JP6946009B2 - Antibacterial active substance and fertilizer using it - Google Patents

Description

The present invention relates to antibacterial active substances and fertilizers made from shell fossils.

So far, when shells such as oysters, scallops, and scallops are fired at a high temperature of 1100 ° C. or higher, the calcium oxide content becomes 90% by weight or more, and the aqueous suspension exhibits a wide range of antibacterial activity and antiviral activity. It has been reported (Non-Patent Document 1, Patent Document 1).
The antibacterial activity and the like are due to the fact that the aqueous suspension has strong alkalinity.
Further, Patent Document 2 describes a bioactive substance obtained by heating a calcium-containing raw material, and Patent Document 3 describes a microbial inhibitor obtained by heating a pearl, and Patent Document 4 describes it. Discloses a bactericide obtained by firing oyster shells and a method for maintaining the freshness of food.
All of the techniques described in these publications are based on calcium oxide.
On the other hand, the present inventors have reached the present invention by using fossil shellfish, which will be described later, as a raw material.

Nagatoshi Oshima, "Application of Antibacterial Calcium" Monthly Food Chemicals, 14: 61-64 (1998)

Japanese Unexamined Patent Publication No. 2008-179555 Japanese Unexamined Patent Publication No. 1-117833 Japanese Unexamined Patent Publication No. 3-77802 Japanese Patent No. 3206602

An object of the present invention is to provide an antibacterial active substance which exhibits excellent antibacterial activity and is also useful as a fertilizer.

The antibacterial active substance according to the present invention is characterized by being composed of calcined shell fossil powder.
Here, shell fossils are those obtained from fossilized ore deposits of shellfish and the like.

Examples of shell fossils are those obtained from the Omma Formation, which is a stratum in the early quaternary era distributed from Kanazawa City in Ishikawa Prefecture to Oyabe City in Toyama Prefecture, and carbon dioxide in Toyama Prefecture. The main components are lime 80-85% and silicic acid 6-9%, and it contains trace components such as phosphate, potassium, magnesium, iron, manganese, boron, and molybdenum, and is a raw material for fertilizers and feeds called shell fossils. There is a deposit (Western Toyama Prefecture Shell Fossil Deposit).
The shellfish that make up the deposit are mainly bivalves and scallops, which are mixed with ancient organism remains such as foraminifera and brachiopods, fine sand and fine-grained gravel of Paleozoic rocks, and then uplifted due to crustal movements and then rotted.・ It is fossilized.
This shell fossil deposit extends to western Toyama Prefecture, Takaoka City, Fukuoka Town, Nishitonami District (currently Takaoka City) and Oyabe City. It is in the immediate vicinity of.
The strike line of the deposit is parallel to the Oyabe River channel, National Highway No. 8, and Ainokaze Toyama Railway (former JR Hokuriku Line).
The deposit runs vertically from the western corner of Toyama Prefecture to the southeastern line of the mountain ridge area on the border of Ishikawa Prefecture for a length of 12 km (for shell fossils produced in Toyama Prefecture, edited by the Toyama Prefectural Shell Fossil Fertilizer Association, November 1978, More quoted).
When shell fossils obtained from such deposits are used as raw materials, the deposits contain 6 to 9% by mass of silicic acid, and the calcined shell fossils contain 10 to 30% by mass of silicic acid, so that they are also useful as fertilizers. ..
Such fertilizers show antibacterial activity against many fungi, but are particularly useful in agriculture in that they show antibacterial activity against phytopathogenic fungi.

The antibacterial active substance according to the present invention exhibits antibacterial activity against bacteria including phytopathogenic fungi, actinomycetes, yeast, filamentous fungi and the like, as will be described later.

The test results of the calcination temperature and antibacterial activity of the raw powder of shellfish are shown. The effects of calcination time and calcination temperature on the antibacterial activity of scallop fossil powder are shown. FTIR Spectra of CaCO ₃ , CaO and calcined scallop shell powder is shown. FTIR Spectra of calcined shell fossil is shown. The results of potato cultivation are shown.

