KR20200001408A - Composition of feed stuff for sea cucumber containing asthma improving substance and manufacturing method thereof - Google Patents

Composition of feed stuff for sea cucumber containing asthma improving substance and manufacturing method thereof Download PDF

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KR20200001408A
KR20200001408A KR1020180074340A KR20180074340A KR20200001408A KR 20200001408 A KR20200001408 A KR 20200001408A KR 1020180074340 A KR1020180074340 A KR 1020180074340A KR 20180074340 A KR20180074340 A KR 20180074340A KR 20200001408 A KR20200001408 A KR 20200001408A
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sea cucumber
feed
sea
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KR102276217B1 (en
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강석중
정우철
엠디 아니수자만
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경상대학교산학협력단
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/20—Animal feeding-stuffs from material of animal origin
    • A23K10/26—Animal feeding-stuffs from material of animal origin from waste material, e.g. feathers, bones or skin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
    • 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
    • A23K20/142—Amino acids; Derivatives thereof
    • A23K20/147—Polymeric derivatives, e.g. peptides or proteins
    • 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
    • A23K20/174—Vitamins
    • 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
    • A23K20/26—Compounds containing phosphorus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
    • A23K30/20—Dehydration
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish
    • Y02A40/818—Alternative feeds for fish, e.g. in aquacultures

Abstract

The present invention relates to a composition for Apostichopus japonicus culture feed, and more particularly provides a composition for Apostichopus japonicus culture feed containing a marine microalgae powder as an active component which has an enhanced asthma alleviation effect, and a manufacturing method thereof.

Description

천식 개선물질을 포함하는 해삼사료 조성물 및 그 제조방법{Composition of feed stuff for sea cucumber containing asthma improving substance and manufacturing method thereof}Composition of feed stuff for sea cucumber containing asthma improving substance and manufacturing method

본 발명은 천식 개선물질을 포함하는 해삼사료 조성물에 관한 것으로서, 더 상세하게는 천식 개선물질을 포함하는 해삼사료 조성물 및 그 제조방법에 관한 것이다.The present invention relates to a sea cucumber feed composition comprising an asthma improving material, and more particularly to a sea cucumber feed composition comprising an asthma improving material and a method of manufacturing the same.

해삼(Apostichopus japonicus)은 중국에서 고가의 식품이지만, 최근 해양오염과 연안매립으로 인한 서식지 파괴 때문에 중국내 생산량이 부족하여 수요를 충족시키기 위하여 한국, 일본 및 동남아 지역에서 대량수입에 의존하고 있다. 그래서 우리나라에서도 수산물 수출 10대 전략품종의 하나로 지정하여 해삼종자생산기슬개발, 가공기술개발과 함께 해삼클러스터조성사업 등 해삼양식에 박차를 가하고 있다. 특히 해삼은 중국에서 예로부터 관절염, 고혈압, 상처치유 및 외과수술 후 건강회복에 사용해 왔으며, 천식질환 개선에도 효과가 있는 것으로 알려져 있다. 따라서 최근 해삼의 부가가치를 높이기 위하여 양식해삼을 단순 가공하여 수출하는 형태에서 벗어나 기능성을 갖는 고부가의 해삼생산이 요구되고 있다.Sea cucumber ( Apostichopus japonicus ) is an expensive food in China, but due to marine pollution and habitat destruction due to coastal reclamation, the production of China has been insufficient, so it relies on mass imports from Korea, Japan and Southeast Asia to meet the demand. Therefore, in Korea, it is designated as one of the top 10 strategic varieties of seafood exports, and is accelerating sea cucumber culture such as sea cucumber seed production chain development, processing technology development, and sea cucumber cluster composition business. In particular, sea cucumber has been used in China to restore health after arthritis, hypertension, wound healing, and surgery, and is known to be effective in improving asthma disease. Therefore, in order to increase the added value of sea cucumber recently, the production of high value sea cucumber with the functionality beyond the form of simple processing and exporting sea cucumber is required.

한편 천식(asthma)은 알레르기성 호흡기 질환의 일종으로 세계적으로 약 3억 명 인구가 고통을 겪고 있으며, 이로 인한 사망자는 연간 약 25만 명으로 그 숫자는 계속 증가하고 있다. 또한 천식은 간접흡연, 미세먼지, 황사, 식습관 및 어린이 면역체계 약화 등 요인으로 발병연령이 점차 낮아지는 양상을 나타내어 세계보건기구(WHO)의 관심이 증대되고 있는 질환중의 하나이다. 이와 관련하여 대한민국 공개특허 제2013-0009425호는 친환경 소재를 이용한 해삼 사료용 조성물 및 이의 제조방법에 대해 개시하고 있다. Asthma is a type of allergic respiratory disease that affects about 300 million people worldwide, resulting in an estimated 250,000 deaths per year. In addition, asthma is one of the diseases in which the World Health Organization (WHO) is increasing interest due to the gradually decreasing age due to factors such as indirect smoking, fine dust, yellow dust, eating habits, and weakening the immune system of children. In this regard, the Republic of Korea Patent Publication No. 2013-0009425 discloses a composition for sea cucumber feed using an environmentally friendly material and a manufacturing method thereof.

그러나 상기 선행기술은 해조류를 이용하여 고형화 형태의 해삼 사료를 제조하는 것으로 단순히 급여에 의한 양적증가만을 고려할 뿐 건강식품으로서 기능성을 나타내는 양식 해삼 생산을 위한 사료용 조성물이 아니다. However, the prior art is to prepare a sea cucumber feed of the solidified form using seaweed, it is not merely a feed composition for production of cultured sea cucumber, which merely considers the quantitative increase by salary, and shows functionality as a health food.

본 발명은 상기와 같은 문제점을 해결하기 위한 것으로서, 종래 해삼 양식방법과 비교하여 성장이 우수하고 천식 개선물질을 다량 포함하는 해삼을 생산하기 위한 사료 조성물 및 그 제조방법을 제공하는 것을 목적으로 한다. 그러나 이러한 과제는 예시적인 것으로, 이에 의해 본 발명의 범위가 한정되는 것은 아니다.The present invention is to solve the above problems, and to provide a feed composition and a method of producing the same for producing a sea cucumber excellent growth compared to conventional sea cucumber culture method and containing a large amount of asthma improving material. However, these problems are exemplary, and the scope of the present invention is not limited thereby.

본 발명의 일 관점에 따르면, 갈파래, 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상 조류의 분말 10 내지 20 중량부, 지충이(Sargassum thunbergii) 10 내지 20 중량부, 대두박(soybean meal) 5 내지 10 중량부, 조개분말(shell fish powder) 5 내지 10 중량부, 패각분말(Shell powder) 1 내지 3 중량부, 인산칼슘(calcium phosphate) 1 내지 3 중량부, 효모 추출물(yeast extract) 3 내지 8 중량부, 콩 레시틴(soyabean lecithin) 2 내지 6 중량부, 미네랄(mineral) 0.2 내지 0.7 중량부, 비타민(vitamin) 0.2 내지 0.7 중량부, 및 바다 진흙(sea mud) 20 내지 60 중량부를 포함하는, 천식 개선 효과가 강화된 해삼의 생산을 위한 해삼 양식용 사료조성물이 제공된다. According to an aspect of the present invention, 10 to 20 parts by weight of one or two or more algae selected from the group consisting of brown seaweed, kelp and Nanclochloros s sp., 10 to 20 parts by weight, Sargassum thunbergii 20 parts by weight, 5-10 parts by weight of soybean meal, 5-10 parts by weight of shell fish powder, 1-3 parts by weight of shell powder, 1-3 parts by weight of calcium phosphate Parts, 3 to 8 parts by weight of yeast extract, 2 to 6 parts by weight of soyabean lecithin, 0.2 to 0.7 parts by weight mineral, 0.2 to 0.7 parts by weight vitamin, and sea mud ( Sea mud) Provided with a sea cucumber culture feed composition for the production of sea cucumber enhanced asthma-improving effect, including 20 to 60 parts by weight.

