KR101118699B1 - Composition for treatment of Porcine proliferative enteropathy comprising chlorate - Google Patents

Abstract

The present invention relates to a composition for treating swine proliferative enteritis comprising chlorate and a method for treating swine proliferative enteritis by feeding the composition to pigs.

The composition of the present invention is safe for the living body, and can effectively treat swine proliferative enteritis.

Swine proliferative ileitis, chlorate, ume, sodium nitrate

Description

Composition for treatment of Porcine proliferative enteropathy comprising chlorate

The present invention relates to a composition for treating swine proliferative enteritis comprising chlorate and a method for treating swine proliferative enteritis by feeding the composition to pigs.

Porcine proliferative enteropathy (PPE) is a Gram-negative, curved rod type parasitic within the organizing cell that was identified as a pathogen by a British pathologist in 1994 and was named Lawsonia intracellularis ( L. intracellularis ) in the United States in 1997. As a bacterium, it is a disease known to cause diseases in horses, dogs, foxes, ferrets, primates, deer, ostriches, rats, hamsters, etc., in addition to pigs. In pigs, it spreads mainly through oral infection through feces. In particular, piglets are known to be infected after weaning through the papilla of contaminated sows in mammals.

L. intracellularis causes histological changes in the mesenteric area 8-10 days after infection, and clinical symptoms can be observed around 10-14 days, and a long incubation period when clinical symptoms peak at 21 days (16-17 weeks of age). Characterized in that, the feces are discharged to the body for several weeks through the feces.

Swine proliferative ileitis is one of the pig wasting diseases that causes enormous economic losses to pig farms by reducing feed intake and weight gain by causing thickening and inflammation of the ileal ends of growing pigs and slaughtered pigs. In Korea, 828 samples were collected from 65 farms nationwide in 2000, and all the farms surveyed were exposed to 100% pig proliferative colitis. Mari has been reported to have a positive rate of 56.4%.

Swine proliferative colitis is a disease that occurs mostly in pig farming countries around the world, including Europe, North America, Australia, New Zealand, South Africa, Brazil, Mexico, Japan, Taiwan, and Thailand. In the United States, the rates of farming and individual infections of swine proliferative colitis are known to be 96% and 37%, respectively, with significant economic losses of more than $ 10 million per year due to reduced feed efficiency, weight gain, and mortality. It is known.

In Korea, studies on swine proliferative colitis are partially conducted. However, the culture and characterization of L. intracellularis , the causative agent of proliferative colitis, is poorly performed. This has not been established.

Currently, the main treatment method for swine proliferative colitis is to add oxytetracycline (400ppm) or tylosin (100ppm) to the feed, and then re-administer the drug for 3-4 weeks after taking 2 weeks of dosing. It is true. The concentration of antibiotics used in the treatment of swine proliferative colitis is higher than two to three times the usual recommended therapeutic dose. Treatment of swine proliferative colitis, which requires long-term use of excess antibiotics, is not available for pigs in the rearing season due to the possibility of antibiotic residue in meat at the time of release.

Meanwhile, due to the global trend of regulating the use of antibiotics in livestock, there is an urgent need for the development of antibiotic replacement therapy and disease prevention program along with the prohibition of the use of some feed additive antibiotics. The development of therapeutic agents that can replace existing antibiotics (oxytetracycline, tylosin, tiamulin, etc.) is very early in the world. In addition, securing the market for swine proliferative colitis is a very important task.

Accordingly, the present inventors made efforts to develop a swine proliferative colitis treatment that can replace antibiotics, confirming that chlorate can safely and effectively treat swine proliferative colitis, and to complete the present invention. Reached.

One object of the present invention is to provide a composition for treating swine proliferative enteritis comprising chlorate.

Another object of the present invention is to provide a method for treating swine proliferative enteritis by feeding the composition to pigs.

"Chlorates" in the compositions of the invention are intended to be included at a concentration of 5 mM to 75 mM, preferably 5 mM to 45 mM, more preferably 15 mM. If the chlorate is 5 mM or less, the effect of treating swine proliferative colitis is insufficient (see Example 3), and if the chlorate is 75 mM or more, the kidney and liver are highly likely to cause toxicity (see Example 7). The chlorate salt may be sodium chlorate, potassium chlorate, magnesium chlorate, and sodium chlorate is particularly preferred.

