KR102598346B1 - Feed additive for reducing methane gas in rumen, and feed composition for ruminant containing same - Google Patents
Feed additive for reducing methane gas in rumen, and feed composition for ruminant containing same Download PDFInfo
- Publication number
- KR102598346B1 KR102598346B1 KR1020230011196A KR20230011196A KR102598346B1 KR 102598346 B1 KR102598346 B1 KR 102598346B1 KR 1020230011196 A KR1020230011196 A KR 1020230011196A KR 20230011196 A KR20230011196 A KR 20230011196A KR 102598346 B1 KR102598346 B1 KR 102598346B1
- Authority
- KR
- South Korea
- Prior art keywords
- methane
- feed
- rumen
- attapulgite
- methane gas
- Prior art date
Links
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Classifications
-
- 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
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/28—Silicates, e.g. perlites, zeolites or bentonites
-
- 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
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/202—Algae extracts
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
- A23V2250/212—Garlic
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
- A23V2250/214—Tea
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/22—Methane [CH4], e.g. from rice paddies
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Physiology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Inorganic Chemistry (AREA)
- Birds (AREA)
- Fodder In General (AREA)
- Feed For Specific Animals (AREA)
Abstract
본 발명은 반추위 내 메탄가스 저감을 위한 사료 첨가제 및 이를 함유한 반추동물용 사료 조성물에 관한 것으로서, 보다 구체적으로, 마늘추출물과 아타풀자이트를 포함하는, 반추위 내 메탄가스 저감을 위한 사료 첨가제, 및 이러한 반추위 내 메탄가스 저감을 위한 사료 첨가제를 포함하는 반추 동물용 사료 조성물에 관한 것이다.
본 발명의 반추위 내 메탄가스 저감을 위한 사료 첨가제 및 이를 함유한 반추동물용 사료 조성물은 반추위 내 메탄 가스 생성을 억제하여 온실가스 효과 감소 에 의한 대기환경 문제를 해결하는 효과를 나타낸다. 또한, 발명의 사료 첨가제 및 이를 함유한 반추동물용 사료 조성물은 반추위 내 메탄가스를 저감하는 바, 사료 에너지의 사료 효율을 높여, 사료 에너지가 대부분 반추동물의 성장에 이용될 수 있다.The present invention relates to a feed additive for reducing methane gas in the rumen and a feed composition for ruminants containing the same. More specifically, a feed additive for reducing methane gas in the rumen, comprising garlic extract and attapulgite, and It relates to a feed composition for ruminants containing a feed additive for reducing methane gas in the rumen.
The feed additive for reducing methane gas in the rumen of the present invention and the feed composition for ruminants containing the same have the effect of suppressing the production of methane gas in the rumen and solving atmospheric environmental problems by reducing greenhouse gas effects . In addition, the feed additive of the invention and the feed composition for ruminants containing the same reduce methane gas in the rumen, thereby increasing feed efficiency of feed energy, so that most of the feed energy can be used for the growth of ruminants.
Description
본 발명은 반추동물을 위한 사료 첨가제 및 이를 함유한 사료 조성물에 관한 것으로서, 구체적으로, 반추위 내 메탄가스 저감을 위한 사료 첨가제 및 이를 함유한 반추동물용 사료 조성물에 관한 것이다.The present invention relates to a feed additive for ruminants and a feed composition containing the same. Specifically, it relates to a feed additive for reducing methane gas in the rumen and a feed composition for ruminants containing the same.
국제적인 산업화 영향으로 온실가수 배출량은 지속적으로 증가하고 있어, 심각한 지구 온난화를 야기하고 있다. 세계 온도는 최근 100년간 약 1도 가량 상승하였으며 상승폭이 점점 더 가속화되어서 현재 추세를 이어갈 경우 2040년 이전에 1.5도 상승할 것으로 예측된다. 특히 메탄은 이산화탄소와 비교하여 25 내지 30배의 온실효과를 발생하는 것으로 알려져 있다[IPCC, 2001]. Due to the influence of international industrialization, greenhouse gas emissions are continuously increasing, causing serious global warming. The world's temperature has risen by about 1 degree over the past 100 years, and the rate of increase is expected to accelerate by 1.5 degrees before 2040 if the current trend continues. In particular, methane is known to produce a greenhouse effect 25 to 30 times greater than that of carbon dioxide [IPCC, 2001].
이에 1992년에 선진국 위주로 지구 온난화에 대응하기 위한 국제 협의체가 형성되어 온실가스 배출을 저감하기 위해 구체적인 실행 방안과 노력이 지속되고 있다. 121개 국가가 기후목표 상향 동맹에 가입하고, 2021년 4월에 열린 기후정상회의를 계기로 미국, EU, 영국, 일본 등이 2050 탄소중립 목표에 맞추어 상향된 국가 온실가스 감축목표를 발표하였다.Accordingly, in 1992, an international consultative body was formed mainly in developed countries to respond to global warming, and specific action plans and efforts are continuing to reduce greenhouse gas emissions. 121 countries joined the Alliance to Raise Climate Goals, and at the Climate Summit held in April 2021, the US, EU, UK, Japan, etc. announced raised national greenhouse gas reduction goals in line with the 2050 carbon neutrality goal.
우리나라는 또한 2050 탄소중립 시나리오를 수립하였으며 탄소배출량과 탄소흡수량을 맞추어서 순(Net) 배출량을 0(zero)으로 만드는 것을 목표로 하고 있다. 이를 위해 각 산업 분야에서 할당된 목표를 달성하고자 하며, 구체적으로 농축수산 부문의 탄소 발생량은 2018년 24.7 백만톤에서 15.4백만톤으로 37.7% 감안하는 것을 목표로 하며, 그 중 가축의 장내발효에 의하여 생산되는 메탄은 2018년 4.5백만 톤에서 2050년 2.8백만톤으로, 반추위내의 약 1.7백만톤 메탄 감축을 목표로 한다.Korea has also established a 2050 carbon neutral scenario and aims to reduce net emissions to zero by matching carbon emissions and carbon absorption. To this end, we aim to achieve the assigned goals in each industry sector, and specifically, we aim to account for 37.7% of the carbon emissions in the agricultural, livestock and fisheries sector from 24.7 million tons to 15.4 million tons in 2018, of which the amount is due to enteric fermentation of livestock. The methane produced will increase from 4.5 million tons in 2018 to 2.8 million tons in 2050, with the goal of reducing about 1.7 million tons of methane in the rumen.
반추위 내에서 메탄을 발생하는 메탄생성균은 분류학적으로 일반세균(b acteria)과는 근본적으로 다른 고세균(archaea)에 속하는 미생물이다. 세균은 펩티도글리칸 층을, 고균은 슈도펩티도글리칸층을 이루는 것과 같이 물질대사에 관여하는 효소체계나, 세포벽의 구성물질처럼, 일반세균과는 매우 다르다. 기존의 연구에 의하면 메탄생성 고세균은 세 개의 분류군으로 구성되어 있는데, 젖소의 경우 메타노브레비박테르(Methanobrevibacter), 메타노메실리코커스, 메타노스피에라 속 그룹으로 분류되고 있다(한국 공학 생명연구원 et al., 2017)Methanogenic bacteria, which produce methane in the rumen, are taxonomically microorganisms belonging to archaea, which are fundamentally different from general bacteria (b acteria). Bacteria have a peptidoglycan layer, and archaea have a pseudopeptidoglycan layer, and the enzyme systems involved in metabolism and the components of the cell wall are very different from ordinary bacteria. According to previous studies, methanogenic archaea are composed of three taxa, and in the case of dairy cattle, they are classified into the genus Methanobrevibacter , Methanomesilicoccus, and Methanospiera (Korea Institute of Engineering and Life Science et al. ., 2017)
이처럼 가축에서 발생된 메탄은 지구온난화에 영향을 줄 뿐만 아니라, 장내 발효에 의한 메탄 가스의 생성과 배출은 반추 동물이 사료로부터 얻는 총 에너지의 2 내지 12% 정도의 손실이기 때문에 메탄 가스 생성으로 인해 사료 이용 효율의 감소와 생산성의 감소가 야기될 수 있다(Johnson et al., 1991; Ha et al., 2005). 즉 반추 동물의 메탄 배출량을 감소시킨다면 지구온난화를 늦추는 환경적 측면과 사료 에너지의 사료 효율을 높이는 영양학적 측면을 향상 시킬 수 있는 일석이조의 효과가 나타낼 수 있다(Lee and Ha, 2009).In this way, methane generated from livestock not only affects global warming, but the production and emission of methane gas through intestinal fermentation is a loss of about 2 to 12% of the total energy that ruminants obtain from feed, so the production of methane gas causes This may result in reduced feed use efficiency and reduced productivity (Johnson et al., 1991; Ha et al., 2005). In other words, reducing methane emissions from ruminants can kill two birds with one stone by improving the environmental aspect of slowing global warming and the nutritional aspect of increasing feed efficiency of feed energy (Lee and Ha, 2009).
장내발효를 통한 메탄발생 저감기술 개발은 전세계적으로 “첨가제 개발”과 “사료 개발”의 두 방향에 초점이 맞춰져 있지만 아직 지속적인 현장 적용기술이 개발되어 있지 않은 상태이며, 크게 아래 4가지 방법 등으로 요약할 수 있다.The development of technology to reduce methane production through enteric fermentation is focused around the world on two directions: “additive development” and “feed development,” but continuous field application technology has not yet been developed. It can be broadly divided into the four methods below. It can be summarized.
① 메탄가스 생성의 원인이 되는 메탄생성 미생물을 직접적으로 억제하는 방법 ① Method of directly suppressing methane-producing microorganisms that cause methane gas production
② 메탄가스의 원인이 되는 수소의 발생을 막아 간접적으로 메탄생성균을 억제하는 방법 ② A method of indirectly suppressing methane-producing bacteria by preventing the generation of hydrogen, which causes methane gas.
③ 메탄 발효 외에 Propionate발효 등 수소를 소모하는 다른 방법을 촉진하여 간접적으로 메탄생성균을 억제하는 방법 ③ A method of indirectly suppressing methane-producing bacteria by promoting other methods of consuming hydrogen, such as propionate fermentation, in addition to methane fermentation.
