KR20130062187A - The composition of antibacterial complex for animal - Google Patents

Abstract

PURPOSE: A complex antibacterial composition is provided to ensure excellent antibacterial activity against gram-positive bacteria and gram-negative bacteria, and to treat various infections. CONSTITUTION: A complex antibacterial composition contains 10-35wt% of enrofloxacin, 5-25wt% of trimethoprim, and 40-80wt% of sulfamethoxazole. The complex antibacterial composition contains enrofloxacin, trimethoprim, and sulfamethoxazole mixed in a weight ratio of 2:1:5. A drug for treating infectious diseases in an animal digestive system or an animal respiratory system contains the complex antibacterial composition as an active ingredient. The drug is manufactured in the form of powder, a pellet, a granule, an injection, or an ointment. [Reference numerals] (AA) Observation result after injecting the antibacterial composition of an example(%, mucous diarrhea)

Description

The present invention relates to an antibacterial composition for animals,

The present invention relates to a composite antimicrobial composition for animals, and more particularly, to a composition comprising a combination of enrofloxacin, trimethoprim and sulfamethoxazole, The present invention relates to a composite antimicrobial composition for animals having an excellent antimicrobial activity against both Gram-negative bacteria and Gram-positive bacteria, which are pathogenic bacteria such as bacterial digestive diseases and respiratory diseases in animals.

For the purpose of treating various bacterial infectious diseases of animals, it is preferable to use a penicillin, a cephalosporin, a macrolide, a tetracycline, an aminoglycoside, a chloramphenicol, a polypeptide, a polyene, a diterpene, etc. Many antimicrobial agents such as antibiotics are currently being used. However, the r-plasmid, which is known as a resistance factor for most synthetic antimicrobial agents, is a chromosomal genetic factor and is widely distributed in gram-negative intestinal bacteria such as Escherichia coli to express tolerance. Therefore, a new antimicrobial substance having a stronger antimicrobial activity has been developed or used by mixing the existing antimicrobial agents with the emergence of resistant bacteria to the antimicrobial agents.

However, there are many problems such as excessive cost of medication, excessive administration of medication, residual drug problem in livestock product, emergence of new resistant bacteria, and the like.

Since bacterial infections in animals are an environment in which the environment in the housing is compelled to cause multiple infections, high doses of indiscriminate single antibiotics and application of false antimicrobial agents promote the emergence of antibiotic resistant bacteria. Since the use of antibiotics in feed additives is prohibited in 2011, bacterial diseases in livestock are expected to increase sharply. Therefore, in order for Korea to compete with the advanced livestock nations such as the EU, it is necessary to secure competitiveness in animal husbandry, and at the same time, to develop the animal industry by developing animal medicines, especially compound preparations.

The feasibility of concomitant use of antimicrobial agents has been favored for three reasons. The first is the inhibition of the appearance of resistance strains. Second, the antibiotic dose is reduced by concomitant use and the reduction of toxicity by the antibiotic dose. The third is to increase the therapeutic efficacy by using in combination infection (Antibiotic in Laboratory Medicine, 1991, 3rd edition, p432-433). The synergistic mechanism of antibiotics administration is (1) a combination of trimethoprim and sulfamethoxazole, which are representative of continuous inhibition in the pathogenesis of pathogenic bacteria, which act as bacteriostats when used alone When administered together, it acts as a bactericide. Therefore, combination therapy with sulfamethoxazole-trimethoprim has been treated as a single agent and can be considered as a model of hybridoma antibiotic. (2) mecillinam-ampicillin is a typical example of continuous inhibition of cell wall synthesis. (3) When two antimicrobials are coadministered, one antimicrobial agent may affect other antimicrobial agents and promote the entry into pathogenic bacteria. Combined use of aminoglycoside and beta-lactam antibiotics is a good example. (4) Clavulanic acid-penicillin is a typical combination of antibiotic-inhibiting enzymes. (5) Combination of erythromycin and rifampin to prevent the distribution of resistant strains in combination with antimicrobial agents prevents resistance strains from Rhodociccus equi infection of foal. (Antimicrobial therapy in veterinary medicine, 1993, 2nd edition, p8-6, p68-69).

As an example of co-administration of the antimicrobial agent, in Korean Patent Registration No. 2957420, a composite antibiotic composition for animals comprising a pharmaceutically effective amount of norfloxain and tylosin in a ratio of 1: 4 to 4: 1 by weight is used for livestock and It is not only useful for the treatment and prevention of bacterial diseases in fish, but also can replace expensive new antimicrobial substances, ultimately reducing the production cost of livestock farmers and effectively minimizing residues in the stock of livestock. In Korea Patent Registration No. 1047591, cephalexin and enrofloxacin are combined in a weight ratio of 1: 0.25-0.2: 1 to prevent and treat mastitis in animals compared to a single prescription. A combination antibiotic composition for animals having an excellent effect has been proposed.

However, such a composite antimicrobial agent is difficult to apply to diseases or pathogens different from each other due to its different antimicrobial activity against the bacterium. In particular, it is difficult to find a synergistic effect due to the combination of the antimicrobial agents, The development of multiple antimicrobial agents is a constant necessity.

Enrofloxacin is a fluoroquinolone carboxylic acid antibiotic whose chemical name is 1-cyclopropyl-7- (4-ethyl-1-pyrazalinyl) -6-fluoro-1,4-dihydro- 4-oxo-3-quinolone-carboxylic acid is a bactericidal antimicrobial agent that is used only in animals and specifically inhibits DNA supercoiling by inhibiting DNA gyrase (topoisomerase II), which is essential for DNA replication and transcription. E. coli, Pseudomonas aeruginosa, Klebsiella spp., Enterobacter, Campylobacter, Shigella, Salmonella, , Aeromonas, Haemophilus, Proteus, Yersinia, Serratia, Vibrio species, and the like, which are highly effective against pathogenic bacteria such as Escherichia coli, Aeromonas, Haemophilus, Proteus, And can be said to exhibit excellent therapeutic and prophylactic effects. Enlofloxacin is a unique mechanism of action and does not cause cross-resistance with other antimicrobial agents. Since it does not have a beta-lactam ring in its chemical structure, it is stable in beta-lactamase and is also very effective in beta-lactamase-producing strains .

However, a combination antibiotic including enrofloxacin has been proposed in Korean Patent No. 1047591 mentioned above, but it has been proposed that it is effective for treatment and prevention of mastitis in animals. In general, the treatment of general digestive or respiratory diseases However, antibiotics showing synergistic effects with three combined prescriptions have not yet been put to practical use.

On the other hand, countermeasures against various infectious diseases, which are frequently caused by dense breeding, should be given priority to improve swine management. The most serious infectious diseases in livestock are chronic respiratory infections such as digestive tract infections, diarrhea and various pneumonia and atrophic rhinitis. In addition to these clinical infections, chronic consumptive subclinical infections markedly reduce productivity, resulting in enormous economic losses such as delayed shipment days and reduced feed efficiency. Prevention of these diseases is difficult to eradicate diseases in the form of current livestock management, which is being raised collectively.

