JP3774199B2 - Antibacterial composition, feed for livestock or poultry using the same, and method for feeding the same - Google Patents

Description

【００１９】
なお、表１の各酸剤の割合は、いずれも１００％純度で示してあるが、試験に用いた各酸剤の純度と性状は、クエン酸＝９９％の粉体、ギ酸＝９９％の液体、乳酸＝９０％の液体、プロピオン酸＝９９％の液体であり、各々の純度差（クエン酸＝１％、ギ酸＝１％、乳酸＝１０％、プロピオン酸＝１％）は、それぞれ、担体である水に含めてある。
【００２０】
（４）試験方法：
上記３種の供試菌のうち、Escherichia coliとSalmonella enteritidisについては、表１の各合剤（合剤１〜合剤６）を濃度を変えて添加したハートインヒュージョン培地にそれぞれ接種し、３５℃で２０時間好気培養して最小発育阻止濃度（ＭＩＣ）を調べた。また、Clostridium perfringens については、表１の各合剤を濃度を変えて添加したＧＡＭ培地に接種し、３５℃で２０時間嫌気培養してＭＩＣを調べた。
【００２１】
（５）試験結果：
合剤１から合剤６について、各菌に対するＭＩＣは表２に示すとおりである。

【００２２】
（６）考察と官能評価：
表２に示すとおり、合剤１から合剤６についてin vitroでおこなった抗菌力試験では、いずれの合剤も同等の抗菌効果があることが確認された。しかし、合剤６については、揮発性が高く、酸臭の強いギ酸とプロピオン酸が併せて２７％も含まれていることから、採用がためらわれた。そのため、合剤１〜合剤６の各サンプルを飼料工場の担当者１０名に提示し、官能評価を依頼した。その結果、やはり、１０名中の９名が合剤６について不採用の意向を示した。この官能評価の結果を考えると、合剤６は、現場でハンドリングに携わる人間への影響が強いので、飼料用の抗菌性組成物としては実用上好ましくないように考えられる。
【００２３】
【試験例２】
＜子豚を用いた選択摂取試験＞
（１）試験方法：
下記の「合剤の製法」にしたがって製した６種の合剤（合剤１〜合剤６）と抗菌力の強さが確認されているギ酸単独の製剤を、それぞれ生後３０日齢の子豚３０頭に３日間給与して選択摂取試験をおこない、家畜の嗜好性への影響を確認した。すなわち、本試験例で供試する合剤１から合剤６は、試験例１の合剤１〜合剤６に対して、水の代わりに同量の二酸化ケイ素を担体として用いた粉末状の合剤である。また、ギ酸単独の製剤は、ギ酸７０％を二酸化ケイ素３０％に噴霧して吸着させたものである。この７種類の合剤又は製剤をそれぞれ０．２％宛、表４に示す子豚用市販飼料に添加したものを、自由に選択して摂取できる状態にして子豚に給与した。
【００２４】
（２）合剤の製法：
各合剤は、以下の方法によって製した。
《合剤１》
クエン酸６６７ｇ、ギ酸５１ｇ、乳酸２２ｇ、二酸化ケイ素２６０ｇを正確に秤量し、最初にクエン酸と二酸化ケイ素の均一な粉末を作り、これにギ酸と乳酸の均一な混合液を噴霧し、性状が安定するまで静置した後、攪拌して均一な粉末状の合剤１０００ｋｇを得た。
《合剤２》
クエン酸６６７ｇ、ギ酸５１ｇ、プロピオン酸２０ｇ、二酸化ケイ素２６２ｇを正確に秤量し、最初にクエン酸と二酸化ケイ素の均一な粉末を作り、これにギ酸とプロピオン酸の均一な混合液を噴霧し、性状が安定するまで静置した後、攪拌して均一な粉末状の合剤１０００ｋｇを得た。
《合剤３》
クエン酸４６５ｇ、ギ酸１５２ｇ、乳酸１３３ｇ、二酸化ケイ素２５０ｇを正確に秤量し、最初にクエン酸と二酸化ケイ素の均一な粉末を作り、これにギ酸と乳酸の混合液を噴霧し、性状が安定するまで静置した後、攪拌して均一な粉末状の合剤１０００ｋｇを得た。
《合剤４》
クエン酸５０５ｇ、ギ酸５１ｇ、プロピオン酸１２１ｇ、二酸化ケイ素３２３ｇを正確に秤量し、最初にクエン酸と二酸化ケイ素の均一な粉末を作り、これにギ酸とプロピオン酸の均一な混合液を噴霧し、性状が安定するまで静置した後、攪拌して均一な粉末状の合剤１０００ｋｇを得た。
《合剤５》
クエン酸４６５ｇ、ギ酸１０１ｇ、乳酸６７ｇ、プロピオン酸６１ｇ、二酸化ケイ素３０６ｇを正確に秤量し、最初にクエン酸と二酸化ケイ素の均一な粉末を作り、これにギ酸と乳酸とプロピオン酸の均一な混合液を噴霧し、性状が安定するまで静置した後、攪拌して均一な粉末状の合剤１０００ｋｇを得た。
《合剤６》
クエン酸４６５ｇ、ギ酸１５２ｇ、プロピオン酸１２１ｇ、二酸化ケイ素２６２ｇを正確に秤量し、最初にクエン酸と二酸化ケイ素の均一な粉末を作り、これにギ酸とプロピオン酸の均一な混合液を噴霧して、性状が安定するまで静置した後、攪拌して均一な粉末状の合剤１０００ｋｇを得た。
【００２５】
（３）試験結果：
試験の結果、表３に示す成績が得られた。
【表３】

