KR101945934B1 - Feed additive composition for prevention or treatment of Salmonella diarrhea comprising organic mineral complex - Google Patents

Feed additive composition for prevention or treatment of Salmonella diarrhea comprising organic mineral complex Download PDF

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KR101945934B1
KR101945934B1 KR1020160139419A KR20160139419A KR101945934B1 KR 101945934 B1 KR101945934 B1 KR 101945934B1 KR 1020160139419 A KR1020160139419 A KR 1020160139419A KR 20160139419 A KR20160139419 A KR 20160139419A KR 101945934 B1 KR101945934 B1 KR 101945934B1
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노혜지
허수정
고홍범
김희경
조향현
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(주)비티엔
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/153—Nucleic acids; Hydrolysis products or derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/06—Aluminium, calcium or magnesium; Compounds thereof, e.g. clay
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/30—Zinc; Compounds thereof
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    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract

본 발명은 유기태화 철 및 퓨린염기 함유 핵산 단량체의 복합체를 유효성분으로 하는 살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물을 제공하기 위한 것으로, 상기 사료 첨가제 조성물은 항생제를 대체하여 살모넬라성 설사 예방 또는 치료용으로 사용될 수 있다.The present invention provides a feed additive composition containing an organic minerals complex for preventing or treating Salmonella diarrhea comprising a complex of a nucleic acid monomer containing an organotissive iron and a purine base as an active ingredient, wherein the feed additive composition is a salmonella Can be used for preventing or treating diarrhea.

Description

살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물 {Feed additive composition for prevention or treatment of Salmonella diarrhea comprising organic mineral complex}Technical Field [0001] The present invention relates to a feed additive composition for preventing or treating diarrhea caused by salmonella,

본 발명은 살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물에 관한 것이다. 특히 철분이 퓨린염기 함유 핵산 단량체에 킬레이트 결합된 유기태화 미네랄 복합체를 유효성분으로 하는 사료 첨가제 조성물에 관한 것이다.The present invention relates to a feed additive composition containing organotinized mineral complex for preventing or treating Salmonella diarrhea. The present invention relates to a feed additive composition comprising an organic minerals complex in which iron is chelated with a purine base-containing nucleic acid monomer as an active ingredient.

동물에서 항생제는 세균성 감염증의 예방 및 치료에 사용될 뿐만 아니라 가축의 발육촉진과 생산성 향상을 목적으로 사용되어 왔으나 항생제의 오남용으로 인한 많은 종류의 세균들이 내성을 갖게 되어 보다 높은 농도의 사용과 더불어 새로운 항생제의 사용이 요구되고 있다. 이러한 무분별한 항생제의 사용으로 인해 항생제에 대한 내성균의 증가와 생산물에 대한 잔류문제가 대두되면서 전세계적으로 사람의 건강과 안전성 문제로 인하여 점차 항생제의 사용을 규제하고 있으며 그 규제 정도도 점점 엄격해지고 있는 실정이다. 안전한 먹거리 생산에 대한 소비자들의 관심이 높아지고 축산분야의 항생제 사용의 문제점이 대두됨에 따라 천연 유래 항생제에 관한 관심이 증대되고 있다.Antibiotics in animals have been used not only for the prevention and treatment of bacterial infections but also for the purpose of promoting the development of livestock and improving productivity. However, since many types of bacteria are resistant to abuse of antibiotics, Is required. The use of these indiscriminate antibiotics has led to an increase in resistant bacteria to antibiotics and a residual problem with products, and the use of antibiotics is increasingly regulated due to human health and safety problems around the world. to be. Consumers' interest in safe food production has increased and interest in antibiotics derived from natural sources has increased as the problem of using antibiotics in the livestock sector has emerged.

한국공개특허 제10-2016-0043480호는 유기태화 미네랄 및 핵산 복합체의 제조방법에 관한 것으로, 상기 방법으로 제조된 유기태화 미네랄 및 핵산 복합체는 무기태 미네랄을 대체하면서, 무기태 미네랄에 비해 생체이용율이 뛰어나므로 소량 사용만으로도 충분한 미네랄 공급 효과를 달성하면서 과량의 무기태 미네랄 사용으로 인한 변색, 이미, 이취의 발생을 방지할 수 있다고 기재하고 있으나, 단순히 미네랄의 공급 효과 외에 항생제 대체 효과에 대해서는 어떠한 기재도 없었다.Korean Patent Laid-Open No. 10-2016-0043480 relates to a method for producing an organotinized mineral and a nucleic acid complex, wherein the organotinized mineral and nucleic acid complexes prepared by the above method are used in place of inorganic minerals, It is described that it is possible to prevent the discoloration, already, and the occurrence of odor due to the use of excessive inorganic minerals while achieving a sufficient mineral supply effect only by a small amount of use. However, in addition to the effect of supplying minerals, There was no.

본 발명자들은 상기 한국공개특허 제10-2016-0043480호에서 미네랄 공급용으로 제시되었던 유기태화 미네랄 및 핵산 복합체 중 일부가 살모넬라성 설사 예방 또는 치료용으로 사용될 수 있음을 처음으로 확인하여 본 발명을 완성하였다.The present inventors have confirmed for the first time that some of the organic minerals and nucleic acid complexes proposed for mineral supply in Korean Patent Laid-open No. 10-2016-0043480 can be used for the prevention or treatment of salmonellosis, Respectively.

본 발명의 목적은 항생제를 대체하여 살모넬라성 설사 예방 또는 치료용으로 사용될 수 있는 사료 첨가제 조성물을 제공하기 위한 것이다.It is an object of the present invention to provide a feed additive composition which can be used for preventing or treating Salmonella diarrhea by replacing antibiotics.

본 발명은 유기태화 철 및 퓨린염기 함유 핵산 단량체의 복합체를 유효성분으로 하는 살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물을 제공한다.The present invention provides a feed additive composition containing an organic minerals complex for preventing or treating Salmonella diarrhea comprising a complex of a nucleic acid monomer containing an organotissive iron and a purine base as an active ingredient.

상기 유기태화 철 및 퓨린염기 함유 핵산 단량체의 복합체는 칼슘, 아연, 마그네슘, 칼륨, 구리, 셀렌, 크롬, 몰리브덴 및 요오드 중에서 선택되는 어느 하나 또는 둘 이상의 미네랄이 추가로 킬레이트된 유기태화 미네랄 및 퓨린염기 함유 핵산 단량체의 복합체일 수 있다.The complex of the organotissive iron and the purine base-containing nucleic acid monomer may be any one or two or more of minerals selected from calcium, zinc, magnesium, potassium, copper, selenium, chromium, molybdenum and iodine, Containing nucleic acid monomer.

상기 유기태화 철 및 퓨린염기 함유 핵산 단량체는 퓨린염기 함유 핵산 단량체 또는 이의 염 분말, 및 용해엔탈피가 음성인 수용성 철 분말을 혼합한 후, 상기 혼합물에 물을 혼합하여 반응시키는 단계; 또는 퓨린염기 함유 핵산 단량체 또는 이의 염 분말과 물을 혼합하여 용해시킨 후, 상기 수용액에 용해엔탈피가 음성인 수용성 철 분말을 혼합하여 반응시키는 단계;를 포함하여 제조될 수 있다.Wherein the nucleic acid monomer containing the iron oxyhydrogen and the purine base is prepared by mixing a nucleic acid monomer containing a purine base or a salt thereof and a water soluble iron powder having a negative enthalpy of dissolution and then mixing and reacting the mixture with water; Or a purine base-containing nucleic acid monomer or a salt thereof and water, and mixing the aqueous solution with a water-soluble iron powder having a negative enthalpy of dissolution.

상기 칼슘, 아연, 마그네슘, 칼륨, 구리, 셀렌, 크롬, 몰리브덴 및 요오드 중에서 선택되는 어느 하나 또는 둘 이상의 미네랄이 추가로 킬레이트된 유기태화 미네랄 및 퓨린염기 함유 핵산 단량체의 복합체를 제조할 경우에는, 상기 유기태화 철 및 퓨린염기 함유 핵산 단량체는 퓨린염기 함유 핵산 단량체 또는 이의 염 분말, 및 용해엔탈피가 음성인 수용성 철 분말을 혼합하는 과정에서 상기 수용성 철 분말 이외에 추가로 킬레이트시킬 용해엔탈피가 음성인 수용성 미네랄 분말을 혼합할 수 있다.When a complex of an organic minerals and purine base-containing nucleic acid monomers further chelated with any one or two or more minerals selected from calcium, zinc, magnesium, potassium, copper, selenium, chromium, molybdenum and iodine is produced, The nucleic acid monomer containing organotin ferrous and purine bases may be further mixed with water-soluble minerals which are chelated in addition to the water-soluble iron powder in the course of mixing the purine base-containing nucleic acid monomer or its salt powder and the water- The powder can be mixed.

상기 용해엔탈피가 음성인 수용성 미네랄 분말은 복수의 서로 다른 미네랄을 순차적으로 또는 동시에 퓨린염기 함유 핵산 단량체 또는 이의 염 분말과 혼합할 수 있다. 상기 용해엔탈피가 음성인 수용성 미네랄 분말은 통상의 기술자에게 널리 알려져 있는 것을 사용할 수 있다.The water soluble mineral powder having a negative dissolution enthalpy can be mixed with a plurality of different minerals sequentially or simultaneously with a purine base-containing nucleic acid monomer or a salt thereof. The water-soluble mineral powder in which the dissolution enthalpy is negative may be those well-known to those skilled in the art.