In the case of bacterial diarrhea, Escherichia coli was the highest in pig digestive diseases examined at the Seoul National University College of Veterinary Medicine for one year in 2009, but did not perform a precise analysis for pathogenic Escherichia coli. Followed by Salmonella infectious disease, pneumoconiosis, and pork loin. In viral gastrointestinal diseases, swine rotavirus infection was the dominant factor. In addition, diarrhea in the middle of pigs' diarrhea leads to decrease in number of reasons, weight loss for weight loss, increase in stress of cause, and diarrhea causes great economic loss due to stagnation of growth, increase of age at delivery, and deterioration of meat quality. Bacterial respiratory disease should be focused on prevention, and bacterial respiratory diseases take chronic progress rather than acute course, resulting in enormous economic losses such as lowered body weight, lower feed efficiency, delayed shipment days, We must cope more thoroughly with its defense. In the winter, respiratory diseases and diarrheal diseases are also prevalent and preventive measures should be taken.

In the field of swine production, a large amount of antimicrobial substance is used in consideration of the above facts. As a result, the use of an intramuscular drug without sufficient clinical and pharmacological and veterinary examination of the vine will result in promotion of resistance to various antimicrobial substances, And various measures to enhance the effect of the build-up can be ended. In general, many resistant strains are resistant to a single antimicrobial therapy, and recent veterinary pharmacological findings are moving towards attempting to kill multi - drug resistant strains by developing a prescription for a compound antimicrobial substance based on scientific evidence. However, careful attention should be paid to the fact that the development of a partial combination may outweigh the disadvantages and lead to side effects.

Thus, the development of a compound antibiotic for animals should be developed in consideration of all aspects of antimicrobial activity, resistant bacteria, and the reduction of side effects. Therefore, a simple combination prescription requires a method of minimizing adverse effects on the use of antibiotic It is difficult to develop.

As a result of long-term research efforts to develop a desirable composite antimicrobial agent according to the problems and development needs of the prior art, enrofloxacin, trimethoprim and sulfamethoxazole are dissolved in a predetermined ratio , The antimicrobial activity exhibits an excellent synergistic effect. Thus, the present invention has been completed.

Accordingly, it is an object of the present invention to provide a composite antimicrobial composition for animals in which enrofloxacin is combined with trimethoprim and sulfamethoxazole.

In order to solve the above-mentioned problems, the present invention provides a composite antimicrobial composition for animals comprising 10-35% by weight of enlofloxacin, 5-25% by weight of trimethoprim and 40-80% by weight of sulfamethoxazole.

The complex antimicrobial composition according to the present invention has excellent antimicrobial activity, and is excellent in antimicrobial activity against Gram-negative bacteria and Gram-positive bacteria, which are pathogenic bacteria of various bacterial diseases in animals, and exhibit excellent activity effects on various infectious diseases of animals.

Particularly, the complex antibacterial composition of the present invention shows excellent effects on gastrointestinal infectious diseases and respiratory infectious diseases among various infectious diseases of livestock.

In addition, the complex antimicrobial agent of the present invention has a well-established toxicity and efficacy against each antimicrobial agent, and is a combined antimicrobial agent having both safety and efficacy, so that it can be safely used in an animal husbandry farm.

1 is a photograph showing the readout of MIC using a microplate reader at 600 nm in Experimental Example 1 of the present invention.
2 is a photograph showing the Broth microdilution method for reading the minimum antibacterial concentration in Experimental Example 1 of the present invention.
3 is a photograph showing MBC read using the microcolony count method in Experimental Example 1 of the present invention.
4 is a photograph showing a Checkerboard method for measuring Fractional Inhibition Concentration (FIC) in Experimental Example 1 of the present invention.
FIG. 5 is a graph showing the results of dynamic experiments after administration of the composition of Example (E: S: T = 2: 5: 1) according to the present invention to experimental pigs.
FIG. 6 is a graph showing the results of a therapeutic effect test for gastrointestinal infectious diseases after administration of a composition of the present invention (E: S: T = 2: 5: 1) according to the present invention to experimental pigs.
FIG. 7 is a graph showing the results of a therapeutic effect test for a respiratory infectious disease after administration of a composition of Example (E: S: T = 2: 5: 1) according to the present invention to experimental pigs.
FIG. 8 is a graph showing clinical experimental results on the symptoms of gastrointestinal infections after administration of the composition of Example (E: S: T = 2: 5: 1) according to the present invention to experimental pigs.
9 is a graph showing the results of clinical tests on the symptoms of respiratory infections after administration of the composition of Example (E: S: T = 2: 5: 1) according to the present invention to experimental pigs.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composite antimicrobial agent for an animal characterized by comprising an antimicrobial composition comprising 10-35% by weight of enrofloxacin, 5-25% by weight of trimethoprim and 40-80% by weight of sulfamethoxazole. More preferably, enlofloxacin: trimethoprim: sulfamethoxazole is mixed in a weight ratio of 2: 1: 5.

According to the present invention, enrofloxacin is a broad-spectrum antibiotic belonging to the quinolone family, inhibiting the DNA gyrase (type 2 topoisomerase) of a bacterium by inhibiting DNA supercoiling and DNA synthesis, and exhibiting bactericidal activity. The concentration can not be reached and the effect can not be exerted. There is a problem of inducing the expression of the resistant bacteria. If it is used too much, it may cause abnormal symptoms such as vomiting and hypersecretion.

In addition, sulfamethoxazole belongs to the sulfonamide system and acts on the metabolism of folic acid, which plays an important role in the synthesis of bacterial DNA, blocking the conversion of PABA (para-aminobenzoic acid) to DFA (dihydrofolic acid) (Nausea, vomiting, diarrhea), central nervous system toxicity (depression, nausea, vomiting, diarrhea) and excessive use of the drug, Headache), facial edema, and the like. Trimethoprim, used in conjunction with sulfamethoxazole, acts on the metabolism of folic acid and inhibits the dihydrofolate reductase to form tetrahydrofolic acid in dihydrofolic acid, As a component of bacteriostatic action, it is used in combination with other antibiotics due to its low antimicrobial activity and mainly used in combination with sulfonamides. However, if it is used too much, nausea, vomiting, headache, (Decreased folate levels) and anemia.

Therefore, in the present invention, it is important to determine the ratio of the appropriate combination prescription as described above to ensure antimicrobial activity and safety.

Meanwhile, the animal additive including the compound antibiotic of the present invention can be prepared by adding commonly used vitamins, saccharides such as glucose and lactose, starch, horse powder, vermiculite, various powders and enzymes in the used amount range.

Further, when formulating the complex antibiotic of the present invention, antioxidants, moisturizers, solvents, and the like may be further included. The antioxidant may be at least one selected from the group consisting of vitamins A, C, E and derivatives thereof, at least one selected from the group consisting of glycerin, vaseline and wax, and at least one selected from the group consisting of olive oil, soybean oil and coconut oil have.

The present invention includes animal drugs for the treatment of gastrointestinal infectious diseases or respiratory infectious diseases of animals containing the above-described complex antimicrobial composition for animals as an active ingredient. The animal medicine containing the complex antibacterial composition for animals of the present invention as an active ingredient can be formulated into powders, solutions, injections and ointments.