The composition of the present invention may further comprise at least one or more of a mae extract and sodium nitrate together with chlorate.

"Oil extract" in the composition of the present invention is preferably to be included in a concentration of 0.05 to 1.0%, more preferably 0.1%. When the concentration of the ume extract is 0.05% or less, the effect of treating porcine proliferative colitis is less, and even when the concentration of the extract is greater than 1.0%, the intake decreases due to the astringent taste of the ume, so that the effect of treating porcine proliferative colitis is reduced (see Example 2). However, if an excipient or diluent is added to the composition of the present invention to alleviate the astringent taste of the mae extract, the mae extract may be included in a concentration of 1.0% or more.

"Omae" is an unripe blue plum of Prunus mune Sieb. Et Zucc. Of Rosaceae, which has been burned at low temperatures around 40 ° C, dried or smoked, and turned black. Is known for its warm nature. The constituents of OHMA are volatile components, flavonoids, cytosterol and organic acids, which include citric acid, malic acid, succinic acid, and tartaric acid. The reported pharmacological effects of five medium were antibacterial activity against anthrax, diphtheria, staphylococci, Bacillus and pneumococci, and hexane X showed antimutagenic activity and 5% maehwan significantly reduced the activity of roundworm. Known.

"Oil extract" of the present invention can be obtained by pouring water on the ume and applying heat to separate the supernatant and then filtered.

"Sodium nitrate" in the composition of the present invention is not particularly limited, but may be included in a concentration of 1 to 5mM, preferably 2.5mM.

The composition of the present invention is preferably formulated in the form of drinking water so that the pigs can freely ingest it.

The composition of the present invention may further comprise suitable excipients and diluents commonly used in the manufacture of veterinary compositions.

Excipients and diluents that may be included in the veterinary composition of the present invention include lactose, textose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulite Los, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, cetanol, stearyl alcohol, liquid paraffin, sorbitan monostea Latex, polysorbate 60, methylparaben, propylparaben and mineral oil.

The composition of the present invention may further include fillers, anti-coagulants, lubricants, wetting agents, spices, emulsifiers, preservatives and the like. Veterinary compositions according to the invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to the animal. The formulations may be in the form of powders, granules, tablets, capsules, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, suppositories, sterile injectable solutions, sterile external preparations and the like. The veterinary composition of the present invention so formulated may be administered via several routes including oral, transdermal, subcutaneous, vein, dural or muscle.

The composition of the present invention may vary depending on the age, sex, and weight of the animal, but the amount of 0.1 to 100 mg / kg may be administered once to several times a day. In addition, the dosage may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. The dosage does not limit the scope of the invention in any aspect.

The administration period of the composition of the present invention is also not particularly limited, but 1 to 30 days are appropriate, preferably 3 to 10 days, most preferably 7 days.

The composition of the present invention is safe for living body in a certain concentration range, and can effectively treat swine proliferative enteritis.

In addition, the composition of the present invention can be used as an antibiotic replacement, and can solve various problems due to the use of antibiotics because it is a combination of minerals and natural products.

In addition, the composition of the present invention can prevent and overcome the problem that the feed efficiency is lowered and the growth and growth is delayed due to swine proliferative enteritis.

Hereinafter, the examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, those skilled in the art. Will be self-evident.

Example 1 Extraction of Hot Water and Drying

The hot water extraction of ume was poured into 100 g of ume in water in a shaker to make 1000 mL, heated for 3 hours, and then the supernatant was separated and filtered after vacuum reduction. In order to confirm the recovery rate of the hot water extract was compared using two methods. In the first method, the filtered X component was evaporated at 60 ° C. using a rotary vacuum evaporator, and then completely dried to dry powder in a dry oven. In the second method, the filtered X component is evaporated at 60 ° C. using a rotary vacuum evaporator, and then decompressed in a hot water bath. Concentrate and freeze with freeze dryer. Dried to powder. The recoveries of these powders were 4.6% when dried in a dry oven and 4.5% when dried in a freeze dried oven. The recovery rate was slightly higher when dried in a dry oven, but a large difference was found between the two methods. Was not. In this example, the freezing? The powder of the hot plum extract was prepared by the drying method, and the prepared powders were stored in the freezer and used in the later examples.

