KR101106053B1 - Antimicrobial composition for pathogenic microorganism comprising plant or extract thereof - Google Patents

Abstract

The present invention relates to an antibiotic composition against a pathogenic microorganism comprising the plant or its extract as an active ingredient, a method for preparing the same, and an application to the feed of the composition.

Description

Antimicrobial composition for pathogenic microorganism comprising plant or extract according to antimicrobial activity against pathogenic microorganisms

The present invention relates to an antibiotic composition for pathogenic microorganisms comprising an extract of a plant as an active ingredient, and more specifically, Citrus aurantifolia swingle (Lime), Citrus aurantiolilia leaf (Citrus) aurantifolia swingle leaf; Lime leaf, Curcuma domestica Valeton; Turmeric, Sesbania grandiflora L. And it relates to an antibiotic composition for pathogenic microorganisms comprising one or more plants and / or extracts thereof selected from the group consisting of Piper sarmentosum Roxb as an active ingredient.

In general, antibiotics play a very important role in the livestock industry as well as in humans. In the past, with the development of livestock raising, the number of heads per unit area has increased and the prevention and treatment of disease has been an important factor for successful livestock raising. Various antibiotics and growth promoters have been developed and used in the field. The increased use of antibiotics has resulted in contamination of livestock products, and furthermore, bioconcentrations have led to life-threatening conditions for human consumption of livestock products.

In addition, the use of antibiotics plays an important role in the development of the livestock industry, but it has caused various side effects such as the generation of resistant bacteria, and the recent increase in the consumption and demand of eco-friendly agricultural products has increased the negative recognition of the use of antibiotics. In fact, the European Union (EU) is tightening restrictions on the use of antibiotics such as allowing only four antibiotics, avilamycin, flavophopholipidol, salinomycin, and monensin. In addition, for the survival of livestock industry in today's fast-changing free trade system, it is important to exclude livestock and to raise livestock in an environmentally friendly manner.

The development of environmentally friendly materials that can replace the function of antibiotics is an important task for the construction of environment-friendly livestock. Eco-friendly antibiotic substitutes have been approached in various fields, and extracts are developed and commercialized by probiotics, which are useful microorganisms that show direct antagonistic action against pathogenic microorganisms, and substances that exhibit antimicrobial activity from plants and other natural substances. It can be divided into fields (Kim Soo Ki. 2005. Use and function of plant extract for feed addition, monthly feed journal).

Useful plant resources, such as herbs, have been applied to disease prevention and treatment because of their direct antibacterial activity, as well as numerous bio-active chemical compounds and various nutrients such as vitamins, calcium and iron.

Perfume plants among various plant resources are generally scented perennial plants have been used as food, fragrance, medicine. In particular, garlic, horseradish, mustard, oregano have been widely used as general foods, ginseng, incense acacia, wormwood, chrysanthemum, irises, gujeolcho, rose, zinnia, cheonilhong, cinnamon, juniper, citron, tanza Perfume plants have been widely used because of their excellent biological activity.

In addition to its physiological activity, plant resources contain substances that exhibit antimicrobial activity against various pathogenic microorganisms. Research into the use of antimicrobial substances in plant resources as an antimicrobial material has been active in food, medicine, and biotechnology industries for a long time. It has been studied. (Shin. DH, Kim. MS, Han. JS 1997. Korean J. Food SCI. TECHNOL . Vol. 29, pp. 808-816; Yang. MS, Ha. YL, Nam. SH, Choi. SU, Jang. DS 1995. Korean SOC . ApplI . Bio . Chem . Vol. 38, pp. 584-589).

There are about 250,000 to 500,000 species of plant resources with antimicrobial and pharmacological activity, but only 1% of them have been evaluated for their antimicrobial and physiological activity (Cowan. MM 1999. Clin. Microbiol. Rev. 14, 564.584.). Therefore, exploration of various plant resources and screening of antimicrobial activity are very valuable fields and can play an important role in the development of antibiotic substitutes.

