JP2004321068A

JP2004321068A - Lipid metabolism-promoting meal and feed additive

Info

Publication number: JP2004321068A
Application number: JP2003119791A
Authority: JP
Inventors: Kenji Sakai; 謙二酒井; Miki Umeki; 美樹梅木; Kazutoshi Ogami; 和敏大上; Satoshi Mochizuki; 聡望月
Original assignee: OITA TECHNOLOGY LICENSING ORGA; OITA TECHNOLOGY LICENSING ORGANIZATION Ltd
Current assignee: OITA TECHNOLOGY LICENSING ORGA; OITA TECHNOLOGY LICENSING ORGANIZATION Ltd
Priority date: 2003-04-24
Filing date: 2003-04-24
Publication date: 2004-11-18

Abstract

<P>PROBLEM TO BE SOLVED: To provide a lipid metabolism-promoting meal or feed additive solving the problems that (1) a diarrheal disease is caused by excessively administering water-soluble dietary fibers and (2) there have hitherto been no reports on oligosaccharides or microorganismal preparations enabling metabolism promotion regarding all of various lipids in the serum and liver, etc. <P>SOLUTION: The lipid metabolism-promoting meal/feed additive contains Lactobacillus rhamnosus and cellobiose in the weight ratio of 1:(2.5-8), having (a) the effect of normalizing the intestine( shortening the passage time of contents in the intestine and excreta water content holding ability in humans and animals ) and the effect of ameliorating diarrheal diseases often caused by water-soluble dietary fibers and (b) the effect of lowering body total lipid level in the serum or liver( lipid metabolism-promoting effect ). <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

