JP2004321068A5 - - Google Patents

Description

【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0.5-2×10¹³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：該乳酸菌の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
オースと乳酸菌の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：乳酸菌の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16S rDNAプローブを用いた蛍光in situ 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、乳酸菌の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】
乳酸菌の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，乳酸菌の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。
乳酸菌の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Ｃ
ＥＬ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、乳酸菌の菌
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16S rDNAプローブを用いた蛍光in situ 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(Lactobacillus rhamnosus)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 (Lactobacillus rhamnosus)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。




[Document Name] Statement
[Title of the Invention]
Food and feed additives for promoting lipid metabolism [Claims]
(1)
A food / feed additive for promoting intestinal regulation and lipid metabolism, characterized by containing lactic acid bacteria (Lactobacillus rhamnosus) cells and cellobiose in a mixing ratio of 1: 2.5-8.
DETAILED DESCRIPTION OF THE INVENTION
[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a food / feed additive for promoting lipid metabolism, which simultaneously contains a lactic acid bacterium and water-soluble dietary fiber, thereby exhibiting an intestinal effect and a markedly lower lipid concentration effect as compared to the case of using each alone. It is.
[0002]
[Prior art]
A report that has triggered interest in the functions of lactic acid bacteria and fermented milk has long been called the theory of longevity proposed by Mechnikov in the early 20th century. This was thought to be due to the long lifespan of people who regularly eat fermented milk, because lactic acid bacteria settle in the intestine and control spoilage by harmful bacteria.
[0003]
In recent years, much attention has been paid from the viewpoint of probiotics and biogenics, and many studies on lactic acid bacteria as functional food ingredients have been made.
[0004]
So far, reports on functionality include intestinal action, blood cholesterol lowering action, antitumor action, immunostimulatory action, blood pressure lowering action, and the like. However, these are all examples of lactic acid bacteria taken alone.
On the other hand, it is widely known that certain water-soluble dietary fibers exhibit effects such as intestinal regulation, intestinal putrefaction suppression, and colorectal cancer suppression.
[0005]
[Problems to be solved by the invention]
However, the water-soluble dietary fiber has a problem of causing diarrhea symptoms due to excessive administration.
In addition, there have been no reports on oligosaccharides or microbial preparations capable of promoting the metabolism of all lipids in serum and liver.
[0006]
[Means for Solving the Problems]
The present invention is excellent in solving the above-mentioned problems, and is characterized by lactic acid bacteria (
Lactobacillus rhamnosus) is a food / feed additive for promoting intestinal regulation and lipid metabolism, characterized by containing cells and cellobiose in a mixing ratio of 1: 2.5 to 10.
[0007]
BEST MODE FOR CARRYING OUT THE INVENTION
That is, the present inventors examined the effects of isolated and identified lactic acid bacteria Lactobacillus rhamnosus (hereinafter simply referred to as lactic acid bacteria) on lipid metabolism by administering them to rats in hope of expressing their functionality. There was no effect on serum lipids after administration, but administration of lactic acid bacteria simultaneously with cellobiose decreased serum total lipids, neutral fats, β-lipoprotein and cholesterol levels, and also reduced liver lipids. Thus, the present invention has been completed by obtaining new knowledge of the synergistic effect.
[0008]
In the present invention, the compounding ratio of the lactic acid bacteria and cellobiose is 1: 2.5 to 8 to significantly increase the amount of acetic acid and propionic acid with a decrease in pH, and at the same time, all bacteria, total lactic acid bacteria,
The number of each lactic acid bacteria increases and the number of coliforms decreases.
This results in (a) an intestinal control effect (shortening of intestinal contents transit time in humans and ingested animals and water retention of excretion), (b) an improvement effect of diarrhea symptoms often exhibited by water-soluble dietary fiber, and (c) ) The effect of lowering the total lipid concentration in the body (enhancing lipid metabolism) in serum and liver is surely obtained.
In other words, when the mixing ratio of the lactic acid bacteria and cellobiose is less than 1: 2.5
The effect of promoting lipid metabolism cannot be obtained, and if it exceeds 1:10, the effect of improving diarrhea symptoms 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 may be used as a compound food or feed obtained by adding these to other foods or feeds.
[0010]
Other food ingredients and feeds to be added in addition to the lactic acid bacteria and cellobiose include mineral mixtures, vitamin mixtures, casein, soybean oil, sucrose, DL-methionine, and the like.
[0011]
DESCRIPTION OF THE PREFERRED EMBODIMENTS
Hereinafter, examples of the present invention will be described in detail.
The lactic acid bacteria used in this example are those that survive several weeks or more at 4 ° C. in a humid state.
Prepared after freezing at 0 ° C. The lactic acid bacteria were prepared by statically culturing at 37 ° C. for 24 hours in an MRS medium under anaerobic conditions.After culturing, the cells were collected by centrifugation (10,000 G, 20 min, 4 ° C.). The composition of each feed, which was obtained by mixing cellobiose with other foods and feeds, is shown in Table 1 as CEL + lactic acid bacteria.
Table 1 also shows a feed CEL containing only 5-10% cellobiose and a comparative example containing only 0.5-2.0% lactic acid bacteria cells.
[0012]
[Table 1]

