KR20010058414A - The composition of antibacterial complex for animal - Google Patents

Abstract

PURPOSE: A complex antibiotic composition for animal containing iron, aminoglycosides antibiotics and chitosan-oligosaccharide is provided, which is useful for treatment and prevention of domestic animal's bacterially caused disease and anemia, can be used instead of expensive antibiotics, so that it saves production cost and minimizes residue of medicine. CONSTITUTION: A process for the preparation of complex antibiotic composition for animal comprises the steps of: mixing iron(iron dextran) and aminoglycosides antibiotics such as Gentamicin(or Streptomycin, Kanamycin, Amikacin, Spectinomycin, Neomycin) in the weight ratio of 20-40:1 to get Gentamicin mixed ferrous(GMF); mixing low molecular chitosan-oligosaccharide(OCHT) and the GMF in an adequate ratio to improve administration effect of the iron and aminoglycosides antibiotics, and gaining the complex antibiotic composition for animal.

Description

The composition of antibacterial complex for animal

The present invention relates to a composite antimicrobial composition for animals, and more particularly, an aminoglycoside-based mixed agent and a chitosan oligosaccharide comprising iron and an aminoglycoside antibiotic in a weight ratio of 20 to 40: 1. It is contained in a weight ratio of 1, which is effective for the treatment and prevention of bacterial diseases and anemia in animals. It relates to a composite antimicrobial composition for animals showing anemia and disease prevention, and growth promoting effect of the animal.

Penicillin, cephalosporin, macrolide, tetracycline, aminoglycoside, chloramphenicol, polypeptide, polyene, diterpen, Many antimicrobials, including other antibiotics, are currently in use. However, r-plasmid, known as a resistance to most synthetic antimicrobial agents, is widely distributed in Gram-negative gut bacteria such as Escherichia coli and expresses resistance as a genetic factor of chromosomes. Therefore, with the emergence of resistant strains to the antimicrobial agents, a new antimicrobial substance having a stronger antimicrobial activity is developed or used, or an existing antimicrobial agent is mixed and used. However, there are many problems such as excessive cost of medication, overdose of medication, drug residue in livestock products, and the emergence of new resistant bacteria.

At present, the global research direction is focused on the development of non-toxic, non-toxic therapeutics using natural materials [Larry et al., 1989; Oram and reiter, 1968; Ashton et al., 1968]. This broadens the antimicrobial range by mixing with non-toxic, non-toxic preparations using natural natural substances with antimicrobial properties, and extends the range of drug use in the direction to solve residual problems while controlling indiscriminate use by mixed use of drugs. I need to go.

On the other hand, in the case of livestock that are collectively reared, even when the body itself is exposed to the external environment during the outbreak of the disease, it is very difficult to receive appropriate protection and management individually. Therefore, the use of natural natural substances that have antimicrobial and immunity-enhancing effects on the disease of the animal itself enhances immunity to the disease, thereby enhancing the prevention effect against disease infection and promoting growth. You can also expect the effect.

Therefore, the use of natural natural substances with antimicrobial action against the disease of the livestock and its immune system as an antimicrobial agent in the livestock can shorten the recovery period from the disease and shorten the treatment period. Increasing the cost of administration and the problem of drug retention due to long-term use may be solved to some extent, and ultimately, it may be a method to replace the use of conventional antibiotics.

In using the complex antimicrobial agent as described above, the most important thing is to select a natural substance having an immune enhancing effect and antimicrobial activity, and then to determine the appropriate compounding ratio between the antimicrobial agent or other active ingredients and natural interactions. From this point of view, the chitosan oligosaccharide to be used in the present invention has antimicrobial and immune enhancing effects, and when used in combination with other active ingredients, its synergistic effect will be very remarkable.

