JPH06217713A - Lactic bacteria preparation for silage production - Google Patents
Lactic bacteria preparation for silage productionInfo
- Publication number
- JPH06217713A JPH06217713A JP4355857A JP35585792A JPH06217713A JP H06217713 A JPH06217713 A JP H06217713A JP 4355857 A JP4355857 A JP 4355857A JP 35585792 A JP35585792 A JP 35585792A JP H06217713 A JPH06217713 A JP H06217713A
- Authority
- JP
- Japan
- Prior art keywords
- lactobacillus
- lactic acid
- strain
- sbt
- salivarius
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 241000894006 Bacteria Species 0.000 title claims abstract description 73
- 238000002360 preparation method Methods 0.000 title claims abstract description 39
- 239000004460 silage Substances 0.000 title claims abstract description 34
- 238000004519 manufacturing process Methods 0.000 title abstract description 13
- 241000186869 Lactobacillus salivarius Species 0.000 claims abstract description 15
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 138
- 239000004310 lactic acid Substances 0.000 claims description 71
- 235000014655 lactic acid Nutrition 0.000 claims description 69
- 241000186660 Lactobacillus Species 0.000 claims description 32
- 229940039696 lactobacillus Drugs 0.000 claims description 28
- 244000199866 Lactobacillus casei Species 0.000 claims description 26
- 235000013958 Lactobacillus casei Nutrition 0.000 claims description 24
- 229940017800 lactobacillus casei Drugs 0.000 claims description 24
- 238000011160 research Methods 0.000 claims description 7
- 240000008042 Zea mays Species 0.000 abstract description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 abstract description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 abstract description 5
- 235000005822 corn Nutrition 0.000 abstract description 5
- 241000209140 Triticum Species 0.000 abstract description 4
- 235000021307 Triticum Nutrition 0.000 abstract description 4
- 238000001035 drying Methods 0.000 abstract description 4
- 235000013312 flour Nutrition 0.000 abstract description 4
- 241000218588 Lactobacillus rhamnosus Species 0.000 abstract description 3
- 229930006000 Sucrose Natural products 0.000 abstract description 3
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 abstract description 3
- 239000005720 sucrose Substances 0.000 abstract description 3
- 230000002062 proliferating effect Effects 0.000 abstract description 2
- 241001147746 Lactobacillus delbrueckii subsp. lactis Species 0.000 abstract 1
- 230000035755 proliferation Effects 0.000 abstract 1
- 238000012937 correction Methods 0.000 description 20
- 238000000034 method Methods 0.000 description 18
- 239000002609 medium Substances 0.000 description 17
- 239000000463 material Substances 0.000 description 12
- 244000025254 Cannabis sativa Species 0.000 description 11
- 238000000855 fermentation Methods 0.000 description 9
- 230000004151 fermentation Effects 0.000 description 9
- 238000003860 storage Methods 0.000 description 9
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 8
- 230000012010 growth Effects 0.000 description 8
- 238000012258 culturing Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 235000015140 cultured milk Nutrition 0.000 description 6
- 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 5
- 239000004459 forage Substances 0.000 description 5
- 239000008103 glucose Substances 0.000 description 5
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 230000036962 time dependent Effects 0.000 description 4
- 240000004296 Lolium perenne Species 0.000 description 3
- 240000004658 Medicago sativa Species 0.000 description 3
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000010902 straw Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 241000304886 Bacilli Species 0.000 description 2
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 2
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 2
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 2
- 240000006024 Lactobacillus plantarum Species 0.000 description 2
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 2
- 241000192132 Leuconostoc Species 0.000 description 2
- 241000192001 Pediococcus Species 0.000 description 2
- 241000209504 Poaceae Species 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 244000057717 Streptococcus lactis Species 0.000 description 2
- 235000014897 Streptococcus lactis Nutrition 0.000 description 2
- 230000009603 aerobic growth Effects 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 229910000019 calcium carbonate Inorganic materials 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 229940072205 lactobacillus plantarum Drugs 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 240000004585 Dactylis glomerata Species 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 241000194041 Lactococcus lactis subsp. lactis Species 0.000 description 1
- 241001317416 Lius Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 240000006394 Sorghum bicolor Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 235000014969 Streptococcus diacetilactis Nutrition 0.000 description 1
- 241000194049 Streptococcus equinus Species 0.000 description 1
- 241000219793 Trifolium Species 0.000 description 1
- 239000005862 Whey Substances 0.000 description 1
- 102000007544 Whey Proteins Human genes 0.000 description 1
- 108010046377 Whey Proteins Proteins 0.000 description 1
- 229910021536 Zeolite Inorganic materials 0.000 description 1
- 238000010564 aerobic fermentation Methods 0.000 description 1
- 235000013527 bean curd Nutrition 0.000 description 1
- 235000013405 beer Nutrition 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 230000002301 combined effect Effects 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000003977 dairy farming Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000006458 gyp medium Substances 0.000 description 1
- 229940059442 hemicellulase Drugs 0.000 description 1
- 108010002430 hemicellulase Proteins 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
- 235000021419 vinegar Nutrition 0.000 description 1
- 239000010457 zeolite Substances 0.000 description 1
Landscapes
- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、牧草、飼料作物のサイ
レージ調製用新規乳酸菌製剤に関する。更に詳しくは、
ラクトバチルス属細菌からなるサイレージ調製用乳酸菌
製剤に関する。TECHNICAL FIELD The present invention relates to a novel lactic acid bacterium preparation for silage preparation of grass and forage crops. For more details,
The present invention relates to a lactic acid bacterium preparation for silage preparation, which comprises a bacterium belonging to the genus Lactobacillus.