【００２６】
（４）考察：
表３から、子豚の嗜好について、合剤１〜合剤５の間には、極端な差異は見られなかった。なお、合剤１〜合剤５の抗菌力は、先のin vitroでの試験（試験例１）でも同等であることが確認されている。しかし、ギ酸単独の製剤と合剤６については、やはり、子豚の嗜好に適していないことが確認された。ギ酸やプロピオン酸の抗菌力の強さは広く知られているところであるが、家畜にも明確な拒絶反応が確認されたので、先の官能評価と併せて、合剤６は、家畜又は家禽用の抗菌性組成物としては限界を越えていると判断し、以降は、合剤１から合剤５までにしぼってさらに動物試験を続けることにした。
なお、供試した市販飼料の配合は表４に示すとおりである。
【００２７】
【表４】

【００２８】
【試験例３】
＜合剤の保存試験＞
（１）試験方法：
試験例２の製法で作成した合剤１〜合剤６をそれぞれ２か月間常温で保管し、保存劣化の有無を確認した。
（２）試験結果と考察：
２か月の常温保管後、合剤３と合剤６には、若干の固まりが発見された。この固まりは手で砕ける程度の軽微なもので、使用上、実質的に問題が生じることはなかった。しかし、その原因は、合剤３と合剤６は、ギ酸、乳酸、プロピオン酸といった液状の酸剤の含量が多いので、そのことに起因するものと推察された。（３）合剤７の試験：
そこで、さらに、クエン酸４０４ｇ（４０％）、ギ酸１０１ｇ（１０％）、乳酸１１１ｇ（１０％）、プロピオン酸１０１ｇ（１０％）、担体として二酸化ケイ素２８３ｇを用い、試験例２と同じ製法で「合剤７」を作成し、この合剤７を２か月間常温で保管して保存劣化の有無を確認した。
その結果、合剤７には、手では砕けない大きな固まりが発見された。このことから、合剤７（液状の酸剤の合計量が３１．３％）は、抗菌力はともかく、製剤化には不適であり、合剤３（液状の酸剤の合計量が２８．５％）と合剤６（液状の酸剤の合計量は２７．３％）が、実用上、抗菌性組成物として用いる限界であると判断された。なお、合剤７は、試験例２の結果を考慮すると、子豚の嗜好に適さない上、飼料工場や畜産現場でのハンドリングに耐え得るものではない。また、先記のとおり、合剤６も同様に判断された。
以上のことから、結論として「合剤１から合剤５までの酸剤の組み合わせが」が抗菌性組成物として実用性のある範囲であると判断された。
【００２９】
【試験例４】
＜採卵鶏と子牛の糞中排菌測定＞
（１）試験方法と試験結果：
試験例２の製法で作成した合剤２を０．０５％添加した市販飼料を３００日齢の採卵鶏３０羽と４３日齢の子牛１５頭にそれぞれ連続して７日間給与した。その結果、どちらも、飼料摂取量に特段の差異は見られなかった。しかし、この試験区と同区域で飼育されていた採卵鶏と子牛の「合剤無添加飼料給与群」では、どちらも、数羽ないし数頭が大腸菌に起因する下痢を発症した。一方、「合剤２添加飼料給与群」では、採卵鶏も子牛も、下痢の発生自体が圧倒的に少なく、細菌性の下痢を発症したものは全く観察されなかった。そして、試験終了直後に採取した「合剤２添加飼料給与群」の採卵鶏と子牛の新鮮糞を検査したところ、
Clostridium perfringens 、Enterotoxigenic 、Enteropathogenic Escherichia coli 、Salmonella enteritidis、Salmonella infantis は、いずれも全く検出されなかった。
供試した市販飼料の配合割合は、表５と表６に示すとおりである。
【００３０】
（２）採卵鶏用市販飼料の配合割合：
【表５】

【００３１】
（３）子牛用市販飼料の配合割合：
【表６】

【００３２】
【試験例５】
＜肥育豚の糞中排菌測定＞
（１）試験方法と試験結果：
試験例２の製法で作成した合剤１を０．０５％添加した市販飼料を８２日齢の肥育豚３０頭に連続して７日間給与した。その結果、飼料摂取量に特段の差異は見られなかった。しかし、この試験区と同区域で飼育されていた肥育豚の「合剤無添加飼料給与群」では、数頭が大腸菌に起因する下痢を発症した。一方、「合剤１添加飼料給与群」の肥育豚では、下痢の発生自体が圧倒的に少なく、細菌性の下痢を発症したものは全く観察されなかった。そして、試験終了直後に採取した「合剤２添加飼料給与群」の肥育豚の新鮮糞を検査したところ、クロストリジウム・パーフリンゲンス、大腸菌、サルモネラ菌は、いずれも全く検出されなかった。供試した市販飼料の配合割合は、表７に示すとおりである。
【００３３】
（２）肥育豚用市販飼料（子豚育成用配合飼料）の配合割合：
【表７】