본 발명의 다른 일 관점에 따르면, 갈파래, 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상 조류의 분말 10 내지 20 중량부, 지충이(Sargassum thunbergii) 10 내지 20 중량부, 대두박(soybean meal) 5 내지 10 중량부, 조개분말(shell fish powder) 5 내지 10 중량부, 패각분말(Shell powder) 1 내지 3 중량부, 인산칼슘(calcium phosphate) 1 내지 3 중량부, 효모 추출물(yeast extract) 3 내지 8 중량부, 콩 레시틴(soyabean lecithin) 2 내지 6 중량부, 미네랄(mineral) 0.2 내지 0.7 중량부, 비타민(vitamin) 0.2 내지 0.7 중량부, 및 바다 진흙(sea mud) 20 내지 60 중량부를 혼합하여 혼합물을 제조하는 단계; 및 상기 혼합물을 건조시켜 고형화하는 단계를 포함하는 천식 개선 효과가 강화된 해삼 양식용 사료조성물의 제조방법이 제공된다. According to another aspect of the invention, 10 to 20 parts by weight of one or two or more algae selected from the group consisting of brown seaweed, kelp and Nannochloropsi s sp., Sargassum thunbergii 10 To 20 parts by weight, 5 to 10 parts by weight of soybean meal, 5 to 10 parts by weight of shell fish powder, 1 to 3 parts by weight of shell powder, 1 to 3 of calcium phosphate Parts by weight, yeast extract 3 to 8 parts by weight, soyabean lecithin 2 to 6 parts by weight, minerals 0.2 to 0.7 parts by weight, vitamins 0.2 to 0.7 parts by weight, and sea mud (sea mud) mixing 20 to 60 parts by weight to prepare a mixture; And there is provided a method for producing a sea cucumber culture feed composition is enhanced asthma improving effect comprising the step of drying the mixture to solidify.

상기한 바와 같이 본 발명의 천식 개선물질을 포함하는 해삼사료 조성물 및 그 제조방법은 당지질 포함량이 높은 갈조류분을 주원료로 사용하여 자연산 해삼에 비해서 천식개선 효능이 낮거나 거의 없는 것으로 알려진 양식해삼에 급여하여 천식 개선물질을 다량 포함하는 해삼을 생산하는 효과를 구현할 수 있다. 물론 이러한 효과에 의해 본 발명의 범위가 한정되는 것은 아니다. As described above, the sea cucumber feed composition comprising the asthma improving material of the present invention and a method for preparing the same are fed to cultured sea cucumber, which is known to have low or almost no asthma improvement efficacy compared to wild sea cucumber, using brown algae containing a high content of glycolipid as a main ingredient. It can achieve the effect of producing sea cucumber containing a large amount of asthma improving material. Of course, the scope of the present invention is not limited by these effects.

도 1은 본 발명의 실험 사료를 이용한 해삼 사육실험을 위해 설계된 순환여과시스템(RAS)의 개요도이다.
도 2는 본 발명의 실험 사료를 이용한 해삼 사육실험을 위해 설계된 순환여과시스템(RAS)의 사진이다.
도 3은 본 발명의 실험 사료를 급여한 해삼의 비성장속도(SGR)를 분석한 그래프이다. 알파벳 소문자는 같은 처리군 간에 유의한 차이 (P <0.05)를 나타내며 막대는 표준 오차를 나타낸다.
도 4는 본 발명의 실험 사료를 급여한 해삼의 섭취율(IR)을 분석한 그래프이다. 알파벳 소문자는 같은 처리군 간에 유의한 차이 (P <0.05)를 나타내며 막대는 표준 오차를 나타낸다.
도 5는 본 발명의 실험 사료를 급여한 해삼의 배설물 생산율(FPR)을 분석한 그래프이다. 알파벳 소문자는 같은 처리군 간에 유의한 차이 (P <0.05)를 나타내며 막대는 표준 오차를 나타낸다.
도 6은 본 발명의 실험 사료를 급여한 해삼의 식이 전환 효율(FCE)을 분석한 그래프이다. 알파벳 소문자는 같은 처리군 간에 유의한 차이 (P <0.05)를 나타내며 막대는 표준 오차를 나타낸다.
도 7은 본 발명의 실험 사료를 급여한 해삼의 추출물의 투여 후 알레르기성 호흡기 질환 유도 마우스 모델 제조과정을 나타내는 사진이다.
도 8은 본 발명의 실험 사료를 급여한 해삼의 추출물을 마우스 모델에 투여에 따른 항 염증성 사이토카인인 IL-10 유전자 발현 수준을 분석한 그래프이다. 알파벳 소문자는 같은 처리군 간에 유의한 차이(P <0.05)를 나타내며 막대는 표준 오차를 나타낸다.1 is a schematic diagram of a circulating filtration system (RAS) designed for the sea cucumber breeding experiment using the experimental feed of the present invention.
Figure 2 is a photograph of a circulating filtration system (RAS) designed for the sea cucumber breeding experiment using the experimental feed of the present invention.
Figure 3 is a graph analyzing the specific growth rate (SGR) of sea cucumbers fed the experimental feed of the present invention. Lowercase letters represent significant differences (P <0.05) between the same treatment groups and bars represent standard error.
Figure 4 is a graph analyzing the intake rate (IR) of sea cucumbers fed the experimental feed of the present invention. Lowercase letters represent significant differences (P <0.05) between the same treatment groups and bars represent standard error.
5 is a graph analyzing the fecal production rate (FPR) of sea cucumbers fed the experimental feed of the present invention. Lowercase letters represent significant differences (P <0.05) between the same treatment groups and bars represent standard error.
Figure 6 is a graph analyzing the dietary conversion efficiency (FCE) of sea cucumbers fed the experimental feed of the present invention. Lowercase letters represent significant differences (P <0.05) between the same treatment groups and bars represent standard error.
Figure 7 is a photograph showing the process of manufacturing an allergic respiratory disease-induced mouse model after administration of the extract of sea cucumber fed the experimental feed of the present invention.
Figure 8 is a graph analyzing the level of IL-10 gene expression of anti-inflammatory cytokines according to administration of the extract of sea cucumbers fed the experimental feed of the present invention to the mouse model. Lowercase letters represent significant differences (P <0.05) between the same treatment groups and bars represent standard error.