상기 용해엔탈피가 음성인 수용성 미네랄의 경우 물과 혼합되었을 때, 특히 소량의 물과 혼합되었을 때 이온화되지 않고 발열 반응을 일으키면서 유기태화에 필요한 기질이 존재할 경우 킬레이트 결합을 형성한다.In the case of water-soluble minerals having negative enthalpy of dissolution, they form chelate bonds when they are mixed with water, especially when they are mixed with a small amount of water, when they are not ionized and generate an exothermic reaction while a substrate necessary for organicization is present.

상기 퓨린염기 함유 핵산 단량체는 퓨린환의 6번 위치에 수산기(-OH)를 포함하는 퓨린염기가 용해엔탈피가 음성인 수용성 미네랄 분말과 결합하여 불용성 침전물을 형성하고, 유기태화 미네랄 및 핵산 복합체의 생산 수율이 높다는 점에서 바람직하다. 예를 들어 상기 퓨린염기 함유 핵산 단량체는 이노신산, 구아닐산 및 크산틸산 중에서 선택되는 어느 하나 이상의 핵산 단량체일 수 있고, 바람직하게는 이노신산 또는 구아닐산이다. 가장 바람직하게는 이노신산의 나트륨염 또는 구아닌산의 나트륨염이다.The purine base-containing nucleic acid monomer has a purine base containing a hydroxyl group (-OH) at the 6-position of the purine ring, forming an insoluble precipitate by binding to a water-soluble mineral powder having a negative enthalpy of dissolution, and the production yield of the organotinized mineral and nucleic acid complex Is high. For example, the purine base-containing nucleic acid monomer may be any one or more nucleic acid monomers selected from inosine acid, guanylic acid and xanthic acid, preferably inosinic acid or guanylic acid. Most preferably the sodium salt of inosinic acid or the sodium salt of guanine acid.

상기 물은 식품, 의약, 사료 또는 비료나 그 첨가제에 사용되는 물이면 특별히 제한할 필요가 없으나, 바람직하게는 다른 금속염의 영향을 적게하기 위해 탈이온수를 이용하는 것이 바람직하다.The water is not particularly limited as long as the water is used for foods, medicines, feeds or fertilizers or additives thereof, but it is preferable to use deionized water in order to reduce the influence of other metal salts.

상기 퓨린염기 함유 핵산 단량체 또는 이의 염 분말, 용해엔탈피가 음성인 수용성 철 및/또는 철 이외의 추가 미네랄 분말, 및 물의 혼합 순서에 관계없이, 상기 퓨린염기 함유 핵산 단량체 또는 이의 염 분말, 및 용해엔탈피가 음성인 수용성 철 및/또는 철 이외의 추가 미네랄 분말의 혼합비율은 1:5 내지 95:1 중량비, 바람직하게는 1:2 내지 10:1 중량비, 더욱 바람직하게는 1:1 내지 5:1 중량비일 수 있고, 유기태화 미네랄 및 핵산 복합체의 킬레이트화된 미네랄의 목표 함량에 따라 조절할 수 있다.Irrespective of the mixing order of the purine base-containing nucleic acid monomer or its salt powder, the water-soluble iron and / or iron other than the iron having a negative enthalpy of dissolution, and the water, and the salt powder and the dissolution enthalpy The mixing ratio of the water-soluble iron and / or the additional mineral powder other than iron is 1: 5 to 95: 1 by weight, preferably 1: 2 to 10: 1 by weight, more preferably 1: 1 to 5: And can be adjusted according to the target content of the chelated minerals of the organo-minerals and nucleic acid complexes.

상기 퓨린염기 함유 핵산 단량체 또는 이의 염 분말, 용해엔탈피가 음성인 수용성 철 및/또는 철 이외의 추가 미네랄 분말, 및 물의 혼합 순서에 관계없이, 상기 용해엔탈피가 음성인 수용성 철 및/또는 철 이외의 추가 미네랄 분말과 물의 혼합비율은 1:50 내지 2:1 중량비, 바람직하게는 1:10 내지 1:1 중량비일 수 있다. 상기 물의 함량은 반응을 위해 적절히 조절하면 되지만 너무 많으면 발열반응을 통한 미네랄의 킬레이트화에 바람직하지 못하고, 너무 적으면 미네랄 및 핵산의 혼합물을 균일하게 수화하는데 한계가 있을 수 있다.Irrespective of the mixing order of the purine base-containing nucleic acid monomer or salt powder thereof, the water-soluble iron and / or the additional mineral powder other than iron having a negative enthalpy of dissolution, and water, The mixing ratio of the additional mineral powder and water may be 1:50 to 2: 1 by weight, preferably 1:10 to 1: 1 by weight. The content of water may be appropriately adjusted for the reaction, but if it is too much, it is not preferable for chelating the mineral through the exothermic reaction, and if it is too small, there may be a limit to uniformly hydrating the mixture of mineral and nucleic acid.

상기 물을 첨가하여 반응시키는 과정은 10 내지 100 ℃에서 30 분 내지 5일 동안 수행될 수 있다. 특별히 반응시키는 과정에서 별도의 가열을 하더라도 생산 수율의 증가나 킬레이트된 미네랄 함량의 증가에 유리하다고 할 수 없고, 반응시간은 특별히 한정할 필요없이 반응이 충분히 이루어질 정도의 시간, 예를 들어 1 시간 이상, 2 시간 이상, 4 시간 이상, 6 시간 이상, 12 시간 이상으로 조절할 수 있고, 최대 반응시간도 3일, 2일, 1일로 조절할 수 있다.The process of adding and reacting the water may be performed at 10 to 100 ° C for 30 minutes to 5 days. Even if the reaction is carried out in a special reaction, it is not advantageous to increase the production yield or to increase the chelated mineral content. The reaction time is not particularly limited, and the reaction time can be sufficiently long, for example, 1 hour or more , More than 2 hours, more than 4 hours, more than 6 hours, more than 12 hours, and the maximum reaction time can be adjusted to 3 days, 2 days, 1 day.

상기 수용성 미네랄 분말로 염화물을 사용하는 경우 미네랄이 킬레이트화되는 양이 증가함에 따라 염소 이온의 함량이 증가한다. 이러한 증가된 염소 이온은 물로 세척하는 공정으로 간단히 제거될 수 있다. 예를 들어 염화칼슘을 용해하기 위해 사용된 물의 부피의 0.5 ~ 10 배의 물, 바람직하게는 1 ~ 2 배의 물을 사용하여 1 ~ 5회 반복 세척을 통해 염소 이온의 농도를 무해한 수준으로 낮출 수 있다. 상기 세척은 적절한 양의 물을 첨가한 후 원심분리하여 상등액을 제거하는 방식, 예를 들어 500 ~ 50,000 rpm에서 1 ~ 60 분 원심분리할 수 있고, 이러한 원심분리과정은 온도 조건 설정없이 실온에서 수행될 수 있으나, 25 ℃ 이하, 바람직하게는 15 ℃ 이하로 온도를 일정하게 설정하여 수행될 수 있다.When chlorides are used as the water-soluble mineral powders, the content of chlorine ions increases as the amount of minerals chelated increases. These increased chlorine ions can be simply removed by washing with water. For example, the concentration of chlorine ions may be lowered to a harmless level by repeating 1-5 repeated times using 0.5 to 10 times of water, preferably 1 to 2 times, of the volume of water used to dissolve calcium chloride have. The washing may be performed by adding water in an appropriate amount and centrifuging to remove the supernatant. For example, centrifugation may be performed at 500 to 50,000 rpm for 1 to 60 minutes. The centrifugation may be performed at room temperature But may be performed at a constant temperature of 25 占 폚 or lower, preferably 15 占 폚 or lower.

상기 본 발명의 유기태화 미네랄 및 핵산 복합체는 열풍건조, 분무건조, 동결건조 등 적절한 방법을 통해 건조시켜 분말 형태로 이용하는 것이 사용에 편리하다.The organic minerals and nucleic acid complexes of the present invention may be conveniently used in the form of a powder by drying through a suitable method such as hot air drying, spray drying or freeze drying.

상기 유기태화 미네랄 및 퓨린염기 함유 핵산 단량체의 복합체는 유기태화 칼슘과 철 및 퓨린염기 함유 핵산 단량체의 복합체이고, 상기 칼슘과 철분의 중량비는 5:1 내지 1:5인 것이 바람직하다.The complex of the organotinized mineral and the purine base-containing nucleic acid monomer is a complex of organotaxated calcium and an iron and purine base-containing nucleic acid monomer, and the weight ratio of the calcium to the iron is preferably 5: 1 to 1: 5.