<Preparation method for shell fossil powder and baked products>
Shell fossils mined from the western shell fossil deposit in Toyama Prefecture are brought into Asahi Kako Co., Ltd. (Oyabe City, Toyama Prefecture), roughly crushed with a crusher (Hardware Brown Horizontal Crusher No. 1025-HB, manufactured by Yoshida Seisakusho), and then coarsely crushed. A stainless steel screen (ladder hole 0.5 mm) was attached to an ultracentrifugal crusher (ZM200, Lecce, Germany), and pulverization and sieving were performed.
Subsequently, a powder of 0.5 mm or less (hereinafter, this fossil shell powder is referred to as shell fossil powder) that has passed through the screen is fired in an electric furnace (FO810, manufactured by Yamato Chemical Co., Ltd., temperature control system ± 2 ° C. ( It was fired at 1150 ° C.).
<Measurement of particle size distribution of raw shell powder and calcined product>
In order to investigate the particle size distribution of the raw shell fossil powder and after firing, a 72-mesh (sieve opening 0.212 mm) sieve was used, and about 300 g of the raw shell fossil powder (particle size 0.212 mm or less) passed through the sieve was prepared. bottom.
The calcined product was prepared by calcining the obtained raw shell powder in an electric furnace (manufactured by FO810 Yamato Chemical Co., Ltd.) at 1100 ° C. for 1 hour.
<Measurement method>
Using each sieve (opening 0.212 mm and 0.106 mm), the weights of the sample remaining on the sieve and the sample passed through were measured by an electronic balance (manufactured by BA6100 Sartorius Japan), and the respective ratios were calculated.
<Analysis result>
The analysis results are shown in Table 1.
As a result, it was confirmed that a powdered calcined product could be obtained by using the powdered shell fossil.

Table 2 shows the results of component analysis of the raw shell powder and the calcined powder.
In the table, shell fossil bulk powder (1) and (2) show the component analysis results after the powdered bulk powder is dried, and shell fossil firing shows the analysis results of the powder fired at 1100 ° C. for 1 hour. ..
The analysis is based on the fertilizer analysis method (National Institute for Agro-Environmental Sciences, Ministry of Agriculture, Forestry and Fisheries), and the unit is mass%.
The deposit itself contains 6 to 9% by weight of silicic acid, including water, but the raw material obtained from the deposit was crushed and sieved, and as a result of analysis in a dry state, the total amount of silicic acid was obtained. It contained 10 to 30% and also contained other fertilizer components.
Shell fossils contain 0.05-0.3% phosphoric acid, 0.1-2.0% kari, 10-30% silicic acid (silicic acid), and 50-70% lime after firing. There is.

実験方法：肥料分析法（農林水産省農業環境技術研究法）１９９２年版記載の原子吸光測定法（４．５．１２）に従い、原子吸光機Ｚ−２３００、日立製作所製を使い、実施した。
＜ＦＴＩＲ（フーリエ変換赤外線吸収スペクトル）分析＞
フーリエ変換赤外分光光度計（ＦＴＩＲ）は、試料に赤外光を照射し、透過または反射した光量を測定して、分子の構造や官能基の情報をスペクトルから得て、物質定性・同定に関する情報を得るために用いられる。
貝化石を８００℃以上で焼成すると、主要成分のＣａＣＯ_３はＣａＯとＣＯ_２に分解される。
従って、ＣａＯに基づく赤外吸収スペクトルが、８００℃以上の焼成物には検出されるはずと考えられる。
先ずは、試薬のＣａＣＯ_３（純度９８％、和光純薬工業株式会社製）、ＣａＯ（純度９８％、和光純薬工業株式会社製）と焼成ホタテ貝殻パウダー（株式会社エイワンィー製）を使いＦＴＩＲ吸収スペクトル（ＦＴＩＲ：Ｓｐｅｃｔｒｍ１００，パーキンエルマージャパン製）を測定し、図３の結果を得た。
図３のように、焼成ホタテの３６４０ｃｍ^−１とＣａＯの３６４２ｃｍ^−１付近の鋭い吸収はＣａＯに基づく特徴的は吸収帯と考えられた。
続いて、貝化石原末を、焼成温度６００℃、７００℃、８００℃、９００℃、１０００℃と１１００℃の６段階で各１時間焼成した。
得られた各焼成物のＦＴＩＲ吸収スペクトルを図４に示した。
９００℃・１時間処理で、３６００ｃｍ^−１付近を鋭いピークはＣａＯ生成に基づくシグナルが認められるが、１０００℃と１１００℃では、３６００ｃｍ^−１付近の吸収は弱くなった。
これらの知見より、８００℃・１時間以上の焼成処理により、ＣａＯが生成され、特に９００℃・１時間処理が有効と見られた。
＜貝化石粉状原末の焼成温度と抗菌活性＞
貝化石の原末を、７００℃、８００℃、９００℃と１０００℃で、各１時間焼成した後、得られた焼成粉末を、被検菌を重層した寒天平板上に置床して培養し、抗菌活性に基づく阻止帯形成を観察した。
表４と図１に示す如く、７００℃・１時間の焼成物は抗菌活性を全く示さないが、８００℃以上で１時間焼成した生成物は全ての被検定菌に抗菌活性を示した。