용어의 정의:Definition of Terms:

본 문서에서 사용되는 용어 "해삼(Apostichopus japonicus)"은 돌기해삼이라고 불리고 있으며 생물학적 분류체계에 따르면 극피동물문(Echinodermata), 해삼강(Holothuroidea), 해삼과(Stichopodiae) 여자삼속(Apostichopus)에 속한다. 건강 기능성물질인 사포닌과 콘드로이친을 다량 포함하고 있으므로 예로부터 바다의 인삼이라 부른다. The term "sea cucumber ( Apostichopus japonicus )" is used in this document and is called perilla sea cucumber, and according to the biological classification system, it belongs to the Echinodermata, the sea cucumber river (Holothuroidea), the sea cucumber family (Stichopodiae) and the Apostichopus . Since it contains large amounts of saponin and chondroitin, which are health functional substances, it is called sea ginseng since ancient times.

본 문서에서 사용되는 용어 "Interleukin 10(IL-10)"은 인간 사이토카인 합성 억제 인자(CSIF)로도 알려져 있으며 항 염증성 사이토카인으로 IL-10 유전자에 의해 암호화된다. IL-10은 2개의 IL-10 수용체-1과 2개의 IL-10 수용체-2 단백질로 구성된 수용체 복합체를 통해 신호를 보낸다. The term "Interleukin 10 (IL-10)" as used herein is also known as human cytokine synthesis inhibitory factor (CSIF) and is an anti-inflammatory cytokine encoded by the IL-10 gene. IL-10 signals through a receptor complex consisting of two IL-10 receptor-1 and two IL-10 receptor-2 proteins.

발명의 상세한 설명:Detailed description of the invention:

본 발명의 일 관점에 따르면, 갈파래, 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상 조류의 분말 10 내지 20 중량부, 지충이(Sargassum thunbergii) 10 내지 20 중량부, 대두박(soybean meal) 5 내지 10 중량부, 조개분말(shell fish powder) 5 내지 10 중량부, 패각분말(Shell powder) 1 내지 3 중량부, 인산칼슘(calcium phosphate) 1 내지 3 중량부, 효모 추출물(yeast extract) 3 내지 8 중량부, 콩 레시틴(soyabean lecithin) 2 내지 6 중량부, 미네랄(mineral) 0.2 내지 0.7 중량부, 비타민(vitamin) 0.2 내지 0.7 중량부, 및 바다 진흙(sea mud) 20 내지 60 중량부를 포함하는, 천식 개선 효과가 강화된 해삼의 생산을 위한 해삼 양식용 사료조성물이 제공된다. According to an aspect of the present invention, 10 to 20 parts by weight of one or two or more algae selected from the group consisting of brown seaweed, kelp and Nanclochloros s sp., 10 to 20 parts by weight, Sargassum thunbergii 20 parts by weight, 5-10 parts by weight of soybean meal, 5-10 parts by weight of shell fish powder, 1-3 parts by weight of shell powder, 1-3 parts by weight of calcium phosphate Parts, 3 to 8 parts by weight of yeast extract, 2 to 6 parts by weight of soyabean lecithin, 0.2 to 0.7 parts by weight mineral, 0.2 to 0.7 parts by weight vitamin, and sea mud ( Sea mud) Provided with a sea cucumber culture feed composition for the production of sea cucumber enhanced asthma-improving effect, including 20 to 60 parts by weight.

상기 해삼 양식용 사료조성물은 더 바람직하게는 갈파래, 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상 조류의 분말 13 내지 17 중량부, 지충이 13 내지 17 중량부, 대두박 6 내지 9 중량부, 조개분말 6 내지 9 중량부, 패각 분말 1.5 내지 2.5 중량부, 인산칼슘 1.5 내지 2.5 중량부, 효모추출물 4 내지 7 중량부, 콩 레시틴 3 내지 5 중량부, 미네랄 0.3 내지 0.6 중량부, 비타민 0.3 내지 0.6 중량부, 및 바다 진흙 30 내지 50 중량부를 포함할 수 있다.The sea cucumber aquaculture feed composition is more preferably 13 to 17 parts by weight of one or two or more algae powder selected from the group consisting of brown seaweed, kelp and Nannochloropsi s sp. Parts by weight, soybean meal 6 to 9 parts by weight, shell powder 6 to 9 parts by weight, shell powder 1.5 to 2.5 parts by weight, calcium phosphate 1.5 to 2.5 parts by weight, yeast extract 4 to 7 parts by weight, soy lecithin 3 to 5 parts by weight, 0.3 to 0.6 parts by weight of minerals, 0.3 to 0.6 parts by weight of vitamins, and 30 to 50 parts by weight of sea mud.

상기 해삼 양식용 사료조성물은 가장 바람직하게는 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상 조류의 분말 14 내지 16 중량%, 지충이 14 내지 16 중량%, 대두박 7 내지 9 중량%, 조개분말 7 내지 9 중량%, 패각 분말 1.5 내지 2.5 중량%, 인산칼슘 1.5 내지 2.5 중량%, 효모추출물 4 내지 6 중량%, 콩 레시틴 3 내지 5 중량%, 미네랄 0.4 내지 0.6 중량%, 비타민 0.4 내지 0.6 중량%, 및 잔부의 바다 진흙으로 구성될 수 있다.The sea cucumber aquaculture feed composition is most preferably 14 to 16% by weight of powder of one or more algae selected from the group consisting of kelp and Nannochloropsi s sp., 14 to 16% by weight of worms , Soybean meal 7 to 9% by weight, clam powder 7 to 9% by weight, shell powder 1.5 to 2.5% by weight, calcium phosphate 1.5 to 2.5% by weight, yeast extract 4 to 6% by weight, soy lecithin 3 to 5% by weight, mineral 0.4 To 0.6% by weight, 0.4 to 0.6% by weight of vitamins, and the balance of sea mud.

본 발명의 다른 일 관점에 따르면, 갈파래, 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상 조류의 분말 10 내지 20 중량부, 지충이(Sargassum thunbergii) 10 내지 20 중량부, 대두박(soybean meal) 5 내지 10 중량부, 조개분말(shell fish powder) 5 내지 10 중량부, 패각분말(Shell powder) 1 내지 3 중량부, 인산칼슘(calcium phosphate) 1 내지 3 중량부, 효모 추출물(yeast extract) 3 내지 8 중량부, 콩 레시틴(soyabean lecithin) 2 내지 6 중량부, 미네랄(mineral) 0.2 내지 0.7 중량부, 비타민(vitamin) 0.2 내지 0.7 중량부, 및 바다 진흙(sea mud) 20 내지 60 중량부를 혼합하여 혼합물을 제조하는 단계; 및 상기 혼합물을 건조시켜 고형화하는 단계를 포함하는 천식 개선 효과가 강화된 해삼 양식용 사료조성물의 제조방법이 제공된다. According to another aspect of the invention, 10 to 20 parts by weight of one or two or more algae selected from the group consisting of brown seaweed, kelp and Nannochloropsi s sp., Sargassum thunbergii 10 To 20 parts by weight, 5 to 10 parts by weight of soybean meal, 5 to 10 parts by weight of shell fish powder, 1 to 3 parts by weight of shell powder, 1 to 3 of calcium phosphate Parts by weight, yeast extract 3 to 8 parts by weight, soyabean lecithin 2 to 6 parts by weight, minerals 0.2 to 0.7 parts by weight, vitamins 0.2 to 0.7 parts by weight, and sea mud (sea mud) mixing 20 to 60 parts by weight to prepare a mixture; And there is provided a method for producing a sea cucumber culture feed composition is enhanced asthma improving effect comprising the step of drying the mixture to solidify.

