JP5940475B2 - Blood cortisol elevation inhibitor - Google Patents
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- JP5940475B2 JP5940475B2 JP2013045366A JP2013045366A JP5940475B2 JP 5940475 B2 JP5940475 B2 JP 5940475B2 JP 2013045366 A JP2013045366 A JP 2013045366A JP 2013045366 A JP2013045366 A JP 2013045366A JP 5940475 B2 JP5940475 B2 JP 5940475B2
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Description
本発明は、反芻動物の血中コルチゾール上昇抑制剤等に関する。詳細には、反芻動物のストレス環境下等における血中コルチゾール上昇を抑制し、及び／又は、反芻動物の飼料摂取量・体重減少を抑制する剤及び方法等に関する。 The present invention relates to a suppressor of blood cortisol elevation in ruminants. More specifically, the present invention relates to an agent and method for suppressing an increase in blood cortisol in a ruminant under a stress environment and / or suppressing a feed intake / weight loss of a ruminant.
コルチゾールは、副腎皮質から分泌される糖質コルチコイドの一種であり、ストレスに敏感に反応して分泌量が増加することから、ストレスホルモンとも呼ばれている。 Cortisol is a kind of glucocorticoid secreted from the adrenal cortex and is also called a stress hormone because it increases the amount of secretion in response to stress.
コルチゾールは炭水化物・脂肪・タンパク質代謝の制御に関わるホルモンであることから、生体には必須のホルモンであるが、過剰なストレスにより多量に分泌された場合、ヒトにおいて脳の海馬を萎縮させることが近年ＰＴＳＤ患者の脳のＭＲＩなどを例として観察されている（非特許文献１）。 Cortisol is a hormone that is involved in the control of carbohydrate, fat, and protein metabolism, so it is an essential hormone in the body. However, in recent years, when it is secreted in large quantities due to excessive stress, it has been known to atrophy the brain hippocampus in humans. MRI of the brain of a PTSD patient has been observed as an example (Non-patent Document 1).
そして、ヒトを含む哺乳動物のストレス予防治療法としては、加水分解コンキオリンやクロレラエキスを用いてストレスホルモン作用を緩和する方法（特許文献１）、生薬成分を用いる方法（特許文献２）などが開示されているが、当業界においては、簡便且つ効率的であり安全性も高いストレス改善ができる成分等の開発が引き続き求められている。これは、反芻動物（特に反芻家畜）を扱う業界においても同じことが言える。 And as a stress prevention treatment method for mammals including humans, a method of reducing stress hormone action using hydrolyzed conchiolin or chlorella extract (Patent Document 1), a method using a crude drug component (Patent Document 2), etc. are disclosed. However, there is a continuing need in the industry for the development of components that can improve stress that are simple, efficient, and highly safe. The same is true in the industry dealing with ruminants (especially ruminant livestock).
一方、乳酸菌発酵物及びプロピオン酸菌発酵物の混合物は、生きている乳酸菌などを摂取させるプロバイオティクスとは異なり、動物自身の腸管内に生息する乳酸菌またはビフィズス菌を増殖させるプレバイオティクス効果を有することが知られており、家畜の体型改良効果やコクシジウム症予防治療効果などを発揮することも知られている（特許文献３、４）。また、プレバイオティクス効果を発揮する素材として、他にもオリゴ糖、酵母細胞壁成分（グルコマンナン）、酵母細胞膜成分（β−グルカン）といったものがある。しかし、いずれも血中コルチゾール濃度や飼料摂取量・体重に対する影響に言及しているものは現段階で見当たらない。 On the other hand, a mixture of fermented lactic acid bacteria and fermented propionic acid bacteria, unlike probiotics that ingest live lactic acid bacteria, has a prebiotic effect to grow lactic acid bacteria or bifidobacteria that live in the intestinal tract of the animal itself. It is also known to have a body shape improving effect and a coccidiosis prevention and treatment effect of domestic animals (Patent Documents 3 and 4). Other materials that exhibit prebiotic effects include oligosaccharides, yeast cell wall components (glucomannan), and yeast cell membrane components (β-glucan). However, none of them mentions effects on blood cortisol concentration, feed intake, or body weight at this stage.
なお、乳酸菌発酵物を有効成分とする抗ストレス剤は、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｈｅｌｖｅｔｉｃｕｓで発酵させた酸乳を有効成分としたもの（特許文献５）、麦汁をラクトバチルス・ペントーサスで発酵させた麦汁発酵物（特許文献６）、ローヤルゼリーを乳酸菌（ラクトバチルス属、ペディオコッカス属、ストレプトコッカス属、ビフィドバクテリウム属）で発酵させた乳酸菌発酵ローヤルゼリー（特許文献７）などが知られている。しかし、乳酸菌発酵物とプロピオン酸菌発酵物の混合物や、Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ、Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉを発酵菌としたものは知られていない。 In addition, the anti-stress agent which uses lactic acid bacteria fermented product as an active ingredient is what used acid milk fermented by Lactobacillus helveticus as an active ingredient (patent document 5), wort fermented product which fermented wort with Lactobacillus pentosas (Patent Document 6), lactic acid bacteria fermented royal jelly (Patent Document 7) obtained by fermenting royal jelly with lactic acid bacteria (Lactobacillus genus, Pediococcus genus, Streptococcus genus, Bifidobacterium genus), and the like are known. However, a mixture of lactic acid bacteria fermented product and propionic acid bacterium fermented product, or Lactobacillus gasseri, Propionibacterium fredenreichii is not known.
また、Ｌ．ｇａｓｓｅｒｉに属する乳酸菌、該乳酸菌含有物に、胃潰瘍の起因であって胃癌との関連も指摘されているＨｅｌｉｃｏｂａｃｔｅｒ ｐｙｌｏｒｉ（ピロリ菌）に対する抑制作用が発見されており（特許文献８）、Ｌ．ｇａｓｓｅｒｉ菌体、その培養物、それから菌体を含む固形物を分離除去した培養物残渣等に、仔ウシをはじめとする仔畜に対する腸内菌そう改善作用、下痢防止作用があることも知られている（特許文献９）が、血中コルチゾール濃度や、飼料摂取量・体重に関する効果については知られていない。 L. An inhibitory action against Helicobacter pylori (H. pylori), which is a cause of gastric ulcer and has been pointed out to be associated with gastric cancer, has been discovered in lactic acid bacteria belonging to gasseri and the lactic acid bacteria-containing substance (Patent Document 8). It is also known that gasseri cells, cultures thereof, and culture residues from which solids containing cells are separated and removed have an intestinal fungus improvement effect and diarrhea prevention effect on calves and other livestock. (Patent Document 9), but the effects on blood cortisol concentration, feed intake and body weight are not known.