【００３４】
試験例１〜試験例５によって、家畜又は家禽の嗜好を損なわず、しかも主要な細菌性疾病の予防に有効な抗菌性組成物として好ましい合剤の組成が確認されたので、次に、試験例６と試験例７をもって、これらの抗菌性組成物（合剤）を添加した飼料の家畜や家禽の発育に対する影響を確認することにした。
【００３５】
【試験例６】
＜ブロイラー発育試験＞
（１）試験方法：
２１日齢のブロイラー１５０羽を５つの試験区と１つの対照区に分けて、試験区のブロイラーにはそれぞれ表８に示す試験区飼料（ブロイラー用配合飼料に合剤１〜合剤５を各０．３％添加した５通りの飼料）を、また、対照区のブロイラーには、表８に示す対照区飼料（合剤無添加のブロイラー用配合飼料）を、いずれも２８日間給与して、それぞれの発育への影響を確認した。
【００３６】
（２）試験区飼料と対照区飼料：
試験区飼料に添加した５通りの合剤（合剤１〜合剤５）は、いずれも、試験例２の「合剤の製法」において、二酸化ケイ素の代わりに小麦粉を用いて製したものである。（例えば、合剤１であれば、クエン酸６６７ｇと小麦粉２６０ｇの均一な粉末を作り、これにギ酸５１ｇと乳酸２２ｇの均一な混合液を噴霧し、性状が安定するまで静置した後、攪拌して均一化した粉末状の合剤である。）
本試験例で用いる５通りの飼料には、表８に示すとおり、上記の製法で作成した合剤１〜合剤５を各０．３％添加してある。
また、本試験例で用いる対照区飼料は、表８に示すように、これら合剤は添加していないが、その他の原料の配合を試験区飼料とほとんど同じ割合にした飼料である。
【００３７】
（３）供試したブロイラー用配合飼料：
【表８】

【００３８】
（４）ブロイラーの発育成績
【表９】

【００３９】
（５）考察：
表９によれば、合剤１〜合剤５をそれぞれ０．３％添加した試験区飼料では、５通りの試験区飼料の全てが対照区飼料に比べて、ブロイラーの飼料摂取量がわずかに向上しており、また、日増体も試験区飼料給与の方が高くなっている。したがって、本発明に係る抗菌性組成物は、合剤１〜合剤５のいずれについても細菌性疾病の予防剤として有効な量を飼料に添加して給与したにもかかわらず、ブロイラーの嗜好や発育には何も悪い影響が生じないことが確認された。
【００４０】
【試験例７】
＜離乳子豚の発育試験＞
（１）試験方法：
試験例６と同様の試験を離乳子豚についても実施した。すなわち、２１日齢の離乳子豚３０頭を５つの試験区と１つの対照区に分けて、試験区の子豚にはそれぞれ表１０に示す試験区飼料（子豚用配合飼料に合剤１〜合剤５を各０．３％添加した５通りの飼料）を、また、対照区の子豚には表１０に示す対照区飼料（合剤無添加の子豚用配合飼料）を、いずれも１４日間給与して、それぞれの発育への影響を確認した。
【００４１】
（２）試験区飼料と対照区飼料：
試験区飼料に添加した５通りの合剤（合剤１〜合剤５）は、いずれも、試験例２の「合剤の製法」において、二酸化ケイ素の代わりに小麦粉を用いて製したものである。（例えば、合剤２であれば、クエン酸６６７ｇと小麦粉２６２ｇの均一な粉末を作り、この粉末にギ酸５１ｇとプロピオン酸２０ｇの均一な混合液を噴霧し、性状が安定するまで静置した後、攪拌して均一化した粉末状の合剤である。）
本試験例で用いる５通りの飼料には、表１０に示すとおり、上記の製法で作成した合剤１〜合剤５を各０．３％添加してある。
また、本試験例で用いる対照区飼料は、表１０に示すように、これら合剤は添加していないが、その他の原料の配合を試験区飼料とほとんど同じ割合にした飼料である。
【００４２】
（３）供試した子豚用配合飼料：
【表１０】

