JP2006325587A - COMPOSITION, FEED ADDITIVE AND FEED EACH DESIGNED TO CONTROL CAMPYLOBACTER, AND USING METHOD OF beta-1,4-MANNOBIOSE-CONTAINING COMPOSITION - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a composition, feed additive and feed each designed to control campylobacter, safely and easily suppressing remaining of campylobacter bacteria in animal intestine and efficiently discharging campylobacter bacteria to the outside of a body, and to provide a using method of the composition, the feed additive, and the feed. <P>SOLUTION: The composition, the feed additive and the feed each designed to control campylobacter contain β-1,4-mannobiose. The using method of the β-containing composition is also provided. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a composition for combating Campylobacter, a feed additive and a feed, and a method for using a β-1,4-mannobiose-containing composition.

  Conventionally, it has been known that palm kernel meal, copra meal, guar meal, and the like contain abundant mannan containing mannose as a constituent sugar. Various methods for producing saccharides have been proposed.

  In addition, feed supplemented with such mannoses is known to have the effect of eradicating Salmonella, which is a harmful bacterium, in the animal intestine and discharging it outside the body (Salmonella colonization suppression effect). Various techniques using this effect have also been proposed (Non-Patent Document 1).

  For example, feed containing manno-oligosaccharides, which are mainly composed of mannobiose and mannotriose and mixed with monosaccharides, obtained by enzymatic degradation of mannan-containing materials such as guar meal and copra meal can improve the quality of chicken eggs. It has been reported that manno-oligosaccharides may be useful for preventing colonization of Salmonella in the gut of livestock (Patent Document 1).

  In addition, feed containing mannose such as mannan enzyme and / or acid hydrolyzate obtained from mannobiose, methyl-α-mannoside, manno-oligosaccharide, guar gum, locust bean gum or yeast is a harmful bacteria such as Salmonella. It is reported that it is useful for infection prevention (patent document 2).

  Furthermore, a livestock feed containing a mannose-based polysaccharide that is mainly composed of polysaccharides having a mannose repeat unit of 40 to 100 (30 to 80%) and oligosaccharides (5 to 30%) is a salmonella livestock intestine. It has been reported that it is effective in preventing fixing in the inside (Patent Document 3).

  In addition, feed containing a mannan-rich raw material such as palm kernel meal or guar meal mixed with manno-oligosaccharide and a method for producing mannose and / or manno-oligosaccharide using copra meal with the enzyme are also reported. (Patent Documents 4 to 6).

Furthermore, mannose is easily utilized by intestinal bacteria, or digested, absorbed and excreted by the livestock itself, and mannose is easily decomposed when a live fungus is administered. It has been reported that mannobiose, which is a sugar, can effectively exert the effect of suppressing Salmonella colonization compared with mannose (Patent Document 7).
Poultly Science, 68, p.1357, 1989 JP-A-8-38064 JP-A-7-236429 JP-A-8-173055 WO95 / 17103 pamphlet International Publication No. 99/08544 Pamphlet JP 2001-231591 A International Publication No. 2004/048587 Pamphlet

  By the way, Campylobacter is known as a causative agent of food poisoning frequently occurring along with Salmonella, Vibrio parahaemolyticus, Staphylococcus aureus and the like. For example, according to the state of occurrence of food poisoning by pathogenic substances in 2003 (Ministry of Health, Labor and Welfare statistics), the number of cases of food poisoning is most frequently caused by Campylobacter bacteria. The causative food is most often related to chicken, about 80%, and is often caused by eating chicken that is not sufficiently heated. Campylobacter itself has been isolated at a high frequency in chicken and poultry processing facilities, and it has been reported that it was detected from about 70% of commercially available meat.

  As an effective means for preventing the contamination of meat by such Campylobacter bacteria, it is conceivable to prevent the contamination of the poultry itself such as broilers, but a safe and simple means has not yet been found.

  An object of the present invention is to safely and simply suppress the establishment of Campylobacter bacteria in the animal intestine and efficiently discharge Campylobacter bacteria to the outside of the body. It is in providing the additive for feed, feed, and these usage methods.

  As a result of intensive studies, the present inventor found that β-1,4-mannobiose has a Campylobacter recognition activity and is less likely to be assimilated by bacteria in the intestines of livestock, and suppresses the establishment of Campylobacter bacteria. Has obtained knowledge that it works effectively, and has completed the present invention.

  That is, the composition for countermeasure against Campylobacter of the present invention contains β-1,4-mannobiose and is used for countermeasure against Campylobacter.

  In this composition, β-1,4-mannobiose may be derived from a mannan-containing natural product. Further, the mannan-containing natural product may be derived from a lionaceae plant. Further, β-1,4-mannobiose may be produced by allowing a mannan-containing natural product to act on a mannan-degrading enzyme. Moreover, while further containing mannose, more β-1,4-mannobiose may be contained than mannose.