さらには、プロピオン酸菌発酵物としては、例えばビフィズス菌増殖促進物質（Ｂｉｆｉｄｏｇｅｎｉｃ Ｇｒｏｗｔｈ Ｓｔｉｍｕｌａｔｏｒ：ＢＧＳ）を産生するＰ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ ＥＴ−３株を１０％ホエー粉還元液で発酵させた、プロピオン酸菌発酵物（Ｐｒｏｆｅｃ）などが知られている。 Furthermore, as a propionic acid bacterium fermented product, for example, P. cerevisiae producing Bifidobacterium Growth Stimulator (BGS) is used. For example, a ferionic acid fermentation product (Profec) obtained by fermenting Fredenreichii ET-3 strain with a 10% whey powder reducing solution is known.
Ｐｒｏｆｅｃ中のＢＧＳの一つに、１，４−ジヒドロキシ−２−ナフトエ酸（１，４−ｄｉｈｙｄｒｏｘｙ−２−ｎａｐｈｔｈｏｉｃ ａｃｉｄ）（ＤＨＮＡ）がある。このＤＨＮＡは、微生物におけるビタミンＫ２（ｍｅｎａｑｕｉｎｏｎｅ）の生合成中間体である。この物質はビフィズス菌のエネルギー代謝過程で生成したＮＡＤＨを効率的に再酸化することにより、増殖を促進する効果が知られている。 One BGS in Profec is 1,4-dihydroxy-2-naphthoic acid (DHNA). This DHNA is a biosynthetic intermediate of vitamin K2 (menaquinone) in microorganisms. This substance is known to have an effect of promoting proliferation by efficiently reoxidizing NADH produced in the energy metabolism process of bifidobacteria.
Ｐｒｏｆｅｃ（ＤＨＮＡ、ＡＣＮＱ：２−ａｍｉｎｏ−３−ｃａｒｂｏｘｙ−１，４−ｎａｐｈｔｈｏｑｕｉｎｏｎｅ）は特定保健用食品として、ビフィズス菌を増やし、腸内環境を改善して、通じを良好に保つ旨の表示許可を得ているが（非特許文献２）、これを実際にウシ等に投与して血中コルチゾールの上昇を抑制する効果を確認した知見はなく、まして、飼料摂取量の低減や体重の低減を抑制することについては、何ら報告例はない。 Profec (DHNA, ACNQ: 2-amino-3-carboxy-1, 4-naphthoquinone) is a food for specified health use, which increases the number of bifidobacteria, improves the intestinal environment, and gives permission to display that it maintains good communication. Although it has been obtained (Non-Patent Document 2), there is no knowledge that this was actually administered to cattle and the like to confirm the effect of suppressing the rise in blood cortisol, let alone the reduction of feed intake and weight reduction There is no report about what to do.
本発明は、天然物由来の有効成分を用いることにより、副作用を伴うことのない安全安心な反芻動物用の血中コルチゾール上昇抑制剤等を提供することを目的としてなされたものである。 The object of the present invention is to provide a safe and reliable blood cortisol rise inhibitor for ruminants and the like that does not cause side effects by using an active ingredient derived from a natural product.
上記目的を達成するため、本発明者らは乳酸菌発酵物及びプロピオン酸菌発酵物が血中コルチゾール濃度、飼料摂取量、体重に与える影響について鋭意検討し、特定の乳酸菌発酵物及びプロピオン酸菌発酵物を有効成分として反芻動物に経口投与することにより、その血中コルチゾール上昇を抑制することに加え、飼料摂取量の減少抑制や体重の減少抑制をすることを見出し、本発明を完成するに至った。 In order to achieve the above-mentioned object, the present inventors diligently studied the effects of fermented lactic acid bacteria and fermented propionic acid bacteria on blood cortisol concentration, feed intake, and body weight, and fermented specific lactic acid bacteria and propionic acid bacteria. In addition to suppressing the rise in blood cortisol by orally administering the product as an active ingredient to ruminants, it has been found that it suppresses the decrease in feed intake and the decrease in body weight, thereby completing the present invention. It was.
すなわち、本発明の実施形態は次のとおりである。
（１）下記する（Ａ）乳酸菌発酵物、及び、（Ｂ）プロピオン酸菌発酵物を有効成分としてなること、を特徴とする反芻動物の血中コルチゾール上昇抑制剤。
（Ａ）乳酸菌発酵物：
ホエー蛋白質誘導体含有培地でラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）に属する乳酸菌を中和培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
（Ｂ）プロピオン酸菌発酵物：
乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地でプロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）に属するプロピオン酸菌を培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
（２）下記する（Ａ）乳酸菌発酵物、及び、（Ｂ）プロピオン酸菌発酵物を有効成分としてなること、を特徴とする反芻動物の血中コルチゾール上昇抑制及び飼料摂取量減少・体重減少抑制剤。
（Ａ）乳酸菌発酵物：
ホエー蛋白質誘導体含有培地でラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）に属する乳酸菌を中和培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
（Ｂ）プロピオン酸菌発酵物：
乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地でプロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）に属するプロピオン酸菌を培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
（３）反芻動物がウシであること、を特徴とする（１）又は（２）に記載の剤。
（４）ウシが出生直後から３２か月齢までのウシであること、を特徴とする（３）に記載の剤。
（５）ウシが出生直後から１２か月齢までの仔ウシであること、を特徴とする（３）に記載の剤。
（６）該乳酸菌発酵物及び／又は該プロピオン酸菌発酵物が、該菌体破砕物及び／又は該菌体培養物を固液分離処理して得た発酵液、その濃縮物、その乾燥物、そのペースト化物、その希釈物、その殺菌物から選ばれる少なくとも１以上であること、を特徴とする（１）〜（５）のいずれか１つに記載の剤。
（７）ホエー蛋白質誘導体が、ホエー蛋白質濃縮物（ＷＰＣ）、ホエー蛋白質分離物（ＷＰＩ）、これらの酵素処理物から選ばれる少なくとも１以上であること、を特徴とする（１）〜（６）のいずれか１つに記載の剤。
（８）乳酸菌がＬａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ ＯＬＬ ２７１６（ＦＥＲＭ ＢＰ−６９９９）であること、を特徴とする（１）〜（７）のいずれか１つに記載の剤。
（９）プロピオン酸菌がＰｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ ＥＴ−３（ＦＥＲＭ ＢＰ−８１１５）株であること、を特徴とする（１）〜（８）のいずれか１項に記載の剤。
（１０）下記する（Ａ）乳酸菌発酵物、及び、（Ｂ）プロピオン酸菌発酵物を離乳期前及び／又は離乳期直後の反芻動物に１０〜５００ｇ／日・個体で経口投与すること、を特徴とする反芻動物の血中コルチゾール上昇を抑制する方法。
（Ａ）乳酸菌発酵物：
ホエー蛋白質誘導体含有培地でラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）に属する乳酸菌を中和培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
（Ｂ）プロピオン酸菌発酵物：
乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地でプロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）に属するプロピオン酸菌を培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
（１１）血中コルチゾール上昇抑制だけでなく、反芻動物の飼料摂取量の減少及び／又は体重減少を併せて抑制すること、を特徴とする（１０）に記載の方法。
That is, the embodiment of the present invention is as follows.
