Veterinary compositions for alleviating or inhibiting E. coliosis in poultry and their administration methods

Abstract

The veterinary composition for alleviating or inhibiting E. coli in poultry, which is an embodiment, comprises florfenicol, acetaminophen, and bromhexine as active ingredients. The method of administration of the composition of the embodiment is applicable as a liquid oral form. The embodiment exhibits excellent stability and efficacy as a combination product, and in particular, the stability of the formulation can be maintained for a long period, so it can be efficiently utilized for alleviating or inhibiting E. coli in poultry.

Description

Technology Field

[0001] An embodiment relates to a veterinary composition for alleviating or inhibiting E. coli infection in poultry and a method of administering the same. Background Technology

[0003] Escherichia coli exists as part of the normal intestinal flora of birds and mammals. It causes various diseases by forming lesions in the intestines or digestive tract, either directly or through secondary infections, or by co-infecting with other bacteria and viruses. In chickens, symptoms manifest as various forms, including sepsis, air sacsitis, preophthalmitis, arthritis, salpingitis, and E. coli granulomatosis. Infections caused by these pathogens often occur through poultry products and develop into serious public health issues.

[0004] Meanwhile, florfenicol is a thiamphenicol derivative successfully synthesized by Schering-Flow in the United States in 1979. This antibiotic has a broad antibacterial spectrum and is useful even against strains resistant to thiamphenicol and chloramphenicol.

[0005] Related prior art includes Korean registered patent 10-1309583, a feed additive composition for preventing infectious diseases in chickens, Korean registered patent 10-2125235, a novel bacteriophage for preventing and treating E. coli in chickens and an antimicrobial composition using the same. The problem to be solved

[0007] The purpose of the embodiment is to provide a veterinary composition for alleviating or inhibiting E. coli infection in poultry and a method for administering the same. means of solving the problem

[0009] To achieve the above objective, a veterinary composition for alleviating or inhibiting E. coli infection in poultry according to one or more embodiments comprises florfenicol, acetaminophen, and bromhexine as active ingredients.

[0010] The above composition may be a liquid oral administration composition.

[0011] The above composition may further include any one additive selected from the group consisting of dimethylacetamide, methylpyrrolidone, dimethylformamide, and combinations thereof.

[0012] The above composition may further include any one excipient selected from the group consisting of propylene glycol, polyethylene glycol, glycerin, and combinations thereof.

[0013] The above composition may contain 10 to 30 parts by weight of florfenicol based on 1 part by weight of bromhexine.

[0014] The above composition may contain 10 to 30 parts by weight of acetaminophen based on 1 part by weight of bromhexine.

[0015] The above composition can reduce the number of E. coli in poultry after 3 days of administration.

[0016] The above composition may include 0.7 to 1.3 parts by weight of acetaminophen based on 1 part by weight of florfenicol.

[0017] To achieve the above objective, a method of administering a veterinary composition for alleviating or inhibiting E. coli in poultry according to one or more embodiments involves mixing the veterinary composition for alleviating or inhibiting E. coli in poultry described above with drinking water and administering it orally in an amount of 0.5 to 2 mg of the bromhexine per 1 kg of body weight of the poultry individual.

[0018] With the above administration method, compared to poultry infected with E. coli, poultry infected with E. coli after the start of administration may have increased weight gain and improved feed requirements.

[0019] The above administration method may have antipyretic and expectorant effects on poultry with E. coli infection. Effects of the invention

[0021] The veterinary composition for alleviating or inhibiting E. coli in poultry and the method of administration thereof according to the embodiment are applicable as a liquid oral formulation and have excellent stability and efficacy as a combination product, and in particular, the stability of the formulation can be maintained for a long period, so they can be efficiently utilized for alleviating or inhibiting E. coli in poultry. Specific details for implementing the invention

[0023] Hereinafter, embodiments of the present invention are described in detail so that those skilled in the art can easily implement the invention. However, the present invention may be embodied in various different forms and is not limited to the embodiments described herein. Throughout the specification, similar parts are denoted by the same reference numerals.

