JP2002078456A - Feed for pig breeding and method for breeding pig - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a feed for pig breeding, capable of preventing pigs from suffering by harmful bacteria infection caused via the intestinal canal regardless of the kind of bacterial species/bacterial strains. SOLUTION: This feed for pig breeding is characterized by being formulated with a hydrolyzate obtained by decomposing copra meal or palm kernel meal using an acid or enzyme.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a feed for preventing harmful bacterial infection of pigs and a method of breeding pigs, and more particularly, to inhibiting the infection of harmful bacteria occurring via the intestinal tract, The present invention relates to a feed and pig breeding method for improving health, productivity, and safety of the livestock and poultry products.

[0002]

2. Description of the Related Art Infection with pathogenic bacteria such as pathogenic Escherichia coli, Salmonella, Campylobacter and the like causes a decrease in productivity of pigs, and also poses a serious problem in terms of food safety of the livestock. In particular, stress caused by weaning of piglets, change of feed, and movement of pig bunches lead to a decrease in disease resistance, causing pathogenic bacterial infections and diarrhea, which is a major problem.

Until now, antibiotics have been used to prevent harmful bacterial infections in pigs. However, although the use of antibiotics has a certain effect of preventing harmful bacterial infections, it cannot be used positively due to the problem that antibiotics remain in meat and the emergence of resistant bacteria.

Also, lactic acid bacteria, butyric acid bacteria, bifidobacteria, C
It has been reported that by adding a live bacterial agent such as an E (Competitive Exclusion) preparation to a feed, useful bacteria are provided, and the effect of inhibiting harmful bacteria from colonizing the intestinal tract has been reported.
However, the useful bacteria used for such a probiotic agent have a difference in intestinal colonization depending on the animal species, and there is a problem that useful bacteria may not be sufficiently established even in the same animal species. Remaining.

On the other hand, harmful bacterial infection that occurs via the intestinal tract is caused by the adhesion of harmful bacteria to the intestinal tract. Therefore, it is necessary to add a substance that inhibits the adherence to the intestinal tract to the feed to prevent the infection. Is attracting attention. It has been reported that the adhesion of the harmful bacteria to the intestinal tract occurs by recognizing the sugars of glycoproteins or glycolipids present on the surface of the intestinal tract.

[0006]

It is known that the adhesion factors of the pathogenic bacteria to the intestinal tract are different depending on the strains and species (Osamu Fujita et al .; Bioscience and Industry 55: 181). -186, 1997
Year). For example, it is known that pathogenic bacteria having a type 1 adhesin such as Salmonella, Escherichia coli, and Shigella recognize and attach mannose on the surface of the intestinal tract. On the other hand, it has been reported that Escherichia coli having a K99 adhesin, which is frequently detected in piglet colibacillosis, recognizes and adheres to saccharides other than mannose of glycolipids.

[0007] Therefore, attempts have been made to prevent the transmission of harmful bacteria having type 1 adhesin by adding mannose and mannooligosaccharides to feeds. At present, not only is it not effective against harmful bacteria having type 1 adhesin, but also does not sufficiently inhibit adhesion to the intestinal tract.

[0008] An object of the present invention is to provide a feed for swine raising and a method of breeding pigs for preventing infection of harmful bacteria occurring via the intestinal tract.

[0009]

Means for Solving the Problems The present inventors have conducted intensive studies in order to solve such problems, and as a result, have obtained hydrolysates obtained by partially hydrolyzing copra meal or palm kernel meal with acids or enzymes. The inventors have found that the degradation product is not affected by the species and strain of the harmful bacteria and significantly inhibit the adhesion of the harmful bacteria to the intestinal tract, thereby completing the present invention.

That is, a first aspect of the present invention is to provide a feed for swine raising, which comprises a hydrolyzate obtained by decomposing copra meal or palm kernel meal with an acid or an enzyme. The second is a gist of a pig breeding method using the above feed.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail. Copra meal is a pulverized residue obtained by extracting coconut oil from copra, a coconut oil raw material obtained by drying the core meat of coconut fruit, and contains henancellulose such as mannan, glycolipids, and glycoproteins. It is contained. Copra meal, which is a raw material of the feed for swine raising of the present invention, is not particularly limited in its origin and production method as long as it is produced in a usual coconut oil production process.

[0012] Palm kernel meal is a pulverized residue obtained by extracting palm kernel oil from palm kernel of oil palm seeds. Like copra meal, hemicellulose such as mannan, glycolipids and glycoproteins are used. Etc. are contained. The palm kernel meal used as a raw material for the feed for swine raising of the present invention is not particularly limited in its origin, production method, etc., as long as it is produced in a usual palm kernel oil production process.