대부분의 해삼은 침전물을 유기 물질로 섭취하는 퇴적물 섭식자(deposit feeders)이다. 전 세계적으로 해삼양식 생산량이 증가함에 따라 해삼양식은 배합사료 급여에 의한 양적증가에만 치중되어 있으며, 건강식품으로의 질적인 측면에서의 생산이 무시되고 있다. 또한 양식해삼은 자연산 해삼에 비해서 천식개선 효능이 낮거나 거의 없는 것으로 나타나고 있다. 이에 본 발명자들은 양식 해삼에서 천식을 개선할 수 있는 물질을 포함한 해삼을 생산하기 위해 예의노력한 결과 기존 상용의 해조분말 지충이 대신에 당지질 포함량이 높은 갈조류분(미역, 다시마) 또는 오메가-3 지방산(EPA)을 포함하는 해수 클로렐라(Nannochloropsis)를 주원료로서 제공함으로써 종래 해삼 양식방법과 비교하여 성장이 우수하고 천식 개선물질을 다량 포함하는 해삼을 생산하기 위한 사료조성물 및 그 제조방법을 개발하여 본 발명을 완성하였다. Most sea cucumbers are deposit feeders that consume sediment as organic material. As sea cucumber culture production increases worldwide, sea cucumber culture is focused only on the quantitative increase by the feeding of formulated feed, and the production in terms of quality as a health food is ignored. In addition, cultured sea cucumber has been shown to have low or almost no asthma efficacy compared to wild sea cucumber. Therefore, the present inventors have diligently tried to produce sea cucumbers containing substances that can improve asthma in cultured sea cucumbers, and instead of conventional seaweed powder worms, brown algae powder (seaweed, kelp) or omega-3 fatty acids (high seaweed) By providing a seawater chlorella ( Nanochloropsis ) containing EPA) as a main raw material to develop a feed composition and production method for producing a sea cucumber which is excellent in growth compared to the conventional sea cucumber culture method and contains a large amount of asthma-improving material and the present invention Completed.

이하, 실시예를 통하여 본 발명을 더 상세히 설명한다. 그러나 본 발명은 이하에서 개시되는 실시예에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 수 있는 것으로, 이하의 실시예는 본 발명의 개시가 완전하도록 하며, 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. However, the present invention is not limited to the embodiments disclosed below, but can be implemented in various forms, and the following embodiments are intended to complete the disclosure of the present invention, the scope of the invention to those skilled in the art It is provided to inform you completely.

실시예 1: 실험 사료(feed stuff)Example 1: Experimental Feed Stuff

본 발명자들은 천식 개선물질을 포함하는 해삼사료 조성물을 제조를 위해 기본사료 또는 천식개선 후보물질을 포함하는 총 6종의 실험 사료를 제조하였다. 구체적으로 기본사료 100%로 제조한 사료는 대조군으로 사용하였고(control) 기본사료 85%에 갈파래분(Ulva lactuca powder) 15%를 첨가한 실험군(Diet 1), 기본사료 85%에 미역분(Undaria pinnatifida powder) 15%를 첨가한 실험군(Diet 2), 기본사료 85%에 다시마분(Laminaria japonica powder) 15%를 첨가한 실험군(Diet 3), 기본사료 85%에 미세조류분(Schizochytrium powder) 15%를 첨가한 실험군(Diet 4), 및 기본사료 85%에 해수클로렐라분(Nannochloropsis powder) 15%를 첨가한 실험군(Diet 5)을 제조하였다. 상기 실험군의 해조류 중 갈파래, 미역, 다시마는 시중에 판매되고 있는 건조된 원료를 구입하여 200-μm mesh의 미세 분말로 갈아서 사료에 첨가하였고 시조키트리움과 해수클로렐라는 양식용 사료 판매처로부터 구입하여 동결건조과정을 거쳐 분말화 후 -20℃ 냉동 보관하여 사용하였으며 경상대학교의 해양생물 및 양식학 실험실에서 9주 동안 상기 제조한 실험 사료를 이용하여 실험동물 및 사료 성능 실험을 수행하였다The present inventors prepared a total of six kinds of experimental feed including a basic feed or an asthma improvement candidate for preparing a sea cucumber feed composition comprising an asthma improving material. Specifically, the feed prepared with 100% of the basic feed was used as a control (control) and the experimental group (Diet 1) to which 15% of Ulva lactuca powder was added to 85% of the basic feed, and the seaweed powder ( Undaria ) to 85% of the basic feed. Experiment group added 15% pinnatifida powder (Diet 2), experiment group added 15% kelp powder ( Laminaria japonica powder) to 85% basic feed, and Schizochytrium powder 15% to 85% basic feed. An experimental group (Diet 4) to which% was added, and an experimental group (Diet 5) to which 15% of seawater chlorella powder ( Nanochloropsis powder) was added to 85% of the basic feed were prepared. Brown seaweed, seaweed and kelp among the seaweeds of the experimental group purchased dried raw materials commercially available, were ground into fine powder of 200-μm mesh and added to the feed, and Shijo Kitrium and seawater chlorella were purchased from aquaculture feed stores and frozen. After drying, it was powdered and stored at -20 ℃ for freezing. Experimental animal and feed performance experiments were performed using the experimental feed prepared for 9 weeks in the marine biology and aquaculture laboratory of Gyeongsang National University.

기본사료 또는 천식개선 후보물질을 포함하는 총 6종의 실험 사료의 조성Composition of a total of six experimental feeds containing basic feed or candidates for improving asthma 성분ingredient 대조군Control Diet 1Diet 1 Diet 2Diet 2 Diet 3Diet 3 Diet 4Diet 4 Diet 5Diet 5 Ulva lactuca powder Ulva lactuca powder 00 1515 00 00 00 00 Undaria pinnatifida powder Undaria pinnatifida powder 00 00 1515 00 00 00 Laminaria japonica powder Laminaria japonica powder 00 00 00 1515 00 00 Schizochytrium powder Schizochytrium powder 00 00 00 00 1515 00 Nannochloropsis powder Nannochloropsis powder 00 00 00 00 00 1515 Wheat flourWheat flour 1515 00 00 00 00 00 기본 성분Basic ingredients Sargassum thunbergii powder Sargassum thunbergii powder 1515 1515 1515 1515 1515 1515 Soybean mealSoybean meal 88 88 88 88 88 88 Shellfish powderShellfish powder 88 88 88 88 88 88 Shell powderShell powder 22 22 22 22 22 22 Calcium phosphateCalcium phosphate 22 22 22 22 22 22 Yeast extractYeast extract 55 55 55 55 55 55 Soyabean lecithinSoyabean lecithin 44 44 44 44 44 44 MineralMineral 0.50.5 0.50.5 0.50.5 0.50.5 0.50.5 0.50.5 VitaminVitamin 0.50.5 0.50.5 0.50.5 0.50.5 0.50.5 0.50.5 Sea mudSea mud 4040 4040 4040 4040 4040 4040

실시예 2: 일반성분 분석Example 2: General Component Analysis

본 발명자들은 상기에서 제조된 비교예 및 실시예 1 내지 5의 해삼사료에 대해 일반성분 분석을 수행하였다. 구체적으로, 일반성분 분석은 AOAC(Association of Official Analytical Chemists. Washington DC., AOAC. Official method of analysis. 16th ed. U.S.A., 1-43, 1995)방법, 조단백질은 Kjeldahl법으로 분석하였으며, 조지방은 클로로포름(choloroform)과 메탄올(methanol)을 2:1 비율로 혼합한 용액을 용매로 한 Bligh and Dyer추출법(1959)에 준하였다. 수분은 상압가열 건조법으로 105℃의 드라이오븐(dry oven)에서 6시간 동안 건조 후 측정하였고, 조회분은 직접회화법(Dry ashing)으로 600℃ 회화로에서 4시간 동안 태운 후 정량하였다.The inventors performed a general component analysis for the sea cucumber feed of Comparative Examples and Examples 1 to 5 prepared above. Specifically, general component analysis was performed by Association of Official Analytical Chemists.Washington DC., AOAC.Official method of analysis.16th ed.USA., 1-43, 1995), crude protein was analyzed by Kjeldahl method. The solution of chloroform (choloroform) and methanol (methanol) in a 2: 1 ratio was used for Bligh and Dyer extraction (1959). Moisture was measured after drying for 6 hours in a 105 ℃ dry oven (dry oven) by atmospheric pressure drying method, the ash content was quantified after burning for 4 hours in 600 ℃ incinerator by dry ashing (Dry ashing).