[0024] The terms “pharmacologically (or veterinarily) acceptable salts thereof” or “salts thereof” described herein in relation to a specific compound mean salts that retain the biological efficacy and properties of the parent compounds and are not biologically or otherwise harmful when a single dose is administered. For example, “pharmacologically (veterinarily) acceptable salts thereof” or “salts thereof” may be base-added salts of the specific compound and may be prepared from inorganic and organic bases. Salts derived from inorganic bases may include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, and magnesium salts. Salts derived from organic bases may include, but are not limited to, primary, secondary, and tertiary amines; substituted amines, including naturally occurring substituted amines; and may include salts of cyclic amines including isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucarmine, theobromine, purine, piperazine, piperidine, and / or N-ethylpiperidine. Additionally, carboxylic acid amides including other carboxylic acid derivatives, such as carboxamides, lower alkyl carboxamides, di(lower alkyl)carboxamides, etc., may also be useful in the practice of the present invention.

[0025] As used herein, the terms "prevention" or "mitigation" refer to the act of inhibiting the progression, inhibiting the occurrence, or mitigating the severity of E. coli infections and related symptoms in poultry through the administration of the preparation of the present invention.

[0026] As used herein, the term "treatment" refers to the act of improving or beneficially altering symptoms of E. coli infection in poultry or related symptoms such as fever and expectoration by administering the preparation of the present invention, and refers to the act of eliminating clinical symptoms or alleviating the severity of their occurrence. For example, the "treatment" may include the alleviation or improvement of one or more symptoms or conditions among symptoms of E. coli infection and symptoms of medication stress, reduction of the range of symptoms, stabilization of symptoms, non-spread of symptoms, suppression of symptom occurrence, delay or slowing of symptom progression, improvement or alleviation of the symptom state, and partial or total reduction of symptom occurrence.

[0027] In describing the present invention in this specification, no distinction is made between the "singular" and "plural" of nouns, and singular expressions are used as a concept including both singular and plural, except where it is clear from the context that the singular refers only to the singular.

[0028] Unless otherwise specifically stated in this specification, "or" and "and / or" are used interchangeably, and the notation "A or B" or "A and / or B" may be interpreted to include the meaning of "A, B, or, A and B".

[0030] The following describes an example of implementation in more detail.

[0031] To achieve the above objective, a veterinary composition for alleviating or inhibiting E. coli infection in poultry according to one or more embodiments comprises florfenicol, acetaminophen, and bromhexine as active ingredients.

[0032] Colibacillosis in poultry (e.g., chickens) is caused by avian pathogenic Escherichia; E. coliThis disease, caused by infection with APEC, manifests various lesions such as airsacculitis, perihepatitis, peritonitis, pericarditis, salpingitis, omphalitis, and osteomyelitis, leading to stunted growth and economic losses due to mortality in poultry.

[0033] The main symptoms of E. coli infection commonly occur in poultry aged 6 to 10 weeks, and when E. coli is transferred to hatching eggs, the hatching rate decreases, and it can also occur in day-old chicks, causing umbilical cord infection, so the symptoms vary.

[0034] E. coli sepsis mainly occurs in poultry under 10 weeks of age, with mortality rates ranging from 5% to 50%, and chicks develop white or yellow-green diarrhea and swollen bellies.

[0035] When looking at diseases related to E. coli, first, E. coli is transferred to the eggs, causing a decrease in hatching rate, resulting in the occurrence of dead eggs, poor yolk absorption, and umbilical cord inflammation. The abdomen swells up, and the eggs die within 2 to 3 days after hatching.

[0036] Furthermore, if synovitis progresses chronically, the joints swell and the animal becomes limping. Salpingitis and peritonitis: If E. coli invades the air sacs within the abdominal cavity, air sac inflammation progresses to peritonitis and salpingitis, causing the fallopian tubes to swell and become inflamed, sometimes leading to death. Hemorrhage and ulcers may occur on the intestinal mucosa, and hemorrhagic enteritis may develop. E. coli granulomatosis is characterized by wart-like granulomatous lesions found in the small intestine, cecum, and liver.

[0037] The embodiment alleviates or suppresses the symptoms of E. coli infection in such poultry.

[0038] Florfenicol, the active ingredient, is included as an active ingredient in the embodiments, in the sense of including florfenicol or its salts. Florfenicol is a synthetic thiamphenicol derivative with a broad antibacterial spectrum and is veterinarily useful as a disinfectant and antimicrobial agent for animals. However, since it is not a formulation that dissolves easily in water, stability is an issue in aqueous formulations.

[0039] Acetaminophen, the active ingredient, is included as an active ingredient in the embodiment in the sense that it includes acetaminophen or a salt thereof. It has antipyretic and analgesic effects.

