201136528 六、發明說明： 【發明所屬之技術領域】 本發明係關於一種具有反g胃之動物（以下稱為「反芻動 物」）之飼料利用效率及乳量改善。更詳言之，係關於一種 反芻動物之飼料利用效率改善劑，其在牛、綿羊、山羊等之 反芻動物中，改善一般所謂瘤胃醱酵的第一胃内醱酵（以下 亦稱為「瘤胃醱酵」），且增加體内之有用菌，提升飼料消 化率並減少能量損失，改善飼料利用效率，結果發揮可飼育 出改善了乳量的健康家畜。 尚且，本發明中所謂飼料消化率，通常係指DM(DRY MATTER)消化率，具體而言，係以飼料消化率（％)=「1 —(糞 固形物重量)/(飼料固形物重量）」xlOO所表示者。此DM消 化率並未反映出飼料成分中被分解而以氣體所排出的成 分、或排出至尿中的成分等(耗損成分）。本發明中，係將亦 包括了此等耗損成分（能量損失）的整體效率視為飼料利用 效率。 【先前技術】 反芻動物因為具有與豬等單胃動物不同的特殊消化吸收 機構與能量代謝機構，故其飼育、管理時需要特別的知識或 技術。 而且在牛、綿羊、山羊等之乳亦成為商品的反芻動物中， 身體狀況不良或疾症等造成的乳量降低，對酪農農家而言為 100108238 4 201136528 嚴重的經濟問題。又，即使在動物的身體狀況良好的情況， 若增加每一頭的乳量，則連帶地增加酪農家的收入，故較 佳。再者，此種效果最好能由較少的飼料而效率佳地獲得。 為知在反芻動物的情況，其第一胃、亦即瘤胃中的微生 物（細菌與原生動物(Protozoa，稱為原生動物或原蟲），·以下 亦稱為「瘤胃内微生物」）由於產生必須胺基酸，故一般認 為並無於飼料中補充必須胺基酸的必要性。然而，近年來開 始認為為了高乳量的穩定生產或維持健康而必須補充一定 里的蛋白質或必須胺基酸。例如，於乳牛中，雖嘗試了添加 各種營養素與屬於蛋白質源的榖類或魚粉或血粉等作為飼 料添加物，但此等有牛本身的喜好及此等蛋白質源之攝取造 成牛代謝負擔的問題，無法得到充分效果。再者，作為增大 乳量的方法，已研討有在飼料中添加於瘤胃内不被分解之保 邊胺基酸(專利讀2) ’但添加時麟調節等複雜，保護胺 基酸之吸收差而效果亦不足。其他已知有對反籍動物經口投 予含有胺級之液狀&成物(專散獻丨），但錄狀組成物 中之胺基酸的分解、投予方法、飼養方法等出現問題。 另外’已知藉由對飼育動物投予枯草桿菌，將抑制飼育動 物體内之病雜細g的増殖，並增加該減之體重、以及改 善飼料要求率’例如’已知以枯草桿菌(Bacillus SU議S)C_3102株之生菌體作為有效成分的幼豬用調配铜料 造成增體效果等(專利文獻3)，再者，以枯草桿菌JA-ZK株 100108238 5 201136528 作為有效成分的飼料添加劑，可得軸育動物體内之 細菌增殖抑制賴動物之㈣増加、及喃要轉的改盖效 果（專利文獻4);已知使用以枯草桿菌= sub卿C_31G2株或含枯草桿菌之枯草®则株作為有1 成分的抑·，可抑制主要存在於鳥類體内之沙Η桿菌等有 害細菌之繁殖的方法(專利文獻5、6)等。然而，任一種= 具體揭不㈣制作為具有與豬或鳥料同之特異消化吸 收機構之反_物的匈料利用效率改善劑或乳量改善劑。 對於牛等反®動物，雖嘗試過給予經醋酵之飼料或容易醋 酵之飼料以提高飼料消化，但若—次大量給予容㈣酵之飼 料等m内容易發生急遽醱酵，使瘤胃内環境酸性 化’微生物造細酵料降低或引起異常而發生大量氣體， 失增大。·^—時性的瘤胃酸中毒的發生亦使食慾 降低。而且，此種現象弓1起第一胃（瘤胃）胃液的pH異常、 二壁&傷 '胺類等之異常代謝產物的產生、異常之醱酵熱 等，而妨礙正f的消化或吸收。再者，瘤胃内微生物以外之 、田菌的異吊增殖、其移行至血中而成為消化道疾病、肝障 礙、運動器疾病、繁殖障礙、熱射病、乳房炎、皮膚炎等之 原因’造成問題。 等反离動物的特異消化吸收機構或能量代謝機構 為周知技術=項(料射獻υ，於«料明其特徵。 猪焉等單月動物中’所攝取的飼料在以口腔、食道、胃、 100108238 6 201136528 小腸、大腸_序被輸送時，_本身所分泌的酵素被依序 分解(例如殿粉等成為葡萄糖），而由消化管壁所吸收。包括 人類在内的豬等單胃動物因為需要糖作為能量源，而被稱為 糖動物。 另一方面’牛等反离動物具有與單胃動物不同之消化•吸 收構成。第一胃内的酸酵稱為瘤胃酸酵，其特徵在於由植物 纖維等碳水化合物心成揮發㈣㈣(VF顺再 =:4二1粗倒料再次回到口中而再°且嚼㈣，並再 :人進盯第-月的瘤胃醱酵。第一胃被稱為所謂 槽」’棲息者各式各樣的厭氣性微生物 物 =:r她，娜彳 Γ。，等_由_直接 此外，並成為貢_組織所消耗 一+的活力源。揮發性_酸(视)中，能 = 丙酸。而且，牛望千取间者為 肪酸動物。 動物被稱為㈣肪酸取得能量源的脂 胃=乎3蛋白質’在豬等單胃動物中，飼料之蛋白質在 的消化酸去 運至腸中’於腸中藉由動物本身所具有 、的作用而分解為胺基酸後再 '、 等反-物中’係藉由瘤胃内微生物之蛋白二 100108238 F ™ 201136528 而分解為胺基酸、接著分解為氨，再者，微生物以該氨作為 材料而重新合成出必須胺基酸、亦即高營養價值的微生物體 蛋白質（菌體蛋白質、轰體蛋白質）。如此形成的「微生物體 蛋白質」係與胃内容物一起運送至第4胃之後，與單胃動物 同樣地進行主要於小腸内被分解為胺基酸而被吸收的過 程。於此，牛等蛋白質的消化•吸收尚有另一個路徑，在瘤 胃内不被微生物分解而直接運送至小腸，於其中被消化的過 瘤胃蛋白質（bypassprotein)。此係與微生物體蛋白質一起被 稱為代謝蛋白質，但此等係於小腸中成為胺基酸而吸收。 士 0 4匕，嚷 乐〜胃（瘤胃）為所謂的活的醱酵裝置，供給必要的 能量源，另〜+ . 乃方面，以醱酵副產物之型式生成出甲烷或碳酸 氣體、氮及小θ y 於 %久>1氧、氩、硫化氫等，其幾乎不被利用而以氣 體排出至大氣中，成為能量損失。又，在第-胃中之醱酵不 足夺飼料中之養分成為糞中之氮等有機物或礦物質類等無 機物’未被利用而排出。再者，一般認為由飼料所攝取的氮 之三分之^ 一 〜成為糞，三分之一排出至尿中，剩餘則移至乳中 或蓄積於體内。 k而’孔量增大之健康家畜的飼育時，重要的是改善瘤胃 Μ月Μ生物之增加或活性化、高能量效率之丙酸產生 的提升或向營養價值之蛋白源的微生物體蛋白質合成的提 升飼料消化率之提升及能量損失減低、提高飼料利用效 率而要求開發出此種有效之反努動物的飼料利用效率改善 100108238 8 201136528201136528 VI. Description of the Invention: [Technical Field of the Invention] The present invention relates to an improvement in feed utilization efficiency and milk amount of an animal having an anti-g stomach (hereinafter referred to as "anti-mite"). More specifically, it relates to a feed utilization efficiency improving agent for ruminants, which improves the first intragastric fermentation of the so-called rumen fermentation in ruminants such as cows, sheep, and goats (hereinafter also referred to as "rumen" Fermentation"), and increase the useful bacteria in the body, improve feed digestibility and reduce energy loss, improve feed utilization efficiency, and as a result, can produce healthy livestock that can improve the amount of milk. Further, the so-called feed digestibility in the present invention generally means DM (DRY MATTER) digestibility, specifically, feed digestibility (%) = "1 - (fecal solid weight) / (feed solid weight) "xlOO said. This DM digestion rate does not reflect the components of the feed component that are decomposed and discharged by the gas, or the components that are discharged into the urine (depletion components). In the present invention, the overall efficiency including such lossy components (energy loss) is regarded as feed utilization efficiency. [Prior Art] Since ruminants have special digestion and absorption mechanisms and energy metabolism mechanisms different from monogastric animals such as pigs, they require special knowledge or techniques for breeding and management. In addition, in the ruminants where cows, sheep, goats, etc., are also commodities, the amount of milk caused by poor physical condition or illness is reduced, which is a serious economic problem for the farmer's farmer 100108238 4 201136528. Moreover, even if the animal's physical condition is good, it is better to increase the amount of milk per head, which increases the income of the farmer's family. Moreover, such an effect is preferably obtained efficiently with less feed. In order to know the situation of ruminants, the first stomach, that is, the microorganisms in the rumen (bacteria and protozoa (protozoa), hereinafter referred to as "ruminal microorganisms") Amino acids are generally considered to be unnecessary for the supplementation of essential amino acids in the feed. However, in recent years, it has been thought that a certain amount of protein or essential amino acid must be added for stable production or maintenance of high milk content. For example, in dairy cows, although various nutrients and scorpions or fishmeal or blood meal belonging to the protein source have been tried as feed additives, the preference of the cattle itself and the intake of these protein sources cause the metabolic burden of the cattle. Can't get enough results. Furthermore, as a method of increasing the amount of milk, it has been studied to preserve the marginal amino acid which is not decomposed in the rumen in the feed (patent read 2), but the addition of the lining adjustment is complicated, and the absorption of the amino acid is protected. Poor and insufficient results. Others are known to administer a liquid-like & ampoule containing an amine grade to a reciprocal animal, but the decomposition, administration method, and feeding method of the amino acid in the recorded composition appear. problem. In addition, it is known that by administering Bacillus subtilis to a breeding animal, the cultivation of the disease m in the breeding animal is inhibited, and the weight loss is increased, and the feed requirement rate is improved, for example, 'Bacillus subtilis is known. SU) S) The growth-promoting effect of the raw material of the C_3102 