Example 2 Determination of Application Concentration Ranges of O medium

2-1. Experiment method

Experimental Animals: Male ICR mice (4 weeks old) were used in the experiments, and the rearing environment was 23 ± 3 ℃, 50 ± 10% relative humidity, and 12 hours light cycle (lighting at 7:00, light at 19:00), Roughness was adjusted to 150-300 Lx to maintain a constant breeding environment conditions. In addition, feed and water can be freely consumed.

Infection of L. intracellularis: L. intracellularis in order to infect mice, and after separating the pig mucosal hyperplasia of the president sites infected with L. intracellularis crusade was collected from pigs infected with Vibrio suspended in saline, using a catheter 1 ml of mice were orally administered to infect L. intracellularis .

Experimental design: Twenty five infected mice (average weight: 29.4 ± 3.3g) whose feces were collected from the L. intracellularis were detected by PCR, and the concentrations (0, 0.05, 0.1, 0.5, 1.0%) and five mice in one group were administered to the control group and four mae extract extracts for 5 days as drinking water in a free intake mode, and observed diarrhea reduction over time. The effect was compared and examined.

2-2. Experiment result

In mice infected with L. intracellularis , the main symptom was diarrhea or stool, as in pigs infected with proliferative colitis, and hemorrhagic diarrhea in severe cases. Therefore, the rats infected with Lawsonia were observed fecal state changes after ingesting the ume extract by concentration. Table 1 shows the grade according to the fecal state. Normal feces were set to 0 and watery diarrhea was 3 to analyze the observed fecal state.

Diarrhea class according to fecal status Diarrhea grade Fecal state 0 normal One Weak diarrhea 2 diarrhea 3 Watery diarrhea

Table 2 shows the analysis by scoring the stool status according to the grade after administering the mae extract by concentration. According to Table 2, all the groups administered the ume extract showed a result that gradually improved over time. The most effective branch office effect was observed in the MF-0.1 group administered with 0.1% of the mae extract among the group administered the mae extract. The MF-0.05 group administered with 0.05% omae extract showed some effects over time, but it did not show any significant effect compared to the control group. In addition, in the MF-0.5 and MF-1.0 groups administered at high concentrations of 0.5 and 1.0%, the branch effect was very low over time, especially in the MF-1.0 group with the highest concentration. However, no significant difference was observed in comparison with the control group.

Therefore, from these results, the concentration of the most effective ume hot water extract to 0.1% to alleviate the symptoms of diarrhea and bloody stool due to swine proliferative enteritis, and through the complementary action with other drug components, swine proliferative colitis It was applied to show the effect in the treatment.

Example 3: Test of Concentration Range of Sodium Chlorate

3-1. Experiment method

Experimental animals: ICR mouse males were used for 4 weeks of age, and the breeding environment was 23 ± 3 ℃, relative humidity 50 ± 10%, lighting for 12 hours (cycled to 7:00, lights out 19:00), and illumination was Adjusted to 150-300 Lx to maintain a constant breeding environment conditions. In addition, feed and water can be freely consumed.

Infection of L. intracellularis: L. intracellularis in order to infect mice, and after separating the pig mucosal hyperplasia of the president sites infected with L. intracellularis crusade was collected from pigs infected with Vibrio suspended in saline, using a catheter 1 ml of mice were orally administered to infect L. intracellularis .

PCR Analysis: Prior to performing the PCR analysis, agarose gels must first be made. 1 g of agarose was dissolved in 100 ml of TAE buffer to prepare a 1% agarose gel. For 50-fold TAE buffer solution, 242 g (2M) tris (Trizma base), 57.1 ml (1M) glacial acetic acid (100% acetic acid), 100 ml 0.5 M Na ₂ -EDTA (pH 8.0), etc. To 1L to prepare. Agarose was quantified using a chemical balance, placed in a flask, filled with a buffer solution, and dissolved by heating slightly in a microwave oven. The gel was poured into the gel mold before the agarose hardened and left to stand for about 15 minutes to harden.

