JP6266831B1 - Lactic acid bacteria and feed / fertilizer / viable bacteria containing the same - Google Patents
Lactic acid bacteria and feed / fertilizer / viable bacteria containing the same Download PDFInfo
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- JP6266831B1 JP6266831B1 JP2017128717A JP2017128717A JP6266831B1 JP 6266831 B1 JP6266831 B1 JP 6266831B1 JP 2017128717 A JP2017128717 A JP 2017128717A JP 2017128717 A JP2017128717 A JP 2017128717A JP 6266831 B1 JP6266831 B1 JP 6266831B1
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 154
- 241000894006 Bacteria Species 0.000 title claims abstract description 97
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 77
- 239000004310 lactic acid Substances 0.000 title claims abstract description 77
- 239000003337 fertilizer Substances 0.000 title claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 9
- 239000003795 chemical substances by application Substances 0.000 abstract description 7
- 239000002689 soil Substances 0.000 abstract description 6
- 241001465754 Metazoa Species 0.000 abstract description 5
- 241000233866 Fungi Species 0.000 abstract description 2
- 238000012360 testing method Methods 0.000 description 14
- 235000013601 eggs Nutrition 0.000 description 9
- 108010000912 Egg Proteins Proteins 0.000 description 8
- 102000002322 Egg Proteins Human genes 0.000 description 8
- 239000013535 sea water Substances 0.000 description 8
- 241000287828 Gallus gallus Species 0.000 description 7
- 235000013330 chicken meat Nutrition 0.000 description 7
- 241000227653 Lycopersicon Species 0.000 description 6
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 6
- 239000002773 nucleotide Substances 0.000 description 6
- 125000003729 nucleotide group Chemical group 0.000 description 6
- 102000016938 Catalase Human genes 0.000 description 5
- 108010053835 Catalase Proteins 0.000 description 5
- 238000012300 Sequence Analysis Methods 0.000 description 5
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 4
- 241000186660 Lactobacillus Species 0.000 description 4
- 240000006024 Lactobacillus plantarum Species 0.000 description 4
- 235000014103 egg white Nutrition 0.000 description 4
- 210000000969 egg white Anatomy 0.000 description 4
- 210000002969 egg yolk Anatomy 0.000 description 4
- 235000013345 egg yolk Nutrition 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 238000013095 identification testing Methods 0.000 description 4
- 229940039696 lactobacillus Drugs 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 241000194033 Enterococcus Species 0.000 description 3
- 238000003794 Gram staining Methods 0.000 description 3
- 240000001929 Lactobacillus brevis Species 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 3
- 241001228709 Suruga Species 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 239000003208 petroleum Substances 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 241000251468 Actinopterygii Species 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241001522957 Enterococcus casseliflavus Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 241000186684 Lactobacillus pentosus Species 0.000 description 2
- 241000186610 Lactobacillus sp. Species 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 235000013618 yogurt Nutrition 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 235000013957 Lactobacillus brevis Nutrition 0.000 description 1
- 108010053229 Lysyl endopeptidase Proteins 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- LFYJSSARVMHQJB-QIXNEVBVSA-N bakuchiol Chemical compound CC(C)=CCC[C@@](C)(C=C)\C=C\C1=CC=C(O)C=C1 LFYJSSARVMHQJB-QIXNEVBVSA-N 0.000 description 1
- 238000010876 biochemical test Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 238000006395 oxidase reaction Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000013081 phylogenetic analysis Methods 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Landscapes
- Fodder In General (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Fertilizers (AREA)
Abstract
【課題】飼料や肥料や生菌剤などに適用することが可能な乳酸菌を提供すること。【解決手段】本発明では、受託番号：ＮＩＴＥ Ｐ−０２４９０又は受託番号：ＮＩＴＥ Ｐ−０２４９１若しくは受託番号：ＮＩＴＥ Ｐ−０２４９２で寄託されている乳酸菌を提供するものである。また、本発明では、これらの乳酸菌を含有する飼料や肥料や生菌剤を提供するものである。これらの乳酸菌は、人間を含む動物や植物などの内部で死滅することなく増殖することが確認された。また、これらの乳酸菌は、水や土などの内部で死滅することなく増殖することが確認された。そのため、これらの乳酸菌は、飼料や肥料や生菌剤などとして幅広く利用することができるものである。【選択図】なしAn object of the present invention is to provide a lactic acid bacterium that can be applied to feeds, fertilizers, viable fungi and the like. The present invention provides a lactic acid bacterium that has been deposited under the deposit number: NITE P-02490 or the deposit number: NITE P-02491 or the deposit number: NITE P-02492. Moreover, in this invention, the feed, fertilizer, and viable agent which contain these lactic acid bacteria are provided. It was confirmed that these lactic acid bacteria proliferate without being killed inside animals and plants including humans. In addition, it was confirmed that these lactic acid bacteria proliferate without being killed inside water or soil. Therefore, these lactic acid bacteria can be widely used as feeds, fertilizers, viable bacteria agents, and the like. [Selection figure] None
Description
本発明は、乳酸菌及び同乳酸菌を含有する飼料・肥料・生菌剤に関するものである。 The present invention relates to a lactic acid bacterium and a feed, a fertilizer, and a live bacterium containing the lactic acid bacterium.