strain as an active ingredient in the preparation of copper material for young pigs (Patent Document 3), and further, the feed additive of Bacillus subtilis JA-ZK strain 100108238 5 201136528 as an active ingredient It is possible to obtain the effect of the bacterial growth inhibition in the abalone animal (4), and the effect of the change of the turmeric (Patent Document 4); it is known to use Bacillus subtilis = subqing C_31G2 strain or subtilis® containing Bacillus subtilis In the case of the strain, it is possible to suppress the growth of harmful bacteria such as Salmonella, which is mainly present in birds, (Patent Documents 5 and 6). However, any one of them is specifically produced as a reaction efficiency improving agent or a milk amount improving agent having a counter-product of a specific digestion and absorption mechanism similar to that of pigs or birds. For cows and other anti-® animals, although they have tried to give the vinegar-fed feed or the vinegar-feeding feed to improve the digestion of the feed, if the feed is given in a large amount, the feed is easy to occur in the rumen. The acidification of the environment 'microbial fine yeasts reduce or cause abnormalities and a large amount of gas occurs, and the loss increases. • The occurrence of rumen rumen acidosis also reduces appetite. Moreover, this phenomenon involves the abnormal pH of the first stomach (rumen) gastric juice, the production of abnormal metabolites such as the second wall & 'amine', and the abnormal heat of fermentation, which hinders the digestion or absorption of positive f. . Furthermore, the growth of the bacterium in the rumen other than the microbes in the rumen, which migrates to the blood and becomes a cause of digestive tract diseases, liver disorders, motor disease, reproductive disorders, heat shock, mastitis, dermatitis, etc. Causes problems. The specific digestion and absorption mechanism or energy metabolism mechanism of the reciprocal animal is known as the technology = item (the material is given, in the characteristics of the material. The feed in the month of the pig, etc.) is in the mouth, the esophagus, the stomach. 100108238 6 201136528 When the small intestine and large intestine are transported, the enzyme secreted by _ itself is decomposed sequentially (for example, the powder into glucose), and is absorbed by the wall of the digestive tract. Single stomach animals such as humans Because sugar is needed as an energy source, it is called a sugar animal. On the other hand, 'negative animals such as cattle have different digestion and absorption composition than monogastric animals. The acid yeast in the first stomach is called rumen acid yeast, and its characteristics. It is caused by the carbohydrates such as plant fibers to volatilize (4) (4) (VF shun == 4 2 1 rough dumping back to the mouth again and then chew (four), and again: people into the first month of rumen fermentation. First The stomach is called the so-called trough"'s inhabitants' various anaerobic microbial substances =: r she, Na 彳Γ., etc. _ by _ directly in addition, and become a source of vitality consumed by the tribute. Volatile acid (in view), can = propionic acid. Moreover, Niu Wangqian The animal is called a fatty acid animal. The animal is called (four) fatty acid to obtain the energy source of the fat stomach = 3 protein 'in a single stomach animal such as pigs, the protein of the feed in the digestive acid is transported to the intestines' borrowed in the intestines It is decomposed into an amino acid by the action of the animal itself, and then the 'reverse substance' is decomposed into an amino acid by the microbial protein 2100108238 F TM 201136528 in the rumen, and then decomposed into ammonia, and then decomposed into ammonia. The microorganism uses the ammonia as a material to re-synthesize a microbial protein (bacterial protein, bombardment protein) which is essential for amino acid, that is, high nutritional value. The "microbial protein" thus formed is together with the stomach contents. After being transported to the fourth stomach, the process of being mainly decomposed into an amino acid in the small intestine and being absorbed is carried out in the same manner as the monogastric animal. Here, there is another path for digestion and absorption of proteins such as cattle, which is not in the rumen. A bypass protein that is digested by microorganisms and transported directly to the small intestine where it is digested. This is called a metabolic protein together with a microbial protein, but it is The intestine becomes an amino acid and is absorbed. Shi 0 4匕, 嚷乐~ stomach (rumen) is a so-called living fermentation device, supplying the necessary energy source, and another ~+. In terms of the type of fermentation by-products Methane or carbonic acid gas, nitrogen and small θ y are generated in % long > 1 oxygen, argon, hydrogen sulfide, etc., which are hardly used and are discharged into the atmosphere as a gas, thereby causing energy loss. Further, in the first stomach The nutrients in the feed are not enough to make the organic matter such as nitrogen in the feces, or minerals such as minerals, which are not used and are discharged. In addition, it is generally considered that one-third of the nitrogen taken up by the feed becomes feces. One third is discharged into the urine, and the rest is transferred to the milk or accumulated in the body. k When the healthy livestock with increased pore volume is raised, it is important to improve the growth or activation of the rumen The increase in the production of high energy-energy propionic acid or the increase in the digestibility of the protein protein from the protein source of nutritional value, the improvement of the digestibility of the feed, the reduction of energy loss, and the improvement of feed utilization efficiency require the development of such an effective anti-animal feed. Use efficiency Improve 1,001,082,388,201,136,528
[專利文獻1]曰 [專利文獻2]曰 [專利文獻3]曰 [專利文獻4]曰 [專利文獻5]曰 [專利文獻6]曰 本專利特開平2-255047號公報 本專利特開平6-237701號公報 本專利特開昭62-232434號公報 本專利特開2008-187929號公報 本專利特開平5-146260號公報 本專利特開平11-332555號公報[Patent Document 1] 专利 [Patent Document 2] 曰 [Patent Document 3] 曰 [Patent Document 4] 曰 [Patent Document 5] 专利 [Patent Document 6] 曰 Patent Patent Publication No. 2-255047 Japanese Patent Publication No. Hei. No. Hei. No. Hei. No. Hei. No. Hei. No. Hei. No. Hei. No. Hei. No. Hei.