[0040] The active ingredient, bromhexine, is a synthetic derivative of the plant-derived active ingredient vasicine and exists in the form of bromhexine salts (e.g., bromhexine hydrochloride). Bromhexine is known to induce secretolytic and secretion-promoting effects in the bronchial region. It has a characteristic bitter taste.

[0041] The embodiment combines the three active ingredients mentioned above to effectively alleviate or suppress E. coli infection in poultry, and can induce an increase in weight gain or feed intake despite the infection. In addition, it can reduce hematological indicators such as WBC, MCH, and MCHC, reduce blood biochemical indicators such as ALT, AST, and unic acid creatine, and increase ALP. Furthermore, it can achieve the effects of reducing airway secretions and lowering body temperature.

[0042] The above composition may contain 10 to 30 parts by weight or 15 to 25 parts by weight of florfenicol based on 1 part by weight of bromhexine. In this case, an expectorant effect along with an antibacterial effect suitable for poultry E. coli infection can be obtained.

[0043] The above composition may contain 10 to 30 parts by weight or 15 to 25 parts by weight of acetaminophen based on 1 part by weight of bromhexine. In this case, antipyretic and analgesic effects suitable for poultry E. coli infection and expectorant effects can be obtained together.

[0044] The above composition may contain 0.7 to 1.3 parts by weight or 0.8 to 1.2 parts by weight of acetaminophen based on 1 part by weight of florfenicol. When florfenicol and acetaminophen are combined and applied within these ranges, the inhibition of E. coli and antipyretic effects can be efficiently obtained, and use within these ranges is considered to be a unique characteristic applicable to poultry, unlike large animals (e.g., pigs).

[0045] The above composition may be a liquid composition.

[0046] The above composition may be a liquid oral administration composition.

[0047] When the above composition is applied for liquid oral administration to poultry, it can be easily administered by mixing with drinking water, and relatively easily symptom relief or suppression effects can be obtained regarding the onset of E. coli infection.

[0048] Florfenicol, among the active ingredients, is known to be difficult to formulate into an aqueous form, and even if formulated into a liquid form, the stability of the liquid composition is significantly reduced, but the composition of the embodiment was able to solve this problem.

[0049] The composition of the embodiment may further include any one additive selected from the group consisting of dimethylacetamide, methylpyrrolidone, dimethylformamide, and combinations thereof.

[0050] The above additive helps in the liquid formulation of the active ingredient and can particularly help in the liquid formulation of florfenicol.

[0051] The above additive may be included in an amount of 1 to 3 parts by weight or 1.5 to 2.5 parts by weight based on 1 part by weight of florfenicol. In this case, it is advantageous for the liquefaction of the florfenicol-containing composition.

[0052] The composition of the embodiment may further include any one excipient selected from the group consisting of propylene glycol, polyethylene glycol, glycerin, and combinations thereof. The excipient may help stabilize the formulation of the florfenicol-containing composition.

[0053] The above composition may contain 0.3 to 2 parts by weight or 0.5 to 1.5 parts by weight of the excipient based on 1 part by weight of florfenicol. In this case, it is advantageous for stabilizing the florfenicol-containing liquid composition.

[0054] It was confirmed that the composition of the embodiment ensures the stability of the formulation for more than 8 months under harsh conditions (40°C and 75% relative humidity), and it is expected that the stability of the formulation will be ensured for more than 24 months at room temperature, which is the recommended storage condition.

[0055] The above composition can be mixed into the drinking water of poultry and administered orally. A specific method of administration will be described later.

[0056] The above composition can be prepared by mixing in the manner of preparing a conventional liquid veterinary composition, so a detailed description is omitted.

[0057] The above composition can induce excellent antipyretic and expectorant effects in poultry with E. coli infection.

[0058] The above composition can induce a reduction in the number of E. coli in poultry three days after the start of administration.

[0059] The above composition can induce an increase in weight gain in poultry infected with E. coli after the start of administration compared to poultry infected with E. coli.

[0060] Compared to poultry infected with E. coli, the feed requirement of poultry infected with E. coli after the initiation of administration of the above composition can be improved.

[0062] A method of administering a veterinary composition for alleviating or inhibiting E. coli infection in poultry according to an embodiment may be to mix the composition described above with drinking water and administer it orally in an amount of 0.5 to 2 mg of the bromhexine per 1 kg of body weight of the poultry individual.

