaszadegan, Annalaura Carducci, Susana Guix, Francoise S. Le Guyader, Rembuluwani Netshikweta, Rosa M. Pinto, Wim H. M. van der Poel, Saskia Rutjes, Daisuke Sano, Maureen B. Taylor, Walda B. van Zyl, David Rodriguez-Lazaro, Katarina Kovac, Jane Sellwood

    FOOD ANALYTICAL METHODS　4　(1)　4-12　2011年3月

    出版者・発行元：SPRINGER

    DOI： 10.1007/s12161-010-9161-5 　

    ISSN：1936-9751

    eISSN：1936-976X

    詳細を見る 詳細を閉じる

    Potential ways to address the issues that relate to the techniques for analyzing food and environmental samples for the presence of enteric viruses are discussed. It is not the authors' remit to produce or recommend standard or reference methods but to address specific issues in the analytical procedures. Foods of primary importance are bivalve molluscs, particularly, oysters, clams, and mussels; salad crops such as lettuce, green onions and other greens; and soft fruits such as raspberries and strawberries. All types of water, not only drinking water but also recreational water (fresh, marine, and swimming pool), river water (irrigation water), raw and treated sewage are potential vehicles for virus transmission. Well over 100 different enteric viruses could be food or water contaminants; however, with few exceptions, most well-characterized foodborne or waterborne viral outbreaks are restricted to hepatitis A virus (HAV) and calicivirus, essentially norovirus (NoV). Target viruses for analytical methods include, in addition to NoV and HAV, hepatitis E virus (HEV), enteroviruses (e.g., poliovirus), adenovirus, rotavirus, astrovirus, and any other relevant virus likely to be transmitted by food or water. A survey of the currently available methods for detection of viruses in food and environmental matrices was conducted, gathering information on protocols for extraction of viruses from various matrices and on the various specific detection techniques for each virus type.

  94. Quantification and Genotyping of Human Sapoviruses in the Llobregat River Catchment, Spain 査読有り

    Daisuke Sano, Unai Perez-Sautu, Susana Guix, Rosa Maria Pinto, Takayuki Miura, Satoshi Okabe, Albert Bosch

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　77　(3)　1111-1114　2011年2月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.01721-10 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    Human sapoviruses (SaVs) were quantified and characterized in an 18-month survey conducted along the Llobregat river catchment area in Spain. Sample types included freshwater, untreated and treated wastewater, and drinking water. All genogroups were recovered, and a seasonal distribution was observed. This is the first report of SaV quantification and genotyping in the environment outside Japan.

  95. Identification and characterization of coagulation inhibitor proteins derived from cyanobacterium Microcystis aeruginosa. 査読有り

    Sano, Daisuke, Ishifuji, Shingo, Sato, Yuichi, Imae, Yasutaka, Takaara, Tomoko, Masago, Yoshifumi, Omura, Tatsuo

    Chemosphere　82　(8)　1096-1102　2011年

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1016/j.chemosphere.2010.12.005 　

    ISSN：0045-6535

    詳細を見る 詳細を閉じる

    The excess growth of cyanobacteria in semi enclosed water areas caused by eutrophication brings about coagulation inhibition in drinking water treatment processes. In this study, coagulation inhibitor proteins produced by Microcystis aeruginosa, a major cyanobacterium in algal bloom, were acquired by a phage display technique, an aluminum-immobilized affinity chromatography and a protein expression technique using Escherichia coli cells. Two cyanobacterial peptides with a high ratio of metallophilic amino acids were recovered, which were a part of homologues of a thiol oxidase enzyme Ero1p and a trans-acting repressor ArsR. It was also shown that the homologue of ArsR exhibited a stronger inhibitory effect on the coagulation of kaolin suspension with polyaluminum chloride than the control proteins. This is the first report to identify a cyanobacterial cell component to inhibit coagulation. The compositions of polar amino acids were critical to explain the strength of coagulation inhibition potential. Polar proteins from cyanobacteria could collectively consume coagulants or stabilize clay particles, which would be plausible explanations for causing coagulation inhibition. Meanwhile, results from the kaolin coagulation tests using the control proteins implied that the neutralization of positive charges of coagulant constituents by simple electrostatic interactions might not be the key mechanism on the protein-induced coagulation inhibition. (C) 2010 Elsevier Ltd. All rights reserved.

  96. Norovirus-binding proteins recovered from activated sludge micro-organisms with an affinity to a noroviral capsid peptide 査読有り

    D. Sano, K. Wada, T. Imai, Y. Masago, T. Omura

    JOURNAL OF APPLIED MICROBIOLOGY　109　(6)　1923-1928　2010年12月

    出版者・発行元：WILEY-BLACKWELL PUBLISHING, INC

    DOI： 10.1111/j.1365-2672.2010.04821.x 　

    ISSN：1364-5072

    詳細を見る 詳細を閉じる

    Aims: Transmission routes of noroviruses, leading aetiological agents of acute gastroenteritis, are rarely verified when outbreaks occur. Because the destination of norovirus particles being firmly captured by micro-organisms could be totally different from that of those particles moving freely, micro-organisms with natural affinity ligands such as virus-binding proteins would affect the fate of viruses in environment, if such microbial affinity ligands exist. The aim of this study is to identify norovirus-binding proteins (NoVBPs) that are presumably working as natural ligands for norovirus particles in water environments. Methods and Results: NoVBPs were recovered from activated sludge microorganisms by an affinity chromatography technique in which a capsid peptide of norovirus genogroup II (GII) was immobilized. The recovered NoVBPs bind to norovirus-like particles (NoVLPs) of norovirus GII, and this adsorption was stronger than that to NoVLPs of norovirus genogroup I. The profile of two-dimensional electrophoresis of NoVBPs showed that the recovered NoVBPs included at least seven spots of protein. The determination of N-terminal amino acid sequences of these NoVBPs revealed that hydrophobic interactions could contribute to the adsorption between NoVBPs and norovirus particles. Conclusions: NoVBPs conferring a high affinity to norovirus GII were successfully isolated from activated sludge micro-organisms. Significance and Impact of the Study: NoVBPs could be natural viral ligands and play an important role in the NoV transmission.

  97. ノロウイルス制御―院内伝播防止に向けて 2)水系管理によるノロウイルス感染症の制御の可能性

    佐野大輔

    感染対策ICTジャーナル　5　(4)　421-426　2010年10月

    ISSN：1881-4964

  98. Detection of Sapovirus in oysters 査読有り

    You Ueki, Mika Shoji, Yoko Okimura, Yasuko Miyota, Yoshifumi Masago, Tomoichiro Oka, Kazuhiko Katayama, Naokazu Takeda, Mamoru Noda, Takayuki Miura, Daisuke Sano, Tatsuo Omura

    MICROBIOLOGY AND IMMUNOLOGY　54　(8)　483-486　2010年8月

    出版者・発行元：WILEY-BLACKWELL

    DOI： 10.1111/j.1348-0421.2010.00239.x 　

    ISSN：0385-5600

    詳細を見る 詳細を閉じる

    SaV sequences which are either genetically identical or similar were detected from oysters, feces from gastroenteritis patients, and domestic wastewater samples in geographically close areas. This is the first report of the detection of SaV in oysters which meet the legal requirements for raw consumption in Japan.

  99. Surface-retained organic matter of Microcystis aeruginosa inhibiting coagulation with polyaluminum chloride in drinking water treatment 査読有り

    Tomoko Takaara, Daisuke Sano, Yoshifumi Masago, Tatsuo Omura

    WATER RESEARCH　44　(13)　3781-3786　2010年7月

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1016/j.watres.2010.04.030 　

    ISSN：0043-1354

    詳細を見る 詳細を閉じる

    Algogenic organic matter produced by the excess growth of cyanobacteria in semi-closed water areas causes coagulation inhibition in drinking water production. In this study, hydrophilic substances of Microcystis aeruginosa, which were mainly composed of lipopolysaccharide (LPS) and RNA, were prepared, and the involvement of these cyanobacterial hydrophilic substances in coagulation inhibition was investigated. As a result, it was found that the negatively charged hydrophilic substances with a molecular weight higher than 10 kDa have a significant role in coagulation inhibition. Further fractionation of cyanobacterial hydrophilic substances revealed that surface-retained organic matter (SOM), including LPS, could exhibit a potent inhibitory effect on the coagulation using polyaluminum chloride (PACl), presumably because of the direct interaction of hydrophilic SOM with cations originated from PACl, which could impede the hydrolysis of the coagulant. (C) 2010 Elsevier Ltd. All rights reserved.

  100. Microcystis aeruginosa由来凝集阻害誘因タンパク質の同定 査読有り

    石藤慎吾, 佐藤有一, 今江泰貴, 高荒智子, 佐野大輔, 真砂佳史, 大村達夫

    水環境学会誌　33　(6)　73-79　2010年6月

    出版者・発行元：Japan Society on Water Environment

    DOI： 10.2965/jswe.33.73 　

    ISSN：0916-8958

    詳細を見る 詳細を閉じる

    Excess growth of cyanobacteria in semiclosed water area caused by eutrophication induces coagulation inhibition in drinking water treatment processes. Thus, it is highly desirable to establish effective countermeasures against such coagulation inhibition by identifying the causative substances released by cyanobacteria. In this study, coagulation inhibitor proteins of <i>Microcystis aeruginosa</i> were screened using a phage display system and an aluminum-ion-immobilized affinity chromatography. From the results, algal proteins with a high level of metal-coordinating amino acids were identified as inhibitor proteins for the coagulation. One of the inhibitor proteins, a homologue of the metal sensor protein ArsR, showed strong inhibition of the coagulation of kaolin suspension with polyaluminum chloride (PAC). It was demonstrated that one of the mechanisms of the coagulation inhibition caused by <i>M. aeruginosa</i> is the inhibition of the cross-linking between aluminum clusters in PAC and suspended solids by the algal inhibitor protein.

  101. Detection of Oxidative Damages on Viral Capsid Protein for Evaluating Structural Integrity and Infectivity of Human Norovirus 査読有り

    Daisuke Sano, Rosa M. Pinto, Tatsuo Omura, Albert Bosch

    ENVIRONMENTAL SCIENCE & TECHNOLOGY　44　(2)　808-812　2010年1月

    出版者・発行元：AMER CHEMICAL SOC

    DOI： 10.1021/es9018964 　

    ISSN：0013-936X

    詳細を見る 詳細を閉じる

    The infectivity evaluation of noncultivatable viruses, such as human norovirus, is crucial to address needs for ensuring the safety in usage of water and marine products. In this work, we tested a new approach to Evaluate viral particle integrity, in which oxidatively produced carbonyl groups on viral capsid protein were quantitatively detected. As a result the decrease in the infectivity of human astrovirus, a representative enteric virus, positively correlated with the amount of oxidative damage on viral particles. Furthermore, when human norovirus was treated by 1 ppm free chlorine for 15 min, 49.93% of virions were recovered as oxidatively damaged particles, which represents a 5-fold increase over those treated by 0.5 ppm free chlorine for 15 min. The detection of the carbonylated viral particles could be a powerful tool for the evaluation of the decrease in the infectivity of noncultivatable viruses.

  102. 水環境及び感染性胃腸炎患者から得られたノロウイルスカプシドタンパク質遺伝子の多様性及びアミノ酸配列変異の解析 査読有り

    今井崇博, 佐野大輔, 真砂佳史, 植木洋, 福士謙介, 大村達夫

    環境工学研究論文集　45　(45)　355-360　2008年11月

    出版者・発行元：公益社団法人 土木学会

    DOI： 10.11532/proes1992.45.355 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    河川水やカキ等の水環境由来サンプルと感染性胃腸炎患者由来糞便を同一地域において約4年間に渡って収集し, ノロウイルスカプシドタンパク質遺伝子を検出, 解読した.さらに解読された塩基配列にコードされたアミノ酸配列における変異に着目し, その多様性を評価した.得られたノロウイルスカプシドタンパク質遺伝子配列は, GII.2 (7配列), GII.3 (25配列), GII.4 (41配列), GII.5 (5配列), GII.6 (1配列), GII.10 (3配列), GII.11 (2配列), GII.14 (10配列) 及びGII.15 (1配列) の各遺伝子型に分類された.このうち世界中で感染性胃腸炎を引き起こしている遺伝子型であるノロウイルスGII.4のカプシドタンパク質遺伝子にコードされたアミノ酸配列について, 本研究で得られた配列をプロトタイプ (Bristol株) と比較したところ, 第8残基のアラニン, 第9残基のアスパラギン, 第15残基のアラニン及び第54残基のアスパラギンにおいてアミノ酸置換が確認された.保存性の高い領域の遺伝子配列だけでなく, そこにコードされたアミノ酸配列に着目することで, 人間社会におけるノロウイルスの動態を探る上で追跡すべき重要な遺伝子領域を絞り込むことが可能となった.

  103. Enzymatic Virus Elution法によるカキ中腸腺からのウイルス誘出技術の開発 査読有り

    奥村千恵, 真砂佳史, 佐野大輔, 植木洋, 大村達夫

    環境工学研究論文集　45　179-186　2008年11月

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.45.179 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    本研究では, 酵素を用いたウイルス誘出手法 (Enzumatic Virus Elution法) よにる, カキ中腸腺からの新しいウイルス 誘出技術を開発した. 中腸腺試料にポリオウイルスを添加し, 3種類の酵素を用いた処理を試みた結果, 脂質分解酵 素であるリパーゼで処理することで, 酵素を使用しない場合と比較して検出効率が0.13%から9.6%へ74倍に改善された. ウイルス検出の各操作過程の中では, ウイルス誘出操作でのウイルス粒子の損失が最も大きく, 酵素の有無によらず 定量PCRでの阻割はほとんど見られなかった. また, リパーゼによる処理を行うことで, 冬期, 春期に採取したカキ 試料によらず, 安定した高効率の回収率が得られた. 以上よりリパーゼを用いたEVE法は, カキ中腸腺中のウイルス 量の季節変動を評価する手法として非常に有用であるといえる.

  104. New tools for the study and direct surveillance of viral pathogens in water 査読有り

    Albert Bosch, Susana Guix, Daisuke Sano, Rosa M. Pinto

    CURRENT OPINION IN BIOTECHNOLOGY　19　(3)　295-301　2008年6月

    出版者・発行元：CURRENT BIOLOGY LTD

    DOI： 10.1016/j.copbio.2008.04.006 　

    ISSN：0958-1669

    詳細を見る 詳細を閉じる

    Half a century ago scientists attempted the detection of poliovirus in water. Since then other enteric viruses responsible for gastroenteritis and hepatitis have replaced enteroviruses as the main target for detection. However, most viral outbreaks are restricted to norovirus and hepatitis A virus, making them the main targets in water.The inclusion of virus analysis in regulatory standards for viruses in water samples must overcome several shortcomings such as the technical difficulties and high costs of virus monitoring, the lack of harmonised and standardised assays and the challenge posed by the ever-changing nature of viruses. However, new tools are nowadays available for the study and direct surveillance of viral pathogens in water that may contribute to fulfil these requirements.

  105. 活性汚泥細菌から分離されたノロウイルス吸着タンパク質 (Norovirus-Binding Proteins: NoVBPs) の特性評価 査読有り

    和田圭史, 佐野大輔, 今井崇博, 大村達夫

    水環境学会誌　30　(12)　731-736　2007年12月

    出版者・発行元：日本水環境学会

    DOI： 10.2965/jswe.30.731 　

    ISSN：0916-8958

    詳細を見る 詳細を閉じる

    In this study, norovirus-binding proteins (NoVBPs) were newly recovered from activated sludge culture using affinity chromatography. The C-terminal amino acid sequence (12 residues) of the Lordsdale virus, belonging to the dominant genotype (GII.4) of noroviruses throughout the world, was used as a ligand in the affinity chromatography. Two-dimensional electrophoresis analysis shows that the samples recovered by the affinity chromatography include a variety of proteins, which have a molecular weights between 60 and 10 kDa and isoelectric points between 3 and 6. The binding affinity of NoVBPs to the virus-like particle (VLP) of norovirus GII.4 was higher than that to the VLP of norovirus GI.1, which was indicated by ELISA. A determination of the N-terminal amino acid sequences revealed that several NoVBPs are relatives to a chaperonine protein, GroEL, which can interact with denatured hydrophobic proteins. The NoVBPs recovered in this study might be useful as versatile adsorbents for norovirus recovery or detection from environmental water samples.