[非專利文獻1]佐藤末太郎，，河—其之13—乳牛之生理特 異性之2與單胃動物之比較”，[online]，LIAJNEWS Νο·65(2000·9·25) ’ [平成22年2月16日檢索]，網際網路 <URL . http://liaj.lin.gr.jp/japanese/liajnews/65/6560.html> 【發明内容】 (發明所欲解決之問題） 有鐘於此種狀況’本發明似提供反㈣物之飼料利用效 率改善劑為目的。尤其是本發明係以提供反g動物之含固形 =飼料時之飼料彻效率改善劑為目的。再者，本發明係以 提供—種飼_収率改㈣之使財法為目的，其特徵在 於將飼料利用效率改善劑使用作為乳量改善劑。又，本發明 於提供—種反离動物之飼育方法，其特徵為將飼料 (解決問題Γ手t接或添加至飼料中而給予至㈣動物。 本案發明者等人經心研究，結果藉㈣物實驗等確認 100108238 201136528 到，藉由使用枯草桿菌（Bacillus subtilis)之生菌體作為有效 成分，則改善反羁動物之特異消化吸收機構或能量代謝機構 整體，亦即，改善反海動物之瘤胃醱酵並促進瘤胃内微生物 之增加及/或活性化，且提升瘤胃内之丙酸生成、微生物體 蛋白質之合成，再者，體内之有用菌增加而消化吸收效率提 升，使飼料之消化率、尤其是含有固形物之飼料的消化率提 升，減少能量損失，遂完成本案發明之反羁動物之飼料利用 效率改善劑。 本發明係以以下（1)至（10)記載之反g動物之飼料利用效 率改善劑為要旨。 ⑴-種反_物之飼料利用效率改善劑，其特徵為含有 枯草桿菌（Bacillus subtilis)之生菌體作為有效成分。 ⑺如⑴之飼料效率改善劑，其巾，飼料利用效率改 善係伴隨著飼料消化率提升。 (3)如⑴或(2)之飼料利用效率改善劑，其中，飼料利用效 率改善係伴隨著瘤胃醱酵改善。 (4)如⑴至(3)中任-項之铜料利用效率改善劑，其中，飼 料利用效率改善係伴隨著丙酸生成提升。 (5)如（1)至(4)中任-項之飼料利用效率改善劑^ 料利用效率改善係伴隨著微生物體蛋白質合成提升。 ⑹如⑴至(5)中任—項之飼料利用效率改善劑，其中， 料利用效率改㈣伴隨著體内有_之減少及有用菌之. 100108238 201136528 加。 ⑺如⑹之飼料利用效率改善劑，其中，有用g為乳酸桿 菌及雙叉乳酸桿菌（bifidus)。 于 (8) 如（1)至（7)中任一項之飼料利用效率改善劑，其中，3 料利用效率改善係伴隨著乳量改善。 飼 (9) 如（1)至（8)中任一項之飼料利用效率改善劑其中， 草桿菌為枯草桿菌C-3102(FERM BP-1096；)。 (1〇)如⑴至（9)中任一項之飼料利用效率改善劑， 反芻動物為牛、綿羊、山羊之任一種。 再者’本發明之要旨為以下(11}記載之使用方 載之反舞動物之飼育方法。 ）°己 (11) 一_料利収率改善狀使財法 ⑴至（10)中任一項之飼料利用效率改善劑作為;支為使用 (12) -種反_物之飼 1里改善劑。 任一項之铜料利用效率改善劑直接、=，將⑴至⑽中 至反离動物。 次添加至铜料中而給予 (發明效果） 本發明可提供—種反离動物之飼料 量改善劑’其特徵為，不需使用抗生物=善劑或乳 等’改善反咖之瘤胃—胃内 100108238 〜日刀口及 201136528 /或活性化，且提升瘤胃内之丙酸 合成，進而減少體内有害菌且增 / ^勿體蛋白質之 率，提升飼料之消化率、尤盆是含有固用菌而提升消化吸收致 減低能量損失。 固形物之飼料的消化率， 藉由將本發明之改善劑直接或添加於飼料中心 畜之反海動物，則發揮可铜育出飼料利用效 ^ 之健康家畜的效果。 里！改善 藉由將本發明之改善劑直接或添加至飼料中而 善反舞動物之瘤胃醱酵並提升^ 、° ，則改 合成，故使瘤胃液中之毒性強之氨濃度減少，而體内有= 減少且有用菌增加’故提升消化吸收效率，發揮可飼育^ 康且乳質改善之家畜的效果。 藉由將本發明之改善劑直接或添加至飼料中而給予則可 提升飼料利用效率、亦即提升铜料之消化率與減低能量損 失，故可減少所給予的飼料量，提升生產性而亦有利於經 濟’並可改善家畜之排出物量與f，而發揮有助於環境改善 的效果。 【實施方式】 本發明中，作為必須之有效成分而含有的生菌體為枯草桿 菌（Bacillus subtilis) ’ 其菌學性質記載於 Bergey’s Manual of[Non-Patent Document 1] Sato Taro, River - 13 of the physiological specificity of dairy cows compared with monogastric animals", [online], LIAJNEWS Νο·65 (2000·9·25) '[Heisei 22 Search on February 16th], Internet <URL. http://liaj.lin.gr.jp/japanese/liajnews/65/6560.html> [Summary of the Invention] (Problems to be Solved by the Invention) In this case, the present invention is intended to provide a feed utilization efficiency improving agent for the anti-(four) substance. In particular, the present invention is directed to providing a feed efficiency improving agent for the anti-g animal solid content=feed. The present invention is directed to providing a feed-for-yield (four)-based financial method, characterized in that a feed utilization efficiency improving agent is used as a milk amount improving agent. Further, the present invention provides a method for breeding a reversed animal. It is characterized in that the feed (the problem is solved or added to the feed and is given to the animal). The inventors of the present invention and others have studied the results, and the results are confirmed by the (four) experiment, etc., by using the Bacillus subtilis ( Bacillus subtilis) as an active ingredient, Good ruminant animal specific digestion and absorption mechanism or energy metabolism mechanism, that is, improve the rumen fermentation of anti-sea animals and promote the increase and / or activation of microorganisms in the rumen, and increase the production of propionic acid in the rumen, microorganisms The synthesis of protein, in addition, the increase of useful bacteria in the body and the improvement of digestion and absorption efficiency, so that the digestibility of feed, especially the digestibility of feed containing solids, is improved, and the energy loss is reduced, and the ruminant of the invention of the present invention is completed. The feed utilization efficiency improving agent according to the following (1) to (10) is characterized in that the feed utilization efficiency improving agent of the anti-g animal is characterized in that it contains a herbicide. (7) The feed efficiency improvement agent of (1), the improvement of feed utilization efficiency is accompanied by an increase in feed digestibility. (3) Feed utilization efficiency as (1) or (2) The improving agent, wherein the improvement of the feed utilization efficiency is accompanied by the improvement of the rumen fermentation. (4) The copper utilization efficiency improving agent according to any one of (1) to (3), wherein The improvement of the utilization efficiency of the material is accompanied by an increase in the production of propionic acid. (5) The utilization efficiency improvement of the feed utilization efficiency improver according to any one of (1) to (4) is accompanied by an increase in the synthesis of the microorganism protein. (6) (1) To the feed utilization efficiency improver of (5), the feed utilization efficiency is changed (4) with the decrease of the body and the useful bacteria. 100108238 201136528 Add. (7) The feed utilization efficiency improver of (6), wherein Useful g is Lactobacillus and Bifidus. (8) The feed utilization efficiency improving agent according to any one of (1) to (7), wherein the improvement in the utilization efficiency of the three materials is accompanied by an improvement in the amount of milk. (9) The feed utilization efficiency improving agent according to any one of (1) to (8) wherein the bacterium is Bacillus subtilis C-3102 (FERM BP-1096;). (1) The feed utilization efficiency improving agent according to any one of (1) to (9), wherein the ruminant is any one of a cow, a sheep, and a goat. Furthermore, the gist of the present invention is the breeding method of the anti-dance animal which is described in the following (11}.) ° (11) A material yield improvement is improved by any of the methods (1) to (10) The feed utilization efficiency improver of the item is used as the support agent for the use of (12)-the anti-substance feed. Any one of the copper materials utilizes the efficiency improving agent directly, =, from (1) to (10) to the reverse animal. Sub-addition to copper material for administration (effect of the invention) The present invention can provide a feed amount improving agent for a reverse animal, which is characterized in that it is not necessary to use an antibiotic, a good agent or a milk, etc. 100108238 ~ Nippon Knife and 201136528 / or activated, and promote the synthesis of propionic acid in the rumen, thereby reducing harmful bacteria in the body and increasing the rate of