[0063] A method of administering a veterinary composition for alleviating or inhibiting E. coli infection in poultry according to an embodiment may be to orally administer the composition described above by diluting it with drinking water such that the amount of bromhexine is 5 to 10 g per ton of drinking water.

[0064] Oral administration is recommended once a day for 5 days, but is not limited to this.

[0065] The above poultry may be chickens, and the effect is particularly excellent when applied to chickens (including chicks).

[0066] The above administration method can provide excellent antipyretic and expectorant effects for poultry with E. coli infection.

[0067] The above administration method can reduce the number of E. coli in poultry after 3 days of administration.

[0068] With the above administration method, the weight gain of poultry infected with E. coli after the start of administration may increase compared to poultry infected with E. coli.

[0069] With the above administration method, the feed requirements of poultry infected with E. coli can be improved after the start of administration compared to poultry infected with E. coli.

[0071] The following provides a more detailed explanation through specific embodiments. The following embodiments are merely examples to aid in understanding the present invention and do not limit the scope of the invention.

[0073] 1. Formulation and evaluation of formulation stability

[0074] A liquid composition sample was prepared with the composition as presented in Table 1 below.

[0076] The formulation stability evaluation confirmed whether the composition was stably mixed after manufacturing through appearance verification, pH testing, content testing, and microbial limit testing.

[0077] For the stability evaluation under harsh conditions, a constant temperature and humidity chamber was applied at 40°C and 75% relative humidity. Samples were taken at 2 weeks, 1 month, 2 months, 3 months, 4 months, 6 months, and 8 months of storage, respectively, starting from Day 0 (Initial). Three principal component analyses were performed for each period, evaluated based on the Initial (Day 0) date, and stability was confirmed up to the 8th month. O indicated that the active ingredient remained stable for more than 6 months under harsh conditions, while X indicated that some components decreased.

[0078] Content: parts by weight Composition 1 Composition 2 Composition 3 Composition 4 Composition 5 Composition 6 Florfenicol 20 20 20 20 20 20 Acetaminophen 20 20 - - - - Bromhexin 1 1 - - - - aspirin - - 20 20 20 20 Vitamin C - - 3 - 3 - DMAC (Dimethylacetamide) 50 - - - 47 50 NMP (N-methyl-2-pyrrolidone) - 50 47 50 - - ethanol - - - - QS QS potassium hydroxide - - - - QS QS surfactants - - 10 10 - - PG (propylene glycol) 15 15 - - - - Post-manufacturing stability evaluation o o - - - - Stability evaluation under harsh conditions - - x x x x

[0079] * QS: Quantum satis

[0080] Referring to Table 1 above, it was confirmed that Compositions 1 and 2, which applied acetaminophen and bromhexine along with florfenicol, were stable at 6 and 8 months under harsh conditions, respectively. This suggests that the content remains stable for approximately 24 months under room temperature conditions (25℃, relative humidity 60%). On the other hand, Composition 3, which applied aspirin and vitamin C along with florfenicol, applied methylpyrrolidone in the same way as Composition 2, but vitamin C decreased by 50%, and aspirin was also evaluated as unstable. In addition, Composition 4, which applied florfenicol and aspirin together, also showed a rapid decrease in aspirin content, indicating poor stability of the composition.

[0081] For Composition 5, vitamin C remained stable for 2 weeks under accelerated conditions for florfenicol, aspirin, and vitamin C, but florfenicol decreased at a faster rate than other compositions, and aspirin content decreased rapidly, making it unstable. For Composition 6, it was confirmed that florfenicol decreased rapidly and aspirin content decreased rapidly after 2 weeks under harsh conditions.

[0083] Reflecting the above experimental results, Composition 1 and Composition 2 were evaluated as excellent for implementation in terms of manufacturing and stability as example compositions, and in the following experiments, Composition 1 was applied as the example composition.

[0085] 2. Efficacy and safety test against chicken E. coli infection

[0086] As an example composition, a composition comprising 200 parts by weight of florfenicol, 200 parts by weight of acetaminophen, and 10 parts by weight of bromhexine hydrochloride as active ingredients based on 1,000 parts by weight of a liquid composition was applied.

[0088] 1) Clinical efficacy

[0089] After purchasing hatching eggs produced from broiler breeder chickens and hatching chicks, from biological samples such as feces (tears, nasal exudate, etc.) E. coli 94 healthy animals in which no [unclear] was detected (20 in the negative control group, 37 in the positive control group, and 37 in the 1x dose group) were selected as the target animals.