(1) A ruminant blood cortisol rise inhibitor characterized by comprising (A) a fermented lactic acid bacterium and (B) a fermented propionic acid bacterium as active ingredients.
(A) Fermented lactic acid bacteria:
Fermented product prepared by neutralizing lactic acid bacteria belonging to Lactobacillus gasseri in a whey protein derivative-containing medium, its concentrate, its dried product, its pasted product, its liquid product, its diluted product, its sterilized product At least one or more selected from products.
(B) Propionic acid fermented product:
Fermented product prepared by culturing propionic acid bacteria belonging to Propionibacterium fredenreichii in a whey (whey and / or protease-treated product) -containing medium, its concentrate, its dried product, its paste At least one selected from a chemical, a liquid, a dilution, and a sterilized product.
(2) The following (A) lactic acid bacteria fermented product and (B) propionic acid bacteria fermented product are used as active ingredients; Agent.
(A) Fermented lactic acid bacteria:
Fermented product prepared by neutralizing lactic acid bacteria belonging to Lactobacillus gasseri in a whey protein derivative-containing medium, its concentrate, its dried product, its pasted product, its liquid product, its diluted product, its sterilized product At least one or more selected from products.
(B) Propionic acid fermented product:
Fermented product prepared by culturing propionic acid bacteria belonging to Propionibacterium fredenreichii in a whey (whey and / or protease-treated product) -containing medium, its concentrate, its dried product, its paste At least one selected from a chemical, a liquid, a dilution, and a sterilized product.
(3) The agent according to (1) or (2), wherein the ruminant is a cow.
(4) The agent according to (3), wherein the cow is a cow from just after birth to 32 months of age.
(5) The agent according to (3), wherein the cow is a calf from just after birth to 12 months of age.
(6) Fermented liquid obtained by subjecting the fermented lactic acid bacteria and / or fermented propionic acid bacteria to solid-liquid separation of the crushed cells and / or the cultured cell cultures, concentrates thereof, and dried products thereof The agent according to any one of (1) to (5), wherein the agent is at least one selected from a pasted product, a diluted product, and a sterilized product.
(7) The whey protein derivative is at least one or more selected from whey protein concentrate (WPC), whey protein isolate (WPI), and these enzyme-treated products (1) to (6) The agent as described in any one of these.
(8) The agent according to any one of (1) to (7), wherein the lactic acid bacterium is Lactobacillus gasseri OLL 2716 (FERM BP-6999).
(9) The agent according to any one of (1) to (8), wherein the propionic acid bacterium is a Propionibacterium fredenreichii ET-3 (FERM BP-8115) strain.
(10) The following (A) fermented lactic acid bacteria, and (B) fermented propionic acid bacteria are orally administered to ruminants before and / or immediately after the weaning period in an amount of 10 to 500 g / day / individual. A method for suppressing an increase in blood cortisol in a ruminant.
(A) Fermented lactic acid bacteria:
Fermented product prepared by neutralizing lactic acid bacteria belonging to Lactobacillus gasseri in a whey protein derivative-containing medium, its concentrate, its dried product, its pasted product, its liquid product, its diluted product, its sterilized product At least one or more selected from products.
(B) Propionic acid fermented product:
Fermented product prepared by culturing propionic acid bacteria belonging to Propionibacterium fredenreichii in a whey (whey and / or protease-treated product) -containing medium, its concentrate, its dried product, its paste At least one selected from a chemical, a liquid, a dilution, and a sterilized product.
(11) The method according to (10), wherein not only the blood cortisol elevation is suppressed, but also a decrease in feed intake and / or weight loss of ruminants is suppressed.
（１２）下記する（Ａ）乳酸菌発酵物、及び、（Ｂ）プロピオン酸菌発酵物を有効成分としてなること、を特徴とする反芻動物の飼料摂取量減少及び／又は体重減少抑制剤。
（Ａ）乳酸菌発酵物：
ホエー蛋白質誘導体含有培地でラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）に属する乳酸菌を中和培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
（Ｂ）プロピオン酸菌発酵物：
乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地でプロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）に属するプロピオン酸菌を培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上。
(12) A feed intake decrease and / or weight loss inhibitor for ruminants, characterized in that (A) a fermented product of lactic acid bacteria and (B) a fermented product of propionic acid bacteria are used as active ingredients.
(A) Fermented lactic acid bacteria:
Fermented product prepared by neutralizing lactic acid bacteria belonging to Lactobacillus gasseri in a whey protein derivative-containing medium, its concentrate, its dried product, its pasted product, its liquid product, its diluted product, its sterilized product At least one or more selected from products.
(B) Propionic acid fermented product:
Fermented product prepared by culturing propionic acid bacteria belonging to Propionibacterium fredenreichii in a whey (whey and / or protease-treated product) -containing medium, its concentrate, its dried product, its paste At least one selected from a chemical, a liquid, a dilution, and a sterilized product.
