{"created":"2023-05-15T15:18:29.329615+00:00","id":4882,"links":{},"metadata":{"_buckets":{"deposit":"d2ff138d-3ec5-4f99-a7e6-d005635d6392"},"_deposit":{"created_by":12,"id":"4882","owners":[12],"pid":{"revision_id":0,"type":"depid","value":"4882"},"status":"published"},"_oai":{"id":"oai:obihiro.repo.nii.ac.jp:00004882","sets":["6:8"]},"author_link":["6812"],"item_3_alternative_title_1":{"attribute_name":"その他（別言語等）のタイトル","attribute_value_mlt":[{"subitem_alternative_title":"フィリピンにおける家畜のマダニ媒介性寄生虫病に関する分子疫学的研究","subitem_alternative_title_language":"ja"}]},"item_3_date_granted_12":{"attribute_name":"学位授与年月日","attribute_value_mlt":[{"subitem_dategranted":"2023-03-20"}]},"item_3_degree_grantor_10":{"attribute_name":"学位授与機関","attribute_value_mlt":[{"subitem_degreegrantor":[{"subitem_degreegrantor_language":"ja","subitem_degreegrantor_name":"帯広畜産大学"}],"subitem_degreegrantor_identifier":[{"subitem_degreegrantor_identifier_name":"10105","subitem_degreegrantor_identifier_scheme":"kakenhi"}]}]},"item_3_degree_name_9":{"attribute_name":"学位名","attribute_value_mlt":[{"subitem_degreename":"博士（獣医学）","subitem_degreename_language":"ja"}]},"item_3_description_11":{"attribute_name":"学位授与年度","attribute_value_mlt":[{"subitem_description":"2022","subitem_description_type":"Other"}]},"item_3_description_18":{"attribute_name":"フォーマット","attribute_value_mlt":[{"subitem_description":"application/pdf","subitem_description_type":"Other"}]},"item_3_description_7":{"attribute_name":"抄録","attribute_value_mlt":[{"subitem_description":"Babesiosis, theileriosis, and anaplasmosis, alongside other TBDs, are significant parasitic diseases that inflict adverse health effects on livestock animals. This impact directly translates to million worth of financial losses for livestock raisers globally. The use of molecular tools to uncover the epidemiology of TBDs has been instrumental in their prevention and control. Considering the economic impact, elucidating the presence of tick-borne infections in livestock using appropriate diagnostic tools is vital not only in improving herd health but also in formulating and implementing countermeasures, which are expected to help farmers to recuperate the losses from TBDs. In this dissertation, I was able to elucidate the major TBPs of horses (Chapter 1), goats (Chapter 2), and cattle (Chapter 3) in the Philippines through the application of molecular diagnostic tools. \nIn Chapter 1, the presence of equine tick-borne infections in a racehorse park from Cavite, Philippines, was evaluated. A total of 124 thoroughbred horses specifically raised for racing were sampled. Babesia caballi (12.10%; 15/124), Theileria equi (0.81%; 1/124), Anaplasma phagocytophilum (10.48%;13/124), Borrelia burgdorferi sensu lato (Bbsl) (38.71%; 48/124), A. marginale (0.81%; 1/124), and Coxiella burnetii (0.81%; 1/124) were detected in racehorses. Of the 60 positive samples, 42 were single infections and 18 were multiple infections, the most frequent of which are coinfection with Bbsl and B. caballi. Sex (p = 0.026) was found to be a significant risk factor for B. caballi infection, with female horses 5.77 times more likely to be infected with B. caballi. Sequencing analysis revealed that seven partial 18S rRNA B. caballi isolates shared 98.63–100% identity and were classified as genotype A, whereas the single T. equi sequence had 99.77% identity with GenBank isolates and was confirmed as genotype E. Eight Anaplasma 16S rRNA partial sequences were highly identical to A. phagocytophilum and A. ovis, while partial