protein, improve the digestibility of feed, the pot contains solid bacteria Improve digestion and absorption to reduce energy loss. The digestibility of the feed of the solid matter is achieved by directly or adding the improving agent of the present invention to the anti-sea animals of the feed center, thereby exerting the effect of the healthy livestock capable of utilizing the feed efficiency of the copper. in! Improve the rumen fermentation of the animal by directly or adding the improving agent of the present invention to the feed, and improve the synthesis of ^, °, then the synthesis, so that the toxic concentration of ammonia in the rumen fluid is reduced, while the body There is = reduction and increase in useful bacteria', so the digestion and absorption efficiency is improved, and the effect of livestock that can be fed and improved in milk quality is exerted. By directly or adding the improving agent of the present invention to the feed, the feed utilization efficiency can be improved, that is, the digestibility of the copper material can be improved and the energy loss can be reduced, thereby reducing the amount of the feed and improving the productivity. Conducive to the economy' and can improve the amount of livestock emissions and f, and play the role of contributing to environmental improvement. [Embodiment] In the present invention, the bacterial cell contained as an essential active ingredient is Bacillus subtilis. The bacteriological properties are described in Bergey’s Manual of
Bacteriology Vol.ll(1986)。本發明中，可使用公知之任一種 枯草桿菌，較佳可舉例如枯草桿菌C-3102等。 100108238 12 201136528 屬；_之括草桿菌（Bacillus subtilis)C-3102 菌株，伟 於1985年12月25日，以寄存編號：微工研菌寄第μ料 5虎原寄存於日本國茨城縣筑波市東1丁目1番地1筑波中心 中央第6(郵遞區號3〇5_5866)[寄存時：日本國茨城縣筑波郡 合田和丁東1 丁目1番地3(郵遞區號305)]之獨立行政法人 產業技術總合研究所糊生物寄存K寄存時：通商產業 省工業技術院微生物工業技術研究所]，並於 1986年6月 28日由同機構依寄存編號FERM Bp i()96(移管時：微工研 條寄第10%號）移管為國際寄存，又，枯草桿菌 C 3102(FERMBlM()96)^gi^性質已公開（專利文獻3)。 於土。養上述枯草桿菌時，可使用於微生物培養時通常使用 之含碳源、氮源、無機物等之液體培養基或HI體培養基作為 培養基。作為&源，若為枯草桿时利用的碳源即可可舉 例如葡萄糖、果糖、蔗糖、殿粉、糖蜜等，又，作為氮源， 可舉例如胜肽路蛋自水解物、肖萃取物U找等。再者， 視需要亦可添㈣酸、鉀、鎂，、鈉、鐵及猛等之鹽類、 _命類、胺類、界面活性解。作為培養條件，較佳係依 好樣條件下進仃’培養裝置較佳係❹例如由缸式酿酵槽 所進行的通氣攪拌液體培養、棚式固體培養、自動製麵培養 裝置等。培養溫度較隹為20〜耽、特佳為30〜45ΐ，典養 時間為12小時〜7天’培養阳為5〜9、特佳為6〜8。 上述所培養的培養物，可將培養物本身、其濃縮物或由其 100108238 13 201136528 所分離出之菌體直接作為有效成分。此 守’有效成分中可合 有例如枯草桿菌C-31〇2(FERM BP-l〇96w 株之;Ψ喆舻、會p口 芽胞及/或營養細胞1〇〜1〇!2個/g、較佳 ” 〜10u 個/e。 而且，在調製本發明之反㈣物之飼料利用效率、乳量改 善劑時，較佳係將上述有效成分直接、 千孔里汉 , ^ A1 或视需要添加賦形 劑，糟由乾燥或製劑化之方法等而獲得，作亦可將、乂 ^ 成分直接作為改善劑。該賦形劑並無特 " 丨艮疋，可使用例如 碳酸鈣、脫脂米糖、玉米粗粉、玉米粉、 i 4 麥麩、脫脂奶粉等。 本發明之反_物之侧利収率投㈣心 無特別限定’可直接給予反嶋，較佳可作成粉末、：立 是微粉末等，並添加、混合至公知飼料中 ’ . 使用。在添加混 合至公知飼料時之本發明之改善劑的添加量，係依家畜種 類、曰齡等而異’較佳係依例如添加至例料後之枯 «1〇2(FERMBP_脳）之生菌數丨1〇3〜1〇9㈣、2 104〜108個/g之方式進行添加、混合。Bacteriology Vol.ll (1986). In the present invention, any of the known Bacillus subtilis can be used, and preferably, for example, Bacillus subtilis C-3102 or the like can be used. 100108238 12 201136528 genus; _ Bacillus subtilis C-3102 strain, Wei on December 25, 1985, with the registration number: Micro-Working bacteria sent the first material 5 Hu Yuan was deposited in Tsukuba, Ibaraki Prefecture, Japan City, 1st, 1st, 1st, 1st, 1st, Tsukuba, the center of the city, the 6th (postal area code 3〇5_5866) [At the time of the deposit: the industrial and technical system of the independent administrative corporations of the Higashi-ku, Ibaraki, Izumi, Izumi, Izumi, Izumi, Izumi, Izumi, Izumi, Izumi When the research institute pastes the bio-storage K deposit: Institute of Microbial Industrial Technology, Institute of Industrial Technology, Ministry of Industry and Commerce, and on June 28, 1986, the same institution is registered under the number FERM Bp i () 96 (transfer tube: micro-work Article 10%) The transfer tube is internationally registered, and the B. subtilis C 3102 (FERMB1M() 96) ^gi^ property has been disclosed (Patent Document 3). In the soil. In the case of the above Bacillus subtilis, a liquid medium or a HI medium containing a carbon source, a nitrogen source, an inorganic substance or the like which is usually used for microbial culture can be used as the medium. As the source of the & source, the carbon source used in the case of the dry grass can be, for example, glucose, fructose, sucrose, sucrose, molasses, etc., and examples of the nitrogen source include peptide peptide autolyzate and extract. U looking for. Furthermore, it is also possible to add (iv) acid, potassium, magnesium, sodium, iron, and sulphate salts, scorpions, amines, and interfacial active solutions. As the culture conditions, it is preferred to carry out the cultivating apparatus under the conditions of the cultivating apparatus, for example, aeration agitation liquid culture, a shed solid culture, an automatic noodle culture apparatus, and the like by a tank type fermentation tank. The culture temperature is 20 to 耽, particularly preferably 30 to 45 ΐ, and the cultivation time is 12 hours to 7 days. The culture yang is 5 to 9, and the optimum is 6 to 8. The culture to be cultured above can directly use the culture itself, its concentrate or the cells isolated by its 100108238 13 201136528 as an active ingredient. This defensive 'active ingredient can be combined with, for example, Bacillus subtilis C-31〇2 (FERM BP-l〇96w strain; Ψ喆舻, will p-spore and/or vegetative cells 1〇~1〇! 2/g Preferably, it is ~10u/e. Moreover, when preparing the feed utilization efficiency and the milk amount improving agent of the anti-(four) substance of the present invention, it is preferred to directly use the above-mentioned active ingredient, a thousand pores, ^ A1 or as needed The excipient is added, and the residue is obtained by a method of drying or formulation, etc., and the component can be directly used as an improving agent. The excipient is not particularly useful, and for example, calcium carbonate or degreasing can be used. Rice candy, corn meal, corn flour, i 4 wheat bran, skim milk powder, etc. The side yield of the anti-substance of the present invention (four) heart is not particularly limited 'can be directly given ruminant, preferably can be made into a powder,: It is a fine powder or the like, and is added and mixed into a known feed. 'Use. The addition amount of the improvement agent of the present invention when added to a known feed varies depending on the type of livestock, age, etc.' For example, the number of bacteria of the dried «1〇2 (FERMBP_脳) added to the sample 丨1〇3~1〇9(4), 2 104 108 / g of the manner added and mixed.