본 발명의 유기태화 철 및 퓨린염기 함유 핵산 단량체의 복합체는 유기태화 기질로 유단백질이나 키토산을 사용한 유기태화 철 및 유단백질 복합체, 유기태화 철 및 키토산 복합체 및 다른 미네랄을 유기태화시킨 유기태화 미네랄 및 퓨린염기 함유 핵산 단량체의 복합체와 비교했을 때 살모넬라 접종 후 사료 섭취량 및 산란율을 유지시키고, 파란율을 현저히 감소시키며, 체중이 덜 감소되도록 하며, 비장종대를 억제하고, 바닥 분변 내 살모넬라 균수에서 확인되듯이 장내 살모넬라 균수를 감소시키고, T세포를 활성화시키고, IL-2 및 인터페론-γ의 분비를 현저히 증가시키므로, 항생제를 대체하여 살모넬라성 설사 예방 또는 치료용으로 사용될 수 있다.The complex of the organotinized iron and purine base-containing nucleic acid monomers of the present invention can be used as an organizing substrate for organotinized minerals and purine bases, which are organicized iron and milk protein complexes using milk proteins or chitosan, organotinized iron and chitosan complexes and other minerals Compared to the complex of nucleic acid monomers containing Salmonella, it is possible to maintain the feed intake and egg production rate after salmonella inoculation, significantly reduce the blue ratio, reduce the body weight to a lesser extent, suppress splenic enlargement, Can be used to prevent or treat Salmonella diarrhea by replacing antibiotics because it reduces Salmonella bacteria, activates T cells, and significantly increases the secretion of IL-2 and interferon-gamma.

이하 본 발명을 실시예, 실험예 및 제조예에 의해 상세히 설명한다. 단, 하기 실시예, 실험예 및 제조예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예, 실험예 및 제조예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples. However, the following Examples, Experimental Examples and Preparation Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples, Experimental Examples and Production Examples.

제조예: 유기태화 미네랄 복합체의 제조Preparation Example: Preparation of Organo-mineralized Mineral Complex

1) 유기태화 철 및 유단백질 복합체(Fe-WPS, 비교예 1)의 제조1) Preparation of an organotin-iron and milk protein complex (Fe-WPS, Comparative Example 1)

치즈 제조 후 생성되는 유청을 농축 후 건조하여 제조된 유청분말(매일유업, 이하 'WPS'라고도 함)을 시료로 사용하였다. 상기 유청분말의 단백질 함량은 14 중량%, 지방 함량은 7 중량%, 회분 함량은 0.5 중량%, 나머지는 탄수화물이었고, 상기 유청분말은 5 분 이상의 가열 또는 살균 처리를 행하지 않은 것이었다.Whey powder (daily dairy product, hereinafter also referred to as "WPS") prepared by concentrating and drying whey produced after cheese production was used as a sample. The protein content of the whey powder was 14% by weight, the fat content was 7% by weight, the ash content was 0.5% by weight, the remainder was carbohydrate, and the whey powder was not heated or sterilized for 5 minutes or more.

상기 유청분말 10 중량부에 황산제1철[FeSO4·7H2O(Yakuri Pure Chemical)] 분말 5 중량부를 먼저 분말 혼합하고, 상기 혼합된 분말에 정제수 20 중량부를 혼합하여 상온에서 500 rpm으로 교반하면서 24 시간 반응시켰다. 상기 24 시간 동안 반응시킨 유기태화 반응물을 25 ℃에서 3,000rpm으로 20분 원심분리하여 침전물을 얻고, 상기 침전물 중량의 2 배 중량의 정제수를 혼합 및 교반한 후 상기 조건과 동일하게 원심분리하는 세척과정을 5회 반복하여 미반응 유단백질 및 미네랄을 제거한 후, 동결건조하여 유기태화 철 및 유단백질 복합체(Fe-WPS, 비교예 1)를 제조하였다. 5 parts by weight of ferrous sulfate [FeSO 4 .7H 2 O (Yakuri Pure Chemical)] powder was previously mixed with 10 parts by weight of the above whey powder, 20 parts by weight of purified water was mixed with the mixed powder and stirred at 500 rpm at room temperature While reacting for 24 hours. The organic reaction product which has been reacted for 24 hours is centrifuged at 3,000 rpm for 20 minutes at 25 DEG C to obtain a precipitate. The purified water having a weight twice the weight of the precipitate is mixed and stirred, and centrifuged Was repeated five times to remove unreacted milk proteins and minerals, followed by lyophilization to prepare an iron oxide and protein complex (Fe-WPS, Comparative Example 1).

2) 유기태화 철 및 키토산 복합체(Fe-OCHT, 비교예 2)의 제조2) Preparation of organotinized iron and chitosan complex (Fe-OCHT, Comparative Example 2)

키토산[금호화성(DAC 90% 이상, 분자량 10,000이하, 한국)]을 구입하여 (주)한일바이오메드(전남, 한국)에서 키토산올리고당(chitosan oligosaccharide, 이하, OCHT)으로 제조한 후, 키토산올리고당을 기질로 하는 철분을 킬레이팅 시켜 제조한 유기태화 철 및 키토산 복합체(Fe-OCHT, 비교예 2, 상품명 : EPECTROL-3)를 제조하였다.Chitosan oligosaccharide (hereinafter, referred to as OCHT) was purchased from Hanil Biomed (Korea, Jeonnam, Korea), and chitosan oligosaccharide was synthesized from chitosan oligosaccharide (Fe-OCHT, Comparative Example 2, trade name: EPECTROL-3) prepared by chelating iron as a substrate were prepared.

3) 유기태화 철 및 구아닐산 복합체(Fe-GHAC, 실시예 1)의 제조3) Preparation of organotinized iron and guanylate complex (Fe-GHAC, Example 1)

5'-구아닐산이나트륨(GMP) 분말 10 중량부에 황산제1철[FeSO4·7H2O(Yakuri Pure Chemical)] 분말 2 중량부를 먼저 분말 혼합하고, 상기 혼합된 분말에 정제수 20 중량부를 혼합하여 상온에서 500 rpm으로 교반하면서 24 시간 반응시켰다. 상기 24 시간 동안 반응시킨 유기태화 반응물을 25 ℃에서 3,000rpm으로 20분 원심분리하여 침전물을 얻고, 상기 침전물 중량의 2 배 중량의 정제수를 혼합 및 교반한 후 상기 조건과 동일하게 원심분리하는 세척과정을 5회 반복하여 미반응 핵산 및 미네랄을 제거한 후, 동결건조하여 유기태화 철 및 구아닐산 복합체(Fe-GHAC, 실시예 1)를 제조하였다. 2 parts by weight of ferrous sulfate [FeSO 4 .7H 2 O (Yakuri Pure Chemical)] powder was firstly powdered and mixed with 10 parts by weight of disodium 5'-disodium guanylate (GMP). 20 parts by weight of purified water And reacted for 24 hours at room temperature with stirring at 500 rpm. The organic reaction product which has been reacted for 24 hours is centrifuged at 3,000 rpm for 20 minutes at 25 DEG C to obtain a precipitate. The purified water having a weight twice the weight of the precipitate is mixed and stirred, and centrifuged Was repeated 5 times to remove unreacted nucleic acids and minerals, followed by lyophilization to prepare an organotized iron and guanylate complex (Fe-GHAC, Example 1).

4) 유기태화 칼슘과 철 및 구아닐산 복합체(CaFe-GHAC, 실시예 2)의 제조4) Preparation of calcium organotaxate, iron and guanylate complex (CaFe-GHAC, Example 2)

5'-구아닐산이나트륨(GMP) 분말 10 중량부에 염화칼슘 분말[CaCl2·2H2O(Junsei, 일본)] 2 중량부 및 황산제1철[FeSO4·7H2O(Yakuri Pure Chemical)] 분말 2 중량부를 먼저 분말 혼합하고, 상기 혼합된 분말에 정제수 20 중량부를 혼합하여 상온에서 500 rpm으로 교반하면서 24 시간 반응시켰다. 상기 24 시간 동안 반응시킨 유기태화 반응물을 25 ℃에서 3,000rpm으로 20분 원심분리하여 침전물을 얻고, 상기 침전물 중량의 2 배 중량의 정제수를 혼합 및 교반한 후 상기 조건과 동일하게 원심분리하는 세척과정을 5회 반복하여 미반응 핵산 및 미네랄을 제거한 후, 동결건조하여 유기태화 칼슘과 철 및 구아닐산 복합체(CaFe-GHAC, 실시예 2)를 제조하였다. 2 parts by weight of calcium chloride powder [CaCl 2 .2H 2 O (Junsei, Japan)] and 1 part by weight of ferrous sulfate [FeSO 4 .7H 2 O (Yakuri Pure Chemical)] were added to 10 parts by weight of disodium 5'- 2 parts by weight of powder were mixed first, 20 parts by weight of purified water was mixed with the mixed powder, and the mixture was reacted for 24 hours while stirring at 500 rpm at room temperature. The organic reaction product which has been reacted for 24 hours is centrifuged at 3,000 rpm for 20 minutes at 25 DEG C to obtain a precipitate. The purified water having a weight twice the weight of the precipitate is mixed and stirred, and centrifuged Was repeated 5 times to remove unreacted nucleic acids and minerals, followed by lyophilization to prepare an organotissive calcium, iron and guanylate complex (CaFe-GHAC, Example 2).