【０００２】[0002]
【従来の技術】サイレージとは牧草、飼料作物を嫌気的
条件下で乳酸発酵させたもので、牧草、飼料作物の貯蔵
のために利用されている。良質のサイレージを調製する
上で重要なことは、好気発酵を抑制すること及び酪酸発
酵を阻止することである。前者を達成するには、サイレ
ージ原料をサイロに詰めたとき早期に嫌気状態にすれば
よく、これはサイロの密封を素早く完全に行うことによ
ってある程度解決できる。後者の酪酸発酵の抑制は、酪
酸発酵菌の増殖を抑制することによって行うことができ
る。酪酸菌はサイレージのｐＨが４．２以下ではその活
動が抑制されることから、サイレージの乳酸発酵が速や
かに進行しその生産する乳酸によってｐＨが速やかに下
がることが重要である。サイレージの乳酸発酵に与る乳
酸菌としては、トウモロコシサイレージから単離された
例としてラクトバチルス・プランタルム（Lactobacillu
s plantarum）、ラクトバチルス・カゼイ（Lactobacil
lus casei)、ラクトバチルス・ブレビス（Lactobacillu
s brevis）などのラクトバチルス属細菌、ストレプトコ
ッカス・ラクチス（Streptococcus lactis）やストレプ
トコッカス・ボビス（Storeptococcus bovis）などのス
トレプトコッカス属細菌、ロイコノストック（Leuconos
toc）属細菌、ペディオコッカス(Pediococcus) 属細菌
などが知られている（ジャーナル・オブ・アプライド・
バクテリオロジー（Journal of Applied Bacteriology)
第６０巻、８３−９２頁、１９８６年）。これらの中で
も、乳酸菌発酵を優勢にし良質のサイレージを調製する
には、乳酸菌はホモ型発酵菌であることが好ましい。従
来、酪酸菌をはじめ、カビ、酵母など良質サイレージ調
製に好ましくない菌の増殖を阻止し、速やかに効率のよ
い乳酸発酵を促すために、サイレージ調製時に種々の乳
酸菌を添加することが行われている。例えば、特開平1-
211487号公報にはラクトバチルス・カゼイの添加が、特
公平1-18716号公報にはラクトバチルス・プランタルム
類縁菌が、特公平3-34909号公報にはラクトバチルス・
キシローサス（Lactobacillus xylosus）が、それぞれ
サイレージ調製用乳酸菌として開示されている。また、
ラクトバチルス属菌としては、特開平2-257875号、特開
平3-285674号および特開平3-285675号公報にも開示され
ている。BACKGROUND OF THE INVENTION Silage is obtained by lactic acid fermenting grass and forage crops under anaerobic conditions, and is used for storage of grass and forage crops. In order to prepare good quality silage, it is important to suppress aerobic fermentation and prevent butyric acid fermentation. To achieve the former, the silage material can be anaerobic prematurely when it is packed into the silo, which can be solved to some extent by quickly and completely sealing the silo. The latter suppression of butyric acid fermentation can be performed by suppressing the growth of butyric acid-fermenting bacteria. Since the activity of butyric acid bacteria is suppressed when the pH of silage is 4.2 or less, it is important that the lactic acid fermentation of silage rapidly progresses and the pH is rapidly lowered by the lactic acid produced. Lactobacillus plantarum ( Lactobacillus) is an example of lactic acid bacteria involved in lactic acid fermentation of silage.
s plantarum ), Lactobacil
lus casei) , Lactobacillus
s brevis) Lactobacillus bacteria such as Streptococcus lactis (Streptococcus lactis) and Streptococcus bovis (Storeptococcus bovis) Streptococcus bacteria such as, Leuconostoc (Leuconos
toc ) and Pediococcus (Pediococcus ) bacteria are known (Journal of Applied
Bacteriology (Journal of Applied Bacteriology)
60, 83-92, 1986). Among these, the lactic acid bacterium is preferably a homofermentative bacterium in order to make lactic acid bacterium fermentation dominant and to prepare good quality silage. Conventionally, various lactic acid bacteria have been added at the time of silage preparation in order to prevent the growth of butyric acid bacteria, fungi, yeast, and other bacteria that are not preferable for high-quality silage preparation, and promptly promote efficient lactic acid fermentation. There is. For example, Japanese Patent Laid-Open No. 1-
No. 211487 discloses addition of Lactobacillus casei, Japanese Patent Publication No. 1-18716 discloses Lactobacillus plantarum related bacteria, and Japanese Patent Publication No. 3-34909 discloses Lactobacillus casei.
Lactobacillus xylosus is disclosed as a lactic acid bacterium for silage preparation. Also,
Lactobacillus bacteria are also disclosed in JP-A-2-257875, JP-A-3-285674 and JP-A-3-285675.
【０００３】[0003]
【発明が解決しようとする課題】これらの乳酸菌を使用
しても、原料草の状態やサイロの密封状態などによって
は他の微生物を抑えられず必ずしも良質なサイレージ調
製がなされていない場合があり、もっと強力でより速や
かに増殖しｐＨを急速に下げるサイレージ調製用乳酸菌
が望まれていた。また、九州地方など夏場気温の高い酪
農地帯では現在の乳酸菌製剤ではその作用が十分発揮さ
れない場合もあり、高温でも活性を維持している乳酸菌
の添加が望まれている。本発明は、従来の乳酸菌よりも
非常に短時間で増殖し、しかも比較的高温環境下でも有
効に働くサイレージ用乳酸菌を提供することを目的とす
るものである。Even if these lactic acid bacteria are used, it may not be possible to suppress other microorganisms depending on the condition of the raw material grass or the sealed condition of the silo, so that good quality silage may not always be prepared. Lactic acid bacteria for silage preparation have been desired, which are more potent, grow more rapidly, and lower the pH rapidly. Further, in a dairy farming area where the temperature is high in the summer such as Kyushu region, there are cases where the action of the present lactic acid bacterium preparation is not sufficiently exerted, and therefore addition of lactic acid bacterium which maintains the activity even at high temperature is desired. It is an object of the present invention to provide a lactic acid bacterium for silage that grows in a much shorter time than conventional lactic acid bacteria, and that works effectively even in a relatively high temperature environment.
【０００４】[0004]
【課題を解決するための手段】本発明者は、上記の問題
点を克服するべく鋭意研究を行った結果、ラクトバチル
ス属桿菌でホモ型乳酸発酵を行う細菌の中でも発酵乳な
どに含まれているラクトバチルス・カゼイ・サブスペシ
ーズ・ラムノーサス（Lactobacillus casei subsp. rha
mnosus）、消化管中などに見いだされるラクトバチルス
・サリバリウス・サブスピーシーズ・サリバリウス（La
ctobacillus salivalius subsp. salivarius）および発
酵乳などに見いだされるラクトバチルス・デルブロッキ
ー・サブスピーシーズ・ラクチス（Lactobacillus delb
rueckii subsp. lactis）の添加によって意外にも上記
の目的が達成されることを見いだし、本発明を完成する
に至った。即ち、本発明によれば、ラクトバチルス属の
ホモ型乳酸発酵桿菌であるラクトバチルス・カゼイ・サ
ブスペシーズ・ラムノーサス（Lactobacillus casei su
b-sp. rhamnosus)、ラクトバチルス・サリバリウス・サ
ブスピーシーズ・サリバリウス（Lactobacillus saliva
lius subsp. salivarius）及びラクトバチルス・デルブ
ロッキー・サブスピーシーズ・ラクチス（Lactobacillu
s delbrueckii sub-sp. lactis) の少なくとも１種から
なるサイレージ調製用乳酸菌製剤が提供される。本発明
では、ラクトバチルス・カゼイ・サブスピーシーズ・ラ
ムノサス、ラクトバチルス・サリバリウスは各々単独で
使用してもよいし、併用してもよい。本発明で使用でき
るラクトバチルス・カゼイ・サブスピーシーズ・ラムノ
ーサスの菌株については特に限定されないが、好ましい
例としてSBT-2300株（微工研菌寄第１３２４５号）が挙
げられる。また、本発明で使用できるラクトバチルス・
サリバリウス・サブスピーシーズ・サリバリウスの菌株
についても特に限定されないが、好ましい例としてSBT-