The compound of the present invention may be administered in the form of a powder, granules, an injectable preparation for animals, a solution for oral administration, or the like.

In the present invention, when a mixed antibacterial agent is mixed in the above ratio, 200 g to 50 kg, preferably 500 g to 3 kg per ton of feed can be administered.

The complex antimicrobial composition according to the present invention showed excellent results in stability, safety and toxicity tests, and shows excellent results in residual tests, so that it can be very usefully utilized in a state where stability and safety are assured in an animal husbandry farm.

As described above, the present invention can inhibit the appearance of an antibiotic resistance strain by producing three kinds of active ingredients by mixing at the above ratio, and can reduce the toxicity when administered alone, by decreasing the administration dose according to the combination administration . In addition, the antimicrobial activity is enhanced and the antimicrobial range is broader than that of the existing single or two kinds of compound antibiotics, so that it is very effective in the treatment of actual diseases, and particularly effective in the treatment of diseases caused by compound infection. Particularly, the complex antimicrobial composition of the present invention showed excellent effects in bacterial digestive and respiratory infectious diseases of livestock.

EXAMPLES Hereinafter, the present invention will be described in more detail with reference to the following examples, and the results of the antibacterial activity, safety and the like will be confirmed through experimental examples. However, the present invention is not limited to the examples.

Example

(10 to 35% by weight): S (40 to 70% by weight): T (10 to 25% by weight), three antimicrobial agents (Enrofloxacin (E), Sulfamethoxazole Weight ratio) to prepare a composite antibacterial agent composition. This experiment was divided into 4th laboratory experiment for measuring the antimicrobial effect by the interaction between the ingredients of the ingredients and clinical experiment for the purpose of studying the actual efficacy.

Experimental Example 1: Antibacterial effect experiment

1) Materials and Methods

1) -1 antibiotic preparation

The mixture was composed of EST (enrofloxacin 2: sulfamethoxazole 5: trimethoprim 1 weight ratio), ST (sulfamethoxazole 5: trimethoprim 1 weight ratio), and the single agent was enrofloxacin, sulfa Methoxazole and trimethoprim were prepared, and 1 g of each product was taken and stock solution was prepared and stored at -20 ° C in aliquots before use.

1) -2 Test strain

The strains used in this study were separated from poultry and feces from pigs showing respiratory and enteritis symptoms between February, 2009 and March, 2011. They are the Livestock Hygiene Laboratory of Gyeongbuk Province, Chungnam National University Animal Hospital, National Veterinary Science Quarantine Service, They were used by the veterinary hospitals for testing (Table 1).

Cultivation of the bacteria was performed according to the general method of Lenette et al. The test strains were strains related to respiratory and gastrointestinal diseases, which are always troublesome in pig farms. Enterococcus ( E. coli , n = 9), Salmonella spp., Mycoplasma hyopneumoniae , Sensitivity tests were performed on causative bacteria of pneumonia ( Pasteurella multocida ), pleural pneumonia ( Actinobacillus pleuropneumoniae ), and atrophic rhinitis ( Bordetella bronchseptica ).

Actinobacillus pleuropneumoniae Chocolate agar was incubated at 35 ℃ in CO ₂ incubator for 48 hours or 72 hours. NAD (nicotinamide adenine dinucleotide) was added to TSA agar at 40 μg / ml. In particular, Mycoplasma hyopneumoniae used a strain or an established system that is difficult to measure the culture and antimicrobial activity.

Yeast extract, hematocrit and porcine serum were added to the PPLO medium, and the cells were inoculated into Friis medium for 48 hours or 72 hours. Mycoplasma hyopneumoniae is a well - contaminated strain and was used for the experiment after confirmation by PCR according to previously established methods. The remaining strains were inoculated into optimized MHB medium after preincubation with blood culture medium to confirm the growth of bacteria. Quality control strain (QC) was purchased from the Korean Center for Microbiological Conservation. Gram-negative bacteria were cultured in E. coli KCCM 11234 (ATCC 25922) and Gram-positive bacteria ( S. aureus KCCM 40881 (ATCC 29213)) in tryptic soy broth and tryptic soy agar.

Strain name Strain number Dynamics area Ages Symptom Separation organ Escherichia coli (ETEC)
(n = 9) EC-1 Gunyu 50 days Diarrhea, dehydration Feces EC-2 Gunyu 50 days Diarrhea, dehydration Feces EC-3 Gunyu 50 days Diarrhea, dehydration Feces EC-4 Chungnam 50 days Diarrhea, dehydration Feces EC-5 Chungnam 50 days Diarrhea, dehydration Feces EC-6 Old age 30 days diarrhea Feces EC-7 Old age 30 days diarrhea Feces EC-8 Old age 30 days diarrhea Feces EC-9 Old age 30 days diarrhea Feces Staphylococcus aureus ATCC29213 QC-2 Korea Microorganism Conservation Center (Quality strain) Escherichia coli ATCC 25922 QC-1l Korea Microorganism Conservation Center (Quality strain) Salmonella spp.
(n = 10) SA-1 Gunyu 50 days Diarrhea, enteritis Feces SA-2 Gunyu 50 days Diarrhea, enteritis Feces SA-3 Chungnam 50 days Diarrhea, enteritis Feces SA-4 Old age 30 days Diarrhea, enteritis Feces SA-5 Old age 30 days Diarrhea, enteritis Long sample SA-6 Old age 30 days Diarrhea, enteritis Long sample SA-7 Old age 30 days Diarrhea, enteritis Long sample SA-8 Old age 30 days Diarrhea, enteritis Long sample SA-9 Old age 30 days Diarrhea, enteritis Long sample SA-10 Old age 30 days Diarrhea, enteritis Long sample Pasteurella multocida
(n = 12) PM-1 Old age Slaughter Respiratory symptoms lungs lungs PM-2 Old age Slaughter Respiratory symptoms PM-3 Gunyu 120 days Respiratory symptoms Viru PM-4 Gunyu 120 days Respiratory symptoms Viru PM-5 Chungnam 50 days Respiratory symptoms Viru PM-6 Old age 30 days Respiratory symptoms lungs PM-7 Old age 30 days Respiratory symptoms lungs PM-8 Old age 30 days Respiratory symptoms lungs PM-9 Old age 30 days Respiratory symptoms lungs PM-10 Old age 30 days Respiratory symptoms lungs PM-11 Old age 30 days Respiratory symptoms lungs PM-12 Old age 30 days Respiratory symptoms lungs Actinobacillus pleuropneumonia
(n = 9) AP-1 Gunyu 120 days Fever, respiratory symptoms lungs AP-2 Gunyu 120 days Fever, respiratory symptoms lungs AP-3 Gunyu 30 days Fever, respiratory symptoms lungs AP-4 Gunyu 30 days Fever, respiratory symptoms lungs AP-5 Gunyu 30 days Fever, respiratory symptoms lungs AP-6 Gunyu 30 days Fever, respiratory symptoms lungs AP-7 Gunyu 30 days Fever, respiratory symptoms lungs AP-8 Gunyu 30 days Fever, respiratory symptoms lungs AP-9 Gunyu 30 days Fever, respiratory symptoms lungs Mycoplasma hyopneumoniae
(n = 7) MH-1 Old age 60 days Anorexia, fever lungs MH-2 Gunyu 50 days Fever, joint swelling lungs MH-3 Old age 50 days Respiratory symptoms lungs MH-4 Old age 50 days Respiratory symptoms lungs MH-5 Old age 50 days Respiratory symptoms lungs MH-6 Old age 50 days Respiratory symptoms lungs MH-7 Old age 50 days Respiratory symptoms lungs Bodetella bronchiseptica
(n = 9) BB-1 Chungnam 50 days Joint swelling Viru BB-2 Chungnam 50 days Joint swelling Viru BB-3 Pohang 50 days Fever, anorexia Viru BB-4 Andong 35 days Fever, joint swelling Viru BB-5 Old age 30 days Respiratory symptoms Nasal cavity BB-6 Old age 30 days Respiratory symptoms Nasal cavity BB-7 Old age 30 days Respiratory symptoms Nasal cavity BB-8 Old age 30 days Respiratory symptoms Nasal cavity BB-9 Old age 30 days Respiratory symptoms Nasal cavity