5) 유기태화 칼슘과 철 및 이노신산 복합체(CaFe-IHAC, 실시예 3)의 제조5) Preparation of calcium organotaxate and iron and inosine complex (CaFe-IHAC, Example 3)

5'-이노신산이나트륨(IMP) 10 중량부에 염화칼슘 분말[CaCl2·2H2O(Junsei, 일본)] 2 중량부 및 황산제1철[FeSO4·7H2O(Yakuri Pure Chemical)] 분말 2 중량부를 먼저 분말 혼합하고, 상기 혼합된 분말에 정제수 20 중량부를 혼합하여 상온에서 500 rpm으로 교반하면서 24 시간 반응시켰다. 상기 24 시간 동안 반응시킨 유기태화 반응물을 25 ℃에서 3,000rpm으로 20분 원심분리하여 침전물을 얻고, 상기 침전물 중량의 2 배 중량의 정제수를 혼합 및 교반한 후 상기 조건과 동일하게 원심분리하는 세척과정을 5회 반복하여 미반응 핵산 및 미네랄을 제거한 후, 동결건조하여 유기태화 칼슘과 철 및 이노신산 복합체(CaFe-IHAC, 실시예 3)를 제조하였다. 2 parts by weight of calcium chloride powder [CaCl 2 .2H 2 O (Junsei, Japan)] and 10 parts by weight of ferrous sulfate [FeSO 4 .7H 2 O (Yakuri Pure Chemical)] powder were added to 10 parts by weight of disodium 5'- And 20 parts by weight of purified water was mixed with the mixed powder, and reacted for 24 hours at room temperature with stirring at 500 rpm. The organic reaction product which has been reacted for 24 hours is centrifuged at 3,000 rpm for 20 minutes at 25 DEG C to obtain a precipitate. The purified water having a weight twice the weight of the precipitate is mixed and stirred, and centrifuged Was repeated five times to remove unreacted nucleic acids and minerals, followed by lyophilization to prepare an organotissive calcium and iron and inosine complex (CaFe-IHAC, Example 3).

6) 유기태화 칼슘 및 구아닐산 복합체(Ca-GHAC, 비교예 3)의 제조6) Preparation of organotaxated calcium and guanylate complex (Ca-GHAC, Comparative Example 3)

5'-구아닐산이나트륨(GMP) 분말 10 중량부에 염화칼슘 분말[CaCl2·2H2O(Junsei, 일본)] 4 중량부를 먼저 분말 혼합하고, 상기 혼합된 분말에 정제수 20 중량부를 혼합하여 상온에서 500 rpm으로 교반하면서 24 시간 반응시켰다. 상기 24 시간 동안 반응시킨 유기태화 반응물을 25 ℃에서 3,000rpm으로 20분 원심분리하여 침전물을 얻고, 상기 침전물 중량의 2 배 중량의 정제수를 혼합 및 교반한 후 상기 조건과 동일하게 원심분리하는 세척과정을 5회 반복하여 미반응 핵산 및 미네랄을 제거한 후, 동결건조하여 유기태화 칼슘 및 구아닐산 복합체(Ca-GHAC, 비교예 3)를 제조하였다. 4 parts by weight of calcium chloride powder [CaCl 2 .2H 2 O (Junsei, Japan)] was firstly powdered and mixed with 10 parts by weight of disodium 5'-guanylate (GMP) powder. 20 parts by weight of purified water was mixed with the mixed powder, The reaction was carried out for 24 hours while stirring at 500 rpm. The organic reaction product which has been reacted for 24 hours is centrifuged at 3,000 rpm for 20 minutes at 25 DEG C to obtain a precipitate. The purified water having a weight twice the weight of the precipitate is mixed and stirred, and centrifuged Was repeated 5 times to remove unreacted nucleic acids and minerals, followed by lyophilization to prepare calcium oxalate and guanylate complex (Ca-GHAC, Comparative Example 3).

7) 유기태화 칼슘과 아연 및 구아닐산 복합체(CaZn-GHAC, 비교예 4)의 제조7) Preparation of calcium organotaxate and zinc and guanylate complex (CaZn-GHAC, Comparative Example 4)

5'-구아닐산이나트륨(GMP) 분말 10 중량부에 염화칼슘 분말[CaCl2·2H2O(Junsei, 일본)] 2 중량부 및 황산아연[ZnSO4·7H2O(빅솔, 한국)] 분말 4 중량부를 먼저 분말 혼합하고, 상기 혼합된 분말에 정제수 20 중량부를 혼합하여 상온에서 500 rpm으로 교반하면서 24 시간 반응시켰다. 상기 24 시간 동안 반응시킨 유기태화 반응물을 25 ℃에서 3,000rpm으로 20분 원심분리하여 침전물을 얻고, 상기 침전물 중량의 2 배 중량의 정제수를 혼합 및 교반한 후 상기 조건과 동일하게 원심분리하는 세척과정을 5회 반복하여 미반응 핵산 및 미네랄을 제거한 후, 동결건조하여 유기태화 칼슘과 아연 및 구아닐산 복합체(CaZn-GHAC, 비교예 4)를 제조하였다. 2 parts by weight of calcium chloride powder [CaCl 2 .2H 2 O (Junsei, Japan)] and 10 parts by weight of zinc sulfate [ZnSO 4 .7H 2 O (Bixol, Korea)] were added to 10 parts by weight of disodium 5'-disodium guanylate (GMP) And 20 parts by weight of purified water was mixed with the mixed powder, and the mixture was reacted for 24 hours at room temperature with stirring at 500 rpm. The organic reaction product which has been reacted for 24 hours is centrifuged at 3,000 rpm for 20 minutes at 25 DEG C to obtain a precipitate. The purified water having a weight twice the weight of the precipitate is mixed and stirred, and centrifuged Was repeated five times to remove unreacted nucleic acids and minerals, followed by lyophilization to prepare calcium oxalate and a zinc and guanylate complex (CaZn-GHAC, Comparative Example 4).

실험예 1: 유기태화 미네랄 복합체의 미네랄 분석Experimental Example 1: Mineral analysis of organotinized mineral complex

상기 실시예 1 내지 4 및 비교예 1 내지 3의 유기태화 미네랄 및 핵산 복합체의 IPC 분석을 실시하여 유기태화 칼슘, 철 및 아연 함량을 ppm으로 나타내었다.The organotinized minerals and nucleic acid complexes of Examples 1 to 4 and Comparative Examples 1 to 3 were subjected to IPC analysis to show the contents of calcium, iron and zinc in ppm.

구분division 비교예1Comparative Example 1 비교예2Comparative Example 2 실시예1Example 1 실시예2Example 2 실시예3Example 3 비교예3Comparative Example 3 비교예4Comparative Example 4 약어Abbreviation Fe-WPSFe-WPS Fe-OCHTFe-OCHT Fe-GHACFe-GHAC CaFe-GHACCaFe-GHAC CaFe-IHACCaFe-IHAC Ca-GHACCa-GHAC CaZn-GHACCaZn-GHAC CaCa 1,9901,990 2,9992,999 NDND 51,78051,780 45,38045,380 90,160 90,160 124,400124,400 FeFe 19,120 19,120 359359 16,720 16,720 34,25034,250 27,76027,760 31 31 21 21 ZnZn 77 2,9312,931 44 NDND 99 NDND 31,26031,260

* ND: Not Detected* ND: Not Detected

실험예 2: 산란계에서 유기태화 미네랄 복합체의 살모넬라 공격접종에 따른 효과 확인Experimental Example 2: Effect of Organic-Mineral Complex on Salmonella Inoculation in Laying Hens

1) 사양시험 조건1) Specifications Test conditions

가. 공시동물 및 실험포end. Disclosed animals and laboratory animals

8개월령 된 Hy-line brown 산란계 44마리를 이용하였다. 실험에서 산란계는 실시예 1 내지 4 및 비교예 1 내지 3의 유기태화 미네랄 및 핵산 복합체를 임의 배치하여 이용하였다. 실험동물은 각 실험군별로 분리하여 사육하였으며 물과 사료는 자유 급여하였다. 모든 동물실험 과정은 전남대학교 수의과대학 실험동물사에서 동물실험윤리위원회의 승인을 받아 시행하였다(동물실험승인번호 : CNU IACUC-YB-2015-59).Forty-nine months old Hy-line brown laying hens were used. In the experiment, laying hens were used to arbitrarily arrange the organic minerals and nucleic acid complexes of Examples 1 to 4 and Comparative Examples 1 to 3. Experimental animals were separated for each experimental group and water and feed were fed free. All animal experimental procedures were approved by the Animal Experimental Ethics Committee of the Experimental Animal Husbandry College of Veterinary Medicine, Chonnam National University (CNU IACUC-YB-2015-59).

나. 사료조성, 및 유기태화 미네랄 및 핵산 복합체 혼합량I. Feed composition, and organic minerals and nucleic acid complex mixture

일반사료는 현대사료 주식회사의 협조를 얻어 산란계 산란초기 33호 가루사료에서 미네랄이 제외된 것을 일반사료로 사용하였으며, 미네랄이 제외된 일반사료 1kg에 실시예 1 내지 4 및 비교예 1 내지 3의 유기태화 미네랄 및 핵산 복합체 Fe-WPS는 172mg, Fe-OCHT는 660mg, Fe-GHAC 195 mg, CaFe-GHAC는 97mg, CaFe-IHAC는 97mg, Ca-GHAC는 72mg, 및 CaZn-GHAC는 169mg을 각각 혼합하고, 사료 1톤 당 미네랄 함량이 칼슘은 261.8g, Fe는 183.3g, Zn은 251.43g, Cu는 16.18g 및 Se는 0.7g이 되도록, 무기태 미네랄(탄산칼슘, 황산제1철, 황산아연, 황산동, 소듐셀레나이트, 황산망간 및 요오드화칼슘)을 추가로 첨가하여 미네랄 조성을 동일하게 조정하였다.The general diet was prepared with the help of Hyundai Feed Co., Ltd., And the minerals were excluded from the egg powder 33 of the egg laying period. GHAC, and CaZn-GHAC were mixed at a concentration of 172 mg, 660 mg of Fe-OCHT, 195 mg of Fe-GHAC, 97 mg of CaFe-IHAC, 72 mg of Ca-GHAC and 169 mg of CaZn- Minerals such as calcium carbonate, ferrous sulfate, zinc sulfate (ferric sulfate), and ferric sulfate (ferric sulfate) were added so that the mineral contents per ton of feed were 261.8 g of calcium, 183.3 g of Fe, 251.43 g of Zn, 16.18 g of Cu, , Copper sulfate, sodium selenite, manganese sulfate and calcium iodide) were further added to adjust the mineral composition to be the same.