上記実験に使用した培地組成は以下の通り。
ＹＭ培地：粉末酵母エキス（極東製薬）０．３％，Ｍａｌｔ ｅｘｔｒａｃｔ(Difco laboratories)０．３％，極東ペプトン（極東製薬）０．５％，Ｇｌｕｃｏｓｅ（和光純薬、特級）１．０％，硫酸マグネシウム七水和物（和光純薬、特級）０．１％、粉末寒天（和光純薬）１．５％。
ＧＢ培地：粉末酵母エキス（極東製薬）０．１％，ｂｅｅｆ ｅｘｔｒａｃｔ(Difco laboratories)，Ｎ−Ｚ−ａｍｉｎｅ（和光純薬）０．２％，マルトース水和物（和光純薬）１％， 粉末寒天（和光純薬）、ｐＨ７．３
ＬＢＧ培地：０．５％，Ｔｒｙｐｔｏｎｅ(Difco laboratories)１．０％，塩化ナトリウム（和光純薬、特級）１．０％，ｇｌｕｃｏｓｅ（和光純薬、特級）０．２％，粉末寒天（和光純薬）１．５％ <Effect of firing temperature on CaO formation from shell fossils>
The raw mussel powder was calcined at five stages of firing temperatures of 700 ° C., 800 ° C., 900 ° C., 1000 ° C. and 1100 ° C. for 1 hour each.
The calcium oxide (CaO) content of each of the obtained calcined products was measured.
As shown in Table 3, it was clarified that the CaO content was increased by the firing treatment at 800 ° C. or higher.
It was also confirmed that the pH of the 5% aqueous suspension supernatant of each powder did not change between the raw powder and the treatment at 700 ° C. for 1 hour, but at 800 ° C. or higher, the pH value increased as the calcination temperature increased. ..
For this reason, the firing temperature is preferably 800 ° C. or higher.

Experimental method: According to the atomic absorption measurement method (4.5.12) described in the 1992 edition of the fertilizer analysis method (Ministry of Agriculture, Forestry and Fisheries, National Institute for Agro-Environmental Sciences), the atomic absorption spectroscope Z-2300 and Hitachi, Ltd. were used.
<FTIR (Fourier Transform Infrared Absorption Spectrum) Analysis>
The Fourier transform infrared spectrophotometer (FTIR) irradiates a sample with infrared light, measures the amount of transmitted or reflected light, obtains information on the structure and functional groups of the molecule from the spectrum, and relates to material qualitative analysis and identification. Used to obtain information.
When the shell fossil is calcined at 800 ° C. or higher, the main component CaCO ₃ is decomposed into CaO and CO _2.
Therefore, it is considered that the infrared absorption spectrum based on CaO should be detected in the fired product at 800 ° C. or higher.
First, FTIR absorption using the reagents CaCO ₃ (purity 98%, manufactured by Wako Pure Chemical Industries, Ltd.), CaO (purity 98%, manufactured by Wako Pure Chemical Industries, Ltd.) and fired scallop shell powder (manufactured by Wako Pure Chemical Industries, Ltd.) The spectrum (FTIR: Spectorm100, manufactured by PerkinElmer Japan) was measured, and the result shown in FIG. 3 was obtained.
As shown in FIG. 3, sharp absorption 3642cm the vicinity ^-1 3640Cm ^-1 CaO in baked scallops were considered characteristic absorption band based on CaO.
Subsequently, the raw shell powder was calcined at 6 steps of firing temperatures of 600 ° C., 700 ° C., 800 ° C., 900 ° C., 1000 ° C. and 1100 ° C. for 1 hour each.
The FTIR absorption spectrum of each of the obtained fired products is shown in FIG.
In the treatment at 900 ° C. for 1 hour, a signal based on CaO formation was observed in the sharp peak around ^{3600 cm -1} , but the absorption around ^{3600 cm -1 was weakened at 1000 ° C. and 1100 ° C.}
From these findings, it was considered that CaO was produced by the baking treatment at 800 ° C. for 1 hour or more, and that the treatment at 900 ° C. for 1 hour was particularly effective.
<Baking temperature and antibacterial activity of fossil shell powder powder>
The raw powder of the fossil shell was fired at 700 ° C., 800 ° C., 900 ° C. and 1000 ° C. for 1 hour each, and then the obtained calcined powder was placed on an agar plate on which the test bacteria were layered and cultured. Inhibition zone formation based on antibacterial activity was observed.
As shown in Table 4 and FIG. 1, the product calcined at 700 ° C. for 1 hour showed no antibacterial activity, but the product calcined at 800 ° C. or higher for 1 hour showed antibacterial activity against all the test bacteria.

