KR20140137529A - Cinnamon bark essential oil having anti-bacterial activity against causative bacteria of porcine respiratory disease complex and uses thereof - Google Patents

Abstract

The present invention relates to an anti-bacterial composition including cinnamon bark essential oil for inhibiting pathogenic bacteria of domestic animals which specifically inhibits Actinobacillus pleuropneumoniae, Pasteurella multocida A, and Pasteurella multocida D which are causative bacteria of porcine respiratory disease complex, and to livestock feed additives and livestock feed comprising the composition as an active ingredient. The anti-bacterial composition including cinnamon bark essential oil for inhibiting pathogenic bacteria of domestic animal can be used as an alternative antibiotic, thereby can be valuably used in industrial fields.

Description

[0001] The present invention relates to a cinnamon essential oil having antimicrobial activity against bacterial causative bacteria of a porcine respiratory tract, and to a use thereof.

The present invention relates to a cinnamon essential oil having antimicrobial activity against bacteria causing porcine respiratory infections and more particularly to an antimicrobial composition for inhibiting bacterial pathogenic bacteria containing cinnamon essential oil as an active ingredient, Actinobacillus < / RTI > ( Actinobacillus < RTI ID = 0.0 > pleuropneumoniae , < / RTI > Pasteurella The present invention also relates to an antimicrobial composition for inhibiting a multocida A and a pastelolactone D, and a livestock feed additive and a livestock feed containing the composition as an active ingredient.

PRDC (porcine respiratory disease complex) is a disease characterized by poor growth, poor feed efficiency, lethargy, anorexia, fever, cough, and respiratory distress, occurring mainly in stock and finishing pigs between 10 and 22 weeks of age. Porcine respiratory syncytial virus infection can be caused by a variety of diseases including porcine respiratory syndrome virus (PRRSV), swine influenza virus (SIV), pseudorabies virus, Mycoplasma hyopneumoniae , Actinobacillus pleurpneumoniae and Pasteurella < RTI ID = 0.0 > Multicida ), among them, Actinobacillus A. pleuropneumoniae and P. multocida are the primary infectious agents of porcine respiratory syncytial virus infection Secondary infections cause severe symptoms. Actinobacillus pneumoniae is a causative organism of acute and chronic pneumothorax. It is associated with fibrosing pleurisy and haemorrhagic necrotizing lesions. The respiratory infectious disease caused by this bacterium is often caused by a poor environment or a dense breeding condition, and pigs infected with this disease cause a delay in development including mortality and a decrease in feed efficiency, thereby causing a great economic loss to the pigs. This strain acts as a primary infectious organism in the respiratory system of pigs. It is said that 10 ⁴ to 10 ⁵ bacteria per animal are killed intravenously during artificial infection, and 10 ³ microorganisms are injected intravenously to take chronic infection . Pasteurla La Mortosida is a pathogen in the swine of the respiratory tract that induces respiratory disease. It is classified as A, B, D, E and F according to the serotype of the cavernous sinus. The toxicity is determined by the presence of the toxA gene, which encodes a 145 kDa toxin protein, and is predominantly expressed in type A or type D. Type A is known to cause bronchopneumonia in pigs, cattle, and chickens, hemorrhagic sepsis in cattle in type B, and atropic rhinitis (AR) in pigs.

It is not easy to know the severity of disease in swine farms because swine respiratory diseases are often infectious or chronic. Therefore, the possibility of swine respiratory disease is increasing due to the large scale of modern swine industry. Is more difficult. Currently, the use of antibiotics and antibiotics in livestock is being strengthened due to the safety issue of livestock products. In particular, regulations on the use of antibiotics added to compound feeds will be strengthened in the future. Avoiding the use of antibiotics. In addition, the abuse of antibiotics causes the development of antibiotic resistant pathogens.

Recently, researches on environmentally friendly antibiotic substitutes have been carried out in various fields. In particular, researches on probiotics using useful microorganisms showing direct antagonistic action against pathogenic microorganisms and materials showing antimicrobial activity from plant resources and other natural substances Has been developed and commercialized. Studies using antimicrobial agents, which exhibit antimicrobial activity against various pathogenic microorganisms present in plant resources, as antimicrobial materials have been actively studied in food, medicine, and biotechnology industries for a long time (Shin et al., 1997 , J. Food SCI. TECHNOL. 29: 808-816), herbicides of various plant sources have not only a direct antimicrobial activity but also various bioactive chemical compounds and a variety of vitamins, calcium and iron It has nutrients that have been applied to the prevention and treatment of diseases. Especially, spice plants such as ginseng, incense acacia, wormwood, chrysanthemum, iris, rhododendron, rosemary, rhododendron, cinnamon, cinnamon, juniper, citron and tangja have excellent physiological activity and are widely used.