【００４３】
（４）離乳子豚の発育成績
【表１１】

【００４４】
（５）考察：
表１１によれば、合剤１〜合剤５をそれぞれ０．３％添加した試験区飼料において、５通りの試験区飼料の全てが対照区飼料に比べて、子豚の飼料摂取量が向上しており、また、日増体も試験区飼料給与の方が高くなっている。さらに、試験区飼料では軟便の発生率が大幅に減少している。したがって、本発明に係る抗菌性組成物は、合剤１〜合剤５のいずれについても細菌性疾病の予防剤として有効な量を飼料に添加して給与したにもかかわらず、離乳子豚の嗜好や発育には何も悪い影響が生じないことが確認された。
【００４５】
以上の試験結果から、家畜及び家禽の主要な細菌性疾病の予防に有効であり、かつ家畜及び家禽の嗜好を損なわず、発育を阻害することがない抗菌性組成物は「表１に示す合剤１〜合剤５の各酸剤の組み合わせ」であることが確認された。したがって、本発明に係る抗菌性組成物としては、「クエン酸４６〜６６％、ギ酸５〜１５％、乳酸及び／又はプロピオン酸２〜１２％を主材とする合剤」が最適であることが理解される。また、本発明に係る家畜又は家禽用飼料としては、上記の合剤１〜合剤５を抗菌性組成物として０．０５％（試験例４）から０．２％（試験例２）ないし０．３％（試験例６及び試験例７）を含有させることが適切であることが理解される。
【００４６】
【発明の効果】
以上、詳しく説明のとおり、本発明に係る家畜又は家禽用の抗菌性組成物は、特定配合量のクエン酸、ギ酸、乳酸及び／又はプロピオン酸を担体に添加したものであるから、抗生物質のような耐性菌発現のおそれがない。それでいて、クロストリジウム・パーフリンゲンス症、大腸菌症及びサルモネラ菌症のいずれに対しても十分な予防力を有する上、家畜や家禽の成長に悪い影響を与えるおそれがない。しかも、家畜にも家禽にも効果があるので、両者共用の予防剤として用いることができる。
【００４７】
また、本発明に係る家畜又は家禽用飼料は、上記の抗菌性組成物を通常の飼料へ少量だけ添加したものであるから、現場においてハンドリングしやすく、しかも、有機酸を添加しただけであるから家畜又は家禽の成長や嗜好に悪い影響を与えるおそれがない。その上、家畜又は家禽にとってきわめて摂取しやすいものである。
本発明に係る家畜又は家禽用飼料は、それでいて、抗生物質を添加した飼料と同等に、クロストリジウム・パーフリンゲンス症、大腸菌症及びサルモネラ菌症のいずれに対しても十分な予防力を有する。しかも、同じ飼料を家畜及び家禽に共用できる。また、本発明に係る家畜又は家禽用飼料の給与方法は、きわめて簡単であり、家畜又は家禽の嗜好に影響を与えないので、きわめて給与しやすい。以上のとおり、本発明は、家畜及び家禽に共通の主要な細菌性疾病に対して、きわめて有用な予防手段を提供するものであり、飼料業界にとっても、また畜産業界にとっても大いに有意義なものである。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to an antibacterial composition, livestock or poultry feed using the same, and a method for feeding the same. Specifically, antibacterial compositions effective for the prevention of major bacterial diseases common to livestock or poultry, livestock or poultry feeds that are easy to ingest using the antibacterial compositions, and feed the livestock or poultry It relates to a preferred method.
[0002]
[Prior art]
[Patent Document 1]
JP 2001-238609 A [Patent Document 2]
JP 2002-12505 A [Patent Document 3]
JP-A-10-215790 gazette
Traditionally, the prevention of disease in livestock and poultry derived from bacteria has tended to depend on antibiotics, and concerns that the addition of antibiotics induces the development of resistant bacteria and puts the human body at risk. It is regarded as a problem. Therefore, in the livestock industry, development of antibacterial materials that can replace antibiotics is required.
[0004]
On the other hand, the use of organic acids for the control of bacterial diseases in livestock and poultry is a technique that has been widely discussed, and several patent applications have been found. For example, JP-A-2001-238609 discloses a feed for combating Salmonella to which mannose-containing copra meal and organic acid are added, and as the organic acid, acetic acid, formic acid, propionic acid, lactic acid, fumaric acid, or citric acid can be used. Are listed. Japanese Patent Application Laid-Open No. 2002-12505 discloses a Salmonella and / or Escherichia coli contamination-preventing feed in which solid formic acid supported on an inorganic substance such as hydrous silicon dioxide and vermiculite is mixed with the feed. Furthermore, JP-A-10-215790 discloses a poultry feed supplemented with oligosaccharides and acidifying materials for reducing bacterial infection such as Salmonella, and as acidifying materials, propionic acid, formic acid or Citric acid is mentioned. Thus, organic acids such as propionic acid, formic acid, acetic acid, lactic acid, fumaric acid and citric acid are known to be used in livestock or poultry feeds as preventive or therapeutic agents for bacterial infections. As described in the above publication, all of the preventive agents for bacterial infectious diseases using organic acids have a complicated structure, and it is necessary to increase the amount of organic acid added to the feed in order to increase the preventive effect. Therefore, it does not match the taste of livestock and poultry and is difficult to take.
[0005]
In view of such a situation, the present inventors have continued research on an antibacterial material that can be substituted for antibiotics using an organic acid as a main material and combine the organic acid with a specific blending ratio. Synergistic action of organic acids occurs, and it is effective against Clostridium perfringens known as the cause of fulminant enteritis as well as Salmonella and Escherichia coli. It was found that an antibacterial composition that can be easily ingested by livestock and poultry can be developed because the amount is small, and further, a confirmation test as livestock feed and poultry feed was conducted, and the present invention was completed.
[0006]
[Problems to be solved by the invention]
That is, the present invention is composed of an organic acid as a main material and can be replaced with antibiotics, and can be easily consumed by livestock and poultry because the amount of feed can be reduced. New antibacterial composition that does not adversely affect growth, can be shared by livestock and poultry, is easy to handle in the field, and is effective in preventing Clostridium perfringens, colitis and Salmonella in livestock and poultry It is an object of the present invention to provide a livestock or poultry feed that is useful for the prevention of these bacterial diseases, and an effective method for feeding the livestock or poultry feed using the product.
[0007]
[Means for Solving the Problems]