  106. Persistence of caliciviruses in artificially contaminated oysters during depuration 査読有り

    You Ueki, Mika Shoji, Atsushi Suto, Toru Tanabe, Yoko Okimura, Yoshihiko Kikuchi, Noriyuki Saito, Daisuke Sano, Tatsuo Omura

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　73　(17)　5698-5701　2007年9月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.00290-07 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    The fate of calicivirus in oysters in a 10-day depuration was assessed. The norovirus gene was persistently detected from artificially contaminated oysters during the depuration, whereas feline calicivirus in oysters was promptly eliminated. The prolonged observation of norovirus in oysters implies the existence of a selective retention mechanism for norovirus within oysters.

  107. Human sapovirus in clams, Japan 査読有り

    Grant S. Hansman, Tomoichiro Oka, Reiko Okamoto, Tomoko Nishida, Shoichi Toda, Mamoru Noda, Daisuke Sano, You Ueki, Takahiro Imai, Tatsuo Omura, Osamu Nishio, Hirokazu Kimura, Naokazu Takeda

    EMERGING INFECTIOUS DISEASES　13　(4)　620-622　2007年4月

    出版者・発行元：CENTER DISEASE CONTROL

    DOI： 10.3201/eid1304.061390 　

    ISSN：1080-6040

    詳細を見る 詳細を閉じる

    Human sapovirus was detected in 4 of 57 clam packages by reverse transcription-PCR and sequence analysis. This represents the first finding of sapovirus contamination in food. Closely matching sequences have been detected in stool specimens from patients with gastroenteritis in Japan, which indicates a possible food-to-human transmission link.

  108. Cellular proteins of Microcystis aeruginosa inhibiting coagulation with polyaluminum chloride 査読有り

    Tomoko Takaara, Daisuke Sano, Hiroshi Konno, Tatsuo Omura

    Water Research　41　1653-1658.　2007年

    DOI： 10.1016/j.watres.2007.01.035 　

  109. Sapovirus in water, Japan 査読有り

    Grant S. Hansman, Daisuke Sano, You Ueki, Takahiro Imai, Tomoichiro Oka, Kazuhiko Katayama, Naokazu Takeda, Tatsuo Omura

    Emerging Infectious Diseases　13　(1)　133-135　2007年1月

    DOI： 10.3201/eid1301.061047 　

    ISSN：1080-6040

    eISSN：1080-6059

    詳細を見る 詳細を閉じる

    Sapoviruses are etiologic agents of human gastroenteritis. We detected sapovirus in untreated wastewater, treated wastewater, and a river in Japan. A total of 7 of 69 water samples were positive by reverse transcription - PCR. Phylogenetic analysis of the viral capsid gene grouped these strains into 4 genetic clusters.

  110. Genetic variation in the conservative gene region of norovirus genogroup II strains in environmental and stool samples 査読有り

    Daisuke Sano, You Ueki, Toru Watanabe, Tatsuo Omura

    ENVIRONMENTAL SCIENCE & TECHNOLOGY　40　(23)　7423-7427　2006年12月

    出版者・発行元：AMER CHEMICAL SOC

    DOI： 10.1021/es0603453 　

    ISSN：0013-936X

    詳細を見る 詳細を閉じる

    Noroviruses (NoVs) have been one of leading etiological agents for infectious gastroenteritis over the world. Gastroenteritis caused by NoVs is prevalent in winter season, and the contamination of the water environment with NoVs in the epidemic cold season is frequently reported. In contrast, the number of gastroenteritis patients and NoVs in the water environment are reduced during the non-epidemic summer season, and the year-round fate of NoVs has remained to be elucidated. In this study, we collected nucleotide sequences of NoV genogroup II (GII) from domestic sewage, sewage sludge, treated wastewater, river water, and stool samples of gastroenteritis patients in geographically close areas. Phylogenetic analysis of the obtained NoV gene revealed that six out of seven isolates from environmental samples and 10 out of 11 isolates from stool samples belong to genotype 3 (NoV GII. 3) or 4 (NoV GII. 4), which have been prevalent throughout the world. Genetic distances between the conservative gene region of NoV GII. 4 variants implied that genetically diverse strains are likely to occur in environmental samples. The evaluation of the evolutionary change of NoV gene obtained from environmental samples would make it possible to elucidate the year-round fate of NoVs.

  111. 凝集阻害を引き起こすMicrocystis aeruginosa由来親水性物質の分離及び特性評価 査読有り

    高荒智子, 佐野大輔, 須藤丈, 今野弘, 大村達夫

    環境工学研究論文集　43　585-591　2006年11月

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.43.585 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    閉鎖性水域における季節的な藻類の大量増殖は浄水処理において様々な障害を引き起こす. 藻類障害の一つである凝集阻害については詳細な発生メカニズムが明らかにされていないため, 抜本的な解決策が存在しないのが現状である. 本研究では, 藻類由来の凝集阻害物質としてMicrocystis aeruginosa由来の親水性物質に着目し, その分画分離及び特性評価を行った. その結果, フェノールを用いた抽出法により水層に溶出される藻類由来有機物 (algogedc organic matter. AOM) は, 非常に強い凝集阻害を引き起こすことを確認した. この凝集阻害物質はエタノール沈殿処理により沈殿させることが可能であり, 分子量は10kDa以上であること, 及び中性付近のpHでは負に帯電していることを示唆する結果を得た.

  112. ファージディスプレイ法を用いた凝集阻害誘因タンパク質をコードする藻類遺伝子の分離 査読有り

    佐野大輔, 高荒智子, 鈴木孝佳, 大村達夫

    環境工学研究論文集　43　127-133　2006年11月

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.43.127 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    富栄養化によって大量増殖した藻類は, 浄水処理において凝集処理効率の低下や浄水中へのアルミニウム残留等の凝集阻害を引き起こしている. 凝集阻害メカニズムとして藻類由来有機物と凝集剤成分との錯体形成や荷電中和反応の阻害に関する報告はあるが, 凝集阻害誘因物質を同定して凝集阻害メカニズムを解明する試みは少ない. 本研究では藻類由来タンパク質を提示させたファージディスプレイライブラリからポリ塩化アルミニウムに親和性を有するファージを選択し, 得られたファージの保持するDNA配列を解読することで凝集阻害誘因タンパク質の同定を試みた. その結果, セリンやスレオニンなどヒドロキシル基を有するアミノ酸を保持するタンパク質や金属酵素が凝集阻害に関与していることが示唆された.

  113. Cloning of a heavy-metal-binding protein derived from activated-sludge microorganisms 査読有り

    Daisuke Sano, Ken Myojo, Tatsuo Omura

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　72　(9)　6377-6380　2006年9月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.00656-06 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    A gene of the heavy-metal-binding protein (HMBP) was newly isolated from a genetic DNA library of activated-sludge microorganisms. HMBP was produced by transformed Escherichia coli, and the copper-binding ability of HMBP was confirmed. HMBP derived from activated sludge could be available as heavy metal adsorbents in water and wastewater treatments.

  114. 海域におけるヒト病原ウイルス汚染

    佐野大輔, 植木洋, 大村達夫

    日本海水学会誌　60　(4)　229-237　2006年

    出版者・発行元：日本海水学会

    DOI： 10.11457/swsj1965.60.229 　

    ISSN：0369-4550

    詳細を見る 詳細を閉じる

    Human pathogenic viruses such as Noroviruses and Rotaviruses contaminate water environments, including estuary and coastal areas. These pathogenic viruses in sea area would pose risks for infectious diseases among humans who eat contaminated sea products or play in contaminated bathing areas. It is of primary importance to comprehend the actual situation of the virus contamination of sea areas, which could be regarded as one of considerable reservoirs of human pathogenic viruses. In this article, recent knowledge in regards to the fate of human pathogenic viruses in sea areas is reviewed, and the moving pathway of these viruses estimated by scientific evidences is discussed.

  115. 河川及び海域における病原ウイルス汚染-ノロウイルス調査事例

    佐野大輔, 植木洋

    日本水環境学会誌　29　(3)　130-134　2006年

  116. 水中病原ウイルスによる水環境汚染の実態

    佐野大輔, 植木洋, 渡部徹

    モダンメディア　52　(2)　115-124　2006年

  117. Membrane separation of indigenous noroviruses from sewage sludge and treated wastewater 査読有り

    D. Sano, Y. Ueki, T. Watanabe, T. Omura

    WATER SCIENCE AND TECHNOLOGY　54　(3)　77-82　2006年

    出版者・発行元：I W A PUBLISHING

    DOI： 10.2166/wst.2006.451 　

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    In this study, feasibility of membrane separation for the removal of indigenous noroviruses (NVs) is evaluated. The indigenous NV gene was never detected from ultrafiltration (UF) permeates of sewage sludge and treated wastewater. Indigenous NV gene was also not detected from permeates of sewage sludge and treated wastewater by microfiltration (IVIF) with a pore size of 0.1 mu m (MF0.1). Even though the pore size of IVIF (0.1 mu m) was much larger than the diameter of virus particle (approximately 30-40 nm), more than 4-log(10) reduction value (LRV) at maximum was achieved by membrane separation with MF0.1. NV genes were often detected from permeates of sewage sludge and treated wastewater by IVIF with a pore size of 0.45 mu m (MF0.45), although the maximum log(10) reduction values were more than 3.59 for sewage sludge and more than 2.90 for treated wastewater. It is important to verify factors determining the removal efficiency of viruses With MF membranes.

  118. Heavy metal-binding proteins from metal-stimulated bacteria as a novel adsorbent for metal removal technology 査読有り

    D Sano, K Myojo, T Omura

    WATER SCIENCE AND TECHNOLOGY　53　(6)　221-226　2006年

    出版者・発行元：I W A PUBLISHING

    DOI： 10.2166/wst.2006.200 　

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Water pollution with toxic heavy metals is of growing concern because heavy metals could bring about serious problems for not only ecosystems in the water environment but also human health. Some metal removal technologies have been in practical use, but much energy and troublesome treatments for chemical wastes are required to operate these conventional technologies. In this study, heavy metal-binding proteins (HMBPs) were obtained from metal-stimulated activated sludge culture with affinity chromatography using copper ion as a ligand. Two-dimensional electrophoresis revealed that a number of proteins in activated sludge culture were recovered as HMBPs for copper ion. N-termini of five HMBPs were determined, and two of them were found to be newly discovered proteins for which no amino acid sequences in protein databases were retrieved at more than 80% identities. Metal-coordinating amino acids occupied 38% of residues in one of the N-terminal sequences of the newly discovered HMBPs. Since these HMBPs were expected to be stable under conditions of water and wastewater treatments, it would be possible to utilize HMBPs as novel adsorbents for heavy metal removal if mass volume of HMBPs can be obtained with protein cloning techniques.

  119. 活性汚泥細菌ゲノムDNAライブラリから取得した新規重金属吸着タンパク質遺伝子 査読有り

    佐野大輔, 明星賢, 大村達夫

    環境工学研究論文集　42　601-608　2005年12月

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.42.601 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    Gene of heavy metal-binding protein (HIVIBP) was newly isolated from genomic DNA library of activated sludge bacteria. In order to obtain the objective gene on the strength of N-terminal amino acid sequence of HMBP, a set of gene screening method was constructed. The constructed screening method employs CODEHOP method, touchdown PCR and semi-nested PCR, which makes it possible to amplify the specific HMBP gene. Two acidic amino acids (aspartic acid and glutamic acid) occupied 24% of a deduced amino acid sequence of HMBP, and the rate of metal-coordinating amino acids (aspartic acid, glutamic acid, serine, methionine and histidine) among the deduced sequence of HMBP reached 35%. The estimated secondary and tertiary structures revealed that HMIBP is composed of abundance of alpha helices in which amino acid side chains expose to outside. A lot of metal-coordinating residues projecting out from the alpha helices would confer the heavy metal-binding ability on HMBP.

  120. Norovirus pathway in water environment estimated by genetic analysis of strains from patients of gastroenteritis, sewage, treated wastewater, river water and oysters 査読有り

    Y Ueki, D Sano, T Watanabe, K Akiyama, T Omura

    WATER RESEARCH　39　(18)　4271-4280　2005年11月

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1016/j.watres.2005.06.035 　

    ISSN：0043-1354

    詳細を見る 詳細を閉じる

    In this study, Norovirus (NV) capsid gene was detected from patients of gastroenteritis, domestic sewage, treated wastewater, river water and cultivated oysters in geographically close areas where all of samples were collected. In order to improve recovery efficiency of NVs from oysters, a new method using a spallation apparatus was developed. As a result, 18 of 30 oysters (60%) were positive for NV gene, while 7 of 30 (23%) oysters from the same sampling point were positive with the conventional ultracentrifugal method between November 2003 and February 2004. These results indicate that our new method exhibits the higher efficiency of recovering NVs than the conventional ultracentrifugal method. Six of 8 samples (75%) of river water were positive for NV gene between November 2003 and February 2004. Further-more, 8 of 9 samples (89%) of treated wastewater and all 9 samples of sewage were positive for NV gene in the same period. These results indicated that treated wastewater would be one of the main sources for NV pollution in this area. The phylogenctic analysis in isolated NV capsid genes was conducted, in which high identities of gene sequences between NVs from patients, domestic sewage, river water and cultivated oysters were observed. These results implied that there would be a geographically associated circulation of NVs between human and cultivated oysters via water environment. It would be important to quantitatively analyze the moving pathway of NVs, which directly link to the development of a new scheme for preventing water environment and cultivated oysters from NV contamination. (c) 2005 Elsevier Ltd. All rights reserved.

  121. Construction of a cloning system for the mass production of a virus-binding protein specific for poliovirus type 1 査読有り

    D Sano, T Omura

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　71　(5)　2608-2615　2005年5月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.71.5.2608-2615.2005 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    In our previous study, virus-binding proteins (VBPs) demonstrating the ability to strongly bind poliovirus type 1 (PV1) were recovered from a bacterial culture derived from activated sludge. The isolated VBPs would be useful as viral adsorbents for water and wastewater treatments. The VBP gene of activated sludge bacteria was isolated, and the cloning system of the VBP was established. The isolation of the VBP gene from DNA libraries for activated sludge bacteria was achieved with the colony hybridization technique. The sequence of the VBP gene consisted of 807 nucleotides encoding 268 amino acids. Fifteen amino acid sequences were retrieved from 2,137,877 sequences by a homology search using the BLAST server at the National Center for Biotechnology Information. The protein encoded in the isolated genome was considered to be a newly discovered protein from activated sludge culture, because any sequences in protein databases were not perfectly matched with the sequence of the VBP. It was confirmed that Escherichia coli BL21 transformed by pRSET carrying the isolated VBP gene could extensively produce the VBP clones. Enzyme-linked immunosorbent assay (ELISA) revealed that the VBP clone exhibited the binding ability with intact particles of PV1. The equilibrium binding constant between PV1 and VBP in the ELISA well was estimated to be 2.1 x 10(7) (M-1), which also indicated that the VBP clones have a high affinity with the PV1 particle. The VBP cloning system developed in this study would make it possible to produce a mass volume of VBPs and to utilize them as a new material of the specific adsorbent in several technologies, including virus removal, concentration, and detection.

  122. Affinity Isolation of Algal Organic Matters Able to Form Complex with Aluminum Coagulant 査読有り

    Tomoko Takaara, Daisuke Sano, Hiroshi Konno, Tatsuo Omura

    Water Science and Technology: Water Supply　4　(5-6)　95-102　2005年

  123. アデノウイルス吸着タンパク質(Adenovirus‐Binding Protein:ADVBP)の活性汚泥細菌からの分離 査読有り

    佐野大輔, 石井哲平, 大村達夫

    環境工学研究論文集　41　331-337　2004年11月

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.41.331 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    Outbreaks of waterborne infectious diseases caused by pathogenic viruses have been frequently documented over the world. Pathogenic viruses can survive for a long period in water environment, but there has been a difficulty in removing or inactivating viruses in conventional water and wastewater treatment systems. A new technology for the virus removal from water needs to be developed. In our previous study, virus-binding proteins (VBPs) demonstrating the ability to strongly bind Poliovirus type 1 (PV1) were recovered from a bacterial culture derived from activated sludge. It was expected that the isolated VBPs would be useful as viral adsorbents under the conditions of water and wastewater treatments. In order to develop new technology for virus removal, it is important to isolate not only VBPs for PV1 but also VBPs for other important pathogenic viruses in water. In this study, Adenovirus-Binding Proteins (ADVBPs) for Adenoviruses type 3 (AD3) and type 40/41 (AD40/41) were newly isolated from activated sludge culture with the affinity chromatography technique. Three distinct peaks were sequentially obtained in each affinity chromatographic profile of ADVBPs for AD3 and AD40/41, respectively. The isolated ADVBP in the first peak include a number of proteins, and their molecular weights were widely distributed. ELISA revealed that the ability of binding Adenoviral particle of ADVBP in the second and third peaks were stronger than those in the first peak in the affinit chromatographic profile. It is speculated that the isolated ADVBPs can be useful as specific adsorbents for Adenoviruses in several fields including virus removal, concentration and detection.