【０００１】
【発明の属する技術分野】
本発明は、乳酸菌菌体と水溶性食物繊維を同時に含有させることで，整腸効果と共に、各々単独の場合に比べて著しい脂質濃度低下効果を示す脂質代謝促進用肥料食・飼料添加物に関するものである。
【０００２】
【従来の技術】
従来から乳酸菌・発酵乳の機能に関心がもたれるきっかけとなる報告は，二十世紀初頭にメチニコフによって提唱された不老長寿説といわれる。これは発酵乳を常食している人々の寿命は長く、それは乳酸菌が腸内に定着し有害菌による腐敗を抑えるためであるとの考えであった。
【０００３】
近年では，プロバイオティクス・バイオジェニックスの観点からも大きく注目されるようになり、機能性食品成分としての乳酸菌に関する多くの研究がなされている。
【０００４】
これまでに機能性に関する報告として、整腸作用，血中コレステロール低下作用、抗腫瘍作用、免疫賦活作用、血圧低下作用などが知られている。しかしこれらはすべて乳酸菌単独で摂取した例である。
一方、ある種の水溶性食物性繊維が整腸、腸内腐敗抑制、大腸ガン抑制等の効果を示すことも広く知られている。
【０００５】
【発明が解決しようとする課題】
しかしながら、水溶性食物繊維は、過剰の投与によって下痢症状を引き起こす問題点があった。
また血清や肝臓などにおいて各種脂質すべてに関して代謝促進を促すことのできるオリゴ糖、あるいは微生物製剤に関する報告はこれまでなかった。
【０００６】
【課題を解決するための手段】
本発明は、前記課題を解決した優れたものであり、その特徴とするところは、乳酸菌（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｈａｍｎｏｓｕｓ）菌体とセロビオースを配合比で、１：２．５〜１０含有せしめたことを特徴とする整腸及び脂質代謝促進用肥料食・飼料添加物にある。
【０００７】
【発明の実施の形態】
即ち本発明者等は、分離、同定した乳酸菌Ｌａｃｔｏｂａｃｉｌｌｕｓｒｈａｍｎｏｓｕｓ（以下単に乳酸菌と称する）について、その機能性の発現を期待して、ラットに投与し脂質代謝に及ぼす影響を検討した結果，乳酸菌の単独投与では，血清脂質に対する影響は認められなかったが、乳酸菌をセロビオースと同時に投与すると，血清中の総脂質，中性脂肪，β−リポタンパクおよびコレステロール濃度が低下すると供に、肝臓脂質についても減少すると言う相乗効果の新知見を得て本発明を完成したものである。
【０００８】
本発明において、前記乳酸菌体とセロビオースとの配合比は、１：２．５〜８にすることによりｐＨの低下と共に酢酸及びプロピオン酸量が有意に増加し、同時に全細菌、総乳酸菌、乳酸菌体の各菌数が増加し、大腸菌群数が低下する。
これにより（ａ）整腸効果（ヒトや摂取動物における腸内容物通過時間の短縮と排出物の水分保持性）と共に，水溶性食物繊維がしばしば示す下痢症状の改善効果、及び（ｃ）血清や肝臓中における体内総脂質濃度低下（脂質代謝昂進）効果を確実に得るのである。
つまり前記乳酸菌体とセロビオースとの配合比を１：２．５未満にすると脂質代謝促進効果が得られず、１：１０を超えると下痢症状の改善効果が得られない。
【０００９】
本発明において、乳酸菌体とセロビオースは、単に混合し又は、ペレット化して単品としてよく、またこれらをその他の食品や飼料に添加した配合食・飼料として用いても良い。
【００１０】
前記乳酸菌体とセロビオース以外に配合する他の食品成分や飼料としては、ミネラル混合物、ビタミン混合物、カゼイン、大豆油、ショ糖、ＤＬ−メチオニン等が列記される。
【００１１】
【発明の実施例】
以下に本発明の実施例を詳述する。
本実施例で使用する乳酸菌体は、湿潤状態４℃にて数週間以上安定に生存するもので、−８０℃で冷凍保存した後に調製したものである。この乳酸菌体の調製は、ＭＲＳ培地を用いて３７℃、２４時間、嫌気下に静置培養し、培養後、遠心分離（１０，０００Ｇ、２０ｍｉｎ、４℃）によって集菌し、これを未乾燥のままセロビオースとその他の食品や飼料と配合したもので各飼料組成を表１のＣＥＬ＋乳酸菌に示す。
表１には、５−１０％のセロビオースのみを添加した飼料ＣＥＬと０．５−２．０％の乳酸菌菌体を単独添加した比較例を併記した。
【００１２】
【表１】

【００１３】
実験１：該乳酸菌菌体を単独投与による影響
実験動物として４週齢のウィスター系雄ラットを用いた。ラットは，個別のステンレス製ケージに入れて，室温２３±１℃，１２時間明暗サイクル（明期８：００〜２０：００）下で飼育した。７日間予備飼育（３日間市販固形飼料（ＭＦ：オリエンタル酵母工業（株））、４日間精製飼料）し馴化した後、１群６匹とし、２群に分けた。食物繊維を含まない２０％カゼイン食を基本食（Ｃｏｎｔｒｏｌ）とし、Ｃｏｎｔｒｏｌに乳酸菌菌体を乾燥重量として０．５−２．０％（飼料１ｋｇあたり０．５−２×１０^１３個）添加した実験食の２種類の飼料をそれぞれ水道水とともに自由摂取させ、１４日間飼育した。なお，乳酸菌菌体は未乾燥のまま添加し、乳酸菌体の乾燥重量と同量のショ糖と置換した。
【００１４】
実験２：該乳酸菌菌体およびセロビオースの同時投与による影響
実験動物として３週齢のＷｉｓｔａｒ系雄ラットを用いた。
ラットは、個別のステンレス製ケージに入れて、室温２３±１℃、１２時間明暗サイクル（明期８：００〜２０：００）下で飼育した。３日間市販固形飼料で馴化した後、１群３匹とし、２群に分けた。上記基本食に５−１０％のセロビオースのみを添加した飼料（ＣＥＬ）と、セロビオースと乳酸菌菌体を同時に添加した飼料（ＣＥＬ＋乳酸菌）をそれぞれ水道水とともに自由摂取させ、１４日間飼育した。
実験３：乳酸菌菌体およびセロビオースの同時投与による影響
実験動物として３週齢のＷｉｓｔａｒ系雄ラットを用いた。
ラットは、個別のステンレス製ケージに入れて、室温２３±１℃、１２時間明暗サイクル（明期８：００〜２０：００）下で飼育した。３日間市販固形飼料で馴化した後、１群３匹とし、２群に分けた。上記基本食に５−１０％のセロビオースのみを添加した飼料（ＣＥＬ）と、１０％のセロビオースと乾燥重量として０．５〜２．０％の乳酸菌体を同時に添加した飼料（ＣＥＬ＋乳酸菌）（即ち乳酸菌体対セロビオースの配合比率が１：２．６〜８）をそれぞれ水道水とともに自由摂取させ、１３日間飼育した。飼育期間終了後、エーテル麻酔下で心臓より採血し、肝臓、盲腸を摘出し、重量を測定した。また，盲腸内容物の短鎖脂肪酸量をＨＰＬＣにて、各種細菌菌濃度を１６ＳｒＤＮＡプローブを用いた蛍光ｉｎｓｉｔｕハイブリダイゼーション法にて分析した結果を表６に記載してある。
結果
実験１および実験２において，１４日間の飼育を行ったときの体重増加量，飼料摂取量，飼料効率，体重１００ｇあたりの臓器重量を表２，表３に示す。
【００１５】
【表２】