The electrophoretic apparatus was properly filled with TAE buffer, the previously prepared agarose gel was in place, and 5 μl PCR product was placed in the well of the gel. About 3 μl of the marker was added to another well, and then run at 50 V for about 50 minutes. Then, a band formed on the agarose gel was identified using a UV transilluminator (Bio-Rad) at 254 nm.

Experimental design: Forty infected mice (average body weight, 27.6 ± 2.9 g) whose feces were collected from the L. intracellularis were identified by PCR, and each group was divided into 10 rats. Drug group), dosing group I (5 mM chlorate + 2.5 mM sodium nitrate + 0.1% berry extract (HFWE)), dosing group II (15 mM chlorate + 2.5 mM sodium nitrate + 0.1% HFWE), and dosing group III (45 mM Chlorate + 2.5 mM sodium nitrate + 0.1% HFWE), designed to be freely ingested through drinking water. In addition, feed was freely consumed during the experiment.

Fecal samples were taken daily for each group over time and analyzed for the presence of L. intracellularis .

3-2. Experiment result

ICR mice infected with L. intracellularis , the causative agent of swine proliferative ileitis , were administered via drinking water, and PCR analysis was performed on feces of mice to confirm the effects of L. intracellularis on drugs over time. Was performed. In order to perform PCR amplification, a request was made to Bioneer Co., Ltd. to prepare a primer. Prepared primers were named LI-2F, LI2-R (about 319bp). Each nucleotide sequence is LI-2F; 5'-TATGGCTCTCAAACACTCCG-3 '(SEQ ID NO: 1), LI-2R; 5'-TGAACCTATTCCTATTCTCC-3 '(SEQ ID NO: 2) and the PCR program was performed at 95 ℃-30 seconds, 56 ℃-30 seconds, 72 ℃-60 seconds, 45 conditions. The PCR mixture was reacted with 5 μl template DNA, 10 μl of PCR premixture (Intron, Korea), 1 μl of each primer (100 pM), and 3 μl of DW to a final volume of 20 μl.

Table 3 shows the results of investigating the number of infected mice over time in the control group (no-dose group) and the dosing group (I-III) of mice infected with L. intracellularis by PCR. In case of dosing group I, there was a slight therapeutic effect over time, and the treatment rate was 20% at 7 days. In the case of the dosage group II, the dose-response relationship was relatively well shown according to the dosage, and all the results were completely cured on the 7th day of the administration. On the other hand, in the case of dosing group III, the tendency was similar to that of dosing group I, and the treatment rate was 20% on the 7th day of dosing.

Therefore, the preliminary efficacy test, the field application test, and the safety test for pigs were performed using the composition of the dosage group II (15 mM chlorate + 2.5 mM sodium nitrate + 0.1% HFWE) and the administration period. Based on these results, the final dose of the safe and effective drug was determined.

Example 4: Screening Test for Pigs of Compositional Therapeutics

4-1. Experiment method

Experimental animals: 10 weeks of age having diarrhea (including Yanbian) at Senon test farm located in Hamyang-gun, Gyeongsangnam-do? Fecal samples were collected from the rearing pigs, and experiments were performed on 15 pigs (31.4 ± 1.2 kg) that were confirmed to be infected with swine proliferative colitis by PCR.

Experimental design and dosing: Each experimental group consisted of five initial pigs, and the dosing group consisted of dosing group I (5 mM chlorate + 2.5 mM sodium nitrate + 0.1% MFWE) and dosing group II (15 mM chlorate + 2.5 mM sodium nitrate + 0.1). % MFWE), and the control group (non-dose group) designed to perform the experiment.

The drug was dissolved in drinking water and consumed freely, and the feed was freely consumed.

Sampling and Analysis: The feces of all pigs used in the experiments were collected daily using a cotton swab over time while administering a drug prepared through drinking water. In order to purify the DNA of L. intracellularis from the collected feces, 0.2 g of the resulting fecal was purified by 950 μl of lysis buffer (5 M guanidine thiocyanate, 22 mM EDTA, 0.05 M Tris-Cl; pH6.4, and 0.65% Triton X-). 100), reacted at room temperature for 60 minutes, centrifuged at 14,000 xg for 20 seconds, and mixed the supernatant with 50 µl of diatomaceous earth suspension (20% diatomaceous earth, 0.17 M HCl) for 10 minutes at room temperature. After the reaction, the supernatant was again removed by centrifugation (14,000 xg, 20 sec), and the pellet was dried at 56 ° C and suspended in 20 µl TAE buffer solution (Tris; 10 mM, EDTA; 1 mM, pH 0.8). . After suspension, 14,000 xg, centrifugation for 20 minutes, the supernatant was used for the test.