另外，本發明之反S動物之铜料利用效率改善劑的有 取量係視反㈣物之種類、日齡等而異，較佳為依例如 桿菌《啊舰則㈣96)之生菌數成為lxl06個/κ 以上、特佳5x106〜1X108個化•日之方式進行攝取。再者曰 投予時期亦視反㈣物之種類、日齡等而異，但 投予。例如，在牛的情況，對使用含固形物飼料之離乳續 或離乳後之育成牛進行投予則特別有效。仔牛、尤其是哺A 100108238 14 201136528 中之(尚未吃濃厚顺、粗飼料、其他固形飼料等之固形物） 仔牛中’牛胃尚未分化，雖存在(將形成)瘤胃的部位但尚不 發達。而'^ ’僅攝取牛乳或液狀物的情況，瘤胃人口關閉而 液體在物體無法進人瘤胃中的狀態下流人，成為與人類或緒 同樣之單胃動物相同的狀態，若開始食用固形物，則對其有 所反應而瘤胃人口開放。如此，藉由開始食賴形物，必須 強化反舞之消化(分解)力，而第—胃（瘤胃）逐漸發達，故在 此時期給予本發明之改善劑以改善、調整瘤胃醱酵，促進瘤 月内之原生動物等微生物的增加活性化，而將對其後成長 發揮極大效果。 作為本發明對象的反离動物的麵並無特別限定，較佳可 舉例如牛、料、山羊等乳成為商品的反努動物JL飼養作為 豕畜者。又’本發明之飼料彻效率改善劑亦可使用作為乳 量改善劑’對象並不限定於乳牛等。 以下藉實施例及試驗例說明本發明之細節及效果，但本發 明並不受此等實施例等任何限制。 [實施例1] (生菌末及飼料利用效率改善劑之製造） 於自來水10公升中溶解大豆腺2〇〇g、磷酸二鉀1〇g及糖 蜜200g❿調製培養|，再添加1N氮氧化納水溶液使培養 基之pH調整為7.5後，裝入虹式酸酵槽中，依12Γ(：、15 分鐘進行殺菌。接著，冷卻至坑，取事先培養之枯草桿 100108238 15 201136528 菌C-3102(微工研條寄第1096號)之培養液】，依37°c 進行通氣振盪培養40小時。接著將所得培養液離心並收集 菌體，使其乾燥後，混合碳酸鉀180g而製造反籍動物之飼 料利用效率改善劑（以下稱為「改善劑1」）2〇〇g。所得改善 劑1中之枯草桿菌C-3102(FERMBP-l〇96)的生菌數為 lxl01Q 個/g。 ⑴離體(In vitro)試驗；試驗例1 使用貫施例1所得之改善劑1，依以下所示試驗條件進行 離體試驗。 (培養液）：由泌乳牛在早晨給飼前使用導管採取瘤胃液， 將過遽後之瘤胃液與緩衝液依1:2比例稀釋而調製培養液。 (基質）：將濃厚飼料(森永乳業製「乳牛用調配飼料」）與 乾草(蘇丹草)依1 : 1混合而調製飼料，並作為基質。 (試驗試料)：準備於基質中分別添加了改善劑上 60PPm、9〇Ppm的血清瓶(6〇1111容量）。該添加了 3〇ppm、 6〇PPm、90PPm之試料係分別於基質（混合飼料）中添加了生 菌3χ105個/g、6χΐ05個/g、9x105個/g者。將僅有基質者(未 添加改善劑1)作為對照。 (试驗方法）·於各試料瓶中加入上述培養液’進行氣 換，依39°C進行厭氣培養6小時。6小時後測定pH 氣體、氫氣等。 成 (結果） 10010S238 201136528 PH值、甲烷氣體發生量係對照與各試料中均無變化。 關於氫氣發生量則有變化，見到氫氣發生量與改善劑工 添加畺成反比地減少之傾向。結果示於表1。 [表1] 濃度(ppm) 氫氣發生量（°/〇) 0 100.0 30 ---- 80.9 60 -~~~--- 88.2 90 72.3 ~ ----' ------ (結果檢討) pH值未變化係顯示藉由改善劑1之添加，而不致發生過 度醱酵，揮發性脂肪酸(VFA)的產生平衡未失衡，不需擔心 瘤胃酸中毒發生。而且，在反海動物藉由瘤胃醱酵，將碳水 化合物分解而作成揮發性脂肪酸（VFA)時、生成曱酸、乙 酸、丁酸時’雖發生氫氣’但由在生成丙酸時係、消耗氣的技 術常識而言’此試驗例i中有關氫氣的結果(藉由添加改善 而氫氣發生畺減少）顯#，揮射生脂肪酸中丙酸生成的比 例增加。 揮發性脂猶制於錢房巾形成乳成分的能量源（參照 非專利文獻1)，該丙酸已知係於揮發性脂肪酸中能量效率 最高的酸，而且為在瘤胃吸收佳的酸。 因此，此試驗例1之結果可謂顯示出本發明之改善劑的瘤 100108238 17 201136528 胃_改善或其所造成之飼料則鱗改善及乳量改善。 接著飼養實際的牛，確認其效果。 (H)活體内（In vivo)試驗；試驗例2~7 /吏用由實關1所得之轉劑1，依以下所我驗條件進 仃活體内試驗(動物實驗）。 =牛广平均趙重約160Kg，日齡約4個月的育成牛4 員u毛和牛之雄性：2頭’荷士登之雄性：2頭） (基本=將濃厚飼料(森永乳業製「乳牛用調配飼料」) 二驗: 料：乾草=1 : A混合的飼料。 (试驗期飼料）：相對於基本飼 的飼料(添加至飼料的生菌數成為…^ ^其中連續給予2周的試驗 給予2周之基本詞料(對照期2)。4(4驗期），之後再連續 ⑴試驗例2<關於飼料消化率之提升> 依上述試驗條件進行試驗，於對 2之各期後半採取各頭牛的糞“、、月、拭驗期、對照期 其重量(糞固形物重量）。（全量）’使糞完全乾燥，測定 另外’亦將基本飼料、試驗期飼。 (飼料固形物重量）。（全糞採取、疋王乾燥，測定其重量 設為:飼料消化率(%)，二）， 重量）」Χ]〇0 ,分别算出對昭期固形物重量)/(旬科固形物 _38 ( 2)與·_平均消化 201136528 率。該值以平均值±標準偏差所表示（以下試驗例中亦相 同）。此消化率通常稱為DM(DRY MATTER)消化率。結果示 於表2及圖1。 [表2] 對照期 試驗期 飼料消化率(％) 58.6±2.1 62_2 士 0.9 (結果檢討) 飼料消化率（％)係對照期1、2之平均為58.6%，試驗期之 平均為62.3%，於飼料消化率為使用了改善劑1的試驗期 中，可見到有意義(p< 0.05)提升。因此，藉由添加了本發明 之飼料利用效率改善劑的飼料，確認到飼料消化率提升。 飼料消化率之提升表示飼料的有效利用，顯示乳房中用於 形成乳成分的能量源增加，故本結果可謂顯示了藉由使用本 發明之飼料利用效率改善劑而改善了飼料利用效率及乳量。 (2)試驗例3<關於瘤胃液中之丙酸莫耳比率增加〉 依上述試驗條件進行試驗，於對照期1、試驗期、對照期 2之各期最終日採取各自的瘤胃液，分析液中之各揮發性脂 肪酸(VFA)，分別計算總揮發性脂肪酸量(mM)、相對於總揮 發性脂肪酸之乙酸、丙酸、丁酸的各莫耳比率(mol%)並算出 各平均值。結果示於表3，將其中有關丙酸莫耳比率的結果 示於圖2。 100108238 19 201136528 [表3] 對照期 試驗期 總揮發性脂肪酸量 65.1±15.4 67.8 士 13.5 (mM) 乙酸莫耳比率(mol%) 67·0士3.2 68.0 士 2.8 丙酸莫耳比率(mol%) 17.0±1.3 18.3±1.3 丁酸莫耳比率(mol%) 12.4±2.3 9.7±2.4 (結果檢討) 關於瘤胃液中之總揮發性脂肪酸量、乙酸莫耳比率，對照 期與試驗期的變化率小，並未確認到有意義的差。 另一方面，瘤胃液中之丙酸莫耳比率，係對照期1、2之 平均為17.0%、試驗期之平均為18.3%，於使用了改善劑1 之試驗期中確認到有意義（p<〇.001)的增加。又，丁酸莫耳 比率係對照期1、2之平均為12.4%、試驗期之平均為9.7%， 於使用了改善劑1之試驗期中確認到有意義（p< 0.001)減 〇 此結果可謂是與離體試驗(試驗例1)結果整合者。 藉由使用本發明的飼料利用效率改善劑，則改善、調整瘤 胃醱酵，不生成過度之揮發性脂肪酸量，使其脂肪酸組成變 化為良好組成，此結果驚人。 一般技術常識已知，揮發性脂肪酸(VFA)係由瘤胃壁直接 被吸收，於乳牛等中成為最終消化產物之乳成分等的原料， 100108238 20 201136528 此外，並成為貢獻至體組織所消耗之能量的6成以上、乳房 中組成乳成分時所使用之燃料之約一半的活力源（參照非專 利文獻1)，而揮發性脂肪酸(VFA)中，能量效率最高者為丙 酸，亦為由瘤胃吸收佳的酸，故本結果（丙酸莫耳比率增加) 可謂顯不了藉由使用本發明之飼料利用效率改善劑，將改盖 飼料利用效率’其結果改善乳量。 (3)試驗例4<關於瘤胃液中之氨態氮濃度的減少（微生物體 蛋白質合成之提升)> 依上述試驗條件進行試驗，於對照期卜試驗期、對照期 2之各期的最終日分職取瘤胃液，測定液中之氨態氣濃产 (m_)，Μ賴期（卜2)與試驗敵各卿均值。結果二 於表4及圖3。 。不 [表4] ΝΗ3_Ν濃度(mg/叫 (結果檢討） 對照期 ---~--- 試驗期 5.7 土 2.2 2-5±1.2 -—~~~~~~_ 瘤月液中之氨態氮濃度(mg/dl)，係對照期卜a之平均為 驗期之平均為2 5 ’㈣了改善劑丨之試驗期中確認 到氰恶気濃度有意義(p<0.001)的減少。 —一般技術常識已知，於牛等反_物巾，飼料中的蛋白源 1由瘤胃内微生物的蛋白分解酵素之作用而分解為胺基 酸’再分解城’進—以錄騎料以合成高營養價 100108238 21 201136528 離物體蛋白質（參照非專利文獻1V因此，本結果(氨 稽 <減））係顯示出由瘤胃内微生物所進行之高營養 心物體蛋白質的合成提升，並顯示出由飼料攝取之 、致率的提升(能量損失之減少）。微生物體蛋白質於 内的解吸收，成為乳成分之原料等。再者，已知若體 明之11?多’則氨之強毒性亦導致中毒，故藉由使用本發 提升。H ’乳有意義地減少，故可謂健康提升且吸收力亦 劑而改善^係精由使用本發明之飼料顧效率改善 ⑷試驗t=T其結果改善乳量。 j5<關於瘤胃内微生物 依上述試驗條件進行試驗，_二/或活性化〉 試驗期、斜照期2之各期的最终曰::牛3頭’於對咖 給飼料“小時、給飼料後5小時^]於^料前0小時、 原生動物數量，分別整合對_们碌瘤讀，測定液中之 值與試，之各日請的所相所有測定 表5及圖4。 异出平均值。結果示於 [表习Further, the amount of the copper material utilization efficiency improving agent of the anti-S animal of the present invention varies depending on the type of the anti-(four) substance, the age, and the like, and preferably the number of bacteria according to, for example, the bacillus "Ark (4) 96) becomes Lxl06 / κ or more, especially good 5x106 ~ 1X108. In addition, the period of investment will also vary depending on the type of anti-(four) objects, age, etc., but will be given. For example, in the case of cattle, it is particularly effective to administer a bred cow after the use of a solid-containing feed or after weaning. Cattle, especially in A 100108238 14 201136528 (has not eaten solids such as thick, roughage, other solid feeds, etc.) In the calves, the beef stomach has not yet differentiated, although it is (will form) the rumen, but it is not yet developed. When '^' only takes milk or liquid, the rumen population is closed and the liquid flows in a state where the object cannot enter the rumen, and becomes the same state as the human or the same monogastric animal. If the solid substance is started to be eaten, , it responds to it and the rumen population is open. In this way, by starting to eat the food, it is necessary to strengthen the digestion (decomposition) force of the anti-dance, and the first stomach (rumen) is gradually developed, so the improvement agent of the present invention is given at this time to improve and adjust the rumen fermentation and promote The increase in the number of microorganisms such as protozoa in the tumor month will greatly increase the growth of the microorganisms. The surface of the animal to be degraded as the object of the present invention is not particularly limited, and it is preferable to use, for example, a cow, a feed, a goat, or the like as a commercial product. Further, the feed efficiency improving agent of the present invention can be used as a milk amount improving agent, and is not limited to cows and the like. The details and effects of the present invention are illustrated by the following examples and test examples, but the present invention is not limited by the examples and the like. [Example 1] (Production of a bactericidal agent and a feed utilization efficiency improving agent) Dissolve 2 g of soybean gland, 1 g of dipotassium phosphate, and 200 g of molasses in 10 liters of tap water to prepare culture| After the pH of the medium was adjusted to 7.5 by an aqueous solution, it was placed in a rainbow acid fermentation tank and sterilized by 12 Γ (:, 15 minutes. Then, it was cooled to a pit, and the previously cultivated dry grass rod 100108238 15 201136528 bacterium C-3102 (micro The culture medium of the No. 1096) was cultured at 37 ° C for 40 hours, and then the obtained culture solution