JP2021516258A - 動物における大腸菌(E.coli)に基づく感染症の予防及び/又は処置での直接給与微生物の使用 - Google Patents
動物における大腸菌(E.coli)に基づく感染症の予防及び/又は処置での直接給与微生物の使用 Download PDFInfo
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Abstract
Description
本出願は、2018年3月6日に出願された米国仮特許出願第62/639,158号明細書に対する優先権を主張し、この開示は、その全体が参照により本明細書に組み込まれる。
バチルス・スブチリス(Bacillus subtilis)3BP5株 アクセッション番号NRRL B−50510、
バチルス・アミロリケファシエンス(Bacillus amyloliqufaciens)918株 ATCCアクセッション番号NRRL B−50508、及び
バチルス・アミロリケファシエンス(Bacillus amyloliqufaciens)1013株 ATCCアクセッション番号NRRL B−50509。
1.動物における大腸菌(E.coli)に基づく感染症を予防するための及び/又は処置するための組成物であって、バチルス(Bacillus)3BP5株(NRRL B−50510);B.アミロリケファシエンス(B.amyloliquefaciens)918株(NRRL B−50508)及び15AP4株(PTA−6507)を単独で含むか又はこれらの株に由来する培養上清と組み合わせて含むバチルス属(Bacillus)ベースの直接給与微生物成分を含む組成物。
2.前記組成物は、体重増加の上昇、飼料率の増加、腸バリアの完全性の改善、死亡率の減少、及び糞便中での大腸菌(E.coli)排出の減少からなる群から選択される1種又は複数種の能力上の利点をもたらす、実施形態1に記載の組成物。
3.直接給与微生物は内生胞子の形態である、実施形態1又は2に記載の組成物。
4.前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、実施形態1又は2に記載の組成物。
5.前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、実施形態3に記載の組成物。
6.少なくとも1種の酵素は、フィターゼ、プロテアーゼ、アミラーゼ、キシラナーゼ、及びベータ−グルカナーゼからなる群から選択される、実施形態4に記載の組成物。
7.前記組成物は、飼料添加剤組成物又はプレミックスである、実施形態1、2、5、又は6のいずれか一つに記載の組成物。
8.前記組成物は、飼料添加剤組成物又はプレミックスである、実施形態3に記載の組成物。
9.前記組成物は、飼料添加剤組成物又はプレミックスである、実施形態4に記載の組成物。
10.実施形態7に記載の飼料添加剤組成物を含む飼料。
11.実施形態8又は9に記載の飼料添加剤組成物を含む飼料。
12.実施形態7に記載の飼料添加剤組成物と、投与に関する指示書とを含むキット。
13.実施形態8又は9に記載の飼料添加剤組成物と、投与に関する指示書とを含むキット。
14.動物における大腸菌(E.coli)に基づく感染症を予防する及び/又は処置する方法であって、バチルス・スブチリス(Bacillus subtilis)3BP5株(NRRL B−50510);バチルス・アミロリケファシエンス(Bacillus amyloliquefaciens)918株(NRRL B−50508)及び15AP4株(PTA−6507)を含むバチルス属(Bacillus)ベースの直接給与微生物成分を含む組成物の有効な量を投与することを含む方法。
15.組成物は、体重増加の上昇、飼料率の増加、腸バリアの完全性の改善、及び糞便中での大腸菌(E.coli)排出の減少からなる群から選択される1種又は複数種の能力上の利点をもたらす、実施形態14に記載の方法。
16.直接給与微生物は内生胞子の形態である、実施形態14又は15に記載の方法。
17.前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、実施形態14又は15に記載の方法。
18.前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、実施形態16に記載の方法。
19.少なくとも1種の酵素は、フィターゼ、プロテアーゼ、アミラーゼ、キシラナーゼ、及びベータ−グルカナーゼからなる群から選択される、実施形態17に記載の方法。
20.少なくとも1種の酵素は、フィターゼ、プロテアーゼ、アミラーゼ、キシラナーゼ、及びベータ−グルカナーゼからなる群から選択される、実施形態18に記載の方法。
21.前記組成物は、飼料添加剤組成物又はプレミックスである、実施形態14、15、19、及び20のいずれか一つに記載の方法。
22.前記組成物は、飼料添加剤組成物又はプレミックスである、実施形態16に記載の方法。