④ 반추위 미생물의 군집(다양성)을 바꾸어 메탄균 활성을 줄이는 방법이 있다. ④ There is a way to reduce methane bacteria activity by changing the community (diversity) of rumen microorganisms.
현재까지 개발된 메탄저감 사료첨가제는 해초류, 질산염, 3-nop, 항생제 등 화학제품과 단일물질의 기반으로 단순 급여를 통하여 메탄 발생을 억제하였다. 그러나, 화학제품이나 항생제의 직접 급여 등은 반추동물 개체에 독성을 유발하고, 2차 생산 물질에 잔류되거나 내성문제가 발생되어, 기본적 생리를 방해한다는 문제점이 있다. 단일 물질의 급여가 메탄 저감에 효과는 좋을 수 있으나 반추 위의 발효 성상을 바꾸고 소의 생리 균형을 무너뜨리는 문제가 발생한다고 판단하여, 기호성 문제와 반추동물의 생리 문제를 상쇄시킬 수 있는 복합 사료 첨가제의 개발을 하고자 하였으며 해당 개발 기술이 기호성 향상과 발효 성상 밸런스 유지를 도울 것으로 예상한다. 따라서, 온실가스의 한 종류인 메탄이 반추 동물의 반추위 내에서 생성되는 것을 억제하면서 인체나 동물에 해를 주지 않는 물질이 요구되고 있다.Methane-reducing feed additives developed to date are based on chemical products and single substances such as seaweed, nitrate, 3-nop, and antibiotics, and suppress methane generation through simple feeding. However, direct feeding of chemical products or antibiotics has the problem of causing toxicity to ruminant animals, remaining in secondary production materials or causing resistance problems, thereby interfering with basic physiology. Feeding a single substance may be effective in reducing methane, but it is believed that it causes problems by changing the nature of fermentation in the ruminant stomach and disrupting the physiological balance of cattle, so a complex feed additive that can offset the palatability problem and physiological problems of ruminants is recommended. We wanted to develop it, and it is expected that the development technology will help improve palatability and maintain the balance of fermentation characteristics. Therefore, there is a need for a material that suppresses the production of methane, a type of greenhouse gas, in the rumen of ruminants and does not harm humans or animals.
선행기술문헌Prior art literature
특허문헌patent literature
(특허문헌 001) 대한민국 특허출원 제2021-0189023호(Patent Document 001) Republic of Korea Patent Application No. 2021-0189023
(특허문헌 002) 대한민국 특허출원 제2019-0177357호(Patent Document 002) Republic of Korea Patent Application No. 2019-0177357
(특허문헌 003) 대한민국 특허출원 제2018-0061694호(Patent Document 003) Republic of Korea Patent Application No. 2018-0061694
(특허문헌 004) 대한민국 특허출원 제2017-0101990호(Patent Document 004) Republic of Korea Patent Application No. 2017-0101990
본 발명의 목적은 천연 물질을 포함하는 반추동물의 반추위 내 메탄가스 저감을 위한 사료 첨가제를 제공하는 데 있다.The purpose of the present invention is to provide a feed additive for reducing methane gas in the rumen of ruminants containing natural substances.
또한, 본 발명의 다른 목적은 반추동물의 반추위 내 메탄가스 저감을 위한 상기 사료 첨가제를 함유하는 반추동물용 사료 조성물을 제공하는 데 있다.In addition, another object of the present invention is to provide a feed composition for ruminants containing the feed additive for reducing methane gas in the rumen of ruminants.
상기한 목적을 달성하기 위해, 본 발명은 천연물질로서 마늘추출물과 아타풀자이트를 포함하는, 반추위 내 메탄가스 저감을 위한 사료 첨가제를 제공한다.In order to achieve the above object, the present invention provides a feed additive for reducing methane gas in the rumen, which contains garlic extract and attapulgite as natural substances.
바람직하게는, 상기 반추위 내 메탄가스 저감을 위한 사료 첨가제는 미역박, 녹차박, 린씨드 및 감피 중 하나 이상을 추가로 포함할 수 있다.Preferably, the feed additive for reducing methane gas in the rumen may further include one or more of seaweed meal, green tea meal, linseed, and persimmon peel.
바람직하게는, 상기 사료 첨가제에서 상기 마늘추출물 및 아타풀자이트의 중량비는 0.5 내지 5.5:0.5 내지 5.5일 수 있다.Preferably, the weight ratio of the garlic extract and attapulgite in the feed additive may be 0.5 to 5.5:0.5 to 5.5.
바람직하게는, 상기 사료 첨가제에서 상기 마늘추출물, 아타풀자이트, 및 미역박, 녹차박, 린씨드 및 감피 중 하나 이상의 중량비는 0.5 내지 5.5:0.5 내지 5.5:0.5 내지 5.5일 수 있다.Preferably, in the feed additive, the weight ratio of the garlic extract, attapulgite, and one or more of seaweed meal, green tea meal, linseed, and persimmon may be 0.5 to 5.5:0.5 to 5.5:0.5 to 5.5.
또한, 상기한 다른 목적을 달성하기 위해, 본 발명은 마늘추출물과 아타풀자이트를 포함하는 반추위 내 메탄가스 저감을 위한 사료 첨가제를 포함하는, 반추 동물용 사료 조성물을 제공한다.In addition, in order to achieve the above-described other objects, the present invention provides a feed composition for ruminants, including a feed additive for reducing methane gas in the rumen, including garlic extract and attapulgite.
바람직하게는, 상기 반추위 내 메탄가스 저감을 위한 사료 첨가제는 미역박, 녹차박, 린씨드 및 감피 중 하나 이상을 추가로 포함할 수 있다.Preferably, the feed additive for reducing methane gas in the rumen may further include one or more of seaweed meal, green tea meal, linseed, and persimmon peel.
바람직하게는, 사료 조성물 100 중량%를 기준으로 상기 마늘추출물 및 아타풀자이트의 양은 각각 0.5 내지 5.5 중량%일 수 있다.Preferably, based on 100% by weight of the feed composition, the amounts of the garlic extract and attapulgite may each be 0.5 to 5.5% by weight.
바람직하게는, 상기 사료 조성물 100 중량%를 기준으로 상기 마늘추출물의 양은 0.5 내지 2.5 중량%일 수 있으며, 상기 아타풀자이트의 양은 2.5 내지 5.5 중량%일 수 있다.Preferably, based on 100% by weight of the feed composition, the amount of the garlic extract may be 0.5 to 2.5% by weight, and the amount of attapulgite may be 2.5 to 5.5% by weight.
바람직하게는, 상기 사료 조성물 100 중량%를 기준으로 상기 마늘추출물의 양은 2.5 중량%일 수 있으며, 상기 아타풀자이트의 양은 5.5 중량%일 수 있다.Preferably, based on 100% by weight of the feed composition, the amount of the garlic extract may be 2.5% by weight, and the amount of the attapulgite may be 5.5% by weight.
바람직하게는, 상기 사료 조성물 100 중량%를 기준으로 상기 미역박, 녹차박, 린씨드 및 감피 중 하나 이상의 양이 0.5 내지 5.5 중량%일 수 있다.Preferably, based on 100% by weight of the feed composition, the amount of one or more of the seaweed meal, green tea meal, linseed, and persimmon may be 0.5 to 5.5% by weight.
본 발명의 반추위 내 메탄가스 저감을 위한 사료 첨가제 및 이를 함유한 반추동물용 사료 조성물은 반추위 내 메탄 가스 생성을 억제하여 지구 온난화를 늦추는 효과를 나타낸다.The feed additive for reducing methane gas in the rumen of the present invention and the feed composition for ruminants containing the same have the effect of slowing global warming by suppressing the production of methane gas in the rumen.
특히, 반추위 내에서 메탄가스의 발생량이 많다는 것은 급여된 사료의 에너지 일부분이 메탄가스로 대기 중에 방출된다는 것을 의미하므로, 메탄가스 발생량이 많으면 반추동물의 생산성도 저하된다. 그러나, 본 발명의 사료 첨가제 및 이를 함유한 반추동물용 사료 조성물은 반추위 내 메탄가스를 저감하는 바, 사료 에너지의 사료 효율을 높여, 사료 에너지가 대부분 반추동물의 성장에 이용될 수 있다.In particular, the large amount of methane gas generated in the rumen means that part of the energy of the fed feed is released into the atmosphere as methane gas, so if the amount of methane gas generated is large, the productivity of ruminant animals also decreases. However, the feed additive of the present invention and the feed composition for ruminants containing the same reduce methane gas in the rumen, thereby increasing feed efficiency of feed energy, so that most of the feed energy can be used for the growth of ruminants.
도 1은 메탄 발생량(mL/g DM)에 대한 알리신의 효능을 도시한 그래프이다.
도 2는 메탄 발생량(mL/g DM)에 대한 아타풀자이트의 효능을 도시한 그래프이다.
도 3a는 본 발명에서 사용되는 메탄 측정기기인 레이저 메탄 검출기(kyewon kang et al. 2020)의 개략도이다.
도 3b는 메탄 측정기기를 이용하여 0.5초 간격으로 6분간 측정한 결과를 나타낸 그래프이다.
도 4는 4주 동안 대조군과 실험군의 동물에서 배출하는 메탄 발생량을 도시한 그래프이다.Figure 1 is a graph showing the efficacy of allicin on methane production (mL/g DM).
Figure 2 is a graph showing the efficacy of attapulgite on methane production (mL/g DM).
Figure 3a is a schematic diagram of a laser methane detector (kyewon kang et al. 2020), a methane measurement device used in the present invention.
Figure 3b is a graph showing the results of measurements for 6 minutes at 0.5 second intervals using a methane measuring device.
Figure 4 is a graph showing the amount of methane emissions from animals in the control and experimental groups over 4 weeks.
일 양태에서, 본 발명은 천연 물질인 알리신과 아타풀자이트를 포함하는, 반추위 내 메탄가스 저감을 위한 사료 첨가제를 제공한다.In one aspect, the present invention provides a feed additive for reducing methane gas in the rumen, including the natural substances allicin and attapulgite.