was centrifuged, and the cells were collected, dried, and then mixed with potassium carbonate (180 g) to produce a reverse animal. The feed utilization efficiency improving agent (hereinafter referred to as "improving agent 1") was 2 g. The number of bacteria of Bacillus subtilis C-3102 (FERMBP-l〇96) in the obtained improving agent 1 was lxl01Q/g. In vitro test; Test Example 1 Using the improving agent 1 obtained in Example 1, the ex vivo test was carried out according to the test conditions shown below. (Culture): The rumen fluid was taken from the lactating cow before feeding in the morning. , the rumen fluid and buffer after the sputum are diluted 1:2 The culture medium is prepared. (Matrix): The feed is prepared by mixing the thick feed ("Yangyong Dairy" "mixed feed for dairy cows") with hay (Sudan grass) according to 1:1. (Test sample): Prepared for the substrate A serum bottle (6〇1111 capacity) of 60 ppm and 9 〇 Ppm on the improver was added, and the sample added with 3 〇 ppm, 6 〇 PPm, and 90 ppm was added to the substrate (mixed feed). /g, 6χΐ05/g, 9x105/g. Only the matrix (no additive 1 added) was used as a control. (Test method) · Add the above culture solution to each sample bottle for gas exchange. Anaerobic culture was carried out for 6 hours at 39 ° C. After 6 hours, pH gas, hydrogen gas, etc. were measured. (Result) 10010S238 201136528 PH value, methane gas generation amount control and no change in each sample. The change showed a tendency to reduce the amount of hydrogen generated in an inversely proportional manner to the increase in the addition of hydrazine. The results are shown in Table 1. [Table 1] Concentration (ppm) Hydrogen generation amount (°/〇) 0 100.0 30 ---- 80.9 60 -~~~--- 88.2 90 72.3 ~ ----' ------ (Result review) pH The unaltered line shows that the addition of the improving agent 1 does not cause excessive fermentation, and the balance of the production of volatile fatty acids (VFA) is not unbalanced, and there is no need to worry about rumen acidosis. Moreover, the anti-sea animals are fermented by the rumen. When the carbohydrate is decomposed to form a volatile fatty acid (VFA), when citric acid, acetic acid, or butyric acid is formed, 'hydrogen is generated, but the technical common sense is that when propionic acid is generated and gas is consumed, this test example i As a result of the hydrogen gas (reduced by the addition of hydrogen, the hydrogen generation is reduced), the proportion of propionic acid in the raw fatty acid is increased. The volatile fat is still produced as an energy source for forming a milk component of the money towel (refer to Non-Patent Document 1), which is known to be the most energy-efficient acid among volatile fatty acids, and is an acid which is well absorbed in the rumen. Therefore, the result of this Test Example 1 can be said to show the improvement agent of the present invention. 100108238 17 201136528 Stomach_improvement or the resulting scale improvement of the feed and improvement in milk amount. Then raise the actual cattle and confirm the effect. (H) In vivo test; Test Example 2~7/吏 Use the transfer agent 1 obtained from the actual test 1 and perform the in vivo test (animal test) according to the following conditions. = Niu Guang average Zhao weight about 160Kg, age of about 4 months of breeding cattle 4 members u hair and cattle male: 2 heads 'Hersden's male: 2 heads) (Basic = will be thick feed (Song Yong Dairy "dairy cow" Use the blended feed") Second test: Material: Hay = 1: A mixed feed. (Test feed): Relative to the basic feed (the number of bacteria added to the feed becomes ... ^ ^ which is continuously given for 2 weeks) The test was given 2 weeks of basic vocabulary (control period 2). 4 (4 test period), followed by continuous (1) test case 2 <improvement of feed digestibility> Test according to the above test conditions, in the period of 2 In the second half, the feces of each cow were taken, “, month, wipe test period, and weight of the control period (weight of fecal solids). (Full amount) 'The feces were completely dried, and the other 'also measured the basic feed and the test period. Solid weight) (All the manure is taken, the king is dried, the weight is set to: feed digestibility (%), two), the weight is "Χ] 〇 0, respectively, the weight of the solids of the show is calculated) / (Ten Section solids _38 (2) and · _ average digestion 201136528 rate. This value is expressed as mean ± standard deviation (to The same is true in the next test example. This digestibility is usually called DM (DRY MATTER) digestibility. The results are shown in Table 2 and Figure 1. [Table 2] Feed digestibility during the test period (%) 58.6 ± 2.1 62_2 0.9 (Result review) The feed digestibility (%) was 58.6% for the control period of 1 and 2, and the average for the test period was 62.3%. The feed digestibility was meaningful during the test period using the improver 1 (p< 0.05). Therefore, the feed digestibility is improved by the feed of the feed utilization efficiency improver of the present invention. The increase in feed digestibility indicates the effective use of the feed, indicating the energy used to form the milk component in the breast. Since the source is increased, the present result shows that the feed utilization efficiency and the amount of milk are improved by using the feed utilization efficiency improving agent of the present invention. (2) Test Example 3 <About the increase in the molar ratio of propionate in the rumen fluid> The above test conditions were tested, and the respective rumen fluids were taken on the final day of each of the control period 1, the test period and the control period 2, and the volatile fatty acids (VFA) in the liquid were analyzed to calculate the total volatile fatty acid amount (mM). The respective average values were calculated for each molar ratio (mol%) of acetic acid, propionic acid, and butyric acid of the total volatile fatty acid. The results are shown in Table 3, and the results relating to the molar ratio of propionic acid are shown in Fig. 2. 100108238 19 201136528 [Table 3] Total volatile fatty acid content during the test period of the control period 65.1±15.4 67.8 ± 13.5 (mM) Molar ratio of mol (mol%) 67·0 ± 3.2 68.0 ± 2.8 molar ratio of propionate (mol%) 17.0±1.3 18.3±1.3 molar ratio of butyric acid (mol%) 12.4±2.3 9.7±2.4 (Result review) About the total volatile fatty acid amount in the rumen fluid, the molar ratio of acetic acid, the change rate of the control period and the test period is small , did not confirm the meaningful difference. On the other hand, the molar ratio of propionate in the rumen fluid was 17.0% in the control period, and the average in the test period was 18.3%, which was confirmed to be meaningful during the test period in which the improving agent 1 was used (p<〇 .001) increase. Further, the molar ratio of butyric acid was 12.4% in the control period of 1, 2%, and the average in the test period was 9.7%, which was confirmed to be meaningful in the test period in which the improving agent 1 was used (p < 0.001). Integral with the results of the in vitro test (Test Example 1). By using the feed utilization efficiency improving agent of the present invention, the rumen fermentation is improved and adjusted, and the amount of volatile fatty acid is not generated to change the fatty acid composition into a good composition. This result is surprising. It is known in the art that volatile fatty acids (VFAs) are directly absorbed by the rumen wall and become the raw material of the final digestion product in dairy cows, etc., 100108238 20 201136528 In addition, they become energy consumed by the body tissues. 