F18腸内毒素原性大腸菌(Escherichia coli)でチャレンジした離乳ブタの成長能力、糞便の大腸菌(E.coli)排出スコア、及び腸バリア機能への、バチルス属(Bacillus)の3種の株をベースとする直接給与微生物(バチルス属(Bacillus)3BP5株、918株、15AP4株)の効果
材料及び方法
飼育及び環境
動物及び実験プロトコルの使用は、Animal Experiment Committeeにより承認されている。基礎常用飼料を、給与時に、エネルギーとタンパク質とのバランスが取れているように、且つNRC(2012)により推奨されているようにこの年齢の成長期のブタの栄養素要求量を満たすか又は上回るように、配合する(表1)。
体重が6.44±0.64kgの新たに離乳した雌雄混合(去勢ブタ及び若雌ブタそれぞれ50%)ブタ合計72匹を、17日間の実験で使用する。各ブタを遺伝子検査して、F18大腸菌(E.coli)に対する感受性を確実なものにする。これらのブタを個々に秤量し、体重でブロックし、ブロック内で1〜4の常用飼料処置に無作為に割り当てる。これらのブタを、1個の檻当たり2匹のブタで収容し、これらのブタのチャレンジ状態に基づいて、下記の2つの別々の部屋の内の1つで飼育する:18匹の非チャレンジコントロ−ルブタ(9個の檻;PC処置)のためのより小さい部屋、並びに54匹のチャレンジブタ(27個の檻;NC、NC+DFM1、及びNC+DFM2に関する処置当たり9個の檻)のための他のより大きい部屋。全てのブタを、環境が管理された部屋で飼育する。各檻の片側には、ブタが飼料及び水を自由に摂取することが可能であるステンレス鋼製の自動給与器及び乳首給水器が備えられている。この実験を17日にわたり実行した。この実験は、常用飼料への7日間の適応期間と、それに続く10日間のチャレンジ期間とで構成された。離乳後7日目(この研究の7日目)に、NC処置群、NC+DFM1処置群、及びNC+DFM2処置群のブタに、F18毒素原性大腸菌(E.coli)(ETEC)を経口感染させる。PC処置群には、大腸菌(E.coli)でチャレンジしない。
体重及び飼料消費量を、0日目、7日目、及び17日目(試験終了)に測定して、平均1日増加量(ADG)、平均1日飼料摂取量(ADFI)、及び増加量対飼料比(G:F)をモニタリングする。7日目(チャレンジ直前)、並びに感染後(dpi)1日目、2日目、3日目、4日目、5日目、6日目、7日目、及び10日目に糞便スワブを採取し、大腸菌(E.coli)排出を評価する。糞便スコアを、下記の尺度を使用して、盲検方式で毎日目視にてランク付けする:1=固体;2=半固体;3=半液体;4=液体。
成長能力:7日間の適応期間の最中に、4種の処置の間に、ADG、ADFI、又はG:Fの有意差はない(P>0.10)。しかしながら、NC+DFM2による体重増加は、他の全ての処置と比べて数値的に大きい。NC+DFM2常用飼料によるブタは、この研究の最初の7日間の最中に0.781kg(最終体重−初期体重)増加し、PC、NC、及びNC+DFM1はそれぞれ、0.239、0.188、及び0.365kg増加した(表3)。
タイトジャンクション(閉塞ジャンクションとしても既知である)とは、2個の細胞の密接に関連する領域のことであり、これらの細胞の膜は互いに結合して、流体に対して実質的に不透過性のバリアを形成する。腸の健康という観点から、タイトジャンクションは、良好な腸の完全性又は不十分な腸の完全性のどちらかにつながる可能性がある腸の上皮細胞間のチャネルのことである。良好な腸の完全性が存在する場合には、クローディン3及びオクルディン等のタイトジャンクションタンパク質は、2個の上皮細胞の間で発現し、腸管内腔から体循環への病原体の転位を防止し得るバリアを提供するのに役立つ。クローディンは、タイトジャンクションタンパク質の最も重要なファミリであり、クローディン3は、これらのタイトジャンクションタンパク質をコードする遺伝子の内の1つである。オクルディンは、別の重要なタイトジャンクションタンパク質である。クローディン及びオクルディンは、同定された最初のタイトジャンクション内在性膜タンパク質であった。タイトジャンクションタンパク質RNAの測定は、タイトジャンクションが動物の消化管中で維持されていない場合には、病原体及び毒素の腸管内腔から体循環への転位を可能にし得る透過性が生じ得、そのため動物の健康を含むか、又は動物の死さえももたらすことから、バリアの完全性と腸の健康との指標として機能し得る。
Claims (23)
- 動物における大腸菌(E.coli)に基づく感染症を予防するための及び/又は処置するための組成物であって、バチルス・スブチリス(Bacillus subtilis)3BP5株(NRRL B−50510);バチルス・アミロリケファシエンス(Bacillus amyloliquefaciens)918株(NRRL B−50508)及び15AP4(PTA−6507)を単独で含むか又はこれらの株に由来する培養上清と組み合わせて含むバチルス属(Bacillus)ベースの直接給与微生物成分を含む組成物。