2670株（微工研菌寄第１３２４７号）が挙げられる。ま
た、本発明で使用できるラクトバチルス・デルブロッキ
ー・サブスペシーズ・ラクチスの菌株についても特に限
定されないが、好ましい例としてSBT-2664株（微工研菌
寄第１３２４６号）が挙げられる。これらの菌株は、発
酵乳及び乳児糞便から単離されたものであって、いずれ
も次の形態的性状を示す。 (1) 巾０．５〜１．１μm 、長さ２〜７μm の桿菌 (2) グラム染色性；陽性 (3) 芽胞の形成；なし (4) 運 動 性；なし (5) 好気性生育能及び嫌気性生育能を有する また、これらの菌株は、次の培地でよく生育し、ＢＬ寒
天培地で３５℃で、３日間嫌気培養するとコロニーを形
成する。また、これらの菌株の生理的性質を表１に示
す。Means for Solving the Problems As a result of intensive studies to overcome the above-mentioned problems, the present inventor has found that among fermented milk and the like among bacteria that perform homo-lactic acid fermentation with Lactobacillus bacilli. Lactobacillus casei subsp. Rha
mnosus ), Lactobacillus salivarius, subspecies salivarius ( La ) found in digestive tract
ctobacillus salivalius subsp. salivarius ) and fermented milk found in Lactobacillus delbrocky subspecies lactis ( Lactobacillus delb)
It was found that the above object was unexpectedly achieved by the addition of rueckii subsp. lactis ), and the present invention was completed. That is, according to the present invention, Lactobacillus casei subspecies rhamnosus which is a homo-type lactic acid fermenting bacillus of the genus Lactobacillus
b-sp. rhamnosus) , Lactobacillus saliva ( Lactobacillus saliva)
lius subsp. salivarius ) and Lactobacillus del Blocky Subspecies Lactobacillus
s delbrueckii sub-sp. lactis ). In the present invention, Lactobacillus casei subspecies rhamnosus and Lactobacillus salivarius may be used alone or in combination. The strain of Lactobacillus casei subspecies rhamnosus that can be used in the present invention is not particularly limited, but a preferable example thereof is SBT-2300 strain (Microtechnology Research Institute No. 13245). In addition, Lactobacillus that can be used in the present invention
The strain of Salivarius subspecies Salivarius is not particularly limited, but as a preferred example, SBT-
The 2670 strain (Microtechnology Research Institute, No. 13247) can be mentioned. The strain of Lactobacillus delbrocky subspecies lactis that can be used in the present invention is also not particularly limited, but a preferable example thereof is SBT-2664 strain (Ministry of Industrial Research, No. 13246). These strains were isolated from fermented milk and infant feces, and all have the following morphological characteristics. (1) Bacilli with a width of 0.5 to 1.1 μm and a length of 2 to 7 μm (2) Gram stainability; positive (3) spore formation; none (4) mobility; none (5) aerobic growth ability Also, these strains grow well in the following medium and form colonies when anaerobically cultivated on BL agar medium at 35 ° C. for 3 days. The physiological properties of these strains are shown in Table 1.
【０００５】[0005]
【表１】 [Table 1]
【０００６】本発明の乳酸菌製剤は次のようにして製造
することができる。上記の乳酸菌の培養増殖は、通常の
乳酸菌用の培地を用いて行うことができる。培地として
は、通常乳酸菌の培養に用いられる培地であれば特に限
定されないが、例えばＧＹＰ培地（小崎道雄ら(1992)、
乳酸菌実験マニュアル、朝倉書店刊）、ＭＲＳ培地（de
Man, J.C. et al.(1960), Journal of Applied Bacte
riology, Vol.23, 130-135）や特公平4-49382号公報に
記載のルーサン煮汁培地を使用することができる。培養
条件については特に限定されないが、通常２５〜４５℃
で培養することができる。培養時間は増殖スケールによ
って左右されるが、本発明の乳酸菌は非常に増殖速度が
速く、通常１２〜２４時間で定常期に達するので、他の
乳酸菌製剤よりも比較的短時間で製造することができ
る。培養増殖した乳酸菌は、基材とともに凍結乾燥ある
いは基材に吸着乾燥して製剤化することができる。凍結
乾燥用基材としては、例えば、デキストリン、デンプ
ン、脱脂粉乳や炭酸カルシウムなどが挙げられ、吸着乾
燥用基材としてはブドウ糖などの糖類や炭酸カルシウ
ム、ゼオライト、小麦粉、ふすま、脱脂粉乳、ホエー、
デンプン、コーングリッツなどが挙げられるがこれらに
限定されるものではない。基材の量については特に限定
的ではないが、製剤中の菌数が極端に少ないとサイレー
ジ原料草に添加すべき製剤の量が多くなりすぎるし乳酸
菌の安定性の面からも好ましくない。通常、製剤１ｇあ
たり１０7 以上の乳酸菌を含有するようにするのが好ま
しい。菌数の上限は限定されないが、製造上吸着乾燥で
は通常１０9 個／ｇ、凍結乾燥では１０11個／ｇくらい
である。The lactic acid bacteria preparation of the present invention can be manufactured as follows. The culture and growth of the above-mentioned lactic acid bacterium can be carried out using an ordinary medium for lactic acid bacterium. The medium is not particularly limited as long as it is a medium usually used for culturing lactic acid bacteria, and examples thereof include GYP medium (Michio Kozaki et al. (1992),
Lactic acid bacteria experiment manual, published by Asakura Shoten, MRS medium (de
Man, JC et al. (1960), Journal of Applied Bacte
Riology, Vol.23, 130-135) and Japanese Patent Publication No. 4-49382 can be used. Culture conditions are not particularly limited, but usually 25 to 45 ° C.
Can be cultured in. Although the culturing time depends on the growth scale, the lactic acid bacterium of the present invention has a very high growth rate and usually reaches a stationary phase in 12 to 24 hours, and therefore it can be produced in a relatively shorter time than other lactic acid bacterium preparations. it can. The lactic acid bacterium that has grown in culture can be formulated together with the base material by freeze-drying or adsorption-drying on the base material. Examples of the freeze-drying base include, for example, dextrin, starch, skim milk powder and calcium carbonate, and examples of the adsorbing-drying base include sugars such as glucose and calcium carbonate, zeolite, wheat flour, bran, skim milk powder, whey,
Examples thereof include, but are not limited to, starch and corn grits. The amount of the base material is not particularly limited, but if the number of bacteria in the formulation is extremely small, the amount of the formulation to be added to the silage raw material grass becomes too large, and it is not preferable from the viewpoint of stability of lactic acid bacteria. Usually, it is preferable to contain 10 7 or more lactic acid bacteria per 1 g of the preparation. Although the upper limit of the number of bacteria is not limited, it is usually 10 9 cells / g for adsorption drying and about 10 11 cells / g for freeze-drying.