[0013]
Experiment 1: Influence of single administration of the lactic acid bacteria cells Wistar male rats of 4 weeks 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: 00-20: 00). After preliminarily rearing for 7 days (commercially available solid feed (MF: Oriental Yeast Co., Ltd.) for 3 days and purified feed for 4 days), the animals were divided into 2 groups, each group consisting of 6 animals. 20% casein diet without dietary fiber was used as the basic diet (Control), and two types of experimental diets were prepared by adding 0.5-2.0% (0.5-2 × 10 ¹³ per kg of feed) lactic acid bacteria cells to the control. Each feed was freely fed with tap water and bred for 14 days. The lactic acid bacterial cells were added in an undried state and replaced with sucrose in the same amount as the dry weight of the lactic acid bacterial cells.
[0014]
Experiment 2: Effect of simultaneous administration of the cells of the lactic acid bacterium and cellobiose Male Wistar rats, 3 weeks old, 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 acclimation with a commercial solid feed for 3 days, the animals were divided into 2 groups, each group consisting of 3 animals. A feed (CEL) in which only 5-10% cellobiose was added to the above-mentioned basic diet and a feed (CEL + lactic acid bacteria) in which cells of cellobiose and lactic acid bacteria were simultaneously added were allowed to freely ingest with tap water, and bred for 14 days.
Experiment 3: Effect of simultaneous administration of cells of lactic acid bacteria and cellobiose Male 3-year-old Wistar 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 acclimation with a commercial solid feed for 3 days, the animals were divided into 2 groups, each group consisting of 3 animals. A feed (CEL) in which only 5-10% cellobiose is added to the above basic diet, and a feed (CEL + lactic acid bacteria) in which 10% cellobiose and 0.5-2.0% lactic acid bacteria are added at the same time as a dry weight (ie, lactic acid bacteria versus cellobiose) Were mixed with tap water and freely fed, and bred for 13 days. After the end of the breeding period, blood was collected from the heart under ether anesthesia, the liver and cecum were removed, and the weight was measured. Table 6 shows the results of analysis of the content of short-chain fatty acids in the contents of the cecum by HPLC and the concentration of various bacteria by fluorescence in situ hybridization using a 16S rDNA probe.
Results In Experiments 1 and 2, Tables 2 and 3 show the weight gain, feed intake, feed efficiency, and organ weight per 100 g of body weight when bred for 14 days.
[0015]
[Table 2]

[0016]
[Table 3]

[0017]
In each case, there was no significant difference between the two groups, and the addition of lactic acid bacteria did not affect the weight gain or organ weight ratio.
[0018]
Table 4 shows the results of the effects of the administration of the lactic acid bacteria alone on the serum lipid.
There was almost no difference between Control and each value in the group administered with the lactic acid bacteria alone, and no effect of the lactic acid bacteria alone was observed.
Effect of simultaneous administration of lactic acid bacteria cells and cellobiose on serum and liver lipids Table 5
Shown in
Regarding serum components, the value of the group administered with CEL + lactic acid bacteria containing lactic acid bacteria and cellobiose was C
Compared with the value of the group to which EL was administered, significantly lower values were shown in total lipids, neutral fats, and β-lipoprotein. Total cholesterol, cholesterol and phospholipids tended to show low values, though not significantly different. As for the liver lipid, the value of the CEL + lactic acid bacteria administration group containing lactic acid bacteria and cellobiose showed a tendency to decrease with respect to the CEL and administration groups, and an excellent effect that was not observed when the lactic acid bacteria was administered alone. A phenomenon was observed.
[0019]
[Table 4]

[0020]
[Table 5]

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

[0023]
【The invention's effect】
As is clear from the above description, the food and feed additives for promoting intestinal and lipid metabolism in which the lactic acid bacteria (Lactobacillus rhamnosus) cells of the present invention and cellobiose are blended have the following effects. Water-soluble dietary fiber often improves diarrheal symptoms, and (b) lowers total lipid levels in the serum and liver (enhanced lipid metabolism) Has an effect.
[0024]
People suffer from various stresses such as diarrhea and constipation. In addition, obesity due to the recent Western-style diet, that is, excessive fat intake, and the onset of adult diseases caused by it have become a problem.
In pets and the like, diarrhea and constipation due to stressed growth environments, unsanitary stables, imbalance of the feed to be administered, and the like are often caused.
On the other hand, prophylactic administration of antibiotics and the like has been performed, but the use of such uncontrolled antibiotics has led to the problem of residue on livestock products and the release of antibiotic-resistant bacteria due to excessive release into the soil environment. Have the problem of appearance.
In such a situation, the product of the present invention, which is considered to be slow and has very few problems with normal eating and excessive intake, is expected to be used as a specified health and health food or a functional pet food in the future.