60% or more of the vitality of about half of the fuel used in the formation of the milk component in the breast (see Non-Patent Document 1), and among the volatile fatty acids (VFA), the highest energy efficiency is propionic acid, also by the rumen Since the good acid is absorbed, the result (increased molar ratio of propionate) is remarkably improved by using the feed utilization efficiency improving agent of the present invention to change the feed utilization efficiency. (3) Test Example 4 <Reduction of the concentration of ammonia nitrogen in the rumen fluid (improvement of microbial protein synthesis)> The test was carried out according to the above test conditions, and the final period of each of the control period and the control period 2 The rumen fluid was taken in the day, and the ammonia gas concentration (m_) in the liquid was measured, and the mean period (b) and the average value of the test enemy. The results are shown in Table 4 and Figure 3. . No [Table 4] ΝΗ3_Ν concentration (mg/called (result review) Control period---~--- Test period 5.7 Soil 2.2 2-5±1.2 -~~~~~~~_ The ammonia state in the tumor liquid The nitrogen concentration (mg/dl), which is the average of the control period, is the average of the test period is 25'. (4) The reduction of the cyanide concentration in the test period of the improver 有意义 is meaningful (p<0.001). Common sense is known, in cattle and other anti-clothes, the protein source 1 in the feed is decomposed into the amino acid 're-decomposing city' by the action of the proteolytic enzyme of the microorganism in the rumen to record the riding material to synthesize high nutrient price. 100108238 21 201136528 From the object protein (refer to Non-Patent Document 1V, the present result (ammonia < subtraction)) shows the synthesis of high-nutrient heart protein by microorganisms in the rumen, and shows that it is ingested by the feed. Increase in the rate (reduction in energy loss). The absorption of the microbial protein inside is the raw material of the milk component. Furthermore, it is known that if the body is more than 11%, the strong toxicity of ammonia also causes poisoning. By using this hair boost, H 'milk is meaningfully reduced, so it can be described as healthy Improvement and absorption of the agent to improve the efficiency of the feed by using the feed of the present invention (4) Test t = T, the result of which improves the amount of milk. j5 <About the rumen microorganisms according to the above test conditions, _ two / or activation 〉 The final period of each period of the test period and the oblique period 2:: 3 heads of cattle 'hours to the coffee, 5 hours after feeding, ^ 0 hours before the feed, and the number of protozoa, respectively. We read the tumor, and measured the value in the liquid and the test, and all the measurement tables 5 and 4 of each day were used. The average value was shown.
原生動物數之總合时猶㈣期為3.赠個/mj，試 原生動物數(χ 105個/mJ) (結果檢討） 100108238 22 201136528 驗期為4.3xl〇5個/ml’使用了改善劑！ <忒驗期的原生^ 數的平均值為較高值。 足動％ 瘤胃内微生物（細菌與原生動物）係與痼田 一屬月醱酵有關， 白質分解，同時以屬於蛋白源分解物之氨作為材料，將冬 成出高營養價值之微生物體蛋白質（菌體晰’蔓_含 貧白貝、蟲㈣处 質），再者，原生動物等具有作為微生物 冬白 ㈣一…一…… 體蛋白質而與 内 容物-起被運送至第4胃以後並於小腸所消化吸收、 制’故原生動物數於瘤胃液中之變動較大，但於至少此， 例5的結果中，若與上述試驗例2之飼料消化率 ％ 驗例4之微生物體蛋白質合成之提升整合來看，可或巧 使用本發明之飼料利用效率改善劑所造成 礙^了 之增加及/或活性化之促進。 門礅生物 (5)Μ_ 6<_有㈣之減少及有用菌之增加〉 Τ述試驗條件進行試驗，於對照期卜試驗期、對 糞中/I ^半採取各牛的新鮮糞，依光岡法變法培養、測定 叢，針對各個期算出平均值。結果示於表6及圖5。 又，表中之ND仫主- 币表不檢測界限(2.3〇CFU/g)，數值（平均值土 標準偏差)下的(分數)表示檢測率。 100108238 23 201136528 [表6] 菌種 對照期1 試驗期 對照期2 大腸菌群 6.75±0.97 6.29±0.64 6.04±0.86 乳酸桿菌 3.74±0.21 4.28 2.92 士 0.47 (2/4) (1/4) (2/4) 雙叉乳酸桿菌 ND 5.16±0.04 ND (2/4) 厭氣性菌總數 6.91 土 0.71 6_42 士 0.47 6.49±0.56 枯草菌 4.08±0.03 5.15±〇.〇5 4.13±0.14 (結果檢討) 糞中菌叢之各菌種的菌數變化，係⑴屬於有害菌之大腸菌 群的菌數中，相較於對照期1，試驗期及對照期2呈減少； (ii)屬於有用菌之乳酸桿菌、雙叉乳酸桿菌之菌數，相較於 對照期1，試驗期呈增加，於其後之對照期2呈減少。尤其 是試驗期之雙叉乳酸桿菌的增加特別顯著。 僅在試驗期、亦即僅在本發明之飼料利用效率改善劑的使 用期中，屬於有用菌之乳酸桿菌、雙叉乳酸桿菌增加的本結 果令人驚訝。尤其值得注意的是，雙叉乳酸桿菌係於對照期 卜對照期2均為檢測界限以下(ND)，僅於試驗期顯著增加。 習知以來，已知藉由對飼養動物投予枯草桿菌，可抑制飼 養動物體内的病原性細菌之增殖（參照專利文獻4〜6)，但並 未得知有用菌之增加。又，乳酸桿菌、雙叉乳酸桿菌可良好 100108238 24 201136528 保持腸内㈣、提升吸收力、有助健康維持的事實已為眾所 皆知，但將此等菌由外部使用作為铜料添加物則有較為麻煩 專诸多問題(例如參照日本專利特開2__45號公報）。因 此，不由外部進行有用8之好，而㈣本發明之改善劑， 藉此可增加體时用㈣效料人，因此，使㈣提升且吸 收力亦提升。因此，本結杲(有㈣之減少及有㈣之增加） 可謂顯不了藉由使用本發明之飼料彻效率改善劑而改善 飼料利用效率，其結果亦改善乳量。 σ (6)試驗例7<關於曱烷氣體發生量> 依上述試驗條件進行試驗，於對照期i、試驗期、對照期 2之各期最終日，分職罩式呼吸試驗裝置測定甲燒氣辭 生量_MI/day) ’分別算出對照期（i、2)與試驗期的平^ 值，結果示於表7。 [表7] —---—η 對照期 曱烷氣體發生量(L/DMI/day) 33.4±3.3The total number of protozoa is (4) is 3. Give a /mj, try the number of native animals (χ 105 / mJ) (Result review) 100108238 22 201136528 The test period is 4.3xl 〇 5 / ml 'Used improvement Agent! <The average value of the original number of the test period is a higher value. Foot movement % The rumen microbes (bacteria and protozoa) are related to the genus of the genus Putian, and the white matter is decomposed. At the same time, the ammonia which is a protein source decomposition product is used as a material to produce a high-nutrient microbial protein in winter. The bacteria are clear, the vines contain the poor white shellfish and the worms (four), and the protozoa, etc., have the microbes as winter microbes (four) one...one body protein and the contents are transported to the fourth stomach and In the small intestine, the number of protozoa in the small intestine is greatly changed in the rumen fluid, but at least this, in the results of the example 5, if the feed digestibility of the above test example 2 is the microbial protein of the test example 4 In view of the improvement of the synthesis, it is possible to use the feed utilization efficiency improving agent of the present invention to cause an increase in the increase and/or promotion of activation. Threshold organisms (5) Μ _ 6 < _ have (four) reduction and increase of useful bacteria > Test conditions to test the test, in the control period, in the feces / I ^ half take the fresh feces of each cattle, Yi Guanggang The method was used to train and measure the plexus, and the average value was calculated for each period. The results are shown in Table 6 and Figure 5. Further, the ND 仫 main currency table in the table does not detect the limit (2.3 〇 CFU / g), and the value (score) under the numerical value (average standard deviation) indicates the detection rate. 100108238 23 201136528 [Table 6] Strain control period 1 Test period Control period 2 Coliform group 6.75±0.97 6.29±0.64 6.04±0.86 Lactobacillus 3.74±0.21 4.28 2.92 ± 0.47 (2/4) (1/4) (2/ 4) Lactobacillus bifidum ND 5.16±0.04 ND (2/4) Total anaerobic bacteria 6.91 Soil 0.71 6_42 ± 0.47 6.49±0.56 Subtilis 4.08±0.03 5.15±〇.