KR102702960B1 - Antibiotic compounds for feed disinfection - Google Patents

Abstract

The purpose of the present invention is to provide an antimicrobial composition for disinfecting livestock feed, which has improved antimicrobial activity by mixing two or more organic acids with a cationic surfactant in a specific ratio. The antimicrobial composition for disinfecting livestock feed comprises: 1 to 70 wt% of formic acid; 1 to 25 wt% of citric acid; 0.1 to 5 wt% of didecyl dimethyl ammonium chloride; and the balance of water. According to the present invention, since two or more organic acids are mixed with a cationic surfactant in a specific ratio, an antimicrobial composition for disinfecting livestock feed exhibits progressive pH reduction, excellent cellular penetration, and outstanding antimicrobial activity, and thus can resolve physiological side effects, usage inconveniences, and public health-related environmental sanitation problems.

Description

{Antibiotic compounds for feed disinfection}

The present invention relates to an antibacterial composition for disinfecting livestock feed.

Livestock need energy to grow and move, and most of this is obtained through feed. However, if the feed is contaminated with microorganisms or the raw materials for producing the feed are contaminated, the livestock will inevitably be exposed to pathogenic microorganisms and will be susceptible to diseases, which will in turn lead to decreased productivity and the production of unstable livestock products. Although continuous efforts are being made to prevent such contamination of feed, contamination by Salmonella , Escherichia coli , and Staphylococcus aureus , which cause enteritis and diarrhea, is still a problem, and this has a significant negative impact on feed quality and livestock productivity.

Organic acids are components for normal metabolism and also play an important role in animal nutrition as natural products, and have the effect of promoting animal growth by lowering acidity in the digestive tract, increasing pepsin activity, suppressing bacterial growth, and increasing intestinal nutrient digestibility. In addition, they have antibacterial activity, so they control intestinal bacterial flora and play an important role in animal health, including the intestinal immune system. Specifically, organic acids have the effect of lowering pH (acidifier) with cations and the effect of interfering with protein production metabolism with anions, so they are widely used to suppress pathogens such as salmonella that contaminate feed raw materials or compound feed.

Among various organic acids, acetic acid has a sterilizing ability, and it prevents spoilage by killing bacteria that cause food poisoning such as E. coli and Staphylococcus aureus, and lactic acid has the effect of preventing the growth of bacteria and suppressing abnormal fermentation in fermented foods due to its own acidity. In addition, formic acid has been reported to have an MIC (Minimum inhibitory concentration) of 0.6 to 1.1 mmol/ℓ in the air for microorganisms, and it is possible to suppress the growth of microorganisms even in relatively small amounts, and it has been reported to be effective against various bacteria such as Salmonella, E. coli, and Enterobacter . It has been reported that citric acid can inhibit the growth or sterilize Staphylococcus aureus and Psedomonas aeruginosa with a use of about 0.5%.

In this way, various organic acids have pathogen suppression effects, but there is a problem that antibacterial agents containing only one type of organic acid have insufficient antibacterial activity and that the organic acid must be used at high concentrations. In addition, antibacterial agents using formaldehyde have side effects, are inconvenient to use, and have public health environmental hygiene issues, making it difficult to use them in the future. Therefore, the development of a safe and environmentally friendly antibacterial composition that resolves the above problems is required.

Republic of Korea Patent No. 10-2221983 (announced on March 2, 2021)

The problem to be solved by the present invention is to provide an antibacterial composition for disinfecting livestock feed having improved antibacterial activity by mixing two or more organic acids with a cationic surfactant at a specific ratio.

Another problem that the present invention seeks to solve is to provide an antibacterial composition for disinfecting livestock feed that can solve problems such as inconvenience in use, public health environmental hygiene problems, and decreased productivity of livestock.

The present invention, in one embodiment, provides an antibacterial composition for disinfecting livestock feed, comprising an organic acid and a cationic surfactant, wherein the organic acid is at least one selected from the group consisting of formic acid, propionic acid, lactic acid, benzoic acid, sorbic acid, fumaric acid, and acetic acid.

Additionally, the organic acid can be formic acid and citric acid.