【００１６】
【表３】

【００１７】
いずれにおいても各二群間に有意差はなく、乳酸菌菌体の添加による体重増加および臓器重量比に対する影響は認められなかった。
【００１８】
乳酸菌菌体単独投与の血清脂質に及ぼす影響についての結果を表４に示す。
Ｃｏｎｔｒｏｌに対し，乳酸菌菌体単独投与の群の各数値には、ほとんど差がなく乳酸菌菌体のみによる影響は観察されなかった。
乳酸菌菌体とセロビオースの同時投与による血清および肝臓脂質への影響について表５に示す。
血清成分に関しては、乳酸菌体とセロビオースを配合のＣＥＬ＋乳酸菌投与の群の値はＣＥＬ投与の群の値と比較して、総脂質、中性脂肪、β−リポタンパクにおいて有意な低値を示した。総コレステロール、コレステロールおよびリン脂質は有意差はないものの低い値を示す傾向がみられた。また肝臓脂質についてもＣＥＬと投与の群に対して、乳酸菌体とセロビオースを配合のＣＥＬ＋乳酸菌投与の群の値は減少する傾向を示し、乳酸菌菌体の単独投与では認められなかった優れた効果現象が観察された。
【００１９】
【表４】

【００２０】
【表５】

【００２１】
以上のことから、実験３における盲腸内容物の短鎖脂肪酸量をＨＰＬＣ（：有機酸分析システム，島津製作所製）にて、各種細菌菌濃度を１６ＳｒＤＮＡプローブを用いた蛍光ｉｎｓｉｔｕハイブリダイゼーション法にて分析した結果を表６に示す。
その結果、乳酸菌体（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｈａｍｎｏｓｕｓ）とセロビオースを配合のＣＥＬ＋乳酸菌の投与の群ではｐＨの低下と共に酢酸及びプロピオン酸量が有意に増加しており、同時に全細菌、総乳酸菌、該乳酸菌の各菌数が増加し、大腸菌群数が若干低下していた。
【００２２】
【表６】