sequences of Borrelia 5–23S rRNA were most closely related to Bo. japonica and Bo. garinii-like isolates. The first molecular detection of Borrelia and Anaplasma is reported in this study, as well as the first genotyping of B. caballi and T. equi in racehorses in the Philippines. \nIn Chapter 2, a total of 396 goat samples were collected from six Philippine provinces and were molecularly screened for the presence of Babesia/Theileria and Anaplasma. A detection rate of 77.02% (305/396) and 38.64% (153/396) were noted for the respective TBPs. Six samples were positive for B. ovis (1.52%). Sex and age group were associated with higher Babesia/Theileria and Anaplasma detection rates, respectively, while significantly higher rates for both pathogens were observed in exotic goat breeds. The representative Babesia/Theileria sequences shared 89.97–97.74% identity and were most closely related to T. orientalis, T. annulata, and Theileria sp. On the other hand, Anaplasma 16S rRNA sequences were related to A. odocoilei, A. platys, and A. phagocytophilum. This is the first molecular identification of B. ovis, Theileria spp., and Anaplasma spp. in goats from the Philippines. \nIn Chapter 3, a new molecular tool was evaluated for the characterization of piroplasma species diversity in bovines. Of 162, 58.64% tested positive for piroplasma using a conventional RLB-PCR assay that targets the V4 hypervariable region of the 18S rRNA gene. The positive cattle samples were from Bohol (83.33%; 20/24), Cavite (70.97%; 44/62), and Cebu (40.79%; 31/76). By leveraging the AMPtk pipeline, the merged reads generated a total of 2,179 ASVs. The BLAST non-redundant database assigned the taxonomy of 175 ASVs, which were then dereplicated into 97 ASVs. Further filtering yielded a final count of 79 distinct ASVs. The taxonomy hits of the 79 ASVs corresponding to 10 species were B. bovis (n = 37), B. bigemina (n = 18), T. orientalis (n = 13), Babesia sp. (n = 3), Hepatozoon canis (n = 2), Sarcocystis cruzi (n = 2), T. annulata (n = 1), T. equi (n = 1), T. mutans (n = 1), and Theileria sp. Thung Song (n = 1). Based on the phylogenetic analysis, ASVs assigned to B. bovis and Babesia sp. showed three major subclades and were 90.76–100% identical with B. bovis isolates deposited in GenBank. On the other hand, the evolutionary inference suggested that most of the B. bigemina and Babesia sp. ASVs were phylogenetically distinct from previously reported isolates, as evidenced by the strong statistical support, and shared 90.23–99.77% identity with isolates from the database. Thirteen T. orientalis ASVs (91.72–100% identical to other T. orientalis isolates) clustered in the T. orientalis complex subclade, 4 of which formed a subgroup with high nodal support. As anticipated, ASVs taxonomically recognized as T. annulata, T. equi, and T. mutans were separated into groups based on the species to which they belong. Furthermore, the Hepatozoon ASVs were distinctly divergent from previously detected Philippine canine H. canis isolates, while phylogenetic analysis revealed that Sarcocystis ASVs were closely related to bovine S. cruzi isolates from Malaysia and India. \nThe fundamental purpose of this research was to map out the different TBPs infecting economically important livestock in the Philippines. The presented findings from the three chapters of this dissertation attest to the attainment of this purpose. The circulating TBP populations and the extent of tick-borne infections in horses, goats, and cattle in the Philippines