그 결과, 대조군의 경우 수분을 제외한 조단백질은 17.7%, 지질 3.4%, 조회분 41.2%로 나타났고, Diet 1의 해삼사료는 조단백질 17%, 조지질 3.7%, 조회분 44.8%로 나타났으며, Diet 2의 해삼사료는 조단백질 17.5%, 조지질 3.1%, 조회분 45.5%로 나타났다. 또한, Diet 3의 해삼사료는 조단백질 17.1%, 조지질 3.1%, 조회분 45.8%로 실험군 중 가장 높은 조회분 함량을 나타내었고 Diet 4의 해삼사료는 조단백질 17.6%, 조지질 7.4%, 조회분 41.5%로 실험군 중 가장 높은 조지질 함량을 나타내었으며 Diet 5의 해삼사료은 조단백질 19.6%, 조지질 5.6%, 조회분 42.1%로 실험군 중 가장 높은 조단백질 함량을 나타내었다. 상기 일반성분 분석 결과를 하기 표 2에 요약하였다. In the control group, crude protein except water was 17.7%, lipid 3.4%, crude ash 41.2%, and sea cucumber feed of Diet 1 was crude protein 17%, crude lipid 3.7%, crude ash 44.8%. Sea cucumber feed of Diet 2 was 17.5% of crude protein, 3.1% of crude lipid, and 45.5% of crude ash. In addition, the sea cucumber feed of Diet 3 showed the highest crude ash content of the experimental group with crude protein of 17.1%, crude lipid of 3.1%, and crude ash of 45.8%. The highest crude protein content of the experimental group was shown in%, and the sea cucumber feed of Diet 5 showed the highest crude protein content of crude protein of 19.6%, crude lipid of 5.6%, crude ash of 42.1%. The general component analysis results are summarized in Table 2 below.

총 6종의 실험 사료의 일반성분 분석Analysis of General Components of Six Experimental Feeds 일반 조성(%)General composition (%) 대조군Control Diet 1Diet 1 Diet 2Diet 2 Diet 3Diet 3 Diet 4Diet 4 Diet 5Diet 5 조단백질Crude protein 17.7417.74 17.0317.03 17.4817.48 16.8716.87 17.5917.59 19.6419.64 조지질Geography 3.393.39 3.723.72 3.113.11 3.323.32 7.447.44 5.645.64 조회분View minutes 41.2041.20 44.8044.80 45.5545.55 45.8545.85 41.541.5 42.1042.10

실시예 3: 해삼 사육실험 Example 3: Sea cucumber breeding experiment

본 발명자들은 상기 실시예 1에서 제조한 총 6종의 실험사료를 이용하여 해삼 사육실험을 수행하였다. 구체적으로, 본 발명에 사용한 돌기해삼(Apostichopus japonicus)은 평균체중 3.0g으로 1,000마리를 경남 통영시 산양면의 종묘생산업체에서 분양받아 실험실로 운반하여 1,000 L 수조에 입식한 후, 실험환경에 적응할 수 있도록 17℃, 32 psu의 해수에서 기초사료를 공급하면서 5주간 예비사육을 실시하였고 죽은 개체는 폐기처분한 후, 평균체중 3.0±0.1 g인 건강한 해삼 480마리를 선발하여 96 L의 4각 수조에 각각 20개체씩 수용하고 해삼사육을 위하여 설계한 순환여과시스템(RAS)에서 각 실험군당 사육실험을 4회 반복하였다(수조당 20마리씩x 4반복 x6개 사료구 = 480마리). 각 실험수조의 사육수 순환율은 20 주기/일로 조절하였고 실험기간동안의 사육환경은 수온 18℃, 염분 32 psu, pH 7.7~8.3, 용존산소 5.0~7.0 mg/L 및 총 암모니아 0.25ppm 이하의 범위로 유지하였으며 1일 사료공급량은 해삼체중의 5%를 1일 1회(17:00) 충분하게 공급하면서 9주간 사육을 실시하였다(도 1 및 2). The present inventors carried out sea cucumber breeding experiment using a total of six experimental feeds prepared in Example 1. Specifically, Apostichopus japonicus used in the present invention has an average weight of 3.0g and is sold at a seedling producer of Sanyang-myeon, Tongyeong-si, Gyeongnam, and transported to a laboratory for stocking in a 1,000 L tank, so that it can adapt to the experimental environment. Pre-breeding was carried out for 5 weeks while feeding the basic feed in 17 ℃ and 32 psu of seawater. After discarding the dead individuals, 480 healthy sea cucumbers with an average weight of 3.0 ± 0.1 g were selected and placed in a 96-L quadrangular tank. In a circular filtration system (RAS) designed to accommodate 20 animals and breeding sea cucumbers, four experiments were repeated for each experimental group (20 animals per tank x 4 repetitions x 6 feeds = 480 animals). The breeding water circulation rate of each experimental tank was adjusted to 20 cycles / day, and the breeding environment during the experiment period was 18 ℃, salinity 32 psu, pH 7.7 ~ 8.3, dissolved oxygen 5.0 ~ 7.0 mg / L and 0.25ppm total ammonia. The daily feed was maintained for 9 weeks while feeding enough 5% of the sea cucumber body weight once a day (17:00) (Figs. 1 and 2).

실시예 4: 샘플 수집 Example 4: Sample Collection

본 발명자들은 실험을 시작하기 전에 초기 샘플로 24마리의 해삼을 수집하였고 실험 기간 동안 해삼에게 1일 1회(약 1700 시간) 사료를 급여하였다. 먹지 않는 사료(uneaten feed)는 24시간 후에 사이펀(siphon)으로 수집하였고 65℃에서 일정한 무게로 건조하였다. 해삼 배설물도 1일 1회(1600 시간) 사이펀으로 수집하였고 상기 배설물을 65℃에서 일정한 무게로 건조시킨 후 추가 분석을 위해 각 수조의 배설물을 수거하였다. 9주간의 실험이 끝날 때 모든 실험용 해삼은 내장 제거를 위해 2일 동안 사료 급여를 중단하였고 무게를 측정 한 다음 일정한 무게가 될 때까지 65℃에서 건조하였다.We collected 24 sea cucumbers as initial samples before starting the experiment and fed sea cucumbers once daily (about 1700 hours) during the experiment. Uneated feed was collected with siphon after 24 hours and dried to constant weight at 65 ° C. Sea cucumber droppings were also collected by siphon once a day (1600 hours) and the droppings were dried to a constant weight at 65 ° C. and then the droppings from each tank were collected for further analysis. At the end of the 9-week experiment, all experimental sea cucumbers were stopped feeding for 2 days to remove viscera, weighed and dried at 65 ° C until constant weight.