상기 사료의 등록성분량은 조단백 17%, 조지방 2.5%, 칼슘 3.6%, 인 1.1%, 조섬유 5%, 조회분 15.5%, 메치오닌+시스틴+DL메치오닌수산화유도체 0.67%, 대사에너지 2,800kcal/kg이다. Metabolic energy of 2,800 kcal / kg was found in the diet of 17% crude protein, 2.5% crude fat, 1.1% phosphoric acid, 5% crude protein, 15.5% methionine + cystine + DL methionine hydroxide derivative and 0.67%

다. 실험군 배치All. Experiment group placement

각 실험군별 산란계의 배치수는 총 44수이다. 그 중 Fe-WPS, Fe-OCHT, Fe-GHAC, CaFe-GHAC, CaFe-IHAC, Ca-GHAC, 및 CaZn-GHAC 실험군 모두 산란계 5수이고, 살모넬라 비접종 대조군(정상군, CON1)은 3수, 실시예 1 내지 4 및 비교예 1 내지 3의 유기태화 미네랄 및 핵산 복합체가 첨가되지 않고 무기태 미네랄만으로 미네랄 조성을 맞춘 사료를 급이한 살모넬라 접종 대조군(CON2)은 6수로 설정하였다.The number of laying eggs per each experimental group is 44 in total. In the control group, the control group (normal group, CON1) was 3 times the number of the control group. In the control group, , The salmonella inoculation control group (CON2) fed with the minerals-adjusted feed without the organic minerals and nucleic acid complexes of Examples 1 to 4 and Comparative Examples 1 to 3 alone was set to six.

라. 1일 사료 섭이량la. Daily feed intake

1일 산란계 사료 섭이량은 산란계 1수당 120g공급하고, 하루 3회에 나누어 일정한 시간에 각 실험군에 맞는 식이를 제공하였다. 전체 시험기간 동안 항생제 무사용 조건이다.Daily diets were fed 120g per laying hens, divided into three times a day and fed to each experimental group at regular intervals. Antibiotic free conditions during the entire study period.

2) 실험일정2) Schedule of the experiment

실험군별로 각각 정해진 사료를 13일 동안 급이한 후 각 실험군별로 채혈을 하고, 생체중을 측정한 다음 Salmonella gallinarum 3001 균을 구강을 통해 공격접종하였다. 공격접종 후 닭의 외견상 감염증상이 나타나는지 매일 관찰하였다. 수양성설사가 심해지고 사료섭취율이 저하되며 조는 증상이 나타나는 공격접종 5일 후, 생체중을 측정하고 채혈을 한 후 모든 실험군을 부검하였다.Each feed group was fed for 13 days, and blood samples were collected for each group. Salmonella gallinarum 3001 was inoculated through the oral cavity. After the inoculation, the animals were observed daily for signs of apparent infection of the chickens. Five days after the attack, the live weight was measured and the blood was collected and all the experimental groups were autopsied.

3) 공격접종균주 배양 및 접종준비3) Cultivation of vaccination strain and preparation for inoculation

Salmonella gallinarum 3001(전염병실분양균주)을 1차로 brain hart infusion(BHI broth, BD BBL, USA) 10ml(x4개)에 12시간동안 진탕배양한 다음 OD(600nm)(1.2, 1.0, 0.8, 0.6)값을 측정하고 tryptic soy agar(TSA, BD Difco, USA)에 희석 도말하여 CFU/ml을 구했다(OD값 1.0일 때 균수가 0.6x109CFU/ml로 예상). 2차로 BHI broth 50ml(x4개)에 1:9 비율로 12시간동안 계대배양했다. 3차로 BHI broth 2L(50ml x 40개)에 1:9 비율로 12시간동안 계대배양했다. 배양된 균을 원심분리하여 균 pellet을 BHI broth 150ml에 현탁하여 수거했다. 산란계 1마리당 5ml(1x108CFU/마리/5ml)씩 존데를 이용하여 구강접종하였다. Salmonella gallinarum 3001 was first shake cultured in 10 ml (x4) of brain hart infusion (BHI broth, BD BBL, USA) for 12 hours and OD (600 nm) (1.2, 1.0, 0.8, 0.6) The value of CFU / ml was determined by diluting with tryptic soy agar (TSA, BD Difco, USA) (OD = 1.0, estimated to be 0.6x10 9 CFU / ml). Secondarily, 50 ml (x4 pieces) of BHI broth were subcultured in a 1: 9 ratio for 12 hours. And subcultured in a ratio of 1: 9 for 12 hours to 2 L of BHI broth (50 ml x 40). The cultured bacteria were centrifuged and the bacterial pellet was suspended in 150 ml of BHI broth. 5 ml (1x108 CFU / ml / 5 ml) per laying hens were orally inoculated using sonde.

4) 실험결과4) Experimental results

가. 폐사율end. Mortality rate

Salmonella gallinarum 공격접종 후부터 공격접종 5일 후 부검할 때까지 매일 실험군별 폐사한 닭의 마리수를 실험군별 마리수로 나누고 100을 곱하여 폐사율(%)을 구하여 표 2에 나타내었다. The number of dead birds per group was calculated by dividing the number of dead birds per experimental group by the number of birds per experimental group, and multiplied by 100, to calculate the mortality (%) from the day after the inoculation of Salmonella gallinarum until 5 days after the attack inoculation.

구분division 약어Abbreviation 4일차 폐사수4th day deadline 5일차 폐사수5th day 폐사율(%)Mortality (%) 비감염 대조군Uninfected control group CON1CON1 00 00 00 감염 대조군Infected Control CON2CON2 1One 00 2020 비교예1 Comparative Example 1 Fe-WPSFe-WPS 00 1One 2020 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 00 00 00 실시예1Example 1 Fe-GHACFe-GHAC 00 00 00 실시예2Example 2 CaFe-GHACCaFe-GHAC 00 00 00 실시예3Example 3 CaFe-IHACCaFe-IHAC 00 00 00 비교예3Comparative Example 3 Ca-GHACCa-GHAC 00 00 00 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 1One 00 2020

감염 대조군 및 비교예 4에서 공격접종 4일차에 닭 1마리 폐사하였고, 비교예 1에서는 5일차에 닭 1마리 폐사하여 폐사율이 20%이었고, 나머지 실험군에서는 공격접종 5일차까지는 폐사가 발생하지 않았다.In the infected control group and Comparative Example 4, one chicken died on the 4th day of the inoculation. In Comparative Example 1, the mortality rate was 20% and the mortality was not observed until the 5th day of the inoculation in the remaining experimental group.

상기 5일차까지 살아있는 닭만을 대상으로 이후 실험을 실시하였다.Experiments were conducted on only live chicken until the 5th day.

나. 사료섭취량I. Feed intake

사료섭취량은 살모넬라 접종 전후의 총기간, 살모넬라 접종 전 및 살모넬라 접종 후로 구분하여, 각 실험군별로 전날 급이한 사료 무게에서 다음날 오전 8시에 남은 사료의 무게를 뺀 실제 급이량의 무게를 측정하고 각 실험군 마리수로 나누어 사료섭취량(g/day chicken)을 계산하여 표 3에 나타내었다.Feed intake was determined by dividing the total period before and after inoculation of salmonella, before salmonella inoculation, and after inoculation with salmonella, and weighed the actual feed weight of each experimental group minus the weight of the remaining feed at 8 o'clock the following day The feed intake (g / day chicken) was divided by the number of mariners in each experimental group and is shown in Table 3.

구분division 약어Abbreviation 총기간Total duration 접종 전Before vaccination 접종 후After inoculation 접종전후 섭취량차이Difference of intake before and after inoculation 비감염 대조군Uninfected control group CON1CON1 95.4±11.895.4 ± 11.8 93.9±12.993.9 ± 12.9 99.5±8.199.5 ± 8.1 5.65.6 감염 대조군Infected Control CON2CON2 105.5±16.8105.5 ± 16.8 110.7±4.5110.7 ± 4.5 91.4±29.991.4 ± 29.9 -19.3-19.3 비교예1 Comparative Example 1 Fe-WPSFe-WPS 99.7±24.999.7 ± 24.9 104.9±7.7104.9 ± 7.7 85.3±48.285.3 ± 48.2 -19.6-19.6 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 95.5±22.795.5 ± 22.7 105.1±9.2105.1 ± 9.2 68.9±28.868.9 ± 28.8 -36.2-36.2 실시예1Example 1 Fe-GHACFe-GHAC 103.9±7.6103.9 ± 7.6 104.3±6.5104.3 ± 6.5 103.4±8.8103.4 ± 8.8 -0.9-0.9 실시예2Example 2 CaFe-GHACCaFe-GHAC 101.1±7.4101.1 + - 7.4 100.8±7.0100.8 ± 7.0 102.1±9.5102.1 ± 9.5 1.31.3 실시예3Example 3 CaFe-IHACCaFe-IHAC 102.7±8.8102.7 ± 8.8 102.5±8.2102.5 ± 8.2 102.9±9.4102.9 ± 9.4 0.40.4 비교예3Comparative Example 3 Ca-GHACCa-GHAC 97.7±13.497.7 ± 13.4 100.1±7.2100.1 ± 7.2 91.0±24.291.0 + - 24.2 -9.1-9.1 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 97.5±20.097.5 ± 20.0 104.5±8.0104.5 ± 8.0 87.4±31.387.4 ± 31.3 -17.1-17.1

비감염 대조군과 실시예 1 내지 4에서는 접종 전과 접종 후의 섭취량에 차이가 거의 없거나 접종 후의 섭취량이 오히려 증가하였다. 그러나 감염 대조군에서는 사료 섭취량이 19.6 g 감소하였고, 비교예 1 및 비교예 4에서는 이와 비슷한 수준의 섭취량 감소를 나타내었고, 비교예 2에서는 감염 대조군의 2배에 달하는 섭취량 감소를 나타내었다.In the uninfected control group and in Examples 1 to 4, there was almost no difference in the intake before and after the inoculation, or the intake after the inoculation was rather increased. However, in the infected control group, the feed intake was decreased by 19.6 g, and the intake level of Comparative Example 1 and Comparative Example 4 was reduced to a similar level. In Comparative Example 2, the intake level was decreased to twice that of the infected control group.