Among the present inventions for solving the above-mentioned problems, the invention described in claim 1 includes citric acid 46 to 66%, formic acid 5 to 15%, lactic acid and / or propionic acid 2 to 12%, and carriers 27 to 33. It is an antibacterial composition effective for the prevention of Clostridium perfringens, colibacillosis and Salmonella in domestic animals or poultry.
[0008]
Moreover, invention described in Claim 2 among the present invention is a feed for livestock or poultry containing 0.05 to 0.3% of the antibacterial composition described in Claim 1.
[0009]
Moreover, the invention described in claim 3 of the present invention is that the livestock or poultry feed according to claim 2 is fed to livestock or poultry for 7 days or more, and the livestock or poultry clostridial perfringens disease, Escherichia coli This is a method for preventing the disease and Salmonella.
[0010]
Hereinafter, the present invention will be described in more detail.
In the present invention, “%” indicates “% by weight” unless otherwise specified. In the present invention, a mixture of organic acids added with a carrier or a mixture of organic acids added to a carrier is referred to as a “mix”, and each organic acid constituting the mixture is also referred to as an “acid”. Called.
In the present invention, the meaning of “livestock or poultry” means either livestock or poultry or both. For example, “livestock or poultry feed” means either feed for livestock or poultry feed and feed that can be fed to both livestock and poultry.
[0011]
The antibacterial composition according to the present invention is a combination of organic acids consisting of citric acid, formic acid, lactic acid and / or propionic acid in a specific ratio, which is added to a powdery or liquid carrier, By adding, it is in the state which is easy to mix uniformly in feed.
[0012]
In the present invention, it is preferable to use powders such as silicon dioxide and wheat flour as the carrier of each acid agent. The carrier used in the present invention is not particularly limited as long as it is not harmful to livestock or poultry as long as it can be mixed with feed by adding each of the above acid agents.
[0013]
As the antibacterial composition according to the present invention, it is preferable to use an antibacterial composition prepared by spraying an organic acid of a specific formulation onto a powder carrier and uniformly adsorbing it, and producing it as a powder mixture.
The antibacterial composition according to the present invention is effective only by adding a small amount to livestock or poultry feed. Moreover, the antibacterial composition which concerns on this invention can share the mixture of the same composition for the feed for livestock, and the feed for poultry. In general, it is preferable to add 0.05% to 0.3% with respect to normal livestock feed or normal poultry feed.
[0014]
That is, as shown in the following test examples, the livestock or poultry feed according to the present invention can exhibit a sufficient preventive effect against major bacterial diseases even if the addition amount of the antibacterial composition is small. Therefore, the feed for livestock or poultry according to the present invention can be supplied very easily without the risk of impairing the taste of livestock or poultry because the amount of the antibacterial composition added is small. In addition, handling at feed factories and livestock production is easy. Further, the feed for livestock or poultry according to the present invention has a large preventive effect when fed to young animals or chickens that are not physically strong and susceptible to bacterial diseases, but is not limited thereto. It also has a sufficient preventive effect on bacterial diseases.
[0015]
If the livestock or poultry feed according to the present invention is fed to poultry or livestock for approximately 7 days, the above-mentioned preventive effect against the main bacterial diseases can be obtained. In addition, the feeding method can be either constant feeding or restricted feeding.
[0016]
Hereinafter, the present invention will be described in more detail with test examples.
In each of the following test examples, each mixture to be used for the test is referred to as “mixture 1” or the like, but the same “mixture 1” has different carriers. That is, “Compound 1” in Test Example 1 is a liquid mixture (66% citric acid , 5% formic acid, 2% lactic acid, and water as a carrier) shown in the “Compound 1” column of Table 1. Are added and mixed). In addition, “Compound 1” of Test Examples 2 to 5 is a powdered mixture (66% citric acid) manufactured according to the section of “Compound 1” of “Production Method” shown in Test Example 2. , 5% formic acid, 2% lactic acid and silicon dioxide as a carrier). Further, “Compound 1” of Test Example 6 and Test Example 7 is a powdered mixture (citric acid) using wheat flour instead of silicon dioxide as a carrier for the mixture 1 of Test Examples 2 to 5. 66%, formic acid 5%, and lactic acid 2% as main materials and flour as a carrier).
[0017]
[Test Example 1]
<In vitro antibacterial activity test>
(1) Test bacteria:
Clostridium perfringens, Escherichia coli, Salmonella enteritidis
(2) Number of inoculated bacteria: 10 ⁸ each [0018]
(3) The composition of the tested antibacterial composition (mixture) is as shown in Table 1.