통상적으로, 반추 동물은 우제목/경우제목에 속하는 동물로 반추위를 가지고 있으며, 한번 삼킨 먹이를 다시 게워 내어 씹는 특성을 가지고 있다. 되새김 동물이라고도 하고, 반추류와 낙타아목의 동물을 포함하고, 소, 양, 염소, 버팔로, 사슴, 카멜리드(camelid), 지라피드(giraffid), 영양, 들소 및 야크로 구성되는 군으로부터 선택될 수 있다. Typically, ruminants are animals belonging to the order Artiodactyla/Arminozoa, have a rumen, and have the characteristic of regurgitating and chewing food once swallowed. Also called ruminating animals, they include animals of the ruminant and camelid orders and may be selected from the group consisting of cattle, sheep, goats, buffalo, deer, camelids, giraffids, antelopes, bison and yaks. You can.
본 발명의 사료 첨가제의 하나의 천연물질 성분인 알리신(allicin)은 마늘이나 양파에 함유된 성분으로, 강한 냄새를 가지는 유황 화합물, 황화아릴의 일종이다. 일반적으로, 알리신이 함유된 식품에는 마늘, 양파, 대파, 부추 등이 있다. 알리신은 조효소 활성을 저해하거나, 항미생물 작용이 있다. 항균에 대한 메커니즘은 두 가지가 있는데, 마늘이 활성산소를 생산하게 하여 세포로 하여금 산화스트레스를 받아 사멸하게 하거나 또는 산화환원의 완충작용을 통해 세포를 보호하는 세포내 -SH기 물질과 반응하여 세포막 구조를 손상시키고, 아포토시스를 유도할 수 있다. 이는, 세포막에 빠르게 침투하여 미생물의 티올 단백질과 결합하여 신진대사를 억제하며 알리신의 S-알릴 부분(S-CH 2 -CHCH 2 )은 대사에 필수적인 효소 단백질의 -SH기와 반응하여 불활성시키게 되는 것이다. 특정한 세균을 제외하고는 항세균작용을 기대할 수 없으며 메탄을 생성하는 그람양성세균(Methanobacterium)은 그람음성세균보다, 직접적인 영향으로 항균작용을 나타낸다[Ziletao Tao.,2020; Ziletao Tao.,2020]. 이처럼 복합사료첨가제의 항미생물 작용으로 메탄을 생성하는 메탄생성균을 직접적으로 제어할 수 있다. Allicin, a natural component of the feed additive of the present invention, is a component contained in garlic or onions and is a type of aryl sulfide, a sulfur compound with a strong odor. Generally, foods containing allicin include garlic, onions, green onions, and chives. Allicin inhibits coenzyme activity or has antimicrobial effects. There are two mechanisms for antibacterial activity: Garlic produces active oxygen, which causes cells to die from oxidative stress, or it reacts with intracellular -SH substances that protect cells through redox buffering, destroying the cell membrane. It can damage the structure and induce apoptosis. It quickly penetrates the cell membrane and binds to the thiol protein of microorganisms to inhibit metabolism, and the S-allyl part (S-CH 2 -CHCH 2) of allicin reacts with the -SH group of the enzyme protein essential for metabolism and becomes inactive. . Except for certain bacteria, antibacterial activity cannot be expected, and methane-producing gram-positive bacteria (Methanobacterium) exhibit antibacterial activity through a more direct effect than gram-negative bacteria [Ziletao Tao., 2020; Ziletao Tao.,2020]. In this way, the antimicrobial action of the complex feed additive can directly control methanogenic bacteria that produce methane.
또한, 본 발명의 사료 첨가제 중 다른 한 가지 천연물질 성분인 아타풀자이트는 지구상 광물 중에 기공성이 가장 큰 광물로 흡착 능력을 가지고 있고, 강 하류의 퇴적황토를 분류하여 제조한다. 동물 사료에 보충제로 사용되는 점토 광물로 다른 점토에 비해 독특한 구조로 2:1 필로실리케이트(phyllosilicate)의 리본으로 구성되어 있으며, 각각의 리본은 Si-O-Si 결합 세트를 따라 SiO4 사면체의 반전에 의해 다른 하나와 연결된다. 이러한 구조는 섬유형태를 초래하며 높은 점도와 독소를 바인더 하는 흡착능력을 가지고 있다. 통상적인 효과로는 독소사람과 동물 모두 자연적으로 설사를 예방하는 특성을 가지고 있고, 염증을 줄이며 항산화 능력을 향상시킨다. 아타풀자이트는 메탄 가스를 직접 흡착하여 감소시키고 동시에 Ca2+, Mg2+, K+, Fe3+ 금속 이온을 방출하여 가수분해와 메탄 생성 단계의 효소 활성 감소 효과를 가져올 수 있다.In addition, attapulgite, another natural component of the feed additive of the present invention, is a mineral with the highest porosity among minerals on Earth and has an adsorption capacity, and is manufactured by fractionating sedimentary loess from the lower reaches of rivers. A clay mineral used as a supplement in animal feed. It has a unique structure compared to other clays and is composed of 2:1 ribbons of phyllosilicate, each ribbon being an inversion of a SiO 4 tetrahedron along a set of Si-O-Si bonds. connected to the other by This structure results in a fibrous form and has high viscosity and adsorption capacity to bind toxins. Common effects include: It has natural diarrhea-preventing properties in both humans and animals, reduces inflammation, and improves antioxidant capacity. Attapulgite directly adsorbs and reduces methane gas and simultaneously releases Ca 2+ , Mg 2+ , K + , and Fe 3+ metal ions, which can reduce enzyme activity in the hydrolysis and methane production stages.
본 발명의 사료 첨가제에서 알리신 및 아타풀자이트의 중량비는 0.5 내지 5.5:0.5 내지 5.5일 수 있고, 바람직하게는, 0.5 내지 2.5:2.5 내지 5.5일 수 있고, 더욱 바람직하게는, 2.5:5.5일 수 있다. 상기 범위를 벗어나는 경우, 반추위 내 메탄가스 저감 효과가 떨어지는 단점을 나타내어, 본 발명의 최상의 작용 효과를 얻을 수 없다.The weight ratio of allicin and attapulgite in the feed additive of the present invention may be 0.5 to 5.5:0.5 to 5.5, preferably 0.5 to 2.5:2.5 to 5.5, and more preferably 2.5:5.5. there is. If it is outside the above range, the effect of reducing methane gas in the rumen is reduced, and the best effect of the present invention cannot be obtained.
본 발명의 사료 첨가제에는 알리신 및 아타풀자이트 이외에, 미역박, 녹차박, 린씨드 및 감피 중 하나 이상을 추가로 포함할 수 있다.In addition to allicin and attapulgite, the feed additive of the present invention may further include one or more of seaweed meal, green tea meal, linseed, and gampi.
미역박은 일반적인 미역원료 세척을 통해 잡질, 불순물 제거 후 수침과 압착을 통해 염분과 수분을 제거하고, 절단, 선별을 통해 불가식 부위를 확보하여 인공건조 및 분쇄를 통해 확보할 수 있으며, 이러한 미역박에는 브로모포름 등의 할로겐 화합물 성분이 포함되어 있다.Seaweed meal can be obtained by washing general seaweed raw materials to remove contaminants and impurities, removing salt and moisture through soaking and pressing, securing inedible parts through cutting and sorting, and artificially drying and crushing them. Contains halogenated compounds such as bromoform.
또한, 녹차박, 린씨드 및 감피는 통상적으로 사용되는 것으로서 자연에서 얻은 원료를 적절하게 가공 및 세척하여 얻을 수 있거나, 이를 상업적으로 입수 가능하다.In addition, green tea peel, linseed, and persimmon are commonly used and can be obtained by appropriately processing and washing raw materials obtained from nature, or they can be obtained commercially.
이러한 추가 성분을 함유하는 경우, 알리신, 아타풀자이트, 및 미역박, 녹차박, 린씨드 및 감피 중 하나 이상의 중량비는 0.5 내지 5.5:0.5 내지 5.5:0.5 내지 5.5인 것이 바람직하며, 상기 범위를 벗어나는 경우, 반추위 내 메탄가스 저감 효과가 떨어지는 단점을 나타내어, 본 발명의 최상의 작용 효과를 얻을 수 없다.When containing such additional ingredients, the weight ratio of allicin, attapulgite, and one or more of seaweed meal, green tea meal, linseed, and persimmon is preferably 0.5 to 5.5:0.5 to 5.5:0.5 to 5.5, and the weight ratio outside the above range is preferably In this case, the effect of reducing methane gas in the rumen is low, and the best effect of the present invention cannot be obtained.
이러한 반추위 내 메탄가스 저감을 위한 사료 첨가제는 하기 실시예 및 실험예를 통하여 알 수 있는 바와 같이, 천연 물질들을 최적의 비율로 조합함으로써 단일 물질의 문제를 서로 보완하고 우수한 반추위 내의 메탄 저감 효과를 나타낼 수 있다.As can be seen through the examples and experimental examples below, the feed additive for reducing methane gas in the rumen complements the problems of single substances by combining natural substances in optimal ratios and exhibits an excellent methane reduction effect in the rumen. You can.
또한, 다른 양태에서, 본 발명은 알리신과 아타풀자이트를 포함하는 반추위 내 메탄가스 저감을 위한 사료 첨가제를 포함하는, 반추 동물용 사료 조성물을 제공한다.In addition, in another aspect, the present invention provides a feed composition for ruminants, including a feed additive for reducing methane gas in the rumen, including allicin and attapulgite.
본 발명에서 반추 동물용 사료 조성물은 통상적으로 사용되는 모든 반추 동물용 사료 조성물을 사용할 수 있으며, 일 예로 조사료, 농후사료, 보조사료, 배합사료 등을 적절한 비율로 혼합하여 얻어진 사료 조성물일 수 있으며, 본 발명에서는 이의 구체적인 성분 및 비율로 제한되지 않는다.In the present invention, the feed composition for ruminants can be any commonly used feed composition for ruminants. For example, it may be a feed composition obtained by mixing roughage, concentrated feed, supplementary feed, compound feed, etc. in an appropriate ratio, The present invention is not limited to its specific components and ratios.