The enzyme that acts on copra meal or palm kernel meal is not particularly limited as long as it acts on hemicellulose and releases mannose, and examples thereof include hemicellulases such as mannanase (mannase) and mannosidase. As for the origin of such enzymes,
Bacillus subtilis, filamentous fungi (Aspergillus aculeatus,
A. awamori, A .; niger, A .; usami
i, Humicola insolens, Trich
orderma harzianum, T.M. koning
i, T. Longibrachiatum, T .; vir
ide), Basidiomycetes (Corticium, Pycnop)
orus coccineus).
Spergillus-derived enzymes are preferred. Among them, mannanase derived from Aspergillus niger is particularly preferred.

[0014] These hemicellulases are produced in the culture supernatant or cells of the above strains, and any fraction containing these hemicellulases may be used. If necessary, fractions containing these hemicellulases containing purified or partially purified conventional methods may be used.

Also, Cellulosin HC100, Cellulosin HC, Cellulosin TP25, Cellulosin GM5 (all manufactured by Hankyu Bio Industries Co., Ltd.), Sumiteam A
Commercially available enzymes such as C, Sumiteam AC-L, Sumiteam ACH (above manufactured by Nippon Chemical Industry Co., Ltd.) and Gamanase (manufactured by Novo Nordisk Industries) can also be used.

In the present invention, a hydrolyzate can also be obtained by allowing an acid such as sulfuric acid or hydrochloric acid to act on copra meal or palm kernel meal. The concentration of the acid used is preferably 20 to 90% by volume, more preferably 60 to 80% by volume in the case of sulfuric acid. The hydrolysis conditions are 80
~ 121 ° C is preferred.

In the present invention, the hydrolyzate of copra meal or palm kernel meal can be obtained by allowing the above enzyme or acid to act on copra meal or palm kernel meal. The hydrolyzate of copra meal or palm kernel meal can change the concentration of mannose and other active ingredients depending on the acid concentration, the amount of enzyme added, and the reaction time. Has a mannose content of 0.5% or more,
Preferably, the reaction conditions are such that the mannose content can be 5% or more.

The concentration of the hydrolyzate of copra meal or palm kernel meal added to the feed varies depending on the age of the pig, but is 0.001 to 5% by weight, preferably 0.005% by weight.
It may be 〜1% by weight.

[0019]

The hydrolyzate of copra meal or palm kernel meal according to the present invention contains mannose as a main component, but has a significantly higher effect of preventing harmful bacterial infection than the case where mannose or mannooligosaccharide is used alone. Although this factor is not clear, the hydrolyzate of copra meal or palm kernel meal releases, along with mannose, various substances such as glycolipid-derived saccharides and glycolipids that are easily dissolved by the action of enzymes. Therefore, it is presumed that these substances become receptors or receptor analogs of various harmful bacterial adhesion factors and can prevent harmful bacterial infection without being affected by bacterial species and strains.

[0020]

The present invention will be described below in detail with reference to examples. Reference Example 1 [Preparation of copra meal hydrolyzate] Cellulosin GM5 (mannanase manufactured by Hankyu Bioindustry Co., Ltd., titer 10,000 units / g)
25 g is suspended in 100 mL of water, and 100 g of copra meal is suspended.
(A fat content of 10% by weight and a water content of 7.2% by weight), and then left at 60 ° C. for 18 hours. After completion of the reaction, a vacuum dryer (Vaccum Dryi, manufactured by Yamato Corporation)
ng Oven DP32) at 80 ° C. for 24 hours under vacuum to obtain a copramyl hydrolyzate. The analysis of the sugar component in the target product was performed by high performance liquid column chromatography. Analytical column is Aminex H manufactured by Bio-Rad
PX-87P was used. The column temperature was 85 ° C and the flow rate was 0.
Elution was performed with water at 6 ml / min. The sugar was detected using a differential refractometer, and the mannose content was determined from the quantitative value of the standard product. As a result of analyzing the powder after the above reaction,
10.3 g of mannose had accumulated in 100 g.
The water content was 9.6% by weight.