The present invention solves the above problems, and the object of the present invention as devised by the above necessity is to provide a plant extract that can replace the use of antibiotics.

Another object of the present invention is to provide a use of the feed of the plant or its plant extracts.

In order to achieve the above object, the present invention provides a citrus aurantifolia swingle (Lime), a citrus aurantifolia swingle leaf (Lime leaf), Curcuma domestiga barenton (Curcuma domestica Valeton; Turmeric), Sesbania grandiflora L, and Piper sarmentosum Roxb selected from the group consisting of or an extract of the plant as an active ingredient Provided is an antibiotic composition for pathogenic microorganisms.

In another aspect, the present invention is Citrus aurantifolia swingle (Lime), Citrus aurantifolia swingle leaf (Lime leaf), Curcuma domestica Valeton (Turmeric) ), Sesbania grandiflora L, and Piper sarmentosum Roxb and one or more plants selected from the group consisting of extracting and purifying with water or an organic solvent. It provides a method for preparing an antibiotic composition.

In another aspect, the present invention is Citrus aurantifolia swingle (Lime), Citrus aurantifolia swingle leaf (Lime leaf), Curcuma domestica Valeton (Turmeric) ), Sesbania grandiflora L, and Piper sarmentosum Roxb. Provided are feed compositions comprising one or more plants or extracts thereof.

Hereinafter, the present invention will be described.

The present invention relates to a composition for veterinary or animal husbandry use comprising the active ingredient of the plant of the present invention or an extract thereof mixed in the form of a suitable carrier or feed supplement.

The mentioned compositions can be usefully used in animals, especially livestock such as swine, bovine, ovine, chicken, or equine, without inducing the drawbacks and side effects of antibiotics.

Dosage will depend on a number of factors such as the species, size, weight and age of the animal. In principle, typical dosages will be in the range of 0.1-10 g of extract or plant per animal / day.

Usable extracts according to the invention can be prepared by a process consisting of: a) extracting the plant with a solvent, and optionally b) purifying from water insolubility.

When producing extracts of plants or parts of plants of the invention, water, organic solvents or mixtures thereof can be prepared according to typical methods, but anhydrous distillates can also be applied. Examples of suitable organic solvents are methanol, ethanol, propanol, butanol, methyl acetate, ethyl acetate, methyl ethyl ether, diethyl ether, methylene chloride, chloroform, carbon tetrachloride, benzene, toluene, hexane, petrolatum or acetone. Extraction of the invention also involves the use of such solvent extraction and / or mechanical extraction. The extraction temperature can range from 5 ° C. up to the breaking temperature of the selected solvent.

Natural extracts can be purified by removing insoluble in water (step (b)).

In step (a) the natural extract is concentrated under reduced pressure at 60 ° C. to completely remove alcohol (or any other solvent used among those quoted above) from the solution, and the extract is then extracted from 5% w / w-55% w. / w, preferably to 25% w / w dry residue, and the temperature was maintained at 1 ° C-30 ° C, preferably 4 ° C, without stirring for 1 hour-32 hours, preferably 16 hours.

The resulting clear solution, consisting of purified extracts, was separated from insolubles by centrifugation.

The total amount of ingredient administered to the animal depends on whether the ingredient is an essential compound, an extract of the plant material or a plant material. If the ingredients consist only of plant material, the amount administered is generally from 0.02 to 10 g per kg of body weight per day.

The composition of the present invention may contain components other than the components of the present invention. Suitably the additive contains 0.1 to 100% by weight, preferably 0.2 to 90% by weight, as well as growth promoters, flavorings, absorbent supports and / or other feed ingredients. Preferably, the total amount of growth promoter, fragrance, absorbent support and / or other feed ingredient is 10 to 75% by weight of the additive. If any of the ingredients is a plant material, they are suitably dried and ground into fines or powders before mixing with any other ingredients in the additive.