The chitosan-oligosaccharide (hereinafter referred to as "OCHT") refers to low molecular weight chitosan made by hydrolyzing chitosan produced by deacetylating chitin. These chitin and chitosan are non-toxic [Allan et al, 1979] and are naturally occurring polymer cations that are involved in the synthesis and degradation of organisms but do not cause environmental pollution. Therefore, in recent years, research is being conducted to use these materials as potential resources, for example, environmental wastewater coagulants [Castellanos Perez, 1988], heavy metal adsorbents [Evans and Kent, 1962], It is widely applied in various industries such as seed coating [Hadwiger et al, 1961] and wound treatment [Hadwiger et al, 1965]. In addition, chitosan oligosaccharides have excellent antibacterial effect [長 洋 隆, 1980] and immune enhancing effect [Tanigawa et al, 1992], and are widely used for the prevention and treatment of diseases. However, since chitosan is a very high molecular material and is limited in absorption and use in vivo, low molecular weight chitosan oligosaccharides made by hydrolyzing such chitosan are currently used as functional agents.

In addition, the antimicrobial mechanism of chitosan claims that the amino group of chitosan specifically inhibits bacterial growth by binding to the cell wall of the pathogen [Uchida, 1995] and the effect of chitosan's antimicrobial activity on general bacteria on the cell surface structure. [Young et al, 1992] and the claim that it inhibits DNA formation during the metabolic process of bacteria [Stossel and Leuba, 1984], although it is not yet clearly identified, it is known to be particularly effective against intestinal microbial pathogens.

On the other hand, regarding the immune activity of chitosan, Matheson et al. (1984) reported that D-glucosamine, the basic unit of chitosan, increases the activity of natural killer cells involved in immune defense. (1986) reported that hexaglucosamine (Glucosamine) in the hydrolyzate of chitosan exhibits high immune activity by reviving or increasing the immune system correlated with macrophages, lymphocytes and cytokines. In addition, the first reported use of chitosan as feed for normal animals was by Landes and Bough. They added 2.15% chitosan for the purpose of precipitating cheese whey solids, then obtained a chitosan-whey precipitate that was fed to rats and found any differences compared to rats fed casein or simple whey solids. The rats were fed with 1.0, 2.5, 5.0% chitosan instead of cellulose, and no specific results were found.

On the other hand, recent bacterial infectious diseases in animals have formed resistance to various drugs, and domestic livestock farmers tend to administer antibiotics indiscriminately, and development and application of new antibiotics in industrial animals is very difficult in Korea and abroad. It is true.

In addition, although each of the compositions used in the present invention is a substance used in a number of clinical fields, these are all used alone, and pharmacological studies on mixed formulations in which they are mixed together as an active ingredient have been made at home and abroad. It is rarely done.

In order to solve the above problems, the inventors of the present invention have many years in order to determine an optimal compounding ratio that shows a synergistic therapeutic effect for the selection of natural substances having antimicrobial activity and immune enhancing effect and mixed use with other active ingredients. As a result of research, when the aminoglycoside-based mixture of iron and aminoglycoside antibiotics is mixed with chitosan oligosaccharides at an appropriate ratio of chitosan oligosaccharides (OCHT), they show synergistic antimicrobial activity and growth promotion effect of piglets. By confirming, this invention was completed.

Accordingly, the present invention provides a composite antimicrobial composition for animals comprising a pharmaceutically effective amount of an aminoglycoside-based mixed agent and chitosan oligosaccharides in which the weight ratio of iron and aminoglycoside antibiotics is 20 to 40: 1. have.

It is another object of the present invention to provide a use of the complex antimicrobial composition for animals as an agent for preventing or treating bacterial infections and anemia in animals, or as a growth accelerator.

The present invention relates to an animal complex antimicrobial composition comprising an aminoglycoside-based mixed agent in which iron and aminoglycoside antibiotics are mixed in a weight ratio of 20 to 40: 1 and chitosan oligosaccharides in a 1: 5 to 5: 1 weight ratio, It is characterized by the use of such a mixed antimicrobial composition as an agent for preventing or treating bacterial infections and anemia in animals, or as a growth accelerator.

On the other hand, the animal in the present invention includes livestock such as chickens, pigs, cattle, horses, dogs, ducks, goats and sheep.

The present invention will be described in more detail as follows.