Among them, cinnamon is the most common cinnamomum cassia Blume is a kind of bark in which the bark or juniper of a relative plant is removed to some extent. It is also referred to as a jade tree or a big tree. The mountainous region is mainly southern China, southern Japan, Vietnam, Cambodia, Thailand, And it is a plant resource that has been used for a long time as a spice such as confectionary, drink, and herb medicine. Cinnamon bark contains cinnamic aldehyde and cinnamic acid, which are the major components of tannin and essential oil. Cinnamon oil is an oily matter which is distilled from cinnamon. There is a lot of sweetness and sincerity. The pharmacological actions of cinnamon have been reported as improving bacteria inhibiting action, inhibition of white spore strain, actinic action, saliva and gastric secretion promoting action, and the active ingredient is volatile essential oil component of about 1%. Phellandrene ), Eugenal, Methyleugual, and the like.

Korean Patent Publication No. 2003-0031331 discloses 'antifungal and insecticidal composition containing cinnamon extract', Korean Patent No. 693077 discloses 'biological composition for prevention and treatment of porcine combined respiratory disease' , The cinnamon essential oil showing antimicrobial activity against the bacteria causing the porcine respiratory tract infection of the present invention and its use are not described.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to provide a method for the production of actinobacillus flucytosine pleuropneumoniae , < / RTI > Pasteurella multocida A and pasturella mushroom D, the present inventors completed the present invention by confirming that cinnamon essential oil exhibits antimicrobial activity strongly inhibiting the growth of the pathogenic bacterium.

In order to solve the above problems, the present invention provides an antimicrobial composition for inhibiting bacterial pathogenic bacteria containing cinnamon bark essential oil as an active ingredient.

The present invention also provides a method of preventing or treating pathogenic bacterial diseases caused by livestock, which comprises treating the above-mentioned antimicrobial composition for inhibiting livestock pathogenic bacteria.

In addition, the present invention provides a livestock feed additive and a livestock feed containing an antimicrobial composition for inhibiting bacterial pathogenic bacteria containing the cinnamon essential oil as an active ingredient.

In the present invention, cinnamon essential oil showed a strong antimicrobial activity against causative microorganisms causing porcine respiratory infections, confirming the possibility of a new antibiotic substitute. Therefore, the cinnamon essential oil of the present invention can be used as an alternative antibiotic for the prevention and treatment of diseases of livestock, and thus can be very useful industrially.

1 is of the well close to that contain cinnamon essential oil as a result of the agar-well diffusion method (agar well diffusion method) Measurement of antibacterial activity against pneumoniae in liquid Martino Bacillus flu cinnamon essential oil (A. Pleuropneumoniae) colonies Growth was suppressed. DMSO; Negative control, Ampicillin; Positive control.

In order to achieve the object of the present invention, the present invention provides an antimicrobial composition for inhibiting bacterial pathogenic bacteria containing cinnamon bark essential oil as an active ingredient.

By "antimicrobial" is meant the ability to reduce, prevent, inhibit, or eliminate microbial growth or survival at any concentration.

The term " comprising "of the present invention is not limited to inclusive, and does not exclude, for example, other additives, ingredients, exponents, or steps.

The cinnamon essential oil of the antimicrobial composition for inhibiting bacterial pathogenic bacteria of the present invention may be one extracted by a solvent extraction method, a supercritical carbon dioxide extraction method or a steam distillation method, but is not limited thereto.

The livestock pathogenic bacterium of the present invention is a bacterium that induces Porcine Respiratory Disease Complex, which is called Mannheimia haemolytica ), Haemopilus somnus , Pasteurella multocida , Mycoplasma hyopneumoniae or Actinobacillus < RTI ID = 0.0 > pleuropneumoniae ), preferably Actinobacillus < RTI ID = 0.0 > pleuropneumonia e), Pasteurella multocida A, or passthrough, but is not limited thereto.

The present invention also provides a method for preventing or treating pathogenic bacterial diseases caused by livestock, which comprises treating a livestock with an antimicrobial composition for controlling livestock bacterium containing cinnamon essential oil as an active ingredient.

In the method of the present invention, the step of treating the livestock with the antimicrobial composition for inhibiting livestock bacterium may be spraying or vaporizing in a livestock farm, but is not limited thereto.

In a method according to an embodiment of the present invention, the animal may be a cow, a chicken, a pig, a horse, a goat, a duck, a goose, a dog, a cat or a rabbit, and preferably a pig.