Reference Example 2 [Preparation of palm kernel meal hydrolyzate] Cellulosin GM5 (mannanase manufactured by Hankyu Bio-Industry Co., Ltd., titer 10,000 units / g)
25 g was suspended in 100 mL of water, sprayed uniformly on palm kernel meal, and then left at 60 ° C. for 18 hours. After the reaction, vacuum dryer (Vac, manufactured by Yamato Corporation)
Cum Drying Oven DP32) was vacuum dried at 80 ° C. for 24 hours to obtain a hydrolyzate of palm kernel meal. The analysis of the sugar component in the target product was performed by high performance liquid column chromatography. The analytical column used was Aminex HPX-87P manufactured by Bio-Rad. Column temperature is 85 ° C,
Elution was carried out with water at a flow rate of 0.6 ml / min.
The sugar was detected using a differential refractometer, and the mannose content was determined from the quantitative value of the standard product. As a result of analyzing the powder after the above reaction, 18.3 g of mannose was accumulated in 100 g. The water content was 8.4% by weight.

Example 1 Pathogen Inhibition Test Intestinal epithelial cells were removed from piglets (20-day-old, large Yorkshire) and cultured in a CO ₂ incubator for 12 hours using an RPMI 1640 medium containing 10% fetal bovine serum, and the monolayer was cultured. Formed. After washing with RPMI 1640 medium, RP
5 × 10 ⁸ swine-derived pathogenic Escherichia coli O78 (K88 adhesin possession) was added to the copra meal hydrolyzate and palm kernel meal hydrolyzate at the target concentration in MI1640 medium.
The mixture was added to the above monolayer of intestinal epithelial cells and incubated at 37 ° C. for 30 minutes in a CO ₂ incubator. Then, by removing the unattached bacteria, and washed three times with RPMI1640 medium,
Bacteria adsorbed to intestinal epithelial cells non-specifically were removed.

The cells were observed with an electron microscope, the number of bacteria adhering to 50 cells was counted, and the average value was taken as the number of bacteria adhering to one cell. Adhesion inhibition rate X
(%) Is the number of bacteria attached to one cell in the test group, Y is the number of bacteria attached to one cell in the control (no addition) group, and X = (1-Y / Z) × 100. Further, for comparison, a test was conducted on a sample to which mannose was added to be 0.1% (Comparative Example 1).

The results obtained are shown in Table 1. Table 1 is a table showing the ratio (%) in which the adhesion of pathogenic Escherichia coli was inhibited in the pig-derived pathogenic Escherichia coli adhesion inhibition test. As shown in Table 1, as compared with the control and the comparative example, a significantly higher adhesion inhibitory effect was confirmed for the copramyl hydrolyzate and the palm kernel meal hydrolyzate against K88-containing pathogenic Escherichia coli showing adhesion that does not involve mannose. Was done.

[0025]

[Table 1]

Example 2 Pathogenic Escherichia coli (O10
1) Infection suppression test Piglets weaned at the age of 20 days (Large Yorkshire breed) were divided into 15 pigs so that the average body weight and gender ratio became almost equal, and 0.1% by weight of the copra meal hydrolyzate prepared in Reference Example 1 %
The supplemented feed supplemented with was continuously provided. On the 7th day after feeding, 2.0 × 10
5 mL of a bacterial solution containing ⁶ cells / mL was orally administered by gavage, and the growth rate and the incidence of diarrhea on days 3, 7, and 14 after administration of Escherichia coli were examined. For comparison, Escherichia coli bactericidal test was similarly conducted using a feed (Comparative Example 2) containing 0.1% by weight of mannose (manufactured by Ishizu Pharmaceutical Co., Ltd.).

Tables 2 and 3 show the obtained results. Table 2 shows the breeding rate (survival rate) of pigs in the test for controlling E. coli (O101) infection using pigs, and Table 3 shows the incidence of diarrhea in pigs in the test for controlling E. coli (O101) infection using pigs. Is shown. Table 2 shows that, in the section where the copra meal enzyme digest was added, the growth rate was higher than that in the case where no enzyme was added and in Comparative Example 2, and the effect of preventing infection with pathogenic Escherichia coli was confirmed. Table 3 shows that the incidence of diarrhea was not added and Comparative Example 2 was added.
The results show that the copramyl hydrolyzate effectively inhibited the adhesion of pathogenic Escherichia coli to the intestine from the fact that the value was significantly lower than that of.