Examples of suitable growth promoting additives and fragrances are cresol, anetol, decaractone, undecaractone and / or dodecaractone, ionone, ironone, gingerbread, piperidine, propylidene phthalide, boolean Tilidene phthalide, capsaicin and / or tannic acid. The support is for example 40 to 50% by weight wood fibers, 8 to 10% by weight stearin, 4 to 5% turmeric powder, 4 to 5% rosemary powder, 22 to 28% limestone, 1 to It may contain 3% by weight rubber, for example gum arabic, 5-50% by weight sugar and / or starch and 5-15% by weight water. Other feed ingredients are suitably selected from the group consisting of vitamins, enzymes, inorganic salts, ground grains, protein containing ingredients, carbohydrate-containing ingredients, coarse flour and / or bran. The additive is suitably added to the feed composition according to the present invention in an amount such that the feed contains 0.4 ppm to 80% by weight of the component.

Feed compositions according to the invention generally contain the following ingredients as calculated on the basis of the dry weight of the feed:

a) from 0 to 80% by weight of cereals, b) from 0 to 30% by weight of fat, c) from 0 to 85% by weight of anhydrous protein-containing nutrients other than cereals, and d) from 10 ppm to 40% of anhydrous. % By weight of the component of the invention. The total amount of a) to d) is preferably at least 50% by weight anhydrous.

When preparing the feed composition, the additives may be cereals such as milled or ground wheat, oats, barley, corn and rice; Vegetable protein sources based on rapeseed, soybean and sunflower seeds; Animal protein sources; molasses; And dry ingredients consisting of milk products such as various milk powders and whey powders. After mixing with all the dry ingredients, the liquid ingredients and the ingredients which become liquid after heating can be added. The liquid component may consist of lipids, such as fats, for example vegetable fats, and / or carboxylic acids, for example fatty acids, optionally liquefied by heating. After complete mixing, a powder or particulate concentration is obtained depending on the degree of polishing of the components. In order to prevent segregation during storage, water should preferably be added to the animal feed, which is subsequently processed by conventional pelletization, extension or extrusion processes. Any additional water can be removed by drying. If desired, the resulting particulate animal feed can be ground to smaller particle sizes. The disclosed feed compositions are generally administered in combination with dried green grass and / or grasses.

The drinking water supplement may contain at least 1% by weight anhydrous, suitably 1 to 99% anhydrous, preferably 10 to 50% anhydrous. In addition to one or more ingredients, the supplement may contain from 1 to 99 anhydrous weight percent of a greater number of other ingredients.

Suitable examples of other ingredients are mineral salts, vitamins, health and growth improving additives, perfumes, water soluble or water dispersible carriers such as sugars, powdered milk, milk by-products and cellulose derivatives, dispersants and stabilizers such as water soluble Or water-dispersible polymers and mixtures thereof. When preparing drinking water, the replenishment is generally added to the water in an amount such that the concentration of the component is from 1 ppm to 10% by weight.

Within the scope of the present invention, it is also possible to prepare suspensions of the feed composition. This is particularly convenient when the feed is prepared for immediate consumption.

In the present invention, it was carried out to search for useful plant resources with potential to replace antibiotics for the purpose of preventing and treating diseases caused by pathogenic bacteria among antibiotics used in livestock and to investigate the antimicrobial activity spectrum. In particular, we examined the antimicrobial activity spectrum of major pathogenic microorganisms involved in livestock digestive and respiratory diseases among wild plants known for their excellent pharmacological function. In addition, various solvent extraction fractions were obtained for each plant for identification of antimicrobial active substances, and antimicrobial activity spectra were investigated.

In order to evaluate the antimicrobial activity of more detailed plant extracts, hot water extraction method and fractions using various solvents such as methanol, n-hexane, chloroform, ethtyl acetate and butanol were obtained, and Clostridium perfringens Type E, Enterotoxigenic E. coli, Burkholderia sp, The antimicrobial activity of six major pathogenic bacteria, including Pantoea agglomerans, Haemopillus parsuis, and Haemopillus somnus, caused animal disease was evaluated using paper disc methods.