The medium composition used in the above experiment is as follows.
YM medium: powdered yeast extract (Far East Pharmaceutical) 0.3%, Malt extract (Difco laboratories) 0.3%, Far East Peptone (Far East Pharmaceutical) 0.5%, Glucose (Wako Pure Chemical Industries, Ltd.) 1.0%, Magnesium sulfate heptahydrate (Wako Pure Chemical Industries, Ltd., special grade) 0.1%, powdered agar (Wako Pure Chemical Industries, Ltd.) 1.5%.
GB medium: powdered yeast extract (Far East Pharmaceutical) 0.1%, beef extract (Difco laboratories), NZ-amine (Wako Pure Chemical Industries) 0.2%, maltose hydrate (Wako Pure Chemical Industries) 1%, powder Agar (Wako Pure Chemical Industries, Ltd.), pH 7.3
LBG medium: 0.5%, Tryptone (Difco laboratories) 1.0%, Sodium chloride (Wako Pure Chemical Industries, Ltd., special grade) 1.0%, glucose (Wako Pure Chemical Industries, Ltd., special grade) 0.2%, Powdered agar (Wako Pure Chemical Industries, Ltd.) Medicine) 1.5%

<Minimum blocking concentration of shellfish chemicals and shellfish fossil powder baked at 1100 ° C for 1 hour>
Using YM agar medium, the minimum blocking concentration (%) of the shell fossil raw powder water suspension and the shell fossil water suspension baked at 1100 ° C. for 1 hour was examined according to the conventional method of the agar plate dilution method.
As shown in Table 5, in the raw mussel fossil powder, no antibacterial activity was observed in all the test bacteria even when the maximum concentration of 0.5% was added, but in the calcined shell fossil, the growth of all the test bacteria was blocked.
In particular, the fact that it exhibits antibacterial activity against the pathogens of Lanxangia tsa and carrot root rot, which are important in agriculture, suggests the usefulness of fired shellfish fossils in agriculture.

<Effects of shell fossil firing temperature and firing time on antibacterial activity>
For the purpose of investigating the relationship between the firing temperature and firing time of shell fossils, prepared calcined products at 800 ° C and 1100 ° C for 30 minutes, 45 minutes and 1 hour, and agar plates using Escherichia coli NIHJ JC-2 as the test bacterium. The antibacterial activity was examined by the method.
As shown in Table 6 and FIG. 2, the antibacterial activity of the calcined product at a firing temperature of 800 ° C. for 30 minutes and the calcined product at 1100 ° C. for 60 minutes is weaker than that of other firing condition products. looks like.
From this, it can be said that the firing temperature is preferably 800 ° C. for 30 minutes or more and 1100 ° C. for 60 minutes or less.

<Effect of calcined shellfish fossil powder coating on autumn potato cultivation>
It has been clarified that the calcined shell fossil powder exhibits a wide range of antibacterial activities such as pathogens of potato scab disease, gram-positive bacteria, gram-negative bacteria, yeast, and filamentous fungi.
Therefore, using a field of continuous cropping disorder due to spring potato soybean disease (Akakura district, Oyabe city, spring potato variety: Baron), half-cut potatoes of autumn potatoes (Digima, Nishiyutaka) were fired at 1100 ° C for 1 hour. Fine powder of potato fossil was coated.
On August 31, 2016, various potatoes were planted with the cut side facing down.
Cultivation was carried out by customary law.
The harvest was carried out on December 24, 2016.
As shown in Table 7, the germination rate of the untreated plot was 50% for both varieties of Digima and Nishiyutaka, while the calcined shell fossil-treated plot coated with calcined shell fossil powder was superior at 83.3%. It became expensive.
In addition, the number of tubers per plant and the weight of tubers were also significantly higher in the calcined shell fossil powder treatment group.
FIG. 5 shows a photograph of the harvested autumn potatoes.
No outbreak of severe lanxangia tsa. was observed in spring potatoes.

Claims (1)

It has a step of pulverizing the shell fossil mined from the western shell fossil deposit in Toyama prefecture with a crusher to obtain the shell fossil powder, and a step of drying the shell fossil powder. The powder contains 10 to 30% fossil acid, 0.05 to 0.3% phosphoric acid, and 0.1 to 2.0% Kari.
It has a step of firing the shell fossil powder in a temperature range of 800 ° C. to 1100 ° C.
A method for producing an antibacterial active substance, which contains 50 to 70% of lime after firing and has antibacterial activity.

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