[0090] Test group Processing details Clinical indicators and observations Negative control group No infection and no treatment Weight gain rate, feed intake, hematological values, etc. positive control group E. coli Infection + Non-drug group Adjust the bacterial suspension to 1×10^7 CFU / mL and infuse 0.3 mL into the trachea. Confirmation of bacterial isolation, clinical symptoms (weight gain, feed intake, poor gait, diarrhea, etc.), hematological values, gross pathological findings 1x dose administration group E. coli Infection + Drug Treatment Group After confirming respiratory / gastrointestinal symptoms, administer the drug orally at a dose of 0.1 mL per kg of body weight for 5 days (drinking water). Confirmation of bacterial isolation, clinical symptoms (weight gain, feed intake, poor gait, diarrhea, etc.), hematological values, gross pathological findings

[0091] - Administration of test drug

[0092] For 7 days prior to the start of the test, the water intake of the test animals was measured to calculate the average daily water intake per individual, and a test substance at a concentration of 0.26 mL / L was prepared and administered via drinking water for 5 days so that 0.1 mL of the test substance could be consumed per kg of body weight.

[0093] - Weight gain, feed intake, and feed efficiency

[0094] Body weight was measured once before drug administration, daily for 3 days after drug administration, and thereafter at 3-day intervals. Changes in feed intake / conversion ratio were evaluated by measuring the residual amount of feed intake once a day during the test period and using the trough method.

[0095] During the test period, the average daily feed intake and weight gain of the test substance administration group increased statistically significantly compared to the positive control group (p<0.05), but showed no significant difference compared to the negative control group. The feed efficiency of the test substance administration group was higher than that of both the positive and negative control groups, confirming that feed efficiency had improved.

[0096] Test group Before administration 14 days after the start of administration Weight gain (g) 11-day-old 25 days old Negative control group 310±25.1 815±32.1 505.2±7.9a positive control group 271±21.9 722±36.1 437.6±21.0b 1x dose administration group 304±28.9 783±32.6 484.8±38.1a

[0097] Test group Average daily feed intake per head (g) feed efficiency Negative control group 80.1±15.3 6.31 positive control group 73.0±14.4 6.00 1x dose administration group 75.4±14.3 6.43

[0098] - Observation of clinical symptoms

[0099] Clinical symptoms were observed daily and scored (0=normal, 1=poor walking, 2=slight depression or ataxia, 3=severe depression and ataxia).

[0100] The clinical symptom observation index of the 1x dose group decreased statistically significantly from the 3rd day after the start of test drug administration to the 14th day after the start of test drug administration compared to the positive control group.

[0101] (days) after the start of test drug administration Test group 1 (12 days old) 2 (13 days old) 3 (14 days old) 4 (15 days old) 5 (16 days old) Negative control group 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00 positive control group 2.49±0.66 2.43±0.68 2.28±0.68 2.20±0.70 2.20±0.70 1x dose administration group 2.34±0.64 2.20±0.71 1.84±0.47 Q 1.60±0.68 Q 1.25±0.64R (days) after the start of test drug administration Test group 6 (17 days old) 7 (18 days old) 8 (19 days old) 9 (20 days old) 10 (21 days old) Negative control group 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00 positive control group 2.15±0.67 2.13±0.52 2.07±0.46 2.07±0.46 1.90±0.32 1x dose administration group 0.95±0.51R 0.53±0.52R 0.53±0.52R 0.40±0.51R 0.40±0.52R (days) after the start of test drug administration Test group 11 (22 days old) 12 (23 days old) 13 (24 days old) 14 (25 days old) - Negative control group 0.00±0.00 0.00±0.00 0.00±0.00 0.00±0.00 - positive control group 1.80±0.42 1.60±0.52 1.20±0.45 1.20±0.45 - 1x dose administration group 0.30±0.48R 0.10±0.32R 0.00±0.00Q 0.00±0.00Q -

[0102] Clinical observation index: 0=Normal, 1=Does not walk well, 2=Slight depression or ataxia, 3=Severe depression and ataxia.

[0103] Qp<0.01, Rp<0.001, compared with the positive group.

[0105] - Hematological and blood biochemical analysis

[0106] At the end of the experiment, blood was collected from all experimental systems and analyzed for hematological indicators (WBC, RBC, hemoglobulin, MCV, MCHC, platelet, etc.) and blood biochemical indicators (ALT, AST, ALP, BUN, etc.) using an automated blood analyzer.

