Feed composition and method for producing same

Abstract

To provide novel means of improving animal gut flora and enhancing animal growth.SOLUTION: Provided is a feed composition comprising loliose or loliose disaccharide.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a feed composition, a method for producing the feed composition, and a composition for microbial growth.

Background Art

[0002] Hitherto, in the livestock industry, administration of antibacterial substances such as antibacterial feed additives has been routinely performed for improving livestock productivity and preventing infectious diseases. However, it has become clear that the misuse of such antibacterial substances is a factor leading to the frequent emergence of drug-resistant bacteria. In 2017, the World Health Organization advised against the use of antibiotics in healthy animals for growth promotion and disease prevention. As a result, in the production field, a decrease in productivity due to an increase in morbidity has been regarded as a problem, and improvement of livestock productivity has become an important issue. In order to solve such problems, development of products that contribute to improving livestock productivity as an alternative to antibacterial substances is expected, and attention has been focused on the development of feed additives expected to have an effect of improving the intestinal flora.

[0003] It is well known that adding prebiotics such as oligosaccharides to human food and livestock feed improves the intestinal flora. For example, Patent Document 1 describes an animal feed composition composed of an oligosaccharide composition, and it is described that this animal feed composition promotes the growth of animals. Further, Patent Document 2 describes a livestock feed containing a saccharide having fructooligosaccharide as a main component, and it is described that this livestock feed improves diarrhea frequently seen at the time of weaning of livestock and improves weight gain efficiency.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0005] Since the addition of prebiotics and the like to feed directly leads to an increase in the production cost of livestock products, the development of new prebiotic materials with excellent cost performance is required. The present invention has been made under such a background, and an object thereof is to provide a novel means for improving the intestinal flora of animals and promoting the growth of animals.

Means for Solving the Problems

[0006] The present inventor focused on oligosaccharides called loliose, which are characteristically contained in plants of the genus Lolium and the genus Festuca used as forage, and found that a seed pulverized product of ryegrass, which is a kind of plant of the genus Lolium, exhibits a growth promoting effect on chicks of chickens. Based on this finding, the present invention has been completed.

[0007] That is, the present invention provides the following [1] to [7]. [1] A feed composition characterized by containing loliose or loliobiose.

[0008] [2] The feed composition according to [1], which is for livestock or poultry.

[0009] [3] The feed composition according to [1], which is for chickens.

[0010] [4] A method for producing a feed composition containing loliose or loliobiose, the method for producing a feed composition being characterized by including the following steps (1) to (3): (1) A step of removing husks from seeds of one or more plants selected from the group consisting of plants of the genus Lolium and plants of the genus Festuca; (2) A step of pulverizing the seeds obtained in step (1); (3) A step of recovering a seed pulverized product passing through a sieve with an opening size of 0.5 to 2.0 mm from the seed pulverized product obtained in step (2).

[0011] The method for producing the feed composition according to [4], further comprising a step of recovering the seed pulverized product that does not pass through a sieve with an opening size of 100 to 200 μm from the seed pulverized product obtained in the step [5](4)(3).

[0012] The composition for promoting microbial growth, characterized by containing loliolose or loliolose disaccharide.

[0013] The composition for promoting microbial growth according to [6], characterized in that the microorganism is Bifidobacterium.

Advantages of the Invention

[0014] The present invention provides a novel feed composition, a novel method for producing a feed composition, and a novel composition for promoting microbial growth.

Brief Description of the Drawings

[0015]

Figure 1

Embodiments for Carrying Out the Invention

[0016] Hereinafter, the present invention will be described in detail. (A) Feed composition The feed composition of the present invention is characterized by containing loliolose or loliolose disaccharide.

[0017] In the present invention, the "feed composition" usually means a composition for addition to various feeds, but in some cases, the feed composition itself may be a feed.