<Pig-Derived Strain and Pre-sale Standard Strain>

2) Determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)

     The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were performed as quantitative tests. The minimum inhibitory concentration (MIC) of the antimicrobial substance was determined by the broth microdilution method. The minimum bactericidal concentration (MBC) was determined as the concentration of 99.9% of the bacteria at the concentration above the minimum inhibitory concentration (MIC). All experiments were carried out in accordance with the NCCLS method. In the minimum inhibitory concentration (MIC) experiment, FIC (fraction inhibition concentration) between E and ST was measured using ST (sulfamethoxazole 5: trimethoprim 1 weight ratio) and E (enrofloxacin) After selecting one of the optimal EST mixture ratio, the MIC and MBC of the selected EST mixture were measured, and the MIC and MBC of the E, S, and T components were measured to compare the antibacterial activity.

For the MIC measurement, the broth microdilution method was used to dilute the final concentration of antibiotics to 0.06-64 ㎍ / ㎖. The bacterial solution was adjusted to 0.5 McFarland (10 ⁵ cfu / ml) and diluted 100 times with 50 μl per well. All plates were positive control wells and negative control wells, incubated at 37 ° C for 18-24 hours in aerobic or anaerobic incubation, and then visualized with a microplate reader at the same time to determine the antimicrobial concentration of MIC 90% inhibition) (see Figs. 1, 2 and 3). The FIC (fraction inhibition concentration) between E and ST was determined using the Checkerboard method (see FIG. 4).

3) Fraction index (FIC index) and fractional sterilization concentration index (FBC index) calculation

(MIC) and minimum bactericidal concentration (MBC) of each of EST, ET, ST, E, S and T measured in the above experiment to confirm the combined effect of EST , fractional inhibitory concentration index (FBC) and fractional bactericidal concentration index (FBC index). The calculation method and the criterion for determination are as shown in the following equation (1).

The definition of Equation 1 is as defined in Korean Journal of Diagnostic Imaging 2005, 25 (5): 312-316 and Journal of Antimicrobial Chemothrapy (2002) 49, 275-282.

4) Experimental results

(1) Combined effect of enlofloxacin with sulfamethoxazole and trimethoprim

 (1) -1 FIC index of rofloxacin and sulfamethoxazole + trimethoprim combination

First of all, for the strains of B. bronchoseptica, E. coli (ATCC 25922), P. hemolytica (ATCC 55518), S. aureus (ATCC 29213), S. cholerasuis (ATCC 7001) and S. typhimurium After the combination of E and ST, the FIC index was calculated using the checkerboard method. The interaction and blending ratio of enlofloxacin (E) with sulfamethoxazole + trimethoprim (ST) were calculated and the results are shown in Table 2 below. As shown in Table 2, co-administration of enrofloxacin (E) with sulfamethoxazole + trimethoprim (ST) showed a synergistic action with FIC index of 0.5 to 1 under the strain.

<FIC index of combined E and ST>

ST85, E25: ST75, E30: ST70, E35: ST65, and E40: ST60 ratio in order to investigate the combined effect of enlofloxacin (E) with sulfamethoxazole and trimethoprim , And the respective MICs of the test strains with ST were measured, and the FIC index was calculated using the measured values.

As a result, the FIC index of each of the 5 groups showed a synergy effect of less than 1, and there was no significant difference among the groups (see Tables 2, 3, 4, 5, 6 and 7).

FIC index of E15 / ST85 compound Strain MIC ([mu] g / mL) FIC index Judgment E15 + ST85 EST-E EST-ST E ST B. broncoseptica 0.5 0.075 0.425 0.25 64 0.307 Increase E. coli
(ATCC 25922) 0.0625 0.00937 0.05313 0.25 One 0.091 Increase P. hemolytica
(ATCC 55518) 0.5 0.075 0.425 0.125 0.5 1.450 irrelevant S. aureus
(ATCC 29213) 0.5 0.075 0.425 0.25 2 0.513 Increase S. cholerasuis
(ATCC 7001) 0.03125 0.00469 0.02656 0.03125 0.5 0.203 Increase S. typhimurium
(Outdoor strain) 32 4.8 27.2 32 128 0.363 Increase

FIC index of E25 / ST75 compound Strain MIC ([mu] g / mL) FIC index Judgment E25 + ST75 EST-E EST-ST E ST B. broncoseptica 0.25 0.0625 0.1875 0.25 64 0.253 Increase E. coli
(ATCC 25922) 0.03125 0.007813 0.023438 0.25 One 0.055 Increase P. hemolytica
(ATCC 55518) 0.25 0.0625 0.1875 0.125 0.5 0.875 Increase S. aureus
(ATCC 29213) 0.25 0.0625 0.1875 0.25 2 0.344 Increase S. cholerasuis
(ATCC 7001) 0.03125 0.007813 0.023438 0.03125 0.5 0.297 Increase S. typhimurium
(Outdoor strain) 32 8 24 32 128 0.438 Increase

FIC index of E30 / ST70 compound Strain MIC ([mu] g / mL) FIC index Judgment E30 + ST70 EST-E EST-ST E ST B. broncoseptica 0.5 0.15 0.35 0.25 64 0.605 Increase E. coli
(ATCC 25922) 0.0625 0.01875 0.04375 0.25 One 0.119 Increase P. hemolytica
(ATCC 55518) 0.5 0.15 0.35 0.125 0.5 1.900 irrelevant S. aureus
(ATCC 29213) 0.5 0.15 0.35 0.25 2 0.775 Increase S. cholerasuis
(ATCC 7001) 0.03125 0.009375 0.021875 0.03125 0.5 0.344 Increase S. typhimurium
(Outdoor strain) 32 9.6 22.4 32 128 0.475 Increase