Piper sarmentosum Roxb (Wild Betal) showed weak antimicrobial activity in methanol extract fraction, and Citrus aurantifolia swingle leaf (Lime leaf) showed weak antimicrobial activity in methanol, hexane and chloroform extract fractions. Specific antimicrobial activity was shown. Curcuma domestica Valeton (Turmeric) showed no antimicrobial activity in all solvent extract fractions, but showed specific antimicrobial activity against Clostridium perfringens in hydrothermal extract fractions. Sesbania grandiflora L. did not show antimicrobial activity in most of the extract fractions, but showed antimicrobial activity against Burkholderia sp. In the butanol fraction. Citrus aurantifolia swingle (Lime) showed very good antimicrobial activity in the hot water extract and methanol extract fractions, and showed very good antimicrobial activity compared to other plant extracts.

Sesbania was tested on the MIC (minimum inhibition concentration) of the substances with antimicrobial activity. of grandiflora L (Agati sesbania, Bai Khae) Burkholderia , a respiratory pathogen, from buthanol extract for sp MIC concentration of 135 μg / ml showed the strongest antimicrobial activity.

In the evaluation of antioxidant activity, most plants showed relatively high antioxidant activity, especially Citrus aurantifolia swingle (Lime), Citrus aurantifolia swingle leaf (Lime leaf), Morinda citrifolia L. leaf (Beach mulberry leaf), Piper sarmentosum Roxb (Wild Betal), Curcuma domestica Valeton (Turmeric), Cassia siamea britt (Siamea Cassia) Methnol extract showed high activity.

The result of the synthesis of a series of the present invention Sesbania of grandiflora L (Agati sesbania, Bai Khae) Burkholderia , a respiratory pathogen, from buthanol extract sp . For It showed the strongest antimicrobial activity, and antioxidative activity evaluation showed relatively high antioxidant activity in most plants, especially Citrus aurantifolia swingle (Lime), Citrus aurantifolia swingle leaf (Lime leaf), Piper sarmentosum Roxb (Wild Betal), Curcuma domestica Valeton (Turmeric) The extract derived from the back shows high activity and is highly likely to be developed as an antibiotic substitute.

1 is a photograph of a tropical plant used in the present invention. A.Citrus aurantifolia swingle in photo; B.Citrus aurantifolia swingle leaf; C. Cassia siamea britt; D. Cocos nucifera L .; E.Morinda citrifolia L. leaf .; F. Sesbania grandiflora L .; G, Piper sarmentosum Roxb; H. Curcuma domestica Valeton.
2 shows several extraction steps of tropical plants.

Hereinafter, the present invention will be described in more detail with reference to non-limiting examples. However, the following examples are intended to illustrate the present invention and are not intended to limit the scope of the present invention by the following examples.

Example  1: candidate plant

In order to search for natural plant resources with excellent antimicrobial activity against various pathogens, this study evaluated antimicrobial activity against tropical plants currently growing in the tropics.

Citrus grows in the tropics aurantifolia swingle (Lime), Citrus aurantifolia swingle leaf (Lime leaf,), Morinda citrifolia L. leaf (Beach mulberry leaf), Sesbania grandiflora L. (Agati sesbania,), Piper sarmentosum Roxb (Wild Betal), Curcuma domestica Valeton (Turmeric), Cassia siamea britt (Siamea Cassia) and Cocos nucifera L. (Coconut) was used as a plant resource, each plant used in the present invention is as shown in Figure 1 or Table 1.

 Each plant was washed 2-3 times with distilled water after collection, and dried for 48 hours after removing water with Kimwi tissue, and the dried sample was pulverized using a grinder and used as an extract sample.