[0018] Raffinose is a trisaccharide in which a galactose residue is α1,3-bonded to the glucose residue of sucrose and is an isomer of raffinose. Raffinose is known to be present in perennial ryegrass (Lolium perenne L.) and meadow fescue (Festuca pratensis Huds.), which are forage grasses, but its dynamics, growth and decline, and physiological roles in plants are not well understood.

[0019] The raffinose used may be purified or may contain impurities. As raffinose containing impurities, for example, a pulverized product of seeds of a plant containing raffinose can be used. The pulverized product of seeds of a plant containing raffinose can be prepared, for example, according to the method for producing a feed composition described below. Also, purified raffinose can be prepared by extracting and purifying the aforementioned seed pulverized product.

[0020] Raffinobiose is a disaccharide (α-D-galactosyl-(1,3)-D-glucose) in which galactose and glucose, which are produced by decomposing the fructofuranosyl bond of raffinose and removing the fructose residue, are α1,3-bonded and is an isomer of melibiose. There are few research examples on raffinobiose and no common name has been given.

[0021] The raffinobiose used may be purified or may contain impurities, similar to raffinose. Raffinobiose can be prepared by subjecting raffinose to acid treatment.

[0022] The feed composition of the present invention may contain components other than raffinose and raffinobiose.

[0023] When feeding an animal with a feed to which the feed composition of the present invention is added, the content of raffinose or raffinobiose in the feed is not particularly limited, but can be, for example, 0.01 to 10% by weight, preferably 0.1 to 1% by weight.

[0024] The type of animal to which the feed composition of the present invention is administered is not particularly limited, but poultry (for example, chickens, quails, turkeys, geese, ducks, etc.) and livestock (for example, cows, pigs, sheep, etc.) are preferred, and chickens are particularly preferred.

[0025] As shown in the examples described below, loliolose shows a growth-promoting effect on chickens. The reason for this is not necessarily clear, but since loliolose has a Bifidobacterium growth-promoting effect (Figure 1), it is presumed to be due to the improvement of the intestinal flora of chickens. Although it has not been confirmed at present that loliobiose shows a growth-promoting effect on chickens, loliobiose also has a Bifidobacterium growth-promoting effect like loliolose (Figures 1A - D), so it is presumed that loliobiose also shows a growth-promoting effect on chickens. In addition, the growth-promoting effects of loliolose and loliobiose have been confirmed not only for Bifidobacterium isolated from the ceca of chickens (Figure 1A), but also for Bifidobacterium isolated from pig feces (Figure 1D) and bovine rumen (Figure 1B). Therefore, it is presumed that loliolose and loliobiose show a growth-promoting effect on animals other than chickens, such as cows and pigs.

[0026] (B) Method for producing the feed composition The method for producing a feed composition containing loliolose or loliobiose of the present invention is characterized by including the following steps (1) to (3), and more preferably, further characterized by including the following step (4).

[0027] In step (1), the husk is removed from the seeds of one or more plants selected from the group consisting of plants of the genus Lolium and plants of the genus Festuca. Examples of plants of the genus Lolium include Italian ryegrass (Lolium multiflorum Lam.), hybrid ryegrass (Lolium x hybridum Hausskn.), perennial ryegrass (Lolium perenne L.), darnel (Lolium temulentum L.), etc. Examples of plants of the genus Festuca include meadow fescue (Festuca pratensis Huds.), tall fescue (Festuca arundinacea Schreb.), sheep fescue (Festuca ovina L.), etc. The seeds to be used may be only one of these plants, or seeds mixed with two or more of them may be used. The removal of the husk can be carried out according to the method commonly used in the removal of the husk of seeds of Gramineae plants.

[0028] In step (2), the seeds obtained in step (1) are ground. The grinding can be carried out according to the method commonly used in the grinding of seeds of Gramineae plants.

[0029] In step (3), the ground seed product that passes through a sieve (first sieve) with a mesh size of 0.5 to 2.0 mm is recovered from the ground seed product obtained in step (2). As the first sieve, a sieve with a mesh size of 0.5 to 2.0 mm may be used, but it is preferable to use a sieve with a mesh size of 0.8 to 1.5 mm, and more preferably to use a sieve with a mesh size of 1.0 mm.