  124. Virus-binding proteins recovered from bacterial culture derived from activated sludge by affinity chromatography assay using a viral capsid peptide 査読有り

    D Sano, T Matsuo, T Omura

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　70　(6)　3434-3442　2004年6月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.70.6.3434-3442.2004 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    The contamination of water environments by pathogenic viruses has raised concerns about outbreaks of viral infectious diseases in our society. Because conventional water and wastewater treatment systems are not effective enough to inactivate or remove pathogenic viruses, a new technology for virus removal needs to be developed. In this study, the virus-binding proteins (VBPs) in a bacterial culture derived from activated sludge were successfully recovered. The recovery of VBPs was achieved by applying extracted crude proteins from a bacterial culture to an affinity column in which a custom-made peptide of capsid protein from the poliovirus type 1 (PV1) Mahoney strain (H2N-DNPASTTNKDKL-COOH) was immobilized as a ligand. VBPs exhibited the ability to adsorb infectious particles of PV1 Sabin I as determined by enzyme-linked immunosorbent assay. The evaluation of surface charges of VBPs with ion-exchange chromatography found that a majority of VBP molecules had a net negative charge under the conditions of affinity chromatography. On the other hand, a calculated isoelectric point implied that the viral peptide in the affinity column was also charged negatively. As a result, the adsorption of the VBPs to the viral peptide in the affinity column occurred with a strong attractive force that was able to overcome the electrostatic repulsive force. Two-dimensional electrophoresis revealed that the isolated VBPs include a number of proteins, and their molecular masses were widely distributed but smaller than 100 kDa. Amino acid sequences of N termini of five VBPs were determined. Homology searches for the N termini against all protein sequences in the National Center for Biotechnology Information (NCBI) database showed that the isolated VBPs in this study were newly discovered proteins. These VBPs that originated with bacteria in activated sludge might be stable, because they are existing in the environment of wastewater treatments. Therefore, a virus removal technology utilizing VBPs as viral adsorbents can be developed, since it is possible to replicate VBPs by protein cloning techniques.

  125. 凝集阻害を誘因する藻類由来タンパク質の同定を目的としたファージディスプレイの構築 査読有り

    鈴木孝佳, 高荒 智子, 佐野大輔, 大村達夫

    環境工学研究論文集　41　339-346　2004年

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.41.339 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    Seasonal overgrowth of cyanobacteria such as <I>Microcystis aeruginosa (M. aeruginosa</I>) is one of serious problems in semi-closed water area including reservoirs. It is known that algogenic organic matters (AOMs) bring about the inhibition of coagulation on the flocculation process in drinking water treatment systems. Our previous studies have shown that some kinds of protein in AOMs are responsible for the inhibition of coagulation, although the mechanisms of the inhibitoiy effects of algal proteins remain unclear. In order to reveal the inhibitory mechanism of algal proteins, it is necessary to identify the algal proteins closely related to the inhibition of coagulation. In this study, phage display using the genome of <I>M. aeruginosa</I> was constructed to extract algal proteins that exhibit the inhibitory effect on the coagulation. <I>M. eruginosa</I> cells on steady growth phase were collected, and genomic DNA of M. aeruginosa cells was extracted. Thean, extracted algal DNA was digested by the restriction enzyme Sau3AI. The phagemid vecter named pSKAN8-EWQ was newly created, in which BamHI recognition site was included as a single site for the restriction enzyme. Digested genomic DNA of<I> M. aeruginosa</I> was ligated with pSKAN8-EWQ, and the ligated phagemid vector was used to transform the competent cells of E. coli JM109. The cocktail of phages displaying algal proteins was obtained by introducing the helperphage (VCSM13 Interference-Resistant Helper Phage) into the transformed E. coli cells. It was confirmed that obtained phages are able to infect E. coli. JM109, and transform E. coli. cells into the ampicilin-resistant strain. This thing indicated that the phage display system for algal proteins was successfully constructed. The phage display established in this study will definitely contribute to identifying the inhibitory algal proteins on the coagulation, and revealing the inhibitoiy mechanism in the further study.

  126. Detection of infectious pathogenic viruses in water and wastewater samples from urbanised areas 査読有り

    D Sano, T Watanabe, T Matsuo, T Omura

    WATER SCIENCE AND TECHNOLOGY　50　(1)　247-251　2004年

    出版者・発行元：I W A PUBLISHING

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Regardless of the remarkable improvement in sanitary conditions in urbanised areas, infectious diseases caused by pathogenic viruses in water have been reported year after year. The actual situation for the behaviour of pathogenic viruses in urbanised society should be understood. In this study, infectious enteroviruses and adenoviruses in water and wastewater samples from urbanised areas were investigated. Infectious enteroviruses were continuously detected from sewage and sewage sludge during the summer season. The detection of infectious viruses in sewage and sewage sludge was facilitated by the enzymatic virus elution (EVE) method. The concentration of infectious enteroviruses in positive samples of sewage and sewage sludge was &gt;400 virions/L. Infectious viruses were also detected from water samples at levels &gt;4 virions/L.

  127. アルミニウム錯体形成能を有する藻類由来有機物の分離 査読有り

    高荒智子, 丸山亜紀子, 佐野大輔, 今野弘, 大村達夫

    環境工学研究論文集　40　227-236　2003年11月

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.40.227 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    Mass propagation of algae in raw water has been known related to disturbing the flocculation in drinking watertreatmentsystems. Many researchers proposed that algal organic matters (AOMs) can interact with flocculants by coordinate bonds, and bring about several disorders due to increases in flocculant demand and in the residual flocculant in treated water. However, the inhibitory mechanism by AOMs have not been elucidated In this study, alminium-complexing substances in AOMs were successfully isolated with affinity chromatography. SDS-PAGE analysis revealed that these substances include proteins which have molecular weight between 40 and 70kDa. However, the results of the gel chromatography show that the amount of these proteins in AOMs were estimated to be very low compared to organic matters with high molecular weights around 2000kDa. These results imply that not only proteins but also other organic matters such as polysaccharides contribute to the inhibition of flocculation.

  128. Detection of enteric viruses in municipal sewage sludge by a combination of the enzymatic virus elution method and RT-PCR 査読有り

    D Sano, K Fukushi, Y Yoshida, T Omura

    WATER RESEARCH　37　(14)　3490-3498　2003年8月

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1013/S0043-1354(03)00208-2 　

    ISSN：0043-1354

    詳細を見る 詳細を閉じる

    Pathogenic enteric viruses can be retained in municipal sewage sludge as has been reported by many researchers. Although the RT-PCR technique has been extensively employed for the virus detection from various environmental samples, the application of RT-PCR to the detection of viruses in sewage sludge has the difficulty because of inhibitory substances to the gene amplification. However, a combination of the enzymatic virus elution (EVE) method with RTPCR made it possible to effectively detect viruses in sewage sludge. The enzymatic breakdown of sludge flocs in the EVE method enhanced the virus elution from poliovirus 1 (PV1)-inoculated sewage sludge, and the detection of PV1 was performed by RT-PCR without any inhibitions. On the contrary, the application of RT-PCR to the viral assay in the USEPA method using the 10% beef extract solution was not practical because of inhibitions to the viral gene amplification. The combination of the EVE method using lysozyme (polysaccharide-degrading enzyme), papain (protease), and chymotrypsin (protease) with RT-PCR resulted in a virus recovery efficiency of 31%, but a synergistic effect of these enzymes on the virus recovery efficiency was not observed. The EVE method using lysozyme or papain could be a promising procedure for the virus elution from sewage sludge in detecting these viruses with RT-PCR. (C) 2003 Elsevier Science Ltd. All rights reserved.

  129. Functional metal-binding proteins by metal-stimulated bacteria for the development of an innovative metal removal technology 査読有り

    T Antsuki, D Sano, T Omura

    WATER SCIENCE AND TECHNOLOGY　47　(10)　109-115　2003年

    出版者・発行元：I W A PUBLISHING

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Heavy metal pollution has become an environmental problem throughout the world because heavy metals can be accumulated into the food chain and bring about serious problems, not only for ecosystems but also for human health. In this study, functional metal-binding proteins (FMBPs) were isolated from a metal-stimulated activated sludge culture with the aim of applying them to an innovative metal removal technology. Activated sludge bacteria was cultured in growth media including copper ion, and the stimulation of protein production by copper ion led to the 14% increase in a quantity of extracted crude proteins per 1 g of bacterial cell pellet (wet). In order to isolate FMBPs,. extracted crude proteins were. applied to the immobilized metal affinity column in which each of copper, nickel and zinc was used as a ligand. Several FMBPs were succesfully isolated from copper-stimulated bacteria. One of FMBPs. (molecular weight of about 40 kDa) exhibited an ability to adsorb all three metals. The multi metal-binding property of this FMBP could be applied to an innovative metal removal technology: Furthermore, isolated. FMBPs that could capture only one kind of heavy metal would also be attractive as a metal adsorbent in recovering a specific metal as a resource from wastewater, including several heavy metals.

  130. アフィニティクロマトグラフィによる活性汚泥からのウイルス吸着タンパク質（Virus-Binding Protein：VBP）の分離と特性評価 査読有り

    松尾崇宏, 佐野大輔, 大村達夫

    環境工学研究論文集　39　345-353　2002年

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.39.345 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    Water pollution with pathogenic viruses has been frequently reported in recent years. Since conventionalwater and wastewater treament systems are not enough to inactivate or remove pathogenic viruses, a new technology for virus removal should be developed. In this study, virus-binding proteins (VBPs) were isolated from activated sludge culture with affinity chomatography. The ability of VBPs to capture intact particles of poliovirus I was confirmed with ELISA. SDS-PAGE analysis revealed that VBPs included a number of polypeptidesthat have molecular weight between 94.0 and 14.4k Da. The evaluation of surface charges of VBPs with ion exchange chromatography found that a majority of VBP molecules had a net negative charge under the condition of the affinity chromatography. On the other hand, the viral peptide, which was used as an affinity ligand, was expected to have a net negative charge. As a result, the VBPs must be captured by the viral peptide with stronger binding forces than the electrostatic repulsive force. The affinity adsorption could explain the strong attractive interaction between VBPs and viral peptide. These VBPs could be useful as an innovative material for the virus removal.

  131. Risk evaluation for pathogenic bacteria and viruses in sewages sludge compost 査読有り

    T. Watanabe, D. Sano, T. Omura

    Water Science and Technology　46　(11-12)　325-330　2002年

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Composting can be regarded as the most available option for recycling of swage sludge. However, the existence of pathogenic bacteria and viruses in the compost has been scarcely investigated until now. So there is little information on the infectious risk through agricultural activities or gardening in using the compost. In this study, several kinds of composts were investigated for detection of pathogenic bacteria (Salmonella spp. and Escherichia coli O157) and enteric viruses. It was concluded from the result that these bacteria and viruses could not be detected in 1.0 g-wet of any kinds of composts. Infectious risks through agricultural activities or gardening were evaluated by Monte Carlo simulation in the case that the compost was polluted by Samonella spp., E. coli O157:117 and Poliovirus 1. Criteria satisfying the acceptable risk (less than 10-4 per year) for these pathogenic bacteria and virus in the compost were determined from the result of simulations. 1.0 [CFU or PFU/g-wet] was available as the criteria for E. coli O157 and Poliovirus 1 in the compost. On the other hand, the criterion for Salmonella spp. in the compost should be established on a lower concentration than 0.001 CFU/g-wet.

  132. Enhanced virus recovery front municipal sewage sludge with a combination of enzyme and cation exchange resin 査読有り

    D Sano, K Fukushi, K Yano, Y Yoshida, T Omura

    WATER SCIENCE AND TECHNOLOGY　43　(2)　75-82　2001年

    出版者・発行元：I W A PUBLISHING

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    There is a great difficulty in virus enumeration in sewage sludge because viruses in sludge are firmly captured by sludge solids. In order to determine the precise number of viruses in sludge, an enhanced virus recovery method with a combination of an enzyme and a cation exchange resin (CER) was developed. Test viruses were seeded to a sample sludge obtained from a municipal wastewater treatment plant, and the sludge were incubated with various eluents. The quantity of eluted viruses in the liquid phase was then measured by the plaque assay technique. Using the eluent containing only water, CER, and CER with enzyme exhibited 0 %, 19 % and 39 % of virus recovery, respectively. While the conventional USEPA method exhibited a virus recovery of 21 %. Furthermore, viruses eluted by the eluent containing the CER and the lysozyme included not only surface-attached viruses but also solids-embedded viruses.

  133. 銅吸着能を有する活性汚泥中タンパク質の検索 査読有り

    安附太郎, 佐野大輔, 福士謙介, 大村達夫

    環境工学研究論文集　37　131-140　2000年11月

    出版者・発行元：Japan Society of Civil Engineers

    DOI： 10.11532/proes1992.37.131 　

    ISSN：1341-5115

    詳細を見る 詳細を閉じる

    The toxic metal pollution is one of the major problems in environmental waters. Although the metal pollution has not been recently reported because of the development of successful removal technologies, the problem at relatively low concentration of metals still remains. The enhanced recovery of metals from water and wastewater becomes the social needs because metals have the toxicity to humans even at such low concentrations. Metals in wastewater adsorb easily to proteins in activated sludge and metals adsorbed are immediately dissociated with the increase of hydrogen ion concentration. These properties are available for both a metal recovery from environmental waters and a recycling after the metal recovery.<BR>In this study, proteins that had the high affinity to copper (copper-binding proteins) were isolated from activated sludge culture using the affinity chromatography. As the result of the SDS-PAGE, the proteins were divided into two groups by their molecular weights. One group was from 43kDa to 30kDa and another was less than 30kDa. The latter group was the copper-binding proteins produced when copper ion was present as an inducer in the activated sludge culture.