본 발명의 반추 동물용 사료 조성물의 성분인 알리신과 아타풀자이트의 양은 본 발명의 사료 조성물 100 중량%를 기준으로 각각 0.5 내지 5.5 중량%일 수 있고, 바람직하게는, 사료 조성물 100 중량%를 기준으로 알리신의 양은 0.5 내지 2.5 중량%일 수 있으며, 아타풀자이트의 양은 2.5 내지 5.5 중량%일 수 있고, 더욱 바람직하게는, 사료 조성물 100 중량%를 기준으로 알리신의 양은 2.5 중량%일 수 있으며, 아타풀자이트의 양은 5.5 중량%일 수 있다.The amounts of allicin and attapulgite, which are components of the feed composition for ruminants of the present invention, may each be 0.5 to 5.5% by weight based on 100% by weight of the feed composition of the present invention, preferably, based on 100% by weight of the feed composition. The amount of allicin may be 0.5 to 2.5% by weight, the amount of attapulgite may be 2.5 to 5.5% by weight, and more preferably, the amount of allicin may be 2.5% by weight based on 100% by weight of the feed composition. The amount of attapulgite may be 5.5% by weight.
상기 범위의 알리신 및 아타풀자이트를 포함하는 경우, 최상의 반추위 내 메탄가스 저감 효과를 나타낼 수 있으며, 상기 범위를 벗어나는 경우에는 이러한 효과를 크게 떨어지는 단점을 나타낸다.When allicin and attapulgite are included in the above range, it can exhibit the best effect of reducing methane gas in the rumen, and when it is outside the above range, this effect is significantly reduced.
또한, 반추위 내 메탄가스 저감을 위한 사료 첨가제는 알리신과 아타풀자이트 이외에, 미역박, 녹차박, 린씨드 및 감피 중 하나 이상을 추가로 포함할 수 있고, 바람직하게는, 상기 사료 조성물 100 중량%를 기준으로 0.5 내지 5.5 중량% 양의 미역박, 녹차박, 린씨드 및 감피 중 하나 이상을 추가로 포함할 수 있다.In addition, the feed additive for reducing methane gas in the rumen may further include one or more of seaweed meal, green tea meal, linseed, and persimmon peel in addition to allicin and attapulgite, and preferably, 100% by weight of the feed composition. It may additionally include one or more of seaweed meal, green tea meal, linseed, and gampi in an amount of 0.5 to 5.5% by weight.
상기 범위에 포함되는 경우, 최상의 반추위 내 메탄가스 저감 효과를 나타내기 위한 시너지 효과를 나타낼 수 있고 반추동물의 영양학적 측면에서 우수한 효과를 나타낼 수 있으며, 상기 범위를 벗어나는 경우에는 이러한 효과를 크게 떨어지는 단점을 나타낸다.If it is within the above range, it can exhibit a synergistic effect to show the best methane gas reduction effect in the rumen and can have an excellent effect in terms of nutrition for ruminants, but if it is outside the above range, this effect is greatly reduced. represents.
본 발명의 반추동물용 사료 조성물은 하기 실험예 4에 기술되는 바와 같이, 반추위내 메탄 발생량 중 호흡과 트름을 분리하여 계산하였을 때 첨가제 처리군과 대조군의 차이는 통계적으로 유의미함을 나타내었다(P<0.05) 이는 아타풀자이트와 알리신의 복합 첨가제는 항메타노젠과 메탄 발생 효소를 저해한다는것으로 해석할 수 있다. 보다 구체적으로 도 4 및 표 19에 도시된 바와 같이, 소의 트름의 메탄 발생량은(ppm/m) 1주차 40.79%> 2주차 32.80%> 3주차 45.83%, 호흡의 메탄 발생량은 1주차 13.82%>2주차 39.62%> 3주차 57.72% 로 감소한 것으로 검증되었다. As described in Experimental Example 4 below, the feed composition for ruminants of the present invention showed that the difference between the additive treatment group and the control group was statistically significant when respiration and belching were calculated separately among the amount of methane generated in the rumen (P <0.05) This can be interpreted that the complex additive of attapulgite and allicin inhibits anti-methanogen and methanogenic enzymes. More specifically, as shown in Figure 4 and Table 19, the methane production amount from cow's burp (ppm/m) is 40.79% in the first week > 32.80% in the second week > 45.83% in the third week, and the methane production amount in respiration is > 13.82% in the first week. It was verified that it decreased from 39.62% in the 2nd week to 57.72% in the 3rd week.
메탄 발생량을 시간에 흐름에 따라, 호흡은 1주차에는 메탄감소의 경향성이 있으며 2주차부터 유의미적으로 감소되는것을 확인할 수 있었고(p<0.05), 트름은 1주차부터 감소하여 급여하는 기간이 늘어날수록 메탄 저감 효과가 더욱 큰 것을 확인할 수 있었다(p<0.001). 즉 대조군에 비하여 모든 처리군은 메탄 생성량이 감소하는 결과를 우수한 결과를 보였다. Depending on the amount of methane production over time, respiration tended to decrease in methane in the first week and significantly decreased from the second week (p<0.05), and burping decreased from the first week, increasing the feeding period. It was confirmed that the methane reduction effect was greater (p<0.001). In other words, compared to the control group, all treatment groups showed excellent results in reducing methane production.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시하나, 하기 실시예는 본 발명을 예시하는 것일 뿐 본 발명의 범주 및 기술사상 범위 내에서 다양한 변경 및 수정이 가능함은 당업자에게 있어서 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속하는 것도 당연한 것이다.Hereinafter, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are merely illustrative of the present invention, and it is clear to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention. It is natural that such variations and modifications fall within the scope of the attached patent claims.
<실시예> 메탄가스 저감을 위한 사료 조성물의 제조<Example> Preparation of feed composition for reducing methane gas
사료 조성물에 상기 표 1에 기재된 성분들을 총 5% DM 기준(Isaac A. Aboagye, 2019)으로 첨가 및 혼합하여, 본 발명의 사료 조성물을 제조하였다.The feed composition of the present invention was prepared by adding and mixing the ingredients listed in Table 1 to the feed composition at a total of 5% DM (Isaac A. Aboagye, 2019).
비교예로서, 상기 성분을 첨가하지 않은 사료 조성물을 사용하였다.As a comparative example, a feed composition without the above ingredients was used.
<실험예 1> 본 발명의 성분들의 메탄 저감 효과 검증<Experimental Example 1> Verification of methane reduction effect of ingredients of the present invention
1. 샘플 준비1. Sample preparation
가. 실험 순서:go. order of experiment:
1) 천연물질을 입자크기 동일하게(1 mm 이하) 분쇄하였다. 1) Natural materials were ground to the same particle size (1 mm or less).
2) DM 측정을 위하여 건조 오븐에서 건조하였다. 2) For DM measurement, it was dried in a drying oven.
3) 'Closed batch culture[McBee, 1953]'을 사용여 천연 후보 물질을 나일론 백에 0.3 g으로 소분하였다. 3) Using 'Closed batch culture [McBee, 1953]', the natural candidate material was divided into 0.3 g in a nylon bag.
나. 실험 재료me. experiment material
- 나일론 백- Nylon bag
- 300 ml 병- 300 ml bottle
다. 실험 장비all. laboratory equipment
- 전자 저울- electronic scale
- 건조 오븐- Drying oven
2. 진탕 인큐베이션2. Shaking incubation
가. 실험 순서: go. order of experiment:
1) 반추위액 준비 1) Preparation of rumen fluid
1-1) 샘플링 전 재료들(나일론 필터, 피펫 50 ml)을 39℃에 셋팅하였다.1-1) Before sampling, the materials (nylon filter, pipette 50 ml) were set at 39°C.
1-2) 캐뉼라 소 2마리의 반추위 방향, 복부 방향(배쪽), 등쪽 방향(꼬리쪽) 총 3부위에서 반추위를 샘플링하였다.1-2) Cannula The rumen of two cows was sampled from three areas: rumen direction, ventral direction (ventral side), and dorsal direction (caudal side).
1-3) 250 ㎛ 나일론 필터를 사용하여 반추위액을 필터링하였다. 1-3) Rumen fluid was filtered using a 250 ㎛ nylon filter.
2) 멘커 완충제(Menke's buffer)[Menke and Steingass, 1988] 제조 2) Manufacture of Menke's buffer [Menke and Steingass, 1988]
3) 반추위액(Rumen Fluid)과 멘컨 완충제를 규정된 비율(1:1-4)로 혼합하였다. 3) Rumen fluid and Mencon buffer were mixed at the specified ratio (1:1-4).
4) CO2를 주입하며 혐기성 상태로 제조하였다.4) CO 2 was injected and manufactured in an anaerobic state.
5) 각 병 당 50 ml씩 반추용액 담은 후, 소분한 병에 Ar 가스를 주입하였다. 5) After putting 50 ml of rumination solution in each bottle, Ar gas was injected into each bottle.
6) 크림퍼로 밀봉한 후에 진탕 인큐베이터(39℃, 120 rpm)에 24시간 발효하였다. 6) After being sealed with a crimper, it was fermented in a shaking incubator (39°C, 120 rpm) for 24 hours.
나. 실험 재료me. experiment material
- 공시 동물의 반추위액 - Ruminal fluid of the tested animal
- McDougall's 완충제 - McDougall's buffer
- 플러싱 가스(Ar 가스, CO2)- Flushing gas (Ar gas, CO 2 )
- Thermos 병 - Thermos bottle
- 20 mn 고무 스톱퍼 - 20 mn rubber stopper
- 알루미늄 시일 - Aluminum seal
- 크림퍼/디캡퍼 - Crimper/Decapper
다. 실험 장비all. laboratory equipment
- 진탕 인큐베이터- Shaking incubator
- 자석 교반기- Magnetic stirrer
3. TGP 측정3. TGP measurement
가. 실험 순서: go. order of experiment:
1) 주사기 바늘을 압력 변환기에 연결하였다. 1) The syringe needle was connected to the pressure transducer.
2) 대기압 영점을 맞춘 후에, 배양병에 바늘을 찔러 넣고 압력을 기록하였다. 2) After setting the atmospheric pressure zero point, a needle was inserted into the culture bottle and the pressure was recorded.
3) 메탄 측정을 위하여 진공 용기에 가스를 주입하였다. 3) Gas was injected into a vacuum vessel to measure methane.