[0107] In the case of hematological indicators, compared to the positive control group, WBC (p<0.01), MCH (p<0.01), and MCHC (p<0.05) in the 1x dose group decreased statistically significantly, while hemoglobin and platelets increased statistically significantly (p<0.05).

[0108] In the case of blood biochemical markers, compared to the positive control group, ALT (p<0.01), AST (p<0.01), uric acid (p<0.001), and creatinine (p<0.001) in the 1x dose group were statistically significantly decreased, while ALP was statistically significantly increased (p<0.001).

[0109] Hematological parameters Test group WBC(×10^3 / μL) RBC(×10^6 / μL) Hb(g / dL) Hemat(%) MCV(fL) Negative control group 21.16±1.22 3.14±0.26 10.53±0.78 32.83±1.07 102.5±5.4 positive control group 26.92±1.07 2.65±0.29 8.96±0.55 30.76±1.54 103.2±4.4 1x dose group 20.94±1.14** 2.97±0.34 9.85±0.56* 32.16±1.24 102.4±5.6 Hematological parameters Test group MCH(pg) MCHC(g / dL) PLT(×10^9 / μL) - - Negative control group 28.37±1.22 28.78±1.34 12.21±1.06 - - positive control group 33.43±1.34 32.53±1.35 10.14±0.69 - - 1x dose group 28.63±1.55** 29.00±1.40* 12.08±0.98* - - Blood biochemical parameters (Serum biochemical parameters) Test group ALT (U / L) AST (U / L) ALP (U / L) UA (mg / dL) CREA (mg / dL) Negative control group 63.55±3.48 7.42±1.39 311.7±13.4 5.25±0.35 1.16±0.06 positive control group 78.06±3.26 13.20±1.56 263.8±12.0 7.58±0.51 1.96±0.12 1x dose group 65.18±2.14** 8.27±1.25** 305.1±11.5*** 5.42±0.39*** 1.22±0.23*** WBC, white blood cells; RBC, red blood cells; Hb, hemoglobin; Hemat, hematocrit; MCV, mean copuscular volume; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; PLT, platelets; ALT, alanine aminotransferase; AST, aspartate aminotransferase;ALP, alkaline phophatase; UA, uric acid; CREA, creatinine.*p<0.05, **p<0.01, ***p<0.001, compared with the positive group.

[0110] - Observation indices and pathological findings by organ

[0111] Gross and clinical-pathological findings were observed and recorded for animals that died during the experiment and for test animals at the time of completion.

[0112] Air sacs: 0-Normal, 1-Air sac opacity, 2-Air sac wall thickening, 3-One air sac with cheesy exudate and muscular wall, 4-Two or more air sacs with cheesy exudate and muscular wall

[0113] Heart: 0-Normal, 1-Presence of excessively clear or turbid fluid in the pericardial cavity, 2-Extensive fibrinous effusion of the pericardium

[0114] Liver: 0-Normal, 1-Focal fibrin exudate on the surface of the liver, 2-Extensive fibrin exudate

[0115] On the 6th day after the start of the test drug administration, the gross indices for the air sacs, heart, and liver in the 1x dose treatment group were all statistically significantly reduced compared to the positive control group (p<0.05). Subsequently, the gross indices for the air sacs, heart, and liver in the 1x dose treatment group were reduced compared to the positive control group, but no statistically significant difference was observed.

[0116] - Long-term scores Date of autopsy after start of medication Test group envelope heart liver 1 day (12 days old) positive control group 1.00±0.00 0.80±0.45 0.80±0.45 1x dose group 0.60±0.55 0.60±0.55 0.40±0.55 2 (13 days old) positive control group 1.20±0.45 1.00±0.00 0.80±0.45 1x dose group 0.60±0.55 0.62±0.55 0.40±0.55 3 (14 days old) positive control group 1.40±0.55 1.00±0.00 0.80±0.45 1x dose group 0.40±0.55 0.40±0.55 0.40±0.55 6 (17 days old) positive control group 1.20±0.45 1.00±0.00 0.80±0.45 1x dose group 0.20±0.45* 0.20±0.45* 0.20±0.45* 9 days (20 days old) positive control group 0.80±0.45 0.60±0.55 0.40±0.55 1x dose group 0.00±0.00* 0.00±0.00 0.00±0.00 12 days (23 days old) positive control group 0.40±0.55 0.20±0.45 0.00±0.00 1x dose group 0.00±0.00 0.00±0.00 0.00±0.00 14 days (25 days old) positive control group 0.20±0.45 0.00±0.00 0.00±0.00 1x dose group 0.00±0.00 0.00±0.00 0.00±0.00 *p<0.05, compared with the positive control group.