[0019]
In addition, although the ratio of each acid agent of Table 1 is shown in 100% purity, the purity and properties of each acid agent used in the test are citric acid = 99% powder, formic acid = 99%. Liquid, lactic acid = 90% liquid, propionic acid = 99% liquid, and the respective purity differences (citric acid = 1%, formic acid = 1%, lactic acid = 10%, propionic acid = 1%) It is included in the carrier water.
[0020]
(4) Test method:
Among the above three types of test bacteria, Escherichia coli and Salmonella enteritidis were each inoculated into a heart infusion medium to which each of the combinations (mixtures 1 to 6) in Table 1 was added at different concentrations, and 35 The minimum growth inhibitory concentration (MIC) was examined by aerobic culture at 20 ° C. for 20 hours. Also, the Clostridium perfri ng ens, each mixture of Table 1 was inoculated to GAM medium supplemented with different concentrations were examined MIC was 20 hours anaerobic incubation at 35 ° C..
[0021]
(5) Test results:
Regarding mixture 1 to mixture 6, the MIC for each bacterium is as shown in Table 2.

[0022]
(6) Discussion and sensory evaluation:
As shown in Table 2, in the antibacterial activity test conducted in vitro on the mixture 1 to the mixture 6, it was confirmed that all the combinations had an equivalent antibacterial effect. However, the mixture 6 was hesitant to be adopted because it contains 27% of formic acid and propionic acid which are highly volatile and have strong acid odor. Therefore, each sample of the mixture 1 to the mixture 6 was presented to 10 persons in charge of the feed factory and requested for sensory evaluation. As a result, 9 out of 10 people indicated that the mixture 6 was not adopted. Considering the result of this sensory evaluation, the mixture 6 has a strong influence on humans who are involved in handling in the field, so it is considered to be unpractically preferable as an antibacterial composition for feed.
[0023]
[Test Example 2]
<Selective intake test using piglets>
(1) Test method:
Six types of combinations (mixtures 1 to 6) manufactured according to the following “mixture manufacturing method” and formic acid-only preparations that have been confirmed to have strong antibacterial activity are each a 30-day-old child. 30 pigs were fed for 3 days and a selective intake test was conducted to confirm the influence on the preference of livestock. That is, the mixture 1 to the mixture 6 to be tested in this test example are in the form of powder using the same amount of silicon dioxide as a carrier instead of water with respect to the mixture 1 to the mixture 6 of the test example 1. It is a mixture. Further, the preparation of formic acid alone is obtained by spraying 70% formic acid onto 30% silicon dioxide and adsorbing it. These 7 types of combinations or preparations were each fed to 0.2% and added to the commercial feed for piglets shown in Table 4 to be freely selected and fed to the piglets.
[0024]
(2) Mixture manufacturing method:
Each mixture was produced by the following method.
<< mixture 1 >>
Accurately weigh 667 g of citric acid, 51 g of formic acid, 22 g of lactic acid, and 260 g of silicon dioxide, first make a uniform powder of citric acid and silicon dioxide, spray a uniform mixture of formic acid and lactic acid on this, and stable properties Then, the mixture was stirred and 1000 kg of a uniform powdery mixture was obtained.
<< mixture 2 >>
Accurately weigh 667 g of citric acid, 51 g of formic acid, 20 g of propionic acid, and 262 g of silicon dioxide, first make a uniform powder of citric acid and silicon dioxide, spray a uniform mixture of formic acid and propionic acid on this, The mixture was allowed to stand until it became stable, and then stirred to obtain 1000 kg of a uniform powdery mixture.
<< mixture 3 >>
Accurately weigh 465 g of citric acid, 152 g of formic acid, 133 g of lactic acid, and 250 g of silicon dioxide, first make a uniform powder of citric acid and silicon dioxide, spray a mixture of formic acid and lactic acid on this, until the properties are stable After standing, the mixture was stirred to obtain 1000 kg of a uniform powdery mixture.
<< mixture 4 >>
Accurately weigh 505 g of citric acid, 51 g of formic acid, 121 g of propionic acid, and 323 g of silicon dioxide, first make a uniform powder of citric acid and silicon dioxide, spray a uniform mixture of formic acid and propionic acid on this, The mixture was allowed to stand until it became stable, and then stirred to obtain 1000 kg of a uniform powdery mixture.
<< mixture 5 >>
465 g of citric acid, 101 g of formic acid, 67 g of lactic acid, 61 g of propionic acid, and 306 g of silicon dioxide are accurately weighed to make a uniform powder of citric acid and silicon dioxide first, and a uniform mixture of formic acid, lactic acid and propionic acid Was sprayed and allowed to stand until the properties were stable, and then stirred to obtain 1000 kg of a uniform powdery mixture.
<< mixture 6 >>
Accurately weigh 465 g of citric acid, 152 g of formic acid, 121 g of propionic acid, and 262 g of silicon dioxide, first make a uniform powder of citric acid and silicon dioxide, and spray this with a uniform mixture of formic acid and propionic acid, The mixture was allowed to stand until the properties became stable, and then stirred to obtain 1000 kg of a uniform powdery mixture.
[0025]
(3) Test results:
As a result of the test, the results shown in Table 3 were obtained.
[Table 3]

[0026]
(4) Consideration:
From Table 3, about the preference of a piglet, the extreme difference was not seen between the mixture 1 to the mixture 5. In addition, it was confirmed that the antibacterial activity of the mixture 1 to the mixture 5 is equivalent in the previous in vitro test (Test Example 1). However, it was confirmed that the formulation of formic acid alone and combination 6 were not suitable for the preference of piglets. The strength of antibacterial activity of formic acid and propionic acid is widely known, but since a clear rejection reaction was confirmed in livestock, combined with the previous sensory evaluation, mixture 6 was used for livestock or poultry. It was determined that the antibacterial composition was over the limit, and thereafter, it was decided to continue further animal testing by narrowing down from mixture 1 to mixture 5.
In addition, the composition of the commercial feed used was as shown in Table 4.
[0027]
[Table 4]