【０００７】本発明の乳酸菌製剤は、種々の牧草、飼料
作物のサイレージ調製に使用することができる。適用で
きる牧草、飼料作物の種類は特に限定されないが、オー
チャードグラス、チモシー、クローバ、アルファルフ
ァ、イタリアンライグラスなどの牧草、トウモロコシ、
ソルガムなどの飼料作物、稲わら、麦わらなどの藁類、
ビール粕、豆腐粕、酢粕などの食品製造残渣などが挙げ
られる。本発明の乳酸菌製剤は、これらのサイレージ材
料１ｇあたり菌数１０4 〜１０7 個になるように添加す
る。上述のように製剤１ｇあたり１０7 〜１０11個の乳
酸菌を含有するので、材料に添加する場合は上記の添加
量になるように加減して材料に散布すればよい。材料に
添加する場合は、粉状で、あるいは適当な容量の水に溶
解して散布することができる。必要に応じて、セルラー
ゼ、ヘミセルラーゼなどの酵素と併用することもでき
る。The lactic acid bacteria preparation of the present invention can be used for silage preparation of various grasses and feed crops. The types of grass and forage crops that can be applied are not particularly limited, but grasses such as orchard grass, timothy, clover, alfalfa and Italian ryegrass, corn,
Forage crops such as sorghum, straws such as rice straw and straw,
Examples include food manufacturing residues such as beer lees, tofu lees, and vinegar lees. The lactic acid bacteria preparation of the present invention is added so that the number of bacteria is 10 4 to 10 7 per 1 g of these silage materials. As described above, since 10 7 to 10 11 lactic acid bacteria are contained per 1 g of the preparation, when added to the material, the amount may be adjusted to the above-mentioned addition amount and sprinkled on the material. When it is added to the material, it can be sprinkled in powder form or dissolved in an appropriate volume of water. If necessary, it can be used in combination with an enzyme such as cellulase or hemicellulase.
【０００８】[0008]
製造例１ ラクトバチルス・カゼイ・サブスピーシーズ・ラムノー
サスSBT-2300株及びラクトバチルス・サリバリウス・サ
ブスピーシーズ・サリバリウスSBT-2670株それぞれブリ
ックス リバー ブロス培地（以下、ＢＬＢ培地と略
す）に接種し、３５℃で１６時間培養を行った。得られ
た培養物のそれぞれ０．２ｍｌを２０ｍｌのルーサン煮
汁培地（特開昭６４−１０９８１号公報に記載の方法に
より調製）に接種し、２４時間、３５℃で培養後、更に
その全量を２ｌのルーサン煮汁培地に接種し同じ条件で
培養した。これに同量の２４％ショ糖溶液を混合後、流
動乾燥機（大河原製作所製、ＦＢＳ−５型）でコーング
リッツ又は小麦粉に吸着させて乾燥し、各々の乳酸菌製
剤を得た。Production Example 1 Lactobacillus casei subspecies rhamnosus SBT-2300 strain and Lactobacillus salivarius subspecies salivarius SBT-2670 strain were inoculated into Brix river broth medium (hereinafter abbreviated as BLB medium) at 35 ° C. Culture was performed for 16 hours. 0.2 ml of each of the obtained cultures was inoculated into 20 ml of Rusan's broth medium (prepared by the method described in JP-A-64-10981), and after culturing at 35 ° C. for 24 hours, the total amount was 2 liters. Was inoculated into the Roussin broth medium and cultured under the same conditions. After mixing the same amount of 24% sucrose solution with this, it was adsorbed on Corn grits or wheat flour with a fluid dryer (Ogawara Seisakusho, FBS-5 type) and dried to obtain each lactic acid bacteria preparation.
【０００９】製造例２ 製造例１で得られる培養物を更に３０ｌのルーサン煮汁
培地に接種して培養後、限外ろ過膜（アミコン社製、限
外分子量１０万）で３ｌまで濃縮し、同量の２４％ショ
糖溶液を混合後、製造例１と同様の方法でコーングリッ
ツ又は小麦粉に吸着乾燥させた。 製造例３ 製造例１で得られた培養物を特開昭６４−１０９８１号
公報記載の方法で乾燥させた。すなわち、製造例１で得
られた培養物を流動乾燥機を用い、吸入温度５０℃、排
気温度２５〜２８℃で５分間乾燥し、水分６．４％の乾
燥粉末を得た。Production Example 2 The culture obtained in Production Example 1 was further inoculated into 30 liters of Rusan's broth medium, cultivated, and then concentrated to 3 liters with an ultrafiltration membrane (Amicon Co., ultramolecular weight 100,000). After mixing the amount of 24% sucrose solution, it was adsorbed and dried on the corn grits or the wheat flour by the same method as in Production Example 1. Production Example 3 The culture obtained in Production Example 1 was dried by the method described in JP-A No. 64-10981. That is, the culture obtained in Production Example 1 was dried for 5 minutes at a suction temperature of 50 ° C and an exhaust temperature of 25 to 28 ° C using a fluid dryer to obtain a dry powder having a water content of 6.4%.
【００１０】[0010]
【実施例１】培養試験 ラクトバチルス・カゼイ・サブスピーシーズ・ラムノー
サスSBT-2300株、ラクトバチルス・サリバリウス・サブ
スピーシーズ・サリバリウスSBT-2670株、ラクトバチル
ス・デルブロッキー・サブスピーシーズ・ラクチスSBT-
2664株それぞれブリックス リバー ブロス培地（以
下、ＢＬＢ培地と略す）に接種し、３５℃で１６時間培
養を行った。培養物を特開昭６４−１０９８１号公報に
記載の２％ブドウ糖添加ルーサン培地に１％（ｖ／ｖ）
接種し、３８℃で１６時間前培養した。得られた培養物
を更に２％ブドウ糖添加ルーサン培地に１％（ｖ／ｖ）
接種し、３８℃で培養した。接種直後（０時間後）、
４、８、１２、１６、２０及び２４時間後に培養試料を
採取しｐＨ、酸度、生菌数を測定した。ｐＨは１０℃下
でガラス電極ｐＨメーターにより測定した。酸度（％）
は各試料９．０ｇを蒸留水で３倍に希釈し、０．１Ｎ水
酸化ナトリウム溶液でｐＨ８．０（ｐＨメーターによる
確認）に滴定した量から乳酸等量（％）を求めた。生菌
数は生理食塩水を希釈液に用い、平板希釈法により測定
した。対照として市販のサイレージ添加用乳酸菌スノー
ラクトＬ（雪印種苗株式会社製）の成分であるラクトバ
チルス・カゼイSBT-2232株を使用して同様に行った。結
果を表２及び図１〜３に示す。これらの結果から明らか
なように本発明の乳酸菌はいずれも従来のサイレージ用
乳酸菌であるラクトバチルス・カゼイよりも初期の増殖
速度が早く、ｐＨの低下及び乳酸の生成が速いことが判
明した。[Example 1] Culture test Lactobacillus casei subspecies rhamnosus SBT-2300 strain, Lactobacillus salivarius subspecies salivarius SBT-2670 strain, Lactobacillus delbrocky subspecies lactis SBT-
Each of the 2664 strains was inoculated into a Brix River Broth medium (hereinafter abbreviated as BLB medium), and cultured at 35 ° C. for 16 hours. 1% (v / v) of the culture was added to 2% glucose-added Luthan's medium described in JP-A-64-10981.
The cells were inoculated and precultured at 38 ° C for 16 hours. The obtained culture was further added to 2% glucose-containing Luthan's medium at 1% (v / v).
Inoculated and cultured at 38 ° C. Immediately after inoculation (0 hours later),
Culture samples were collected after 4, 8, 12, 16, 20, and 24 hours, and pH, acidity, and viable cell count were measured. The pH was measured with a glass electrode pH meter at 10 ° C. acidity(%)
For each sample, 9.0 g of each sample was diluted three times with distilled water, and the lactic acid equivalent amount (%) was determined from the amount titrated to pH 8.0 (confirmation with a pH meter) with a 0.1N sodium hydroxide solution. The viable cell count was measured by the plate dilution method using physiological saline as a diluent. As a control, Lactobacillus casei SBT-2232 strain, which is a component of commercially available lactic acid bacterium Snowlact L for silage addition (manufactured by Snow Brand Seed Co., Ltd.) was used. The results are shown in Table 2 and FIGS. As is clear from these results, it was found that all of the lactic acid bacteria of the present invention had an initial growth rate faster than that of the conventional lactic acid bacterium for silage, Lactobacillus casei, lowering of pH and production of lactic acid.