After completion of the PCR reaction, the samples were electrophoresed on an agarose gel containing ethidium bromide at a final concentration of 0.5 µg / µl and identified under a UV transilluminator (Bio-Rad) at 254 nm.

4-2. Experiment result

Pigs infected with swine proliferative colitis show stool or diarrhea and, in severe cases, hemorrhagic diarrhea. As a result, feed efficiency is reduced and growth and growth are delayed. The ileum of pigs infected with swine proliferative colitis is apparently bleeding, and the barrier is thickened.

Table 4 shows the results obtained by analyzing the PCR method by collecting the stool with drinking water after administration of the drug in the growing pigs infected with swine proliferative ileitis in drinking water. From the results of this experiment, no change was found in the control group without any treatment, whereas in the administration group I, the effect of the medication administration was gradually occurring compared to the administration group II, but from the third day of the medication administration On the 5th day of treatment, the treatment rate was 40% and on the 7th day, the treatment rate was 60%. These results showed that the treatment rate of L. intracellularis in mice was slightly higher than that of the treatment group I, compared to 20% at 7 days. On the other hand, in the case of dosing group II, the treatment effect for swine proliferative colitis was started from the 2nd day of the administration, and the treatment rate was 60% on the 3rd day of the administration and 80% on the 5th day of the administration. On the 7th day of the last day of administration, all of the pig proliferative colitis were treated. The degree of severity of the disease varies according to the susceptibility of the pigs infected with swine proliferative colitis. In addition, this is consistent with the results of the treatment of L. intracellularis using the mouse, which was completely cured on the 7th day of the administration of the group II drug.

From the above results, it was confirmed that the drug consisting of 15mM chlorate, 2.5mM sodium nitrate and 0.1% ume hydrothermal extract (MFWE) was effective in treating swine proliferative colitis, and it was appropriate to administer for 7 days through drinking water. appear. Thereafter, the field application test confirmed the therapeutic effect of the drug against swine proliferative colitis.

Example 5 Field Application Test of Porcine Proliferative Colitis

5-1: Experimental Method

METHODS: To determine the therapeutic effect of a treatment for swine proliferative colitis, 258 pigs at around 10 weeks of age in three pig farms in Hamyang, Jinju, and Gimhae in Gyeongsangnam-do were reviewed. Infected pigs should be selected before 10 weeks of age with diarrhea (including Yanbian) in each pig farm. Fecal samples were collected from the pigs, and then confirmed by the PCR technique.

Experimental Design and Dosing: The experiments were carried out by selecting 10-15 pigs in a single room by selecting infected pigs for each farm. Drug administration was designed to be administered for 7 days by mixing freely the drug (15mM chlorate + 2.5mM sodium nitrate + 0.1% MFWE) through drinking water for all experimental pigs. In addition, the feed was also allowed to eat freely without limited diet.

Sampling and Analysis: All pigs used in the test for each time of administration using the drug prepared through drinking water were collected for 1, 3, 5, and 7 days after dosing with a cotton swab. The presence of L. intracellularis bacteria was confirmed using.

5-2. Experiment result

Figure 1 shows the results of the PCR test on the feces after administration of the drug to pigs infected with proliferative colitis. (A), (B) and (C) show the results of diagnosis of L. intracellularis by PCR for fecal samples on the 1st, 3rd and 7th day of dosing, respectively. Positive samples of 9 and 10 lanes (swine infected with proliferative colitis) showed complete cure on day 7 of dosing.

Administration of the drug to pigs infected with proliferative ileitis resulted in 5 (17.4%) after 1 day of administration, 115 (44.6%) after 3 days of administration, 76 (29.5%) after 5 days of administration, and last dose 7 After 16 days (6.2%), the treatment efficiency was 97.7%, which is 252 out of 258, and 6 (2.3%) were not effective. In all six cases, which did not show a therapeutic effect, the drinking water and feed intake were not properly performed due to severe respiratory diseases.