Scientific name of plant Common name Citrus aurantifolia swingle Lime Citrus aurantifolia swingle leaf Lime leaf Morinda citrifolia L. leaf Beach mulberry leaf Sesbania grandiflora L. Agati sesbania Piper sarmentosum Roxb Wild betal Curcuma domestica Valeton Turmeric Cassia siamea britt Siamea cassia Cocos nucifera L. Coconut

Table 1 is a name of tropical plants used in the present invention.

Example  2: extract for evaluation of antimicrobial activity and Fraction  pharmacy

Extract preparation of each plant was obtained step by step extraction fraction based on the method of (2005). Detailed extraction fraction recovery method is as follows. After drying, 3 g of each ground plant was mixed with 27 ml of methanol, and extracted by stirring at 150 rpm for about 6 hours at room temperature. The methanol extract was removed using a filter paper (Whatman filter paper No 2) to recover the filtrate. The filtrate was concentrated under reduced pressure and recovered to the final methanol fraction. The recovered methanol fraction was used for the evaluation of some antimicrobial activity, and the rest were solvent fractions sequentially, such as n-hexane, chloroform, ethylacetate, butanol, depending on polarity (FIG. 2).

The methanol fractions concentrated under reduced pressure, tertiary distilled water, and n-hexane were mixed at a ratio of 1: 1, followed by stirring using a voltex mixer. The solvent mixture was allowed to stand at room temperature to induce tube separation, and the n-hexane layer was recovered using pasteur pippet. The recovered n-hexane layer was concentrated under reduced pressure and used for evaluation of antimicrobial activity. The remaining water layer was mixed with tertiary distilled water and chloroform in a ratio of 1: 1 and stirred. After stirring, the mixture was left to induce delamination and the chloroform layer was recovered using pasteur pippet. The recovered chloroform fraction was concentrated under reduced pressure to evaluate the antimicrobial activity. The remaining water layer was mixed with tertiary distilled water and ethylacetate (1: 1), and the ethylacetate layer and the water layer were separated in the same manner as the chloroform fraction recovery method. The ethylacetate fraction was concentrated under reduced pressure. Was used for the evaluation of antimicrobial activity, and the remaining water layer was mixed with tertiary distilled water and butanol (1: 1), and the final butanol fraction was recovered in the same manner as above. The series of methods is as shown in FIG. Each fraction recovered through vacuum concentration was dissolved again with the corresponding solvent and stored at 4 ° C and used for antimicrobial activity evaluation.

Example  3: Evaluation of antimicrobial activity of plant extracts against livestock pathogenic bacteria

Extracts fractionated with various solvents were evaluated for antimicrobial activity against major livestock pathogenic bacteria, and excellent plant extracts were selected.

Example  4: preparation of pathogens

The pathogen used in the present invention was used as a major pathogenic bacterium causing animal digestive diseases and respiratory diseases, and was used by veterinary quarantine officers, and each strain is shown in Table 2.

Each pathogen stored at -70 ° C was streaked in LB agar using sterile platinum and then incubated at 37 ° C for 24 hours. One of the colonies of each pathogenic bacterium that appeared on the surface of LB agar was taken using platinum teeth, and then transferred to LB broth and incubated at 200 rpm for 24 hours in a 37 ° C shake incubator.

Strain name disease Source ^* Clostridium perfringens
Type E Enterotoxemia (NVRQS) Enterotoxigenic  E. coli E. coli infection in pigs and cattle (NVRQS) Burkholderia sp .  Lung infection, or respiratory disease (NVRQS) Pantoea agglomerans
partial Respiratory diseases (NVRQS) Haemopillus parsuis Glaser from pig (NVRQS) Haemopillus somnus Respiratory diseases (NVRQS)

Table 2 is a list of pathogenic bacteria used in the present invention and represents the ^* National Veterinary Research Quarantine Service.

Example  5: Preparation of Stratified Media for Evaluation of Antimicrobial Activity

In order to evaluate the antimicrobial activity of each plant extract fraction, the preparation of the pathogen was used in the medium culture method. The medium layer used in the antimicrobial activity test was prepared by diluting 0.8% agar (Bacto agar, Difco) solution in 10 ml of 15 ml screw cap tubes, sterilizing for 15 minutes at 121 ℃, 50 ℃ in order to prevent the agar of the tube to harden Stored in water bath.