【００１１】[0011]
【表２】 [Table 2]
【００１２】[0012]
【実施例２】培養試験 実施例１と同様の方法で前培養したラクトバチルス・カ
ゼイ・サブスペシーズ・ラムノーサスSBT-2300株、ラク
トバチルス・サリバリウス・サブスペシーズ・サリバリ
ウスSBT-2670株、ラクトバチルス・デブロッキー・サブ
スペシーズ・ラクチスSBT-2664株それぞれを、培養温度
を３５℃及び４５℃の２つの条件に設定する以外は実施
例１と同様の方法で培養し、８、１６及び３２時間後に
培養物試料を採取しｐＨと酸度（％）を測定した。対照
として実施例１と同じラクトバチルス・カゼイSBT-2232
株を使用した。結果を表３に示す。結果から明らかなよ
うに本発明の乳酸菌は４５℃の高温環境下でも菌の活性
は高く、従来のラクトバチルス・カゼイよりもｐＨが速
やかに低下し乳酸量（酸度）が増加した。Example 2 Culture test Lactobacillus casei subspecies rhamnosus SBT-2300 strain, Lactobacillus salivarius subspecies salivarius SBT-2670 strain, and Lactobacillus debu precultured in the same manner as in Example 1 Each Rocky subspecies lactis SBT-2664 strain was cultured in the same manner as in Example 1 except that the culture temperature was set to two conditions of 35 ° C. and 45 ° C., and after 8 and 16 and 32 hours, the culture A sample was taken and pH and acidity (%) were measured. As a control, the same Lactobacillus casei SBT-2232 as in Example 1
Strains were used. The results are shown in Table 3. As is clear from the results, the lactic acid bacterium of the present invention has a high microbial activity even in a high temperature environment of 45 ° C., the pH was rapidly lowered and the amount of lactic acid (acidity) was increased as compared with the conventional Lactobacillus casei.
【００１３】[0013]
【表３】 [Table 3]
【００１４】[0014]
【実施例３】サイレージ添加試験（アルファルファ ） 収穫したアルファルファ材料草を軽く予乾し、マウント
カッターで１５〜２５ｍｍに切断後、製造例２で製造し
た乳酸菌製剤をそれぞれ下記の量添加しブドウ糖を０．
５５％加えて良く混合し、１ｌの実験ポリサイロに詰め
込んだ。同じサイレージ調製を複数行い、各乳酸菌製剤
について１５、２５、３５、４５℃の温度条件で３２日
間貯蔵した。貯蔵期間中、ラクトバチルス・カゼイ・サ
ブスピーシーズ・ラムノーサスSBT-2300株の製剤を貯蔵
温度３５℃の条件で使用したものについては貯蔵１、
２、４、８、１６日後に試料を採取しｐＨを測定した。
また、全ての試料について３２日後にｐＨを測定した。
対照として市販のサイレージ用乳酸菌製剤スノーラクト
Ｌ（雪印種苗株式会社製）を添加したもの（SBT-2232
区）、乳酸菌を吸着させていない基材とブドウ糖のみを
添加したもの（無接種区）および何も添加しないもの
（無添加区）を調製し同様の試験を行った。 乳酸菌製剤添加量 SBT-2300株 ６．９×１０5 ｃｆ
ｕ／材料草ｇ SBT-2232株 ２×１０5 ｃｆｕ／材料草ｇ 結果を表４及び５に示す。表４の結果から明らかなよう
に本発明の各乳酸菌製剤は１５℃の低温から４５℃の高
温まで広い温度域で有効に働く。また表４から明らかな
ように本発明の乳酸菌製剤は従来の乳酸菌製剤に比較し
てサイレージ調製後速やかにｐＨを下げるものである。[Example 3] Silage addition test (alfalfa ) The harvested alfalfa material grass was lightly pre-dried and cut into 15 to 25 mm with a mount cutter, and the lactic acid bacteria preparation produced in Production Example 2 was added to each of the following amounts to give 0 glucose. ．
55% was added, mixed well and packed in 1 liter experimental polysilo. The same silage preparation was performed multiple times, and each lactic acid bacterium preparation was stored for 32 days under the temperature conditions of 15, 25, 35, and 45 ° C. During the storage period, Lactobacillus casei subspecies Rhamnosus SBT-2300 was used for storage at a storage temperature of 35 ° C.
Samples were taken after 2, 4, 8 and 16 days to measure pH.
In addition, pH was measured after 32 days for all samples.
As a control, a commercially available lactobacillus formulation for silage Snowlacto L (manufactured by Snow Brand Seed Co., Ltd.) was added (SBT-2232
Group), a substrate to which lactic acid bacteria have not been adsorbed and a substance to which only glucose was added (non-inoculation group) and one to which nothing was added (non-inoculation group) were prepared and the same test was conducted. Amount of lactic acid bacteria preparation added SBT-2300 strain 6.9 × 10 5 cf
u / grass g SBT-2232 strain 2 × 10 5 cfu / grass g The results are shown in Tables 4 and 5. As is clear from the results in Table 4, each lactic acid bacterium preparation of the present invention works effectively in a wide temperature range from a low temperature of 15 ° C to a high temperature of 45 ° C. Further, as is apparent from Table 4, the lactic acid bacterium preparation of the present invention lowers the pH immediately after the silage preparation as compared with the conventional lactic acid bacterium preparation.
【００１５】[0015]
【表４】 [Table 4]
【００１６】[0016]
【表５】 [Table 5]
【００１７】[0017]
【実施例４】サイレージ添加試験（イタリアンライグラス ） 材料草としてイタリアンライグラスを用い、乳酸菌製剤
として製造例１で調製したものを使用し、ブドウ糖は添
加しない以外は実施例３と同様の方法で材料草を実験ポ
リサイロに詰めた。それぞれの生菌数は１０5 ｃｆｕ／
材料草ｇに調製した。各ポリサイロを３０℃で貯蔵し貯
蔵直後（０日後）、１、３、３０日後に一部試料を採取
してｐＨを測定した。結果を表６に示す。表６から明ら
かなように本発明の乳酸菌製剤を用いると貯蔵後、速や
かにサイレージのｐＨを下げることができた。[Example 4] Silage addition test (Italian ryegrass ) The material grass was prepared in the same manner as in Example 3 except that Italian ryegrass was used as the material grass, the lactic acid bacteria preparation prepared in Production Example 1 was used, and glucose was not added. Was packed in an experimental poly silo. The viable cell count of each is 10 5 cfu /
The material was adjusted to g. Each polysilo was stored at 30 ° C., and immediately after storage (0 days later), 1, 3 and 30 days later, some samples were taken to measure the pH. The results are shown in Table 6. As is clear from Table 6, when the lactic acid bacterium preparation of the present invention was used, the pH of silage could be lowered immediately after storage.