乳酸菌は、代謝により乳酸を産生する細菌類の総称であり、それぞれの細菌によって異なる性質を有している。 Lactic acid bacteria are a general term for bacteria that produce lactic acid by metabolism, and have different properties depending on each bacterium.
そのため、従来より、特性の細菌（乳酸菌）を抽出し、その性質を研究し、その性質を利用した薬剤や飲食料品などの商品の開発が行われている（たとえば、特許文献１参照。）。 Therefore, conventionally, characteristic bacteria (lactic acid bacteria) are extracted, their properties are studied, and products such as drugs and foods and drinks using the properties have been developed (for example, see Patent Document 1). .
従来知られている乳酸菌は、一般的に環境によって増殖が抑制されてしまうことから、飼料や肥料や生菌剤などには適さないことが多かった。 Conventionally known lactic acid bacteria are generally unsuitable for feeds, fertilizers, viable bacteria agents and the like because their growth is generally suppressed by the environment.
そこで、本発明者は、乳酸菌の抽出、研究、検討を重ねたところ、本願発明に係る乳酸菌を見出すに至った。 Therefore, the present inventor has repeatedly extracted, researched and studied lactic acid bacteria, and has found the lactic acid bacteria according to the present invention.
請求項１に係る本発明では、受託番号：ＮＩＴＥ Ｐ−０２４９０で寄託されている乳酸菌を提供する。 The present invention according to claim 1 provides the lactic acid bacteria deposited under the deposit number: NITE P-02490.
また、請求項２に係る本発明では、受託番号：ＮＩＴＥ Ｐ−０２４９１で寄託されている乳酸菌を提供する。 Moreover, in this invention which concerns on Claim 2, the lactic acid bacteria deposited by accession number: NITE P-02491 are provided.
また、請求項３に係る本発明では、受託番号：ＮＩＴＥ Ｐ−０２４９２で寄託されている乳酸菌を提供する。 Moreover, in this invention which concerns on Claim 3, the lactic acid bacteria deposited by accession number: NITE P-02492 are provided.
また、請求項４に係る本発明では、請求項１〜請求項３のいずれかに記載の乳酸菌を含有する飼料を提供する。 Moreover, in this invention which concerns on Claim 4, the feed containing the lactic acid bacteria in any one of Claims 1-3 is provided.
また、請求項５に係る本発明では、請求項１〜請求項３のいずれかに記載の乳酸菌を含有する肥料を提供する。 Moreover, in this invention which concerns on Claim 5, the fertilizer containing the lactic acid bacteria in any one of Claims 1-3 is provided.
また、請求項６に係る本発明では、請求項１〜請求項３のいずれかに記載の乳酸菌を含有する生菌剤を提供する。 Moreover, in this invention which concerns on Claim 6, the probiotic agent containing the lactic acid bacteria in any one of Claims 1-3 is provided.
本発明に係る乳酸菌は、飼料や肥料や生菌剤や飲食物などに有効に利用することができる。 The lactic acid bacteria according to the present invention can be effectively used for feeds, fertilizers, live bacteria agents, foods and drinks, and the like.
魚を特殊な方法で加工処理して発酵させたところ、魚肉等に多種の細菌が含有されていることが検出された。その細菌の中には乳酸菌が含有されていることが検出された。 When the fish was processed and fermented by a special method, it was detected that various kinds of bacteria were contained in the fish meat. It was detected that the bacteria contained lactic acid bacteria.