In addition, it may contain 1 to 70 wt% of formic acid, 1 to 25 wt% of citric acid, 0.1 to 5 wt% of cationic surfactant, and the remainder of water.

Additionally, the cationic surfactant may be didecyl dimethyl ammonium chloride.

According to the present invention, by mixing two or more organic acids with a cationic surfactant at a specific ratio, an antimicrobial composition for disinfecting livestock feed can be provided, which has excellent antimicrobial activity with synergistic pH lowering and excellent cell penetration ability, thereby solving physiological side effects, inconvenience in use, and public health environmental hygiene problems.

In addition, by applying the antibacterial composition for livestock feed disinfection to livestock feed raw materials, compound feed, and feed production processes, high-quality feed can be supplied to livestock, and stability can be secured, thereby increasing livestock productivity.

Figure 1 shows the results of analyzing the antibacterial activity of Example 3 against Sg, Se, St, Escherichia coli, and Staphylococcus aureus in a test tube.

Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings so that those skilled in the art can easily practice the present invention. However, the present invention may be implemented in various different embodiments and is not limited to the embodiments described herein. Like parts are designated by the same reference numerals throughout the specification.

The inventors of the present invention, while studying organic acids having antibacterial active ingredients, discovered that a synergistic effect was exhibited when specific organic acids were mixed and used at a specific ratio, and thus completed the present invention.

The present invention relates to an antibacterial composition for disinfecting livestock feed that can exhibit an antibacterial synergy effect through a combination of organic acids at a specific ratio.

The present invention provides, in one embodiment, an antibacterial composition for disinfecting livestock feed, comprising an organic acid and a cationic surfactant.

It is desirable to use organic acids that have high dissociation, excellent acidity-lowering power, easy intracellular penetration, and excellent antibacterial properties.

Specifically, the organic acid may be at least one selected from the group consisting of formic acid, propionic acid, lactic acid, benzoic acid, sorbic acid, fumaric acid, and acetic acid, and more specifically, formic acid and citric acid may be used in combination.

Formic acid has the simplest structure among aliphatic organic acids, has a high degree of dissociation in aqueous solution, and has excellent pH lowering ability. It also has the advantage of easy penetration into cells due to its small molecular structure.

Cationic surfactants have antibacterial properties of their own, while also playing a role in stabilizing antibacterial compositions.

The cationic surfactant may be, but is not limited to, didecyl dimethyl ammonium chloride (DDAC).

DDAC can dissolve cell membranes composed of double phospholipids, has its own antibacterial properties, and helps organic acids easily penetrate into cells, thereby increasing antibacterial activity, as well as stabilizing the properties of the composition and protecting it from the external environment. In addition, it is very useful because it does not irritate the body and the human body, is not corrosive to equipment, and does not decrease antibacterial activity even when diluted in hard water.

An antibacterial composition for disinfecting livestock feed may contain 1 to 70 wt% of formic acid, 1 to 25 wt% of citric acid, 0.1 to 5 wt% of a cationic surfactant, and the remainder of water.

If the content of formic acid is less than 1 wt%, the antibacterial activity is reduced and a synergistic effect cannot be achieved when combined with citric acid. If the content of formic acid exceeds 70 wt%, not only is a synergistic effect not achieved due to the excessive content compared to citric acid, but there is also a problem of reduced economic feasibility.

If the content of citric acid is less than 1 wt%, it does not exhibit a synergistic effect when combined with formic acid, and if the content of citric acid exceeds 70 wt%, formic acid dissociates and the antibacterial effect is reduced, and the excessive content compared to formic acid does not exhibit a synergistic effect.

If the content of the cationic surfactant is less than 0.1 wt%, the stabilizing effect of the antibacterial composition cannot be exerted, and if it exceeds 5 wt%, the antibacterial effect may be reduced, so the above range is preferable.

Hereinafter, the present invention will be described in more detail using examples. It will be apparent to those skilled in the art that these examples are only intended to explain the present invention more specifically and that the scope of the present invention is not limited by these examples.