다. 산란율 및 파란율All. Egg Production Rate and Blue Rate

매일 낮 12시에 산란한 계란을 수거하여 각 실험군별 계란수와 파란수를 파악하고 군별 마리수로 나눈 다음 100을 곱하여 일일 산란율(%) 및 파란율(%)을 구하여 각각 표 4 및 표 5에 나타내었다.The egg laying at 12 o'clock every day was collected, and the number of eggs and the number of blue eggs were determined for each experimental group, divided by the number of eggs per group, and then multiplied by 100 to obtain a daily egg production rate (%) and a blue egg rate (% Respectively.

구분division 약어Abbreviation 총기간Total duration 접종 전Before vaccination 접종 후After inoculation 비감염 대조군Uninfected control group CON1CON1 80.480.4 77.877.8 86.786.7 감염 대조군Infected Control CON2CON2 85.385.3 87.587.5 80.080.0 비교예1 Comparative Example 1 Fe-WPSFe-WPS 67.167.1 63.363.3 76.076.0 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 83.583.5 91.791.7 64.064.0 실시예1Example 1 Fe-GHACFe-GHAC 8.358.35 84.384.3 81.881.8 실시예2Example 2 CaFe-GHACCaFe-GHAC 83.583.5 83.383.3 84.084.0 실시예3Example 3 CaFe-IHACCaFe-IHAC 83.083.0 82.482.4 84.284.2 비교예3Comparative Example 3 Ca-GHACCa-GHAC 75.375.3 75.075.0 76.076.0 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 78.878.8 80.080.0 76.076.0

구분division 약어Abbreviation 총기간Total duration 접종 전Before vaccination 접종 후After inoculation 비감염 대조군Uninfected control group CON1CON1 0.00.0 0.00.0 0.00.0 감염 대조군Infected Control CON2CON2 1.01.0 0.00.0 3.33.3 비교예1 Comparative Example 1 Fe-WPSFe-WPS 1.21.2 0.00.0 4.04.0 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 2.42.4 0.00.0 8.08.0 실시예1Example 1 Fe-GHACFe-GHAC 0.40.4 0.00.0 1.21.2 실시예2Example 2 CaFe-GHACCaFe-GHAC 0.00.0 0.00.0 0.00.0 실시예3Example 3 CaFe-IHACCaFe-IHAC 0.30.3 0.00.0 0.80.8 비교예3Comparative Example 3 Ca-GHACCa-GHAC 7.07.0 6.76.7 8.08.0 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 3.53.5 3.33.3 4.04.0

비감염 대조군은 산란율이 접종 후 오히려 증가하고, 파란율에 변화가 없었으나, 감염 대조군은 접종 후 산란율이 7.5% 감소하고, 파란율은 3.3% 증가하였다.In the uninfected control group, the egg production rate was increased after inoculation and the blue ratio was not changed. In the infected control group, the egg production rate was decreased by 7.5% and the blue ratio was increased by 3.3%.

비교예 1에서는 접종 후 산란율은 다소 증가하였으나, 접종 전 산란율이 특이하게 낮은 경우였으며, 접종 후 파란율은 4% 증가하였다. 비교예 2에서는 접종 후 산란율이 27.7% 현저히 감소하고, 파란율도 8%나 증가하였다.In Comparative Example 1, the egg production rate after inoculation was slightly increased, but the pre-inoculation rate was unusually low, and the after-inoculation rate increased by 4%. In Comparative Example 2, the egg production rate after inoculation was remarkably decreased by 27.7% and the blue ratio was increased by 8%.

실시예 1 내지 3에서는 접종 후 산란율이 0.7 내지 1.8% 증가하거나(실시예 2 및 3) 감소하더라도 그 폭이 매우 미미하였고, 파란율도 0 내지 1.2% 범위 이내였다.In Examples 1 to 3, even when the egg production rate after the inoculation was increased by 0.7 to 1.8% (Examples 2 and 3), the width was very small and the blue ratio was also within the range of 0 to 1.2%.

비교예 3에서는 접종 후 산란율이 1% 증가하고, 파란율은 1.3%감소하였고, 비교예 4에서는 접종 후 산란율이 4% 감소하고, 파란율은 0.7% 감소하였다.In Comparative Example 3, the egg production rate was increased by 1% and the blue ratio was decreased by 1.3%. In Comparative Example 4, the egg production rate was decreased by 4% and the blue ratio was decreased by 0.7%.

라. 공격접종 후 체중 변화la. Weight change after inoculation

공격접종 전과 접종 5일 후 생체중(g)을 측정하여, 공격접종 전과 비교했을 때의 접종 5일차 체중 증감율(%)을 표 6에 나타내었다.The body weight (g) was measured before the attack and 5 days after the inoculation, and the weight increase rate (%) of the inoculation on the 5th day after inoculation was shown in Table 6.

구분division 약어Abbreviation 공격접종 전 체중(g)Weight before attack (g) 접종 5일차 체중(g)Inoculation Day 5 Body weight (g) 체중증감율(%)Weight increase rate (%) 비감염 대조군Uninfected control group CON1CON1 1522.0±167.11522.0 占 167.1 1544.7±174.891544.7 ± 174.89 101.5101.5 감염 대조군Infected Control CON2CON2 1905.7±155.31905.7 ± 155.3 1741.0±125.01741.0 + - 125.0 91.491.4 비교예1 Comparative Example 1 Fe-WPSFe-WPS 1843.2±241.91843.2 + 241.9 1636.4±213.61636.4 ± 213.6 88.888.8 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 1761.2±213.31761.2 ± 213.3 1580.8±185.61580.8 ± 185.6 89.889.8 실시예1Example 1 Fe-GHACFe-GHAC 1685.4±168.81685.4 ± 168.8 1582.9±145.61582.9 ± 145.6 93.993.9 실시예2Example 2 CaFe-GHACCaFe-GHAC 1598.0±34.31598.0 + - 34.3 1493.6±59.41493.6 ± 59.4 93.593.5 실시예3Example 3 CaFe-IHACCaFe-IHAC 1724.5±63.21724.5 ± 63.2 1623.4±207.81623.4 ± 207.8 94.194.1 비교예3Comparative Example 3 Ca-GHACCa-GHAC 1730.0±220.51730.0 + - 220.5 1596.8±227.51596.8 ± 227.5 92.392.3 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 1767.2±152.71767.2 ± 152.7 1602.8±86.61602.8 + - 86.6 90.790.7

비감염 대조군에서만 체중이 증가하였고, 감염 대조군에서는 체중이 8.6% 감소하였다. 비교예 1, 비교예 2 및 비교예 4에서는 상기 감염 대조군에 비해서 체중이 더 큰 폭으로 감소하였고, 실시예 1 내지 3에서는 감염 대조군에 비해서 체중 감소의 폭이 더 작았다.Body weight was increased only in the non-infected control group and 8.6% in the infected control group. In Comparative Example 1, Comparative Example 2 and Comparative Example 4, the body weight decreased more than the infected control group, and in Examples 1 to 3, the weight loss was smaller in comparison with the infected control group.

마. 비장종대hemp. Spleen enlargement

공격접종 5일 후 부검하여 비장의 무게, 높이/너비를 구하여 비장종대의 정도를 실험군별로 비교하여 표 7에 나타내었다. Five days after the attack, the spleen's weight and height / width were determined and the degree of splenic enlargement was compared according to the experimental groups.