︎全件表示 ︎最初の5件までを表示

MISC 64

  1. 総説・水環境学の進歩-研究委員会編(11)水中の健康関連微生物研究委員会の研究・活動紹介

    片山浩之, 原本英司, 佐野大輔, 渡部徹, 西山正晃, 井原賢, 端昭彦, 五味良太, 鳥居将太郎, 安井碧, 大石若菜

    水環境学会誌　46(A)　(12)　2023年

    ISSN：0916-8958

  2. ウイルス感染症とヒトへの感染経路と感染対策に関する考察-空気を介した感染を防ぐために-

    高塚威, 宇田川洋一, 北島正章, 佐野大輔, 宇田川悦子

    空気調和・衛生工学会論文集　(286)　2021年

    ISSN：0385-275X

  3. 集団内の遺伝的多様性がもたらすロタウイルス株の遊離塩素感受性変化

    門屋俊祐, 西村修, 佐野大輔, 浦山俊一, 布浦拓郎, 北島正章, 岡部聡, 中込とよ子, 中込治

    日本水環境学会年会講演集　54th　2020年

  4. 水系感染症ウイルスの集団内遺伝的多様性がもたらす塩素感受性変化

    門屋俊祐, 浦山俊一, 布浦拓郎, 中込治, 中込とよ子, 北島正章, 岡部聡, 西村修, 佐野大輔

    日本水環境学会シンポジウム講演集　23rd　2020年

  5. ウイルス感染症とヒトへの感染と空気-空気を介した感染を防ぐために-

    高塚威, 宇田川洋一, 北島正章, 佐野大輔, 宇田川悦子, 宇田川悦子

    新日本空調技術開発研究所技報　(25)　2020年

  6. 共存細菌との結合が水系感染症ウイルスの感染効率に与える影響

    河合大樹, 北島正章, 岡部聡, 佐野大輔

    日本水環境学会年会講演集　52nd　2018年

  7. DNAアプタマーを用いた簡易ノロウイルス検出法の開発

    吉原光, 北島正章, 岡部聡, 高橋正宏, 佐藤久, 佐野大輔

    日本水環境学会年会講演集　52nd　2018年

  8. 環境水から単離した溶菌性バクテリオファージによる水中病原細菌の低減効果

    高木達馬, 佐野大輔, 岡部聡, 北島正章

    日本水環境学会年会講演集　52nd　2018年

  9. ウイルス干渉現象がIntegrated cell culture-PCRによる感染性ウイルス検出に与える影響

    渡邊亮介, 北島正章, 岡部聡, 佐野大輔

    土木学会北海道支部論文報告集(CD-ROM)　(74)　2018年

  10. ウイルス干渉現象がIntegrated cell culture-PCRによる感染性アデノウイルス検出に与える影響

    佐野大輔, 渡邊亮介, 北島正章, 岡部聡

    日本水環境学会シンポジウム講演集　21st　2018年

  11. ナノ流路デバイスを用いた下水中RNAウイルス定量およびタイピング:DHSリアクターへの適用例

    押木守, 小林直央, 荒木信夫, 伊藤寿宏, 佐野大輔, 三浦尚之, 風間しのぶ, 原田秀樹, 久保田健吾, 幡本将史, 山口隆司, 井口晃徳, 瀬川高弘, 加藤毅, 多川正, 大久保努, 上村繁樹, 高橋優信

    日本水環境学会年会講演集　51st　546　2017年3月9日

  12. 遊離塩素耐性ロタウイルスの変異遺伝子同定に関する研究

    門屋俊祐, 浦山俊一, 布浦拓郎, 北島正章, 岡部聡, 佐野大輔

    日本水環境学会年会講演集　51st　2017年

  13. DNAアプタセンサー:水中ノロウイルスの即時・高感度検出に適した新技術

    平野誠也, 岡本章玄, 佐野大輔, 岡部聡, 北島正章

    日本水環境学会年会講演集　51st　2017年

  14. 病原性レジオネラの水系感染制御に資する溶菌性バクテリオファージの探索

    高木達馬, 佐野大輔, 岡部聡, 北島正章

    日本水環境学会年会講演集　51st　2017年

  15. 水環境におけるノロウイルスの遺伝子型別存在実態の把握

    宮村明帆, 植木洋, 三浦尚之, 風間しのぶ, 北島正章, 岡部聡, 佐野大輔

    日本水環境学会年会講演集　51st　2017年

  16. ウイルス吸着性細菌との結合が水系感染症ウイルスの感染能力に与える影響

    河合大樹, 中込とよ子, 中込治, 北島正章, 岡部聡, 佐野大輔

    日本水環境学会年会講演集　51st　2017年

  17. 環境水中からのノロウイルス吸着性大腸菌の単離と吸着因子同定に関する研究

    羽柴聡, AMARASIRI Mohan, 原本英司, 北島正章, 岡部聡, 佐野大輔

    日本水環境学会年会講演集　51st　2017年

  18. 遊離塩素耐性ロタウイルスの変異遺伝子および耐性メカニズムの解明

    門屋俊祐, 浦山俊一, 布浦拓郎, 北島正章, 岡部聡, 佐野大輔

    環境工学研究フォーラム講演集　54th　2017年

  19. 水系感染性病原細菌に対するバクテリオファージの単離および溶菌特性解析

    高木達馬, 佐野大輔, 岡部聡, 北島正章

    環境工学研究フォーラム講演集　54th　2017年

  20. DNAアプタマーを用いた簡易ノロウイルス検出法の開発

    吉原光, 北島正章, 佐野大輔, 岡部聡, 高橋正宏, 佐藤久

    環境工学研究フォーラム講演集　54th　2017年

  21. 下水再生利用のためのノロウイルス目標除去効率算定に関する研究

    伊藤寿宏, 北島正章, 加藤毅, 石井聡, 瀬川高弘, 岡部聡, 佐野大輔

    日本微生物生態学会大会(Web)　2017　2017年

  22. 組織細胞におけるイオンチャネル遺伝子発現プロファイルに着目した水中感染性エンテロウイルス迅速検出手法の開発

    渡邊亮介, 稲葉愛美, 岡部聡, 佐野大輔, 北島正章

    日本水環境学会年会講演集　51st　2017年

  23. DNAアプタマーを用いた簡易ノロウイルス検出法の開発

    吉原光, 北島正章, 佐野大輔, 岡部聡, 高橋正宏, 佐藤久

    全国会議(水道研究発表会)講演集　2017　2017年

    ISSN：2436-1496

  24. Aptamer: The potential application to norovirus research, diagnosis, and therapeutics

    Seiya Hirano, Daisuke Sano, Satoshi Okabe, Masaaki Kitajima

    Noroviruses: Outbreaks, Control and Prevention Strategies　223-242　2017年1月1日

    詳細を見る 詳細を閉じる

    © 2017 Nova Science Publishers, Inc. Aptamer has been attracting much attention as a new biomolecular tool for viral diagnostic and therapeutic applications. Aptamer is a functional nucleic acid that folds into a unique three-dimensional conformation that binds to a specific target, which can be an alternative for antibodies. Rather, aptamer offers various advantages over antibodies as a target-specific recognition molecule: high affinity, specificity, and thermostability, low production cost, etc. Several studies have reported DNA aptamers for certain human norovirus strains as well as murine norovirus, which has enabled development and evaluation of aptasensor (aptamer-based biosensor) technology for rapid and sensitive norovirus detection. Therapeutic application of aptamer to inhibiting norovirus infections has not been clearly demonstrated to date, but there have been a few reports suggesting the potential of aptamer-based therapeutics. Taken together, it has been implied that aptamer is a new target-specific biomolecular tool that offers various potential applications to norovirus research, diagnosis, and therapeutics, which should contribute to the infection and disease control of norovirus by rapidly identifying potential infection routes and reducing the disease burden through therapeutic approach. Herein we summarize recent development and application of norovirus-specific aptamers and discuss future research directions towards better control of norovirus transmission/infections using this aptamer technology.

  25. DHS(downflow hanging sponge)リアクターにおける病原ウイルスの除去効率

    小林直央, 押木守, 荒木信夫, 本山毅宜, 伊藤寿宏, 佐野大輔, 瀬川高弘, 幡本将史, 山口隆司, 井口晃徳, 上村繁樹, 大久保努, 多川正, 高橋優信, 久保田健吾, 原田秀樹

    日本水環境学会年会講演集　50th　657　2016年3月10日

  26. 遊離塩素処理がノロウイルスの遺伝的多様性に与える影響

    中村新, 渡辺幸三, 八重樫咲子, 岡部聡, 中込とよ子, 中込治, 佐野大輔

    日本水環境学会年会講演集　50th　398　2016年3月10日

  27. ノロウイルスの新知見

    門屋俊祐, 伊藤寿宏, 佐野大輔

    膜協会ジャーナル　23　(1)　7-15　2016年

  28. ノロウイルスの塩素消毒耐性獲得メカニズム解明に関する基礎的研究

    富岡哲史, 渡辺幸三, 岡部聡, 佐野大輔

    土木学会北海道支部論文報告集(CD-ROM)　(70)　ROMBUNNO.G-01　2014年2月

  29. 表面プラズモン共鳴を利用した病原微生物バイオセンサの開発

    坂槙有紀恵, 山田健太, 佐野大輔, 高橋正宏, 岡部聡, 佐藤久

    日本水環境学会年会講演集　47th　436　2013年3月11日

  30. 表面プラズモン共鳴を利用した病原微生物バイオセンサの開発

    坂槙有紀恵, 山田健太, ピタックティーラタム ニティ, 石井聡, 佐野大輔, 高橋正宏, 岡部聡, 佐藤久

    環境工学研究フォーラム講演集　49th　28-30　2012年11月28日

  31. 表面プラズモン共鳴を利用した水中病原微生物検出バイオセンサの開発

    佐藤久, 坂槙有紀恵, 山田健太, ピタックティーラタムニティ, 石井聡, 佐野大輔, 高橋正宏, 岡部聡

    日本水環境学会シンポジウム講演集　15th　233-234　2012年9月10日

  32. 活性汚泥からのウイルス吸着タンパク質分離方法の新規開発

    今井崇博, 佐野大輔, 真砂佳史, 大村達夫

    日本水環境学会年会講演集　45th　441　2011年3月18日

  33. Norovirus-binding proteins recovered from activated sludge micro-organisms with an affinity to a noroviral capsid peptide

    D. Sano, K. Wada, T. Imai, Y. Masago, T. Omura

    JOURNAL OF APPLIED MICROBIOLOGY　109　(6)　1923-1928　2010年12月

    出版者・発行元：WILEY-BLACKWELL PUBLISHING, INC

    DOI： 10.1111/j.1365-2672.2010.04821.x 　

    ISSN：1364-5072

    詳細を見る 詳細を閉じる

    Aims: Transmission routes of noroviruses, leading aetiological agents of acute gastroenteritis, are rarely verified when outbreaks occur. Because the destination of norovirus particles being firmly captured by micro-organisms could be totally different from that of those particles moving freely, micro-organisms with natural affinity ligands such as virus-binding proteins would affect the fate of viruses in environment, if such microbial affinity ligands exist. The aim of this study is to identify norovirus-binding proteins (NoVBPs) that are presumably working as natural ligands for norovirus particles in water environments. Methods and Results: NoVBPs were recovered from activated sludge microorganisms by an affinity chromatography technique in which a capsid peptide of norovirus genogroup II (GII) was immobilized. The recovered NoVBPs bind to norovirus-like particles (NoVLPs) of norovirus GII, and this adsorption was stronger than that to NoVLPs of norovirus genogroup I. The profile of two-dimensional electrophoresis of NoVBPs showed that the recovered NoVBPs included at least seven spots of protein. The determination of N-terminal amino acid sequences of these NoVBPs revealed that hydrophobic interactions could contribute to the adsorption between NoVBPs and norovirus particles. Conclusions: NoVBPs conferring a high affinity to norovirus GII were successfully isolated from activated sludge micro-organisms. Significance and Impact of the Study: NoVBPs could be natural viral ligands and play an important role in the NoV transmission.

  34. Detection of Sapovirus in oysters

    You Ueki, Mika Shoji, Yoko Okimura, Yasuko Miyota, Yoshifumi Masago, Tomoichiro Oka, Kazuhiko Katayama, Naokazu Takeda, Mamoru Noda, Takayuki Miura, Daisuke Sano, Tatsuo Omura

    MICROBIOLOGY AND IMMUNOLOGY　54　(8)　483-486　2010年8月

    出版者・発行元：WILEY-BLACKWELL

    DOI： 10.1111/j.1348-0421.2010.00239.x 　

    ISSN：0385-5600

    詳細を見る 詳細を閉じる

    SaV sequences which are either genetically identical or similar were detected from oysters, feces from gastroenteritis patients, and domestic wastewater samples in geographically close areas. This is the first report of the detection of SaV in oysters which meet the legal requirements for raw consumption in Japan.

  35. Surface-retained organic matter of Microcystis aeruginosa inhibiting coagulation with polyaluminum chloride in drinking water treatment

    Tomoko Takaara, Daisuke Sano, Yoshifumi Masago, Tatsuo Omura

    WATER RESEARCH　44　(13)　3781-3786　2010年7月

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1016/j.watres.2010.04.030 　

    ISSN：0043-1354

    詳細を見る 詳細を閉じる

    Algogenic organic matter produced by the excess growth of cyanobacteria in semi-closed water areas causes coagulation inhibition in drinking water production. In this study, hydrophilic substances of Microcystis aeruginosa, which were mainly composed of lipopolysaccharide (LPS) and RNA, were prepared, and the involvement of these cyanobacterial hydrophilic substances in coagulation inhibition was investigated. As a result, it was found that the negatively charged hydrophilic substances with a molecular weight higher than 10 kDa have a significant role in coagulation inhibition. Further fractionation of cyanobacterial hydrophilic substances revealed that surface-retained organic matter (SOM), including LPS, could exhibit a potent inhibitory effect on the coagulation using polyaluminum chloride (PACl), presumably because of the direct interaction of hydrophilic SOM with cations originated from PACl, which could impede the hydrolysis of the coagulant. (C) 2010 Elsevier Ltd. All rights reserved.

  36. 環境水中におけるノロウイルス(NoV)の挙動

    植木洋, 庄司美加, 上村弘, 沖村容子, 御代田恭子, 今井崇博, 三浦尚之, 真砂佳史, 大村達夫, 佐野大輔, 西尾治

    日本水環境学会シンポジウム講演集　11th　182-183　2008年9月17日

  37. ガラスビーズへのウイルス吸着タンパク質(Virus‐Binding Prote in:VBP)固定化手法の確立

    今井崇博, 真砂佳史, 佐野大輔, 大村達夫

    土木学会年次学術講演会講演概要集(CD-ROM)　63rd　(Disk 2)　ROMBUNNO.7-160　2008年8月13日

  38. New tools for the study and direct surveillance of viral pathogens in water

    Albert Bosch, Susana Guix, Daisuke Sano, Rosa M. Pinto

    CURRENT OPINION IN BIOTECHNOLOGY　19　(3)　295-301　2008年6月

    出版者・発行元：CURRENT BIOLOGY LTD

    DOI： 10.1016/j.copbio.2008.04.006 　

    ISSN：0958-1669

    詳細を見る 詳細を閉じる

    Half a century ago scientists attempted the detection of poliovirus in water. Since then other enteric viruses responsible for gastroenteritis and hepatitis have replaced enteroviruses as the main target for detection. However, most viral outbreaks are restricted to norovirus and hepatitis A virus, making them the main targets in water.The inclusion of virus analysis in regulatory standards for viruses in water samples must overcome several shortcomings such as the technical difficulties and high costs of virus monitoring, the lack of harmonised and standardised assays and the challenge posed by the ever-changing nature of viruses. However, new tools are nowadays available for the study and direct surveillance of viral pathogens in water that may contribute to fulfil these requirements.

  39. Enzymatic Virus Elution(EVE)法による牡蠣中腸腺からのウイルス誘出技術の検討

    今井崇博, 奥村千恵, 佐野大輔, 真砂佳史, 大村達夫

    日本水環境学会年会講演集　42nd　460　2008年3月19日

  40. ウイルスの分析及び膜分離活性汚泥法によるウイルス除去性能評価に関する研究

    大村 達夫, 渡部 徹, 佐野 大輔, 植木 洋

    財団法人建設工学研究振興会年報　42　1-6　2007年4月1日

    出版者・発行元：東北大学

    ISSN：0288-9994

  41. Cellular proteins of Microcystis aeruginosa inhibiting coagulation with polyaluminum chloride

    Tomoko Takaara, Daisuke Sano, Hiroshi Konno, Tatsuo Omura

    WATER RESEARCH　41　(8)　1653-1658　2007年4月

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1016/j.watres.2007.01.035 　

    ISSN：0043-1354

    詳細を見る 詳細を閉じる

    Cyanobacterial growth in semi-closed water areas such as reservoirs brings about a coagulation inhibition in a drinking water treatment system, but the inhibitory substances and mechanisms involved have yet to be elucidated. In this study, proteins having a high affinity with polyaluminum chloride (PACl) were isolated from organic substances produced by Microcystis aeruginosa with the affinity chromatography technique. Both extracellular organic matter (EOM) and cellular organic matter (COM) disturbed the flocculation of suspended kaolin with PACl, but it was likely that nonproteinous substances in EOM cause the reduction of coagulation effciency. In contrast, proteins in COM were obtained as possible inhibitory substances for the coagulation with PACl. These proteins could consume PACl in the coagulation process due to the formation of chelate complexes between these inhibitory proteins and the coagulant. The consumption of PACl by cyanobacterial proteins could be one of the important causes of the increase in coagulant demand. (c) 2007 Elsevier Ltd. All rights reserved.

  42. Human sapovirus in clams, Japan

    Grant S. Hansman, Tomoichiro Oka, Reiko Okamoto, Tomoko Nishida, Shoichi Toda, Mamoru Noda, Daisuke Sano, You Ueki, Takahiro Imai, Tatsuo Omura, Osamu Nishio, Hirokazu Kimura, Naokazu Takeda

    EMERGING INFECTIOUS DISEASES　13　(4)　620-622　2007年4月

    出版者・発行元：CENTER DISEASE CONTROL

    DOI： 10.3201/eid1304.061390 　

    ISSN：1080-6040

    詳細を見る 詳細を閉じる

    Human sapovirus was detected in 4 of 57 clam packages by reverse transcription-PCR and sequence analysis. This represents the first finding of sapovirus contamination in food. Closely matching sequences have been detected in stool specimens from patients with gastroenteritis in Japan, which indicates a possible food-to-human transmission link.