실시예 5: 데이터 계산 Example 5: Data Calculation

본 발명의 일 실시예에 따라 상기 제조한 해삼사료의 급여에 따른 실험군의 생존율(survival rate, SR), 비성장속도(specific growth rate, SGR), 섭취율(ingestion rate, IR), 배설물 생산율(feces production rate, FPR), 및 식이 전환 효율(food conversion efficiency, FCE)은 하기 공식으로 계산하였다. Survival rate (SR), specific growth rate (SGR), intake rate (IR), feces production rate (feces) of the experimental group according to the feed of the prepared sea cucumber feed according to an embodiment of the present invention production rate (FPR), and food conversion efficiency (FCE) were calculated by the following formula.

생존률(SR):Survival Rate (SR):

Figure pat00001
Figure pat00001

상기 식에서 N2는 최종 생존개체수, N1은 최초 개체수를 나타낸다.In the above formula, N2 represents the final number of surviving individuals and N1 represents the initial population.

비성장속도(SGR):Specific Growth Rate (SGR):

Figure pat00002
Figure pat00002

상기 식에서 W 1 은 최초무게, W 2 는 최종무게, I는 섭식한 사료량 및 T는 사육기간을 나타낸다. Where W 1 is the initial weight, W 2 is the final weight, I is the feeding amount and T is the breeding period.

섭취율(IR):Intake Rate (IR):

Figure pat00003
Figure pat00003

상기 식에서 I는 섭취한 총 사료의 건조 중량을 나타낸다. Where I represents the dry weight of the total feed consumed.

배설물 생산율(FPR):Excretion Production Rate (FPR):

Figure pat00004
Figure pat00004

상기 식에서 F는 배설물의 건조 중량을 나타낸다. In the above formula, F represents the dry weight of feces.

식이 전환 효율(FCE):Dietary Conversion Efficiency (FCE):

Figure pat00005
Figure pat00005

상기 식에서 W 1 은 최초무게, W 2 는 최종무게를 나타낸다. In the above formula, W 1 represents the initial weight and W 2 represents the final weight.

모든 통계분석은 SPSS 16.0 프로그램을 사용하여 One-way ANOVA test를 실시한 후 Duncan smultiple rang test(Duncan, 1995)로 평균 간의 유의성(P<0.05)을 검정하였다.All statistical analyzes were performed using the SPSS 16.0 program and the one-way ANOVA test followed by Duncan smultiple rang test (Duncan, 1995) to test the significance between the means (P <0.05).

실시예 6: 해삼 추출물의 준비 Example 6: Preparation of Sea Cucumber Extract

해삼 추출물을 제조하기 위해 상기 실험 해삼을 세척 후 내장 기관을 제거하였고 작은 조각으로 자르고 균질화하였다. 그 후 시료 150 g을 증류수 300 ml에서 20분간 끓여 주었다. 물에서 고체 물질을 제거한 후, 혼합물이 50% 감소될 때까지 전자레인지(microwave)를 사용하여 상기 끓인 물을 증발시켰고 추출물을 500 x g에서 10분간 원심분리한 후 5배량의 100% 에틸 알코올을 상등액에 첨가하였으며 20 ℃에서 24시간 배양한 후 상기 상등액을 제거하였다. 상기 추출 펠렛을 70% 에틸 알코올로 세척하였고 상기와 동일한 조건으로 원심분리하였으며 상등액을 제거하고 펠릿을 진공하에 증발시킨 후 최종 추출물은 20 ml의 증류수에 펠릿을 재현탁시켜 제조하였다(Lee et al., Am J Chin Med. 44:1663-74. 2016).To prepare the sea cucumber extract, the experimental sea cucumber was washed, the internal organs were removed, cut into small pieces, and homogenized. Thereafter, 150 g of the sample was boiled in 300 ml of distilled water for 20 minutes. After removing the solids from the water, the boiled water was evaporated using a microwave until the mixture was reduced by 50%, the extract was centrifuged at 500 xg for 10 minutes and then 5 times 100% ethyl alcohol supernatant. The supernatant was removed after incubation at 20 ° C. for 24 hours. The extracted pellet was washed with 70% ethyl alcohol, centrifuged under the same conditions as above, the supernatant was removed and the pellet was evaporated under vacuum and the final extract was prepared by resuspending the pellet in 20 ml of distilled water (Lee et al. , Am J Chin Med. 44: 1663-74. 2016).

실시예 7: IL-10 유전자 발현 분석 Example 7: IL-10 Gene Expression Analysis

본 발명자들은 IL-10 유전자 발현을 분석하기 위해, 실험용 사료-급여 해삼 추출물 10 ㎍/㎖를 이용하여 2시간 동안 마우스의 비장 세포(splenocyte)에 자극하였다. 그 후 총 RNA를 제조사의 지침에 따라 Qiazol 시약(Qiagen Science, USA)으로 분리하였고 제조사의 지침에 따라, 총 RNA의 2 ㎍을 M-MLV 역전사 효소(Promega, USA)를 사용하여 전사시켰고 IL-10 mRNA 발현 수준은 iCyclerTM(Bio-Rad Laboratories, Hercules, CA, USA)을 사용하여 실시간 PCR로 합성하였다. 또한 GAPDH를 참조 유전자(reference gene)로 사용하였다. 상기 실시간 PCR로에 사용한 프라이머 정보를 하기 표 3에 요약하였다. To analyze IL-10 gene expression, the inventors stimulated splenocytes of mice for 2 hours using 10 μg / ml of experimental feed-sealed sea cucumber extract. The total RNA was then separated with Qiazol reagent (Qiagen Science, USA) according to the manufacturer's instructions, and according to the manufacturer's instructions, 2 μg of total RNA was transcribed using M-MLV reverse transcriptase (Promega, USA) and IL- 10 mRNA expression levels were synthesized by real time PCR using iCycler ™ (Bio-Rad Laboratories, Hercules, CA, USA). GAPDH was also used as a reference gene. Primer information used in the real time PCR furnace is summarized in Table 3 below.

IL-10 및 GAPDH 프라이머 서열IL-10 and GAPDH Primer Sequences 프라이머primer 염기서열(5'-->3')Sequence (5 '-> 3') 서열번호SEQ ID NO: GAPDH FGAPDH F TAC CCC CAA TGT GTC CGT CTAC CCC CAA TGT GTC CGT C 1One GAPDH RGAPDH R AAG AGT GGG AGT TGC TGT TGA AGAAG AGT GGG AGT TGC TGT TGA AG 22 IL-10 FIL-10 F GCT ATG CTG CCT GGT CTT ACT GGCT ATG CTG CCT GGT CTT ACT G 33 IL-10 RIL-10 R TCC AGC TGG TCC TTT GTT TGTCC AGC TGG TCC TTT GTT TG 44

실험예 1: 생존과 성장 분석 Experimental Example 1: Survival and Growth Analysis

본 발명의 일 실시예에 따라 제조한 총 6종의 사료를 급여한 후 상기 사료의 성능을 분석하였다. 본 발명의 모든 해삼은 9주간의 급여 기간이 끝날 때까지 살아 있었고 해삼의 생장 능력은 사료의 종류의 따라 크게 차이가 났다. 해삼의 최종 건습 중량은 Diet 1 실험군에서 가장 높았고 대조군에서 가장 낮게 나타났다(표 4 참조)(P <0.05). After feeding a total of six feeds prepared according to an embodiment of the present invention, the performance of the feed was analyzed. All of the sea cucumbers of the present invention were alive until the end of the nine-week pay period, and the growth ability of the sea cucumbers varied greatly depending on the type of feed. The final wet and dry weight of sea cucumber was highest in the Diet 1 experimental group and lowest in the control group (see Table 4) (P <0.05).