- 前記組成物は、体重増加の上昇、飼料率の増加、腸バリアの完全性の改善、死亡率の減少、及び糞便中での大腸菌(E.coli)排出の減少からなる群から選択される1種又は複数種の能力上の利点をもたらす、請求項1に記載の組成物。
- 前記直接給与微生物は内生胞子の形態である、請求項1又は2に記載の組成物。
- 前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、請求項1又は2に記載の組成物。
- 前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、請求項3に記載の組成物。
- 前記少なくとも1種の酵素は、フィターゼ、プロテアーゼ、アミラーゼ、キシラナーゼ、及びベータ−グルカナーゼからなる群から選択される、請求項4に記載の組成物。
- 前記組成物は、飼料添加剤組成物又はプレミックスである、請求項1、2、5、又は6のいずれか一項に記載の組成物。
- 前記組成物は、飼料添加剤組成物又はプレミックスである、請求項3に記載の組成物。
- 前記組成物は、飼料添加剤組成物又はプレミックスである、請求項4に記載の組成物。
- 請求項7に記載の飼料添加剤組成物を含む飼料。
- 請求項8又は9に記載の飼料添加剤組成物を含む飼料。
- 請求項7に記載の飼料添加剤組成物と、投与に関する指示書とを含むキット。
- 請求項8又は9に記載の飼料添加剤組成物と、投与に関する指示書とを含むキット。
- 動物における大腸菌(E.coli)に基づく感染症を予防する及び/又は処置する方法であって、バチルス・スブチリス(Bacillus subtilis)3BP5株(NRRL B−50510);バチルス・アミロリケファシエンス(Bacillus amyloliquefaciens)918(NRRL B−50508)及び15AP4(PTA−6507)を含むバチルス属(Bacillus)ベースの直接給与微生物成分を含む組成物の有効な量を投与することを含む方法。
- 前記組成物は、体重増加の上昇、飼料率の増加、腸バリアの完全性の改善、死亡率の減少、及び糞便中での大腸菌(E.coli)排出の減少からなる群から選択される1種又は複数種の能力上の利点をもたらす、請求項14に記載の方法。
- 前記直接給与微生物は内生胞子の形態である、請求項14又は15に記載の方法。
- 前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、請求項14又は15に記載の方法。
- 前記組成物は、任意選択的にカプセル化され得る少なくとも1種の酵素をさらに含む、請求項16に記載の方法。
- 前記少なくとも1種の酵素は、フィターゼ、プロテアーゼ、アミラーゼ、キシラナーゼ、及びベータ−グルカナーゼからなる群から選択される、請求項17に記載の方法。
- 前記少なくとも1種の酵素は、フィターゼ、プロテアーゼ、アミラーゼ、キシラナーゼ、及びベータ−グルカナーゼからなる群から選択される、請求項18に記載の方法。
- 前記組成物は、飼料添加剤組成物又はプレミックスである、請求項14、15、19、及び20のいずれか一項に記載の方法。
- 前記組成物は、飼料添加剤組成物又はプレミックスである、請求項16に記載の方法。
- 前記組成物は、飼料添加剤組成物又はプレミックスである、請求項17に記載の方法。
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CN113773997A (zh) * | 2021-09-30 | 2021-12-10 | 中南民族大学 | 枯草芽孢杆菌Bacillus subtilis S-2及合生制剂的配伍方法及应用 |
CN115843934A (zh) * | 2022-12-05 | 2023-03-28 | 甘肃农业大学 | 用于牦牛冷季补饲的秸秆型颗粒饲料、制备方法及应用 |
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KR20210006883A (ko) | 2021-01-19 |
US20210000892A1 (en) | 2021-01-07 |
MX2020009181A (es) | 2021-01-08 |
PH12020551413A1 (en) | 2021-09-01 |
RU2020132317A (ru) | 2022-04-06 |
TWI790356B (zh) | 2023-01-21 |
BR112020018019A2 (pt) | 2021-03-02 |
CN112368006A (zh) | 2021-02-12 |
TW201938036A (zh) | 2019-10-01 |
CA3093251A1 (en) | 2019-09-12 |
US11351207B2 (en) | 2022-06-07 |
EP3762000A1 (en) | 2021-01-13 |
WO2019173174A1 (en) | 2019-09-12 |
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