そこで、乳酸菌だけを分離することにした。乳酸菌の分離には、株式会社テクノスルガ・ラボ（静岡県静岡市清水区長崎３３０番地）に細菌分離を依頼した。 Therefore, it was decided to isolate only lactic acid bacteria. For separation of lactic acid bacteria, we requested Techno Suruga Lab Co., Ltd. (330 Nagasaki, Shimizu-ku, Shizuoka City, Shizuoka Prefecture) for bacterial separation.
乳酸菌の分離は、以下の条件で分離培養し、コロニー観察を行い、グラム染色及びカタラーゼ試験を行った。
・培養条件
培地：ＭＲＳ寒天（Oxoid,Hampshire,UK） ＋３％ Nacl
培養温度：３０℃
培養期間：３日
希釈液：生理食塩水
希釈倍率：原液〜104
分離方法：希釈平板法（10μl表面塗抹）
その他条件：嫌気
・コロニー観察
実体顕微鏡SZH10（オリンパス、東京）を使用
・グラム染色
グラム染色としてファイバーＧ「ニッスイ」（日水製薬、東京）を、
顕微鏡として光学顕微鏡BX51（オリンパス、東京）を使用
・カタラーゼ試験
３％過酸化水素水を用い、気泡が発生するものを陽性と判定
Lactic acid bacteria were separated and cultured under the following conditions, colony observation was performed, and Gram staining and catalase test were performed.
Culture conditions Medium: MRS agar (Oxoid, Hampshire, UK) + 3% Nacl
Culture temperature: 30 ° C
Culture period: 3 days Diluent: physiological saline Dilution ratio: Stock solution ~ 10 4
Separation method: Dilution plate method (10μl surface smear)
Other conditions: Anaerobic / colony observation Using stereo microscope SZH10 (Olympus, Tokyo) Gram staining Fiber G “Nissui” (Nissui Pharmaceutical, Tokyo) as Gram staining,
Optical microscope BX51 (Olympus, Tokyo) is used as a microscope. Catalase test Using 3% hydrogen peroxide solution, a bubble is judged positive.
分離培養を行ったところ、性状の異なる複数のコロニーの生育が認められた。分離された各菌株について、グラム染色及びカタラーゼ試験を行ったところ、グラム陽性・カタラーゼ陰性の細菌を乳酸菌として判定し、その結果、３種類の乳酸菌と考えられるコロニーが分離された。ここでは、それぞれをSIID21336-L1、SIID21336-L2、SIID21336-L3とする。 When separate culture was performed, growth of a plurality of colonies having different properties was observed. About each isolate | separated strain, when Gram dyeing | staining and the catalase test were done, the bacteria of Gram positive and catalase negative were determined as lactic acid bacteria, As a result, the colonies considered as three types of lactic acid bacteria were isolate | separated. Here, these are assumed to be SIID21336-L1, SIID21336-L2, and SIID21336-L3, respectively.
次に、それぞれの菌（SIID21336-L1、SIID21336-L2、SIID21336-L3）について同定試験を行った。同定試験には、株式会社テクノスルガ・ラボに同定試験（細菌Premium）を依頼した。 Next, an identification test was performed for each of the bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3). For the identification test, we requested an identification test (Bacteria Premium) from Techno Suruga Lab.
同定試験は、以下の条件で培養し、16SrDNA（16SrRNA遺伝子）塩基配列解析、形態観察及び生理・生化学試験（細菌第一段階試験及び細菌第二段階試験）の結果から、帰属分類群を推定した。 In the identification test, culture is performed under the following conditions, and the attribution taxon is estimated from the results of 16SrDNA (16SrRNA gene) nucleotide sequence analysis, morphology observation and physiological / biochemical tests (bacteria first-stage test and bacteria second-stage test) did.