  The additive for feed for the countermeasure against Campylobacter according to the present invention contains the above-mentioned composition against Campylobacter.

  The Campylobacter countermeasure feed of the present invention contains the above-mentioned Campylobacter countermeasure composition or feed additive.

  The method for using the β-1,4-mannobiose-containing composition of the present invention includes the step of feeding the β-1,4-mannobiose-containing composition to livestock infected with Campylobacter bacteria.

In addition, another method of using the β-1,4-mannobiose-containing composition of the present invention includes a step of feeding the β-1,4-mannobiose-containing composition before transporting livestock.
In these methods of use, the livestock may be a broiler.

  According to the present invention, the β-1,4-mannobiose campylobacter recognition activity and the β-1,4-mannobiose are not easily assimilated by the bacteria in the intestines of the livestock. Can be effectively suppressed. Therefore, here, it is possible to cope with the effects that occur in livestock due to Campylobacter bacteria by safe and simple means.

  The campylobacter composition of the present invention contains β-1,4-mannobiose (hereinafter, also simply referred to as mannobiose), and is used as a campylobacter countermeasure.

  In the present invention, β-1,4-mannobiose is formed by bonding two mannose molecules to β-1,4-glycoside. The β-1,4-mannobiose used in the present invention can be obtained, for example, by a method of synthesizing from mannose or a method of decomposing β-1,4-mannan (hereinafter also simply referred to as mannan).

  The method for decomposing β-1,4-mannan is more preferable in terms of the raw material resources and reaction efficiency, and β-1,4-mannobiose can be obtained more easily. In this method, for example, a mannan-degrading enzyme is allowed to act on mannan-containing natural products such as palm kernel meal, copra meal, guar gum, locust bean gum and the like, which are rich in mannan, or mannan extracted from these natural products. , 4-Mannobiose can be obtained.

  The mannan degrading enzyme used in this method may be any mannanase, mannosidase, hemicellulase, etc., as long as it has an activity of degrading mannan to produce mannobiose, derived from Aspergillus niger, Commercially available products (for example, hemicellulase GM “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.), Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd.), cellulosin GM5 (Hankyu Bioindustry Co., Ltd.) and the like can be preferably used. In addition to these, even those commercially available as xylanase and cellulase, those having the hydrolysis activity can be used, for example, cellulase Y-NC (manufactured by Yakult Pharmaceutical Co., Ltd.) can be used. (Exo type) low activity, mannanase (endo type) activity Hemicellulase GM “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.) and Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd.) are preferred because they can suppress the production of mannose and can produce a large amount of mannobiose.

  Furthermore, in this method, the mannan degrading enzyme is allowed to act on mannan-containing natural products or mannan extracted therefrom as an enzyme solution dissolved or dispersed in water. In order to perform an efficient reaction in the case of using a mannan-containing natural product, it is important to adjust moisture in a reaction system including the mannan-containing natural product, a mannan-degrading enzyme, and water. The amount of water added to adjust the water content is preferably 50 to 10,000 parts by weight, more preferably 50 to 1500 parts by weight, with respect to 100 parts by weight of mannan. By setting the amount of water to be in such a range, the mannan fibers can be sufficiently swollen in the presence of sufficient moisture, and the enzyme solution can be easily contacted. However, an excessive amount of water not only decreases the reaction efficiency as a result of diluting the enzyme concentration, but also increases the drying cost in the drying step when drying. Therefore, considering the above, it is appropriate to add 50 to 500 parts by weight of water to 100 parts by weight of mannans.

  The amount of the enzyme and the reaction time are at least 10% by weight of the mannobiose produced with respect to the mannan before decomposition, and preferably the mannobiose produced by hydrolysis is about 10 to 80% by weight with respect to the mannan before decomposition. If it is a thing, it will not be specifically limited, Under such conditions, a wet enzyme processed material can be obtained. However, since an enzyme having a high mannanase (endo type) activity usually also has a mannosidase (exo type) activity, if the enzyme reaction time is too long, mannobiose is decomposed and the amount of mannose increases. Therefore, the reaction time is preferably not longer than necessary. These enzyme reaction conditions are appropriately set so that the amount of mannobiose produced is as large as possible. In this case, it is preferable to set so that β-1,4-mannobiose is contained more than mannose. For example, the ratio of mannose to β-1,4-mannobiose is more preferably 60% by weight or less, and 20 It is particularly preferred that it is not more than wt%.

  As described above, for example, when palm kernel meal (mannan content is approximately 36%) is used as a raw material and reacted for 3 to 36 hours, the amount of mannobiose depends on the type, amount, and time of the enzyme used. However, it can be produced up to about 6 to 17 parts by weight per 100 parts by weight of the raw material.