나. 실험 재료me. experiment material
- 24g 시린지 - 24g syringe
다. 실험 장비all. laboratory equipment
- 가스 변환기 - Gas converter
4. pH 측정4. pH measurement
가. 실험 순서: go. order of experiment:
1) 50 ml 튜브에 옮겨 담았다. 1) Transferred to a 50 ml tube.
2) pH 측정 후 기록하였다. 2) pH was measured and recorded.
3) VFA 측정을 위해 원심분리(4℃에서 5분 동안 500× g)를 진행하였다. 3) Centrifugation (500 × g for 5 minutes at 4°C) was performed to measure VFA.
나. 실험 재료me. experiment material
- 50 ml 튜브 - 50 ml tube
다. 실험 장비all. laboratory equipment
- pH 미터 - pH meter
5. 소화율 측정5. Digestibility measurement
가. 실험 순서: go. order of experiment:
1) 필터백 세척한 후, 페이퍼 타월로 물기를 제거하고, 무게를 측정하였다. 1) After washing the filter bag, moisture was removed with a paper towel and the weight was measured.
나. 실험 재료me. experiment material
- 배양병의 나일론 백 - Nylon bag for culture bottle
다. 실험 장비all. laboratory equipment
- 전자 저울 - electronic scale
6. CH6.CH 44 에 대한 가스 샘플링for gas sampling
가. 실험 순서: go. order of experiment:
1) 진공 용기에 담아온 메탄가스 1 ml를 주사기에 담아 크로마토그래피 주입구에 시료를 주입하였다. 1) 1 ml of methane gas contained in a vacuum container was put into a syringe and the sample was injected into the chromatography inlet.
2) 4 내지 5분 후 메탄량, 이산화탄소, 기타 가스 등 측정하여 기록하였다(피크 순서 ETC → CH4 → CO2). 2) After 4 to 5 minutes, the amount of methane, carbon dioxide, and other gases were measured and recorded (peak order ETC → CH4 → CO2).
나. 실험 재료me. experiment material
- 기밀 시린지 - Confidential syringe
- 진공 용기 - Vacuum container
- 표준 가스: He 중 H2 1.0%, CH4 10.1%, CO2 20.1% 및 N2 19.9% - Standard gases: H2 1.0%, CH4 10.1%, CO2 20.1% and N2 19.9% in He.
다. 실험 장비all. laboratory equipment
- 가스 크로마토그래피 - Gas chromatography
▶ 실험결과▶ Experiment results
본 실험예 1은 천연물질(알리신, 아타풀자이트, 미역박)의 제제 조합을 통하여 반추위 발효성상과 이산화탄소, 메탄 배출량에 미치는 영향을 알아보고자 실시하였다. 알리신(A)은 반추위의 소화율과 pH에는 영향을 주지 않았고, 이산화탄소 배출량과 함께 메탄가스를 약 46%까지 저감시킨 것으로 확인할 수 있다(P<0.05). 또한 AB(알리신+아타풀자이트)는 통계적으로 유의미한 변화는 없었으나, 알리신 다음으로 메탄 저감률이 가장 높은 것(23%)으로 확인할 수 있었다.This Experimental Example 1 was conducted to determine the effect on ruminal fermentation characteristics and carbon dioxide and methane emissions by combining preparations of natural substances (allicin, attapulgite, and seaweed meal). Allicin (A) did not affect the digestibility and pH of the rumen, and it was confirmed that it reduced methane gas along with carbon dioxide emissions by about 46% (P<0.05). In addition, AB (allicin + attapulgite) showed no statistically significant change, but was confirmed to have the highest methane reduction rate (23%) after allicin.
따라서, 메탄 발생량은 알리신(4.8 ml) < 알리신+아타풀자이트(6.85 ml) < 알리신+아타풀자이트+미역박(7.93 ml) < 아타풀자이트(8.55 ml) < 아타풀자이트+미역박(9.02 ml) < 알리신+미역박(10 ml) < 미역박(10.96 ml) 순으로 메탄 배출량을 확인할 수 있었다. 알리신(A)과 알리신+아타풀자이트(A+B), 알리신+아타풀자이트+미역박(A+B+C), 아타풀자이트(B) 조합이 대조군보다 메탄량이 감소되었고, 다른 그룹에서는 유의적인 차이를 확인하지 못하였다. 해당 그룹 외의 조합이나 물질의 경우, 메탄을 저감하는 기전보다 과도한 영양성분으로 인해 반추위 내 발효가 증진되어 메탄 배출량이 증가된 것으로 사료된다. Therefore, the amount of methane generated is allicin (4.8 ml) < allicin + attapulgite (6.85 ml) < allicin + attapulgite + seaweed meal (7.93 ml) < attapulgite (8.55 ml) < attapulgite + seaweed meal (9.02 ml) Methane emissions were confirmed in the following order: ml) < allicin + seaweed meal (10 ml) < seaweed meal (10.96 ml). The methane amount was reduced in the combinations of allicin (A), allicin + attapulgite (A + B), allicin + attapulgite + seaweed meal (A + B + C), and attapulgite (B) compared to the control group, while in other groups No significant difference was confirmed. In the case of combinations or substances outside the group, it is believed that methane emissions increased due to enhanced fermentation in the rumen due to excessive nutrients rather than a methane reduction mechanism.
<실험예 2> 천연 메탄저감 첨가제 효능 검정<Experimental Example 2> Efficacy test of natural methane reduction additive
(A)allicin
(A)
(B)attapulgite
(B)
(C)seaweed gourd
(C)
(D)green tea powder
(D)
(E)flaxseed
(E)
상기 표 4에 기재된 성분들을 상기 실험예 1의 방법과 기계를 이용하여 메탄저감 효능을 검정하였다.The ingredients listed in Table 4 were tested for methane reduction efficacy using the method and machine of Experimental Example 1.
▶ 실험결과▶ Experiment results
1. 알리신1. Allicin
시험관내 반추위 발효성상에 따른 pH의 변화에 대한 결과는 상기 표 5와 같다. 반추위 사료의 소화에 적절한 pH의 범위는 6.0 내지 6.8(McCullough et al., 1968)으로 발효 12시간대 이후 모든 첨가제의 pH 범위는 6.55 내지 6.86로 7.5% 첨가량에서 약간 증가되었지만, 대부분 반추위내 적정 pH 범위에 속하는 결과가 나타났다.The results of changes in pH according to in vitro rumen fermentation characteristics are shown in Table 5 above. The appropriate pH range for digestion of rumen feed is 6.0 to 6.8 (McCullough et al., 1968). The pH range of all additives after 12 hours of fermentation was 6.55 to 6.86, slightly increased at the 7.5% addition amount, but mostly within the appropriate pH range in the rumen. Results belonging to were found.
추후 통계적 분석을 통하여, 알리신은 2.5% 첨가량으로 첨가하였을 때, 유의적으로(P<0.05) 메탄 배출량이 감소한 것을 확인할 수 있었고, TGP 비율과 계산해 보았을 때 총 가스 발생량에서 메탄 배출량만 특이적으로 2.5% 첨가량에서 21% 줄은 것을 확인할 수 있다. Through later statistical analysis, When allicin was added at a 2.5% addition amount, it was confirmed that methane emissions were significantly (P<0.05) reduced, and when calculated with the TGP ratio, only methane emissions from the total gas generation decreased by 21% at a 2.5% addition amount. You can check that.
대조군의 CH4 비율(%)과 비교한 결과, 마늘 추출물은 첨가량이 증가할 수록 선형으로 유의적(p < 0.05)으로 감소하는 결과를 나타낸다.As a result of comparing the CH4 ratio (%) of the control group, the garlic extract showed a significant (p < 0.05) linear decrease as the amount added increased.
메탄 배출량은 2.5% 첨가량에서 21% 감소, 7.5% 첨가량에서는 34%까지 감소하였다.Methane emissions decreased by 21% at 2.5% addition amount and by 34% at 7.5% addition amount.
마늘 추출물 성분들 중 하나인 알리신은 메탄생성 단계에 필요한 효소의 활성을 억제하고, 이황화 결합인 S-S(디설파이드기)가 세포질 외막에 있는 단백질과 결합하여, 루멘 박테리아에 영향을 미치지 않고 루멘 메타노 유전자 고균을 특이적으로 억제할 수 있는 잠재력을 가지고 있다.Allicin, one of the components of garlic extract, inhibits the activity of enzymes required for the methanogenesis stage, and the disulfide bond, S-S (disulfide group), binds to proteins in the outer cytoplasmic membrane, thereby preventing lumen methanogene genes without affecting lumen bacteria. It has the potential to specifically inhibit archaea.
2. 아타풀자이트2. Attapulgite
(pH) 시험관내 반추위 발효성상에 따른 pH의 변화에 대한 결과는 상기 표 6과 같다. 반추위 사료의 소화에 적절한 pH의 범위는 6.0 내지 6.8(McCullough et al., 1968)로 발효 12시간대 이후 모든 첨가군의 pH 범위는 6.83까지 약간 증가되었지만, 대부분 반추위내 적정 pH 범위에 속하는 결과가 나타났다.(pH) The results of changes in pH according to in vitro rumen fermentation characteristics are shown in Table 6 above. The appropriate pH range for digestion of rumen feed is 6.0 to 6.8 (McCullough et al., 1968). After 12 hours of fermentation, the pH range of all added groups slightly increased to 6.83, but most results were within the appropriate pH range in the rumen. .
추후 메탄 발생량의 감소 폭이 커 통계분석을 진행하였고, 2.5% 첨가량일 때 메탄 배출량은 3.3% 감소, 7.5 첨가량일 때 8.3% 유의적으로(P<0.05) 감소한 것으로 확인할 수 있다. Later, a statistical analysis was conducted as the decrease in methane emissions was large, and it was confirmed that methane emissions decreased by 3.3% when the amount was added at 2.5% and significantly (P<0.05) by 8.3% when the amount was added at 7.5%.
아타풀자이트 통계적 선형은 유의미한 결과(P<0.05)로, 첨가량이 많아질수록 메탄저감 효과는 증가하는것으로 예측할 수 있다.The statistical linearity of attapulgite is a significant result (P<0.05), and it can be predicted that the methane reduction effect increases as the amount added increases.