were revealed through the use of molecular detection tools, proving the applicability of these platforms in identifying tick-borne infections in different livestock. These results are vital in ascertaining the TBD status in the Philippines, where infections are widely present but often overlooked and neglected. The findings from these studies shall be beneficial in crafting and implementing livestock tick and TBD control and prevention programs in the country.","subitem_description_language":"en","subitem_description_type":"Abstract"},{"subitem_description":"バベシア症、タイレリア症、アナプラズマ症は、他のマダニ媒介感染症（TBD）と並んで、家畜の健康に悪影響を与える重要な寄生虫病である。こんら影響は、世界中の畜産農家が被る甚大な経済的損失に直結する。このような経済的損失を低減するために、適切な分子診断手法を用いて家畜のマダニ媒介性感染症の流行実態の一端を明らかにすることは、家畜の健康状態を改善するだけでなく、対策の立案・実施に不可欠である。そこで、本研究ではフィリピンにおけるウマ（第1章）、ヤギ（第2章）、およびウシ（第3章）の主要なマダニ媒介性病原体（TBP）の分子疫学調査を実施した。 \n第1章では、フィリピンカビテ州の競走馬牧場におけるマダニ媒介性感染症について調べた。サラブレッド馬124頭から血液サンプルを採集し、各種寄生虫の核酸検出を行ったところ、Babesia caballi（12.10％；15/124）、Theileria equi（0.81％；1/124）、Anaplasma phagocytophilum（10.48％；13/124）、Borrelia burgdorferi sensu lato（Bbsl）（38.71％；48/124）、A. marginale（0.81％；1/124）、Coxiella burnetii（0.81％；1/124)が検出された。陽性60検体のうち、42検体が単独感染，18検体が混合感染であり、BbslとB. caballiとの重複感染が最も多かった。B. caballiとT. equiについて、遺伝子型を調べたところ、それぞれA型とE型に分類された。これらの結果のなかで、BorreliaとAnaplasmaの検出とB. caballiとT. equiの遺伝子型の特定は、いずれもフィリピンのウマにおいては初の報告である。 \n第2章では、フィリピン6州から計396頭のヤギ血液サンプルを採集し、マダニ媒介性病原体の核酸検査を行った。Babesia/Theileria属およびAnaplasma属の病原体の検出率はそれぞれ77.02%（305/396）、38.64%（153/396）であった。Babesia/Theileria属およびAnaplasma属の検出率は、それぞれ外来種と高年齢層で高かった。Babesia/Theileria の代表的な塩基配列は89.97-97.74%の同一性を持ち、T. orientalis、T. annulata、Theileria sp.に最も近縁であった。B. ovisは6検体（1.52％）で陽性であった。一方、Anaplasma 16S rRNA 塩基配列はA. odocoilei、A. platys、A. phagocytophilumに近縁であった。これらの結果のなかで、B. ovis、 Theileria spp.およびAnaplasma spp.は、フィリピンのヤギにおける初の知見である。 \n第3章では、ウシにおけるピロプラズマの種多様性を特徴付けるための新しい分子ツールを評価した。18S rRNA遺伝子のV4超可変領域を標的とした従来のRLB-PCR法で162頭中58.64%がピロプラズマ（Babesia属/Theileria属）陽性と判定された。陽性ウシは、ボホール（ 83.33%; 20/24）、カビテ（ 70.97%; 44/62）、セブ（ 40.79%; 31/76）由来であった。AMPtkパイプラインを用いると、マージされたリードから合計2,179個のASVが生成された。BLAST non-redundantデータベースにより175個のASVが分類され、さらに97個のASVに分割された。フィルタリングを行った結果、最終的に79個の異なるASVを数えることができた。79個のASVのうち10種に対応する分類のヒットは、B. bovis (n = 37) 、 B. bigemina (n = 18) 、 T. orientalis (n = 13) 、 Babesia sp. (n = 3）、Hepatozoon canis（n = 2）、Sarcocystis cruzi（n = 2）、T. annulata（n = 1）、T. equi（n = 1）、T. mutans（n = 1）およびTheileria sp. Thung Song（n = 1）であった。 系統解析の結果、B. bovisおよびBabesia sp.に割り当てられたASVは3つの主要なサブクレードを示し、GenBankに登録されているB. bovis分離株と90.76～100％の同一性を有していた。 一方、進化推論では、B. bigeminaとBabesia sp.のASVの多くは、既報告分離株とは系統的に異なることが示唆された。 13種のT. orientalis ASV（他のT. orientalis分離株と91.72〜100%同一）はT. orientalis complexサブクレードにクラスター化し、そのうち4種は高いノードサポートを持つサブグループを形成していた。予想通り、分類学的にT. annulata、T. equi、T. mutansと認識されているASVは、その属する種に基づきグループに分けられた。さらに、Hepatozoon ASVsは、これまでに検出されたフィリピン由来イヌのH. canis分離株とは明確に分岐しており、Sarcocystis ASVsはマレーシアおよびインド産由来のS. cruzi分離株に近縁であることが明らかとなった。 \n本研究の総括的な目的は、フィリピンにおける経済的に重要な家畜に感染している様々なマダニ媒介性病原体を特定することであった。特に第3章で得られた知見は、この目的の達成を証明するものである。フィリピンのウマ、ヤギ、およびウシにおけるマダニ媒介性病原体の流行と分布が分子検出ツールによって明らかになり、異なる家畜におけるマダニ媒介性感染症の特定にこれらのプラットフォームが適用可能であることが証明された。これらの結果はフィリピンの家畜でこれまでに見過ごされがちであったマダニ媒介感染症の状況を把握する上で極めて重要である。