Search for optimal organic acid combinations

1. Evaluation of antibacterial activity of organic acids

1㎖ of pre-cultured Salmonella enteritidis was added to 9㎖ of raw material saline solution, and the initial bacterial count was measured. Then, each raw material was added in the amount (0.01 g:0.1%), and the number of surviving bacteria after 3 hours was measured to compare the antibacterial activity. The initial bacterial count was measured in plates of 4, 5, and 6, and the bacterial count after 3 hours was measured in plates of 2, 3, and 4, and the results are shown in Table 1.

sample Count of bacteria (cfu/g) after 3 hours Control group (initial bacterial count: 6.1×10 ⁷ ) 67,000,000 Formic acid 1.18 g 120,000 1 g of propionic acid 7,000,000 1.25 g of lactic acid 5,600,000 1.09 g of citric acid 5,500,000 1 g of benzoic acid 7,200,000 1 g of sorbic acid 5,500,000 1 g of fumaric acid 2,700,000 1 g of sodium propionate 8,600,000 Calcium sorbate 1 g 8,500,000 1 g of acetic acid 6,400,000

Referring to Table 1, it was confirmed that all the tested organic acids had antibacterial activity. Based on the same amount, formic acid had the best antibacterial activity, followed by fumaric acid, citric acid, sorbic acid, lactic acid, acetic acid, propionic acid, benzoic acid, potassium sorbate, and sodium propionate. Among these, potassium sorbate and sodium propionate, which are in salt form, had the worst antibacterial activity, and formic acid was confirmed to have an effect that was about 10 times more effective than other organic acids. Fumaric acid showed the second highest antibacterial activity, but fumaric acid has low solubility in water and is not suitable for use with formic acid. Citric acid showed the third highest antibacterial activity, and since citric acid has a similar first-order dissociation degree to formic acid and is suitable for maintaining formic acid in its original form, it facilitates intracellular penetration, so it is expected that it will enhance the antibacterial activity of organic acids and exhibit much stronger antibacterial activity in combination with formic acid.

2. Comparison of antibacterial activity of formaldehyde and cationic surfactant (DDAC)

The initial bacterial count was measured by adding 1 ml of pre-cultured Salmonella Enteritidis to 9 ml of saline solution, and then each sample (0.01 g:0.1%) was added, and the surviving count was measured after 3 hours, and the results are shown in Table 2.

division Usage bacteria Initial count
(cfu/g) Count after 3 hours
(cfu/g) Sterilization rate (%) Mixture of DDAC 12.5g and fruit 87.5g (Sample 1) S. enteritidis 4.6×10 ⁶ 0 100 A mixture of 37 g of formaldehyde and 63 g of fruit (control) S. enteritidis 3.3×10 0 100

Referring to Table 3, it can be confirmed that DDAC has excellent antibacterial properties according to the measurement results and is a suitable substance to replace formaldehyde. Therefore, the present invention may preferably include a cationic surfactant in addition to formic acid and citric acid as antibacterial active ingredients.

<Example>

Example 1

An antibacterial composition for disinfecting livestock feed was prepared by mixing 15.0 g of formic acid, 5.0 g of citric acid, 1.0 g of DDAC, and 79 ml of water.

Example 2

An antibacterial composition for disinfecting livestock feed was prepared by mixing 30.0 g of formic acid, 15.0 g of propionic acid, 1.0 g of DDAC, and 54 ml of water.

Example 3

An antibacterial composition for disinfecting livestock feed was prepared by mixing 30.0 g of formic acid, 5.0 g of citric acid, 1.0 g of DDAC, and 64 ml of water.

<Comparative Example>

Comparative Example 1

An antibacterial composition was prepared by mixing 24.0 g of formaldehyde, 7.0 g of formic acid, 16 g of propionic acid, and 53 ml of water.

Comparative Example 2

An antibacterial composition was prepared by mixing 24.0 g of formaldehyde, 19.0 g of formic acid, 10.7 g of propionic acid, and 46.3 ml of water.

Comparative Example 3

An antibacterial composition was prepared by mixing 30.0 g of formaldehyde, 15.0 g of formic acid, and 55.0 ml of water.

<Example of an exam>

Test Example 1: Antibacterial activity evaluation

The antibacterial activity of the antibacterial compositions manufactured according to Examples 1 and 2 and Comparative Examples 1 to 3 was evaluated.