The cultured pathogen was diluted to a final concentration of 1 × 10 ⁷ to 1 × 10 ⁸ cfu / ml, and then inoculated with 100 μl in a sterile 0.8% agar solution tube stored in a water bath at 50 ° C., and stirred using a voltex mixer. It was then poured into LB agar plate and solidified while cooling at room temperature.

Example  6: Evaluation of antimicrobial activity of plant extract fractions

The antimicrobial activity of the plant extract fractions obtained through a series of procedures was evaluated by the paper disc method according to Yun et al. (2005). The fractions extracted for each solvent were absorbed by 25 µl in sterile 6 mm filter paper disk (Whatman AA Discs) and allowed to stand for 30 minutes to evaporate each solvent at room temperature. After each solvent was volatilized, the pathogenic bacteria were placed on the plated medium, and then incubated in a 37 ° C. incubator for 24 to 48 hours. After incubation, growth inhibition ring formed around the paper disc was observed to evaluate the presence and extent of antimicrobial activity.

Example  7: Minimum inhibitory concentration of plant extract fraction ( MIC ) evaluation

The minimum inhibitory concentration (MIC) of each strain was measured by the broth microdilution method as follows. That is, 50 μl of LB was dispensed into 96-well plates, 50 μl of plant extracts were adjusted by two-fold dilution, and the concentration of bacteria was diluted to 10 ⁴ to 10 ⁵ CFU / ml, and 50 μl was added thereto. . After incubation for 16 hours or more at 37 ℃, the absorbance was measured with a microplate reader at 600nm. The concentration of the sample in wells without turbidity was determined as the MIC (minimum inhibition concentration).

Example  8: Evaluation of Antioxidant Activity of Plant Extract Fractions

Antioxidant activity was determined by DPPH photometric assay. After diluting 1/100 by adding 10 μl of plant extract to 990 μl of ethanal, the absorbance was measured at 518 nm after 30 min reaction at room temperature using 0.3 mM DPPH ethanal solution, and antioxidant activity was measured using the following equation.

Antioxidant activity% = 100-[(absorbance of sample minus absorbance of blank) x 100 / absorbance of control]

The result of the above Example is as follows.

The evaluation of the antimicrobial activity of the extracted fraction-specific solvent for the plant 8 grows in tropical species Piper sarmentosum Roxb (Wild Betal), Citrus aurantifolia swingle leaf (Lime leaf), Curcuma domestica Valeton (Turmeric), Sesbania grandiflora L. (Agati sesbania) and Citrus Antimicrobial activity was detected in aurantifolia swingle (Lime), especially Citrus Very good antimicrobial activity was detected in aurantifolia swingle (Lime). But Morinda citrifolia L. leaf (Beach mulberry leaf), Cassia siamea britt (Siamea Cassia) and Cocos nucifera In L. (Coconut), despite the extraction in various solvents, no antimicrobial activity was observed (Table 3).