4가지 천연 추출물이 시험관내 반추위 발효 성상에 따른 총가스, 이산화 탄소 및 메탄 발생량 결과는 상기 표 7 내지 표 10과 같다. The results of total gas, carbon dioxide, and methane production according to the in vitro rumen fermentation characteristics of the four natural extracts are shown in Tables 7 to 10 above.
(미역박) 미역박은 미역에서 식용 조미료와 알긴산을 추출한 후에 건조한뒤 미역 부산물로 제조한 것으로 버려지는 농산물을 활용하여 첨가제를 제조하였다.(Seaweed meal) Seaweed meal is made from seaweed by-products after extracting edible seasoning and alginic acid from seaweed, drying them, and making additives using discarded agricultural products.
미역박 실험군에서의 메탄 배출량(ml/g DM)은 선형 통계가 유의적으로 감소한것으로 확인되므로 더 많은 첨가량이나 다른 메탄저감제와 복합 시너지 효과를 기대해 볼 수 있다. 또한 IVDMD 소화율이 증가한 것으로 보아 TMR 기초 에너지 사료로서도 활용된 것을 기대해 볼 수 있다(H.G.LEE et al.,2005)Methane emissions (ml/g DM) in the seaweed meal experimental group were confirmed to have significantly decreased linear statistics, so a synergistic effect can be expected with higher addition amounts or with other methane reduction agents. In addition, considering that IVDMD digestibility has increased, it can be expected that it is also used as TMR basic energy feed (H.G.LEE et al., 2005)
(녹차박) 먼저, 녹차박은 메탄을 감소시키는 성분 중 축합형 탄닌 791 mg/kg이 함유 되어있으며 현재 녹차티백과 같이 우리고 난 후에 남은 녹차 부산물을 사용한 것으로 활용도가 매우 높다. (Green tea foil) First, green tea foil contains 791 mg/kg of condensed tannin, one of the ingredients that reduces methane. It is very useful as it is made from green tea by-products left after brewing, like green tea bags.
녹차박 실험군의 메탄 배출량은 7.5% 첨가량에서 4.7%까지 감소된 것을 확인할 수 있다. 총 가스 발생량(TGP)은 줄어들지 않는 것으로 보아 반추위 내 미생물 뿐만 아니라, 총 가스발생량에 영향을 미치지 않고 메탄 발생량이 감소하는 것을 검증하였다.It can be seen that the methane emissions of the green tea cake experimental group were reduced by 4.7% at the 7.5% addition amount. Since the total gas generation (TGP) did not decrease, it was verified that methane generation was reduced without affecting the total gas generation as well as microorganisms in the rumen.
(린씨드) 린씨드는 원물인 씨앗을 갈아서 진행 한것으로, 약간의 메탄 발생량이 감소하였다. 추후 린씨드를 가공 및 제조한 재료로써 재실험할 가능성을 확인 하였다.(Linseed) Linseed is made by grinding the original seed, so the amount of methane produced is slightly reduced. The possibility of re-experimenting with linseed as a processed and manufactured material was confirmed in the future.
(감피) 감피 실험군에서는 IVDMD 소화율이 증가하고 에너지로 쓰이는 TVFA(Total volatile fatty acid)가 약간씩 모두 증가한 것으로 보아 감피가 에너지로 쓰여 생산성이 향상된 것으로 예측할 수 있다. 메탄 배출량은 대조군와 약간의 차이가 있었으나, 유의성은 인정되지 않았다.(Persimmon peel) In the persimmon experimental group, IVDMD digestibility increased and TVFA (Total volatile fatty acid), which is used as energy, both increased slightly, so it can be predicted that productivity improved because persimmon was used as energy. There was a slight difference in methane emissions from the control group, but significance was not recognized.
<실험예 3> 천연 메탄저감 첨가제 농도(첨가량) 검증<Experimental Example 3> Verification of natural methane reduction additive concentration (added amount)
상기 표 11에 기재된 성분들을 상기 실험예 1의 방법 1 내지 5와 기계를 이용하여 메탄저감 효능을 검정하였다.The ingredients listed in Table 11 were tested for methane reduction efficacy using methods 1 to 5 of Experimental Example 1 and a machine.
또한, 하기 실험을 추가로 수행하였다.In addition, the following experiment was additionally performed.
6. 암모니아 질소(NH6. Ammonia nitrogen (NH 33 -N)-N)
가. 실험 순서: go. order of experiment:
1) 샘플 준비1) Sample preparation
① 이전 장에서 준비한 시료를 20,000 g에서 4℃에서 20분간 원심분리하였다. ① The sample prepared in the previous chapter was centrifuged at 20,000 g at 4°C for 20 minutes.
② 상등액 0.5 mL를 1.5 mL 튜브로 운반하였다. ② 0.5 mL of supernatant was transported into a 1.5 mL tube.
2) 표준제제2) Standard formulation
① NH4Cl을 이용하여 NH3-N 표준용액 1 mL를 제조하였다.① Prepare 1 mL of NH 3 -N standard solution using NH 4 Cl.
② 0.2 mL의 HgCl2(wt/vol) 용액과 25%의 메타-인산(wt/vol) 용액을 첨가하였다.② 0.2 mL of HgCl 2 (wt/vol) solution and 25% meta-phosphoric acid (wt/vol) solution were added.
3) NH3-N 분석3) NH 3 -N analysis
① 플레이트 레이아웃을 설계하였다(레이아웃은 샘플 수, 복제 수 및 표준 수까지 변경) ① The plate layout was designed (the layout changes to the number of samples, replications, and standards).
② 2 ㎕의 검체와 표준물을 운반한 후 분석 복제 횟수로 반복하였다. ② After transporting 2 ㎕ of sample and standard, the analysis was repeated with the number of replicates.
③ 알칼리-하이포클로라이트 시약 125 ㎕와 페놀 착색제 147 ㎕를 운반하였다. ③ 125 ㎕ of alkali-hypochlorite reagent and 147 ㎕ of phenol colorant were transported.
⑤ 플레이트를 부드럽게 흔들고 50 내지 57 ℃에서 3 내지 7분간 가열하였다. ⑤ The plate was gently shaken and heated at 50 to 57°C for 3 to 7 minutes.
⑥ 분광 광도계와 소프트웨어를 켜고 파장을 630 nm로 설정하였다. ⑥ Turn on the spectrophotometer and software and set the wavelength to 630 nm.
⑦ 플레이트를 3회 이상 분석하여 측정의 정확도를 높였다. ⑦ The accuracy of measurement was improved by analyzing the plate more than three times.
4) 계산4) Calculation
① 분석간의 평균 OD(nm)를 계산하였다. ① The average OD (nm) between analyzes was calculated.
② 표준 용액의 결과를 바탕으로 선형 회귀 방정식을 적용하였다. ② A linear regression equation was applied based on the results of the standard solution.
③ NH3-N의 농도는 표준곡선에서 계산하며, 분석 복제 사이의 평균 NH3-N을 계산하였다.③ The concentration of NH 3 -N was calculated from the standard curve, and the average NH 3 -N between analysis replicates was calculated.
나. 실험 재료me. experiment material
- HgCl2 1%(wt/vol) 및 25% 메타-인산(wt/vol) 용액 - HgCl2 1% (wt/vol) and 25% meta-phosphoric acid (wt/vol) solution
- NH4Cl -NH4Cl
- NaOH -NaOH
- NaOCl -NaOCl
- Pheno -Pheno
- 나트륨 - 니트로페리사이나이드 - Sodium - Nitrophericinide
- 알칼리-하이포클로라이트 시약 - Alkali-hypochlorite reagent
- 페놀 칼라 - 시약 - Phenol Color - Reagent
다. 실험 장비all. laboratory equipment
- 가스 크로마토그래피 - Gas chromatography
- 분광법 - Spectroscopy
- 96-웰 플레이트 - 96-well plate
7. 휘발성 지방산(VFA)7. Volatile fatty acids (VFA)
가. 실험 순서: go. order of experiment:
1) 샘플 준비 1) Sample preparation
① 샘플을 4℃에서 20분 동안 20,000g에서 원심분리하였다. ① The sample was centrifuged at 20,000g for 20 minutes at 4°C.
② 상등액 0.5 mL를 1.5 mL 튜브에 옮겼다. ② Transfer 0.5 mL of supernatant to a 1.5 mL tube.
2) 표준물 제조 2) Standard preparation
① NH4Cl을 사용하여 NH3-N 표준용액 1 mL 제조하였다.① 1 mL of NH 3 -N standard solution was prepared using NH 4 Cl.
② 0.2 mL HgCl2(wt/vol)와 25% 메타-인산(wt/vol)을 표준 용액에 추가하였다.② 0.2 mL HgCl 2 (wt/vol) and 25% meta-phosphoric acid (wt/vol) were added to the standard solution.
나. 실험 재료me. experiment material
- 오박색 와이드 개방 스크류 톱(write-on spot) -Orange wide opening screw top (write-on spot)
- 블루 캡, PRFE/Red 실리콘 고무 셉텀 - Blue cap, PRFE/Red silicone rubber septum
- GC 표준 시린지 - GC standard syringe
- 울트라-불활성 라이너 - Ultra-inert liner
- 그린 셉텀 - Green Septum
- 2% 피발산 - 2% pivalic acid
- 표준 VFA 용액(wt/vol) - Standard VFA solution (wt/vol)
다. 실험 장비all. laboratory equipment
- 원심분리기 - Centrifuge
- 가스 크로마토그래피 - Gas chromatography
(pH) 시험관내 반추위 발효성상에 따른 pH의 변화에 대한 결과는 표 12와 같다. 반추위 섬유질 함량이 높은 사료의 소화에 적절한 pH의 범위는 6.0 내지 6.8(McCullough et al., 1968)로 발효 12시간대 이후 모든 첨가군의 pH 범위는 6.55 내지 6.73로 2.5% 첨가량에 약간 증가되었지만, 대부분 반추위내 적정 pH 범위에 속하는 결과가 나타났다.(pH) The results of changes in pH according to in vitro rumen fermentation characteristics are shown in Table 12. The appropriate pH range for digestion of feed with high ruminal fiber content is 6.0 to 6.8 (McCullough et al., 1968). After 12 hours of fermentation, the pH range for all groups was 6.55 to 6.73, slightly increasing at the 2.5% addition amount, but most The results were found to be within the appropriate pH range in the rumen.