  In order to suppress colonization of Campylobacter bacteria in the intestines of livestock, the enzyme-treated product containing mannobiose thus produced or the water extract of the enzyme-treated product (β-1,4-mannobiose-containing composition) It can be used as an additive for feed by blending additive components, or it can be blended and used as it is in feed. For example, when the composition is a water extract, it is used by adding it to beverages. be able to. The water extract contains mannose and mannooligosaccharides in addition to mannobiose, but it is not necessary to extract and purify only mannobiose, and it is preferable that these are contained.

  The additive for feed for the countermeasure against Campylobacter according to the present invention may be one in which an additive component such as a preservative is blended with the β-1,4-mannobiose-containing composition, and is used as a countermeasure against Campylobacter. It is what When such a feed additive is used, since a large amount of water is contained, there is a risk of generation of fungi and fungi. Therefore, the water content is 10% or less by a method such as fluidized bed drying or vacuum drying. It is preferable to dry so that it becomes.

  The Campylobacter countermeasure feed of the present invention is a feed comprising the above composition or feed additive, and is used as a Campylobacter countermeasure. For example, β-1,4-mannobiose-containing copra meal may be used as it is as the feed or may be blended with other feeds. When adding β-1,4-mannobiose-containing copra meal to the mixed feed, the amount of the β-1,4-mannobiose content in the mixed feed is 0.001 to 0.6% by weight, usually in the mixed feed. On the other hand, it is desirable to add β-1,4-mannobiose-containing copra meal to 0.01 to 2% by weight, preferably 0.05 to 1% by weight. By feeding such a feed to livestock in a solid or liquid state, it is possible to suppress the establishment of Campylobacter bacteria in the animal body and efficiently discharge Campylobacter bacteria outside the body. The livestock includes poultry such as broilers (edible young chickens).

  In the present invention, the Campylobacter composition, the feed additive, and the feed may be used in a situation where at least Campylobacter bacteria may be present in the intestines of livestock, such as Salmonella It may be used in the situation where other bacteria coexist.

Next, how to use the β-1,4-mannobiose-containing composition will be described.
The method of using the β-1,4-mannobiose-containing composition of the present invention includes the step of feeding the β-1,4-mannobiose-containing composition to livestock infected with Campylobacter bacteria, Livestock infected with Pyrobacter can be treated. Here, feeding the β-1,4-mannobiose-containing composition is not limited to feeding the β-1,4-mannobiose composition as it is, and the β-1,4-mannobiose composition was blended. Feeding as a feed additive or feed is also included.

  In addition to or in combination with this method, it is also possible to include a step of feeding the β-1,4-mannobiose-containing composition before transporting livestock. In this case, before transporting livestock, it is possible to clean up livestock contaminated with Campylobacter bacteria and prevent infection of livestock with Campylobacter bacteria. Here, the term “transport” includes shipment to the market, movement to a different breeding place, and the like. Moreover, when it says before conveyance, it may be just before several hours ago, or may be before a comparatively long period like several days ago.

  EXAMPLES Hereinafter, although the Example of this invention is shown and this invention is demonstrated in detail, this invention is not limited to a following example. “Part” and “%” in the examples represent “part by weight” and “% by weight”, respectively, unless otherwise specified.

  150 parts of an enzyme solution obtained by dissolving 0.25 parts of the enzyme hemicellulase GM “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.) in 100 parts of copra meal (30% mannan content, 4.2% moisture) at 60 ° C. for 12 hours. After the action, it was dried to a moisture content of 9.3% using a fluidized bed drying apparatus (manufactured by Okawara Seisakusho) to obtain 106 parts of dry powder. When the mannose content and β-1,4-mannobiose content of this dry powder were measured by ion exchange chromatography, 1.36 parts of mannose and 13.35 parts of β-1,4-mannobiose (in mannan) 44.5%).

Comparative Example 1

  150 parts of an enzyme solution in which 0.3 part of the enzyme cellulosin GM5 (manufactured by HI Corporation) was dissolved in 100 parts of palm kernel meal (mannan content 38%, moisture 4.5%) was allowed to act at 60 ° C. for 72 hours. Then, it dried with the fluidized bed drying apparatus to 7.8% of moisture, and obtained 100 parts of dry powders. When the mannose content and β-1,4-mannobiose content of this dry powder were measured, mannose was 11.52 parts and β-1,4-mannobiose was 2.57 parts (7.14% with respect to mannan). ) Was generated.