  43. Virus-binding in water environments: natural ligands for human viruses

    D. Sano, T. Omura

    Environmental Microbiology Research Trends　257-272　2007年

  44. Sapovirus in water, Japan

    Grant S. Hansman, Daisuke Sano, You Ueki, Takahiro Imai, Tomoichiro Oka, Kazuhiko Katayama, Naokazu Takeda, Tatsuo Omura

    EMERGING INFECTIOUS DISEASES　13　(1)　133-135　2007年1月

    出版者・発行元：CENTER DISEASE CONTROL

    DOI： 10.3201/eid1301.061047 　

    ISSN：1080-6040

    詳細を見る 詳細を閉じる

    Sapoviruses are etiologic agents of human gastroenteritis. We detected sapovirus in untreated wastewater, treated wastewater, and a river in Japan. A total of 7 of 69 water samples were positive by reverse transcription-PCR. Phylogenetic analysis of the viral capsid gene grouped these strains into 4 genetic clusters.

  45. Genetic variation in the conservative gene region of norovirus genogroup II strains in environmental and stool samples

    Daisuke Sano, You Ueki, Toru Watanabe, Tatsuo Omura

    ENVIRONMENTAL SCIENCE & TECHNOLOGY　40　(23)　7423-7427　2006年12月

    出版者・発行元：AMER CHEMICAL SOC

    DOI： 10.1021/es0603453 　

    ISSN：0013-936X

    詳細を見る 詳細を閉じる

    Noroviruses (NoVs) have been one of leading etiological agents for infectious gastroenteritis over the world. Gastroenteritis caused by NoVs is prevalent in winter season, and the contamination of the water environment with NoVs in the epidemic cold season is frequently reported. In contrast, the number of gastroenteritis patients and NoVs in the water environment are reduced during the non-epidemic summer season, and the year-round fate of NoVs has remained to be elucidated. In this study, we collected nucleotide sequences of NoV genogroup II (GII) from domestic sewage, sewage sludge, treated wastewater, river water, and stool samples of gastroenteritis patients in geographically close areas. Phylogenetic analysis of the obtained NoV gene revealed that six out of seven isolates from environmental samples and 10 out of 11 isolates from stool samples belong to genotype 3 (NoV GII. 3) or 4 (NoV GII. 4), which have been prevalent throughout the world. Genetic distances between the conservative gene region of NoV GII. 4 variants implied that genetically diverse strains are likely to occur in environmental samples. The evaluation of the evolutionary change of NoV gene obtained from environmental samples would make it possible to elucidate the year-round fate of NoVs.

  46. Cloning of a heavy-metal-binding protein derived from activated-sludge microorganisms

    Daisuke Sano, Ken Myojo, Tatsuo Omura

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　72　(9)　6377-6380　2006年9月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.00656-06 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    A gene of the heavy-metal-binding protein (HMBP) was newly isolated from a genetic DNA library of activated-sludge microorganisms. HMBP was produced by transformed Escherichia coli, and the copper-binding ability of HMBP was confirmed. HMBP derived from activated sludge could be available as heavy metal adsorbents in water and wastewater treatments.

  47. 膜処理による下水処理水中のノロウイルス除去に関する研究

    大村 達夫, 渡部 徹, 佐野 大輔, 植木 洋

    財団法人建設工学研究振興会年報　41　28-32　2006年4月1日

    出版者・発行元：東北大学

    ISSN：0288-9994

  48. Heavy metal-binding proteins from metal-stimulated bacteria as a novel adsorbent for metal removal technology

    D Sano, K Myojo, T Omura

    WATER SCIENCE AND TECHNOLOGY　53　(6)　221-226　2006年

    出版者・発行元：I W A PUBLISHING

    DOI： 10.2166/wst.2006.200 　

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Water pollution with toxic heavy metals is of growing concern because heavy metals could bring about serious problems for not only ecosystems in the water environment but also human health. Some metal removal technologies have been in practical use, but much energy and troublesome treatments for chemical wastes are required to operate these conventional technologies. In this study, heavy metal-binding proteins (HMBPs) were obtained from metal-stimulated activated sludge culture with affinity chromatography using copper ion as a ligand. Two-dimensional electrophoresis revealed that a number of proteins in activated sludge culture were recovered as HMBPs for copper ion. N-termini of five HMBPs were determined, and two of them were found to be newly discovered proteins for which no amino acid sequences in protein databases were retrieved at more than 80% identities. Metal-coordinating amino acids occupied 38% of residues in one of the N-terminal sequences of the newly discovered HMBPs. Since these HMBPs were expected to be stable under conditions of water and wastewater treatments, it would be possible to utilize HMBPs as novel adsorbents for heavy metal removal if mass volume of HMBPs can be obtained with protein cloning techniques.

  49. Membrane separation of indigenous noroviruses from sewage sludge and treated wastewater

    D. Sano, Y. Ueki, T. Watanabe, T. Omura

    WATER SCIENCE AND TECHNOLOGY　54　(3)　77-82　2006年

    出版者・発行元：I W A PUBLISHING

    DOI： 10.2166/wst.2006.451 　

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    In this study, feasibility of membrane separation for the removal of indigenous noroviruses (NVs) is evaluated. The indigenous NV gene was never detected from ultrafiltration (UF) permeates of sewage sludge and treated wastewater. Indigenous NV gene was also not detected from permeates of sewage sludge and treated wastewater by microfiltration (IVIF) with a pore size of 0.1 mu m (MF0.1). Even though the pore size of IVIF (0.1 mu m) was much larger than the diameter of virus particle (approximately 30-40 nm), more than 4-log(10) reduction value (LRV) at maximum was achieved by membrane separation with MF0.1. NV genes were often detected from permeates of sewage sludge and treated wastewater by IVIF with a pore size of 0.45 mu m (MF0.45), although the maximum log(10) reduction values were more than 3.59 for sewage sludge and more than 2.90 for treated wastewater. It is important to verify factors determining the removal efficiency of viruses With MF membranes.

  50. Norovirus pathway in water environment estimated by genetic analysis of strains from patients of gastroenteritis, sewage, treated wastewater, river water and oysters

    Y Ueki, D Sano, T Watanabe, K Akiyama, T Omura

    WATER RESEARCH　39　(18)　4271-4280　2005年11月

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1016/j.watres.2005.06.035 　

    ISSN：0043-1354

    詳細を見る 詳細を閉じる

    In this study, Norovirus (NV) capsid gene was detected from patients of gastroenteritis, domestic sewage, treated wastewater, river water and cultivated oysters in geographically close areas where all of samples were collected. In order to improve recovery efficiency of NVs from oysters, a new method using a spallation apparatus was developed. As a result, 18 of 30 oysters (60%) were positive for NV gene, while 7 of 30 (23%) oysters from the same sampling point were positive with the conventional ultracentrifugal method between November 2003 and February 2004. These results indicate that our new method exhibits the higher efficiency of recovering NVs than the conventional ultracentrifugal method. Six of 8 samples (75%) of river water were positive for NV gene between November 2003 and February 2004. Further-more, 8 of 9 samples (89%) of treated wastewater and all 9 samples of sewage were positive for NV gene in the same period. These results indicated that treated wastewater would be one of the main sources for NV pollution in this area. The phylogenctic analysis in isolated NV capsid genes was conducted, in which high identities of gene sequences between NVs from patients, domestic sewage, river water and cultivated oysters were observed. These results implied that there would be a geographically associated circulation of NVs between human and cultivated oysters via water environment. It would be important to quantitatively analyze the moving pathway of NVs, which directly link to the development of a new scheme for preventing water environment and cultivated oysters from NV contamination. (c) 2005 Elsevier Ltd. All rights reserved.

  51. Construction of a cloning system for the mass production of a virus-binding protein specific for poliovirus type 1

    Daisuke Sano, Tatsuo Omura

    Applied and Environmental Microbiology　71　(5)　2608-2615　2005年5月

    DOI： 10.1128/AEM.71.5.2608-2615.2005 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    In our previous study, virus-binding proteins (VBPs) demonstrating the ability to strongly bind poliovirus type 1 (PV1) were recovered from a bacterial culture derived from activated sludge. The isolated VBPs would be useful as viral adsorbents for water and wastewater treatments. The VBP gene of activated sludge bacteria was isolated, and the cloning system of the VBP was established. The isolation of the VBP gene from DNA libraries for activated sludge bacteria was achieved with the colony hybridization technique. The sequence of the VBP gene consisted of 807 nucleotides encoding 268 amino acids. Fifteen amino acid sequences were retrieved from 2,137,877 sequences by a homology search using the BLAST server at the National Center for Biotechnology Information. The protein encoded in the isolated genome was considered to be a newly discovered protein from activated sludge culture, because any sequences in protein databases were not perfectly matched with the sequence of the VBP. It was confirmed that Escherichia coli BL21 transformed by pRSET carrying the isolated VBP gene could extensively produce the VBP clones. Enzyme-linked immunosorbent assay (ELISA) revealed that the VBP clone exhibited the binding ability with intact particles of PV1. The equilibrium binding constant between PV1 and VBP in the ELISA well was estimated to be 2.1 × 107 (M-1), which also indicated that the VBP clones have a high affinity with the PV1 particle. The VBP cloning system developed in this study would make it possible to produce a mass volume of VBPs and to utilize them as a new material of the specific adsorbent in several technologies, including virus removal, concentration, and detection. Copyright © 2005, American Society for Microbiology. All Rights Reserved.

  52. ウイルスの分析及び膜分離活性汚泥法によるウイルス除去性能の評価

    大村 達夫, 渡部 徹, 佐野 大輔, 植木 洋

    財団法人建設工学研究振興会年報　40　49-55　2005年4月1日

    出版者・発行元：東北大学

    ISSN：0288-9994

  53. 膜処理による下水処理水中のSRSV除去研究

    大村 達夫, 渡部 徹, 佐野 大輔, 熊谷 幸博

    財団法人建設工学研究振興会年報　40　42-48　2005年4月1日

    出版者・発行元：東北大学

    ISSN：0288-9994

  54. Virus-binding proteins recovered from bacterial culture derived from activated sludge by affinity chromatography assay using a viral capsid peptide

    D Sano, T Matsuo, T Omura

    APPLIED AND ENVIRONMENTAL MICROBIOLOGY　70　(6)　3434-3442　2004年6月

    出版者・発行元：AMER SOC MICROBIOLOGY

    DOI： 10.1128/AEM.70.6.3434-3442.2004 　

    ISSN：0099-2240

    詳細を見る 詳細を閉じる

    The contamination of water environments by pathogenic viruses has raised concerns about outbreaks of viral infectious diseases in our society. Because conventional water and wastewater treatment systems are not effective enough to inactivate or remove pathogenic viruses, a new technology for virus removal needs to be developed. In this study, the virus-binding proteins (VBPs) in a bacterial culture derived from activated sludge were successfully recovered. The recovery of VBPs was achieved by applying extracted crude proteins from a bacterial culture to an affinity column in which a custom-made peptide of capsid protein from the poliovirus type 1 (PV1) Mahoney strain (H2N-DNPASTTNKDKL-COOH) was immobilized as a ligand. VBPs exhibited the ability to adsorb infectious particles of PV1 Sabin I as determined by enzyme-linked immunosorbent assay. The evaluation of surface charges of VBPs with ion-exchange chromatography found that a majority of VBP molecules had a net negative charge under the conditions of affinity chromatography. On the other hand, a calculated isoelectric point implied that the viral peptide in the affinity column was also charged negatively. As a result, the adsorption of the VBPs to the viral peptide in the affinity column occurred with a strong attractive force that was able to overcome the electrostatic repulsive force. Two-dimensional electrophoresis revealed that the isolated VBPs include a number of proteins, and their molecular masses were widely distributed but smaller than 100 kDa. Amino acid sequences of N termini of five VBPs were determined. Homology searches for the N termini against all protein sequences in the National Center for Biotechnology Information (NCBI) database showed that the isolated VBPs in this study were newly discovered proteins. These VBPs that originated with bacteria in activated sludge might be stable, because they are existing in the environment of wastewater treatments. Therefore, a virus removal technology utilizing VBPs as viral adsorbents can be developed, since it is possible to replicate VBPs by protein cloning techniques.

  55. ウイルスの分析調査

    大村 達夫, 渡部 徹, 佐野 大輔, 熊谷 幸博

    財団法人建設工学研究振興会年報　39　61-67　2004年4月1日

    出版者・発行元：東北大学

    ISSN：0288-9994

  56. Detection of infectious pathogenic viruses in water and wastewater samples from urbanised areas

    D Sano, T Watanabe, T Matsuo, T Omura

    WATER SCIENCE AND TECHNOLOGY　50　(1)　247-251　2004年

    出版者・発行元：I W A PUBLISHING

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Regardless of the remarkable improvement in sanitary conditions in urbanised areas, infectious diseases caused by pathogenic viruses in water have been reported year after year. The actual situation for the behaviour of pathogenic viruses in urbanised society should be understood. In this study, infectious enteroviruses and adenoviruses in water and wastewater samples from urbanised areas were investigated. Infectious enteroviruses were continuously detected from sewage and sewage sludge during the summer season. The detection of infectious viruses in sewage and sewage sludge was facilitated by the enzymatic virus elution (EVE) method. The concentration of infectious enteroviruses in positive samples of sewage and sewage sludge was &gt;400 virions/L. Infectious viruses were also detected from water samples at levels &gt;4 virions/L.

  57. Affinity isolation of algal organic matters able to form complex with aluminium coagulant

    T Takaara, D Sano, H Konno, T Omura

    4TH WORLD WATER CONGRESS: INNOVATION IN DRINKING WATER TREATMENT　4　(5-6)　95-102　2004年

    出版者・発行元：I W A PUBLISHING

    ISSN：1606-9749

    詳細を見る 詳細を閉じる

    Mass propagation of algae in drinking water source affects the coagulation process in water treatment systems. Many researchers indicated that some kinds of algal organic matters (AOMs) interacted with coagulants by coordinate bonds and brought about several disorders due to the increase in both the coagulant demand and the residual coagulant in treated water. However, the inhibitory mechanism on the coagulation by AOMs has not been fully elucidated. In this study, AOMs able to form complex with aluminium coagulant were isolated and analyzed. The results of the gel chromatography showed that extracellular organic matters (EOMs) from Microcystis aeruginosa (NIES-91) have molecular weight of between 10 and 20 kDa, around 40 kDa and more than 600 kDa. The amount and diversity of EOMs increased as the algal growth. AOMs able to form complex with aluminium coagulant were successfully isolated with the affinity chromatography. SDS-PAGE analysis revealed that these AOMs included proteins that have molecular weight between 43 and 67 kDa. Since several kinds of proteins such as metallothionein are known for strongly adsorbing multivalent cations, the isolated algal proteins able to form complex with aluminium might have a high capacity of capturing coagulants and inhibit the coagulation in the drinking water treatment.

  58. Detection of enteric viruses in municipal sewage sludge by a combination of the enzymatic virus elution method and RT-PCR

    D Sano, K Fukushi, Y Yoshida, T Omura

    WATER RESEARCH　37　(14)　3490-3498　2003年8月

    出版者・発行元：PERGAMON-ELSEVIER SCIENCE LTD

    DOI： 10.1013/S0043-1354(03)00208-2 　

    ISSN：0043-1354

    詳細を見る 詳細を閉じる

    Pathogenic enteric viruses can be retained in municipal sewage sludge as has been reported by many researchers. Although the RT-PCR technique has been extensively employed for the virus detection from various environmental samples, the application of RT-PCR to the detection of viruses in sewage sludge has the difficulty because of inhibitory substances to the gene amplification. However, a combination of the enzymatic virus elution (EVE) method with RTPCR made it possible to effectively detect viruses in sewage sludge. The enzymatic breakdown of sludge flocs in the EVE method enhanced the virus elution from poliovirus 1 (PV1)-inoculated sewage sludge, and the detection of PV1 was performed by RT-PCR without any inhibitions. On the contrary, the application of RT-PCR to the viral assay in the USEPA method using the 10% beef extract solution was not practical because of inhibitions to the viral gene amplification. The combination of the EVE method using lysozyme (polysaccharide-degrading enzyme), papain (protease), and chymotrypsin (protease) with RT-PCR resulted in a virus recovery efficiency of 31%, but a synergistic effect of these enzymes on the virus recovery efficiency was not observed. The EVE method using lysozyme or papain could be a promising procedure for the virus elution from sewage sludge in detecting these viruses with RT-PCR. (C) 2003 Elsevier Science Ltd. All rights reserved.