In the method of the present invention, the livestock pathogenic bacterium is a bacterium that induces Porcine Respiratory Disease Complex, which is Mannheimia haemolytica ), Haemopilus somnus , Pasteurella multocida), mycoplasma (Mycoplasma hyopneumoniae) or liquid Martino Bacillus flu in pneumoniae (Actinobacillus pleuropneumoniae) may be, and preferably liquid Martino Bacillus flu in pneumoniae (Actinobacillus pleuropneumonia e, Pasteurella multocida A, or Passthrough la water D, but not limited thereto.

The present invention also provides a livestock feed additive and a livestock feed containing as an active ingredient an antimicrobial composition for controlling livestock bacteria containing cinnamon essential oil.

The livestock feed additive of the present invention can be used as it is in its original form, or additionally, known carriers such as cereal grains and by-products thereof, stabilizers and the like can be added, and if necessary, citric acid, fumaric acid, adipic acid, lactic acid, Such as polyphosphoric acid, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid (phosphoric acid) Antibiotics, antimicrobial agents, and other additives may be added. The shape of the powder may be a proper state such as powder, granule, pellet, suspension, etc. When the feed additive is supplied, Or mixed with feed.

When preparing livestock feeds in the present invention, the additive may be selected from cereal crops such as ground or grinded wheat, oats, barley, corn and rice; Vegetable protein sources based on rapeseed, soybean and sunflower seeds; An animal protein source; molasses; And milk products, such as various milk powders and whey powders. After mixing with all the dry ingredients, the liquid ingredients and the ingredients that become liquid after heating can be added. The liquid component may optionally consist of lipids liquefied by heating, for example fats, for example vegetable fats, and / or carboxylic acids, for example fatty acids. After thorough mixing, a powder or particulate concentration is obtained depending on the degree of polishing of the component. In order to prevent separation during storage, water should preferably be added to the animal feed, which is subsequently treated with conventional pelletization, extrusion or extrusion processes. Any additional water can be removed by drying. If desired, the resulting particulate animal feed can be pulverized into smaller particle sizes.

In a livestock feed additive and a livestock feed according to an embodiment of the present invention, the livestock is preferably a cow, a chicken, a pig, a horse, a goat, a duck, a goose, a dog, a cat or a rabbit, But is not limited thereto.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  1. Antimicrobial activity measurement of vegetable essential oil

Agar diffusion assay is used for general antimicrobial activity measurement method. There are two methods of agar diffusion method: paper disc method and agar well method. The principle is that the material having antimicrobial activity diffuses into the agar and the size of the clear zone In the case of the paper disk method, there is a disadvantage that the antimicrobial substance may be condensed or volatilized on the disk. As a result of comparing antimicrobial activities by two methods of paper disk method and agarwel method in measuring the antimicrobial activity of vegetable essential oil component, it was concluded that the agarwel method is more suitable than the paper disk method, .

Antibacterial activity against pneumoniae (A. Pleuropneumoniae) to purchase and a total of 14 vegetable oil refining with liquid Martino Bacillus flu by the agar-well diffusion method as a primary measure. Actino Bacillus fluroneumoniae 100 占 퐇 of the culture medium of the strain (OD ₆₀₀ = 0.5 to 0.6) was uniformly plated on the surface of the TSB agar medium and wells were formed on the surface of the plate. 50 쨉 l of vegetable essential oil and indicator material were added to each well and cultured at 30 째 C for 24 hours. The diameters of growth inhibition rings inhibited the growth of the indicated strains around the agar well, and the magnitude of the antimicrobial activity was shown. 1 mg / ml of antibiotic ampicillin was used as a positive control. The results of the antimicrobial activity measurements are shown in Table 1.

Cinnamon bark and thyme essential oil showed the best antimicrobial activity in the other 14 vegetable essential oils, showing the same inhibitory growth inhibition size of 15 mm as the positive control. Eucalyptus and bay essential oil produced the second highest growth inhibition size of 10 mm. The remaining 10 vegetable essential oils have an antimicrobial activity of about 1 to 5 mm in inhibition of growth on Actinobacillus fluroxenone, but are not superior in antibacterial activity to positive control, cinnamon, thyme, eucalyptus and bay oil .

Example  2. Inhibitory concentration measurement of vegetable essential oil

A liquid Martino Bacillus flu pneumoniae in (A. Pleuropneumoniae) bacteria inhibitory concentrations (inhibitory concentration) in the fine cinnamon, thyme, bay essential oil of eucalyptus and the antimicrobial activity for on the basis of the results of experiments in oil refining raw liquid was measured. The vegetable essential oil was diluted 1 to 100000 ppm with 1/10 dilution of DMSO (dimethyl sulfoxide, St. Louis, Mo., USA) and the inhibitory concentration was measured by an agar well diffusion method. DMSO used for dilution was used as a negative control, and antibiotic ampicillin was used as a positive control. The results are shown in Table 2.