[0028]

[Table 2]

[0029]

[Table 3]

Example 3 Pathogenic Escherichia coli (O14
1) Infection control test Piglets (large Yorkshire breed) weaned at the age of 20 days were divided into 15 pigs so that the average body weight and gender ratio were almost equal, and the palm kernel meal hydrolyzate prepared in Reference Example 2 was used. Formula feed supplemented with 1% by weight was continuously provided. After feeding, 7
On the day, pathogenic Escherichia coli O141 (having type 1 adhesin)
The 2.0 × the bacterial solution 5mL oral gavage containing 10 ⁶ cells / mL, 3 days after E.coli administration, day 7 were examined raising rate and diarrhea incidence of 14 days. For comparison, Escherichia coli bactericidal test was similarly performed using a feed (Comparative Example 3) containing 0.1% by weight of mannose (manufactured by Ishizu Pharmaceutical Co., Ltd.).

Tables 4 and 5 show the obtained results. Table 4 shows the breeding rate (survival rate) of the pigs in the test for suppressing the pathogenic Escherichia coli (O141) infection using pigs, and Table 5 shows the incidence of diarrhea in the pigs in the test for suppressing the pathogenic Escherichia coli (O141) infection using pigs. Is shown. As shown in Table 4, in the palm kernel meal hydrolyzate-added section, no addition and comparative example 3
It was a high growth rate. Further, as shown in Table 5, the incidence of diarrhea was significantly lower than that of Comparative Example 3 without addition, and it was found that the adhesion of pathogenic Escherichia coli to the intestine was effectively inhibited by the palm kernel meal hydrolyzate.

[0032]

[Table 4]

[0033]

[Table 5]

Example 4 Pathogenic Escherichia coli G12 Using Piglets
53 strains (K88 possession) artificial infection test Using 20 piglets (malted male and female) immediately after weaning (23-27 days old), acclimated to the breeding environment 10 days before the start of the test, based on the weight of the test start day , Each group was divided into 4 groups by a stratified random method so that the average body weight of each group was approximately equal, and housed in each pig group in a closed pig breeding room adjusted to 27 ° C. . During the acclimatization period, the standard feed SD for piglet milk
S No.1 (manufactured by Japan Combined Feed Co., Ltd.) was fed ad libitum, and drinking water was allowed to be freely ingested from a tap water supply.

The test substance and the control substance were added to the standard feed SDS No. 1 for the initial period of pig milk artificial milk so as to have the respective additive concentrations shown in Table 6 below. They were fed continuously for 14 days from the start of the test to the end of the test. In addition, a dry yeast cell wall (manufactured by Alltech, INC., Sold by Bussan Biotech Co., Ltd., trade name: Biomoss) was used as a control, and the control group was not added.

[0036]

[Table 6]

As the strain used for the artificial infection, a strain obtained by imparting streptomycin resistance to BSE4004 distributed by the Association of Animal Biologicals was used. This strain was resistant to 1,000 μg of streptomycin titer / ml, and it was confirmed that it had K88 by agglutination reaction using K88 serum. Bacterial liquid administration was performed twice, 7 days after the start of the administration of the test substance and 4 days after that. The number of bacteria administered per animal was 2.1 × 10
¹⁰ CFU / 2 ml, and the second time was 1.0 × 10 ¹⁰ CFU / 2 ml.

During the test period, the general condition, the presence or absence of diarrhea, the weight measurement, and the amount of feed intake were measured. In addition, about diarrhea symptom, 0: normal stool, 1: loose stool (softer than normal stool, but presents a stick shape), 2: muddy stool (liquid, but less water than watery stool ) 3: Watery stools (liquid and squirted vigorously from the anus) were evaluated in four stages. Table 7 summarizes the changes in diarrhea symptoms.

[0039]

[Table 7]

From these results, it was found that diarrhea symptoms were improved in the copramyl enzyme-decomposed product-added group as compared with the control group and the dry yeast cell wall-added group. Clearly, it can be eased.

Table 8 summarizes changes in body weight of the test animals. It is clear that the group fed with the test substance had a higher body weight gain than the control group and the biomoss-added group.

[0042]

[Table 8]

Table 9 summarizes the feed demand rates.
The group fed the copra meal enzyme hydrolyzate had a clearly lower feed requirement than the control group and the dry yeast cell wall-added group, indicating that the feeding of the copra meal enzyme hydrolyzate was effective.

[0044]

[Table 9]

[0045]

EFFECT OF THE INVENTION By the feed of the present invention, the species of harmful bacteria
It is possible to inhibit the adhesion of harmful bacteria to the intestinal tract without being affected by the strain, and to prevent infection of pigs with harmful bacteria.

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Claims (2)

[Claims] 1. A feed for swine, comprising a hydrolyzate obtained by decomposing copra meal or palm kernel meal with an acid or an enzyme. 2. A method for breeding pigs, comprising breeding pigs with the feed for swine raising according to claim 1.