  59. ウサギ顎関節円板切除および下顎頭切除後の同種骨移植による下顎頭再建に関する研究

    佐野 大輔

    愛知学院大学歯学会誌 = The Aichi-Gakuin journal of dental science　41　(1)　21-34　2003年3月31日

    出版者・発行元：愛知学院大学歯学会

    ISSN：0044-6912

  60. Functional metal-binding proteins by metal-stimulated bacteria for the development of an innovative metal removal technology

    T Antsuki, D Sano, T Omura

    WATER SCIENCE AND TECHNOLOGY　47　(10)　109-115　2003年

    出版者・発行元：I W A PUBLISHING

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Heavy metal pollution has become an environmental problem throughout the world because heavy metals can be accumulated into the food chain and bring about serious problems, not only for ecosystems but also for human health. In this study, functional metal-binding proteins (FMBPs) were isolated from a metal-stimulated activated sludge culture with the aim of applying them to an innovative metal removal technology. Activated sludge bacteria was cultured in growth media including copper ion, and the stimulation of protein production by copper ion led to the 14% increase in a quantity of extracted crude proteins per 1 g of bacterial cell pellet (wet). In order to isolate FMBPs,. extracted crude proteins were. applied to the immobilized metal affinity column in which each of copper, nickel and zinc was used as a ligand. Several FMBPs were succesfully isolated from copper-stimulated bacteria. One of FMBPs. (molecular weight of about 40 kDa) exhibited an ability to adsorb all three metals. The multi metal-binding property of this FMBP could be applied to an innovative metal removal technology: Furthermore, isolated. FMBPs that could capture only one kind of heavy metal would also be attractive as a metal adsorbent in recovering a specific metal as a resource from wastewater, including several heavy metals.

  61. Risk evaluation for pathogenic bacteria and viruses in sewage sludge compost

    T Watanabe, D Sano, T Omura

    WATER SCIENCE AND TECHNOLOGY　46　(11-12)　325-330　2002年

    出版者・発行元：I W A PUBLISHING

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    Composting can be regarded as the most available option for recycling of sewage sludge. However, the existence of pathogenic bacteria and viruses in the compost has been scarcely investigated until now: So there is little information on the infectious risk through agricultural activities or gardening in using the compost. In this study, several kinds of composts were investigated for detection of pathogenic bacteria (Salmonella spp. and Escherichia coli O157) and enteric viruses: It was concluded from the result that these bacteria and viruses could not be detected in 1.0 g-wet of any kinds of composts. Infectious risks through agricultural activities or gardening were evaluated by Monte Carlo simulation in the case that the compost was polluted by Salmonella spp., E. coli O157:117 and Poliovirus 1. Criteria satisfying the acceptable risk (less than 10(-4) per year) for these pathogenic bacteria and virus in the compost were determined from the result of simulations. 1.0 [CFU or PFU/g-wet] was available as the criteria for E. coli O157 and Poliovirus 1 in the compost. On the other hand, the criterion for Salmonella spp. in the compost should be established on a lower concentration than 0.001 CFU/g-wet.

  62. Enhanced virus recovery front municipal sewage sludge with a combination of enzyme and cation exchange resin

    D Sano, K Fukushi, K Yano, Y Yoshida, T Omura

    WATER SCIENCE AND TECHNOLOGY　43　(2)　75-82　2001年

    出版者・発行元：I W A PUBLISHING

    ISSN：0273-1223

    詳細を見る 詳細を閉じる

    There is a great difficulty in virus enumeration in sewage sludge because viruses in sludge are firmly captured by sludge solids. In order to determine the precise number of viruses in sludge, an enhanced virus recovery method with a combination of an enzyme and a cation exchange resin (CER) was developed. Test viruses were seeded to a sample sludge obtained from a municipal wastewater treatment plant, and the sludge were incubated with various eluents. The quantity of eluted viruses in the liquid phase was then measured by the plaque assay technique. Using the eluent containing only water, CER, and CER with enzyme exhibited 0 %, 19 % and 39 % of virus recovery, respectively. While the conventional USEPA method exhibited a virus recovery of 21 %. Furthermore, viruses eluted by the eluent containing the CER and the lysozyme included not only surface-attached viruses but also solids-embedded viruses.

  63. 酵素・キレーター混合液を用いた下水汚泥からの腸管系ウイルス誘出

    佐野大輔, 福士謙介, 大村達夫

    日本水環境学会年会講演集　33rd　1999年

  64. 下水汚泥からのウイルス誘出における多糖分解酵素の有効性

    佐野大輔, 福士謙介, 熊谷幸博, 大村達夫

    土木学会年次学術講演会講演概要集 第7部　53rd　1998年

︎全件表示 ︎最初の5件までを表示

書籍等出版物 3

  1. 環境と微生物の事典

    2014年

  2. 水再生利用学

    浅野孝, 大垣眞一郎, 江藤隆, 滝沢智, 船水尚行, 松井正樹, 田中宏明

    技法堂出版　2010年

  3. Virus-binding proteins in water environments: natural ligands for human viruses, p. 257-272. In George V. Kurladze (ed.), Environmental Microbiology Research Trends.

    Daisuke Sano, Tatsuo Omura

    Nova Science Publishers Inc., Hauppauge, N.Y.　2008年

講演・口頭発表等 48

  1. Diverse enteric bacteria can excrete histo-blood group antigens as adsorbents for human gastroenteritis viruses 国際会議

    The 63rd Annual Meeting of the Japanese Society for Virology　2015年11月

  2. Estimation Accuracy of Pathogen Density Distribution in Water Based on a Left-Censored Dataset: Bayesian vs Maximum a Posteriori 国際会議

    IEEC2015　2015年10月

  3. 衛生環境工学におけるゲノム解析技術の応用 招待有り

    佐野大輔

    第158回日本獣医学会学術集会・公衆衛生学分科会シンポジウム「ゲノム解析技術が生み出す獣医公衆衛生学領域における新知見紹介」　2015年9月7日

  4. Diversity of histo-blood group antigen-positive enteric bacteria in water: possible environmental vehicle for human norovirus 国際会議

    WaterMicro2015　2015年9月

  5. 下水再生利用に関わる下水処理装置の性能評価及び運転管理手法の提案 招待有り

    佐野大輔

    7. 土木学会環境工学委員会・次世代下水道小委員会講演会　2015年8月11日

  6. 胃腸炎ウイルスの特性と水系感染リスク制御 招待有り

    佐野 大輔

    京都大学大学院工学研究科・環境微生物学特論　2015年7月7日

  7. Genetic variations of norovirus GII.4 within a single host and among hosts 国際会議

    Special Workshop on Health-Related Environmental Virology　2015年5月27日

  8. 胃腸炎ウイルスの感染制御因子 招待有り

    佐野大輔

    第11回小児消化管感染症研究会　2015年2月7日

  9. 胃腸炎ウイルスの環境中動態を含めた生態の解明に向けて

    佐野 大輔

    5. 第11回小児消化管感染症研究会　2014年11月9日

  10. Bayesian modeling of virus removal efficiency in wastewater treatment processes 国際会議

    IWA World Water Congress & Exhibition　2014年9月

  11. Estimation of quantitative relationships between pathogens and indicator microorganisms in water using left-censored data 国際会議

    ISFEV2014　2014年9月

  12. Development of a screening assay of norovirus-binding enteric bacteria bearing histo-blood group antigen-like substances 国際会議

    IUMS2014　2014年7月

  13. Research trends and future perspectives of environmental and public health virology 国際会議

    The 4th Young Professors and Students Symposium　2014年3月27日

  14. Norovirus-binding enteric bacteria: Significance in the environmental dissemination of gastroenteritis viruses 国際会議

    The 2nd Global Leadership Initiative Special Workshop on Water Virology　2014年1月

  15. Stability of rotavirus infectivity analyzed by oxidative stress marker detection 国際会議

    The 5th European Rotavirus Biology Meeting　2013年10月

  16. ノロウイルス〜MBRにおける除去性、感染リスク管理、及び性能基準 招待有り

    佐野 大輔

    3. 平成25年度先端膜工学研究推進機構秋季講演会・膜工学サロン　2013年9月10日

  17. A hidden relationship between norovirus and enteric bacteria: norovirus exploits histo-blood group antigen-like substances on bacterial cells for its better survival in nature? 国際会議

    The 47th Joint Working Conference on Viral Diseases, US-Japan Cooperative Medical Science Program　2013年3月13日

  18. Development of a novel photocatalytic nano-composite membrane to create new water resources from wastewater 国際会議

    SJ-NANO2013 (Spanich-Japanese Bilateral Workshop)　2013年3月5日

  19. Performance target of enteric virus removal with MBR 国際会議

    Global Leadership Initiative Special Workshop on Water Virology　2013年1月19日

  20. Multiple peak expression of enteric virus density in environmental water 国際会議

    佐野 大輔

    The 4th Asia-Pacific Young Water Professionals Conference 2012　2012年12月

  21. Infectivity of gastroenteritis viruses analyzed by oxidative stress marker detection and RNase sensitivity test 国際会議

    ISFEV2012　2012年10月

  22. Human norovirus-binding enteric bacteria bearing histo-blood group antigen-like extracellular polymeric substances 国際会議

    The 46th Joint Working Conference on Viral Diseases, US-Japan Cooperative Medical Science Program　2012年6月19日

  23. Accumulation of oxidative damages on rotavirus capsid protein and its relationship with infectivity 国際会議

    Daisuke Sano

    The 2nd COST 929 Symposium, Istanbul, Turkey　2010年10月7日

  24. Human enteric bacteria that capture norovirus particles with a specific interaction through histo-blood group antigen-like moiety 国際会議

    Daisuke Sano

    The 2nd COST 929 Symposium, Istanbul, Turkey　2010年10月7日

  25. Norovirus-binding bacteria: significance in the environmental dissemination of gastroenteritis viruses 国際会議

    Daisuke Sano

    The 44th Joint Working Conference on Viral Diseases, US-Japan Cooperative Medical Science Program, Sapporo, Japan, June 28-29.　2010年

  26. Detection of oxidative damages on capsid proteins of norovirus: a new approach to evaluate the infectivity of non-culturable virus

    Daisuke Sano

    The 15th Symposium on Health-Related Water Microbiology　2009年

  27. Waterborne viral gastroenteritis - could prevention be within our reach? 国際会議 招待有り

    Daisuke Sano

    Academie Des Sciences-JSPS Workshop on Environment and Health Challenges, Paris, France. May 28-29.　2009年

  28. A new approach to evaluate the infectivity of non-cultivatable viruses 国際会議

    Daisuke Sano

    The 43rd Joint Working Conference on Viral Diseases, US-Japan Cooperative Medical Science Program, Philadelphia, Pennsylvania, USA, July 20-22.　2009年

  29. 培養できない腸管系ウイルス不活化評価を目的とした外殻タンパク質酸化傷害検出手法の開発

    佐野大輔

    第46回環境工学研究フォーラム　2009年

  30. New detection method for enteric viruses in digestive diverticulum of Pacific oyster, Crassostrea gigas utilizing enzymes for virus elution

    The 15th Symposium on Health-Related Water Microbiology　2009年

  31. Quantification and molecular characterization of norovirus, sapovirus and hepatitis A virus in the Llobregat river basin (Catalonia, NE Spain)

    The 15th Symposium on Health-Related Water Microbiology　2009年

  32. Genetic diversity and evolution in amino acid sequences of norovirus genogroup II strains in environmental and clinical samples

    The 15th Symposium on Health-Related Water Microbiology　2009年

  33. Research activities on waterborne pathogenic viruses in Japan 国際会議 招待有り

    Daisuke Sano

    Seminar of molecular microbiology and virology　2007年6月26日

  34. Quantification of human astroviruses using a one-step real-time quantitative RT-PCR 国際会議

    Daisuke Sano

    The 14th Symposium on Health-Related Water Microbiology (WaterMicro07)　2007年

  35. Affinity isolation of norovirus-binding proteins from activated sludge microorganisms using a surface-exposed part of viral capsid protein

    The 14th Symposium on Health-Related Water Microbiology (WaterMicro07)　2007年

  36. 水中病原ウイルスの新規検出技術に用いる固定化ウイルス吸着タンパク質の活性評価

    平成17年度土木学会東北支部技術研究発表会　2006年

  37. ファージディスプレイ法を用いた凝集阻害誘因タンパク質をコードする藻類遺伝子の分離

    第43回環境工学研究フォーラム　2006年

  38. Seasonal variation of Norovirus occurrence in water environment

    The 5th World Water Congress of International Water Association　2006年

  39. 活性汚泥細菌ゲノムDNAライブラリから取得した新規重金属吸着タンパク質遺伝子

    第42回環境工学研究フォーラム　2005年

  40. Heavy metal-binding proteins from metal-stimulated bacteria as a novel adsorbent for metal removal technology 国際会議

    Daisuke Sano

    The 1st IWA-ASPIRE Conference & Exhibition　2005年

  41. Membrane separation of indigenous Norovirus from sewage sludge and treated wastewater 国際会議

    Daisuke Sano

    The 13th Symposium on Health-Related Water Microbiology (WaterMicro05)　2005年

  42. アフィニティクロマトグラフィによる活性汚泥細菌からのアデノウイルス吸着タンパク質（Adenovirus-Binding Protein: ADVBP）の分離

    第59回土木学会年次学術講演会　2004年

  43. アデノウイルス吸着タンパク質（Adenovirus-Binding Protein: ADVBP）の活性汚泥細菌からの分離

    第41回環境工学研究フォーラム　2004年

  44. 活性汚泥細菌から分離されたウイルス吸着タンパク質とポリオウイルス粒子間の相互作用評価

    平成14年度土木学会東北支部技術研究発表会　2003年

  45. Detection of infectious pathogenic viruses in water and wastewater samples from urbanized area

    The 12th Symposium on Health-Related Water Microbiology　2003年

  46. Functional Metal-Binding Proteins by Metal-stimulated Bacteria for the Development of an Innovative Metal Removal Technology 国際会議

    Daisuke Sano

    The 3rd World Water Congress of International Water Association　2002年

  47. Enhanced virus recovery from municipal sewage sludge with the combination of enzyme and cation exchange resin 国際会議

    Daisuke Sano

    The 1st World Water Congress of International Water Association　2000年

  48. Enumeration of enteric viruses in municipal sewage sludge with enzymatic virus elution method. 国際会議

    Daisuke Sano

    Asian Waterqual '99, 7th IAWQ Asia-Pacific Regional Conference　1999年

︎全件表示 ︎最初の5件までを表示

共同研究・競争的資金等の研究課題 42

  1. 水中病原体群と指標群の相関解析アルゴリズムの開発

    加藤 毅, 佐野 大輔

    2022年4月1日 ～ 2026年3月31日

  2. 新型コロナ様パンデミックを誘因する未知＆変異病原ウイルスの下水からの早期検出手法

    大村 達夫, 佐野 大輔, 渡辺 幸三

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (B)

    研究機関：Tohoku University

    2022年4月1日 ～ 2025年3月31日

  3. 脱炭素化に向けた創エネ・リン資源回収型下水高度処理システムの開発

    李 玉友, 北條 俊昌, 安井 英斉, 増田 周平, 久保田 健吾, 佐野 大輔, 覃 宇

    2022年4月1日 ～ 2025年3月31日

  4. 下水疫学を用いたアジアの途上国における病原ウイルス監視基盤の構築と感染リスク制御

    福士 謙介, 渡部 徹, 佐野 大輔, 渡辺 幸三, 大村 達夫

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (A)

    研究機関：The University of Tokyo

    2022年4月1日 ～ 2025年3月31日

  5. 都市下水から薬を創り出す：棄てられたヒト免疫細胞由来遺伝子を活用した抗体医薬開発

    佐野 大輔, 野地 智法, 斉藤 繭子, 久保田 健吾

    2021年7月9日 ～ 2025年3月31日

    詳細を見る 詳細を閉じる

    本研究は、都市下水から回収した抗体遺伝子を用いて抗体医薬の合成を行う手法の開発に取り組むものである。研究初年度である令和3年度においては、文献情報をもとに免疫細胞由来遺伝子を増幅するためのプライマーを設計し、都市下水から抽出した総DNAを鋳型として用いたPCRを行い、都市下水から免疫細胞由来遺伝子の増幅産物を取得することを試みた。都市下水は3000xgで30分遠心分離し、得られた沈殿物を下水濃縮サンプルとした。プライマーは複数の組み合わせを合成し、試行錯誤的に下水濃縮サンプルから抽出した総DNAに適用した。その結果、目的遺伝子と思われる増幅産物を得ることに成功した。得られた増幅産物に対して配列解析を行ったところ、ヒト免疫グロブリン由来の遺伝子配列が得られていることが確認された。この結果により、都市下水中に免疫細胞由来遺伝子が存在していることが証明され、本研究のコンセプトが成立可能であることが示された。続いて、得られた増幅産物に対して次世代シーケンシングを行うための前処理を行なった。最終的に、十分な量と純度を持つサンプルが得られたため、次世代シーケンス解析を開始した。