〇5 4.13±0.14 (Result review) Feces The number of bacteria in each strain of the flora is (1) the number of bacteria belonging to the coliform group of the harmful bacteria, compared with the control period 1, the test period and the control period 2 are decreased; (ii) the lactobacillus belonging to the useful bacteria, The number of bacteria of Bifidobacterium bifidum increased compared with the control period 1, and decreased during the test period. In particular, the increase in lactobacilli in the test period was particularly remarkable. This result of an increase in the number of Lactobacillus and Bifidobacterium breve belonging to the useful bacteria was surprising only during the test period, that is, only during the use period of the feed utilization efficiency improving agent of the present invention. It is particularly noteworthy that the Lactobacillus bifidum was in the control phase 2 of the control period and was below the detection limit (ND), which was only significantly increased during the test period. Conventionally, it has been known that the growth of pathogenic bacteria in a feeding animal can be suppressed by administering Bacillus subtilis to a rearing animal (see Patent Documents 4 to 6), but an increase in useful bacteria is not known. Moreover, Lactobacillus and Lactobacillus bifidum can be well 100108238 24 201136528. The fact that the intestines (4) are maintained, the absorption is improved, and the health is maintained is well known, but these bacteria are used externally as copper additives. There are many problems that are more troublesome (for example, refer to Japanese Patent Laid-Open No. 2__45). Therefore, it is not preferable to carry out the use of the external one, and (4) the improving agent of the present invention, whereby the (4) effector can be used for the body, and therefore, (4) is improved and the absorbency is also improved. Therefore, the present crusting (there is a decrease in (4) and an increase in (4)) can be said to improve the feed utilization efficiency by using the feed efficiency improving agent of the present invention, and as a result, the amount of milk is also improved. σ (6) Test Example 7 <About decane gas generation amount> The test was carried out according to the above test conditions, and the hood-type breath test apparatus was used to measure the smoldering in the control period i, the test period, and the control period 2 The amount of gas _MI/day) was calculated as the flat value of the control period (i, 2) and the test period, and the results are shown in Table 7. [Table 7] —---—η Control period The amount of decane gas generated (L/DMI/day) 33.4±3.3
甲烧發生量係於對照期卜2之平均值與試驗期之平 中’未確認到有意義的變化。 甲院-般係佔有發生氣體中之3〇%，認為—飼料所攝取 之能量的約5〜8%未被有效利用而以曱烷釋出至大氣中 由上述試驗例2(飼料消化率之提升）等而言， J $月於试驗 100108238The amount of A-burn was based on the average of the control period and the average of the test period. No significant change was confirmed. A hospital-like system accounts for 3〇% of the gas generated, and it is considered that about 5 to 8% of the energy absorbed by the feed is not effectively utilized and released into the atmosphere by decane. The above test example 2 (feed digestibility) Promotion), etc., J $month in test 100108238
S 25 201136528 期（本赉明之飼料利用效率改善劑使用期）中，飼料分解率得 以提升’即使如此，^發生率並未增加，本結果顯示出抑 制了月b里損失並改善飼料利用效率，結果亦顯示改善乳量。 (產業上之可利用性） 本發明由於提升-種發揮可整體改善反_物之特異消 化吸收機構或能量代謝機構之效果的改善劑，故產業上之可 利用性大。 本發明提供一種反离動物之飼料利用效率改善劑與乳量 改善劑’其特徵為並未使用抗生物質或特殊胺基酸等，而改 善反舞動物之瘤胃輯，促進瘤胃微生物之增加及/或活性 化，且提升瘤胃内之丙酸的生成、微生物體蛋白質的生成， 進而’使體时害㈣少且增加有用S，使飼料之消化率、 尤其是含有固形物之飼料的消化率提升，減少 產業上之可利用性大。 、 藉由將本發明之改善劑直接或添加至甸料中而給予，則己 物:瘤胃醱酵並提升瘤胃内之菌體蛋白質合成 用！二 強的氨濃度減少，又’體内有害菌減少幻 ㈣健康且乳量或乳質改善的家畜，㈣ 產有助轉，產業上之可利用性大。 早再者a，藉由將本發明之改善劑直接或添加至飼料中⑽ 可提升飼料利用效率、亦 ，' 損失，故可減少所給予之_==】率並減觸 100108238 杈幵生產性而亦有利於多 26 201136528 濟，此外，可改善家畜之排出物量與質，亦有助於改善環境。 【圖式簡單說明】 圖1為表示試驗例2之飼料消化率變化(提升）的圖。 圖2為表示試驗例3之瘤胃液中之丙酸莫耳比率變化(增 加）的圖。 圖3為表示試驗例4之瘤胃液中之氨態氮濃度變化(減少） 的圖。 圖4為表示試驗例5之瘤胃液中之原生動物數變化(增加) 的圖。 圖5為表示試驗例6之糞中之各種菌數變化（有害菌之減 少及有用菌之增加）的圖。 100108238 27In S 25 201136528 (this period of use of feed utilization efficiency improver), the feed decomposition rate is improved. Even so, the incidence rate does not increase. This result shows that the loss in the month b is suppressed and the feed utilization efficiency is improved. The results also showed an improvement in milk yield. (Industrial Applicability) The present invention has an industrial applicability because it can improve the effect of improving the effect of the specific digestion and absorption mechanism or the energy metabolism mechanism of the anti-object. The present invention provides a feed utilization efficiency improving agent and a milk amount improving agent for a reverse animal, which are characterized in that the anti-biomass or a special amino acid is not used, and the rumen of the anti-dance animal is improved, and the increase of the rumen microorganism is promoted and/ Or activation, and increase the production of propionic acid in the rumen, the production of microbial protein, and then reduce the body's digestibility (four) and increase the useful S, so that the digestibility of the feed, especially the feed containing solids, is increased. To reduce the industrial availability. By administering the improving agent of the present invention directly or in the medicinal material, the rumen is fermented and the bacterial protein synthesis in the rumen is promoted! The second strong ammonia concentration is reduced, and the harmful bacteria in the body are reduced. (4) Livestock with improved milk or milk quality, (4) Production is facilitated, and industrial availability is large. As early as a, by directly or adding the improving agent of the present invention to the feed (10), the feed utilization efficiency and also the loss can be improved, so that the rate of _=== can be reduced and the yield is reduced by 100108238. It is also beneficial to more than 2011, 2011, and to improve the amount and quality of livestock emissions, and also help to improve the environment. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a graph showing changes (elevation) of feed digestibility in Test Example 2. Fig. 2 is a graph showing the change (increased) of the molar ratio of propionate in the rumen fluid of Test Example 3. Fig. 3 is a graph showing a change (decrease) in the concentration of ammonia nitrogen in the rumen fluid of Test Example 4. Fig. 4 is a graph showing the change (increase) in the number of protozoa in the rumen fluid of Test Example 5. Fig. 5 is a graph showing changes in the number of bacteria in the feces of Test Example 6 (reduction of harmful bacteria and increase in useful bacteria). 100108238 27