・培養条件
培地：ＭＲＳ寒天（Oxoid,Hampshire,UK）
培養温度：３０℃
培養期間：４８時間
希釈液：生理食塩水
その他条件：嫌気
・16SrDNA（16SrRNA遺伝子）塩基配列解析
PCR増幅からサイクルシークエンスまでの操作は各プロトコールに基づいた。
DNA抽出：アクロモペプチダーゼ（和光純薬、大阪）
PCR増幅：PrimeSTAR HS DNA Polymerase（タカラバイオ、滋賀）
サイクルシークエンス：BigDye Terminator v3.1 Cycle Sequencing Kit
（Applied Biosystems,CA,USA)
使用プライマー：PCR増幅；9F,1510R
シークエンス；9F,785F,802R,1510R
シークエンス：ABI PRISM 3130xlGenetic Analyzer System
（Applied Biosystems,CA,USA)
塩基配列決定：ChromasPro1.7（Technelysium Pty Ltd.,Tewantin,AUS)
BLAST相同性検索及び簡易分子系統解析：微生物同定用DNAデータベースDB0BA10.0
（テクノスルガ・ラボ、静岡）
国際塩基配列データベース
（GenBank/DDBJ/EMBL)
・細菌第一段階試験
光学顕微鏡（BX50F4（オリンパス、東京））による形態観察及びBARROWらの方法（BARROW,(G.I.) and FELTHAM,(R.K.A.):Cowan and Steel's Manual for the Identification of Medical Bacteria.3rd edition.1993,Cambridge University Press.)に基づき、カタラーゼ反応、オキシダーゼ反応、ブドウ糖からの酸／ガス産生、ブドウ糖の酸化／発酵について試験を行った。
・細菌第二段階試験
キット（API20Step(bioMerieux,Lyon,France)を用いた。また、英国NCIMB Ltd.（http://www.ncimb.co.uk/)との技術提携事項及び分類・同定の関連文献に従い、追加試験を実施した。
・ Culture conditions Medium: MRS agar (Oxoid, Hampshire, UK)
Culture temperature: 30 ° C
Culture period: 48 hours Diluent: physiological saline Other conditions: Anaerobic 16SrDNA (16SrRNA gene) nucleotide sequence analysis
The procedure from PCR amplification to cycle sequencing was based on each protocol.
DNA extraction: Achromopeptidase (Wako Pure Chemicals, Osaka)
PCR amplification: PrimeSTAR HS DNA Polymerase (Takara Bio, Shiga)
Cycle sequence: BigDye Terminator v3.1 Cycle Sequencing Kit
(Applied Biosystems, CA, USA)
Primer used: PCR amplification; 9F, 1510R
Sequence: 9F, 785F, 802R, 1510R
Sequence: ABI PRISM 3130xlGenetic Analyzer System
(Applied Biosystems, CA, USA)
Base sequence determination: ChromasPro1.7 (Technelysium Pty Ltd., Tewantin, AUS)
BLAST homology search and simple molecular phylogenetic analysis: DNA database DB0BA10.0 for microorganism identification
(Techno Suruga Lab, Shizuoka)
International nucleotide sequence database
(GenBank / DDBJ / EMBL)
・ Bacteria first stage test Morphological observation by optical microscope (BX50F4 (Olympus, Tokyo)) and method of BARROW et al. (BARROW, (GI) and FELTHAM, (RKA): Cowan and Steel's Manual for the Identification of Medical Bacteria. 3rd edition 1993, Cambridge University Press.) Catalase reaction, oxidase reaction, acid / gas production from glucose, oxidation / fermentation of glucose were tested.
-Bacteria second stage test kit (API20Step (bioMerieux, Lyon, France) was used. Additional tests were performed according to relevant literature.
SIID21336-L1は、16SrDNA塩基配列解析の結果、Lactobacillus属のL.brevisに帰属する可能性が示唆された。 SIID21336-L1 was suggested to belong to L. brevis of the genus Lactobacillus as a result of 16S rDNA nucleotide sequence analysis.
また、SIID21336-L1は、細菌第一段階試験の結果、Lactobacillus属の性状と一致し、細菌第二段階試験の結果、L.brevisの性状と一致した。 In addition, SIID21336-L1 was consistent with the properties of Lactobacillus as a result of the bacterial first stage test, and was consistent with the properties of L. brevis as a result of the bacterial second stage test.
以上のことから、SIID21336-L1は、Lactobacillus brevisに帰属するものと推定された。 Based on the above, it was estimated that SIID21336-L1 belongs to Lactobacillus brevis.