In order to comparatively evaluate the antibacterial activity, 1 ml of pre-cultured Salmonella bacteria (Salmonella enteritidis (hereinafter Se), Salmonella gallinarium (hereinafter Sg), Salmonella typhimurium (hereinafter St)) was added to 9 ml of saline solution, and the initial bacterial count was measured. Then, the antibacterial compositions manufactured according to Example 1, Example 2, and Comparative Examples 1 to 3 were added in amounts (0.01 g:0.1%), respectively, and the number of survivors was measured after 3 hours.

The initial bacterial counts were measured in plates of the 4th, 5th, and 6th powers, and the bacterial counts after 3 hours were measured in plates of the 1st and 2nd powers, and the results are shown in Table 3 below.

division Usage bacteria Initial count (cfu/g) Count of bacteria (cfu/g) after 3 hours Example 1 Sg 7.3×10 ⁷ 1×10 ² Se 1.2×10 ⁸ 0 St 8.6×10 ⁷ 0 Example 2 Sg 7.3×10 ⁷ 0 Se 1.2×10 ⁸ 0 St 8.6×10 ⁷ 1.0×10 ³ Comparative Example 1 Sg 7.3×10 ⁷ 0 Se 1.2×10 ⁸ Countless St 8.6×10 ⁷ Countless Comparative Example 2 Sg 7.3×10 ⁷ 0 Se 1.2×10 ⁸ Countless St 8.6×10 ⁷ Countless Comparative Example 3 Sg 7.3×10 ⁷ 0 Se 1.2×10 ⁸ 0 St 8.6×10 ⁷

Referring to Table 3, it can be confirmed that the antibacterial activity of the antibacterial composition of Comparative Example 3 containing the highest amount of formaldehyde was the highest, but exhibited antibacterial activity similar to that of the antibacterial compositions manufactured according to Examples 1 and 2 that did not use formaldehyde. That is, the antibacterial composition manufactured according to the present invention can replace the conventional antibacterial composition containing formaldehyde, thereby resolving the side effects of formaldehyde, inconvenience in use, and public health environmental hygiene issues.

Test Example 2: Analysis of antibacterial activity according to mixing conditions

In Test Example 1, it was confirmed that the antibacterial composition of the present invention can replace an antibacterial composition containing formaldehyde as an antibacterial active ingredient, and the antibacterial power according to the mixing of organic acids was analyzed.

The test conditions were the same as in Test Example 1, and the antibacterial activity of Examples 1, 2, 3 and Comparative Example 3 was analyzed twice by changing the initial bacterial count. Tables 4 and 5 below show the results.

division
Initial count (cfu/g) Count of bacteria (cfu/g) after 3 hours Example 1 Sg 6.3E+06 90 Se 2.3E+07 110 St 3.7E+07 100 Example 2 Sg 6.3E+06 150 Se 2.3E+07 0 St 3.7E+07 540 Example 3 Sg 6.3E+06 33 Se 2.3E+07 20 St 3.7E+07 63 Comparative Example 3 Sg 6.3E+06 93 Se 2.3E+07 58 St 3.7E+07 190

division
Initial count (cfu/g) Count of bacteria (cfu/g) after 3 hours Example 1 Sg 1.6E+05 10 Se 1.2E+06 53 St 3.5E+06 0 Example 2 Sg 1.6E+05 0 Se 1.2E+06 0 St 3.5E+06 17 Example 3 Sg 1.6E+05 0 Se 1.2E+06 0 St 3.5E+06 4 Comparative Example 3 Sg 1.6E+05 10 Se 1.2E+06 20 St 3.5E+06 0

Referring to Tables 4 and 5, the antibacterial composition according to Example 3 exhibited the highest antibacterial activity, and when only the examples are compared with each other, first of all, the antibacterial activity of the antibacterial composition containing formic acid and citric acid, Example 2, was superior to the antibacterial activity of the antibacterial composition containing formic acid and propionic acid, depending on the type of organic acid blended. This was despite the fact that the propionic acid content of Example 2 was much higher than the citric acid content of Example 3, and it was found that citric acid was more useful in blending with formic acid than propionic acid. When the same organic acids were blended and used, the antibacterial activity of the antibacterial composition of Example 3 having a high formic acid content was much superior to the antibacterial activity of the antibacterial composition of Example 1 having a low formic acid content, confirming that the higher the content of formic acid, the better the antibacterial activity.