FIC index of E35 / ST65 compound Strain MIC ([mu] g / mL) FIC index Judgment E35 + ST65 EST-E EST-ST E ST B. broncoseptica 0.125 0.04375 0.08125 0.25 64 0.176 Increase E. coli
(ATCC 25922) 0.03125 0.01094 0.02031 0.25 One 0.064 Increase P. hemolytica
(ATCC 55518) 0.125 0.04375 0.08125 0.125 0.5 0.513 Increase S. aureus
(ATCC 29213) 0.25 0.0875 0.1625 0.25 2 0.431 Increase S. cholerasuis
(ATCC 7001) 0.03125 0.01094 0.02031 0.03125 0.5 0.391 Increase S. typhimurium
(Outdoor strain) 32 11.2 20.8 32 128 0.513 Increase

FIC index of E40 / ST60 compound Strain MIC ([mu] g / mL) FIC index Judgment E40 + ST60 EST-E EST-ST E ST B. broncoseptica 0.125 0.05 0.075 0.25 64 0.201 Increase E. coli
(ATCC 25922) 0.03125 0.0125 0.01875 0.25 One 0.069 Increase P. hemolytica
(ATCC 55518) 0.125 0.05 0.075 0.125 0.5 0.550 Increase S. aureus
(ATCC 29213) 0.25 0.1 0.15 0.25 2 0.475 Increase S. cholerasuis
(ATCC 7001) 0.03125 0.0125 0.01875 0.03125 0.5 0.438 Increase S. typhimurium
(Outdoor strain) 32 12.8 19.2 32 128 0.550 Increase

 (1) -2 yen MIC and FIC index of rofloxacin and sulfamethoxazole, trimethoprimmate

The composition ratio of the most suitable EST combination (enrofloxacin 2: sulfamethoxazole 5: trimethoprim 1) was determined using the above test results (Tables 2, 3, 4, 5, 6 and 7) The minimum inhibitory concentration (MIC) and the fractional inhibitory concentration index (FIC index) were measured (see Tables 8 and 9). As a result, the combined effect of enrofloxacin, sulfamethoxazole and trimethoprim (EST) was confirmed to have a synergistic effect due to the fractional inhibition concentration index (FIC index) of less than 1 depending on the strain. These results indicate that EST can exert more effective bactericidal inhibitory ability at a much lower concentration than using a single agent.

The FIC index (primary) of enlofloxacin, sulfamethoxazole, trimethoprim, Strain name Strain
number MIC (ug / ml) FIC
Indices Judgment EST Monolith EST EST
-E EST
-S EST
-T E S T Escherichia coli EC-1 0.25 0.063 0.156 0.031 8 64 64 0.011 Increase EC-2 0.25 0.063 0.156 0.031 0.25 8 4 0.277 Increase EC-3 0.25 0.063 0.156 0.031 0.25 16 4 0.268 Increase EC-4 0.25 0.063 0.156 0.031 0.25 16 2 0.275 Increase EC-5 0.25 0.063 0.156 0.031 0.25 8 2 0.285 Increase EC-6 0.25 0.063 0.156 0.031 16 64 64 0.007 Increase EC-7 0.25 0.063 0.156 0.031 16 64 64 0.007 Increase EC-8 0.13 0.031 0.078 0.016 16 64 64 0.003 Increase EC-9 0.13 0.031 0.078 0.016 16 64 64 0.003 Increase Salmonella spp. SA-1 0.5 0.125 0.313 0.063 One 64 2 0.161 Increase SA-2 0.5 0.125 0.313 0.063 One 64 2 0.161 Increase SA-3 0.5 0.125 0.313 0.063 One 64 2 0.161 Increase SA-4 0.5 0.125 0.313 0.063 One 64 2 0.161 Increase SA-5 0.5 0.125 0.313 0.063 One 64 2 0.161 Increase SA-6 2 0.500 1.250 0.250 4 64 4 0.207 Increase SA-7 2 0.500 1.250 0.250 4 64 4 0.207 Increase SA-8 One 0.250 0.625 0.125 8 64 4 0.072 Increase SA-9 One 0.250 0.625 0.125 8 64 4 0.072 Increase SA-10 One 0.250 0.625 0.125 8 64 4 0.072 Increase Pasteurella multocida PM-1 0.125 0.031 0.078 0.016 0.125 64 4 0.255 Increase PM-2 0.125 0.031 0.078 0.016 0.125 64 4 0.255 Increase PM-3 0.125 0.031 0.078 0.016 0.125 64 4 0.255 Increase PM-4 0.25 0.063 0.156 0.031 0.125 64 4 0.510 Increase PM-5 0.25 0.063 0.156 0.031 0.125 64 4 0.510 Increase PM-6 0.25 0.063 0.156 0.031 2 64 4 0.042 Increase PM-7 0.25 0.063 0.156 0.031 2 64 8 0.038 Increase PM-8 0.25 0.063 0.156 0.031 2 64 8 0.038 Increase PM-9 0.25 0.063 0.156 0.031 2 64 8 0.038 Increase PM-10 One 0.250 0.625 0.125 4 64 8 0.088 Increase PM-11 One 0.250 0.625 0.125 4 64 8 0.088 Increase PM-12 One 0.250 0.625 0.125 4 64 8 0.088 Increase Actinobacillus pleuropneumonia AP-1 0.125 0.031 0.078 0.016 0.5 64 8 0.066 Increase AP-2 0.125 0.031 0.078 0.016 0.5 64 8 0.066 Increase AP-3 0.125 0.031 0.078 0.016 0.5 64 16 0.065 Increase AP-4 0.125 0.031 0.078 0.016 0.5 64 16 0.065 Increase AP-5 0.5 0.125 0.313 0.063 4 64 16 0.040 Increase AP-6 0.5 0.125 0.313 0.063 4 64 16 0.040 Increase AP-7 0.5 0.125 0.313 0.063 4 64 16 0.040 Increase AP-8 0.5 0.125 0.313 0.063 4 64 16 0.040 Increase AP-9 0.5 0.125 0.313 0.063 4 64 16 0.040 Increase Mycoplasma hyopneumoniae MH-1 2 0.500 1.250 0.250 2 64 64 0.273 Increase MH-2 0.25 0.063 0.156 0.031 0.25 64 64 0.253 Increase MH-3 One 0.250 0.625 0.125 One 64 64 0.262 Increase MH-4 One 0.250 0.625 0.125 One 64 64 0.262 Increase MH-5 One 0.250 0.625 0.125 2 64 64 0.137 Increase MH-6 One 0.250 0.625 0.125 2 64 64 0.137 Increase MH-7 One 0.250 0.625 0.125 2 64 64 0.137 Increase Bodetella bronchiseptica BB-1 One 0.250 0.625 0.125 0.25 16 64 1.041 addition BB-2 0.25 0.063 0.156 0.031 0.25 4 64 0.290 Increase BB-3 0.25 0.063 0.156 0.031 2 16 64 0.042 Increase BB-4 0.25 0.063 0.156 0.031 0.25 8 64 0.270 Increase BB-5 0.25 0.063 0.156 0.031 0.25 8 64 0.270 Increase BB-6 0.125 0.031 0.078 0.016 0.125 16 64 0.255 Increase BB-7 0.125 0.031 0.078 0.016 0.125 16 64 0.255 Increase BB-8 0.125 0.031 0.078 0.016 0.125 16 64 0.255 Increase BB-9 0.125 0.031 0.078 0.016 0.125 16 64 0.255 Increase Staphylococcus aureus ATCC29213 QC-2 0.5 0.125 0.313 0.063 One 8 2 0.195 Increase Escherichia coli ATCC 25922 QC-1l 0.125 0.031 0.078 0.016 0.5 8 2 0.080 Increase