【００２３】
【発明の効果】
以上の説明で明らかなように、本発明の乳酸菌（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｈａｍｎｏｓｕｓ）菌体とセロビオースを配合せしめた整腸及び脂質代謝促進用肥料食・飼料添加物は、（ａ）整腸効果（ヒトや摂取動物における腸内容物通過時間の短縮と排出物の水分保持性）と共に，水溶性食物繊維がしばしば示す下痢症状の改善効果，及び（ｂ）血清や肝臓中における体内総脂質濃度低下（脂質代謝昂進）効果を有する。
【００２４】
人は様々なストレスから下痢や便秘などを起こす。また、昨今の西欧型食事即ち脂質摂取量過多による肥満とそれに由来する成人病発症が問題となっている。また、ペットなどにおいてはストレスを与える生育環境、不衛生畜舎、投与飼料のインバランスその他を原因とする下痢や便秘などがしばしば引き起こされる。これに対して抗生剤等の予防的投与が行われているがそのような無節操抗生剤の使用は畜産製品への残留の問題と土壌環境中などへの過剰分の放出による抗生物質耐性菌の出現という問題を抱えている。
このような場面においてゆるやかで，しかも常食，過剰摂取に関して問題点が極めて少ないと考えられる本発明品は，特定健康保健食品あるいは機能性ペットフードとしての使用が今後大いに期待される。[0001]
BACKGROUND OF THE INVENTION
TECHNICAL FIELD The present invention relates to a fertilizer food / feed additive for promoting lipid metabolism, which exhibits a significant effect of lowering the lipid concentration as compared with the case of each of the intestinal effects by containing lactic acid bacteria and water-soluble dietary fiber simultaneously. It is.
[0002]
[Prior art]
The report that has been interested in the functions of lactic acid bacteria and fermented milk has been said to be the longevity theory proposed by Metnikov in the early 20th century. This is because people who regularly eat fermented milk have a long life expectancy, because lactic acid bacteria colonize the intestines and prevent spoilage by harmful bacteria.
[0003]
In recent years, much attention has been paid from the viewpoint of probiotics / biogenics, and many studies on lactic acid bacteria as functional food ingredients have been conducted.
[0004]
So far, reports on functionality have been known such as intestinal regulating action, blood cholesterol lowering action, antitumor action, immunostimulatory action, blood pressure lowering action and the like. However, these are all examples taken with lactic acid bacteria alone.
On the other hand, it is also widely known that certain types of water-soluble dietary fiber exhibit effects such as intestinal regulation, inhibition of intestinal rot, and colon cancer.
[0005]
[Problems to be solved by the invention]
However, the water-soluble dietary fiber has a problem of causing diarrhea symptoms by excessive administration.
In addition, there have been no reports on oligosaccharides or microbial preparations that can promote metabolism promotion for all lipids in serum and liver.
[0006]
[Means for Solving the Problems]
The present invention is an excellent solution that solves the above-mentioned problems, and is characterized in that lactic acid bacteria (Lactobacillus rhamnosus) and cellobiose are contained in a mixing ratio of 1: 2.5-10. It is in fertilizer food and feed additive for promoting intestinal regulation and lipid metabolism.
[0007]
DETAILED DESCRIPTION OF THE INVENTION
That is, the present inventors have examined the effect on the lipid metabolism of a lactic acid bacterium, Lactobacillus rhamnosus (hereinafter simply referred to as “lactic acid bacterium”), which has been isolated and identified, with the expectation of its functional expression. Administration did not affect serum lipids, but when lactic acid bacteria were administered at the same time with cellobiose, serum total lipid, neutral fat, β-lipoprotein and cholesterol levels decreased, as well as liver lipids. The present invention has been completed by obtaining new knowledge of the synergistic effect.
[0008]
In the present invention, the mixing ratio of the lactic acid bacteria and cellobiose is 1: 2.5 to 8 so that the amount of acetic acid and propionic acid is significantly increased with the decrease in pH, and at the same time, all bacteria, total lactic acid bacteria, lactic acid bacteria The number of each bacteria increases and the number of coliforms decreases.
As a result, (a) an intestinal regulation effect (shortening of the transit time of intestinal contents and water retention of excreta in humans and ingestion animals), improvement of diarrhea symptoms often exhibited by water-soluble dietary fiber, and (c) serum and Thus, the effect of lowering the total body lipid concentration (facilitating lipid metabolism) in the liver is reliably obtained.
That is, if the blending ratio of the lactic acid bacteria and cellobiose is less than 1: 2.5, the lipid metabolism promoting effect cannot be obtained, and if it exceeds 1:10, the diarrhea symptom improving effect cannot be obtained.
[0009]
In the present invention, the lactic acid bacteria and cellobiose may be simply mixed or pelletized to form a single product, or they may be used as a mixed food or feed added to other food or feed.
[0010]
As other food ingredients and feeds to be blended in addition to the lactic acid bacteria and cellobiose, mineral mixtures, vitamin mixtures, casein, soybean oil, sucrose, DL-methionine and the like are listed.
[0011]
DETAILED DESCRIPTION OF THE INVENTION
Examples of the present invention are described in detail below.
The lactic acid bacterium used in this example stably survives for several weeks or more in a wet state at 4 ° C., and is prepared after being stored frozen at −80 ° C. The lactic acid bacterial cells were prepared by static culture under anaerobic conditions using MRS medium at 37 ° C. for 24 hours, and then collected by centrifugation (10,000 G, 20 min, 4 ° C.). The composition of each feed is shown as CEL + lactic acid bacteria in Table 1 by mixing cellobiose with other foods and feeds in the dry state.
Table 1 also shows a feed CEL to which only 5-10% cellobiose was added and a comparative example in which 0.5-2.0% lactic acid bacteria were added alone.
[0012]
[Table 1]