In the case of 100000ppm concentration, cinnamon essential oil showed the size of growth inhibition ring of 15 mm same as undiluted undiluted solution, and time essential oil showed growth inhibition ring of 10 mm smaller than undiluted solution. In case of eucalyptus and bay oil, And 6 mm and 3 mm in size, respectively. At the concentration of 10000ppm, cinnamon essential oil showed growth inhibition of 5 mm, which was about 1/3 less than the original solution. Time essential oil produced a growth inhibition ring of 2 mm, which was about 1/5 smaller than the undiluted solution. Eucalyptus and bay oil At the concentration of 10000 ppm, no inhibition of growth was observed. It was confirmed that no inhibitory effect on growth inhibition was observed in all of the four vegetable essential oils at a concentration of 1000 ppm or less, indicating no antimicrobial activity.

Example  3. Measurement of minimum inhibitory concentration of cinnamon essential oil

Actinobacillus pluropneumoniae of cinnamon essential oil, which showed excellent antimicrobial activity as a result of agar diffusion method and inhibition concentration measurement, A and the minimum inhibitory concentration (MIC) for the passthrough la Water Soda D were measured.

First, the cinnamon essential oil was diluted with DMSO and added to TSB and BHI liquid medium. The total volume was adjusted to 3 ml. Each strain was inoculated into the culture medium and cultured at 30 ° C for 24 hours. Minimal inhibition concentration analysis was performed by modifying the broth dilution method. DMSO was used as a negative control, and ampicillin was used as a positive control. Table 3 shows the results of measuring the minimum inhibitory concentration.

Cinnamon essential oil is a liquid Martino showed minimal inhibitory activity in the Bacillus flu 100ppm concentration for pneumoniae (A. Pleuropneumoniae) in the water Toshima, paseuturel La (P. multocida) A and D showed the minimum inhibitory activity at concentrations of 50ppm and 60ppm, respectively. It was confirmed that the result obtained by the liquid medium dilution method shows better activity than the result of measuring the inhibition concentration by the above agar diffusion method. This is because diffusion of the antimicrobial substance is better in the liquid medium than solid agar.

Example  4. Determination of the minimum inhibitory concentration of essential components of cinnamon essential oil

Certain ingredients of cinnamon essential oil are known to be effective against A. pleuropneumoniae and Passthrough la Water Toshida ( P. multocida ) To determine whether the growth of A and D was inhibited, the minimum inhibitory concentration was measured for the major components of cinnamon essential oil. Cinnamon essential oil consists of more than 90% cinnamaldehyde and the rest is ethyl cinnamate, eugenol (usually on leaves), beta-caryophyllene, Linalool, and methyl chavicol. Cinnamaldehyde was the major component of cinnamon essential oil, which was purchased from Sigma Aldrich (St. Louis, Mo., USA). The measurement was performed by modifying the liquid medium dilution method in the same manner as the measurement of the minimum inhibitory concentration of the cinnamon essential oil. DMSO was used as a negative control and antibiotic ampicillin was used as a positive control. The measurement results are shown in Table 4.

The minimum inhibitory concentration of cinnamaldehyde on Actinobacillus fluro- pneumoniae was 1000 ppm, and the rest of the pastelolactone The strains A and D showed no inhibitory activity at the same concentration. The minimum inhibitory concentration of cinnamaldehyde decreased about 10 times the minimum inhibitory concentration of cinnamon essential oil.

The above results suggest that the antimicrobial activity of cinnamon essential oil has a strong antimicrobial activity when it is combined with other minor components in combination.

Claims (9)

An antimicrobial composition for inhibiting bacterial pathogenic bacteria containing cinnamon essential oil as an active ingredient. The antimicrobial composition according to claim 1, wherein the livestock pathogenic bacterium is a bacterium that induces Porcine respiratory disease complex. 3. The method according to claim 2, wherein the bacteria causing the porcine respiratory tract complex infection is Actinobacillus < RTI ID = 0.0 > pleuropneumoniae , < / RTI > Pasteurella multocida ) A and pastelorrhizosta D. The antimicrobial composition for inhibiting bacterial pathogenic bacteria according to claim 1, A method for preventing or treating pathogenic bacterial diseases caused by livestock comprising the step of treating livestock with an antimicrobial composition for inhibiting livestock pathogenic bacteria according to any one of claims 1 to 3. 5. The method of claim 4, wherein the animal is a pig. 5. The method according to claim 4, wherein the pathogenic bacterium-derived disease of the livestock is a porcine respiratory tract complex infection. A livestock feed additive comprising an antimicrobial composition for inhibiting bacterial pathogenic bacteria according to any one of claims 1 to 3 as an active ingredient. The animal feed additive of claim 7, wherein the animal is a pig. A livestock feed comprising the livestock feed additive of claim 7.

Images