The FIC index (second order) of enlofloxacin, sulfamethoxazole, trimethoprim, Strain name Strain
number MIC (ug / ml) FIC
Indices Judgment EST Monolith EST EST
-E EST
-S EST
-T E S T Escherichia coli EC-1 0.5 0.125 0.313 0.063 4 128 4 0.049 Increase EC-2 0.063 0.016 0.039 0.008 0.063 32 4  0.253 Increase EC-3 0.063 0.016 0.039 0.008 0.063 32 4  0.253 Increase EC-4 0.063 0.016 0.039 0.008 0.063 16 2   0.256 Increase EC-5 0.063 0.016 0.039 0.008 0.063 32 2   0.255 Increase EC-6 0.25 0.063 0.156 0.031 4 128 32   0.018 Increase EC-7 0.25 0.063 0.156 0.031 8 128 32   0.010 Increase EC-8 0.25 0.063 0.156 0.031 8 128 32   0.010 Increase EC-9 0.25 0.063 0.156 0.031 8 128 32   0.010 Increase Salmonella spp. SA-1 One 0.250 0.625 0.125 2 128 2   0.192 Increase SA-2 One 0.250 0.625 0.125 One 128 2   0.317 Increase SA-3 One 0.250 0.625 0.125 One 128 2   0.317 Increase SA-4 0.5 0.125 0.313 0.063 One 128 2   0.159 Increase SA-5 0.5 0.125 0.313 0.063 One 128 2   0.159 Increase SA-6 2 0.500 1.250 0.250 4 128 4   0.197 Increase SA-7 2 0.500 1.250 0.250 4 128 4   0.197 Increase SA-8 One 0.250 0.625 0.125 8 128 4   0.067 Increase SA-9 One 0.250 0.625 0.125 8 128 4   0.067 Increase SA-10 One 0.250 0.625 0.125 8 128 4   0.067 Increase Pasteurella multocida PM-1 0.125 0.031 0.078 0.016 0.5 128 4   0.067 Increase PM-2 0.125 0.031 0.078 0.016 0.5 128 4   0.067 Increase PM-3 0.125 0.031 0.078 0.016 0.5 128 4  0.067 Increase PM-4 0.125 0.031 0.078 0.016 0.5 128 4   0.067 Increase PM-5 0.125 0.031 0.078 0.016 0.5 128 4   0.067 Increase PM-6 0.25 0.063 0.156 0.031 0.5 128 4   0.134 Increase PM-7 0.25 0.063 0.156 0.031 2 128 8   0.036 Increase PM-8 0.25 0.063 0.156 0.031 2 128 8   0.036 Increase PM-9 0.25 0.063 0.156 0.031 2 128 8   0.036 Increase PM-10 One 0.250 0.625 0.125 4 128 8   0.083 Increase PM-11 One 0.250 0.625 0.125 4 128 8   0.083 Increase PM-12 One 0.250 0.625 0.125 4 128 8   0.083 Increase Actinobacillus pleuropneumonia AP-1 0.25 0.063 0.156 0.031 0.5 64 2   0.143 Increase AP-2 0.25 0.063 0.156 0.031 0.5 64 2   0.143 Increase AP-3 0.125 0.031 0.078 0.016 0.5 64 2   0.072 Increase AP-4 0.125 0.031 0.078 0.016 0.5 64 2   0.072 Increase AP-5 0.5 0.125 0.313 0.063 2 64 4   0.083 Increase AP-6 0.5 0.125 0.313 0.063 2 64 4   0.083 Increase AP-7 0.5 0.125 0.313 0.063 2 64 4   0.083 Increase AP-8 0.5 0.125 0.313 0.063 2 64 4   0.083 Increase AP-9 0.5 0.125 0.313 0.063 2 64 4   0.083 Increase Mycoplasma hyopneumoniae MH-1 0.25 0.063 0.156 0.031 0.25 64 64   0.253 Increase MH-2 0.25 0.063 0.156 0.031 0.25 64 64   0.253 Increase MH-3 0.25 0.063 0.156 0.031 0.25 64 64   0.253 Increase MH-4 0.25 0.063 0.156 0.031 0.25 64 64   0.253 Increase MH-5 0.5 0.125 0.313 0.063 One 64 64   0.131 Increase MH-6 0.5 0.125 0.313 0.063 One 64 64   0.131 Increase MH-7 0.5 0.125 0.313 0.063 One 64 64  0.131 Increase Bodetella bronchiseptica BB-1 0.125 0.031 0.078 0.016 0.25 64 8  0.128 Increase BB-2 0.125 0.031 0.078 0.016 0.25 64 8   0.128 Increase BB-3 0.125 0.031 0.078 0.016 2 64 8   0.019 Increase BB-4 0.5 0.125 0.313 0.063 0.25 64 8   0.513 Increase BB-5 0.125 0.031 0.078 0.016 2 64 8   0.019 Increase BB-6 0.125 0.031 0.078 0.016 2 64 16   0.018 Increase BB-7 0.125 0.031 0.078 0.016 2 64 16   0.018 Increase BB-8 0.125 0.031 0.078 0.016 2 64 8   0.019 Increase BB-9 0.125 0.031 0.078 0.016 2 64 8   0.019 Increase Staphylococcus aureus ATCC29213 QC-2 0.5 0.125 0.313 0.063 0.5 32 4   0.275 Increase Escherichia coli ATCC 25922 QC-1l 0.125 0.031 0.078 0.016 0.125 16 2  0.263 Increase

(1) -3 yen MBC and FBC index of rofloxacin, sulfamethoxazole, trimethoprim,

The minimum bactericidal concentration (MBC) and fraction sterilization concentration index (FBC index) of enrofloxacin, sulfamethoxazole and trimethoprim (EST) in the second experiment are shown in the table below Reference). As a result, the combined effect of enrofloxacin, sulfamethoxazole and trimethoprim (EST) was confirmed to be synergistic because the fractional sterilization concentration index (FBC index) was less than 0.5 in almost all strains. These results indicate that EST can exert a more effective sterilization ability at a much lower concentration than using a single agent. The increase in bactericidal potency, as indicated by the FBC index of the compound, is higher than the increase in bacterial inhibition as indicated by the FIC index in actual practice. That is, it is possible to shorten the treatment period and to more reliably inhibit the expression of resistant bacteria.