[0028]
[Test Example 3]
<Preservation test of mixture>
(1) Test method:
Mixtures 1 to 6 prepared by the production method of Test Example 2 were each stored at room temperature for 2 months, and the presence or absence of storage deterioration was confirmed.
(2) Test results and discussion:
After storage at room temperature for 2 months, some solids were found in mixture 3 and mixture 6. This lump was so small that it could be crushed by hand, and practically no problem occurred. However, the cause was presumed to be due to the fact that the mixture 3 and the mixture 6 contained a large amount of liquid acid agents such as formic acid, lactic acid, and propionic acid. (3) Test of Mixture 7:
Accordingly, 404 g (40%) of citric acid, 101 g (10%) of formic acid, 111 g (10%) of lactic acid, 101 g (10%) of propionic acid, and 283 g of silicon dioxide as a carrier were used. A “mixture 7” was prepared, and the mixture 7 was stored at room temperature for 2 months to check for storage deterioration.
As a result, a large mass of the mixture 7 that could not be broken by hand was found. From this, mixture 7 (total amount of liquid acid agent is 31.3%) is unsuitable for formulation, apart from antibacterial activity, and mixture 3 (total amount of liquid acid agent is 28. 5%) and Mixture 6 (the total amount of the liquid acid agent was 27.3%) were determined to be practical limits for use as an antibacterial composition. In addition, when the result of Test Example 2 is taken into consideration, the mixture 7 is not suitable for the preference of piglets and cannot withstand handling at a feed factory or livestock production site. Further, as described above, the mixture 6 was also judged in the same manner.
From the above, as a conclusion, it was determined that “the combination of acid agents from mixture 1 to mixture 5” is in a practical range as an antibacterial composition.
[0029]
[Test Example 4]
<Measurement of fecal excretion of hens and calves>
(1) Test method and test results:
A commercial feed supplemented with 0.05% of the mixture 2 prepared by the production method of Test Example 2 was fed continuously to 30 300-day-old egg-laying hens and 15 43-day-old calves for 7 days. As a result, in both cases, there was no particular difference in feed intake. However, in the “fed group without supplements” of egg-laying hens and calves bred in the same area as this test area, several or several animals developed diarrhea caused by E. coli. On the other hand, in the “Feeding group with 2 supplements”, the occurrence of diarrhea was overwhelmingly small in egg-laying chickens and calves, and no bacterial diarrhea was observed at all. And after examining the fresh feces of the hens and calves of the “mixed feed supplemented feed group” collected immediately after the test,
None of Clostridium perfringens, Enterotoxigenic, Enteropathogenic Escherichia coli, Salmonella enteritidis, or Salmonella infantis were detected.
The blending ratios of the commercial feeds that were tested are as shown in Tables 5 and 6.
[0030]
(2) Mixing ratio of commercial feed for egg laying hens:
[Table 5]

[0031]
(3) Mixing ratio of commercial feed for calves:
[Table 6]

[0032]
[Test Example 5]
<Measurement of fecal excretion in fattening pigs>
(1) Test method and test results:
A commercial feed supplemented with 0.05% of mixture 1 prepared by the production method of Test Example 2 was fed continuously to 30 82-day-old fattening pigs for 7 days. As a result, there was no particular difference in feed intake. However, in the “pigment-free feed group” of fattening pigs bred in the same area as this test area, several dogs developed diarrhea caused by E. coli. On the other hand, in the fattening pigs in the “group 1 supplemented feed feeding group”, the occurrence of diarrhea was overwhelmingly small and no bacterial diarrhea was observed at all. And when the fresh feces of the fattening pigs of the “group 2 supplement feed feeding group” collected immediately after the test was examined, none of Clostridium perfringens, Escherichia coli and Salmonella was detected. Table 7 shows the blending ratio of the commercial feed that was tested.
[0033]
(2) Mixing ratio of commercial feed for fattening pigs (formulated feed for raising piglets):
[Table 7]

[0034]
Since Test Example 1 to Test Example 5 confirmed the composition of a preferable mixture as an antibacterial composition effective in preventing major bacterial diseases without impairing the taste of livestock or poultry, the following test example With 6 and Test Example 7, it was decided to confirm the influence of feed containing these antibacterial compositions (mixtures) on the growth of livestock and poultry.
[0035]
[Test Example 6]
<Brother growth test>
(1) Test method:
Dividing 150 21-day-old broilers into 5 test groups and 1 control group, each broiler in the test group was fed with test group feeds shown in Table 8 (mixed feeds for broilers 1 to 5). 5 feeds with 0.3% added), and the broilers in the control group were fed the control group feeds shown in Table 8 (mixed feed for broilers with no mixture added) for 28 days, The influence on each development was confirmed.
[0036]
(2) Test plot and control plot feed:
The five combinations (mixtures 1 to 5) added to the test zone feed were all made using wheat flour instead of silicon dioxide in Test Method 2 “Production of mixture”. is there. (For example, in the case of the mixture 1, a uniform powder of 667 g of citric acid and 260 g of flour is prepared, and a uniform mixed solution of 51 g of formic acid and 22 g of lactic acid is sprayed on the mixture, and the mixture is allowed to stand until the properties are stabilized, and then stirred. It is a powdered mixture that has been homogenized.)
As shown in Table 8, 0.3% of each of the mixture 1 to the mixture 5 prepared by the above production method is added to the five types of feed used in this test example.
In addition, as shown in Table 8, the control group feed used in this test example is a feed in which these materials are not added, but other raw materials are mixed in almost the same proportion as the test group feed.
[0037]
(3) Formulated feed for broiler used:
[Table 8]

[0038]
(4) Growth performance of broilers [Table 9]