本発明によれば、特定の乳酸菌発酵物及びプロピオン酸菌発酵物（プロピオン酸菌による乳清発酵物）を有効成分とする剤を反芻動物に経口投与するだけで、ストレス環境下等における反芻動物の血中コルチゾール濃度上昇を抑制することができ、輸送などによるストレスを軽減させることができる。さらに、反芻動物の飼料摂取量減少・体重減少も併せて抑制することができる。本発明は、出生直後から３２か月齢までの個体（特にウシ）に対して極めて有効である。 According to the present invention, a ruminant under a stress environment or the like can be obtained by simply orally administering to a ruminant an agent containing a specific lactic acid bacterium fermented product and a propionic acid bacterium fermented product (whey fermented product of propionic acid bacterium) as an active ingredient. The increase in blood cortisol concentration can be suppressed, and the stress due to transportation and the like can be reduced. Furthermore, it is possible to suppress ruminant animal feed intake reduction and weight loss. The present invention is extremely effective for an individual (particularly a cow) from just after birth to 32 months of age.
以下、本発明を詳細に説明する。 Hereinafter, the present invention will be described in detail.
本発明においては、ホエー蛋白質誘導体含有培地でラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）に属する乳酸菌を中和培養して調製した発酵物、及び／又は、その発酵物の濃縮物、乾燥物（噴霧乾燥物、凍結乾燥物、真空乾燥物、ドラム乾燥物の少なくともひとつ）、ペースト化物、液状物、希釈物、殺菌物から選ばれる少なくとも１以上（以下、これらを総称して乳酸菌発酵物と言う場合もある）と、乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地でプロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）に属するプロピオン酸菌を培養して調製した発酵物、及び／又は、その発酵物の濃縮物、乾燥物（噴霧乾燥物、凍結乾燥物、真空乾燥物、ドラム乾燥物の少なくともひとつ）、ペースト化物、液状物、希釈物、殺菌物から選ばれる少なくとも１以上（以下、これらをプロピオン酸菌発酵物あるいはプロピオン酸菌による乳清発酵物と言う場合もある）の両方を反芻動物の血中コルチゾール上昇抑制剤等の有効成分として使用する。 In the present invention, a fermented product prepared by neutralizing and cultivating lactic acid bacteria belonging to Lactobacillus gasseri in a whey protein derivative-containing medium, and / or a concentrate or a dried product (spray-dried product) of the fermented product , Freeze-dried product, vacuum-dried product, drum-dried product), pasted product, liquid product, diluted product, sterilized product or more (hereinafter collectively referred to as lactic acid bacteria fermented product) And a fermented product prepared by culturing propionic acid bacteria belonging to Propionibacterium fredenreichii in a whey (whey and / or protease-treated product) -containing medium and / or a fermented product thereof Concentrates, dried products (spray dried products, freeze dried products, At least one selected from a vacuum-dried product, a drum-dried product, a paste product, a liquid product, a diluted product, and a sterilized product (hereinafter referred to as a propionic acid bacterium fermented product or a propionic acid bacterium fermented whey product) In some cases, both are used as active ingredients such as blood cortisol elevation inhibitors in ruminants.
乳酸菌発酵物、プロピオン酸菌発酵物の含有成分としては、Ｌ．ｇａｓｓｅｒｉ又はＰ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉを培養して得た発酵物（菌体、培地成分、代謝分泌物等を含む培養物全体）、それから分離した菌体の破砕物、該発酵物（菌体培養物）から菌体を含む固形物を固液分離処理により除去した発酵液（培養物残渣；培養液の混濁した部分ではなく澄んだ部分である培養上澄液を包含する）から選ばれる少なくとも１以上が挙げられる。本発明においては、これら発酵物中にＬ．ｇａｓｓｅｒｉ、Ｐ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉが生菌として存在する必要がない（菌体成分、菌体分泌物から選ばれる１以上のみであっても充分な効果を奏する）ことが特徴である。 Examples of the components contained in the fermented lactic acid bacteria and fermented propionic acid bacteria include L. gasseri or P.I. Fermented product obtained by culturing Fredenreichii (whole cell culture medium, culture medium components, metabolic secretions, etc.), crushed cell isolated from the fermented product (fungal cell culture) Examples thereof include at least one or more selected from fermentation broths obtained by removing solids by solid-liquid separation treatment (culture residue; including culture supernatant that is not a turbid part of the culture liquid but a clear part). In the present invention, L. gasseri, P.M. It is a feature that Fredenreichii does not need to exist as a living bacterium (there is sufficient effect even if only one or more selected from microbial components and microbial secretions).
本発明の乳酸菌発酵物を調製する際に用いる発酵菌は、ラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）に属する乳酸菌である。特に、ラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）ＯＬＬ ２７１６株を用いることが好ましい。
なお、ラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）ＯＬＬ ２７１６株は、独立行政法人製品評価技術基盤機構・特許生物寄託センター（日本国〒３０５−８５６６茨城県つくば市東１−１−１つくばセンター中央第６）に、２０００年（平成１２年）１月１４日付けで国際寄託されており、その受託番号はＦＥＲＭ ＢＰ−６９９９である。
The fermenting bacteria used when preparing the fermented lactic acid bacteria of the present invention are lactic acid bacteria belonging to Lactobacillus gasseri. In particular, Lactobacillus gasseri OLL 2716 strain is preferably used.
The Lactobacillus gasseri OLL 2716 strain is the National Institute for Product Evaluation Technology and Patent Biological Deposit Center (Tokuba Center 1-1-1 Tsukuba City East, Tsukuba City, Japan 305-8586) In addition, an international deposit was made on January 14, 2000, and the deposit number is FERM BP-6999.
Ｌ．ｇａｓｓｅｒｉの培養方法は、ｐＨを一定範囲（例えばｐＨ４．０〜８．０）に保持して所定菌数に達するまで（例えば２０〜４０℃、好ましくは３５〜３８℃で２４〜４８時間程度）の中和培養を行う。この高濃度培養法では、菌体および代謝産物の多い培養物が得られる。その他の培養条件は、常法に従えば良い。 L. The gasseri culture method keeps the pH within a certain range (for example, pH 4.0 to 8.0) and reaches a predetermined number of bacteria (for example, 20 to 40 ° C., preferably 35 to 38 ° C. for about 24 to 48 hours). Perform neutralization culture. In this high concentration culture method, a culture rich in bacterial cells and metabolites can be obtained. Other culture conditions may be according to ordinary methods.