The FBC index (primary) of enlofloxacin, sulfamethoxazole, trimethoprim, Strain name Strain
number MBC (ug / ml) FBC
Indices Judgment EST Monolith EST EST
-E EST
-S EST
-T E S T Escherichia coli EC-1 0.5 0.125 0.313 0.063 64 64 64  0.008 Increase EC-2 0.50 0.125 0.313 0.063 64 64 64  0.008 Increase EC-3 0.50 0.125 0.313 0.063 64 64 64  0.008 Increase EC-4 0.50 0.125 0.313 0.063 64 64 64  0.008 Increase EC-5 0.50 0.125 0.313 0.063 64 64 64  0.008 Increase EC-6 0.5 0.125 0.313 0.063 64 64 64  0.008 Increase EC-7 One 0.250 0.625 0.125 64 64 64  0.016 Increase EC-8 0.25 0.063 0.156 0.031 64 64 64  0.004 Increase EC-9 0.5 0.125 0.313 0.063 64 64 64  0.008 Increase Salmonella spp. SA-1 2 0.500 1.250 0.250 8 64 16  0.098 Increase SA-2 2 0.500 1.250 0.250 8 64 16  0.098 Increase SA-3 2 0.500 1.250 0.250 8 64 16  0.098 Increase SA-4 2 0.500 1.250 0.250 8 64 16  0.098 Increase SA-5 2 0.500 1.250 0.250 8 64 16  0.098 Increase SA-6 4 1,000 2.500 0.500 16 64 64  0.109 Increase SA-7 4 1,000 2.500 0.500 16 64 64  0.109 Increase SA-8 4 1,000 2.500 0.500 16 64 64  0.109 Increase SA-9 4 1,000 2.500 0.500 16 64 64  0.109 Increase SA-10 4 1,000 2.500 0.500 16 64 64  0.109 Increase Pasteurella multocida PM-1 0.5 0.125 0.313 0.063 4 64 32  0.038 Increase PM-2 0.5 0.125 0.313 0.063 4 64 32  0.038 Increase PM-3 0.5 0.125 0.313 0.063 4 64 32  0.038 Increase PM-4 0.5 0.125 0.313 0.063 4 64 32  0.038 Increase PM-5 0.5 0.125 0.313 0.063 4 64 32  0.038 Increase PM-6 0.5 0.125 0.313 0.063 8 64 32  0.022 Increase PM-7 0.5 0.125 0.313 0.063 8 64 64  0.021 Increase PM-8 2 0.500 1.250 0.250 8 64 64  0.086 Increase PM-9 2 0.500 1.250 0.250 8 64 64  0.086 Increase PM-10 2 0.500 1.250 0.250 8 64 64  0.086 Increase PM-11 2 0.500 1.250 0.250 8 64 64  0.086 Increase PM-12 2 0.500 1.250 0.250 8 64 64  0.086 Increase Actinobacillus pleuropneumonia AP-1 0.5 0.125 0.313 0.063 2 64 64  0.068 Increase AP-2 0.5 0.125 0.313 0.063 2 64 64  0.068 Increase AP-3 0.5 0.125 0.313 0.063 2 64 64  0.068 Increase AP-4 0.5 0.125 0.313 0.063 2 64 64  0.068 Increase AP-5 One 0.250 0.625 0.125 8 64 64  0.043 Increase AP-6 One 0.250 0.625 0.125 8 64 64  0.043 Increase AP-7 One 0.250 0.625 0.125 8 64 64  0.043 Increase AP-8 One 0.250 0.625 0.125 8 64 64  0.043 Increase AP-9 One 0.250 0.625 0.125 8 64 64  0.043 Increase Mycoplasma hyopneumoniae MH-1 2 0.500 1.250 0.250 4 64 64  0.148 Increase MH-2 2 0.500 1.250 0.250 4 64 64  0.148 Increase MH-3 2 0.500 1.250 0.250 4 64 64  0.148 Increase MH-4 2 0.500 1.250 0.250 4 64 64  0.148 Increase MH-5 2 0.500 1.250 0.250 4 64 64  0.148 Increase MH-6 2 0.500 1.250 0.250 4 64 64  0.148 Increase MH-7 2 0.500 1.250 0.250 4 64 64  0.148 Increase Bodetella bronchiseptica BB-1 2 0.500 1.250 0.250 2 64 64  0.273 Increase BB-2 2 0.500 1.250 0.250 2 64 64  0.273 Increase BB-3 One 0.250 0.625 0.125 4 64 64  0.074 Increase BB-4 One 0.250 0.625 0.125 One 64 64  0.262 Increase BB-5 One 0.250 0.625 0.125 One 64 64  0.262 Increase BB-6 One 0.250 0.625 0.125 One 64 64  0.262 Increase BB-7 One 0.250 0.625 0.125 2 64 64  0.137 Increase BB-8 One 0.250 0.625 0.125 One 64 64  0.262 Increase BB-9 One 0.250 0.625 0.125 2 64 64  0.137 Increase Staphylococcus aureus ATCC29213 QC-2 2 0.500 1.250 0.250 2 64 64  0.273 Increase Escherichia coli ATCC 25922 QC-1l One 0.250 0.625 0.125 2 64 64  0.137 Increase

