JPH0445751A - Feed for animal - Google Patents

Abstract

PURPOSE:To obtain the title feed containing eicosapentaenoic acid-bonded phospholipid consisting essentially of phosphatidylethanolamine, etc., as an active ingredient and capable of bringing out lowering of blood pressure, lowering of weight of fat structure and lowering of liver triglyceride and plasma lipid. CONSTITUTION:The aimed feed containing eicosapentaenoic acid-bonded phospholipid consisting essentially of phosphatidylethanolamine and/or phosphatidyl glycerol and/or a microorganism or algae containing the above-mentioned phospholipid as an active ingredient. Furthermore, the above-mentioned microorganism is preferably the genus Pseudomonas, Alteromonas, Shewanella, Deleya, Alteromarinus, Pasteurella or Vibrio.

Description

[Detailed description of the invention] (Industrial application field) The present invention relates to a novel animal feed.

(Prior art) Today, typical adult diseases such as arteriosclerotic diseases, thrombotic diseases, hyperlipidemia, obesity, hypertension, and diabetes are all caused by disturbances in normal lipid metabolism. However, such diseases have recently been rapidly increasing not only in human society but also in vented animals, and have become a big problem for human owners as well.

Furthermore, the meat of large animals such as cows and sheep for meat, as well as the meat of chickens and eggs, are high in fat (e.g., neutral fat and cholesterol), leading to problems in terms of excessive calorie intake and feeding. There is. Several methods have been proposed to treat and prevent these diseases, as well as to improve meat and egg quality, but none of them can be said to be fully satisfactory in terms of their effectiveness, and safer methods have been proposed. Demand for effective feed is increasing. Recently, eicosapentaenoic acid (hereinafter referred to as EPA) has been attracting attention due to its high safety.The chemical forms of EPA that have been developed so far have mainly been in the form of ethyl ester or triglyceride; Attempts have also been made to produce EPA-bound phospholipids by chemical synthesis, since the chemical form of EPA-bound phospholipids produces rapid effects at a dose of 1/10 or less (Japanese Unexamined Patent Application Publication No. 1983-1996). -264422, JP-A-58-38215).

However, when these chemical synthesis methods are used, contamination with contaminants that are clearly harmful to growth cannot be avoided, and furthermore, the reaction efficiency is low, resulting in high manufacturing and purification costs, making them far from practical.

(Problems to be Solved by the Invention) An object of the present invention is to provide a new, more effective and safer animal feed in large quantities and at low cost.

(Means for Solving the Problem) As a result of intensive research on animal feed, the present inventors have discovered that EP
The following findings were obtained regarding the lipids of microorganisms capable of producing A. That is, ■ &t1 of microorganisms capable of producing EPA.
) content is high at 6-12% on dry basis, of which more than 90% is phosphatidylethanolamine (hereinafter referred to as PE) and phosphatidylglycerol (hereinafter referred to as PG).
■The fatty acid composition of the total lipids shows that EPA is high at 25-40%, and that it is the only highly unsaturated fatty acid that contains -EPA. High purity phospholipids can be easily obtained by fractionation by fusing and column separation. Furthermore, the EPA used here
The growth rate of microorganisms capable of producing microorganisms is significantly faster than that of ordinary microorganisms, and a large amount of microorganisms can be obtained in an extremely short period of time. Furthermore, the lipids and adipose tissue weight in the blood and liver of animals to which EPA-bound phospholipids containing PE and/or PC as main components and/or microorganisms and algae containing these extracted from the above-mentioned bacterial cells were administered were effectively reduced. It can be lowered to

The present invention was completed based on the above findings, and
The present invention relates to a novel animal feed containing, as an active ingredient, an EPA-bound phospholipid containing E and/or PC as a main component, and/or microorganisms or algae containing the same.

Microorganisms capable of producing EPA-bound phospholipids that can be used in the present invention are particularly suitable for microorganisms of the genus, as long as they have a high EP^ content.
Although not limited to species or strains, usually
Shewanella
), the genus Alteromarina, the genus Pasteurella, and the genus Bibliobum, which have the ability to produce EPA. Using microorganisms. These microorganisms are described in European Patent Application No. 8731)372.4 and Japanese Patent Application No. 8731-372.4, which the present inventors previously proposed as microorganisms for EPA production.
49929, patent application No. 62-49930, patent application No. 62-4
9931. Patent application 1986-156693 and patent application 1986-
273200 describes its properties in detail.

For i[l[, which contains 8-way EPA-bound phospholipids,
Euglena gracilis■■pressure■ Red acilis) A
Examples include TCC12893, Chlorella minutissima type sockeye salmon ainutissim (transformation), Marchantia polymorpha (Marchantia shark n embryo), and the like.

In carrying out the present invention, microorganisms and algae capable of producing EPA-bound phospholipids are cultured in a conventional manner, for example, in a medium to obtain a national body, which is collected by centrifugation and dried to produce microorganisms and G*. Get the powder. As the culture medium at this time, for example, a culture medium having a composition shown in Table 1 below can be used.

Table 1 (composition of medium) The bacterial cell phospholipid fraction can be extracted from this microorganism or algae powder. There is no particular limitation on the method, but for example, lipid extraction is carried out using molten molten oil, and after removing f@,
By performing acetone fractionation and collecting unnecessary components in acetone, a crude fraction of phospholipids can be obtained. For Aritsuki solubles, ordinary solvents that dissolve phospholipids may be used alone or in combination, such as chloroform-methanol, n
Examples include -hexane-ethanol-water system, chloroform-hexane system, chloroform-ether system, chloroform-ethanol system, and the like.

Further, as the solvent used for fractionation, an organic solvent that is normally used for fractionation of lipids can be used.