また、SIID21336-L2は、16SrDNA塩基配列解析の結果、Lactobacillus属のL.pentosus又はL.plantarumに近縁なLactobacillus sp.に帰属する可能性が示唆された。 Further, as a result of 16S rDNA nucleotide sequence analysis, SIID21336-L2 was suggested to be attributed to Lactobacillus sp., Which is closely related to Lactobacillus genus L.pentosus or L.plantarum.
また、SIID21336-L2は、細菌第一段階試験の結果、Lactobacillus属の性状と一致し、細菌第二段階試験の結果、L.pentosusの性状とは異なり、L.plantarumの２亜種のうち、L.plantarum subsp.argentoratensisの性状と異なり、L.plantarum subsp.plantarumの性状と一致した。 Moreover, SIID21336-L2 is consistent with the characteristics of the genus Lactobacillus as a result of the first stage test of bacteria, and as a result of the second stage test of bacteria, the characteristics of L.pentosus are different, Unlike the properties of L. plantarum subsp. Argentoratensis, it was consistent with the properties of L. plantarum subsp. Plantarum.
以上のことから、SIID21336-L2は、L.plantarumに近縁なLactobacillus sp.に帰属するものと推定された。 Based on the above, it was estimated that SIID21336-L2 belongs to Lactobacillus sp., Which is closely related to L. plantarum.
また、SIID21336−L3は、16SrDNA塩基配列解析の結果、Enterococcus属のE.casseliflavusに帰属する可能性が示唆された。 In addition, SIID21336-L3 was suggested to belong to E. casseliflavus of the genus Enterococcus as a result of 16S rDNA nucleotide sequence analysis.
また、SIID21336-L3は、細菌第一段階試験の結果、、Enterococcus属の性状と一致し、細菌第二段階試験の結果、E.casseliflavusの性状と一致した。 Moreover, SIID21336-L3 was consistent with the properties of the genus Enterococcus as a result of the bacterial first stage test, and was consistent with the properties of E. casseliflavus as a result of the bacterial second stage test.
以上のことから、SIID21336−L3は、Enterococcus.casseliflavusに帰属するものと推定された。 From the above, it was estimated that SIID21336-L3 belongs to Enterococcus.casseliflavus.
これらの乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3）は、下記のように寄託している。
寄託機関：独立行政法人製品評価技術基盤機構（NITE）特許微生物寄託センター（NPMD)（日本国千葉県木更津市かずさ鎌足２−５−８ 郵便番号292-0818）
受託日：２０１７年６月１２日
受託番号：SIID21336-L1について、ＮＩＴＥ Ｐ−０２４９０
SIID21336-L2について、ＮＩＴＥ Ｐ−０２４９１
SIID21336-L3について、ＮＩＴＥ Ｐ−０２４９２
These lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3) are deposited as follows.
Depositary institution: National Institute of Technology and Evaluation (NITE) Patent Microorganism Depositary Center (NPMD) (2-5-8 Kazusa Kamashi, Kisarazu City, Chiba, Japan Postal Code 292-0818)
Acceptance date: June 12, 2017 Accession number: NITE P-02490 for SIID21336-L1
About SIID21336-L2, NITE P-02491
About SIID21336-L3, NITE P-02492
乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3）には、以下に記載する性質を有することが確認された。なお、以下において乳酸菌の検出は、一般財団法人日本食品分析センター（東京都渋谷区元代々木町52番１号）又は一般財団法人佐賀県環境科学検査協会（佐賀県佐賀市光１丁目４番２号）において行った。 Lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3) were confirmed to have the properties described below. In the following, lactic acid bacteria are detected by the Japan Food Analysis Center (52-1 Motoyoyogi-cho, Shibuya-ku, Tokyo) or the Saga Prefectural Environmental Science Inspection Association (1-2-2 Hikari, Saga City, Saga Prefecture). No.).
（１）飼料
乳酸菌が含有されていない飼料をニワトリに与え、そのニワトリが産んだ卵を調べたところ、卵（卵黄及び卵白）から乳酸菌が検出されなかった。
(1) Feed When a feed containing no lactic acid bacteria was given to chickens and the eggs produced by the chickens were examined, no lactic acid bacteria were detected from the eggs (egg yolk and egg white).