구분division 약어Abbreviation 무게(g)Weight (g) 높이(mm)Height (mm) 너비(mm)Width (mm) 비감염 대조군Uninfected control group CON1CON1 1.6±0.31.6 ± 0.3 19.4±0.719.4 ± 0.7 12.2±1.112.2 ± 1.1 감염 대조군Infected Control CON2CON2 4.7±1.54.7 ± 1.5 26.3±3.026.3 ± 3.0 17.9±2.617.9 ± 2.6 비교예1 Comparative Example 1 Fe-WPSFe-WPS 5.1±1.65.1 ± 1.6 26.6±2.726.6 ± 2.7 19.7±2.519.7 ± 2.5 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 3.7±1.53.7 ± 1.5 23.9±3.423.9 ± 3.4 17.6±3.117.6 ± 3.1 실시예1Example 1 Fe-GHACFe-GHAC 2.8±1.62.8 ± 1.6 22.9±3.822.9 ± 3.8 15.7±3.215.7 ± 3.2 실시예2Example 2 CaFe-GHACCaFe-GHAC 2.6±1.62.6 ± 1.6 22.9±4.422.9 ± 4.4 15.3±3.815.3 ± 3.8 실시예3Example 3 CaFe-IHACCaFe-IHAC 2.7±1.82.7 ± 1.8 23.1±3.523.1 ± 3.5 16.1±2.716.1 ± 2.7 비교예3Comparative Example 3 Ca-GHACCa-GHAC 4.0±3.04.0 ± 3.0 24.8±3.224.8 ± 3.2 17.9±5.117.9 ± 5.1 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 4.2±1.94.2 ± 1.9 25.0±5.525.0 ± 5.5 17.9±2.717.9 ± 2.7

살모넬라 감염으로 인하여 비감염 대조군에 비하여 감염 대조군에서 비장 종대가 발생하였으나, 실시예 1 내지 3에서 비장의 무게와 크기가 감염 대조군에 비해 감소하였고, 비교예 중에서는 비교예 2에서 상대적으로 무게와 크기가 감염 대조군에 비해 감소되었다.The spleen weight and size of the spleen in Examples 1 to 3 were reduced compared to the infected control group, although spleen enlargement occurred in the infected control group compared to the non-infected control group due to Salmonella infection. In Comparative Example 2, Compared with the control group.

바. 바닥 분변 내 살모넬라 균수bar. Salmonella bacteria in the bottom feces

공격접종 4일차에 계사바닥에 톱밥을 깔고 그 위에 멸균 paper towel을 깔았다. 3~4시간 뒤에 한 실험군에 3군데에서 각각 멸균 스푼으로 바닥분변을 채취하고, 즉시 0.1g을 무게를 재고 나머지는 바로 -80℃ deep freezer에 보관한 후, 살모넬라균수를 측정하여 표 8에 나타내었다. 감염 대조군(CON2)를 기준으로 상대적인 균수를 백분율로 나타내어 증감율(%)을 기재하였다.On the 4th day of attack, sawdust was laid on the floor of the house and a sterilized paper towel was laid on it. Three to four hours later, the bottom feces were collected from each of the three experimental sites by using a sterilized spoon. Immediately, 0.1 g was weighed and the rest was stored in a deep-freezer at -80 ° C. and the number of Salmonella bacteria was measured. . The percent increase (%) was expressed as percentage relative to the number of infectious control (CON2).

구분division 약어Abbreviation 균수(CFU/ml)Number of bacteria (CFU / ml) 증감율(%)Rate of increase / decrease (%) 비감염 대조군Uninfected control group CON1CON1 00 -- 감염 대조군Infected Control CON2CON2 1.57x109 1.57x10 9 100.0100.0 비교예1 Comparative Example 1 Fe-WPSFe-WPS 1.85x109 1.85x10 9 118.0118.0 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 4.46x108 4.46x10 8 28.428.4 실시예1Example 1 Fe-GHACFe-GHAC 5.56x105 5.56x10 5 0.00.0 실시예2Example 2 CaFe-GHACCaFe-GHAC 1.50x106 1.50x10 6 0.10.1 실시예3Example 3 CaFe-IHACCaFe-IHAC 1.12x106 1.12x10 6 0.10.1 비교예3Comparative Example 3 Ca-GHACCa-GHAC 2.04x109 2.04x10 9 129.7129.7 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 7.06x109 7.06x10 9 449.3449.3

살모넬라를 공격 접종한 감염 대조군에서는 살모넬라 균수가 109 수준으로 검출되었고, 비교예 1, 비교예 3 및 비교예 4에서는 살모넬라 균수가 감소하였고, 비교예 2 및 실시예 1 내지 3에서는 살모넬라 균수가 감소하였다. 특히 실시예 1 내지 3에서는 3 로그 사이클 이상으로 현저히 감소시켰다. Salmonella bacteria counts were detected at 10 9 levels in the infected control group attacked with Salmonella. In Comparative Example 1, Comparative Example 3 and Comparative Example 4, the number of Salmonella bacteria was decreased. In Comparative Example 2 and Examples 1 and 3, Respectively. Especially in Examples 1 to 3, it was significantly reduced to 3 log cycles or more.

사. T세포 분포 및 사이토카인 함량four. T cell distribution and cytokine content

공격접종 전과 접종 5일 후 2번 채혈을 하여 전혈과 혈청을 분리하였다. 전혈에서 T 세포 분포를 확인하여 표 9에 나타내었다. Blood was collected twice before and 5 days after inoculation and whole blood and serum were separated. The distribution of T cells in whole blood was confirmed and shown in Table 9.

상기 전혈은 3ml을 같은 부피의 PBS로 희석하였다. 희석된 혈액시료는 3ml의 LymphoprepTM(Axis-Shield, Oslo, Norway)에 적용시키고 상온 1,300rpm에서 30분간 원심분리하여 PBMC간의 결합을 분리하였다. PBMC는 PBS로 1회 washing한 뒤, 상층액을 제거하고 RBC lysis buffer 1ml로 부유시켜서 상온에서 10분간 RBC를 분해하였다. 2,000rpm에서 10분간 원심분리하여 상층액을 제거하고 PBS 10ml로 부유시켜 washing하였다. 1ml PBS/1% BSA로 cell을 부유시켜 다시 한번 washing하였다. cell을 counting하여 1x108cell/ml로 맞췄다. 그리고 분리된 세포의 T helper cell(CD3+ CD4+)과 T cytotoxic cell(CD3+ CD8+)의 비율을 분석하였다. T helper cell은 CD3와 FITC-conjugated mouse anti chicken CD4로 염색하여 구분하였다. T cytotoxic cell은 CD3와 PRE-conjugated mouse anti chicken CD8으로 염색하여 구분하였다.The whole blood was diluted with 3 ml of the same volume of PBS. The diluted blood samples were applied to 3 ml of Lymphoprep (Axis-Shield, Oslo, Norway) and centrifuged at 1,300 rpm for 30 minutes at room temperature to separate the binding between the PBMCs. PBMCs were washed once with PBS, and then the supernatant was removed and suspended in 1 ml of RBC lysis buffer, and the RBC was digested at room temperature for 10 minutes. After centrifugation at 2,000 rpm for 10 minutes, the supernatant was removed and suspended in 10 ml of PBS. Cells were suspended in 1 ml PBS / 1% BSA and washed again. Cells were counted and adjusted to 1x108 cells / ml. The ratio of T helper cell (CD3 + CD4 +) and T cytotoxic cell (CD3 + CD8 +) in isolated cells was analyzed. T helper cells were stained with CD3 and FITC-conjugated mouse anti-CD4. T cytotoxic cells were stained with CD3 and PRE-conjugated mouse anti-CD8.

96well plate well안에 cell 100㎕와 antibody CD3-CD4-FITC, CD3-CD8-RPE, CD3-CD4-FITC-CD8-PRE 1㎕로 염색하였다. 알루미늄 호일로 plate를 둘러싸서 어두운 곳에서 실온으로 30분간 배양하였다. 원심분리하여 상층액을 제거하고 PBS로 washing하였다. 그리고 fixing buffer로 cell을 고정한 다음 유세포분석기를 이용하여 분석하였으며, Lymphocyte는 FSC/SSC를 이용하여 FITC와 PE 모두 염색되었는지 여부를 10,000건 분석하였다. 각각의 림프구에 대한 결과는 FSC/SSC Lymphocyte gate에 백분율로 표시하였다.The cells were stained with 100 μl of the cells in 96-well plate wells and 1 μl of antibody CD3-CD4-FITC, CD3-CD8-RPE and CD3-CD4-FITC-CD8-PRE. The plate was wrapped with aluminum foil and incubated in the dark at room temperature for 30 minutes. The supernatant was removed by centrifugation and washed with PBS. The cells were fixed with a fixing buffer and analyzed by flow cytometry. Lymphocytes were analyzed by FSC / SSC for 10,000 staining of FITC and PE. The results for each lymphocyte were expressed as a percentage in the FSC / SSC lymphocyte gate.