[0013]
Experiment 1: Effect of single administration of the lactic acid bacteria cells Four-week-old Wistar male rats were used as experimental animals. Rats were housed in individual stainless steel cages at room temperature 23 ± 1 ° C. under a 12-hour light / dark cycle (light period 8:00 to 20:00). After 7 days of preliminary breeding (3-day commercial solid feed (MF: Oriental Yeast Co., Ltd.), 4-day purified feed) and acclimatization, each group was divided into 2 groups. A 20% casein diet containing no dietary fiber was used as a basic diet (Control), and 0.5 to 2.0% (0.5-2 × 10 ¹³ cells per kg of feed) of lactic acid bacteria were added to Control as a dry weight. Two types of feed for the experimental food were freely ingested together with tap water, and were reared for 14 days. The lactic acid bacterial cells were added undried and replaced with the same amount of sucrose as the dry weight of the lactic acid bacterial cells.
[0014]
Experiment 2: Effect of simultaneous administration of the lactic acid bacteria and cellobiose A 3-week-old Wistar male rat was used as an experimental animal.
Rats were housed in individual stainless steel cages at room temperature 23 ± 1 ° C. under a 12-hour light / dark cycle (light period 8:00 to 20:00). After acclimatization with commercially available solid feed for 3 days, 3 animals per group were divided into 2 groups. A feed (CEL) to which only 5-10% cellobiose was added to the basic diet and a feed (CEL + lactic acid bacteria) to which cellobiose and lactic acid bacteria were added at the same time were freely ingested together with tap water and reared for 14 days.
Experiment 3: Effect of simultaneous administration of lactic acid bacteria and cellobiose A 3-week-old Wistar male rat was used as an experimental animal.
Rats were housed in individual stainless steel cages at room temperature 23 ± 1 ° C. under a 12-hour light / dark cycle (light period 8:00 to 20:00). After acclimatization with commercially available solid feed for 3 days, 3 animals per group were divided into 2 groups. Feed (CEL) in which only 5-10% cellobiose is added to the basic diet, and feed (CEL + lactic acid bacteria) in which 10% cellobiose and 0.5 to 2.0% lactic acid bacteria are simultaneously added as dry weight (ie, CEL + lactic acid bacteria) The mixing ratio of lactic acid bacteria to cellobiose was 1: 2.6 to 8), which was freely ingested with tap water, and bred for 13 days. After the breeding period, blood was collected from the heart under ether anesthesia, and the liver and cecum were removed and weighed. Table 6 shows the results of analyzing the amount of short-chain fatty acids in the cecum contents by HPLC and the concentration of various bacteria by fluorescent in situ hybridization using a 16S rDNA probe.
Results Tables 2 and 3 show the body weight gain, feed intake, feed efficiency, and organ weight per 100 g body weight in 14 days of breeding in Experiment 1 and Experiment 2.
[0015]
[Table 2]