  6. 水の消毒処理に対するウイルスの耐性―遺伝的要因の解明による安全な水利用の実現―

    佐野 大輔, 片山 浩之, 片山 和彦, 北島 正章

    2021年4月5日 ～ 2025年3月31日

    詳細を見る 詳細を閉じる

    研究初年度である令和3年度においては、世界中で水の消毒に使用されている塩素消毒について、感受性の異なるロタウイルス集団を計40集団取得し、塩素消毒実験結果をもとに、17の低感受性（高抵抗性）集団、及び23の高感受性（低抵抗性）集団に分割した。これらのロタウイルス集団に対して次世代シーケンシングによる全ゲノム解析を行ったところ、低感受性集団に特徴的な遺伝子変異は存在しないことが明らかとなった。そこで低感受性集団と高感受性集団の間で遺伝的多様性を比較したところ、外殻タンパク質VP7のアミノ酸配列において、低感受性集団の方が高感受性集団よりも有意に多様性が高いことが判明した。遺伝的多様性が高い場合に集団内で粒子間の凝集が生じやすくなり、それが消毒耐性をもたらすとの仮説を立て、進化シミュレーションにより再現を試みたところ、凝集しやすい集団ほど、消毒耐性がランダムに変動しつつ遺伝的多様性が高く保たれることが明らかとなった。この結果から、代表的な胃腸炎ウイルスであるロタウイルスに関しては、遺伝的多様性自体が集団として消毒剤への低感受性を示す理由であることが示唆された。

  7. スリランカにおける下水水質情報に基づいた下水管路劣化予測システムの開発

    佐野 大輔, 水谷 大二郎, Amarasiri Mohan

    2021年10月7日 ～ 2024年3月31日

    詳細を見る 詳細を閉じる

    本研究は、スリランカにおいて、微生物の起源を同定する手法であるMicrobial Source Tracking（MST）の手法を応用し、下水管の破損部分から混入する土壌由来の微生物の下水中に占める割合を空間的・時間的に把握することで、下水管破損箇所を推定し、かつ下水管劣化速度の予測に適用するための新規手法を開発することを目指すものである。研究初年度である令和3年度においては、まずスリランカ側カウンターパート（チャンディカ・ガマゲ（ペラデニヤ大学医学部・講師）、及びチャミンダ・サマラスーリヤ（モラトゥワ大学地球資源工学部・講師））とのオンライン打ち合わせを通じ、スリランカ国内における調査地点の確定を行った。希望する調査地点で下水採取を行うために、所轄官庁向けの要望書を作成・提出したところである。さらに、本研究で想定している手法が適用可能であることを確認するために、日本国内で未処理下水を採取し、総DNAを抽出して、細菌の分類に用いられる16s rRNA遺伝子配列の取得を行った。その結果、下水に特徴的な細菌種の16s rRNA遺伝子配列が確実に得られることを確認することに成功し、スリランカで得られたサンプルに適用する準備が整えられた。

  8. 水質変換効率を予測するための新規光化学反応モデルの構築

    吉村 千洋, 佐野 大輔

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (B)

    研究機関：Tokyo Institute of Technology

    2021年4月1日 ～ 2024年3月31日

  9. 水環境に潜伏する薬剤耐性菌から人への耐性遺伝子の伝播機構とリスク評価

    鈴木 聡, 佐野 大輔, 渡辺 幸三

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (A)

    研究機関：Ehime University

    2020年4月1日 ～ 2024年3月31日

    詳細を見る 詳細を閉じる

    本研究では，環境に潜伏する耐性遺伝子の実態解明からさらに進めて，水環境に存在する陸起源および水圏起源の耐性遺伝子が人獣病原菌・腸内細菌に伝播するプロセスを解析し，水圏から人間環境へ耐性遺伝子が侵入する機構・条件の解明とリスク評価を目的としている。動物・ヒト間での遺伝子伝播の研究はあるが，自然水圏からヒトへのリンク機構を解明するのは新規研究であり，ワンヘルスに環境視点で貢献する研究である。 これまでに，水環境での耐性遺伝子と伝達因子の相関解析を行い，比較的新しく見つかった耐性遺伝子は多様な伝達因子に担われるが，徐々に特定のものに収束することが示唆された。また，アルファプロテオバクテリアに属する海洋細菌に多く見出される新規のsul4は，非伝達性であり，通常の葉酸合成系で働く酵素であることがわかった。この遺伝子は河川の細菌では伝達性が疑われていることから（既報），海洋細菌と河川細菌では機能が変化していることが示唆される。今後の詳細な解析が望まれる。 下水環境と海洋環境から得た耐性菌の持つ耐性遺伝子の伝達を実験的に検討し，既報より高率に伝達が起こりうることを明らかにした。遺伝子供与菌の栄養状態が低下すると伝達性も低下し，有機物添加で伝達率が回復することから，有機物濃度の高い下水中では，遺伝子伝播が起こりやすいことが考えられた。 また，化学物質の共存などで伝達率が上昇する条件を多くの既報から抽出し，決定木解析で明確にしつつある。耐性遺伝子が多様な細菌群集へ拡散する機構と条件の解明が期待できる。

  10. 下水中のA型・E型肝炎ウイルスおよび無承認無許可医薬品の実態把握とリスク評価

    秋葉 道宏, 小坂 浩司, 三浦 尚之, 佐野 大輔

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (B)

    研究機関：National Institute of Public Health

    2020年4月1日 ～ 2023年3月31日

    詳細を見る 詳細を閉じる

    2021年度は，人口規模の異なる二つの都市の下水処理場において流入下水試料を原則2週に1度の頻度で採水し，A型およびE型肝炎ウイルス，新型コロナウイルスの調査を継続した。そのうち，一つの都市で収集した試料では，下水試料を陰電荷膜でろ過することにより水中の固形分と共にウイルス粒子を膜に吸着させ，膜から直接ウイルスRNAを抽出・精製する手法を検討した。この手法では，A型およびE型肝炎ウイルスは不検出だったが，新型コロナウイルスRNAは検出された。新型コロナウイルスRNAの検出に，５つのリアルタイムRT-PCRアッセイ（CDC_N1，N2，N3，Sarbeco，NIID系）を比較した結果，CDC_N2系による検出感度が最も高かった。本研究で検討した手法により，新型コロナウイルス感染症の流行の第４波から第６波に対応して，下水試料中の新型コロナウイルスRNA濃度および検出率が変動することが示された。 無承認無許可医薬品に該当する製品から検出事例がある医薬品成分を含めた200以上の化学物質について，6種の下水試料を対象にLC-Orbitrap/MSを用いてターゲットスクリーニング分析を行った。無承認無許可医薬品に含まれる7種の医薬品成分（シブトラミン，シルデナフィル，タダラフィル，フェンテルミン，ブホテニン，フルオキセチン，ヨヒンビン）は検出されなかったが，下水試料から15～20種程度の物質が10 ng/L以上の濃度で検出された。検出された主な物質は医薬品類であったが，それ以外には農薬，人工甘味料，工業用化学物質も検出され，μg/Lの濃度で検出された物質もあった。

  11. 機械学習による水中病原体と指標微生物の濃度相関解析法の精緻化

    加藤 毅, 佐野 大輔

    2019年4月1日 ～ 2023年3月31日

    詳細を見る 詳細を閉じる

    環境水や飲用水には感染症を引き起こす病原体が含まれることがある．病原体とは病原細菌，病原ウイルス，および原虫を含む．モニタリングされる指標微生物の水質衛生基準値を設定する際には，指標微生物濃度と病原体濃度との相関関係を適切に定量化することが求められる．しかし，病原体の陽性率の低さが相関計算の障害となっている．本研究では，近年著しい発展を遂げる機械学習を使うことで，指標微生物と病原体の濃度相関解析を改善することを目指した．従来は指標微生物や病原体の濃度を使って相関を計算してきた．これに対し，本研究は，指標微生物や病原体の濃度と同時に取得できる水質データやドメイン知識を利活用することで，相関解析法の高精度化を狙った． 統計学において，トビット法という，非定量値と回帰直線を同時に推定する方法がある．トビット法では，観測された病原体濃度の確率密度関数，および検出限界を下回った濃度がその確率モデルにおいて非観測となる確率質量関数を得ることが出来，それらを組み合わせることで回帰係数の尤度関数を構成する．このアプローチをそのまま使ってしまう場合，予測に用いる情報が不十分になるため，非検出値の予測分布は大雑把なものにしかならず，そこから得られる相関解析は十分な精度で得られない．本研究では，水質データの援用によって，非検出値の予測分布を精密にすることで，相関係数の精度を向上させた．個別の説明変数の予測能力はさほど強くないため，標本が小さいとき符号が逆転した標本相関が発生する．本研究では，ドメイン知識を符号制約で表すことで，標本が小さくても，回帰分析の精度を大きく改善させ，これを相関解析のワークフローに取り込むことで，相関解析の精度を向上させることに成功した．

  12. 下水再生利用におけるエネルギー回収と健康リスク管理の実現

    提供機関：Japan Science and Technology Agency (JST)

    制度名：Strategic International Collaborative Research Program

    研究機関：Tohoku University

    2019年4月 ～ 2022年3月

  13. サニテーション価値連鎖の提案 －地域のヒトによりそうサニテーションのデザイン－

    提供機関：Research Institute for Humanity and Nature

    制度名：Research Project

    研究機関：Research Institute for Humanity and Nature

    2015年4月 ～ 2022年3月

  14. 水処理を目的とした自己再生能力の高いナノカーボン・二酸化チタン複合材料の新規開発

    吉村 千洋, 森 伸介, 佐野 大輔, クルニアワン ウィナルト, 日野出 洋文

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (B)

    研究機関：Tokyo Institute of Technology

    2018年4月1日 ～ 2021年3月31日

    詳細を見る 詳細を閉じる

    難分解性有機化合物や病原微生物の除去を目的として、ナノカーボン材料の簡易生成法の開発、またその金属触媒との複合材料の最適化および反応特性の解明を実施した。さらに、最適化した磁性カーボンナノチューブと酸化チタンの複合材料について、下水の二次処理水での除去特性を調べ、難分解性有機化合物および微生物の吸着・光触媒分解の解明とモデル化を行った。

  15. ノロウイルスの水環境中キャリアーとなる糖鎖分泌細菌の多様性と存在実態の解明 競争的資金

    佐野 大輔

    2017年4月 ～ 2021年3月

  16. 衛生安全計画に基づいた下水処理放流水質の衛生工学的管理スキームの構築

    2018年10月 ～ 2021年2月

  17. なぜノロウイルスがカキに蓄積するのか？そのメカニズムの解明

    大村 達夫, 藤井 学, 三浦 尚之, 渡辺 幸三, 佐野 大輔

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Challenging Research (Pioneering)

    研究機関：Tohoku University

    2017年6月30日 ～ 2020年3月31日

    詳細を見る 詳細を閉じる

    本研究は、沿岸域のプランクトンが環境からカキにノロウイルスを運んでいるという仮説を立てて、その検証を行った。カキを養殖している松島湾内の複数地点において動植物プランクトンとカキを採取した。動物プランクトンと植物プランクトンを顕微鏡下の形態観察に基づいてソーティングし、それぞれからノロウイルス遺伝子を検出および定量した結果、一部サンプルからノロウイルスが高い濃度で検出された。動物プランクトンの個体数の割合が高い地点のサンプルほど、ノロウイルス濃度が低下していた。また、カキ中腸線のDNAメタバーコーディング解析を行い、カキが動物および植物プランクトンの双方を捕食していることも確認された。

  18. トランスクリプトーム解析による大腸菌の塩耐性獲得機構の解明

    真砂 佳史, 佐野 大輔, 久保田 健吾, 稲葉 愛美

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Challenging Exploratory Research

    2015年4月1日 ～ 2018年3月31日

    詳細を見る 詳細を閉じる

    本研究では「大腸菌は海域において塩耐性を獲得し，通常の測定法で検出されるより高濃度で広く存在している」という仮説を立て，その検証を行った。下水処理水の影響を受けている国内の沿岸海域を対象とした現地調査により，夏季に塩を添加した培地の方がコロニー数が高い傾向が認められた。しかし現地試料から単離した大腸菌株であっても，塩添加培地における増殖速度は非添加培地に比べ著しく遅く，塩の存在は生育に負の影響を与えていると考えられた。以上より，環境中の大腸菌の一部は塩耐性を持ち，条件が整えば塩存在下でも増殖できることが示された。これは，海域において大腸菌の見かけの減衰速度が遅くなることを示唆している。

  19. リアルタイム水中病原体濃度レベル予測システムの開発

    加藤 毅, 佐野 大輔, 小林 彩乃

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (C)

    研究機関：Gunma University

    2015年4月1日 ～ 2018年3月31日

    詳細を見る 詳細を閉じる

    水利用における微生物学的安全性を担保するには実時間病原性微生物予測法を実現した．水中における病原体濃度は下水を培養ベースの方法でモニタリングすることで観測されてきており，水由来の病気を防ぐのに有効だと知られている．一方，実時間モニタリングはいまだに実現していない本プロジェクトでは実時間で測定できる水質水門データから病原体を予測する方法論を確立した．その方法論は，水質工学における性質を利用して小標本からでも高精度に学習を可能にするものである．開発した方法論をオンラインソフトウェアとして実装した．

  20. 水循環システムに起因する水系感染症ウイルスの環境適応進化メカニズムの解明

    大村 達夫, 李 玉友, 佐野 大輔, 渡辺 幸三

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (A)

    研究機関：Tohoku University

    2015年4月1日 ～ 2018年3月31日

    詳細を見る 詳細を閉じる

    本研究では、ヒトノロウイルスの代替としてマウスノロウイルス、ヒトロタウイルスとしてサルロタウイルスを用い、遊離塩素への繰返し曝露がもたらすウイルス集団の適応進化に関する研究を行った。その結果、両方のウイルスに関し、遊離塩素への繰返し曝露により遊離塩素耐性集団を得ることに成功した。マウスノロウイルスに関しては外殻タンパク質遺伝子配列、サルロタウイルスに関しては全ゲノム配列の解析を行ったところ、遊離塩素への繰返し曝露により、ある特定の形質を有する株の優占が生じることが明らかとなった。以上の結果から、水処理によく用いられる遊離塩素処理が、胃腸炎ウイルスへの淘汰圧として働いていることが示された。

  21. 培養細胞のアポトーシスを指標とした下水再生水のウイルス感染リスク評価技術の開発 競争的資金

    佐野 大輔

    2016年4月 ～ 2018年3月

  22. 腸内細菌由来糖鎖が小腸上皮細胞への胃腸炎ウイルス感染に与える影響に関する研究 競争的資金

    佐野 大輔

    2015年10月 ～ 2017年9月

  23. ウイルス感染への細胞応答を活用した水中感染性腸管系ウイルスの迅速検出手法の開発

    大村 達夫, 佐野 大輔, 真砂 佳史

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Challenging Exploratory Research

    研究機関：Tohoku University

    2014年4月1日 ～ 2016年3月31日

    詳細を見る 詳細を閉じる

    本研究は、新規な感染性ウイルス検出手法に活用可能な細胞応答遺伝子を同定することを目指すものである。細胞小器官内の陽イオン濃度を制御するイオンチャネルタンパク質の遺伝子について、ウイルス感染後の発現量をモニタリングしたところ、KCNJ4遺伝子発現量は感染多重度=1で24時間後、感染多重度=0.1で36時間後に有意に増加していた。SCN7A遺伝子は、両方のMOI条件で24時間後に有意に増加していた。これらの結果から、KCNJ4およびSCN7A遺伝子が、感染性ウイルスの存在を早期に検出することが可能な遺伝子マーカーであることが示された。

  24. 水環境におけるヒトノロウイルス未知動態の解明

    大村 達夫, 原田 秀樹, 佐野 大輔, 渡部 徹, 岡部 聡, 真砂 佳史

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (A)