[0030] In step (4), seed pulverized matter that does not pass through a sieve (second sieve) with an opening size of 100 to 200 μm is recovered from the seed pulverized matter obtained in step (3). As the second sieve, a sieve with an opening size of 100 to 200 μm may be used, but it is preferable to use a sieve with an opening size of 150 to 200 μm, and more preferably to use a sieve with an opening size of 180 μm. By thus recovering the seed pulverized matter that has passed through the first sieve and does not add the second sieve, it is possible to obtain a seed pulverized matter that contains a large amount of loliose and has few impurities.

[0031] (C) Composition for Microorganism Growth The composition for microorganism growth of the present invention is characterized by containing loliose or loliose disaccharide. Here, the "composition for microorganism growth" refers to a composition used for growing specific microorganisms in the body of an administration target animal (human or non-human animal). The type of microorganism is not particularly limited, but microorganisms that bring beneficial effects to the host present in the digestive tract such as the intestine and rumen are preferable, and examples thereof include Bifidobacterium and lactic acid bacteria.

[0032] The loliose and loliose disaccharide to be used may be purified or may contain impurities, similar to the feed composition.

[0033] The composition for microorganism growth of the present invention may contain components other than loliose and loliose disaccharide, such as excipients, emulsifiers, stabilizers, pH adjusters, flavors, and the like. Further, when the composition for microorganism growth of the present invention is used as a nutritional composition, a food composition, or a pharmaceutical composition, it may contain components usually contained in each composition.

[0034] The intake or dosage of the composition for microorganism growth of the present invention is not particularly limited as long as it can grow the target microorganism. For example, in the case of growing Bifidobacterium, the intake or dosage per human adult per day can be 0.01 to 100 mg of loliose or loliose disaccharide per kg of body weight, and preferably 1 to 10 mg.

Example

[0035] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

[0036] [Example 1] (1) Method (1.1) Preparation of oligosaccharide material Italian ryegrass seeds containing loliosaccharide were prepared using commercially available Wase Aoba (Snow Brand Seed Co., Ltd.). For the palatability test, the seeds themselves were used, and for the first feeding test, ryegrass seeds (powder) ground with a 1 mm mesh screen using an ultracentrifugal mill (ZM-100, RETSCH) were used. Also, as described later in (2.2), since energy deficiency was suggested in the ryegrass seeds (powder), in the second feeding test, scutching by low-speed stirring was performed before grinding, and the fraction that passed through a 180 μm sieve after grinding was removed and used as ryegrass seeds (flour). Thus, a composition with unnecessary components removed while maintaining the loliosaccharide content was used for the feeding test.

[0037] Lacto-N-bios I (LNB) was prepared using a high-purity powder (purity 99%) prepared according to a previously developed method (M. NISHIMOTO and M. KITAOKA: Practical Preparation of Lacto-N-biose I, the Candidate of the Bifidus Factor in Human Milk. Biosci. Biotechnol. Biochem., 71 (8), 2101-2104 (2007)).

[0038] (1.2) Palatability test Ten 31-day-old male Rhode Island Red chicks were used for the preference test. The chicks were housed in five cages, with two chicks per cage, so that their average weights were approximately equal. In each cage, a feeding box was installed so that the chicks could freely consume both a commercially available feed for laying hen chicks (control feed, Adjusted Chicks, Toyohashi Feed Co., Ltd.) and a feed prepared by mixing the control feed and ryegrass seeds in equal amounts (ryegrass feed (granules)). The test period was six days. The position of the feeding box was changed daily so that on odd days, the control feed was on the left and the ryegrass feed (granules) was on the right, and on even days, it was the reverse. The intake of each feed was measured over the six-day period.