これらの研究結果は、今後フィリピンにおける家畜のマダニ媒介性感染症の管理・予防プログラムの策定・実施に有益なものとなるであろう。","subitem_description_language":"ja","subitem_description_type":"Abstract"}]},"item_3_description_8":{"attribute_name":"内容記述","attribute_value_mlt":[{"subitem_description":"博士学位論文\n大学院畜産学研究科 獣医学専攻\nDoctoral Program of Veterinary Science","subitem_description_language":"ja","subitem_description_type":"Other"}]},"item_3_dissertation_number_13":{"attribute_name":"学位授与番号","attribute_value_mlt":[{"subitem_dissertationnumber":"甲第128号"}]},"item_3_publisher_5":{"attribute_name":"公開者","attribute_value_mlt":[{"subitem_publisher":"帯広畜産大学","subitem_publisher_language":"ja"}]},"item_3_rights_14":{"attribute_name":"権利","attribute_value_mlt":[{"subitem_rights":"GALON Eloiza May Saldua","subitem_rights_language":"en"}]},"item_3_subject_20":{"attribute_name":"日本十進分類法","attribute_value_mlt":[{"subitem_subject":"649","subitem_subject_scheme":"NDC"}]},"item_3_version_type_19":{"attribute_name":"著者版フラグ","attribute_value_mlt":[{"subitem_version_resource":"http://purl.org/coar/version/c_970fb48d4fbd8a85","subitem_version_type":"VoR"}]},"item_access_right":{"attribute_name":"アクセス権","attribute_value_mlt":[{"subitem_access_right":"open access","subitem_access_right_uri":"http://purl.org/coar/access_right/c_abf2"}]},"item_creator":{"attribute_name":"著者","attribute_type":"creator","attribute_value_mlt":[{"creatorAffiliations":[{"affiliationNameIdentifiers":[{"affiliationNameIdentifierScheme":"ISNI","affiliationNameIdentifierURI":"http://www.isni.org/isni/"}],"affiliationNames":[{"affiliationNameLang":"ja"}]}],"creatorNames":[{"creatorName":"GALON, Eloiza May Saldua","creatorNameLang":"en"}],"familyNames":[{}],"givenNames":[{}],"nameIdentifiers":[{}]}]},"item_files":{"attribute_name":"ファイル情報","attribute_type":"file","attribute_value_mlt":[{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2023-04-21"}],"displaytype":"detail","filename":"Galon.pdf","filesize":[{"value":"4.4 MB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"全文　Full text","objectType":"fulltext","url":"https://obihiro.repo.nii.ac.jp/record/4882/files/Galon.pdf"},"version_id":"00e24f9f-79c7-4248-a675-2bfa48ca10e1"},{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2023-04-21"}],"displaytype":"detail","filename":"Abstract(Galon).pdf","filesize":[{"value":"183.0 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"全文の要旨　Abstract","url":"https://obihiro.repo.nii.ac.jp/record/4882/files/Abstract(Galon).pdf"},"version_id":"15a49fbb-07b7-4576-a810-c5b8c546683c"},{"accessrole":"open_date","date":[{"dateType":"Available","dateValue":"2023-04-21"}],"displaytype":"detail","filename":"Evaluation(Galon).pdf","filesize":[{"value":"237.6 kB"}],"format":"application/pdf","licensetype":"license_note","mimetype":"application/pdf","url":{"label":"審査の要旨　Evaluation","url":"https://obihiro.repo.nii.ac.jp/record/4882/files/Evaluation(Galon).pdf"},"version_id":"4b1cba76-c665-4971-b395-ced5ab162946"}]},"item_language":{"attribute_name":"言語","attribute_value_mlt":[{"subitem_language":"eng"}]},"item_resource_type":{"attribute_name":"資源タイプ","attribute_value_mlt":[{"resourcetype":"doctoral thesis","resourceuri":"http://purl.org/coar/resource_type/c_db06"}]},"item_title":"Molecular epidemiological studies on livestock tick-borne parasitic diseases in the Philippines","item_titles":{"attribute_name":"タイトル","attribute_value_mlt":[{"subitem_title":"Molecular epidemiological studies on livestock tick-borne parasitic diseases in the Philippines","subitem_title_language":"en"}]},"item_type_id":"3","owner":"12","path":["8"],"pubdate":{"attribute_name":"PubDate","attribute_value":"2023-04-21"},"publish_date":"2023-04-21","publish_status":"0","recid":"4882","relation_version_is_last":true,"title":["Molecular epidemiological studies on livestock tick-borne parasitic diseases in the Philippines"],"weko_creator_id":"12","weko_shared_id":-1},"updated":"2024-01-18T08:14:28.172002+00:00"}