알리신 2.5% 첨가량에서 메탄 배출량(mL/g DM)이 9.2% 감소한 것을 확인할 수 있었다. 2.5%부터 메탄이 유의적(P<0.05)으로 줄기 시작하는 것을 검증하였고, 통계적으로 선형의 유의미한 수치를 보아(p=0.01), 첨가량이 높아질수록 메탄 배출량은 감소할 것으로 예측된다.It was confirmed that methane emissions (mL/g DM) decreased by 9.2% at the addition amount of 2.5% allicin. It was verified that methane begins to decrease significantly (P<0.05) from 2.5%, and given the statistically significant linear value (p=0.01), it is predicted that methane emissions will decrease as the amount added increases.
총 가스 생산량(TGP)는 변화에 없으며 TVFA(mM)생산성에는 특별한 변화가 없는 것을 확인하였다. 이는 특별하게 메타노젠 미생물만 타겟팅하여 메탄을 감소시키거나 메탄발생에 관여하는 효소만 억제한다는 의미로 반추위내 발효성상에는 영향을 미치지 않고, 메탄만을 감소시키는 고무적인 결과이다.It was confirmed that there was no change in total gas production (TGP) and no particular change in TVFA (mM) productivity. This means that it specifically targets only methanogenic microorganisms to reduce methane or inhibits only the enzymes involved in methane production, which is an encouraging result as it only reduces methane without affecting the fermentation characteristics in the rumen.
(IVDMD) 건물 소화율의 지표로써 대조군에 비하여 모든 실험군에서 IVDMD가 유의적으로 높게 나타났다(P<0.05)(IVDMD) As an indicator of dry matter digestibility, IVDMD was significantly higher in all experimental groups compared to the control group (P<0.05)
(CH4) 총가스 발생량(TGP)에 대비하여 메탄 배출량만 특이적으로 2.5%에서 유의적(P<0.05)으로 감소하였으며, 아타풀자이트의 첨가량이 증가 할 수록 메탄 발생량이 유의적으로 선형으로 감소하였다(P<0.05).(CH4) Compared to total gas generation (TGP), methane emissions specifically decreased significantly (P<0.05) from 2.5%, and as the amount of attapulgite added increased, methane emissions significantly decreased linearly. (P<0.05).
(TVFA) 아타풀자이트의 시험관내 반추위 발효 시간에 따른 TVFA 결과는 표 13과 같다. 발효 12시간대에 아타풀자이트는 대조군에 비해 모든 실험군에서 유의적(P<0.05)으로 높아진 것을 확인할 수 있다. 아세테이트 농도는 감소하고 프로피오네이트 농도가 증가하는 것으로 보아 프로피오네이트 생성에 수소가 쓰여 메탄 합성에 필요한 수소가 줄어들게 되어 메탄 생성이 감소하게 된 것으로 추측해 볼 수 있다.(TVFA) The TVFA results according to the in vitro rumen fermentation time of attapulgite are shown in Table 13. It can be seen that attapulgite increased significantly (P<0.05) in all experimental groups compared to the control group at 12 hours of fermentation. As the acetate concentration decreases and the propionate concentration increases, it can be assumed that hydrogen is used to produce propionate, and the hydrogen required for methane synthesis decreases, resulting in a decrease in methane production.
<실험예 4> 본 발명의 메탄 저감 첨가제를 급여한 반추 동물의 메탄 발생량 변화(생체내) 검증<Experimental Example 4> Verification of changes in methane production (in vivo) in ruminants fed with the methane reduction additive of the present invention
1. 실험 설계(공시축 및 사양 관리)1. Experimental design (simultaneous axis and specification management)
연구 절차 및 공시축의 사양관리는 홀스타인(380±20kg) 15~16개월령의 10두를 공시 하였고, 공시축은 하루에 2회 농후사료 6 kg, 조사료 5 kg를 10:00h와 17:00h 나누어서 주었으며, 물은 자유 섭취하도록 하였다. 실험 기간은 2022년 10월-11월 총 5주로 적응기간 1주, 실험기간 4주간 복합 사료첨가제(마늘추출물, 아타풀자이트)를 급여하였다. 사료첨가제는 앞서 실험을 바탕으로, 농후사료 기준, 마늘추출물 2.5% 아타풀자이트 5.5% 조합 0.48 g을 주었으며 농후사료에 탑드레싱하여 10:00h에 급여하였다. 체중과 메탄 발생량(0주차)을 측정하여 개체별 그룹화를 진행하였다. The research procedure and feeding management of the test animals were conducted on 10 Holstein (380±20 kg) 15-16 month old cows, and the test animals were given 6 kg of concentrated feed and 5 kg of roughage twice a day at 10:00h and 17:00h. Water was allowed to be consumed freely. The experiment period was a total of 5 weeks from October to November 2022, with 1 week of adaptation period and 4 weeks of experiment period where complex feed additives (garlic extract, attapulgite) were fed. As for the feed additive, based on previous experiments, 0.48 g of a combination of 2.5% garlic extract and 5.5% attapulgite was given based on the concentrated feed, and the concentrated feed was top-dressed and fed at 10:00 h. Body weight and methane production (week 0) were measured and grouped by individual.
2. 사료성분 및 NRC 요구량 분석2. Analysis of feed ingredients and NRC requirements
실험에 사용된 마늘추출물(알리신)과 아타풀자이트는 (주)가야바이오의 제품(Green up)을 제조하여 사용하였다.Garlic extract (allicin) and attapulgite used in the experiment were manufactured by Gaya Bio Co., Ltd. (Green up).
현재 실험에서 조사료와 농후사료의 성분 분석을 하여 계산한 결과, DMI 9.7, TDN 65.08%, CP 9.64%으로 NRC 기준으로 영양소 요구량을 비교해 보았다. 실험 착수 체중이 최저는 344 kg, 최고는 420 kg이였으므로 NRC의 일일 영양소 요구량 BW 400 kg 기준으로 ADG 0.5 내지 1.1 kg/일 비교했을 때 TDN 일일 총 섭취량은 ADG 1.1 kg 기준으로도 부족하지 않은 것을 확인하였다. 조단백질은 ADG 0.8 kg/일 기준으로 CP 함량이 조금 낮은 수치를 보였으나 요구량에는 -0.05 kg이 부족한 것으로 거의 충족했다고 볼 수 있다. In the current experiment, as a result of analyzing the composition of roughage and concentrated feed, the nutrient requirements were compared based on the NRC with DMI 9.7, TDN 65.08%, and CP 9.64%. The lowest body weight at the start of the experiment was 344 kg and the highest was 420 kg, so when comparing 0.5 to 1.1 kg/day of ADG based on the NRC's daily nutrient requirement BW of 400 kg, the total daily intake of TDN was not insufficient even at 1.1 kg of ADG. Confirmed. Crude protein showed a slightly lower CP content based on ADG of 0.8 kg/day, but it can be considered that the requirement was almost met, falling short by -0.05 kg.
3. 메탄 측정기기3. Methane measuring instrument
메탄 측정기기는 반도체 레이저를 이용한 근적외선 흡수 분광법에 의하여 공기중에 메탄을 검지하는 기기 레이저 메탄 검출기(LMD, mini-G(50A), Tokyo Gas Engineering Co., Japan)(도 3a)를 이용하는 LMD법을 활용하였다. 공시축들을 스타치온에 고정한 후 LMD를 이용하여 메탄 배출 농도를 측정하였다. 공시동물과 1 M 떨어진 거리에서 LMD에서 발사되는 레이저가 동물의 코 아래 부분에 위치하도록 하고, 반사되어 돌아오도록 하였다. The methane measuring device is the LMD method using a laser methane detector (LMD, mini-G (50A), Tokyo Gas Engineering Co., Japan) (Figure 3a), which detects methane in the air by near-infrared absorption spectroscopy using a semiconductor laser. It was utilized. After fixing the blank axes to the starchion, methane emission concentration was measured using LMD. The laser emitted from the LMD at a distance of 1 m from the test animal was positioned below the animal's nose and reflected back.
메탄 배출 농도는 실험 개시 1 주일 전, 개시 1,2,4 주일 후 총 2일간 오전사료 급여 기준으로(급여±1, 2 h) 하루 총 4회 매 0.5초 간격으로 6분간 측정하였다. 6분간 측정한 이유는 트름이 주로 메탄가스를 방출하는 주요 루트이며 1 내지 3분씩 불규칙적으로 발생하므로, 6분간 측정하여 최소 한번의 트름 피크를 포함하여 측정할 수 있다(도 3b). 측정한 데이터가 0이거나, 1,000 ppm-m 을 초과한 경우에는 데이터에서 제외하였으며 이러한 농도를 기준으로 AMPD(automatic multi-scale peak detection) 알고리즘 패키지(R 소프트웨어 버젼 4.0.2)를 사용하여 하루의 호흡과 트름 간의 평균 메탄 배출량을 산출하여 계산하였다. 산출된 메탄 발생량을 DMI에 따른 메탄 발생량과 ADG에 따른 메탄 발생량으로 표기하였다. Methane emission concentration was measured 4 times a day for 6 minutes at intervals of 0.5 seconds, based on morning feed feeding (±1, 2 h) for a total of 2 days, 1 week before the start of the experiment and 1, 2, and 4 weeks after the start of the experiment. The reason for measuring for 6 minutes is that burps are the main route that emits methane gas and occur irregularly every 1 to 3 minutes, so by measuring for 6 minutes, measurements can be made including at least one burp peak (Figure 3b). If the measured data was 0 or exceeded 1,000 ppm-m, it was excluded from the data, and based on this concentration, daily respiration was calculated using the AMPD (automatic multi-scale peak detection) algorithm package (R software version 4.0.2). It was calculated by calculating the average methane emissions between and. The calculated methane production amount was expressed as methane production amount according to DMI and methane production amount according to ADG.
4. 검사 항목 및 통계 분석4. Test items and statistical analysis
(건물 섭취량 및 체중 측정) 건물 섭취량은 실험 기간동안 측정하였으며 배합사료는 급여 30분 후에 잔여량을 측정하였다. 체중은 실험 개시 전, 개시 후 4주 뒤에 측정하였다.(Dry matter intake and body weight measurement) Dry matter intake was measured during the experiment period, and the remaining amount of the mixed feed was measured 30 minutes after feeding. Body weight was measured before and 4 weeks after the start of the experiment.