또한, 상기 Diet 1 사료를 급여한 해삼에서 가장 높은 비성장속도(SGR, 1.58 % d-1)를 관찰하였으나 대조군은 Diet 1 및 Diet 3 보다 유의적으로 낮게 나타났고(P <0.05) Diet 5는 Diet 4와 비교하여 유의한 차이를 나타내지 않았다(P> 0.05)(도 3). 상기 해삼에 급여한 사료에 따른 습식 및 건식 중량을 하기 표 4에 요약하였다. In addition, the highest growth rate (SGR, 1.58% d -1 ) was observed in the sea cucumbers fed the Diet 1 diet, but the control group was significantly lower than Diet 1 and Diet 3 (P <0.05). There was no significant difference compared to Diet 4 (P> 0.05) (FIG. 3). Wet and dry weight according to the feed fed to the sea cucumber is summarized in Table 4 below.

해삼사료의 급여에 따른 성능분석 결과 Performance Analysis Results According to Salary of Sea Cucumber Feed 항목Item 초기
Wet Weight(g)Early
Wet Weight (g) 초기
Dry Weight(g)Early
Dry Weight (g) 최종
Wet Weight(g)final
Wet Weight (g) 최종
Dry Weight(g)final
Dry Weight (g) 대조군Control 3.04±0.093.04 ± 0.09 0.280.28 5.07±0.925.07 ± 0.92 0.470.47 Diet 1Diet 1 2.93±0.122.93 ± 0.12 0.270.27 7.65±0.857.65 ± 0.85 0.710.71 Diet 2Diet 2 2.94±0.112.94 ± 0.11 0.270.27 5.94±0.625.94 ± 0.62 0.550.55 Diet 3Diet 3 3.03±0.073.03 ± 0.07 0.280.28 7.27±0.457.27 ± 0.45 0.670.67 Diet 4Diet 4 2.92±0.102.92 ± 0.10 0.270.27 6.16±0.156.16 ± 0.15 0.570.57 Diet 5Diet 5 3.02±0.063.02 ± 0.06 0.280.28 7.05±0.187.05 ± 0.18 0.650.65

실험예 2: 섭취율(IR), 배설물 생산율(FPR) 및 식이 전환 효율(FCE)Experimental Example 2: Intake rate (IR), fecal production rate (FPR) and dietary conversion efficiency (FCE)

본 발명자들은 실험 사료를 해삼에게 급여 후 섭취율, 배설물 생산율 및 식이 전환 효율을 분석하였다. 실험군 중에서 Diet 3 및 Diet 1는 다른 사료를 급여한 해삼보다 유의하게 높은 섭취율(각각 0.62 및 0.59 g g-1 d-1) 및 배설물 생산율(각각 0.53 및 0.52 g g-1 d-1)을 나타내었다(P<0.05). 한편 대조군의 해삼은 모든 실험군 중에서 가장 낮은 섭취율(0.32 g g-1 d-1)(P <0.05)과 배설물 생산율(0.26 g g-1 d-1) (P <0.05)을 나타내었다(도 4 및 5). 또한 Diet 2를 급여한 해삼은 가장 낮은 식이 전환 효율을 나타내었으나(2.21%) Diet 5를 급여한 해삼은 다른 실험 사료를 급여한 해삼과 비교하여 현저히 높은 식이 전환 효율(3.74%)을 나타내었다(도 6)(P <0.05). The present inventors analyzed the intake rate, excreta production rate and dietary conversion efficiency after feeding the experimental feed sea cucumber. In the experimental group, Diet 3 and Diet 1 showed significantly higher intake rates (0.62 and 0.59 gg -1 d -1 ) and excreta production rates (0.53 and 0.52 gg -1 d -1 , respectively) than the other sea cucumbers fed the other diets ( P <0.05). Meanwhile, sea cucumbers of the control group showed the lowest intake rate (0.32 gg −1 d −1 ) (P <0.05) and fecal production rate (0.26 gg −1 d −1 ) (P <0.05) among all the experimental groups (FIGS. 4 and 5). ). The sea cucumbers fed Diet 2 had the lowest dietary conversion efficiency (2.21%), but the sea cucumbers fed Diet 5 had a significantly higher dietary conversion efficiency (3.74%) compared to the sea cucumbers fed the other diets. 6) (P <0.05).

실험예 3: IL-10 유전자 발현 수준 Experimental Example 3: IL-10 Gene Expression Levels

본 발명자들은 천식 개선물질을 포함하는 해삼 급여 사료에 적합한 해조류를 선정하기 위해 항 염증성 사이토카인(anti-inflammatory cytokine)인 IL-10 유전자 발현 수준을 분석하였다.The present inventors analyzed the level of IL-10 gene expression, an anti-inflammatory cytokine, in order to select seaweeds suitable for feeding sea cucumber supplements containing asthma improving substances.

구체적으로 알레르기성 호흡기 질환 유도 마우스 모델을 이용하여 천식을 유도하기 전에 본 발명의 실험 사료 6종을 급여한 해삼의 추출물(100 mg/ml)을 마우스당 200 μl씩 2일 간격으로 7번 경구투여 하였고 1일 경과 후, 75 μg 난알부민(ovalbumin, OVA)과 2μg의 백반(alum)을 1일 1회씩 2일 동안 복강 내 주사하였으며 6일 뒤 다시 1일 1회씩 2일 동안 상기와 동량의 난알부민과 백반을 복강 내 주사하였다. 그 후 6일 경과시점에서 50μg의 난알부민을 비강 내 1일 1회씩 2일 동안 투여하였고 6일 뒤 다시 동량의 난알부민을 비강 내 1일 1회씩 2일 동안 투여하여 천식을 유도하였다. 그 후, 천식 유도 여부를 확인하기 위해, 메타콜린(methacholine)을 이용하여 기도 저항성을 측정하였고, 마우스를 희생시킨 후 비장세포(splenocytes)를 적출하여 천식 개선 물질인 IL-10 유전자 발현을 측정하였다(도 7). Specifically, before inducing asthma using an allergic respiratory disease-induced mouse model, the extract of sea cucumber (100 mg / ml) fed six experimental feeds of the present invention was orally administered 7 times at 200 μl per mouse, two days apart. After 1 day, 75 μg of ovalbumin (OVA) and 2 μg of alum were intraperitoneally injected once a day for 2 days, and after 6 days, the same amount of egg for 2 days Albumin and vitiligo were injected intraperitoneally. After 6 days, 50 μg of egg albumin was administered once a day for 2 days in the nasal cavity. After 6 days, the same amount of egg albumin was administered once a day for 2 days to induce asthma. Then, to determine whether asthma induction, airway resistance was measured using methacholine (methacholine), and splenocytes were removed after sacrifice of mice to measure IL-10 gene expression, an asthma improving substance. (FIG. 7).