The present invention comprises an aminoglycoside-based mixed agent in which iron and aminoglycoside antibiotics are mixed in a weight ratio of 20 to 40: 1 and chitosan oligosaccharide (OCHT) in a compound antimicrobial composition for animals in an appropriate blending ratio, economically and pathogenic. The present invention relates to a novel animal complex antibiotic composition having excellent antimicrobial activity against strains, particularly resistant E. coli. In particular, the use of chitosan oligosaccharides used in the present invention can not only reduce the rapid recovery and treatment period of the axis by the antibacterial action and immune enhancing action during disease occurrence. The mixing and overuse of various antimicrobial agents, the increase in the cost of dosing due to long-term use, and the residual problem of drugs can be solved to some extent, and ultimately, the use of antimicrobial agents can be replaced. Such chitosan oligosaccharides are characterized by improving the administration effect of iron and aminoglycoside antibiotics used in the composite antimicrobial composition for animals.

Next, iron, aminoglycoside antibiotics and chitosan oligosaccharides contained in the mixed antimicrobial composition of the present invention will be described in more detail.

First, iron used in the present invention is not only responsible for supplying oxygen as a central molecule of red blood cells, but also an element related to oxidation, which is a mechanism of generating energy. Therefore, the growing livestock of the livestock is active blood cell production, iron supply shortage will cause anemia. This anemia can occur even if there is a lack of ingredients such as riboflavin, vitamin B ₁₂ , niacin, folic acid, vitamin C, tryptophan, and methionine. For example, the problem of anemia is important in young pigs, when the mothers do not have enough iron, early delivery, and too many cubs. That is, in the case of animals breeding two or more, it is difficult to supply iron depending on the individual who has anemia, and iron is preventively supplied at an early age. In combination with iron and chitosan oligosaccharides, which are anti-microbial and immune-boosting effects against childhood iron and diseases, which may be insufficient in childhood, it may be a way to prevent young animals from anemia and disease. In addition to showing growth-promoting effects, the use of antibiotics can be reduced, which means that farms can reduce production costs. Such toxicity to iron is known to be relatively safe, but all animals are potentially susceptible. Toxic symptoms include acute symptoms and subacute symptoms. First, acute acute symptoms are characterized by sudden death within minutes to several hours after injection, similar to hypersensitivity reactions. Second, subacute symptoms are accompanied by severe depression and lethargy, leading to death, which is related to the toxic effects of iron overdose. Most livestock do not have iron excretion mechanisms, so iron poisoning depends on the amount of iron already present in the body. This is why some animals die or show no symptoms when the same amount is administered. Iron poisoning occurs most frequently by intravenous administration, followed by intramuscular administration and oral administration. Therefore, the proper use of iron, not excessively, can prevent such side effects.

In particular, the iron used in the present invention can be used to select from iron metals and iron-containing materials, iron-containing materials to help the production of blood cells of young livestock growing in synergy with other antibiotics Meet the iron supply shortage. Therefore, in the present invention, iron dextran selected from iron-containing materials, which are currently used most frequently in livestock, is mixed with aminoglycoside antibiotics in a specific weight ratio and then mixed with chitosan oligosaccharides.

In addition, the aminoglycoside antibiotics used in the present invention and the kinds belonging to these ranges all have similar pharmacological action, antibacterial range and toxicity. The aminoglycoside antibiotic was isolated from Streptomyces griseus fungi, which was discovered by Waksman in 1944, and streptomycin was isolated to become the aminoglycoside agonist. In 1949, neomycin was also isolated from Streptomyces fradiae by Waxman . These aminoglycoside antibiotics are a series of antibiotics belonging to the basic oligosaccaride group composed of amino acids and sugars, all of which exhibit bactericidal activity by inhibiting bacterial protein synthesis. Such aminoglycoside antibiotics are commonly called basic water soluble antibiotics because of their high water solubility as basic substances. Aminoglycoside antibiotics show most antimicrobial effects against gram-negative pathogenic bacteria, showing a clear effect on digestive diseases caused by intestinal pathogenic microorganisms in animals, whereas gram-positive bacteria have weaker effects on streptococci and pneumococci. Bacteria have no antimicrobial activity. In addition, the resistance to susceptible bacteria is another weak point of the aminoglycoside antibiotics, such resistance is due to the r-plasmid of enterobacteria and plasmids of staphylococci, so that aminoglyciside modifying enzyme (aminoglyciside modifying) It is due to the action of enzyme). In addition, most of the resistance due to chromosome variation is due to the change of ribosomes. In addition, there is resistance by lowering or inhibiting the intracellular permeability of aminoglycosides. In addition, the major side effects caused by aminoglycoside antibiotics include nephropathy and eighth cranial nervous system disorders. Therefore, care should be taken to prevent the occurrence of toxicity and side effects when the dosage is excessive or used for a long time. As described above, the aminoglycoside antibiotics commonly used in the veterinary field have various disadvantages of their own, and thus, the development of new substances or supplementation with other antimicrobial substances to compensate for these defects. There is a need for the development of new combination antibiotic compositions that are expected to be synergistic. Representative examples of aminoglycoside antibiotics currently used in the veterinary field include streptomycin, kanamycin, amikacin, gentamicin, spectinomycin, and neomycin. ), And the like.

