JPS61247342A - Feed composition for ruminant - Google Patents

Abstract

PURPOSE:To provide a feed composition for ruminant having excellent rumen- bypassing efficiency and effective to remarkably improve the productivity of milch cow or fattening cattle by using N-acetyltryptophan or its salt as an essential component. CONSTITUTION:N-acetyltryptophan or its salt is added to a conventional feed composition for ruminant. The N-acetyltryptophan or its salt is passed through the rumen of e.g. a cow with little decomposition and absorbed in the body from the abomasum or intestine. It acts as tryptophan and increases the productivity of milch cow and beef cattle.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention provides N that is stable to the rumen contents of ruminants.
-Acetyl-L-tryptophan (hereinafter referred to as N-Ace
The present invention relates to the use of tyI-Trp) or a salt thereof by adding it to ruminant feed.

[Prior art and problems to be solved by the invention] Regarding tryptophan, we have already administered it to ruminants as tryptophan granules for rumen bypass.
We have previously filed a patent application regarding the application of L-Trp (hereinafter abbreviated as Trp) to lumen pipes and its unique functions. Generally, amino acids and proteins are fermented and metabolized by rumen microorganisms in the rumen, and most of them are decomposed by NH5 and CO2, and a portion of the NH3 is used as an N source by the rumen microorganisms. Microorganisms proliferate and serve as an important protein source for the host. Therefore, when amino acids are orally administered to ruminants, most of the amino acids are metabolically decomposed in the rumen, resulting in a large nutritional loss. Various lumen bypass techniques have been investigated in order to make more efficient use of the resources.

For example, a), Japanese Patent Publication No. 48-12785 b), Special Publication S-S-1057c) - Japanese Patent Publication No. 56-15495
6 and others, various proposals have been made in which biologically active substances are slurried and dispersed in molten hardened oils and fats, and then cooled and granulated. These granules contain a quaternary protective substance.
It is expected that it will disintegrate by the action of bile, water, etc. in the 12 denum, small intestine, etc. after the stomach, but because the time it remains in the gastrointestinal tract is short, it will not disintegrate sufficiently and biological The disadvantage is that most of the therapeutically active substances are excreted together with the protective substances. Also d), JP-A-58-1754
No. 49 attempts to improve the above-mentioned drawbacks by adding chitosan to fats and oils, but if there is too much chitosan, the durability within one lumen will be lost, and if there is too little, the above-mentioned patent a), It has the same drawbacks as b) and c), and is not expected to be effective. At present, there is no good product that exhibits both functions.

Generally speaking, increasing the durability within the rumen will result in insufficient solubility in the abomasum, and conversely, increasing the durability within the rumen will result in insufficient solubility in the abomasum. The result is that there is a shortage of To date, the rumen pi-guna method has not yet been achieved, which fully demonstrates the two functions of not being broken down in the lumen, but being reliably eluted from the abomasum and absorbed into the body. .

As another method for rumen pie gas, the example of the amino acid derivative Ashi, N-long chain acylmethionine has been proposed (e), Japanese Patent Publication No. 53-18429), but even in these cases, The durability within the lumen was not that good. However, if the amino acid is cheap like methionine or lysine, even if it is degraded to some extent in the rumen, it is conceivable to forcefully use it as a rumen gas amino acid, ignoring the large loss.0 However, in the case of Trp, on the other hand, there are special circumstances as follows, and it is obvious that it cannot be used with an insufficient sample of lumen pinox. That is, it has been estimated by RsJ, Carlson, et al. that Trp is metabolically degraded by microorganisms in the rumen to produce indole acetic acid (IAA), skatole (S) CA), and indole (END). Ap'plied Micr
obiology 27 (3) *540-548
(1974) Furthermore, the metabolites skatole and indole are harmful to ruminants, and the toxicity of skatole in particular may cause pulmonary edema (emphysema) in large amounts. They also revealed that oral administration of large amounts of Trp, such as 2007 or higher, causes pulmonary edema (emphysema) and death (
Am, J, Vst, Res 29αI 1983
~1.989 (1968)). Therefore, for these reasons, when administering Trp to cattle, unlike other amino acids,
Rumen Bypass It is an important requirement to maximize efficiency, and these cannot be achieved with the lumen bypass technology, which is an extension of the conventional technology. There is also concern that the harm caused by skatole may be greater than the effect if there is insufficient rumen pie dust captophane.

[Means and effects for solving the problem] We have focused on the special characteristics of TRP and have conducted intensive studies in search of a reliable lumen bypass technique, and as a result, we have arrived at the present invention.

That is, surprisingly, among the Trp derivatives, N-Ae@tyl-Trp, which is not a long-chain acyl derivative, stably and specifically acts against rumen microorganisms and the rumen fluid itself. It was found that it has excellent durability and the present invention was completed.On the other hand, N-propionyl-L-Trp is highly decomposable and produces sulfur and long-chain acyl-Trp.
It has been found that although the decomposition rate is somewhat improved with Trp (decamyl, lauroyl, myristoyl, valmitoyl), it has the major drawback of selectively increasing the generation of harmful skatole.