Test Example 3: Antibacterial activity evaluation according to harmful bacteria

The antibacterial activity of the antibacterial composition of Example 3 against various pathogens, such as Sg, Se, St, Escherichia coli, and Staphylococcus aureus, was evaluated in a test tube, and the results are shown in Table 6 below.

In addition, livestock feed and feed ingredients were contaminated with pathogens and treated with Example 3 to evaluate the antibacterial activity, which is shown in Table 7 and Fig. 1.

division Bacterial count (cfu/g) Initial count 0 hours after addition 3 hours after addition Sg 2.8×10 ⁵ 1.3×10 ² Below detection limit Se 3.4×10 ⁵ 1.1×10 ² Below detection limit St 3.6×10 ⁵ 3.8×10 ² Below detection limit Escherichia coli 4.2×10 ⁵ Below detection limit Below detection limit Staphylococcus aureus 2.9×10 ⁵ Below detection limit Below detection limit

Bacterial count (cfu/g)
Initial mixed total bacteria Total germs after 3 hours Total germs after 24 hours Total bacteria after enrichment Laying hen feed 3.3×10 ⁴ 7.0×10 ⁴ 4.6×10 ⁴ Below detection limit Sujigak 3.3×10 ⁴ 3.3×10 ⁴ Below detection limit Below detection limit Fish meal 3.3×10 ⁴ Below detection limit Below detection limit Below detection limit Umobun 3.3×10 ⁴ Below detection limit Below detection limit Below detection limit Meat and bone meal 3.3×10 ⁴ Below detection limit Below detection limit Below detection limit

Referring to Tables 6 and 7 and Fig. 1, it was confirmed that the antimicrobial composition of Example 3 not only exhibited excellent in vitro efficacy against each pathogen, but also exhibited excellent efficacy against animal protein raw materials such as laying hen feed, slurry, fish meal, feather meal, and meat and bone meal, which are actually applicable livestock feed and feed raw materials.

Test Example 4: Efficacy Verification for Protein Meat Powder

The antibacterial composition of Example 3 was applied to a factory producing 20 tons of chicken meal, an animal protein used as a livestock feed ingredient, per day, and its efficacy was verified.

The verification method is to add 0.2% of the antimicrobial composition manufactured according to Example 3, add 90 ml of sterilized saline solution, and mix well by adding 10 g each of meat powder samples A, B, and C to a 300 ml Erlenmeyer flask. After leaving it for approximately 10 minutes, 1 ml each is collected from the upper layer of the mixture and tested on a standard agar medium (Plate count agar, PCA) and MCA(Mac Conkey Agar) Spread on a plate, dry, and culture in an incubator maintained at 37℃ for 2 days, then observe the number of bacterial colonies. In addition, 1㎖ of the supernatant was collected and entered into a Salmonella confirmation kit (Kit) to confirm the presence or absence of Salmonella, which is shown in Table 8 below.

division Colony count measurement
result Salmonella
existence and nonexistence Mac ConKey Agar PCA A Below detection limit Below detection limit No contaminants voice B Below detection limit Below detection limit No contaminants voice C Below detection limit Below detection limit No contaminants voice

Referring to Table 8, it was found that Salmonella and contaminants were not detected in any of the chicken meal samples A, B, and C, and all were also confirmed negative by the Salmonella confirmation kit. As a result, it was confirmed that the antibacterial composition according to the present invention has excellent antibacterial activity against various pathogens, and in particular, it can replace formaldehyde, thereby providing an eco-friendly antibacterial composition that solves physiological side effects, inconvenience in use, and public health environmental hygiene problems. In addition, it was confirmed that it is very effective in the disinfection of various livestock feeds, feed raw materials, and feed production processes.

Claims (4)

Containing 1 to 70 wt% of formic acid, 1 to 25 wt% of citric acid, 0.1 to 5 wt% of didecyl dimethyl ammonium chloride, and the remainder of water.
Antibacterial composition for disinfecting livestock feed. delete delete delete