plant Pathogenic bacteria Solvent Extraction ¹ A B C D E F Piper sarmentosum Roxb Pantoea agglomerans partial - + - - - - Enterotoxigenic E. coli - + - - - - Burkholderia sp . - + - - - - Haemopillus somnus - - - - - - Haemopillus parsuis - ++ - - - - Clostridium perfringens Type E - - - - - - Citrus aurantifolia swingle leaf Pantoea agglomerans partial - - - - - - Enterotoxigenic E. coli - - - - - - Burkholderia sp . - + + + - - Haemopillus somnus - ++ - - - - Haemopillus parsuis - - - - - - Clostridium perfringens Type E - - - - - - Curcuma domestica Valeton Pantoea agglomerans partial - - - - - - Enterotoxigenic E. coli - - - - - - Burkholderia sp . - - - - - - Haemopillus somnus - - - - - - Haemopillus parsuis - - - - - - Clostridium perfringens Type E +++ - - - + - Sesbania grandiflora L. Pantoea agglomerans partial - - - - - - Enterotoxigenic E. coli - - - - - - Burkholderia sp . - - - - - + Haemopillus somnus - - - - - - Haemopillus parsuis - - - - - - Clostridium perfringens Type E - - - - - - Citrus aurantifolia swingle Pantoea agglomerans partial - +++ - - - - Enterotoxigenic E. coli - +++ - - - - Burkholderia sp . + ++ - - - - Haemopillus somnus +++ +++ - - - ++ Haemopillus parsuis - ++ - - - - Clostridium perfringens Type E - - - - - - Morinda citrifolia L. Pantoea agglomerans partial - - - - - - Enterotoxigenic E. coli - - - - - - Burkholderia sp . - - - - - - Haemopillus somnus - - - - - - Haemopillus parsuis - - - - - - Clostridium perfringens Type E - - - - - - Cassia siamea britt Pantoea agglomerans partial - - - - - - Enterotoxigenic E. coli - - - - - - Burkholderia sp . - - - - - - Haemopillus somnus - - - - - - Haemopillus parsuis - - - - - - Clostridium perfringens Type E - - - - - - Cocos nucifera L. Pantoea agglomerans partial - - - - - - Enterotoxigenic E. coli - - - - - - Burkholderia sp . - - - - - - Haemopillus somnus - - - - - - Haemopillus parsuis - - - - - - Clostridium perfringens Type E - - - - - -

Table 3 shows the antimicrobial activity of the solvent extracts of tropical plants against pathogenic bacteria. ¹ solvent fraction: A. water; B. methanol; C. hexane; D. chloroform; E. ethyl acetate F. butanol and antibacterial activity:-, no inhibition (8 mm); +, very slight inhibition (9mm ~ 11mm); ++, moderate inhibition (11mm ~ 13mm); +++, heavy inhibition (13 ~ 17mm) was shown.

1. Extracts and Solvents Fraction  Antimicrobial activity

Citrus , a fragrant plant with a relatively hard waxy layer of leaves Antibacterial activity was shown in methanol, n-hexane and chloroform fractions in solvent extraction fraction of aurantifolia swingle leaf (Lime leaf). However, each antimicrobial activity fraction Burkholderia , the causative agent of respiratory diseases, sp . And Haemopillus It showed specific antimicrobial activity against somnus and no antimicrobial activity against other pathogenic bacteria.

Piper showing broad lobe characteristics sarmentosum As a result of evaluating the antimicrobial activity of the fractions extracted with various solvents against Roxb (Wild Betal), weak antimicrobial activity was detected in methanol extract and no antimicrobial activity was detected in juice and other solvents. Haemopillus, the causative agent of swine glacier disease The antibacterial activity against parasuis was excellent.

Curcuma belonging to the root vegetable domestica Valeton (Turmeric) showed weak antimicrobial activity in the Ethylacetate fraction and no other antimicrobial activity in the other solvent fractions. But Clostridium causes livestock intestinalemia Specific antimicrobial activity was shown only against perfringens Type E strains and the degree of antimicrobial activity was very good.

Sesbania grandiflora L. (Agati sesbania) extract fraction is Burkholderia It showed antimicrobial activity only against sp . However, only very weak activity was shown.

Citrus belonging to the fruit The aurantifolia swingle (Lime) showed a much broader antimicrobial activity spectrum than the other plants and extract fractions. In particular, the fraction extracted by hot water is Haemopillus It showed high antimicrobial activity against somnus , and methanol extract fraction showed high antimicrobial activity against various pathogens. However, Clostridium causes livestock intestinalemia Perfringens Type E did not show antimicrobial activity. Other Morinda citrifola L. leaf, Cassia siamea britt and Cocos All extract fractions of mucifera L. did not show antimicrobial activity.