(Animal test 1)
Thirty commercially available broiler primary messines were divided into an additive-free control group, an example group, and a comparative example group, and were bred with a commercial broiler mixed feed, with an unrestricted feeding and free drinking water. At 7 days of age, Campylobacter jejuni (CJ), 1 x 10 ⁹ CFU / feather was orally inoculated. At 30 days of age, one bird was dissected for each section, and the number of CJ in the cecum contents was measured. And confirmed the infection. Furthermore, from the 35th day of age, the feed was switched to the following test feed, which differs for each section.

  The additive-free mixed feed was fed as it was to 10 birds in the non-control group, and 0.1% of the dry powder obtained in Example 1 was added to the mixed feed in 10 birds in the example zone. The 10 feeds in the comparative example section were fed a feed supplemented with 0.1% of the dry powder obtained in Comparative Example 1. Thereafter, at 36 days, 38 days, and 41 days of age, two birds were necropsied in each group, and the number of CJs (logarithm) in the contents of the cecum was examined.

  The result is shown in FIG. As shown in the figure, the number of bacteria decreased from 36 days of age immediately after switching to the test feed only in the example section, and the number of bacteria decreased significantly at 38 days and 41 days of age. It can be seen that it has an excellent effect on fixing suppression.

  In addition, Table 1 shows changes in the number of CJs (logCFU / g) in the cecal contents of each female chick.

(Animal test 2)
Eighty commercial broiler brooder messels were brought in and reared on a commercial feed for γ-ray sterilized broilers and well water by continuous feeding. At 0 days of age, nalidixic acid-resistant Salmonella enteritidis (hereinafter referred to as SE) 6.7 × 10 ⁶ CFU / feather and Campylobacter (CJ) 1.1 × 10 ⁷ CFU / feather were simultaneously orally inoculated. Sixty-five birds that were 7 days old and approximated body weight were selected, and SE 9.2 × 10 ⁸ CFU / feather and CJ1.0 × 10 ⁹ CFU / feather were simultaneously orally inoculated again. Furthermore, 50 birds were selected at the age of 20 days, and 12 birds were assigned to 4 groups, and 2 groups were assigned to 2 test areas. At the age of 21 days, SE8.6 × 10 ⁸ CFU / feather and CJ1.0 × 10 ⁹ CFU / feather were orally inoculated simultaneously. The remaining 2 birds not belonging to the test area were collected at 27 days of age, and SE and CJ of the cecum contents were quantified to obtain the initial contamination concentration of each group. And one of the two test groups is a positive control (non-addition control group) and continues to feed the commercial feed until the end of the 34-day-old test, and the other test group (Example group) is 27 days old. Until the age of 34 days, 0.1% of the dry powder (enzyme-treated copra meal, β-mannobiose type) obtained in Example 1 was added to the commercial feed and fed. Three days (6 birds in each group) were randomly selected at 1 day (28 days old), 3 days (30 days old), 7 days (34 days old) after starting feeding the test samples, and the contents of the cecum were determined. The sample was collected and SE and CJ were quantified.
The result is shown in FIG. In the example group, SE and CJ are both lower than the non-added control group from the day after the start of feeding, and it can be seen that the product of the present invention works effectively even when two food poisoning causative bacteria are simultaneously infected.
Table 2 shows the transition of the number of bacteria (logCFU / g) in the cecal contents of each chick.

The graph which shows the CJ number (logCFU / g) in the cecum of the female chick in the predetermined day age in Example 2. FIG. The graph which shows the number of each bacteria (logCFU / g) in the caecum of the female chick in the predetermined age in Example 3. FIG.

Claims (10)

  A composition for preventing Campylobacter, comprising β-1,4-mannobiose.   The composition for campylobacter countermeasures according to claim 1, wherein β-1,4-mannobiose is derived from a mannan-containing natural product.   The composition for countermeasure against Campylobacter according to claim 2, wherein the mannan-containing natural product is derived from a lion family plant.   The composition for preventing Campylobacter according to claim 1, wherein β-1,4-mannobiose is produced by allowing a mannan-containing enzyme to act on a mannan-containing natural product.   The composition for a countermeasure against Campylobacter according to claim 1, further comprising mannose and containing more β-1,4-mannobiose than mannose.   The additive for feed for a Campylobacter countermeasure containing the composition for a Campylobacter countermeasure of any one of Claim 1 to 5.   A Campylobacter-control feed comprising the Campylobacter-control composition according to any one of Claims 1 to 5 or the Campylobacter-control feed additive according to Claim 6.   A method of using a β-1,4-mannobiose-containing composition comprising the step of feeding a β-1,4-mannobiose-containing composition to livestock infected with Campylobacter bacteria.   A method of using a β-1,4-mannobiose-containing composition comprising the step of feeding the β-1,4-mannobiose-containing composition before delivering livestock. The method for using the β-1,4-mannobiose-containing composition according to claim 8 or 9, wherein the livestock is a broiler.

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