검사 항목으로 체중과 일일섭취량, 일일증가량, 혈액의 CBC, MPT를 보고자 한다. CBC는 빈혈이나 감염, 혈액 응고능력이나 면역계 반응 능력에 대한 정보를 제공하며 MPT는 대사 판정 지표로 혈중의 대사 산물의 농도를 측정하여 대사작용 이상여부를 판단할 수 있다. 마늘 추출물에는 간에서 합성되는 글루타티온 생산에 필수적인 황 함유 아미노산이 다량 함유되어 항산화 효과에 매우 좋다는 것으로 알려져있다. Test items include body weight, daily intake, daily gain, blood CBC, and MPT. CBC provides information on anemia, infection, blood coagulation ability, and immune system response ability, and MPT is a metabolic judgment index that measures the concentration of metabolites in the blood to determine whether metabolic activity is abnormal. Garlic extract is known to contain a large amount of sulfur-containing amino acids, which are essential for the production of glutathione synthesized in the liver, and is very good for antioxidant effects.
본 연구는 Statistical Analysis Syste(version 9.4)의 PROC MIXED와 T-TEST에 따라 처리되었다. 통계적 유의성은 5% 유의수준으로 평가하였고, 5% 내지 10% 사이인 경우에 경향성이 있음으로 평가하였다. This study was processed according to PROC MIXED and T-TEST of the Statistical Analysis System (version 9.4). Statistical significance was evaluated at a 5% significance level, and a tendency was evaluated when it was between 5% and 10%.
5. MPT5.MPT
(GOT, GPT) 농도는 간기능 검사에 이용되는 효소의 일종으로 간세포가 파괴되거나 손상되면 밖으로 GGT 및 AST가 유리되게 되며 이 효소들은 에너지 섭취량, 스트레스 및 질병등의 영향을 받아 증가하게 되나, 유의미한 차이가 없었던 것으로 간기능에 이상이 없었던 것으로 판단된다.(GOT, GPT) concentration is a type of enzyme used in liver function tests. When liver cells are destroyed or damaged, GGT and AST are released. These enzymes increase under the influence of energy intake, stress, and disease, but there is no significant difference. Since there was no difference, it is judged that there was no abnormality in liver function.
(BUN) 수치 감소는 단백질 섭취 부족, 반추위 분해 단백질부족(반추위 미생물 성장감소, 호르몬 생성저하 등)으로 볼 수 있으나 총 단백질은 증가하였으며, 통계적으로 유의미한 차이가 없었으므로 이상이 없는 것으로 판단된다A decrease in (BUN) levels can be seen as a lack of protein intake or lack of rumen decomposed protein (reduced rumen microbial growth, decreased hormone production, etc.), but total protein increased, and there was no statistically significant difference, so it is judged that there is no problem.
(TCHO) 주로 회장에서 다른 물질과 함께 흡수되는데, 주요 스테로이드 호르몬의 준구체, 담즙산의 전구체, 세포막 필수 구성성분으로 대사적 기능을 수행하는 역할을 하며, 종합적인 영양 상태와 간 기능을 나타내며 간 기능이 저하되거나 영양소의 섭취부족 시 낮아지고, 지방질 섭취량은 과다시에 높아지게 되나, 유의미한 차이가 없었던 것으로 간 기능에 이상이 없었던 것으로 판단된다.(TCHO) is mainly absorbed along with other substances in the ileum. It plays a role in performing metabolic functions as a quasi-sphere of major steroid hormones, a precursor of bile acids, and an essential component of cell membranes. It indicates the overall nutritional status and liver function. It decreases when nutrient intake is insufficient or increases when fat intake is excessive, but there was no significant difference, indicating that there was no abnormality in liver function.
(글루코스) 인슐린 수치의 증가로 인한 저혈당 효과를 볼 수 있으나 대조군와 첨가군 간의 차이가 없다.(Glucose) A hypoglycemic effect can be seen due to an increase in insulin levels, but there is no difference between the control group and the added group.
이로써, 대사이상 지표들은 대조구와 처리구간의 차이가 없었으므로 대사에는 문제가 없었던 것으로 판단할 수 있다.As a result, there was no difference in metabolic abnormality indicators between the control and treatment groups, so it can be determined that there was no problem with metabolism.
6. 메탄 농도 분석6. Methane concentration analysis
동물이 배출하는 메탄농도는 섭취량과 일일증체량 차이로 인하여 메탄 발생량을 보정하여 표현 하였다. 각 동물의 메탄 농도(ppm-m/kg DMI)는 LMD로 측정한 수치(ppm-m)을 평균 건물섭취량으로 나눈 건물 섭취량당 메탄 농도를 의미한다. 또한 동물의 메탄 농도(ppm-m/kg ADG)는 LMD로 측정한 수치(ppm-m)을 평균 일일증체량으로 나눈 일일증체량당 메탄 농도를 의미한다. The concentration of methane emitted by the animal was expressed by correcting the amount of methane produced due to the difference in intake and daily weight gain. The methane concentration (ppm-m/kg DMI) of each animal refers to the methane concentration per dry matter intake divided by the value measured by LMD (ppm-m) by the average dry matter intake. Additionally, the animal's methane concentration (ppm-m/kg ADG) refers to the methane concentration per daily body weight divided by the value measured by LMD (ppm-m) by the average daily weight gain.
반추위내 메탄 발생량 중 호흡과 트름을 분리하여 계산하였을 때 첨가제 처리군과 대조군의 차이는 통계적으로 유의미하였다(P<0.05) 이는 아타풀자이트와 알리신의 복합 첨가제는 항메타노젠과 메탄 발생 효소를 저해한다는것으로 해석할 수 있다. When the amount of methane produced in the rumen was calculated separately from respiration and belching, the difference between the additive treatment group and the control group was statistically significant (P<0.05). This means that the combined additive of attapulgite and allicin inhibits anti-methanogen and methanogenic enzymes. It can be interpreted as doing.
트름의 메탄 발생량은(ppm/m) 1주차 40.79%> 2주차 32.80%> 3주차 45.83%, 호흡의 메탄 발생량은 1주차 13.82%> 2주차 39.62%> 3주차 57.72% 로 감소한 것을 검증하였다(도 4 및 표 19 참조). It was verified that the amount of methane produced by burping (ppm/m) decreased from 40.79% in the 1st week > 32.80% in the 2nd week > 45.83% in the 3rd week, and the amount of methane produced in respiration decreased from 13.82% in the 1st week > 39.62% in the 2nd week > 57.72% in the 3rd week ( (see Figure 4 and Table 19).
메탄 발생량을 시간에 흐름에 따라, 호흡은 1주차에는 메탄감소의 경향성이 있으며 2주차부터 유의미적으로 감소되는것을 확인할 수 있었고(p<0.05), 트름은 1주차부터 감소하여 급여하는 기간이 늘어날수록 메탄 저감 효과가 더욱 큰 것을 확인할 수 있었다(p<0.001). 즉 대조군에 비하여 모든 처리군은 메탄 생성량이 감소하는 결과를 우수한 결과를 보였다. Depending on the amount of methane production over time, respiration tended to decrease in methane in the first week and significantly decreased from the second week (p<0.05), and burping decreased from the first week, increasing the feeding period. It was confirmed that the methane reduction effect was greater (p<0.001). In other words, compared to the control group, all treatment groups showed excellent results in reducing methane production.
본 발명에서는 천연 추출물에서 유래한 반추위 메탄 저감용 사료첨가제 개발을 위하여 여러가지 천연물질의 조합과 첨가량 실험을 체계적으로 실시하였다. 결과적으로 마늘추출물과 아타풀자이트의 복합 사료첨가제가 메탄 발생량 저감에 긍정적인 결과를 보였으므로, 알리신의 2.5% 첨가량과 아타풀자이트의 5.5% 첨가량은 메탄 생성량을 저감 시킬 수있는 사료첨가제의 가능성을 제시하였다.In the present invention, combinations and addition amounts of various natural substances were systematically tested to develop a feed additive for reducing rumen methane derived from natural extracts. As a result, the composite feed additive of garlic extract and attapulgite showed positive results in reducing methane production, so the addition of 2.5% of allicin and 5.5% of attapulgite showed the potential as a feed additive that can reduce methane production. presented.
따라서, 본 발명에서는 반추위 미생물 발효와 사료 이용 효율, 대사에 대한 부정적인 효과 없이, 복합 천연 추출물의 선정과 적정한 투여량 비율을 통하여 메탄 발생량 감소의 효과를 검증하였다. Therefore, in the present invention, the effect of reducing methane production was verified through the selection of complex natural extracts and appropriate dosage ratio without negative effects on rumen microbial fermentation, feed use efficiency, and metabolism.
Claims (10)
상기 알리신:아타풀자이트:미역박 및 린씨드의 중량비는 0.5 내지 2.5:2.5 내지 5.5:0.5 내지 5.5인, 반추위 내 메탄가스 저감을 위한 사료 첨가제.Contains allicin, attapulgite and seaweed meal and linseed,
The weight ratio of allicin: attapulgite: seaweed meal and linseed is 0.5 to 2.5:2.5 to 5.5:0.5 to 5.5, a feed additive for reducing methane gas in the rumen.
사료 조성물 100 중량%를 기준으로
상기 알리신의 양은 0.5 내지 2.5 중량%이며,
상기 아타풀자이트의 양은 2.5 내지 5.5 중량%이며,
상기 미역박과 린씨드의 합한 양은 0.5 내지 5.5 중량%인 반추위 내 메탄가스 저감을 위한 사료 첨가제를 포함하는,
반추 동물용 사료 조성물.Contains allicin, attapulgite and seaweed meal and linseed,
Based on 100% by weight of feed composition
The amount of allicin is 0.5 to 2.5% by weight,
The amount of attapulgite is 2.5 to 5.5% by weight,
The combined amount of the seaweed meal and linseed contains a feed additive for reducing methane gas in the rumen of 0.5 to 5.5% by weight,
Feed composition for ruminants.
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