The aminoglycoside antibiotics used in the present invention include gentamicin, which was isolated from micromonospora in 1963, which was obtained from the culture of micromonospora purpurea . It is an aminoglycoside antibiotic. Strictly speaking, aminoglycosides are separated from streptomysis by using Y for mycin, and those isolated from micromonospora by mi instead of y. In addition, gentamicin usually acts as a bactericidal mechanism, although irreversible binding of the 30S ribosomal ribosomal subunit in susceptible bacteria leads to misreading of the mRNA and eventually with bacteria. Unnecessary proteins are synthesized by inhibiting the protein synthesis of the bacteria themselves, thereby preventing the bacteria from propagating. In addition, gentamicin may be applied parenterally or topically in a mixed form with various antibiotics, but cannot be used orally due to its weak absorption in the gastrointestinal tract. This gentamicin has a similar antimicrobial range to other aminoglycoside antibiotics and shows a particularly good effect upon eye infection by Pseudomonas aeruginosa . And, gentamycin acts on the kinds of nearly all Staphylococcus genus (Staphylococcus), streptococci (Streptococcus), Pseudomonas species (Pseudomonas), in Proteus (Proteus), an alkali agent needs in (Alcaligenes).

The aminoglycoside antibiotics are generally effective against many aerobic Gram-negative bacteria and some aerobic Gram-positive bacteria. The resistance of these aminoglycoside antibiotics is probably due to decreased permeability to bacterial cell walls, changes in ribosomal binding sites, Or the presence of a plasmid-mediated resistance factor obtained by conjugation. As described above, the antimicrobial range of the aminoglycoside antibiotics shows most of the antimicrobial effect on aerobic gram-negative pathogenic bacteria, showing excellent efficacy in digestive diseases caused by the intestinal pathogenic microorganisms of animals. Gentamicin also has some cross-resistance with other aminoglycoside drugs. The antibacterial action of gentamicin is effective by showing bactericidal activity against various bacteria susceptible to gentamicin. However, when an animal is infected with a disease, treatment with antibiotics alone is often difficult. One of the causes is that the animals are collectively reared, and when the disease occurs, the animals themselves are exposed to the external environment. It is very difficult to get proper protection and management individually. Toxicity of these aminoglycoside antibiotics, gentamicin, has the following important side effects: renal toxicity, toxicity and neuromuscular blockade. Renal toxicity caused by aminoglycoside antibiotics is caused by nephropathy or acute necrosis of the tubules. It causes hearing impairment and occurs when a lot of medicine enters the inner or lower lymph of the inner ear. Neuromuscular blockade causes paralysis of skeletal muscles, similar to other neuromuscular blockers, resulting in dyspnea and apnea, especially in mesoasthenia, with severe hypocalcemia or shortly after the use of neuromuscular blockers. In particular, aminoglycoside antibiotics pass through the placenta, which can cause kidney damage and deafness in the fetus. The use of aminoglycoside antibiotics can cause overgrowth of non-sensitive bacteria, including fungi. If superinfection occurs during the treatment of aminoglycoside antibiotics, the drug should be discontinued and replaced with other appropriate treatments.