구분division 약어Abbreviation 살모넬라Salmonella 접종 전Before vaccination 살모넬라Salmonella 접종 후After inoculation CD3+/CD4+CD3 + / CD4 + CD3+/CD8+CD3 + / CD8 + CD3+/CD4+CD3 + / CD4 + CD3+/CD8+CD3 + / CD8 + 비감염 대조군Uninfected control group CON1CON1 3.62±0.943.62 + - 0.94 2.89±1.272.89 ± 1.27 2.91±0.9692.91 0.969 1.00±0.711.00 0.71 감염 대조군Infected Control CON2CON2 3.62±0.943.62 + - 0.94 2.89±1.272.89 ± 1.27 1.19±0.191.19 ± 0.19 0.44±0.570.44 ± 0.57 비교예1 Comparative Example 1 Fe-WPSFe-WPS 5.10±0.225.10 ± 0.22 3.65±0.733.65 ± 0.73 1.44±0.131.44 ± 0.13 0.45±0.470.45 + 0.47 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 3.72±1.333.72 ± 1.33 2.84±1.632.84 ± 1.63 1.48±0.731.48 ± 0.73 0.26±0.210.26 ± 0.21 실시예1Example 1 Fe-GHACFe-GHAC 3.15±0.083.15 ± 0.08 2.32±1.252.32 ± 1.25 3.21±1.223.21 ± 1.22 0.68±0.210.68 ± 0.21 실시예2Example 2 CaFe-GHACCaFe-GHAC 2.81±0.042.81 + 0.04 2.05±0.872.05 ± 0.87 3.11±1.573.11 ± 1.57 0.74±0.240.74 + 0.24 실시예3Example 3 CaFe-IHACCaFe-IHAC 3.25±0.523.25 ± 0.52 2.15±1.562.15 ± 1.56 3.33±1.123.33 ± 1.12 0.83±0.550.83 + - 0.55 비교예3Comparative Example 3 Ca-GHACCa-GHAC 3.61±2.363.61 ± 2.36 2.28±2.842.28 ± 2.84 1.67±0.461.67 + - 0.46 0.97±0.560.97 + - 0.56 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 5.56±3.625.56 ± 3.62 4.19±3.334.19 ± 3.33 1.25±0.901.25 0.90 0.22±0.100.22 0.10

살모넬라 감염에 의해 CD3+/CD4+ T세포와 CD3+/CD8+ T세포의 분포가 감염 대조군에서 현저히 감소하였다. 비교예 1 내지 4에서는 CD3+/CD4+ T세포가 분포가 감염 대조군과 큰 차이가 없었으나 실시예 1 내지 3에서는 CD3+/CD4+ T세포를 현저히 활성화시켰다. 또한 비교예 1, 2 및 4에서는 CD3+/CD4+ T세포가 분포가 감염 대조군과 큰 차이가 없거나 오히려 감소시켰으나, 실시예 1 내지 3 및 비교예 3에서는 CD3+/CD4+ T세포를 현저히 활성화시켰다.Distribution of CD3 + / CD4 + T cells and CD3 + / CD8 + T cells by Salmonella infection was significantly reduced in the infected control group. In Comparative Examples 1 to 4, distribution of CD3 + / CD4 + T cells was not significantly different from that in the infected control, but in Examples 1 to 3, CD3 + / CD4 + T cells were significantly activated. In addition, in Comparative Examples 1, 2 and 4, the distribution of CD3 + / CD4 + T cells was not significantly different from that of the infected control group, but rather, the CD3 + / CD4 + T cells were significantly activated in Examples 1 to 3 and Comparative Example 3.

상기 혈청에서 IL-2 및 IFN-γ 농도(ng/ml)를 ELISA방법으로 측정하여 표 10에 나타내었다.The concentrations of IL-2 and IFN-y (ng / ml) in the serum were measured by ELISA and are shown in Table 10.

구분division 약어Abbreviation 살모넬라Salmonella 접종 전Before vaccination 살모넬라Salmonella 접종 후After inoculation IL-2IL-2 IFN-γIFN-y IL-2IL-2 IFN-γIFN-y 비감염 대조군Uninfected control group CON1CON1 51.95±18.5951.95 ± 18.59 35.87±4.2635.87 + - 4.26 52.29±13.3752.29 + 13.37 33.21±6.4133.21 + - 6.41 감염 대조군Infected Control CON2CON2 33.38±7.8933.38 + - 7.89 29.46±4.6129.46 + - 4.61 38.95±7.6938.95 + - 7.69 31.50±3.2431.50 ± 3.24 비교예1 Comparative Example 1 Fe-WPSFe-WPS 41.07±18.4341.07 ± 18.43 29.96±3.3629.96 ± 3.36 54.89±7.9554.89 ± 7.95 36.34±5.2536.34 ± 5.25 비교예2 Comparative Example 2 Fe-OCHTFe-OCHT 33.17±13.9533.17 + - 13.95 29.86±3.7929.86 ± 3.79 34.66±4.6034.66 + - 4.60 29.82±3.4929.82 ± 3.49 실시예1Example 1 Fe-GHACFe-GHAC 38.24±12.2438.24 + - 12.24 30.66±2.8730.66 + 2.87 58.42±4.3058.42 + - 4.30 35.51±3.4735.51 + - 3.47 실시예2Example 2 CaFe-GHACCaFe-GHAC 40.47±1.4640.47 + 1.46 32.54±3.9632.54 ± 3.96 61.25±35.7161.25 + - 35.71 36.88±7.3736.88 + - 7.37 실시예3Example 3 CaFe-IHACCaFe-IHAC 39.85±11.2539.85 + - 11.25 32.12±3.3632.12 ± 3.36 55.68±23.4555.68 ± 23.45 35.68±6.5535.68 + - 6.55 비교예3Comparative Example 3 Ca-GHACCa-GHAC 43.25±11.4943.25 + - 11.49 31.79±7.6531.79 ± 7.65 43.96±5.5443.96 + - 5.54 38.52±6.7538.52 + - 6.75 비교예4Comparative Example 4 CaZn-GHACCaZn-GHAC 37.99±9.7937.99 ± 9.79 30.28±4.6430.28 + - 4.64 43.04±10.7643.04 + - 10.76 35.76±5.2135.76 ± 5.21

살모넬라 감염에 의해 감염 대조군에서 IL-2 및 인터페론-γ의 분비가 다소 증가하는 경향을 나타내었다. 그러나 비교예 2에서는 접종 전후에 IL-2 및 인터페론-γ의 분비량에 차이를 나타내지 않았다. 비교예 3에서는 접종 전후에 IL-2의 분비량에는 차이가 없었으나, 인터페론-γ의 분비는 증가시켰다. 비교예 1, 비교예 4 및 실시예 1 내지 3에서는 감염 후 IL-2 및 인터페론-γ의 분비를 현저히 증가시켰다.The secretion of IL-2 and interferon-y in the infected control group was slightly increased by Salmonella infection. However, in Comparative Example 2, there was no difference in the amount of IL-2 and interferon-y secretion before and after inoculation. In Comparative Example 3, there was no difference in the secretion amount of IL-2 before and after inoculation, but the secretion of interferon-γ was increased. In Comparative Example 1, Comparative Example 4 and Examples 1 to 3, secretion of IL-2 and interferon-y after infection was significantly increased.

5) 통계처리5) Statistical processing

실험 결과 데이터는 평균±표준편차(SD)로 표시하였다. 수치 분석은 Student t-test를 통해 수행하였다. 모든 데이터 분석은 SPSS를 이용하였다. P<0.05를 유의성 있는 수준으로 평가하였다.The experimental data were expressed as mean ± standard deviation (SD). Numerical analysis was performed by Student t-test. All data were analyzed using SPSS. P <0.05 was evaluated as significant.

Claims (4)

유기태화 철 및 퓨린염기 함유 핵산 단량체의 복합체를 유효성분으로 하는 살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물에 있어서,
상기 퓨린염기 함유 핵산 단량체는 이노신산 또는 구아닐산인 것을 특징으로 하는 살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물.
A feed additive composition containing an organic minerals complex for preventing or treating salmonella diarrhea comprising a complex of an organotissive iron and a purine base-containing nucleic acid monomer as an active ingredient,
Wherein the purine base-containing nucleic acid monomer is inosine or guanylic acid.
삭제delete 제1항에 있어서, 상기 유기태화 철 및 퓨린염기 함유 핵산 단량체의 복합체는 칼슘, 아연, 마그네슘, 칼륨, 구리, 셀렌, 크롬, 몰리브덴 및 요오드 중에서 선택되는 어느 하나 또는 둘 이상의 미네랄이 추가로 킬레이트된 유기태화 미네랄 및 퓨린염기 함유 핵산 단량체의 복합체인 것을 특징으로 하는 살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물.
The complex of the organotin ferrous and purine base-containing nucleic acid monomers according to claim 1, wherein one or more of the minerals selected from calcium, zinc, magnesium, potassium, copper, selenium, chromium, molybdenum and iodine is further chelated Wherein the feed additive composition is a complex of an organic minerals and a nucleic acid monomer containing a purine base.
제3항에 있어서, 상기 유기태화 미네랄 및 퓨린염기 함유 핵산 단량체의 복합체는 유기태화 칼슘과 철 및 퓨린염기 함유 핵산 단량체의 복합체이고, 상기 칼슘과 철분의 중량비는 5:1 내지 1:5인 것을 특징으로 하는 살모넬라성 설사 예방 또는 치료용 유기태화 미네랄 복합체 함유 사료 첨가제 조성물.4. The method according to claim 3, wherein the complex of the organotinized mineral and the purine base-containing nucleic acid monomer is a complex of calcium oxalate and a nucleic acid monomer containing iron and purine base, and the weight ratio of calcium to iron is from 5: 1 to 1: 5 Wherein the feed additive composition comprises an organotin mineral complex for preventing or treating Salmonella diarrhea.
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JP2003088333A (en) 2001-09-14 2003-03-25 Bizen Chemical Co Ltd Method for producing essence containing organic mineral in high concentration
CN105831449A (en) 2016-04-12 2016-08-10 南京可莱威饲料有限公司 Concentrate feed containing chitosan oligosaccharide for weaning pig and preparation method thereof

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JP2003088333A (en) 2001-09-14 2003-03-25 Bizen Chemical Co Ltd Method for producing essence containing organic mineral in high concentration
CN105831449A (en) 2016-04-12 2016-08-10 南京可莱威饲料有限公司 Concentrate feed containing chitosan oligosaccharide for weaning pig and preparation method thereof

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210066394A (en) 2019-11-28 2021-06-07 건국대학교 산학협력단 Feed additive composition containing lactobacillus kefiranofaciens dn1

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