培地としては、ホエー蛋白質誘導体含有培地を使用するが、好適なＬ．ｇａｓｓｅｒｉの高濃度培養液は、ホエー蛋白質誘導体及び乳糖を含有する培地、特にホエー蛋白質誘導体及び乳糖を主成分とする培地を使用し、ｐＨを４．５〜７．０の範囲に保持して、菌数が１０８〜１０１１、好ましくは１０９から１０１０となるまで中和培養して得られる。なお、通常の培養方法の場合では、培養物中の菌数は１０９以下にしかならない。 As the medium, a whey protein derivative-containing medium is used. gasseri high-concentration culture medium uses a medium containing whey protein derivative and lactose, particularly a medium mainly composed of whey protein derivative and lactose, and maintains the pH in the range of 4.5 to 7.0, It is obtained by neutralization culture until the number of bacteria becomes 10 8 to 10 11 , preferably 10 9 to 10 10 . In the case of a normal culture method, the number of bacteria in the culture is only 10 9 or less.
ホエー蛋白質誘導体としては、ホエー蛋白質濃縮物（Ｗｈｅｙ Ｐｒｏｔｅｉｎ Ｃｏｎｃｅｎｔｒａｔｅ、ＷＰＣ）、ホエー蛋白質分離物（Ｗｈｅｙ Ｐｒｏｔｅｉｎ Ｉｓｏｌａｔｅ、ＷＰＩ）、これらの加水分解物から選ばれる少なくともひとつが使用され、市販品も適宜使用可能である。 As the whey protein derivative, at least one selected from whey protein concentrate (Whey Protein Concentrate, WPC), whey protein isolate (Whey Protein Isolate, WPI), and a hydrolyzate thereof can be used, and commercially available products can also be used as appropriate. It is.
ＷＰＣは、チーズやカゼインを製造する際に生じるホエーを限外濾過・ゲル濾過・乳糖結晶分離などの方法で処理し、蛋白質含量を通常３５〜８５％（固形分換算）まで高めたものである。ＷＰＩは、ＷＰＣと区別されるものであって、イオン交換法などの方法で蛋白質含量を９５％（固形分換算）程度まで高めたものである。 WPC is obtained by treating whey produced in the production of cheese and casein by methods such as ultrafiltration, gel filtration, and lactose crystal separation, and increasing the protein content to 35-85% (in terms of solid content). . WPI is distinguished from WPC and has a protein content increased to about 95% (in terms of solid content) by a method such as an ion exchange method.
加水分解物としては、ＷＰＣ、ＷＰＩ、ホエー、ホエー蛋白質から選ばれる少なくともひとつを、ペプシン、トリプシン、パパイン等の蛋白質分解酵素等の酵素で処理した酵素処理物が使用される。 As the hydrolyzate, an enzyme-treated product obtained by treating at least one selected from WPC, WPI, whey, and whey protein with an enzyme such as a proteolytic enzyme such as pepsin, trypsin, or papain is used.
乳酸菌発酵物は、菌体成分及び／又は代謝成分（有機酸、ペプチド、オリゴ糖など）による生理作用を有し、また、Ｌ．ｇａｓｓｅｒｉは、乳酸やバクテリオシンを産生するだけでなく、オリゴＤＮＡも産生し、これら産生物質の生理作用などが有機的に働いて本発明の効果を奏するものと、現時点では考えられている。 The lactic acid bacteria fermented product has a physiological effect due to bacterial cell components and / or metabolic components (organic acids, peptides, oligosaccharides, etc.). Gasseri is considered not only to produce lactic acid and bacteriocin but also to produce oligo DNA, and physiological effects of these produced substances work organically to achieve the effects of the present invention.
本発明に係る乳酸菌発酵物は、Ｌ．ｇａｓｓｅｒｉ菌体由来成分、その他のＬ．ｇａｓｓｅｒｉ由来物（代謝成分、分泌成分など）だけからなり、これをそのまま、あるいはこれに賦形剤（デンプン、デキストリン、乳成分、ケイ酸等）、飼料成分、飲料水等を添加したものも包含する。本発酵物は、液状、ペースト状、固体状のいずれの状態にもすることができる。 The lactic acid bacteria fermented product according to the present invention is obtained from L. gasseri cells-derived components, other L. It consists only of gasseri-derived products (metabolic components, secretory components, etc.) and includes those as they are or with excipients (starch, dextrin, milk components, silicic acid, etc.), feed ingredients, drinking water, etc. To do. The present fermented product can be in any state of liquid, paste or solid.
一方、Ｐ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉによる乳清発酵物（Ｐｒｏｆｅｃ等）は、ヒトの腸内のビフィズス菌（ビフィドバクテリウム：Ｂｉｆｉｄｏｂａｃｔｅｒｉｕｍ）を特異的に増やすことから、特定保健用食品の関与成分として認可され、現在、実際に利用されている。 On the other hand, P.I. The fermented whey by Fredenreichii (Profec, etc.) is specifically approved as a component to be involved in foods for specific health use because it specifically increases Bifidobacterium in the human intestine. It's being used.
プロピオン酸菌による乳清発酵物としては、上記Ｐｒｏｆｅｃが使用できるほか、Ｐ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉに属するプロピオン酸菌を乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地で培養し、得られた発酵物、単離した菌体の破砕物、培養液（培養上清）などを広く使用することができる。 Profec can be used as a whey fermented product by propionic acid bacteria. culturing propionic acid bacteria belonging to Fredenreichii in a medium containing whey (whey and / or its protease-treated product), and widely obtaining fermented products, isolated crushed cells, culture fluid (culture supernatant), etc. Can be used.
Ｐ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉに属するプロピオン酸菌としては、ＢＧＳ産生菌が適宜使用され、その非限定例としては、Ｐ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ ＡＴＣＣ ６２０７、同ＡＴＣＣ ８２６２、同ＩＦＯ １２４２４、同ＩＦＯ １２４２６、同ＩＦＯ １２３９１が例示され、また、Ｐｒｏｆｅｃに使用されているＥＴ−３株も入手、使用可能である。
なお、プロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）ＥＴ−３株は、独立行政法人製品評価技術基盤機構・特許生物寄託センター（日本国〒３０５−８５６６茨城県つくば市東１−１−１つくばセンター中央第６）に、２００２年（平成１４年）７月１１日付けで国際寄託されており、その受託番号はＦＥＲＭ ＢＰ−８１１５である。
P. As propionic acid bacteria belonging to Freudrenreichii, BGS-producing bacteria are appropriately used. Fredendenichii ATCC 6207, ATCC 8262, IFO 12424, IFO 12426, and IFO 12391 are exemplified, and the ET-3 strain used in Profec is also available and usable.