【００１８】[0018]
【表６】 [Table 6]
【００１９】[0019]
【実施例５】併用効果（培地試験 ） 実施例２と同様の方法でラクトバチルス・カゼイ・サブ
スピーシーズ・ラムノーサスSBT-2300株、ラクトバチル
ス・サリバリウス・サブスピーシーズ・サリバリウスSB
T-2670株及びラクトバチルス・デブロッキー・サブスピ
ーシーズ・ラクチスSBT-2664株を２種あるいは３種併用
して培養を行い、貯蔵温度３５℃及び４５℃において貯
蔵し、貯蔵中のｐＨと酸度（乳酸等量％）を経時的に測
定した。結果を表７及び図４〜７に示す。結果から明ら
かなように本発明の乳酸菌ラクトバチルス・カゼイ・サ
ブスピーシーズ・ラムノーサス、ラクトバチルス・サリ
バリウス・サブスピーシーズ・サリバリウスおよびラク
トバチルス・デルブロッキー・サブスピーシーズ・ラク
チスを併用することにより、３５℃及び４５℃のいずれ
の貯蔵温度でもｐＨの低下と酸度の上昇に関し単独使用
と同等あるいはそれ以上の効果を示した。[Example 5] Combined effect (medium test ) In the same manner as in Example 2, Lactobacillus casei subspecies rhamnosus SBT-2300 strain, Lactobacillus salivarius subspecies salivarius SB
T-2670 strain and Lactobacillus debrocky subspecies lactis SBT-2664 strain were cultivated by using two or three kinds in combination and stored at storage temperatures of 35 ° C. and 45 ° C., and pH and acidity during storage ( Lactic acid equivalent%) was measured over time. The results are shown in Table 7 and FIGS. As is clear from the results, the combined use of the lactic acid bacteria Lactobacillus casei subspecies rhamnosus, Lactobacillus salivarius subspecies salivarius and Lactobacillus delbrocky subspecies lactis of the present invention resulted in 35 ° C and 45 ° C. At any storage temperature of ° C, the same effect as or higher than the single use was shown with respect to the decrease of pH and the increase of acidity.
【００２０】[0020]
【表７】 [Table 7]
【００２１】[0021]
【発明の効果】本発明のサイレージ調製用乳酸菌製剤
は、従来の乳酸菌に比較して非常に増殖が速く従来の乳
酸菌よりも速やかにサイレージのｐＨを下げて他の好ま
しくない菌の増殖を抑えることができ、それによって良
質のサイレージを調製することができる。また、この増
殖活性は環境温度が４５℃の高温でもほとんど変わら
ず、寒冷地から高温地まで広い範囲でサイレージ調製に
使用することができる。INDUSTRIAL APPLICABILITY The lactic acid bacterium preparation for silage preparation of the present invention is very fast in growth as compared with conventional lactic acid bacteria, and lowers the pH of silage more quickly than conventional lactic acid bacteria to suppress the growth of other undesirable bacteria. The result is good quality silage. Further, this proliferative activity remains almost unchanged even when the environmental temperature is as high as 45 ° C., and can be used for silage preparation in a wide range from cold regions to high temperature regions.
【図面の簡単な説明】[Brief description of drawings]
【図１】本発明の乳酸菌株SBT-2300株の３８℃での培養
によるｐＨの経時的変化を示す。FIG. 1 shows a change with time in pH of a lactic acid bacterium strain SBT-2300 of the present invention due to culture at 38 ° C.
【図２】本発明の乳酸菌株SBT-2670株の３８℃での培養
によるｐＨの経時的変化を示す。FIG. 2 shows the time-dependent changes in pH of the lactic acid bacterium strain SBT-2670 of the present invention due to culture at 38 ° C.
【図３】本発明の乳酸菌株SBT-2664株の３８℃での培養
によるｐＨの経時的変化を示す。FIG. 3 shows the time-dependent change in pH of the lactic acid bacterium strain SBT-2664 of the present invention due to culture at 38 ° C.
【図４】本発明の乳酸菌株SBT-2300株とSBT-2670株とを
併用し４５℃での培養によるｐＨの経時的変化を示す。FIG. 4 shows time-dependent changes in pH due to culturing at 45 ° C. using the lactic acid bacterium strain SBT-2300 strain and SBT-2670 strain of the present invention in combination.
【図５】本発明の乳酸菌株SBT-2670株とSBT-2664株とを
併用し４５℃での培養によるｐＨの経時的変化を示す。FIG. 5 shows a change with time in pH due to culturing at 45 ° C. using the lactic acid bacterium strain SBT-2670 strain and SBT-2664 strain of the present invention in combination.
【図６】本発明の乳酸菌株SBT-2300株とSBT-2664株とを
併用し４５℃での培養によるｐＨの経時的変化を示す。[Fig. 6] Fig. 6 shows changes in pH over time due to culturing at 45 ° C using the lactic acid bacterium strain SBT-2300 strain of the present invention and SBT-2664 strain in combination.
【図７】本発明の乳酸菌株SBT-2300株とSBT-2670株とSB
T-2664株とを併用し４５℃での培養によるｐＨの経時的
変化を示す。FIG. 7: Lactobacillus strains SBT-2300 strain, SBT-2670 strain and SB of the present invention
The time-dependent change of pH by the culture | cultivation at 45 degreeC using T-2664 strain together is shown.