In addition, as another antimicrobial substance used in the present invention, chitosan oligosaccharide (OCHT) has a molecular weight distribution of 200 to 150,000, preferably a distribution of 1 to 16 sugars of 45% or more, and more preferably 200 to molecular weight distribution. It is 99,000 and shows the characteristic that distribution of 1-16 sugar is 55% or more. The chitosan oligosaccharides of the present invention described above exhibit strong antimicrobial activity [see Table 1], such as E. coli, Salmonella, Staphylococcus, which are easily resistant to general antimicrobial agents, and at the same time exhibit an immune enhancing effect. In the immune-boosting effect, as a result of administering 1 mg of chitosan oligosaccharide used in the present invention per mouse, the gamma interferon (IFN-γ) in blood began to increase markedly after 1 hour of administration, and gamma interferon 2 hours after administration. (IFN- [gamma]) increased up to about 6 times.

In general, the degree of deacetylation increases when chitosan is produced from chitin, but the molecular chains of chitin and chitosan are broken, resulting in very low molecular weight, indicating a limit of meltability. In particular, when chitosan is treated for a long time in a high temperature and alkaline solution, while the degree of deacetylation is increased, not only the color of chitosan changes but also the molecular weight drops rapidly. Therefore, as a disadvantage of the polymer chitosan has been reported a lot of physical properties control method. In addition, as a method for preparing chitosan oligosaccharides, a chemical method and an enzymatic method are used. However, the chemical method has a disadvantage in that hydrochloric acid, a reagent, may remain, and thus, a treatment technique is required to completely remove it. There is an economic problem to use expensive chitosan degrading enzyme. Thus, in the present invention, in order to prevent the cleavage of the molecular chain during the production of chitin, the crab shell is finely ground to 2 mm or less, and then the temperature is adjusted to a temperature of 80 to 100 ° C. while adding nitrogen gas thereto, followed by dilution of 0.5 The pretreatment process removes primary calcium by using 0.5N HCl solution and the secondary treatment with 1 ~ 2N HCl solution using 1% NaOH solution at the temperature of 1 ~ 30 ℃ and removes residual calcium and minerals. At the same time, the decrease in viscosity is suppressed, followed by drying with natural light to obtain chitosan having excellent whiteness (molecular weight distribution: 30,012 to 350,448), followed by an enzymatic reaction using a cellulose hydrolase mixture of 90% or more and a viscosity of 2,200 cps. Hereinafter, the chitosan oligosaccharide whose molecular weight distribution is 200-150,000 and whose distribution of 1-16 sugars is 45% or more of the whole is manufactured and used. It is known that there is little toxicity in animals to chitosan oligosaccharides. When ingested 10 g of chitosan per kg body weight of the mouse, no toxicity was observed, and about 18 g of free chitosan and 16 g of chitosan formate. Ingestion daily did not find any particular harmful phenomenon.

On the other hand, in the present invention, in order to identify the interaction between the mixed formulations and chitosan oligosaccharides mixed with iron and aminoglycoside antibiotics described above, and to clarify the correlation between the various mixing ratios, chicken, pigs affected by various bacterial diseases And to determine the antimicrobial effect of each of the antimicrobial agents and mixed preparations containing chitosan oligosaccharides (OCHT) against Gram-positive and negative bacteria isolated from cattle was measured by testing at the minimum inhibitory concentration (Minimum Inhibition Concentration, MIC). In addition, in order to measure the growth-promoting effect of the combined antibiotic composition of iron and aminoglycoside antibiotics and the chitosan oligosaccharide, the iron was injected intramuscularly at 3 and 7 days of piglets, respectively, Through the measuring method, the growth effect of these mixed compositions was measured. As a result, it was confirmed that the mixing ratio of the aminoglycoside-based mixed agent and the natural chitosan oligosaccharide mixed with the iron and aminoglycoside antibiotics described above is contained in the following specific ratios.