Propionibacterium fredenreichichi ET-3 is an independent administrative agency of the National Institute for Product Evaluation Technology and Patent Biological Deposit Center (Tsukuba Center 1-1-1 Tsukuba City, Ibaraki Prefecture 305-8586) In the middle 6), the international deposit was made on July 11, 2002, and the deposit number is FERM BP-8115.
Ｐ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉの培養は、乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地を使用すること（好ましくは１〜２０％、更に好ましくは５〜１５％）以外は、常法にしたがって好気的又は嫌気的（例えば、窒素ガス加圧下（０．５ｋｇ／ｃｍ２））培養によればよい。通常、２０〜４０℃、ｐＨ６〜８程度で液体培養すればよい。 P. Fredenreichii culture is aerobic according to a conventional method, except that a medium containing whey (whey and / or its protease-treated product) is used (preferably 1 to 20%, more preferably 5 to 15%). Anaerobic (for example, under nitrogen gas pressure (0.5 kg / cm 2 )) culture may be used. Usually, liquid culture may be performed at 20 to 40 ° C. and pH of 6 to 8.
なお、このようにして調製したＰ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉによる乳清発酵物も、Ｐ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ菌体由来成分、その他のＰ．ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ由来物（代謝成分、分泌成分など）だけからなり、そのまま、あるいは賦形剤（脱脂粉乳、デンプン、穀粉、糖類等）、飼料成分、飲料水等を添加したものも包含する。本発酵物も、液状、ペースト状、固体状のいずれの状態にもすることができる。 In addition, the P.P. The whey fermented product by Freudrenreichii is also disclosed in P. Fredenreichii cell-derived component and other P. cerevisiae cells. It includes only those derived from fredenreichii (metabolic components, secretory components, etc.), and those to which excipients (fat dry milk, starch, flour, saccharides, etc.), feed ingredients, drinking water, etc. are added as they are. The present fermented product can also be in a liquid, pasty, or solid state.
本発明においては、通常、上記した乳酸菌発酵物とプロピオン酸菌発酵物を併用して用いるが、これらの発酵物は別々に使用してもよいし、両発酵物の混合物（共存物）を使用してもよい。乳酸菌発酵物とプロピオン酸菌発酵物の混合比は、１：１０〜１００：１が好ましく、１：５〜５０：１（乾物重量比）が更に好ましい。 In the present invention, the above-mentioned fermented lactic acid bacteria and fermented propionic acid bacteria are usually used in combination, but these fermented products may be used separately or a mixture of both fermented products (coexisting product) is used. May be. The mixing ratio of the lactic acid bacterium fermented product and the propionic acid bacterium fermented product is preferably 1:10 to 100: 1, and more preferably 1: 5 to 50: 1 (dry matter weight ratio).
なお、両発酵物の混合物は、これを有効成分として組成物とすることができる。当該組成物は、そのまま、あるいは賦形剤や飼料成分と混合して飼料として、反芻動物に経口にて投与・給与される。例えば、飲料水に混ぜたり、代用乳に混合したりすることができる。その投与量は後記するようにそれほど多くなく、反芻動物に負担がかからない点でも本発明は極めて特徴的である。 In addition, the mixture of both fermented products can be made into a composition by using this as an active ingredient. The composition is administered or fed to ruminants orally as it is or as a feed mixed with excipients or feed ingredients. For example, it can be mixed with drinking water or mixed with milk substitute. The dose is not so large as described later, and the present invention is very characteristic in that the ruminant is not burdened.
これら両発酵物の混合物としては、市販品も使用することができる。市販品としては、例えばラクトバチルス・ガッセリＯＬＬ ２７１６株（ＦＥＲＭ ＢＰ−６９９９）発酵物とＰｒｏｆｅｃ（プロフェック：株式会社明治 商品名）の混合物を含有するプレビオサポート（明治飼糧株式会社登録商標、ＰＳ）が使用できる。 A commercial product can also be used as a mixture of these two fermented products. As a commercially available product, for example, Prebio Support (Meiji Kayaku Co., Ltd., registered trademark, PS) containing a mixture of Lactobacillus gasseri OLL 2716 strain (FERM BP-6999) fermented product and Profec (Profec: Meiji brand name) Can be used.
本有効成分の経口投与量としては、動物の体重１ｋｇあたり両発酵物の混合物として０．１〜５．０ｇ／日（好ましくは、動物の体重１ｋｇあたり０．４〜２．０ｇ／日）で、特に、出生直後〜３２か月齢までの個体に経口投与又は給与するのが好ましい。経口投与量が当該範囲より多い場合には、その効果という点において特段の問題はないが、当該範囲より少ない場合には、本発明の効果が十分発揮されない恐れがあるため好ましくない。例えば、仔ウシ（体重５０〜１００ｋｇ程度）を例とすると、１日あたり１０〜５００ｇ／頭、好ましくは２０〜１００ｇ／頭が好適な経口投与量として示される。 The oral dose of the active ingredient is 0.1 to 5.0 g / day (preferably 0.4 to 2.0 g / day of animal body weight) as a mixture of both fermented products per kg of animal body weight. In particular, it is preferable to orally administer or feed to an individual immediately after birth to 32 months of age. When the oral dose is larger than the above range, there is no particular problem in terms of the effect. However, when the oral dose is less than the above range, the effect of the present invention may not be sufficiently exhibited, which is not preferable. For example, taking calf (weight of about 50 to 100 kg) as an example, 10 to 500 g / head, preferably 20 to 100 g / head per day is shown as a suitable oral dose.