─────────────────────────────────────────────────────
─────────────────────────────────────────────────── ───
【手続補正書】[Procedure amendment]
【提出日】平成５年４月５日[Submission date] April 5, 1993
【手続補正１】[Procedure Amendment 1]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】請求項１[Name of item to be corrected] Claim 1
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【手続補正２】[Procedure Amendment 2]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】請求項２[Name of item to be corrected] Claim 2
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【手続補正３】[Procedure 3]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】０００４[Correction target item name] 0004
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【０００４】[0004]
【課題を解決するための手段】本発明者は、上記の問題
点を克服するべく鋭意研究を行った結果、ラクトバチル
ス属桿菌でホモ型乳酸発酵を行う細菌の中でも発酵乳な
どに含まれているラクトバチルス・カゼイ・サブスピー
シーズ・ラムノーサス（Ｌａｃｔｏｂａｃｉｌｌｕｓ
ｃａｓｅｉｓｕｂｓｐ．ｒｈａｍｎｏｓｕｓ）、消化管
中などに見いだされるラクトバチルス・サリバリウス・
サブスピーシーズ・サリバリウス（Ｌａｃｔｏｂａｃｉ
ｌｌｕｓ ｓａｌｉｖａｒｉｕｓｓｕｂｓｐ．ｓａｌｉ
ｖａｒｉｕｓ）および発酵乳などに見いだされるラクト
バチルス・デルブロッキー・サブスピーシーズ・ラクチ
ス（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｄｅｌｂｒｕｅｃｋ
ｉｉｓｕｂｓｐ．ｌｈｃｔｉｓ）の添加によって意外に
も上記の目的が達成されることを見いだし、本発明を完
成するに至った。即ち、本発明によれば、ラクトバチル
ス属のホモ型乳酸発酵桿菌であるラクトバチルス・カゼ
イ・サブスピーシーズ・ラムノーサス（Ｌａｃｔｏｂａ
ｃｉｌｌｕｓ ｃａｓｅｉｓｕｂｓｐ．ｒｈａｍｎｃｏ
ｓｕｓ）、ラクトバチルス・サリバリウス・サブスピー
シーズ・サリバリウス（Ｌａｃｔｏｂａｃｉｌｌｕｓ
ｓａｌｉｖａｌｉｕｓｓｕｂｓｐ．ｓａｌｉｖａｒｉｕ
ｓ）及びラクトバチルス・デルブロッキー・サブスピー
シーズ・ラクチス（Ｌａｃｔｏｂａｃｉｌｌｕｓｄｅｌ
ｂｒｕｅｃｋｉｉｓｕｂｓｐ．ｌａｃｔｉｓ）の少なく
とも１種からなるサイレージ調製用乳酸菌製剤が提供さ
れる。本発明では、ラクトバチルス・カゼイ・サブスピ
ーシーズ・ラムノ−サス、ラクトバチルス・サリバリウ
スは各々単独で使用してもよいし、併用してもよい。本
発明で使用できるラクトバチルス・カゼイ・サブスピー
シーズ・ラムノーサスの菌株については特に限定されな
いが、好ましい例としてＳＢＴ−２３００株（微工研菌
寄第１３２４５号）が挙げられる。また、本発明で使用
できるラクトバチルス・サリバリウス・サブスピーシー
ズ・サリバリウスの菌株についても特に限定されない
が、好ましい例としてＳＢＴ．２６７０株（微工研菌寄
第１３２４７号）が挙げられる。また、本発明で使用で
きるラクトバチルス・デルブロッキー・サブスピーシー
ズ・ラクチスの菌株についても特に限定されないが、好
ましい例としてＳＢＴ−２６６４株（微工研菌寄第１３
２４６号）が挙げられる。これらの菌株は、発酵乳及び
乳児糞便から単離されたものであって、いずれも次の形
態的性状を示す。 （１）巾０．５〜１．１μｍ、長さ２〜７μｍの桿菌 （２）グラム染色性；陽性 （３）芽胞の形成；なし （４）運 動 性；なし （５）好気性生育能及び嫌気性生育能を有する また、これらの菌株は、次の培地でよく生育し、ＢＬ寒
天培地で３５℃で、３日間嫌気培養するとコロニーを形
成する。また、これらの菌株の生理的性質を表１に示
す。Means for Solving the Problems As a result of intensive studies to overcome the above-mentioned problems, the present inventor has found that among fermented milk and the like among bacteria that perform homo-lactic acid fermentation with Lactobacillus bacilli. It is Lactobacillus casei, substring copy <br/> seeds rhamnosus (Lactobacillus
casei subsp. Rhamnosus ), Lactobacillus salivarius found in the digestive tract, etc.
Subsp. Salivarius (Lactobaci
llus salivarus subsp. sali
varius) and Lactobacillus del are found, such as in fermented milk blocky-substring pea seeds lactis (Lactobacillus delbrueck
ii subsp. Surprisingly, it was found that the above object was achieved by the addition of lactic acid ), and the present invention was completed. That is, according to the present invention, a homozygous lactic acid fermentation bacilli Lactobacillus Lactobacillus casei substring P. Seeds rhamnosus (Lactoba
cillus casei subsp. rhamnco
sus ), Lactobacillus salivarius subspecies salivarius ( Lactobacillus)
salivalius subsp. salivariu
s ) and Lactobacillus del Blocky Subspecies Lactis ( Lactobacillus del
brückii subsp. silage lactic acid bacteria preparation comprising at least one lactis) is provided. In the present invention, Lactobacillus casei subsp. Rhamnose - suspension, Lactobacillus salivarius is may be respectively used alone or in combination. The strain of Lactobacillus casei subspecies rhamnosus that can be used in the present invention is not particularly limited, but a preferred example thereof is SBT-2300 strain (Microtechnology Research Institute No. 13245). Although there is no particular limitation on Lactobacillus salivarius, substring Phi Sea <br/>strain's salivarius that can be used in the present invention, SBT preferred examples. The 2670 strain (Microtechnology Research Institute No. 13247) can be mentioned. Further, the strain of Lactobacillus delbrocky subspecies lactis that can be used in the present invention is not particularly limited, but as a preferred example, SBT-2664 strain (Microtechnology Research Institute
No. 246). These strains were isolated from fermented milk and infant feces, and all have the following morphological characteristics. (1) Bacilli having a width of 0.5 to 1.1 μm and a length of 2 to 7 μm (2) Gram stainability; positive (3) spore formation; none (4) mobility; none (5) aerobic growth ability Also, these strains grow well in the following medium and form colonies when anaerobically cultivated on BL agar medium at 35 ° C. for 3 days. The physiological properties of these strains are shown in Table 1.
【手続補正４】[Procedure amendment 4]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】００１１[Correction target item name] 0011
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【００１１】[0011]
【表２】 [Table 2]
【手続補正５】[Procedure Amendment 5]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】００１２[Correction target item name] 0012
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【００１２】[0012]
【実施例２】培養試験 実施例１と同様の方法で前培養したラクトパチルス・カ
ゼイ・サブスピーシーズ・ラムノーサスＳＢＴ−２３０
０株、ラクトバチルス・サリバリウス・サブスピーシー
ズ・サリバリウスＳＢＴ−２６７０株、ラクトバチルス
・デルブロッキー・サブスピーシーズ・ラクチスＳＢＴ
−２６６４株それぞれを、培養温度を３５℃及び４５℃
の２つの条件に設定する以外は実施例１と同様の方法で
培養し、８、１６及び３２時間後に培養物試料を採取し
ｐＨと酸度（％）を測定した。対照として実施例１と同
じラクトバチルス・カゼイＳＢＴ−２２３２株を使用し
た。結果を表３に示す。結果から明らかなように本発明
の乳酸菌は４５℃の高温環境下でも菌の活性は高く、従
来のラクトバチルス・カゼイよりもｐＨが速やかに低下
し乳酸量（酸度）が増加した。EXAMPLE 2 Rakutopachirusu casei-substring P. Seas rhamnosus SBT-230 were pre-cultured in the same manner as culturing Test Example 1
0 shares, Lactobacillus salivarius, substring copy Sea <br/>'s salivarius SBT-2670 strain, Lactobacillus Dell blocky-substring pea seeds lactis SBT
-2664 strains were cultured at 35 ° C and 45 ° C, respectively.
The culture was performed in the same manner as in Example 1 except that the above two conditions were set, and after 8, 16 and 32 hours, a sample of the culture was sampled and the pH and acidity (%) were measured. As a control, the same Lactobacillus casei SBT-2232 strain as in Example 1 was used. The results are shown in Table 3. As is clear from the results, the lactic acid bacterium of the present invention has a high microbial activity even in a high temperature environment of 45 ° C., the pH was rapidly lowered and the amount of lactic acid (acidity) was increased as compared with the conventional Lactobacillus casei.