また、乳酸菌が含有されていない上記飼料に市販されている代表的な乳酸菌（ヨーグルト）を含有させたものを飼料としてニワトリに与え、そのニワトリが産んだ卵を調べたところ、卵（卵黄及び卵白）から乳酸菌が検出されなかった。 In addition, when a chicken containing a typical lactic acid bacterium (yogurt) which is commercially available in the above feed containing no lactic acid bacteria was given to the chicken and the eggs produced by the chicken were examined, eggs (egg yolk and egg white) were examined. ) No lactic acid bacteria were detected.
しかしながら、乳酸菌が含有されていない上記飼料に上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）を含有させたものを飼料としてニワトリに与え、そのニワトリが産んだ卵を調べたところ、卵（卵黄及び卵白）から乳酸菌が検出された。 However, the feed containing no lactic acid bacteria containing the lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three) is fed to chickens as feed. When eggs laid by the chicken were examined, lactic acid bacteria were detected from the eggs (egg yolk and egg white).
このことから、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3）は、一般的な乳酸菌とは異なり、飼料に含有させることで、母体内で消化や死滅などすることなく増殖して卵の卵黄及び卵白にまで含有される性質であることが明らかとなった。 Therefore, unlike the general lactic acid bacteria, the above lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3) can be contained in the feed, and can be proliferated without being digested or killed in the mother body. It became clear that it was a property contained in egg yolk and egg white.
このように、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3）は、飼料に含有させることで、家畜等の動物の体内で増殖し、飼料を食した動物に乳酸菌を投与することができるとともに、その動物の子や卵（飼料を直接食していない子や卵）にまで含有させることができる。なお、これは、乳酸菌を含有する卵の製造方法としても利用できる。 As described above, the lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3) can be contained in the feed so that they grow in animals such as livestock, and the lactic acid bacteria can be administered to animals that have eaten the feed. In addition, it can be contained even in the pups and eggs of the animals (the pups and eggs that do not eat the feed directly). This can also be used as a method for producing eggs containing lactic acid bacteria.
（２）肥料
乳酸菌が含有されていない肥料をトマトの苗に与え、成熟したトマトを調べたところ、トマトから乳酸菌が検出されなかった。
(2) Fertilizer Fertilizer containing no lactic acid bacteria was given to tomato seedlings, and mature tomatoes were examined. No lactic acid bacteria were detected from the tomatoes.
しかしながら、乳酸菌が含有されていない上記肥料に上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）を含有させたものを肥料としてトマトの苗に与え、成熟したトマトを調べたところ、トマトから乳酸菌が検出された。また、乳酸菌が含有されていない肥料を与えた場合よりも良好に（早く、多く）成熟することが確認された。 However, the fertilizer containing no lactic acid bacteria and containing the above lactic acid bacteria (SIID21336-L1, SIID21336-L2, or SIID21336-L3) is given to tomato seedlings as fertilizer. When mature tomatoes were examined, lactic acid bacteria were detected in the tomatoes. Moreover, it was confirmed that it matured better (faster and more) than when fertilizer containing no lactic acid bacteria was given.
このように、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3）は、肥料に含有させることで、野菜等の植物の内部で増殖し、肥料を用いて育成した植物に乳酸菌を投与することができるとともに、その実にまで含有させることができる。なお、これは、乳酸菌を含有する野菜の製造方法としても利用できる。 As described above, the lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3) grow inside a plant such as vegetables by being contained in a fertilizer, and administer the lactic acid bacteria to a plant grown using the fertilizer. It can be contained up to its fruit. This can also be used as a method for producing vegetables containing lactic acid bacteria.
（３）生菌剤
海水、河川水（淡水）、汽水（海水と河川水とが混合したもの）、軽質石油を添加した海水、海底土について、乳酸菌の含有状態を調べたところ、いずれからも乳酸菌が検出されなかった。
(3) Live bacteria agent When we examined the content of lactic acid bacteria in seawater, river water (freshwater), brackish water (mixed of seawater and river water), seawater to which light petroleum was added, and seabed soil, Lactic acid bacteria were not detected.
この乳酸菌が検出されない海水、河川水、汽水、軽質石油を添加した海水、海底土に市販されている代表的な乳酸菌（ヨーグルト）を添加し放置したところ、添加した乳酸菌が死滅することを確認した。 Seawater, river water, brackish water, seawater to which light petroleum was added, and representative lactic acid bacteria (yogurt) marketed on the seabed soil were added and left to stand, and it was confirmed that the added lactic acid bacteria were killed. .