本発明の対象動物は、反芻動物である。特に、その乳あるいは乳加工品が食用となる反芻家畜が好適例として示され、例えば、牛（特にホルスタイン種、ジャージー種などの乳牛）、羊、山羊などであるが、これに限定されるものではない。その他、ラクダ、ラマなども例示され、特に、出生直後から３２か月齢程度の個体、好ましくは出生直後から１２か月齢程度の個体（仔畜）、更に好ましくは４日齢から１２０日齢程度の個体を対象とするのが好適である。 The target animal of the present invention is a ruminant. In particular, ruminant livestock whose milk or dairy products are edible are shown as preferred examples, such as cows (particularly dairy cows such as Holstein and Jersey), sheep, goats, etc., but are not limited thereto. is not. In addition, camels and llamas are also exemplified, and in particular, an individual of about 32 months of age immediately after birth, preferably an individual of about 12 months of age after birth (offspring), more preferably about 4 to 120 days of age. It is preferable to target an individual.
以上述べてきたように、本発明は、乳酸菌発酵物及びプロピオン酸菌発酵物の有用性に関して更に研究を重ねた結果、遂に完成されたものであって、乳酸菌培養物およびプロピオン酸菌発酵物の両方を有効成分とし、ストレス環境下等における反芻動物の血中コルチゾール上昇抑制及び飼料摂取量減少抑制・体重減少抑制をする技術を基本的技術思想とするものである。 As described above, the present invention was finally completed as a result of further research on the usefulness of lactic acid bacteria fermented products and propionic acid bacterium fermented products. The basic technical idea is to use both as active ingredients, and to suppress the increase of blood cortisol in ruminant animals under the stress environment and the like, and suppress the decrease in feed intake and body weight.
以下、本発明の実施例について述べるが、本発明はこれらの実施例のみに限定されるものではなく、本発明の技術的思想内においてこれらの様々な変形が可能である。 Examples of the present invention will be described below, but the present invention is not limited to these examples, and various modifications can be made within the technical idea of the present invention.
ホルスタイン種新生仔ウシ（１４頭）に対し、出生直後から３日齢までＩｇＧを強化した人工初乳を与えた。４日齢より仔ウシを２群に分け、４２日齢に達するまで試験群（７頭）に乳酸菌発酵物及びプロピオン酸菌による乳清発酵物の混合物であるプレビオサポート（２０ｇ／日）を含有した代用乳を、対照群（７頭）に代用乳のみを給与した。試験期間中は、全ての動物に仔ウシ用配合飼料（スターター）を自由摂取させた。
４２日齢時において各群で健康状態の良い動物を６頭ずつ選抜して車両に乗せ、一般道１時間、高速道路３時間輸送し、輸送前後における血中のコルチゾール濃度とスターター摂取量を調査した。
Holstein newborn calves (14 heads) were given artificial colostrum enriched with IgG from just after birth until 3 days of age. Calves were divided into 2 groups from 4 days of age, and the test group (7 animals) contained prebiosupport (20 g / day), which was a mixture of lactic acid bacteria fermentation products and propionate bacteria whey fermentation products until 42 days of age were reached. The substitute milk was fed only to the control group (7 animals). During the test period, all animals were allowed to freely ingest the calf formula feed (starter).
At the age of 42 days, 6 animals of good health in each group were selected and placed in a vehicle, transported for 1 hour on the general road and 3 hours on the highway, and the blood cortisol concentration and starter intake before and after transport were investigated. did.
なお、人工初乳は、ＩｇＧ含量が２６．７％（６０ｇ／２２５ｇ）のバイエルメディカル株式会社製品（商品名：ヘッドスタート、成分値：蛋白質４５％）を使用した。更に、代用乳は乳蛋白および植物性油脂を主原料とした明治飼糧株式会社製品（商品名：プレミアムメイラック、成分値：蛋白質２４％、脂肪２０％）を使用した。仔ウシ用配合飼料は、とうもろこしおよび大豆油粕を主原料とした明治飼糧株式会社製品（商品名：ミルキースターターＰＦ、成分値：蛋白質２５％、可消化養分総量７６％）を使用した。 The artificial colostrum used was a Bayer Medical Co., Ltd. product (trade name: head start, component value: protein 45%) with an IgG content of 26.7% (60 g / 225 g). In addition, milk substitute and Meiji Feed Co., Ltd. products (trade name: Premium Mayrak, component value: protein 24%, fat 20%) using milk protein and vegetable oil as main raw materials were used. The compound feed for calf was Meiji Feed Co., Ltd. product (trade name: Milky Starter PF, component value: 25% protein, total digestible nutrient amount 76%) using corn and soybean meal as the main raw materials.
試験の結果、試験群において輸送直後（輸送０時間）、輸送１日後、輸送２日後の血中コルチゾール濃度が対照群より低く抑えられていることが明らかとなった（図１）。また、スターター摂取量は、輸送１日後以降では対照群と比較して試験群に於いて有意に高くなった（図２）。従って、乳酸菌発酵物及びプロピオン酸菌による乳清発酵物の両方を哺乳中の期間（３８日間）に経口投与することにより、輸送後における血中コルチゾール濃度の上昇が抑制されると同時にスターターの摂取量が上昇し、輸送による影響を極めて少なくすることが示された。 As a result of the test, it was revealed that the blood cortisol concentration in the test group was suppressed to be lower than that in the control group immediately after transport (transport 0 hour), 1 day after transport, and 2 days after transport (FIG. 1). In addition, the starter intake was significantly higher in the test group than in the control group after 1 day after transportation (FIG. 2). Therefore, by administering orally both fermented lactic acid bacteria and fermented whey by propionic acid bacteria during the feeding period (38 days), the rise in blood cortisol concentration after transportation is suppressed and intake of the starter The amount was increased and shown to have very little impact from transportation.
離乳直後（４２日齢）のホルスタイン種仔ウシ（１２頭）を２群に分け、試験群（６頭）に乳酸菌発酵物及びプロピオン酸菌による乳清発酵物の混合物であるプレビオサポート（１００ｇ／回）を含有した代用乳を２回給与し、対照群（６頭）には代用乳のみを２回給与した。この間、全ての動物に仔ウシ用配合飼料（スターター）を自由摂取させた。
給与終了後、全ての仔ウシを車両に乗せ、一般道１時間、高速道路３時間輸送し、輸送前後における血中コルチゾール濃度と体重を調査した。
Immediately after weaning (42 days of age), Holstein calves (12 heads) were divided into two groups, and the test group (6 heads) was prebiosupport (100 g / 2), and the control group (6 animals) received only the milk substitute twice. During this time, all the animals were allowed to ingest the calf formula feed (starter) freely.
After the end of the salary, all calves were placed in a vehicle and transported for 1 hour on a general road and 3 hours on a highway, and blood cortisol concentration and body weight before and after transport were examined.