Since N-Acetyl-L-Trp is a powder and is expected to dissolve uniformly within the lumen at the dosage level, conventional granular lumen/? Il? Unlike sample samples, the residence time is also expected to be significantly shorter. In addition, N-Acetyl-L-Trp has excellent durability in the rumen, hardly produces skatole, which is extremely harmful to cows, and after being absorbed into the body, it is quickly metabolized to Trp and acts as Trp. The combination of these effects significantly improves the lumen binocular efficiency compared to conventional methods.This feature opens the door to the safe application of TRP to cattle for the first time. It is no exaggeration.

Furthermore, conventional rumen gas/JS technology, in which N-Acetyl-Trp or its salt is granulated and coated with various membrane materials, is slurried in a matrix of oils and fats and granulated. By applying this, it is possible to aim for even more efficient lumen supply A's.

The feed containing N-acetyl-Trp or its salt of the present invention is highly effective, for example, in improving the productivity of dairy cows, particularly in increasing the milk yield of high-temperature dairy cows. Or improving the productivity of feedlot cattle,
It has been found that this method is particularly effective in increasing wages for young fatteners.

For example, most of N-acetyl Trp or its salts passes through the cow's rumen without being broken down, and after reaching the abomasum or intestines, it is absorbed into the body and acts as Trp, effectively acting on the cow. It was found to increase the productivity of dairy and beef cattle.

N-acetyl Trp or its salt is for ruminants,
It can be fed as a powder orally with roughage or concentrate feed, dissolved in drinking water, or slurried in granules or fats and oils. It is characterized by the fact that it is free to choose any method that can be used to make money, and it has extremely high efficiency in lumen production.

N-Acetyl- passed through the rumen and reached the abomasum.
Regarding the behavior of Trp [Pharmacology and Treatment 8 (9) 309
3-3102 (1980)) and [Pharmacology and Treatment 8 (9
), 3103-3111 (1980) E, it has been proven that it functions almost equivalently as Trp. That is, N-Ace that has passed through the lumen
tyl-Trp exists stably even in the fourth organ, is absorbed into the blood below the small intestine, and is ultimately utilized effectively and exerts its effects, unlike Trp, by enzymes that are widely distributed throughout the body.

In addition, in terms of safety after absorption in the body, sufficient data has already been obtained, and it is recognized that it essentially acts almost equivalently to Trp. Examples that have been approved as a component of amino acid infusions and are already in practical use will be described below.

Example [1] Rumen fermentation of N-acetyl-L-Trp (I
Immediately after slaughter, the rumen contents were collected from three Holstein males separately using quadruple gauze to obtain sill-men fluid. A rumen fermentation test was conducted using this rumen fluid as follows. Add L-Trp solution N-Acetyl-L- dissolved in MB-9 buffer*IK to an Erlenmeyer flask in advance.
Take 4 portions of Trp solution separately and replace with N2.

A sample of 94 ml of rumen fluid was added to each Erlenmeyer flask while stirring well, and cultured at 40°C.

Sampling was performed at OHr, 8Hr, and an equal amount of 20
%) N-A after deproteinization with IJ dichloroic acid (TCA)
Cetyl-L-Trp and L-Trp were analyzed and quantified by HPLC.

(N-Acetyl-L-Trp, L-Trp M
B-9 solution preparation) N-Acetyl-L-Trp 15
Dissolve 0.811 Vi in MB-9, raise the volume to 25 mJ, and transfer this to a culture Erlenmeyer flask using a 41-hole pipette.

L-Trp 126, amp combined with MB-9 25m
1K) Collect the sample using a 41Le pipette and place it in a culture Erlenmeyer flask.

Acetyl-Trp exists as Acetyl-Trp or Trp in 93-97% after 8 hours of rumen fermentation, but only 1.0-8.8% of Trp remains.

This means that they are separated by more than 90 inches.

Reference Example 1 Using the same method as in Example 1, the following various compounds were subjected to rumen fermentation (in vitro) tests to estimate their durability in the rumen. Rumen fermentation was carried out using goat rumen fluid at 39°C for 8 hours. The results are shown in Table 2.

In the case of Acetyl-Trp, Acetyl-Trp
As a result, 91% of Trp remains, and 3% of Trp remains as a decomposition product.
The active ingredient is 8% and 94.8% when both are combined. Moreover, it is notable that no harmful skatole is produced.

On the other hand, compounds 1), 4), and 5) are samples that are extremely easily decomposed and cannot be used. Further, although the decomposition of compounds 2), 6), 7), 8), and 9) was suppressed, it was not as good as Acetyl-Trp. What is worse is that skatole, which is harmful to cattle as a decomposition product, is selectively produced in cows (11
．． 3-31.4%), it is therefore dangerous to directly administer these samples to cattle daily.

[Rumen fermentation of 23N-Acetyl-Trp was carried out in the same manner as in Example 1, except that one cow was used. The rumen contents of the cow immediately after slaughter were squeezed into a quadruple gauze to obtain rumen fluid. 11 in the rumen fluid was taken into a separating funnel, and after collection, it was left at room temperature for 2.5 hours and used for culture as it was without separating the fine feed pieces.