That is, in the present invention, a mixture of iron and aminoglycoside antibiotics, preferably gentamicin, is mixed at a weight ratio of 20 to 40: 1 to form an aminoglycoside antibiotic mixture. At this time, when the mixing ratio of iron and aminoglycoside antibiotics is out of the above range, problems such as a decrease in the therapeutic effect will occur.

In particular, the present invention is characterized in that the combination of the aminoglycoside-based mixture of the iron powder and chitosan oligosaccharides (OCHT) prepared from natural natural materials in a specific mixing ratio in the complex antibiotic composition, these mixing ratios It is preferable to mix in 1: 10-10: 1 weight ratio, More preferably, it is 1: 5-5: 1. At this time, in the composite antibiotic composition of the present invention, if the mixing ratio of the aminoglycoside-based mixed agent and chitosan oligosaccharide (OCHT) is out of the above range, the synergistic effect between the two substances is reduced, the antimicrobial activity of the complex antibiotic is lowered and the therapeutic effect May decrease, which is undesirable.

Accordingly, the present invention is to prepare a complex antibiotic composition by mixing the aminoglycoside-based mixed agent and chitosan oligosaccharides (OCHT) mixed with iron as a specific ratio as described above, to expand the antimicrobial range to a single aminoglycoside-based mixed agent As it can suppress the emergence of antibiotic resistant strains, it can be expected to have an excellent effect on the prevention and treatment of diseases when the disease occurs, and also to give the characteristics that can promote growth by preventing anemia of animals. Will be done.

Meanwhile, as mentioned above, since the aminoglycoside antibiotic and iron have their own toxicity, the aminoglycoside-based mixed agent and chitosan oligosaccharide mixed with iron are mixed at a predetermined ratio. In this case, careful application is required because it may cause toxicity in renal or pregnant animals, and subcutaneous administration in animals is considered to be desirable. Otherwise, it acts as a safe combination antimicrobial agent to animals.

On the other hand, the composite antibiotic composition for animals of the present invention, including a pharmaceutically effective amount of an aminoglycoside-based mixed agent and OCHT mixed in a weight ratio of 1: 5 to 5: 1 having a common knowledge in the art to which the present invention belongs. According to a method that can be easily carried out by itself, it may be formulated into a veterinary injection and a liquid. For example, the injectables and solutions for animals containing the complex antimicrobial composition of the present invention include pharmaceutically acceptable carriers such as distilled water, ethyl oleate, ethanol, propylene glycol, glycerin, and the like; Ascorbic acid, sodium hydrogen sulfite, sodium pyrosulfite, tocopherol, etc .; Phenyl mercury nitrate is prepared using chimerosal, benzalkonium chloride, phenol, cresol, methyl paraoxybenzoate, benzyl alcohol, and the like as a preservative.

In addition, the administration of the complex antibiotic composition according to the present invention is administered by the method of intramuscular injection that is commonly applied to animals as an injection, and the liquid is administered in a mixture of oral and negative water. The dosage is 30 to 300 mg / kg body weight of animal for the purpose of treatment of the disease in the case of injection and liquid using the mixed antibiotic composition mixed in the ratio according to the present invention (genamicin of aminoglycosides, iron and chitosan oligosaccharides). Mixed ingredients) is administered based on the condition of the animal.

As described above, the composite antibiotic composition for animals of the present invention is an aminoglycoside-based mixture of natural natural substances having antimicrobial action and immunity strengthening effect on diseases of the animal itself and iron which is easily lacking in the childhood of the animal. Containing the formulation at the same time, it can be used for livestock not only for the rapid recovery from the disease and shorten the treatment period, but also for the mixing and overuse of various antibiotics, the increase of the dosing cost due to the long-term use, and the retention of drugs. The problem may be solved to some extent, and may be another method of ultimately increasing the administration effect of the aminoglycoside-based mixed agent mixed with iron. In addition, young animals can be prevented from anemia and disease, and the growth of the young animals can be reduced and the overuse of antibiotics can reduce the production cost in the farm. Therefore, the combination of these materials is a very realistic method.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it is to those of ordinary skill in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. Will be self-evident.

Production Example

Gentamicin ferrous (GMF) was prepared under general manufacturing laboratory conditions in a weight ratio of 20: 1 to iron (formed as iron dextran in Dextra products, Canada) and gentamicin (purchased from China).