    研究機関：Tohoku University

    2012年4月1日 ～ 2015年3月31日

    詳細を見る 詳細を閉じる

    本研究では、水環境中におけるノロウイルスの未知動態を解明することを目的とし、１）ウイルス外殻タンパク質損傷評価法の適用条件、２）環境由来濃縮サンプルから目的ウイルスゲノムのみを特異的に回収する手法の開発、および３）ノロウイルスによる養殖カキの汚染度に関する調査・研究を行った。その結果、カプシドタンパク質損傷検出手法が水環境中のヒトノロウイルスに適用可能であることが確認され、環境由来濃縮サンプルから損傷を受けていないウイルスゲノムのみを特異的に回収する手法の確立に成功した。さらに養殖カキ中の優占している遺伝子型のみならずマイナーな感染流行株が次世代シーケンスにより検出可能であることが示された。

  25. 先進国社会における胃腸炎ウイルス適応進化による消毒剤耐性獲得メカニズムの解明 競争的資金

    佐野 大輔

    2013年4月 ～ 2015年3月

  26. 組織血液型決定抗原様物質を保持する腸内細菌がヒトノロウイルスの環境中動態に与える影響の解明 競争的資金

    佐野 大輔

    2014年4月 ～

  27. グライコミクス情報を活用した下水処理水再利用における胃腸炎ウイルス制御の新展開 競争的資金

    佐野 大輔

    2014年4月 ～

  28. 日本で発生しうる輸入感染症予見を目的としたスペインにおける下水中病原微生物の同定 競争的資金

    佐野 大輔

    2014年4月 ～

  29. 糞便汚染マーカー定量検出を基盤とした微生物学的水質管理手法の確立

    岡部 聡, 石井 聡, 佐藤 久, 佐野 大輔

    2011年4月1日 ～ 2014年3月31日

    詳細を見る 詳細を閉じる

    現行の糞便汚染指標である大腸菌群数による微生物学的水質管理は、水環境中での大腸菌群の増殖、糞便に由来しない大腸菌群の存在など、数多くの問題点が指摘されている。このため、合理的な糞便性汚染指標の確立および水域の微生物学的水質管理手法の確立が急務となっている。このような背景のもと、本研究では、宿主特異的遺伝子マーカー（腸内蛋白質分解細菌の最優占種であるBacteroides-Prevotella 属由来遺伝子）をもとに水域の糞便汚染レベルを定量的に評価し、さらに糞便汚染源（ヒト、家畜及び野生動物等）を迅速かつ正確に特定する新規方法論を確立し、具体的な汚染防止対策の構築を含む合理的な微生物学的水質管理を実現することを目的としている。 本年度は、昨年度開発した各宿主特異的糞便汚染マーカーを実際の水環境に適用し、糞便汚染の実態を明らかにした。さらに、遺伝子マーカーを糞便汚染指標として活用するためには、生存細胞と死細胞を区別して定量することが求められる。そこで、Propidium monoazide (PMA)を併用した定量PCR法を確立し、糞便汚染源の特定を行うために重要となる生菌由来の糞便性汚染マーカーの定量が可能となった。 次に、環境水中における糞便汚染マーカー（ヒト、ブタ、ウシ、ニワトリ、カモの各宿主特異的遺伝子マーカー）の挙動を解析した。さらに、既存の糞便汚染指標である大腸菌群数、糞便生大腸菌群及び大腸菌の定量も行い、減衰速度の違いを評価した。 最後に、水系感染する腸管系感染症起因細菌及びノロウイルス等の腸管系ウイルスの特異的検出を行い、各宿主特異的遺伝子マーカーと病原微生物の環境水中における存在比の相関関係を調査した。

  30. 腸内細菌由来糖鎖を活用したノロウイルスの水環境中動態解明に関する研究 競争的資金

    佐野 大輔

    2009年4月 ～ 2014年3月

  31. 組織細胞によるヒトノロウイルス培養への挑戦

    大村 達夫, 佐野 大輔, 真砂 佳史

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Challenging Exploratory Research

    研究機関：Tohoku University

    2011年 ～ 2012年

    詳細を見る 詳細を閉じる

    本プロジェクトでは、組織細胞での培養方法が確立していないヒトノロウイルスについて、組織細胞へのノロウイルスの感染確立を可能とする知見を得ることを試みた。ウイルス粒子の細胞への侵入及び脱殻過程を蛍光顕微鏡で観察する目的で、緑色蛍光タンパク質が付加したノロウイルス様中空粒子を合成することに成功したが、組織細胞への感染操作において観察に足る十分な蛍光量が確保できないことが明らかとなった。そこで、組織細胞へのノロウイルス感染が生じた際の細胞応答に着目し、ウイルス感染を迅速に検出可能な遺伝子マーカーの探索を試みた。テストウイルスとしてはポリオウイルスを用い、INT407細胞へ感染後、12時間後に総RNAを抽出し、DNAマイクロアレイを用いたトランスクリプトーム解析を行った。その結果、高い変動値を示す遺伝子としてカリウムイオンの恒常性に関わるKCNJ4が見出された。この結果は、ポリオウイルスが細胞に感染した際、細胞応答の1つとして細胞内小器官内のカリウムイオン濃度の調整が行われたことを示している。さらに、このKCNJ4遺伝子はウイルス感染以外の影響、例えば重金属の存在等によっては発現量が変動しないことも確かめられた。本研究により、ノロウイルスの細胞感染を迅速に検出するために必要な遺伝子マーカーが同定されたことから、組織細胞によるノロウイルス培養実現に向けて非常に貴重な知見が得られたと言える。

  32. 蛍光分子とウイルス吸着タンパク質を用いた革新的水中病原ウイルスセンサの開発

    佐藤 久, 佐野 大輔

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Challenging Exploratory Research

    研究機関：Hokkaido University

    2010年 ～ 2011年

    詳細を見る 詳細を閉じる

    本研究の最終目的は水中に存在する病原ウイルスを検出するセンサを開発することである。本研究期間内において、センサ開発に必須のウイルス吸着タンパク質(VBP)と蛍光分子(fluorophore)を作製した。VBPを合成するように遺伝子組み換えされた大腸菌を用いてVBPを生産した。SDS-PAGEにより生成したタンパク質の分子量がVBPの分子量である60kDaと一致することを確認した。既往の研究に従い、本研究室においてfluorophore(4, 4-difluoro-4-bora-3a, 4a-diaza-s-indacene、通称BODIPY)を生産する方法を確立した。

  33. ウイルス吸着タンパク質を用いた環境中からの病原ウイルス濃縮・検出・同定技術開発

    大村 達夫, 片山 浩之, 佐野 大輔, 渡部 徹, 真砂 佳史, 植木 洋

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (S)

    研究機関：Tohoku University

    2007年 ～ 2011年

    詳細を見る 詳細を閉じる

    本プロジェクトでは, 活性汚泥細菌が産生するウイルス吸着タンパク質(virus-binding protein : VBP)や, 加水分解酵素を用いた固形環境試料からのウイルス回収技術(enzymatic virus elution法 : EVE法)を活用し, 全く新しい病原ウイルス濃縮・検出・同定技術を開発することを目指した.その結果, 幅広い種の腸管系ウイルスを捕捉可能なVBPである腸管系ウイルス吸着タンパク質(Enteric virus-binding protein : EVBP)の分離及びクローニングに成功した他, 河川底泥・海底泥及び下水等の腸管系ウイルスの環境中動態に関わるサンプルからの効率的なウイルス回収・検出方法を確立し, さらにエチジウムモノアザイドを用いた感染性ウイルス検出技術を確立することで, 当初目的を達成することができたと言える.

  34. 先進国社会における特異的な水系感染症ウイルス進化による塩素耐性獲得メカニズム解明 競争的資金

    佐野 大輔

    2009年4月 ～ 2010年3月

  35. コミュニティーレベルでの胃腸炎ウイルス制御法開発(社会のワクチン)への挑戦

    中込 治, 中込 とよ子, 佐野 大輔

    2009年 ～ 2009年

    詳細を見る 詳細を閉じる

    ノロウイルスはどうやって現代の上下水処理制御をかいくぐり自然界でヒトからヒトへの感染環を維持しているのか。ノロウイルスによる被害を最小化するためには、ノロウイルスの自然界での動態を明らかにし、感染制御をコミュニティーレベルで達成する新規な衛生工学的水処理技術の開発が必要である。本研究は、ノロウイルスが組織血液型抗原をレセプターとすることにヒントを得て、ノロウイルスを環境中で安定に生存させるキャリア細菌があるのではないかという仮説を検証することにより、新規な下水処理法の開発につながる突破口をつかむことを目的とした。未処理下水を原材料に6種類の腸内細菌用培地を用いた微好気性培養を行い、これを血液型判定法の原理を応用したELISAによりノロウイルス吸着細菌をスクリーニングした。ここで分離された菌を微好気条件下に単離し、増菌後に組織血液型抗原への結合を確認した上で、16SrDNAの配列を決定し、ホモロジー検索によりPseudomonas,Klebsiella,Proteusに近縁な3株を同定した。これらの3株と対照菌株である大腸菌K12株とを用いて、ノロウイルス粒子の吸着実験を行ったところ、吸着平衡定数が10^12のオーダーの高い値が得られた。すなわち、菌体表面に組織血液型抗原と同等の糖鎖構造をもち、かつ高い平衡定数でノロウイルスに吸着する腸内細菌株を見出すことができた。今後、再現性および推定パラメータの精度確認などの実験が必要であるが、ノロウイルスのキャリア細菌仮説を強く支持する結果が得られたことから、コミュニティーレベルでの胃腸炎ウイルス制御法開発,すなわち,「社会のワクチン」開発に向けての第1歩が得られたと考える。

  36. ウイルス吸着タンパク質と光触媒の複合化による水中病原ウイルス除去・不活化技術開発 競争的資金

    佐野 大輔

    2004年4月 ～ 2007年3月

  37. The elucidation of inactivation mechanisms of Noroviruses with the detection of damages on viral gene and capsid protein 競争的資金

    2007年 ～

    詳細を見る 詳細を閉じる

    The investigation of norovirus, one of important causative agents for viral diseases, has a big burden that this virus can not grow with cell culture in laboratory. This means that it is impossible to evaluate the norovirus inactivation by disinfection techniques, such as chlorine treatment and ultraviolet radiation. In this study, a new method to directly detect damages on noroviral particle will be developed, which could contribute to the elucidation of the mechanism of norovirus inactivation.

  38. 水環境中の病原ウイルス除去技術開発に関する研究

    大村 達夫, 渡部 徹, 佐野 大輔

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (B)

    研究機関：Tohoku University

    2004年 ～ 2006年

    詳細を見る 詳細を閉じる

    ウイルス吸着タンパク質(Virus-Binding Protein : VBP)は研究代表者が活性汚泥中から分離することに成功した水中病原ウイルスに対する吸着親和性の高いタンパク質である.このVBPを用いた全く新しい水中病原ウイルス除去技術を開発することが本研究の目的である.平成16年度からの3年間において,以下に示す成果を得た. まず研究初年度においては,VBPの大量合成・抽出及び精製システムの構築を実現するため,大腸菌破砕法の検討及びVBP精製におけるアフィニティクロマトグラフィのスケールアップに成功した.VBPをウイルス吸着材として用いるためには大量のVBPが必要となる事から,VBP生産工程全般のスケールアップは本研究において非常に重要であった. 研究次年度では,VBPの配向固定化を実現するために,リジンタグを付加したリジンタグ付きVBP (LysTagVBP)の創出を行った.VBPの固定化の際にはタンパク質分子中のリジン残基を利用したが,その固定化位置を揃えるために,VBPのC末端にリジンが6残基連なったタグを挿入したものである.このことから,VBP固定化を効率的に行うことが可能となった. 研究最終年度においては,大量合成したLysTagVBPを固定化したウイルス除去カラムの作成及びウイルス除去能を確認する実験を行った.その結果,VBP固定化担体を用いたカラム実験では,実験開始1時間後に約40%のウイルス除去が確認され,さらに3時間後には70%のウイルス除去が確認された.VBPを利用した水中ウイルス除去技術開発は全く新しい試みであり,本研究を通じて得られたVBPの利用に関する有用な知見は,VBPを用いた水中病原ウイルス除去技術の実用化に加えてVBPに関する今後の研究の発展を促す礎になると考えられる.

  39. 豪州Moreton湾における毒素生産シアノバクテリアの増殖メカニズムの解明

    大村 達夫, 渡部 徹, 佐野 大輔

    提供機関：Japan Society for the Promotion of Science

    制度名：Grants-in-Aid for Scientific Research

    研究種目：Grant-in-Aid for Scientific Research (A)

    研究機関：Tohoku University

    2004年 ～ 2006年

    詳細を見る 詳細を閉じる

    本研究課題では,沿岸域での鉄の動態及び毒性シアノバクテリア(L.majuscula)による鉄摂取経路を解明し,沿岸域における生物利用可能な鉄の生成を予測する数値モデルを確立することを目的とした。以下に,その研究成果を示す。 沿岸域での鉄の形態及びフミン鉄の錯平衡・凝集の実態を把握するための現地調査及び室内実験を行った。その結果,河川水中においてフミン第二鉄が存在することが示され,また,フミン鉄のうち90%程度は,沿岸域でカチオンの競合により解離することが分かった。クロマトグラフィー分析を併用することで,分子量及び疎水性物質含有量が低いフミン物質ほど凝集特性は低く,沿岸域への溶存鉄輸送に大きく貢献することが明らかとなった。 沿岸域におけるフミン鉄の錯平衡モデルを,分子量やイオン強度,pH等の物理化学的要因を考慮することが可能なデバイ-ヒュッケル理論に基づき構築した。凝集モデルは,従来のDLVO理論に新たに疎水性相互作用を組み込むことで構築された。上述の錯平衡及び凝集モデルと潮流拡散モデルを融合させることで,沿岸域におけるフミン鉄濃度の空間的及び時間的分布を評価することが可能となった。 スーパーオキシドを介してL.majusculaが鉄を摂取する経路を解明し,鉄摂取機構のモデル化を行った。スーパーオキシドによるフミン鉄の生物利用性(Fe(II)の生成速度)はその起源により異なるが,酸性官能基を多く含むフミン物質に結合した鉄ほど生物利用性が低いことが明らかとなった。また,Fe(II)生成に関わる各種反応速度を測定し,L.majusclaの鉄摂取に関する新たな反応スキーム(鉄摂取モデル)が構築された。以上の研究成果より,各モデルを融合させることにより,鉄の供給形態から沿岸域での生物摂取までの一連の過程を総合的に予測・評価することが可能となった。

  40. 都市域内高度水循環システム構築にための下水処理水からの病原ウイルス除去技術開発 競争的資金

    佐野 大輔

    2002年4月 ～ 2004年3月

  41. Development of virus detection method using virus-binding protein 競争的資金

    制度名：Grant-in-Aid for Scientific Research

    2000年 ～

    詳細を見る 詳細を閉じる

    Virus-binding proteins are bacterial proteins that can exhibit the binding ability with human pathogenic viruses. It has been speculated that the presence of virus-binding proteins affects the movement of pathogenic viruses in water environment. In this study, a new method for the detection of viruses from water using the virus-binding protein as viral adsorbent is going to be developed.

  42. The elucidation of virus transmission pathway in water environment 競争的資金

    制度名：Grant-in-Aid for Scientific Research

    1997年 ～

    詳細を見る 詳細を閉じる

    It has been elucidated that some pathogenic viruses frequently causing infectious diseases among human are contaminating water environment via treated wastewater. The water environment could have an important role in the transmission of pathogenic viruses in human society, but details have remained to be clear. In this study, in order to elucidate the mechanisms of virus transmission in human society, effective virus detection methods for variety of water samples, including river water, sea water and sediments, are newly developed, and they are applied to the actual environmental water samples to know the pathway of pathogenic viruses in water environment.

︎全件表示 ︎最初の5件までを表示

担当経験のある科目(授業) 6

  1. Water and Public Health　Faculty of Engineering, Hokkaido University

  2. 工学基礎演習　北海道大学環境社会工学科

  3. 水・公衆衛生特論　北海道大学大学院工学研究院

  4. 環境工学序論　北海道大学工学部環境社会工学科

  5. 物理化学要論　北海道大学工学部環境社会工学科

  6. 分離工学　北海道大学工学部環境社会工学科

︎全件表示 ︎最初の5件までを表示