The FBC index (secondary) of enlofloxacin, sulfamethoxazole, trimethoprim, Strain name Strain
number MBC (ug / ml) FBC
Indices Judgment EST Monolith EST EST
-E EST
-S EST
-T E S T Escherichia coli EC-1 0.5 0.125 0.313 0.063 8 128 128   0.019 Increase EC-2 0.5 0.125 0.313 0.063 0.25 128 128   0.503 Increase EC-3 0.5 0.125 0.313 0.063 0.25 128 128   0.503 Increase EC-4 0.5 0.125 0.313 0.063 0.25 128 128   0.503 Increase EC-5 0.5 0.125 0.313 0.063 0.25 128 128   0.503 Increase EC-6 0.5 0.125 0.313 0.063 16 128 128   0.011 Increase EC-7 One 0.250 0.625 0.125 16 128 128   0.021 Increase EC-8 0.25 0.063 0.156 0.031 16 128 128   0.005 Increase EC-9 0.5 0.125 0.313 0.063 32 128 128   0.007 Increase Salmonella spp. SA-1 2 0.500 1.250 0.250 4 128 128   0.137 Increase SA-2 2 0.500 1.250 0.250 4 128 128   0.137 Increase SA-3 2 0.500 1.250 0.250 4 128 128   0.137 Increase SA-4 2 0.500 1.250 0.250 4 128 128   0.137 Increase SA-5 2 0.500 1.250 0.250 4 128 128   0.137 Increase SA-6 4 1,000 2.500 0.500 8 128 128   0.148 Increase SA-7 4 1,000 2.500 0.500 8 128 128   0.148 Increase SA-8 4 1,000 2.500 0.500 16 128 128   0.086 Increase SA-9 4 1,000 2.500 0.500 16 128 128   0.086 Increase SA-10 4 1,000 2.500 0.500 16 128 128   0.086 Increase Pasteurella multocida PM-1 0.5 0.125 0.313 0.063 4 128 128   0.034 Increase PM-2 0.5 0.125 0.313 0.063 4 128 128   0.034 Increase PM-3 0.5 0.125 0.313 0.063 4 128 128   0.034 Increase PM-4 0.5 0.125 0.313 0.063 4 128 128   0.034 Increase PM-5 0.5 0.125 0.313 0.063 4 128 128   0.034 Increase PM-6 One 0.250 0.625 0.125 8 128 128   0.037 Increase PM-7 0.5 0.125 0.313 0.063 8 128 128   0.019 Increase PM-8 2 0.500 1.250 0.250 8 128 128   0.074 Increase PM-9 2 0.500 1.250 0.250 8 128 128   0.074 Increase PM-10 2 0.500 1.250 0.250 8 128 128   0.074 Increase PM-11 2 0.500 1.250 0.250 8 128 128   0.074 Increase PM-12 2 0.500 1.250 0.250 8 128 128   0.074 Increase Actinobacillus pleuropneumonia AP-1 0.5 0.125 0.313 0.063 2 128 4   0.081 Increase AP-2 0.5 0.125 0.313 0.063 2 128 4   0.081 Increase AP-3 0.5 0.125 0.313 0.063 2 128 4   0.081 Increase AP-4 0.5 0.125 0.313 0.063 2 128 4   0.081 Increase AP-5 One 0.250 0.625 0.125 8 128 32   0.040 Increase AP-6 One 0.250 0.625 0.125 8 128 32   0.040 Increase AP-7 One 0.250 0.625 0.125 8 128 32   0.040 Increase AP-8 One 0.250 0.625 0.125 8 128 32   0.040 Increase AP-9 One 0.250 0.625 0.125 8 128 32   0.040 Increase Mycoplasma hyopneumoniae MH-1 2 0.500 1.250 0.250 2 128 128   0.262 Increase MH-2 2 0.500 1.250 0.250 2 128 128   0.262 Increase MH-3 2 0.500 1.250 0.250 2 128 128   0.262 Increase MH-4 2 0.500 1.250 0.250 2 128 128   0.262 Increase MH-5 2 0.500 1.250 0.250 4 128 128   0.137 Increase MH-6 2 0.500 1.250 0.250 4 128 128   0.137 Increase MH-7 2 0.500 1.250 0.250 4 128 128   0.137 Increase Bodetella bronchiseptica BB-1 2 0.500 1.250 0.250 2 128 128   0.262 Increase BB-2 2 0.500 1.250 0.250 2 128 128   0.262 Increase BB-3 One 0.250 0.625 0.125 2 128 128   0.131 Increase BB-4 One 0.250 0.625 0.125 2 128 128   0.131 Increase BB-5 One 0.250 0.625 0.125 8 128 128   0.037 Increase BB-6 One 0.250 0.625 0.125 8 128 128   0.037 Increase BB-7 One 0.250 0.625 0.125 8 128 128   0.037 Increase BB-8 One 0.250 0.625 0.125 8 128 128   0.037 Increase BB-9 One 0.250 0.625 0.125 8 128 128   0.037 Increase Staphylococcus aureus ATCC29213 QC-2 2 0.500 1.250 0.250 One 64 128   0.521 Increase Escherichia coli ATCC 25922 QC-1l One 0.250 0.625 0.125 One 32 128  0.271 Increase

Experimental Example 2: Examination of therapeutic effects of swine digestive and respiratory infections

(1) Experiment selection and dynamics experiment

Compound antibacterial agent was prepared in an amount of 20 g of enlofloxacin (E) per 1 kg of the test product and 50 g of sulfamethoxazole (S) in an amount of 10 g of trimethoprim (T), and a test for treating digestive and respiratory infections was conducted. The overall disease outbreak of the farm was observed in the case of digestive diarrhea and respiratory symptoms. Microbiological tests were performed on pigs with respiratory symptoms by randomly screening these pigs for digestive symptoms (diarrhea). As a result, pathogenic microorganisms were detected simultaneously in the birch and feces, and pigs showing clinical symptoms of digestive and respiratory diseases were preferentially placed in the test group. The pigs that showed coughing (coughing), tremur (progression) and diarrhea (diarrhea) were divided into 4 groups (NC (T-1: 1 kg / feed 1 ton, T-2: 3 kg / feed 1 ton, T-3: 9 kg / feed 1 ton) and a healthy control (C) Respectively. T-1 and T-2 are recommended at the recommended maximum capacity (1 kg per ton of feed) and the recommended maximum capacity (3 kg per ton of feed) and T-3 (9 kg per ton of feed) . All were fed for 5 days. The pigs of T-3 showed a tendency to avoid feed intake for one day compared to T-1 and T-2, but soon adapted.

The results are shown in FIG. 5. As shown in FIG. 5, according to the experimental results, the composition administration of the example (E: S: T = 2: 5: 1) showed an increase in body weight as compared with the NC group T-2 and T-3 showed higher body weight gain than C group.

(2) Experimental effect of digestive tract infection disease

FIG. 6 is a graph showing the effect of the composition of Example (E: S: T = 2: 5: 1) at the recommended dose (1 kg and 3 kg / ton) and the triple dose (dose for safety test) The degree of diarrhea was observed by comparing NCs not administered.

These results show that T-2 and T-3, which were administered with the composition of Example (E: S: T = 2: 5: 1), markedly decreased from day 1 and day 2, It was able to confirm that it stopped at the day.

(3) Experimental effect of respiratory infectious disease treatment

FIG. 7 shows the results of examining respiratory symptoms such as cough in pigs in the control group (C), the untreated group (NC), the recommended dose group (T-1, T-2) and the triple- Cough symptoms were investigated. The results showed respiratory symptoms such as coughing in pigs of all groups before administration of the composition of Example (E: S: T = 2: 5: 1). However, after the administration of the composition of Example (E: S: T = 2: 5: 1), both the recommended dose and the 3-fold higher dose group showed significant respiratory improvement compared with the control group. T-2 and T-3 recovered rapidly without any significant difference, but T-1 showed a moderate recovery rate. In contrast, the untreated group (NC) remained symptomatic.

(4) Clinical outcome of gastrointestinal and respiratory infections

Figures 8 and 9 quantify statistical clinical indices by quantifying digestive (diarrhea) and respiratory (cough) symptoms, respectively. (E: S: T = 2: 5: 1) at the recommended doses T-1 and T-2 as feed additives, 1) was administered, and the clinical indices were observed. T-1, T-2, and T-3 began to disappear after administration of the composition and showed marked improvement in clinical symptoms compared to the group administered on day 7 and the group not administered on day 7.

Claims (4)

A composite antimicrobial composition for animals comprising 10-35% by weight of enlofloxacin, 5-25% by weight of trimetapririm and 40-80% by weight of sulfamethoxazole.
The complex antimicrobial composition according to claim 1, wherein enlofloxacin: trimethoprim: selfamethoxazole is mixed in a weight ratio of 2: 1: 5.
An animal drug for treating a gastrointestinal tract infection or a respiratory tract infection disease in an animal comprising the complex antimicrobial composition for animals of claim 1 or 2 as an active ingredient.
The veterinary pharmaceutical according to claim 3, which is formulated as a powder, pellet, granule, liquid, injection or ointment.

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