[0039] (1.3) First Feeding Test Thirty-five-day-old male White Leghorn chicks were used. Three types of feeds were prepared: a control feed mainly composed of corn and soybean meal (Table 1) designed to meet the nutrient requirements during the growing chick stage of laying hens, a feed prepared by mixing the control feed and ground ryegrass seeds in equal amounts (ryegrass feed (powder)), and a feed with 1% LNB added to the control feed. Five chicks with equal average weights were assigned to each feed and fed for eight days. The metabolic energy (ME) content and ileal amino acid digestibility of the mixed ryegrass seeds were determined by the index method using chromium oxide.

Table 1

[0040] (1.4) Second Feeding Test Sixteen-day-old male Yellow-Spotted Plymouth Rock chicks were used. Three types of feeds were prepared: a control feed mainly composed of corn and soybean meal designed to meet the nutrient requirements during the growing chick stage of laying hens, a ryegrass feed (flour) prepared by mixing 50% of the ryegrass seeds purified as described in (1.1) and adjusted so that the nutrient content was equivalent to that of the control feed (Table 1), and feeds with 0.1% and 1.0% LNB added. Five chicks with equal average weights were assigned to each feed and fed for 16 days.

[0041] (1.5) Bifidobacterium Culture Test The utilization properties of various oligosaccharides of reference strains of Bifidobacterium species (obtained from the Microbial Materials Development Laboratory, RIKEN BioResource Center) were investigated. After pre-culturing under anaerobic conditions for 24 hours using a liquid medium prepared by adding Phytone peptone (5 g / L) and L-cysteine HCl (0.5 g / L) to Man-Rogosa-Sharpe medium (55 g / L), the sugar-source free basal medium [tryptone (5.0 g / L), yeast extract (5.0 g / L), peptone (10 g / L), K 2 HPO 4 (2.0 g / L), diammonium citrate (2.0 g / L), sodium acetate (5.0 g / L), manganese sulfate monohydrate (0.25 g / L), manganese sulfate monohydrate (0.58 g / L), Tween 80 (1 ml / L) and L-cysteine HCl (0.5 g / L)] was added with various oligosaccharides at a final concentration of 1% (w / v), and anaerobic culture was performed for 72 hours. The utilization properties of oligosaccharides of each strain were determined by measuring the pH and turbidity (OD600) of the culture supernatant after the culture was completed.

[0042] As the reference strains of Bifidobacterium species, six strains were used: B. pullorum subsp. gallinarum JCM 6291T (isolation source: chicken cecum), B. ruminantium JCM 8222T (isolation source: bovine rumen), B. catenulatum subsp. catenulatum JCM 1194T (isolation source: human feces), B. longum subsp. suillum JCM 19995T (isolation source: piglet feces), B. thermacidophilum subsp. thermacidophilum JCM 11165T (isolation source: sewage), and B. boum JCM 1211T (isolation source: bovine rumen). In addition, the oligosaccharides used were arabinose (Ara), glucose (Glu), fructooligosaccharide (FOS), raffinose (Raf), lactose (Lac), galacto-N-biose (GNB), LNB, fesculose disaccharide (Fes), loliolose (Lol), and trehalose (Tre).

[0043] (1.6) Preparation of Purified Lolioside Standard (1) Commercially available perennial ryegrass seeds (600 g) were ground with a mill, and 2.4 L of 80% (v / v) ethanol was added to dissolve the oligosaccharide fraction. After the suspension was fractionated into a filtrate and a precipitate by filtration, 2.4 L of 80% (v / v) ethanol was added again to the precipitate for re-extraction. The obtained filtrate (4.6 L) was concentrated to 1.4 L with an evaporator, and 2 L of water was added and filtered to remove the insoluble fraction. The obtained filtrate was subjected to microfiltration (PMP003 membrane, Asahi Kasei) and ultrafiltration (SLP0053 membrane, Asahi Kasei) to remove impurities with a molecular weight of 10,000 or more. In addition, a DEAE-cellulose carrier was added to the obtained filtrate (2.5 L), stirred, and filtered to adsorb and remove negatively charged impurities on the carrier. The obtained filtrate (3.3 L) was concentrated to 80 mL with an evaporator and subjected to gel filtration (carrier TOYOPEARL HW40S, Tosoh, column diameter φ5 cm, height 90 cm) to recover the lolioside fraction. By concentrating and lyophilizing the recovered fraction, 3 g of a lolioside standard with a purity of 95% or more could be obtained.