[0016]
[Table 3]

[0017]
In either case, there was no significant difference between the two groups, and no effect on body weight increase and organ weight ratio by addition of lactic acid bacteria was observed.
[0018]
Table 4 shows the results of the effect of lactic acid bacteria alone on serum lipids.
There was almost no difference in each numerical value of the group in which lactic acid bacteria were administered alone with respect to Control, and no influence due to lactic acid bacteria alone was observed.
Table 5 shows the influence on serum and liver lipids caused by simultaneous administration of lactic acid bacteria and cellobiose.
Regarding serum components, the values of the CEL + lactic acid bacteria administration group containing lactic acid bacteria and cellobiose showed significantly lower values in total lipid, neutral fat, and β-lipoprotein than the values of the CEL administration group. . Total cholesterol, cholesterol and phospholipids tended to show low values although there was no significant difference. In addition, for liver lipids, the value of the CEL + lactic acid bacteria administration group containing lactic acid bacteria and cellobiose tended to decrease compared to the CEL and administration groups, and an excellent effect phenomenon that was not observed with the administration of lactic acid bacteria alone Was observed.
[0019]
[Table 4]

[0020]
[Table 5]

[0021]
Based on the above, the amount of short-chain fatty acids in the cecum contents in Experiment 3 was determined by HPLC (: organic acid analysis system, manufactured by Shimadzu Corporation), and the concentration of various bacteria was determined by fluorescence in situ hybridization using a 16S rDNA probe. Table 6 shows the results of analysis.
As a result, the amount of acetic acid and propionic acid significantly increased with the decrease in pH in the group of CEL + lactic acid bacteria containing lactic acid bacteria (Lactobacillus rhamnosus) and cellobiose, and at the same time, all bacteria, total lactic acid bacteria, The number increased and the number of coliforms slightly decreased.
[0022]
[Table 6]

[0023]
【The invention's effect】
As is apparent from the above description, the fertilizer food / feed additive for promoting intestinal and lipid metabolism, which is a combination of lactic acid bacteria of the present invention and cellobiose, has the following effects: (B) Shortening of intestinal content transit time and water retention of excreta in animals, as well as improvement of diarrhea symptoms often exhibited by water-soluble dietary fiber, and (b) Decrease in total body lipid level in serum and liver (increased lipid metabolism) ) Has an effect.
[0024]
People cause diarrhea and constipation from various stresses. Further, obesity due to recent Western-style meals, that is, excessive intake of lipids, and the onset of adult diseases resulting therefrom have been a problem. In pets and the like, diarrhea and constipation are often caused by a stressful growth environment, an unsanitary barn, an imbalance in the administered feed, and the like. In contrast to this, antibiotics and other preventive administrations have been carried out. However, the use of such artillery antibiotics is a problem of residues in livestock products and the release of antibiotic-resistant bacteria due to excessive release into the soil environment. I have a problem of appearance.
In this situation, the product of the present invention, which is considered to be gradual and has few problems with regular diet and excessive intake, is expected to be used as a specific health health food or functional pet food.

Claims

乳酸菌（Ｌａｃｔｏｂａｃｉｌｌｕｓｒｈａｍｎｏｓｕｓ）菌体とセロビオースを配合比で、１：２．５〜８含有せしめたことを特徴とする整腸及び脂質代謝促進用肥料食・飼料添加物。A fertilizer food and feed additive for promoting intestinal regulation and lipid metabolism, comprising lactic acid bacteria (Lactobacillus rhamnosus) and cellobiose in a mixing ratio of 1: 2.5-8.