Examples 1-5 and Comparative Examples 1-8

① Antibacterial activity test

The antibiotic susceptibility test medium Müller Hinton agar medium (17.5% hydrolyzed casein with acid, 3% bee extract, 1.5% starch, 15% agar) as shown in Table 1 below. The concentration of the mixture of GMF and OCHT in a mixed ratio was adjusted by two-step dilution. The chitosan oligosaccharides used in the present example [microsomatic trade, "dark horse"] have a chitosan oligosaccharide having a deacetylation degree of 90% or more, a molecular weight distribution of 200 to 99,000, a distribution of 1 to 16 sugars of 55% or more, and a viscosity of 2,200 or less. Used. In addition, the strains used in this example are Streptococcus faculis (A strain) and Staphylococcus aureus R-209 (strain B) as gram-positive strains, and E. coli (Gram negative strain) Escherichia coli ) K88ac (C strain), Escherichia coli K88ab (D strain), Escherichia coli BE (E strain) and Salmonella typhymurium (F strain) were inoculated into the medium and incubated at 37 ° C for 10 to 20 hours. Investigation of the concentration of antibiotics in the lowest concentration medium without the growth of bacteria, determined the minimum inhibitory concentration (MIC) and the results are shown in Table 2 below.

This was in accordance with the National Committee for Clinical Laboratory Standards (NCCLS) method. Each of the strains is a pathogen such as diarrhea, mastitis, and pneumonia of each animal.

As can be seen in Table 2, the composite antibiotic composition for animals of the present invention is 1: 5 to 5: compared with the case of using a combination of iron glycoside antibiotics (GMF) or chitosan oligosaccharides (OCHT) alone. When administered within a specific range of 1 weight ratio, the MIC value was lowered by 16 times, indicating that there was a synergy between the mixture of these aminoglycoside antibiotics mixed with iron and chitosan oligosaccharides.

② Growth Promotion Effect

GMF and chitosan oligosaccharide complex antibiotic composition of 1: 5 to 5: 1 were administered 1 ml each at 3 and 7 days of piglets, and then the average body weight of 26 days was determined to administer only GMF and gutosan oligosaccharides. This was compared with the mean weight of piglets at 26 days of age. The number of experimental heads of each experimental group according to each experimental dose was 30 heads (10 groups were used as one group, and three groups were tested per one experimental group according to each experimental dose). It is shown in Table 3 below.

As can be seen in Table 3, the composite antibiotic composition for animals of the present invention is a specific ratio within the weight ratio of 1: 5 to 5: 1 than when used alone or a mixture of aminoglycoside antibiotics mixed with iron or chitosan oligosaccharides When administered in the ratio, the growth promoting effect of the animals increased by 25 to 31% more than the GMF single dose, and increased by 12 to 17% more than the OCTH single dose, and mixed with aminoglycoside antibiotics mixed with iron Or it can be seen that there is a synergy between oligosaccharides chitosan material.

Therefore, in the present invention, it is preferable that the mixing ratio of the aminoglycoside antibiotics mixed with iron powder or the chitosan oligosaccharide is mixed in a weight ratio of 1: 5 to 5: 1, as shown above. Deviation reduces the synergistic effect between the two substances.

Experimental Example

In order to perform an outdoor treatment test for livestock disease of the complex antibiotic composition of the above embodiment, it was tested by the following composition and method.

Experimental drug 1 (hereinafter referred to as "COGF")

Gentamicin: 5 mg

Iron: 100 mg

-Excipient (5% chitosan oligosaccharide): suitable

Thirty diarrhea piglets within 3 to 10 days of age were selected for E. coli, and 20 ml of the drug administration group and 10 control groups were injected with 1 ml / 10kg of COGF for 3 days. Anatomical and tissue irritation tests for piglets are performed after 3 days of intramuscular injection in piglets, and then the piglets are dissected on the 1st, 3rd, 5th and 7th day to examine respiratory, digestive, hepatic, kidney and injection sites. The muscles were examined and the results are shown in Table 4 below.