[0117] - Confirmation of bacterial isolation

[0118] After the autopsy for each period, the liver, heart, and spleen are aseptically collected. E. coli As a result of confirming the bacterial count, from day 3 to day 12 after the start of test drug administration, in the heart, liver, and spleen of the 1x dose treatment group E. coli The number of bacteria decreased statistically significantly compared to the positive control group.

[0119] E. coli Bacterial count (CFU / g) (days) after the start of medication positive control group 1x dose group heart liver spleen heart liver spleen 1 32.20±4.76 60.80±7.40 49.00±5.87 25.40±3.97 46.80±5.31 39.60±4.51 2 51.60±5.27 87.20±7.05 67.80±6.69 40.20±4.76 75.20±6.98 54.20±6.06 3 72.20±7.09 122.6±12.8 83.20±7.29 49.40±4.56* 88.60±6.23* 69.80±6.69* 6 53.60±6.88 86.20±8.14 54.80±5.50 35.20±5.63** 57.60±7.33* 43.60±4.88* 9 25.00±3.54 47.40±4.51 37.60±3.97 14.20±3.27* 29.60±3.58** 23.80±4.76** 12 14.00±1.58 25.40±2.88 17.00±2.74 5.20±1.64** 9.80±2.39** 7.60±1.82** 14 4.80±4.09 6.80±5.26 5.40±4.62 0.00±0.00 0.20±0.45 0.00±0.00 *p<0.05, **p<0.01, compared with the positive control group.

[0120] - Confirm removal of respiratory secretions

[0121] After autopsy at each stage, the phenol red effusion method was performed to measure absorbance to confirm the effect of removing secretions from the respiratory tract. As a result, from the 3rd day to the 12th day after the start of drug administration, the absorbance of the drug-treated group decreased statistically significantly compared to the positive control group, confirming that the drug treatment is effective in removing secretions from the respiratory tract.

[0122] Absorbance (mean±SD) after the start of test drug administration (day), *P<0.05, **p<0.01 (days) after the start of medication administration 1 2 3 - positive control group 0.223±0.043 0.292±0.041 0.385±0.058 - 1x dose group 0.213±0.063 0.250±0.049 0.281±0.052 * - Absorbance (mean±SD) after the start of test drug administration (day), *P<0.05 (days) after the start of medication administration 6 9 12 14 positive control group 0.560±0.089 0.447±0.069 0.361±0.056 0.276±0.041 1x dose group 0.246±0.047 ** 0.229±0.049 ** 0.221±0.044 * 0.218±0.031

[0123] - Observe rectal temperature

[0124] As a result of measuring rectal temperature before autopsy at each time point, from the 3rd day to the 12th day after the start of drug administration, the rectal temperature of the drug-treated group decreased statistically significantly compared to the positive control group (p<0.05), confirming that the drug treatment had a body temperature-lowering effect.

[0125] After the start of test drug administration (days), °C (mean±SD), *P<0.05 (days) after the start of medication administration 1 2 3 - positive control group 41.70±0.16 41.98±0.22 42.22±0.24 - 1x dose group 41.64±0.18 41.82±0.08 41.78±0.19 * - After the start of test drug administration (days), °C (mean±SD), *P<0.05 (days) after the start of medication administration 6 9 12 14 positive control group 42.28±0.26 42.18±0.24 41.74±0.27 41.36±0.21 1x dose group 41.62±0.23 * 41.26±0.26 * 41.22±0.13 * 41.18±0.22

[0126] 2) Safety test

[0127] Test group Processing details Clinical indicators and observations Negative control group No infection and no treatment Weight gain rate, feed intake, hematological values, etc. 3x dose administration + 3x drug administration Administer the drug orally at a dose of 0.2 mL per kg of body weight for 5 days (drinking water) Weight gain rate, feed intake, hematological values, etc.

[0128] - Administration of test drug

[0129] For 7 days prior to the start of the test, the water intake of the test animals was measured to calculate the average daily water intake per individual, and a test substance at a concentration of 0.78 mL / L was prepared and administered via drinking water for 5 days so that 0.3 mL of the test substance could be consumed per kg of body weight.