그 결과, 천식 개선 물질인 IL-10 유전자 발현 수준은 Diet 1, Diet 3와 및 Diet 5 실험군에서 유의하게 증가하였고 Diet 1 즉 갈파래(Ulva lactuca)를 급여하였을 때 가장 높게 나타났다. 그러나 대조군, Diet 2 및 Diet 4 실험군의 경우는 증가하지 않았고 유의한 차이를 나타내지 않았다(도 8). 상기 결과는 본 발명의 해삼 사료인 Diet 1(갈파래), Diet 3(다시마) 및 Diet 5(클로렐라)에 포함된 해조류가 IL-10 유전자 발현을 상향 조절할 수 있음을 시사한다.As a result, IL-10 gene expression level was significantly increased in Diet 1, Diet 3 and Diet 5 experimental group, and was highest when diet 1, Ulva lactuca . However, the control group, Diet 2 and Diet 4 experimental group did not increase and did not show a significant difference (Fig. 8). The above results suggest that seaweeds included in Diet 1 (brown seaweed), Diet 3 (Kashima) and Diet 5 (Chlorella), which are sea cucumber feed of the present invention, may upregulate IL-10 gene expression.

결론적으로 본 발명의 천식 개선물질(IL-10)을 포함하는 해삼사료 조성물 및 그 제조방법은 천식 개선물질을 거의 포함하지 않은 종래 해삼 양식방법과 비교하여 해삼의 성장이 향상되고 천식 개선물질도 다량 포함하고 있으므로 해삼을 비롯한 다양한 수산생물 양식의 급여사료로 활용될 수 있다.In conclusion, the sea cucumber feed composition comprising the asthma improving material (IL-10) of the present invention and its manufacturing method compared with the conventional sea cucumber culture method containing almost no asthma improving material, the growth of sea cucumber is improved and asthma improving material is also a large amount It can be used as a feed for aquatic life, including sea cucumbers.

본 발명은 상술한 실시예를 참고로 설명되었으나 이는 예시적인 것에 불과하며, 당해 기술분야에서 통상의 지식을 가진 자라면 이로부터 다양한 변형 및 균등한 다른 실시예가 가능하다는 점을 이해할 것이다. 따라서 본 발명의 진정한 기술적 보호 범위는 첨부된 특허청구범위의 기술적 사상에 의하여 정해져야 할 것이다.Although the present invention has been described with reference to the above-described embodiments, these are merely exemplary, and it will be understood by those skilled in the art that various modifications and equivalent other embodiments are possible. Therefore, the true technical protection scope of the present invention will be defined by the technical spirit of the appended claims.

<110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION GYEONGSANG NATIONAL UNIVERSITY <120> Composition of feed stuff for sea cucumber containing asthma improving substance and manufacturing method thereof <130> PD18-5660 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> GAPDH F <400> 1 tacccccaat gtgtccgtc 19 <210> 2 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> GAPDH R <400> 2 aagagtggga gttgctgttg aag 23 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> IL-10 F <400> 3 gctatgctgc ctggtcttac tg 22 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-10 R <400> 4 tccagctggt cctttgtttg 20 <110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION GYEONGSANG NATIONAL UNIVERSITY <120> Composition of feed stuff for sea cucumber containing asthma          improving substance and manufacturing method <130> PD18-5660 <160> 4 <170> KoPatentIn 3.0 <210> 1 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> GAPDH F <400> 1 tacccccaat gtgtccgtc 19 <210> 2 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> GAPDH R <400> 2 aagagtggga gttgctgttg aag 23 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> IL-10 F <400> 3 gctatgctgc ctggtcttac tg 22 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> IL-10 R <400> 4 tccagctggt cctttgtttg 20

Claims (2)

갈파래, 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상의 조류의 분말 10 내지 20 중량부, 지충이(Sargassum thunbergii) 10 내지 20 중량부, 대두박(soybean meal) 5 내지 10 중량부, 조개분말(shell fish powder) 5 내지 10 중량부, 패각분말(Shell powder) 1 내지 3 중량부, 인산칼슘(calcium phosphate) 1 내지 3 중량부, 효모 추출물(yeast extract) 3 내지 8 중량부, 콩 레시틴(soyabean lecithin) 2 내지 6 중량부, 미네랄(mineral) 0.2 내지 0.7 중량부, 비타민(vitamin) 0.2 내지 0.7 중량부, 및 바다 진흙(sea mud) 20 내지 60 중량부를 포함하는, 천식 개선 효과가 강화된 해삼의 생산을 위한 해삼 양식 사료조성물. 10-20 parts by weight of powder of one or more algae selected from the group consisting of brown seaweed, kelp and Nannochloropsi s sp., 10-20 parts by weight of Sargassum thunbergii , soybean meal 5 to 10 parts by weight, shell fish powder 5 to 10 parts by weight, shell powder 1 to 3 parts by weight, calcium phosphate 1 to 3 parts by weight, yeast extract 3 to 8 parts by weight, 2 to 6 parts by weight of soyabean lecithin, 0.2 to 0.7 parts by weight of mineral, 0.2 to 0.7 parts by weight of vitamin, and 20 to 60 parts by weight of sea mud Contains, sea cucumber culture feed composition for the production of sea cucumber enhanced asthma effect. 갈파래, 다시마 및 나노클롭시스 속(Nannochloropsis sp.)으로 구성되는 군으로부터 선택되는 하나 또는 둘 이상 조류의 분말 10 내지 20 중량부, 지충이(Sargassum thunbergii) 10 내지 20 중량부, 대두박(soybean meal) 5 내지 10 중량부, 조개분말(shell fish powder) 5 내지 10 중량부, 패각분말(Shell powder) 1 내지 3 중량부, 인산칼슘(calcium phosphate) 1 내지 3 중량부, 효모 추출물(yeast extract) 3 내지 8 중량부, 콩 레시틴(soyabean lecithin) 2 내지 6 중량부, 미네랄(mineral) 0.2 내지 0.7 중량부, 비타민(vitamin) 0.2 내지 0.7 중량부, 및 바다 진흙(sea mud) 20 내지 60 중량부를 혼합하여 혼합물을 제조하는 단계; 및
상기 혼합물을 건조시켜 고형화하는 단계를 포함하는 천식 개선 효과가 강화된 해삼 생산을 위한 해삼 양식용 사료조성물의 제조방법.10 to 20 parts by weight of powder of one or more algae selected from the group consisting of brown seaweed, kelp and Nannochloropsi s sp., 10 to 20 parts by weight of Sargassum thunbergii , soybean meal 5 to 10 parts by weight, shell fish powder 5 to 10 parts by weight, shell powder 1 to 3 parts by weight, calcium phosphate 1 to 3 parts by weight, yeast extract 3 to 8 parts by weight, 2 to 6 parts by weight of soyabean lecithin, 0.2 to 0.7 parts by weight of mineral, 0.2 to 0.7 parts by weight of vitamin, and 20 to 60 parts by weight of sea mud Mixing to prepare a mixture; And
Method for producing a sea cucumber culture feed composition for producing sea cucumber enhanced asthma effect comprising the step of drying the mixture to solidify.
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