Examples include acetone, acetic acid, chloroform, and ether. Moreover, MgCLt, calcium, etc. may be added to the solvent, and the fractionation may be performed in the presence of the solvent, and heating and cooling may be performed as necessary.

The composition of the phospholipids obtained in this way is analyzed by TLC (II layer chromatography), and then saponified and decomposed by conventional methods, methyl esterified, and analyzed by GLC (gas chromatography) to determine the constituent fat #. The composition can be determined.

Phospholipid composition obtained by the above method (EPA-P
L) contains phosphatidylethanolamine and phosphatidylglycerol as main components, and can be easily fractionated using a DEAE cellulose column with a chloroform-methanol-based elution solvent. Its constituent fatty acids are E
PA, oleic acid, palmitoleic acid, palmitic acid,
It was heated with myristic acid etc.

The phospholipid fraction of microorganisms and algae having the ability to produce EPA-bound phospholipids and/or the bacterial cells and algal cells obtained as described above are used as animal feed. The dosage at this time is not particularly limited, but for example, it is approximately 1 ■/one body weight/day to Ig/one body weight/day in terms of EPA-bound phospholipid, and when administered in the form of a hedron or algae. The same effect can be obtained with about 20 times the amount of the animal feed of the present invention. There is no particular limitation on the form of the animal feed of the present invention. Forms include granules, tablets, suspensions/emulsions, powders, tablets, and syrups. It may also contain biologically and pharmaceutically acceptable additives.

The animal feed of the present invention can be used not only for the prevention and treatment of diseases in animals such as livestock and beds, but also in humans.
It can be widely used as a health food, or by administering it to chickens to produce low-cholesterol eggs or eggs containing EP.

w, Table 2 (composition of feed) Example I Microorganism having EPA-producing ability (Alteromonas huu)
'J 77 'J engineering:/su5CRC2874 (wl,
Technique 1) i1 Jyo No. 9158)) was cultured in a medium containing 1% peptone, 0.5% yeast extract, and 5% NaCll and adjusted to pH 7, and the bacterial cells were collected by centrifugation, washed,
After drying, a microorganism powder was obtained. Extract 100g of microorganism powder with chloroform:methanol-2=1 mixed solvent and extract 8g
Extracted lipids were obtained. Acetone 4 is added to the total lipid after this solvent removal.
00 was added, stirred while cooling, and acetone-insoluble matter was collected by filtration. When this acetone-insoluble matter was removed from the solvent, about 7 g of a phospholipid composition was obtained. As a result of TLC analysis, the composition of the obtained phospholipid was found to be mainly composed of phosphatidylethanolamine (PE) and phosphatidylglycerol (PG). After dissolving these in a small amount of chloroform, a DEAE cellulose column (33φX4Qcs ) and eluted with increasing methanol content sequentially starting with chloroform. PE and PC fractions were collected while checking with TLC, and the solvent was distilled off. Approximately 5 g and 2 g of PE and PG were recovered, respectively. The constituent fatty acids were EPA, oleic acid, valmitic acid, palmitoleic acid, myristic acid, etc.

The PE and PC were mixed into the experimental animal feed shown in Table 2 and used.

As a control feed, a feed containing 10% soybean lecithin instead of EPA phosphorus was used. E above
PA phospholipid diet or soy lecithin diet was administered to 5HR-3P rats (III stroke-prone rats, male, body weight 236 g).
> 5g per day was administered, and further 13g of normal feed (SP food, manufactured by Funabashi Co., Ltd.) was administered per day.

The animals were raised in this way for 4 weeks, during which time the tail artery blood pressure was measured every week.

Four weeks after administration, the animals were dissected and the weight of adipose tissue, the amount of triglyceride in the liver, and the plasma lipid were measured. Triglycerides in the liver are chloroform:methanol-2:
Total lipids were extracted with a mixed solvent and then measured using the acetylacetone method. Plasma total cholesterol was measured by the Zulkowski method, and VLDL and chylomicron were fractionated into each lipoprotein by ultracentrifugation, and then the amount of cholesterol was measured by the same method. Plasma triglyceride and phosphorus II were measured by the acetylacetone method and phosphorus ashing method, respectively.

The results obtained in this way are shown in Figure 1 and Tables 3 to 5.
The numbers in parentheses in Tables 3 to 5 indicate the rate of decrease (%) when the control value is 100%.

Table 3 (Decrease in fat & lv & weight) Table 4 (Decrease in liver triglyceride) Table 5 (Decrease in plasma lipid) (Effects of the invention)
EPA-containing phospholipids containing phosphatidylethanolamine and/or phosphatidylglycerol as main components can be obtained in high yield, and the phospholipid composition can be
When administered to R-3P la-/), a decrease in blood pressure, a significant decrease in fat & ll type weight, and a decrease in liver triglycerides and plasma lipids were observed. It has been shown that the microbial cells and/or the bacterial cells and algal cells can be used as an effective and inexpensive new animal feed.

4,

[Brief explanation of drawings]

FIG. 1 is a diagram showing the relationship between feed used in Examples and blood pressure fluctuations.

Claims (3)

[Claims] (1) For animals, characterized by containing as an active ingredient an eicosapentaenoic acid-binding phospholipid containing phosphatidylethanolamine and/or phosphatidylglycerol, and/or a microorganism or algae containing the same. feed. (2) The microorganism described in (1) is Pseudomonas (Ps
eudomonas) genus, Alteromonas (￥Alte
romonas￥), Shewanella (￥Shewane)
The genus Deleya, Deleya, Alteromarinus, Pasteurella, or Vibrio according to claim 1 Animal feed. (3) The algae described in (1) is Chlorella (¥Chlor
The animal feed according to claim 1, which belongs to the genus Ella, Euglena, or Marchantia.