2. Minimum inhibitory concentrations of plant extract fractions ( MIC ) evaluation

Sesbania was tested on the MIC (minimum inhibition concentration) of the substances with antimicrobial activity. of grandiflora L (Agati sesbania, BaiKhae) Burkholderia , a respiratory pathogen, from buthanol extract for sp MIC concentration of 135 μg / ml It showed the strongest antimicrobial activity (Table 4).

plant Fraction Pathogens Pantoea
agglomerans Enterotoxigenic
E. coli Burkholderia
sp . Haemopillus
somnus Haemopillus
parsuis Clostridium
perfringens ---------- MIC (μg / mL) ---------- Piper sarmentosum Roxb Methanol 2,010 502 1,005 ND 251 ND Citrus aurantifolia swingle leaf Methanol ND ¹ ND 281 2,250 ND ND n -Hexane ND ND 810 ND ND ND Chlororoform ND ND 198 ND ND ND Curcuma domestica Valeton Ethylacetate ND ND ND ND ND 306 Water ND ND ND ND ND 183 Sesbania grandiflora L. Buthanol ND ND 135 ND ND ND Citrus aurantifolia swingle Methanol 1,980 1,980 990 495 990 ND Buthanol ND ND ND 660 ND ND Water ND ND 660 660 ND ND ¹ ND, Not detected

Table 4 shows the minimum inhibitory concentrations of the fractions and extracts of tropical plants against microorganisms, and ND: not detected in the table.

3. Evaluation of Antioxidant Activity of Plant Extract Fractions

Most plants showed relatively high antioxidant activity, especially Citrus aurantifolia swingle (Lime), Citrus aurantifolia swingle leaf (Lime leaf), Morinda citrifolia L. leaf (Beach mulberry leaf), Curcuma domestica Valeton (Turmeric), Cassia siamea britt (Siamea Cassia) Methnol extract showed high activity in the back (Table 5).

Fraction Plant Citrus aurantifolia swingle Citrus aurantifolia swingle leaf Morinda citrifolia L. leaf Sesbania grandiflora L. Piper sarmentosum Roxb Curcuma domestica Valeton Cassia siamea britt Cocos nucifera L Methanol 96.41% 91.77% 82.12% 62.37% 89.85% 91.35% 92.16% 47.20% n -hexane 28.25% 18.18% 29.06% 22.92% 61.21% 14.16% 18.68% 13.68% chloroform 22.58% 15.94% 34.83% 15.77% 94.37% 95.31% 62.98% 15.60% Ethyl acetate 22.56% 18.38% 17.11% 10.71% 88.58% 93.40% 19.39% 32.83% Butanol 25.07% 13.16% 12.15% 10.75% 30.76% 61.52% 15.01% 22.19% water 31.06% 39.54% 21.31% 14.55% 19.68% 25.35% 23.73% 12.75% pressure 23.80% 61.61% 31.82% 93.37% 83.89% 25.49% 23.83% 15.01%

Table 5 shows the antioxidant activity of the solvent extracts of tropical plants

Claims (8)

Burkholderia sp, or Haemophilus somnus, comprising an extract of any one of methanol, hexane, and chloroform extracts of Citrus aurantifolia swingle leaf (Lime leaf) as an active ingredient. Antibiotic pharmaceutical composition for Haemopillus somnus).
delete delete According to claim 1, wherein the composition is an antibiotic pharmaceutical composition, characterized in that for treating or preventing respiratory diseases. Antibiotic against Burkholderia sp or Haemopillus somnus, comprising extracting and purifying Citrus aurantifolia swingle leaf (Lime leaf) with methanol, hexane or chloroform Method for preparing a pharmaceutical composition. delete Burkholderia sp, or Haemophilus somnus, comprising an extract of any one of methanol, hexane, and chloroform extracts of Citrus aurantifolia swingle leaf (Lime leaf) as an active ingredient. Feed composition having an antibiotic effect on Haemopillus somnus).
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