しかしながら、乳酸菌が検出されない海水、河川水、汽水、軽質石油を添加した海水、海底土に上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）を添加し放置したところ、添加した乳酸菌が死滅せずに増殖することが確認された。 However, the above lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three) are added to seawater, river water, brackish water, seawater with light petroleum added, and seabed soil where no lactic acid bacteria are detected. When added and allowed to stand, it was confirmed that the added lactic acid bacteria grew without being killed.
さらに、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）を添加した海水で養殖された生海苔を保存したところ、海苔から乳酸菌が検出された。 Furthermore, when the raw nori cultured in seawater to which the above lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three) was added was preserved, lactic acid bacteria were detected from the nori .
また、上記（２）の肥料でも確認されているように、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）を添加した土でも乳酸菌が死滅することはなかった。 In addition, as confirmed in the fertilizer of (2) above, the lactic acid bacteria were killed even in the soil added with the lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three). I never did.
そのため、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）は、水や土などとともに乳酸菌を投与することができ、しかも、植物・動物・菌類等と人間とのつながりを一体に捉えたBIOTOP（ビオトープ）の保護・保全・復元・創出である環境対策に生きた乳酸菌を投与することができる生菌剤として利用することができる。 Therefore, the lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three) can be administered with lactic acid bacteria together with water, soil, etc., and plants, animals and fungi It can be used as a viable bacterial agent that can administer live lactic acid bacteria for environmental measures that protect, preserve, restore and create BIOTOP (biotope) that captures the connection between humans and others.
（４）飲食物
上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）を疑似胃酸に添加して放置したところ、添加した乳酸菌が死滅せずに生存することが確認された。
(4) Food and drink When the above lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three) are added to pseudo gastric acid and left to stand, the added lactic acid bacteria are not killed. It was confirmed to survive.
そのため、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）は、飲食物に添加することができる。なお、同様に、上記乳酸菌（SIID21336-L1、SIID21336-L2、SIID21336-L3のいずれか１種又は２種若しくは３種）は、医薬品や健康補助食品などにも添加することができる。 Therefore, the lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three) can be added to food and drink. Similarly, the lactic acid bacteria (SIID21336-L1, SIID21336-L2, SIID21336-L3, one or two or three) can be added to pharmaceuticals, health supplements, and the like.
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| JP2009100677A (en) * | 2007-10-23 | 2009-05-14 | Daiwarandam Co Ltd | Fruit and vegetable, and method for cultivating the fruit and vegetable |
| JP2009225792A (en) * | 2008-02-29 | 2009-10-08 | Sachiko Tsugeta | New lactic acid bacterial strain, and use thereof |
| JP2016123382A (en) * | 2015-01-07 | 2016-07-11 | 株式会社かつらぎ産業 | Lactic acid bacteria, and production method of fermentation product |
| JP5958985B1 (en) * | 2015-12-04 | 2016-08-02 | 上坂 喜美枝 | Lactic acid bacteria solution, fermented product production method, edible plant production method, lactic acid bacteria solution production method |
| JP6045743B1 (en) * | 2016-09-13 | 2016-12-14 | 株式会社新栄設計事務所 | Lactic acid bacteria and feed / fertilizer / viable bacteria containing the same |
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| JP2009100677A (en) * | 2007-10-23 | 2009-05-14 | Daiwarandam Co Ltd | Fruit and vegetable, and method for cultivating the fruit and vegetable |
| JP2009225792A (en) * | 2008-02-29 | 2009-10-08 | Sachiko Tsugeta | New lactic acid bacterial strain, and use thereof |
| JP2016123382A (en) * | 2015-01-07 | 2016-07-11 | 株式会社かつらぎ産業 | Lactic acid bacteria, and production method of fermentation product |
| JP5958985B1 (en) * | 2015-12-04 | 2016-08-02 | 上坂 喜美枝 | Lactic acid bacteria solution, fermented product production method, edible plant production method, lactic acid bacteria solution production method |
| JP6045743B1 (en) * | 2016-09-13 | 2016-12-14 | 株式会社新栄設計事務所 | Lactic acid bacteria and feed / fertilizer / viable bacteria containing the same |
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