[0044] (1.7) Preparation of Purified Lolioside Standard (2) 4 times the volume of water was added to 300 g of ryegrass seeds (flour) obtained by the method described in (1.1), and the water-soluble components were extracted. Then, the aqueous solution was recovered by filtration. 300 g of activated clay was added to the obtained solution, suspended, and filtered to remove the pigment components in the solution. The same solution was applied to an activated carbon column, and the oligosaccharide fraction was eluted with 30% (v / v) ethanol. The obtained oligosaccharide fraction was concentrated to 30 mL with an evaporator and subjected to a gel filtration column (carrier TOYOPEARL HW40S, Tosoh, column diameter φ5 cm, height 90 cm) to recover the lolioside fraction. By concentrating and lyophilizing the recovered fraction, 1.5 g of a lolioside standard with a purity of 95% or more could be obtained.

[0045] (1.8) Preparation of Purified Loliobiose Standard 3.2 g of the raffinose standard prepared by the above method was dissolved in 180 mL of water, and then 180 mL of 0.2 M hydrochloric acid - potassium chloride buffer (pH 1.9) was added, followed by acid hydrolysis at 60 °C for 22 hours. At the end of the reaction, sodium hydroxide was added to adjust the pH of the solution to neutral (pH 7.5), and then it was concentrated to 20 mL using an evaporator. The concentrated solution was subjected to a gel filtration column in the same manner as above to recover the raffinose disaccharide fraction. By concentrating and freeze-drying the recovered fraction, 1.1 g of a raffinose disaccharide standard with a purity of 95% or more was obtained.

[0046] (2) Results and Discussion (2.1) Palatability Test To verify the feed utilization of ryegrass seeds, the palatability of a commercially available feed and a ryegrass feed (granules) was compared. As a result, the intake during the feeding period was control feed = 332 ± 15 g, ryegrass feed (granules) = 304 ± 25 g (mean value ± standard error, n = 5), and no significant difference was observed. Considering that the tested chicks showed no difference between the two groups despite having been fed the control feed before the test, it is inferred that the palatability of ryegrass seeds is not bad.

[0047] (2.2) First Feeding Test A feeding trial was conducted using feed supplemented with LNB and ryegrass seeds (powder). To standardize the feed shape, ryegrass seeds (powder) crushed as described in (1.1) were used. The feeding results are shown in Table 2. There was no difference in feed intake among the groups, but the weight gain in the ryegrass group was significantly lower than that in the other groups. The feed efficiency in the ryegrass (powder) group was also significantly lower than that in the other groups. The ME content of the formulated ryegrass seeds was estimated to be 2.22 ± 0.06 kcal / g. It is considered that the reason for the low weight gain and feed efficiency in the ryegrass (powder) group was the insufficient energy in the feed. While the measured ME content of the control feed was 2.97 kcal / g, the ryegrass (powder) group was about 0.4 kcal / g lower at 2.59 kcal / g. Since there was no difference in feed intake among the groups, it was thought that by preparing the energy in the ryegrass feed to be equivalent to that of the control feed, feed performance comparable to that of the control group could be expected. For the LNB group, although no significant difference was observed compared to the control group, there was a tendency for better feed efficiency. The ileal amino acid digestibility of ryegrass seeds was generally lower than that of common feed ingredients, around 50%, and it was considered necessary to take amino acid digestibility into account in the formulation design.