Table 5 shows the results of field tests administered for 7 days with treatment water prepared for 258 pigs infected with swine proliferative colitis.

Example 6 Safety Test of Porcine Proliferative Colitis Therapeutic (Oral Toxicity Test of Compositional Treatment Using Pig)

6-1. Experiment method

Experimental Animals: 22 healthy three-way hybrid (Landrace x Yorkshire x Duroc) male pigs before and after 10 weeks of age were purchased from a farm near Jinju, Gyeongsangnam-do for 7 days of short-term toxicity testing. The general symptoms were observed for one week, and only healthy pigs without any abnormalities were selected and used for the test. The weight of the test was 31.3 ± 0.31kg.

The conditions of the breeding environment were maintained at the temperature of 20 ± 1.5 ℃ and the relative humidity of 60 ± 10% until the end of the test. Feed (Samyang Feed, Seoul) was supplied for free intake, and the composition was supplied to drink freely by mixing in drinking water by varying the concentration of sodium chlorate.

Setting of Dosage and Test Substance Administration: The test group was composed of weighing pigs before and after 10 weeks of age determined to be healthy during the acclimation period and assigning 5 animals per group by random method according to the weight range.

The dose was set to 3, 5, and 10 multiples for the concentration of chlorate (15 mM) among the components that had a therapeutic effect on swine proliferative colitis. Thus, control group (untreated group), dose group I (3 x 15 mM chlorate + 2.5 mM sodium nitrate + 0.1% MFWE), dose group II (5 x 15 mM chlorate + 2.5 mM sodium nitrate + 0.1% MFWE), and dose group III (10 × 15 mM chlorate + 2.5 mM sodium nitrate + 0.1% MFWE).

Preparation and Administration of Test Substances: The prepared therapeutic agents with different concentrations of chlorate were dissolved in water, stirred well, and then dissolved in drinking water.

Observation and Inspection Items:

Clinical symptoms observation-General symptoms and deaths were observed once daily during the study period.

Body weight-The body weight of the test animals was measured at the start of the test and the end of the test.

Autopsy Findings-After the end of the study, all animals were bled from the jugular vein before anesthesia and the autopsy was performed to observe organs visually.

Hematological examination-RBC, WBC, PCV, and hemoglobin (Hb) were measured from blood drawn through the jugular vein. Some blood was allowed to stand at room temperature for 30 minutes, and then serum was separated by centrifugation at 3,000 rpm for 15 minutes, and aspartate aminotransferase (AST), alanine aminotrasferase (ALT), blood urea nitrogen (BUN), and creatinine were autoanalyzer. (CL-7200, Shimadzu Co., Japan).

Histopathological examination-After fixation of the liver and kidneys obtained by autopsy in 10% formalin solution, paraffin sections were prepared and hematoxylin-eosin staining was performed as in the usual method. The process was observed by light microscopy.

Statistical analysis: Statistical analysis of the results was performed by student's t-test using Sigma plot, and statistically significant difference was found when p <0.05.

7-2. Experiment result

General Symptoms Observed: 7 days after the administration of 3, 5, and 10 times the concentration of chlorate (15 mM) in drinking water in the composition of a therapeutic agent that was effective for treating proliferative colitis in pigs. This was not observed and no animals died.

Body weight change: Table 6 shows the body weights at the start of the test and at the end of the test 7 days in the control and drug administration groups. In the case of dosing group I, the body weight at the end of the test did not show a difference compared to the control group, and in the case of the dosing group II, the average weight was slightly decreased compared with the control group, but there was no statistically significant difference. In the case of the dosage group III, the weight loss was found to be slightly larger than the control group, but there was no statistically significant difference. Therefore, no significant difference was found in body weight change at the end of the test in both control and experimental groups.

Autopsy Findings: Since no animals died during the test period, all animals were autopsied to determine gross findings. In the case of dosing group I and II, no abnormalities were found in all animals, but in the case of dosing group III, liver size was confirmed in some animals.