The changes over time in rumen fermentation are 0, , 3 after the start of culture.
, L-Trp, N-Acetyl-L-Trp were sampled at 8 hours, and some of them were analyzed by LC analysis after protein removal.
, indole acetic acid (IAA) was quantified.

A portion was extracted with chloroform to obtain indole (Ind).

Skatole (Ska) was quantified.

When compared in 8 hours, 50% of L-Trp was decomposed,
Indole acetic acid increased to 30.3%, indicating a sharp increase in skatole, which is harmful to cattle.

On the other hand, N-Acetyl-L-Trp has 13%
It has only been disassembled to a certain extent, and among them, the 5.6 section is L.
-Trp, and it is considered that there is virtually no decomposition. Particularly harmful skatole formation is caused by L-Tr
It is noteworthy that it is 1/30th of the average compared to p.

[3:] ] Stability of N-Acetyl-Tr in abomasal fluid and Holstein (male) (3 animals,
Collect the abomasal juices (referred to as A, &, C) and squeeze them into a quadruple gauze to obtain the abomasal juice. This abomasal juice is 481, N
Into two Erlenmeyer flasks with 2 substitutions, pre-adjusted Acetyl-Trp (2521n9/25mA!
MB-9) and Trp (189m9/25mgMg
-9) A 2M solution was added to separate Erlenmeyer flasks and changes over time were investigated at 39°C for 23 hours.

The results are summarized in Table 4.

Table 4: Stability of N-Aetyl-L-Trp and L-Trp in abomasal fluid N-Ac@t
yl-L-Trp.

It can be seen that it remains stable with almost no decomposition up to 3 hours. Even at 23 hours, 95 inches or more is stable.

L-Trp tends to be slightly more easily decomposed than N-Ac@tyl-L-Trp.

As a result of the above, lumen bypassed N-Aa@t
yl-L-Trp stably passes through the abomasum and is absorbed and utilized in the small intestine and beyond.

[4] Test with N-Acetyl-L-Trp dairy cows Using four Holstein dairy cows that have passed their peak lactation period, they were divided into a test group (2 cows) and a control group (3 cows) to test N-Acetyl-L-Trp.
An oral administration test of yl-L-Trp was conducted.

Preliminary period 10 days, test period 50 days L-Acetyl-L-Trp dosage = 6.0. ! i
'/D/1 cow, (feed 6I once in the morning with feed), Nutrient amount: 110 feed according to the Japanese feeding standard, roughage: concentrate feed ratio 50:50.

Investigation items: milk yield, milk fat percentage, milk protein, lactose.

Non-fat solid content results are shown in Table 5 and Figure 1. Both milk quality and reduction rate in the test plot exceeded the control plot, confirming the effect of the addition of N-Acetyl-L-Trp.

[5] Application to beef cattle Five 3-month-old male Holsteins were housed in groups in two groups, and their weights were measured up to 6 months of age and the amount of pay increase per head per day was compared. N-Ae@tyl-L-Tr in the test area
p61/D/head was given once daily in the morning along with feeding.

The feed provided was 11096 feed, which is the Japanese feeding standard, and the ratio of roughage to concentrate feed was 50:50.

Feed DM Atashi's EP included was 16 yen.

The results are summarized in Table 6. In the DC comparison, the test area was 6.8 ry more expensive.

[6] Acetyl-Trp powder 900. Finely powdered NaCl2101' is thoroughly mixed in F and used as a granulation raw material, and the mixture is processed using a centrifugal granulation coating machine (CF-360) (
It was granulated using Freund Sangyo KK). Nonno in the nucleus? Reru 101500 g (20-24 Mesh)
81 I (PC ethanol solution) was used as the binder.

Granules with a particle size of 1-2% 3590! i was obtained.

At the time of 3.0 of this granulation, beef tallow: AEA* is used as a coating material.
1=1:1 (wt ratio) was dissolved in chloroform to make a 5.0% solution, and coated to 3Q wt% based on the weight of the granules using the above coating machine.

Product composition: Acetyl-Trp 19.4% NaC245
, 3% RPC 2.0% Beef tallow: AgA 22.5%*1 AEA is an enteric coating material manufactured by Sankyo Pharmaceutical [Digestion test] This granular coating sample of Acetyl-Trp was applied to dairy cows. 15th floor one day a week! S'am was fed with feed every morning, and whole wings were collected during the last two days of the test period.
When testing the state of digestion of p1e, no particulate matter was detected in the feces, and analysis of the feces revealed that Acet
Since yl-Trp was not detected, N-Acetyl
It is estimated that -L-Trp was effectively digested and absorbed within the gastrointestinal tract.

Regarding this rumen biomass sample, the durability in the rumen was determined using the nylon bag method, and the dissolution rate was 1.
It was 3.2% (20Hra).

Among the further eluted N-Acetyl-L-Trp,
Since the decomposition rate is 5.0% (8 Hr) (Reference Example 1), the true decomposition rate of N-Acetyl-L-Trp is about 0.7%, which is almost completely the lumen bypass sample. I can say that there is.

Claims (1)

[Claims] Ruminant feed containing N-acetyltryptophan or its salt