Table 2

[0048] (2.3) Second feeding trial Feeding tests were conducted using diets supplemented with LNB and energy-adjusted ryegrass (milled). Ryegrass seeds were milled after crushing as described in (1.1) to ensure uniform shape and energy content. The ryegrass seeds were milled to remove fibrous components and had almost no change in the loliose content, so the loliose content was increased by 1.4 times. The feeding results are shown in Table 2. In the LNB group, there was a tendency for feed intake and weight gain to be improved compared to the control group, but this was not significant. On the other hand, in the ryegrass (milled) group, a significant improvement of 5% in weight gain was observed. No difference was observed in feed intake and feed efficiency between the groups. When preparing the ryegrass (milled) group, fats and oils as energy sources and some of the amino acids that were lacking were added to provide the same nutrients as the control group. As a result, the decrease in weight gain observed in the first feeding test was eliminated, and better weight gain than in the control group was confirmed. One reason for this is that the feed intake in the ryegrass (milled) group was higher than in the other groups, although there was no significant difference. In the first feeding test, the weight gain in the ryegrass-mixed feed group was also low, but the feed intake was comparable to the other groups, suggesting that the ryegrass itself is palatable. The ME contents of the control feed and the ryegrass-mixed feed were almost the same, at 3.09 and 3.05 kcal / g, respectively.

[0049] (2.4) Bifidobacteria cultivation test The turbidity of the culture supernatant for each strain after the end of cultivation is shown in Figure 1. Under the condition of adding loliose, the growth of five strains, JCM 6291T, JCM 8222T, JCM 1194T, JCM 19995T, and JCM 11165T, was observed. On the other hand, under the condition of adding loliose disaccharide, the growth of four strains, JCM 6291T, JCM 8222T, JCM 1194T, and JCM 19995T, was observed, but compared to the condition of adding loliose, JCM 6291T and JCM 8222T did not grow much.

[0050] [Example 2] (1) Method Test chickens: Broilers (Chunky) Test groups: 4 groups (group housing of 4 groups of 10 birds each × 4 groups = total of 160 birds) (1) Control ··········· Control feed for the second feeding test (2) Ryegrass seeds (ground) ··· Feed with ryegrass seeds for the second feeding test (3) Half amount of ryegrass seeds (ground) ··· Equal amount mixture of the above (1) and (2) (4) Group with 3% addition of fructooligosaccharide ··· Add 3% of reagent-grade fructooligosaccharide to the above (1) Feeding period: From 1 day old to 24 days old

[0051] (2) Results In broilers (Chunky), an equivalent significant weight gain effect was obtained compared to the control. Also, a significant weight gain effect was observed even when the proportion of ryegrass seeds (ground) was reduced by half. Furthermore, it was confirmed that the feed efficiency was improved by the mixing of ryegrass seeds (ground).

Table 3

Industrial applicability

[0052] The present invention can be used in industries related to feeds.

Claims

1. A feed composition comprising loriose or loriose disaccharide.

2. 2. The feed composition according to claim 1, which is for livestock or poultry.

3. 2. The feed composition according to claim 1, which is for chickens.

4. A method for producing a feed composition containing loriose or loriose disaccharide, comprising the following steps (1) to (3) and (5): (1) removing husks from seeds of one or more plants selected from the group consisting of plants of the genus Lolium and plants of the genus Festuca; (2) grinding the seeds obtained in step (1); (3) recovering the seed pulverized material passing through a sieve having an opening size of 0.5 to 2.0 mm from the seed pulverized material obtained in step (2); (5) A step of obtaining a feed composition using the crushed seeds recovered in step (3).

5. The method for producing a feed composition according to claim 4, further comprising: (4) recovering from the crushed seeds obtained in (3) any seed powder that does not pass through a sieve having an opening size of 100 to 200 μm; and (5) obtaining a feed composition using the crushed seeds recovered in (4).

6. A composition for growing microorganisms, comprising loriose or a loriose disaccharide.

7. 7. The composition for growing microorganisms according to claim 6, wherein the microorganism is a bifidobacterium.

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