Hematological tests: Table 7 shows the concentrations of the toxic chlorate in the components of the pig propagatory colitis treated for 7 days at 3 x 15, 5 x 15, and 10 x 15 mM, respectively. Blood samples are collected and analyzed for changes in hematological and biochemical markers in the blood. Looking at Table 7, in the case of the dosing group I, compared to the control group, all the values of the indicators were slightly changed, but it was confirmed that all were within the range of normal values.

In case of dosing group II, the values of RBC, WBC, PCV, and Hb are all slightly out of the normal range, and the experimental animals showing the values within the normal value range and the animals out of the value are mixed. . In addition, the ALT and AST values are within the normal range, but in this case, the experimental animals exhibiting the values within the normal range and the experimental animals out of the values are mixed. The values of BUN and creatinine were lower than the range of normal values, which also showed a little more mixture of experimental animals outside the normal values than those of the normal values.

For dosing group III, all indicators are outside the normal range except for the RBC value. These results indicate that there is a high possibility of causing toxicity in experimental animals in general, such as liver and kidney. In other words, AST and ALT, which are indicators of the effects on liver function as liver enzymes, and BUN and creatinine values, which are related to renal function, are all highly likely to cause liver and kidney disorders.

Histopathological observation: Figure 2 shows the pathology of kidney tissue for each group after administration of drinking water for 7 days at different concentrations of chlorate in the components of the therapeutic drug to confirm the toxicity of the antibiotic-substituted swine proliferative colitis. This is a sample of tissue prepared for the purpose of scientific observation.

Figure 2A shows the kidney tissue of the control pigs administered nothing, B shows the kidney tissues of pigs administered with a combination of 3 x 15mM chlorate, 2.5mM sodium nitrate and 0.1% MFWE for 7 days, Overall, clean and healthy, like the structure of normal tissue was observed. C represents the kidney tissue of pigs treated with a combination of 5 x 15 mM chlorate, 2.5 mM sodium nitrate, and 0.1% MFWE for 7 days in drinking water. Partial necrosis of the tubules and expansion of the tubules were observed. D represents the kidney tissue of pigs treated with a combination of 10 x 15 mM chlorate, 2.5 mM sodium nitrate, and 0.1% MFWE. Degeneration was observed.

Figure 3 is administered for 7 days with drinking water at different concentrations of chlorate in the components of the therapeutic agent to confirm the toxicity of the antibiotic replacement pig proliferative colitis, the purpose of histopathological observation of the liver tissue for each group It shows the fabricated sample.

FIG. 3A shows liver tissue of control pigs administered nothing, B represents liver tissue of pigs administered with a combination of 3 × 15 mM chlorate, 2.5 mM sodium nitrate and 0.1% MFWE for 7 days, Overall, a healthy appearance was observed, similar to the structure of normal tissues. C represents the liver tissue of pigs administered with a combination of 5 x 15 mM chlorate, 2.5 mM sodium nitrate and 0.1% MFWE for 7 days in drinking water, necrosis of some tissues, deficiency and collapse of cell nuclei were observed. And D represents the liver tissue of pigs that received a combination of 10 x 15 mM chlorate, 2.5 mM sodium nitrate and 0.1% MFWE through drinking water for 7 days, with C more severe tissue necrosis, cell nucleus deficiency and collapse. Etc. were observed.

1 shows the results of diagnosing PPE using PCR in pig feces, (A) on day 1, (B) on day 3, and (C) on day 7

Figure 2 shows a tissue sample of kidney tissue of pigs administered various concentrations of chlorate for 4 weeks through saline.

Figure 3 shows a tissue sample of liver tissue of pigs administered various concentrations of chlorate for 4 weeks through saline.

<110> INDUSTRY-ACADEMIC COOPERATION FOUNDATION GYEONGSANG NATIONAL UNIVERSITY <120> Composition for treatment of Porcine proliferative enteropathy          comprising chlorate <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 1 tatggctctc aaacactccg 20 <210> 2 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 2 tgaacctatt cctattctcc 20  

Claims (7)

A pharmaceutical composition for treating swine proliferative ileitis, comprising 15 mM sodium chlorate, 2.5 mM sodium nitrate, and 0.1 wt% phenolic hot water extract. delete delete delete delete delete The pharmaceutical composition of claim 1, wherein the composition is formulated in one or more dosage forms for oral administration selected from the group consisting of powders, granules, tablets and capsules.

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