As can be seen in Table 4, when using the pharmaceutical composition according to the present invention a significant change was not recognized by the naked eye, but the injection site muscles showed mild hyperemia within 3 days after injection, but was not observed thereafter.

Experimental Example 2

Toxicity test during intramuscular injection and oral administration of the composite antibiotic composition prepared in the above Examples was carried out as follows.

The drug COGF of Experimental Example 1 was used, and the experimental animals were Spraque Dawley rats without specific pathogens supplied from KAIST experimental animal center, and 20 females and 24 females of 5 to 6 weeks of age were purified for 3 days before administration. After that, the animals were reared in a rat cage and fed freely with feed and drinking water.

The experimental group was divided into 0.1ml (recommended amount), 0.3ml (three times the recommended amount), and 0.5ml (five times the recommended amount) per kg body weight, and five oral administration groups were assigned according to the administration route. The intramuscular injection group selected 6 healthy individuals and assigned them. The dose was determined by measuring the weight before administration of the evaluation drug, and the weight before administration was 120-135 kg.

Each experimental group was administered orally with a syringe for intragastric administration for rats for 5 consecutive days once daily for oral administration. Intramuscular injection was given once a day for 3 days. Doses for each subject were based on body weight at each administration, and administration time was administered at 9-10 am each time, and the control group was fed only drinking water and feed.

Autopsy schedule (oral administration group: 1, 5, 10, 14 days after administration; intramuscular injection group: before injection, 1, 3, 5, 9 and 14 days), the weight of the lungs, kidneys, and spleens was measured by ether anesthesia, specific body weights, blood collections, and autopsies immediately followed by partial autopsy procedures. In addition, each experimental organ, including the brain, was extracted and fixed in 10% neutral formalin solution and embedded in paraffin through a partial tissue treatment process, and then cut into 3 to 4 μm for Hematoxylin & Eosin (H & E) staining. , The presence or absence of tissue lesions under an optical microscope and the results are shown in Tables 5-7.

In Table 6, when 0.1 ml was administered, only mild congestion of the intestine was observed, and no pathological lesions were observed in other tissues.

In Table 7, only congestion within the physiological level range was seen, and no change was observed 5 days after the injection.

From the above results, it can be seen that the composite antimicrobial composition of the present invention has no toxicity and excellent stability when the reference dose is used according to the use criteria.

As described above, the present invention provides a composite antibiotic composition for animals comprising an aminoglycoside-based mixed agent and chitosan oligosaccharide mixed with iron in a weight ratio of 20 to 40: 1, which can be used for bacterial infectious diseases and anemia in livestock. It is useful for treatment and prevention, and it is possible to replace expensive new antibiotics, which ultimately reduces the production cost of livestock farms and effectively minimizes the residual of drug in the shaft.

Claims (8)

An animal antimicrobial composition comprising an aminoglycoside-based mixed agent in which iron and an aminoglycoside antibiotic are mixed in a weight ratio of 20 to 40: 1 and chitosan oligosaccharide in a 1: 5 to 5: 1 weight ratio. According to claim 1, wherein said iron is iron dextran (Iron dextran) complex antimicrobial composition for animals, characterized in that. According to claim 1, wherein the aminoglycoside antibiotic is a composite antimicrobial composition for animals, characterized in that the gentamicin (Gentamicin). According to claim 1, wherein the chitosan oligosaccharide has a molecular weight distribution of 200 to 150,000, the distribution of 1 to 16 sugars complex antimicrobial composition for animals, characterized in that the more than 45%. The composition of claim 4, wherein the chitosan oligosaccharide has a molecular weight distribution of 200 to 99,000 and a distribution of 1 to 16 sugars of 55% or more. The complex antimicrobial composition for animals according to claim 1, wherein the animal is at least one livestock selected from the group consisting of chicken, pig, cow, horse, dog, duck, goat and sheep. Preventive or therapeutic agent for anemia, characterized in that it comprises a composite antimicrobial composition for animals of claim 1. Preventive or therapeutic agent for bacterial infections, characterized in that it comprises a composite antimicrobial composition for animals of claim 1.