【手続補正６】[Procedure correction 6]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】００１３[Correction target item name] 0013
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【００１３】[0013]
【表３】 [Table 3]
【手続補正７】[Procedure Amendment 7]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】００２０[Correction target item name] 0020
【補正方法】変更[Correction method] Change
【補正内容】[Correction content]
【００２０】[0020]
【表７】 [Table 7]
───────────────────────────────────────────────────── フロントページの続き (72)発明者 田中 秀俊 北海道札幌市白石区川北３条１丁目2310番 109号 (72)発明者 山下 征夫 北海道江別市野幌東町26番14号 (72)発明者 望月 英輔 埼玉県大宮市大字東新井710−50 東新井 団地３−201 (72)発明者 豊田 修次 埼玉県所沢市緑町３丁目12番５号 煉瓦館 11 202号室 ─────────────────────────────────────────────────── --- Continuation of the front page (72) Inventor Hidetoshi Tanaka, 23-1, 109, Kawakita 3-chome, Shiroishi-ku, Sapporo-shi, Hokkaido (72) Inventor Masao Yamashita 26-14, Nopporo-higashi, Ebetsu-shi, Hokkaido (72) Inventor Mochizuki Eisuke 710-50, Higashiarai, Omiya City, Saitama Prefecture 3-201 Higashiarai housing complex 201 (72) Inventor Shuji Toyoda, 3-12-5 Midoricho, Tokorozawa-shi, Saitama Brick House 11 202 Room
Claims (4)
ーズ・ラムノーサス（Lactobacillus casei subsp. rha
mnosus）、ラクトバチルス・サリバリウス・サブスピー
シーズ・サリバリウス（Lactobacillus salivalius sub
sp. salivarius）及びラクトバチルス・デルブロッキー
・サブスペシーズ・ラクチス（Lactobacillus delbruec
kii subsp. lactis)の少なくとも１種からなるサイレー
ジ調製用乳酸菌製剤。1. Lactobacillus casei subsp. Rha
mnosus ), Lactobacillus salivalius sub
sp. salivarius ) and Lactobacillus delbruec
lactic acid bacterium preparation for silage preparation comprising at least one kind of kii subsp. lactis) .
ーズ・ラムノサスがSBT-2300株（微工研菌寄第１３２４
５号）である請求項１記載の乳酸菌製剤。2. The Lactobacillus casei subspecies rhamnosus strain SBT-2300 (Ministry of Industrial Science and Technology 1324)
No. 5), The lactic acid bacterium preparation according to claim 1.
スピーシーズ・ラクチスがSBT-2664株（微工研菌寄第１
３２４６号）である請求項１記載の乳酸菌製剤。3. The Lactobacillus delbrocky subspecies lactis strain SBT-2664 (Microtechnology Research Institute
No. 3246), The lactic acid bacterium preparation according to claim 1.
ピーシーズ・サリバリウスがSBT-2670株（微工研菌寄第
１３２４７号）である請求項１記載の乳酸菌製剤。4. The lactic acid bacterium preparation according to claim 1, wherein Lactobacillus salivarius subspecies salivarius is SBT-2670 strain (Ministry of Industrial Science and Technology No. 13247).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4355857A JPH0757186B2 (en) | 1992-12-18 | 1992-12-18 | Lactic acid bacteria preparation for silage preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4355857A JPH0757186B2 (en) | 1992-12-18 | 1992-12-18 | Lactic acid bacteria preparation for silage preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH06217713A true JPH06217713A (en) | 1994-08-09 |
| JPH0757186B2 JPH0757186B2 (en) | 1995-06-21 |
Family
ID=18446087
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4355857A Expired - Lifetime JPH0757186B2 (en) | 1992-12-18 | 1992-12-18 | Lactic acid bacteria preparation for silage preparation |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0757186B2 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003529383A (en) * | 2000-04-07 | 2003-10-07 | ソシエテ デ プロデユイ ネツスル ソシエテ アノニム | Fermented protein hydrolyzate |
| JP2006042647A (en) * | 2004-08-03 | 2006-02-16 | National Agriculture & Bio-Oriented Research Organization | New lactic acid bacterial strain and method for inhibiting silage aerobic deterioration |
| JP2008035728A (en) * | 2006-08-02 | 2008-02-21 | National Agriculture & Food Research Organization | Silage preparation additive, method for preparing silage using the silage preparation additive, and silage preparation kit |
| CN112553103A (en) * | 2020-10-10 | 2021-03-26 | 重庆市畜牧科学院 | Bullwhip grass silage lactobacillus additive and preparation method thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110591987B (en) * | 2019-11-01 | 2021-05-14 | 四川农业大学 | Lactobacillus salivarius358 and application thereof, silage additive and silage |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0221499A2 (en) * | 1985-11-04 | 1987-05-13 | Microlife Technics, Inc. | Method for inhibiting fungi |
| JPS6416562A (en) * | 1987-07-10 | 1989-01-20 | Kubota Ltd | Production of silage |
| JPH01211487A (en) * | 1988-02-18 | 1989-08-24 | Zenkoku Rakunougiyou Kyodo Kumiai Rengokai | Lactic acid bacteria starter for preparing silage |
| JPH02257875A (en) * | 1989-03-30 | 1990-10-18 | Kubota Ltd | New lactobacillus |
| JPH05336952A (en) * | 1992-06-10 | 1993-12-21 | Valio Mejerien Keskusosuusliik | New microorganism strain, microbial preparation containing the same, use of the microbial strain and preparation for controlling yeast and fungus |
-
1992
- 1992-12-18 JP JP4355857A patent/JPH0757186B2/en not_active Expired - Lifetime
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0221499A2 (en) * | 1985-11-04 | 1987-05-13 | Microlife Technics, Inc. | Method for inhibiting fungi |
| JPS6416562A (en) * | 1987-07-10 | 1989-01-20 | Kubota Ltd | Production of silage |
| JPH01211487A (en) * | 1988-02-18 | 1989-08-24 | Zenkoku Rakunougiyou Kyodo Kumiai Rengokai | Lactic acid bacteria starter for preparing silage |
| JPH02257875A (en) * | 1989-03-30 | 1990-10-18 | Kubota Ltd | New lactobacillus |
| JPH05336952A (en) * | 1992-06-10 | 1993-12-21 | Valio Mejerien Keskusosuusliik | New microorganism strain, microbial preparation containing the same, use of the microbial strain and preparation for controlling yeast and fungus |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003529383A (en) * | 2000-04-07 | 2003-10-07 | ソシエテ デ プロデユイ ネツスル ソシエテ アノニム | Fermented protein hydrolyzate |
| JP2006042647A (en) * | 2004-08-03 | 2006-02-16 | National Agriculture & Bio-Oriented Research Organization | New lactic acid bacterial strain and method for inhibiting silage aerobic deterioration |
| JP2008035728A (en) * | 2006-08-02 | 2008-02-21 | National Agriculture & Food Research Organization | Silage preparation additive, method for preparing silage using the silage preparation additive, and silage preparation kit |
| CN112553103A (en) * | 2020-10-10 | 2021-03-26 | 重庆市畜牧科学院 | Bullwhip grass silage lactobacillus additive and preparation method thereof |
| CN112553103B (en) * | 2020-10-10 | 2022-09-20 | 重庆市畜牧科学院 | Bullwhip grass silage lactobacillus additive and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0757186B2 (en) | 1995-06-21 |
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