[0039]
(5) Consideration:
According to Table 9, in the test group feed to which 0.3% of each of the mixture 1 to the mixture 5 was added, the broiler feed intake was slightly lower in all of the five test group feeds compared to the control group feed. In addition, daily gain is also higher for feed in the test area. Therefore, the antibacterial composition according to the present invention has a broiler taste or a combination of any of the mixture 1 to the mixture 5, even though the amount effective as a preventive agent for bacterial diseases is added to the feed and fed. It was confirmed that there was no negative effect on development.
[0040]
[Test Example 7]
<Development test of weaned piglets>
(1) Test method:
A test similar to Test Example 6 was also performed on weaned piglets. That is, 30 21-day-old weaned piglets were divided into five test plots and one control plot, and the test plot feeds shown in Table 10 (mixed feed 1 for piglets) ~ 5 kinds of feeds each containing 0.3% of mixture 5), and for control piglets, the control group feeds shown in Table 10 (mixed feed for piglets with no mixture added) Also paid for 14 days to confirm their effects on growth.
[0041]
(2) Test plot and control plot feed:
The five combinations (mixtures 1 to 5) added to the test zone feed were all made using wheat flour instead of silicon dioxide in Test Method 2 “Production of mixture”. is there. (For example, in the case of the mixture 2, after making a uniform powder of 667 g of citric acid and 262 g of wheat flour, spraying a uniform mixed solution of 51 g of formic acid and 20 g of propionic acid on this powder and leaving it until the properties are stabilized. This is a powder mixture that has been homogenized by stirring.)
As shown in Table 10, 0.3% of each of the mixture 1 to the mixture 5 prepared by the above-described production method is added to the five types of feed used in this test example.
In addition, as shown in Table 10, the control group feed used in this test example is a feed in which these materials are not added, but other raw materials are mixed in almost the same proportion as the test group feed.
[0042]
(3) Formulated feed for piglets tested:
[Table 10]

[0043]
(4) Growth performance of weaned piglets [Table 11]

[0044]
(5) Consideration:
According to Table 11, in the test plot feed to which 0.3% of each of mixture 1 to mix 5 was added, the feed intake of the piglet was improved in all of the five test plot feeds compared to the control plot feed In addition, the daily gain is also higher for the feed in the test area. In addition, the incidence of loose stool is greatly reduced in the test area feed. Therefore, the antibacterial composition according to the present invention is a combination of the weaned piglets in spite of the addition of an effective amount as a preventive agent for bacterial diseases to the feed for any of the mixture 1 to the mixture 5. It was confirmed that there was no adverse effect on taste and development.
[0045]
From the above test results, it is effective in preventing the major bacterial diseases of livestock and poultry, or One livestock and without impairing the taste of poultry, the antimicrobial composition does not inhibit the growth in "Table 1 It was confirmed that it is a “combination of each acid agent of mixture 1 to mixture 5 ”. Therefore, as the antibacterial composition according to the present invention, “a mixture containing 46 to 66% citric acid, 5 to 15% formic acid, and 2 to 12% lactic acid and / or propionic acid” is optimal. Is understood. In addition, as feed for livestock or poultry according to the present invention, 0.05% (Test Example 4) to 0.2% (Test Example 2) to 0 of the above mixture 1 to 5 as an antibacterial composition It is understood that it is appropriate to contain 3% (Test Example 6 and Test Example 7).
[0046]
【The invention's effect】
As described above in detail, the antibacterial composition for livestock or poultry according to the present invention is obtained by adding a specific amount of citric acid, formic acid, lactic acid and / or propionic acid to a carrier. There is no fear of developing such resistant bacteria. Nevertheless, it has sufficient preventive power against any of Clostridium perfringens, Escherichia coli, and Salmonella, and there is no risk of adversely affecting the growth of livestock and poultry. Moreover, since it is effective for both livestock and poultry, it can be used as a preventive agent shared by both.
[0047]
In addition, the feed for livestock or poultry according to the present invention is obtained by adding a small amount of the above-mentioned antibacterial composition to normal feed, so that it is easy to handle in the field and only an organic acid is added. There is no risk of adversely affecting the growth or taste of livestock or poultry. In addition, it is very easy for livestock or poultry.
The feed for livestock or poultry according to the present invention still has sufficient preventive power against any of Clostridium perfringens, Escherichia coli and Salmonella as well as a feed to which antibiotics are added. Moreover, the same feed can be shared for livestock and poultry. In addition, the feed method for livestock or poultry feed according to the present invention is very simple and does not affect the taste of livestock or poultry, so it is very easy to feed. As described above, the present invention provides a very useful preventive measure against major bacterial diseases common to livestock and poultry, and is highly significant for both the feed industry and the livestock industry. is there.

Claims (3)

It consists of 46 to 66% citric acid, 5 to 15% formic acid, 2 to 12% lactic acid and / or propionic acid, and 27 to 33% carrier. Prevention of Clostridium perfringens disease, colibacillosis and salmonella in domestic animals or poultry Effective antibacterial composition. A feed for livestock or poultry containing 0.05 to 0.3% of the antibacterial composition according to claim 1. A method of feeding livestock or poultry feed according to claim 2 to livestock or poultry for 7 days or longer to prevent Clostridium perfringens disease, colibacillosis and Salmonella bacteriosis in livestock or poultry.