なお、代用乳は乳蛋白および植物性油脂を主原料とした明治飼糧株式会社製品（商品名：プレミアムメイラック、成分値：蛋白質２４％、脂肪２０％）を使用した。仔ウシ用配合飼料は、とうもろこしおよび大豆油粕を主原料とした明治飼糧株式会社製品（商品名：ミルキースターターＰＦ、成分値：蛋白質２５％、可消化養分総量７６％）を使用した。 The milk replacer used was a product of Meiji Feed Co., Ltd. (trade name: Premium Mayrak, component value: 24% protein, 20% fat) using milk protein and vegetable oil as main ingredients. The compound feed for calf was Meiji Feed Co., Ltd. product (trade name: Milky Starter PF, component value: 25% protein, total digestible nutrient amount 76%) using corn and soybean meal as the main raw materials.
試験の結果、対照群において輸送直後における体重の減少率が平均５．５％であったのに対し、試験群では平均３．１％と低く抑えられた（図３）。また、輸送後における血中コルチゾール濃度は対照群と比較して試験群に於いて低く抑えられた（図４）。従って、乳酸菌発酵物及びプロピオン酸菌による乳清発酵物の両方を輸送前のわずか２回の経口投与により、輸送後における体重の減少が抑制され、更に血中コルチゾール濃度の上昇が抑制された。 As a result of the test, the average decrease in body weight immediately after transport in the control group was 5.5%, whereas in the test group, the average decrease was 3.1% (FIG. 3). Moreover, the blood cortisol concentration after transportation was suppressed to a low level in the test group as compared with the control group (FIG. 4). Therefore, the oral administration of both the lactic acid bacteria fermented product and the propionate-based whey fermented product before oral transportation only suppressed the decrease in body weight after transportation and further suppressed the increase in blood cortisol concentration.
以上より、本発明に係る乳酸菌発酵物及びプロピオン酸菌による乳清発酵物の混合物を４日齢から４２日齢程度の仔ウシに経口投与することにより、車両輸送時及び輸送後の血中コルチゾール上昇を抑制し、且つ、スターター摂取量減少、体重減少も抑制し、その輸送ストレスを軽減することが示された。 As mentioned above, blood cortisol at the time of vehicle transportation and after transportation by orally administering the mixture of the fermented lactic acid bacteria according to the present invention and the fermented whey produced by propionic acid bacteria to calves of about 4 to 42 days of age. It was shown that it suppresses the rise, suppresses starter intake and weight loss, and reduces the transport stress.
なお、本発明を要約すれば次のとおりである。 The present invention is summarized as follows.
すなわち、本発明は、天然物由来の有効成分を用いることにより、副作用を伴うことのない安全安心な反芻動物用の血中コルチゾール上昇抑制剤等を提供することを目的とする。 That is, an object of the present invention is to provide a safe and reliable blood cortisol increase inhibitor for ruminants and the like that does not involve side effects by using an active ingredient derived from a natural product.
そして、ホエー蛋白質誘導体含有培地でラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）に属する乳酸菌を中和培養して調製した乳酸菌発酵物、及び、乳清（ホエー及び／又はそのプロテアーゼ処理物）含有培地でプロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）に属するプロピオン酸菌を培養して調製したプロピオン酸菌発酵物を有効成分とすることにより、反芻動物用の血中コルチゾール上昇抑制剤等を提供することができる。 Then, a fermented lactic acid bacterium prepared by neutralizing lactic acid bacteria belonging to Lactobacillus gasseri in a whey protein derivative-containing medium, and a whey (whey and / or its protease-treated product) -containing propioni By using as an active ingredient a propionic acid bacteria fermented product prepared by culturing propionic acid bacteria belonging to Propionibacterium fredenreichii, it is possible to provide a blood cortisol rise inhibitor for ruminants and the like. .
本発明において寄託されている微生物の受託番号を下記に示す。
（１）ラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）ＯＬＬ ２７１６株（ＦＥＲＭ ＢＰ−６９９９）。
（２）プロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）ＥＴ−３株（ＦＥＲＭ ＢＰ−８１１５）。
The accession numbers of the microorganisms deposited in the present invention are shown below.
(1) Lactobacillus gasseri OLL 2716 strain (FERM BP-6999).
(2) Propionibacterium fredenreichii ET-3 strain (FERM BP-8115).
Claims (4)
（Ａ）乳酸菌発酵物：
ホエー蛋白質誘導体含有培地でラクトバチルス・ガッセリ（Ｌａｃｔｏｂａｃｉｌｌｕｓ ｇａｓｓｅｒｉ）ＯＬＬ ２７１６（ＦＥＲＭ ＢＰ−６９９９）株を中和培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上を得る。
（Ｂ）プロピオン酸菌発酵物：
ホエー及び／又はそのプロテアーゼ処理物含有培地でプロピオニバクテリウム・フロイデンライヒ（Ｐｒｏｐｉｏｎｉｂａｃｔｅｒｉｕｍ ｆｒｅｕｄｅｎｒｅｉｃｈｉｉ）ＥＴ−３（ＦＥＲＭ ＢＰ−８１１５）株を培養して調製した発酵物、その濃縮物、その乾燥物、そのペースト化物、その液状物、その希釈物、その殺菌物から選ばれる少なくとも１以上を得る。 10 to 500 g / day / individual amount of (A) fermented lactic acid bacteria and (B) fermented propionic acid bacteria obtained by the following preparation method for ruminants from 4 days to 120 days before vehicle transportation A method of suppressing an increase in blood cortisol in ruminants during and after transportation of the vehicle, and at the same time, suppressing a decrease in feed intake and weight loss.
(A) Fermented lactic acid bacteria:
A fermentation product prepared by neutralizing Lactobacillus gasseri OLL 2716 (FERM BP-6999) strain in a whey protein derivative-containing medium, a concentrate thereof, a dried product thereof, a pasted product thereof, a liquid product thereof, At least one selected from the diluted product and the sterilized product is obtained .
(B) Propionic acid fermented product:
Fermented product prepared by culturing Propionibacterium fredenreich ET-3 (FERM BP-8115) strain in a medium containing whey and / or its protease-treated product, its concentrate, its dried product, its At least one or more selected from pasted material, liquid material, diluted material, and sterilized material is obtained .
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