[0131] - Weight changes and feed intake

[0132] Changes in body weight of broiler chickens after the start of administration of the test drug are shown in Table 12, and feed intake and feed efficiency (P<0.05) of broiler chickens after the start of administration of the test drug are shown in Table 13.

[0133] Test group 11 days prior to administration 14 days after the start of administration, 25 days old Weight gain (g) Negative control group 310±25.1 815±32.1 505.2±7.9 3x dose group 313±24.2 828±31.6 514.8±11.0

[0134] Test group Average daily feed intake per head (g) feed conversion ratio Negative control group 80.1±15.3 0.16 1x dose administration group 81.2±15.8 0.16

[0135] - Hematological and blood biochemical analysis results

[0136] The results of hematological and blood biochemical analyses after the initiation of triple dose administration are shown. For both hematological and blood biochemical indicators, none of the indicator values ​​in the triple dose administration group showed statistically significant differences compared to the negative control group.

[0137] Hematological parameters Test group WBC(×10^3 / μL) RBC(×10^6 / μL) Hb(g / dL) Hemat(%) MCV(fL) Negative control group 21.16±1.22 3.14±0.26 10.53±0.78 32.83±1.07 102.5±5.4 3x dose group 21.12±1.28 3.17±0.20 10.68±0.67 32.79±1.13 101.6±3.7 Hematological parameters Test group MCH(pg) MCHC(g / dL) PLT(×10^9 / μL) - - Negative control group 28.37±1.22 28.78±1.34 12.21±1.06 - - 3x dose group 28.52±1.06 28.92±1.26 12.28±1.13 - - Blood biochemical parameters (Serum biochemical parameters) Test group ALT (U / L) AST (U / L) ALP (U / L) UA (mg / dL) CREA (mg / dL) Negative control group 63.55±3.48 7.42±1.39 311.7±13.4 5.25±0.35 1.16±0.06 3x dose group 63.79±3.31 7.62±1.29 313.7±14.3 5.21±0.37 1.14±0.07 WBC, white blood cells; RBC, red blood cells; Hb, hemoglobin; Hemat, hematocrit; MCV, mean copuscular volume; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; PLT, platelets; ALT, alanine aminotransferase; AST, aspartate aminotransferase;ALP, alkaline phophatase; UA, uric acid; CREA, creatinine.*p<0.05, **p<0.01, ***p<0.001, compared with the positive group.

[0138] - Gross and pathological findings

[0139] Grossly, no particular abnormalities were found in the 3x dose group compared to the negative control group.

[0140] Therefore, the composition of the example was confirmed to be a highly effective and safe substance for treating E. coli infection in broiler chickens infected with E. coli, the causative agent of broiler E. coli infection.

[0142] Although preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements by those skilled in the art using the basic concept of the present invention as defined in the following claims also fall within the scope of the present invention.

Claims

Claim 1 A veterinary composition for alleviating or inhibiting E. coli in poultry, comprising florfenicol, acetaminophen, and bromhexine as active ingredients, wherein the composition comprises 10 to 30 parts by weight of florfenicol and 10 to 30 parts by weight of acetaminophen based on 1 part by weight of bromhexine, and further comprises any one additive selected from the group consisting of dimethylacetamide, methylpyrrolidone, dimethylformamide, and combinations thereof; and any one excipient selected from the group consisting of propylene glycol, polyethylene glycol, glycerin, and combinations thereof. Claim 2 delete Claim 3 delete Claim 4 delete Claim 5 delete Claim 6 A veterinary composition for alleviating or inhibiting E. coli in poultry, wherein, in claim 1, the number of E. coli in poultry is reduced three days after the start of administration. Claim 7 A veterinary composition for alleviating or inhibiting E. coli in poultry according to claim 1, comprising 0.7 to 1.3 parts by weight of acetaminophen based on 1 part by weight of florfenicol. Claim 8 A method of administering a veterinary composition for alleviating or inhibiting E. coli in poultry according to claim 1, by mixing the veterinary composition for alleviating or inhibiting E. coli in poultry with drinking water and orally administering the bromhexine at a concentration of 0.5 to 2 mg per 1 kg of body weight of the poultry individual. Claim 9 A method of administering a veterinary composition for alleviating or suppressing E. coli in poultry, wherein, in comparison to poultry infected with E. coli, poultry infected with E. coli after the start of administration shows increased weight gain and improved feed requirements. Claim 10 In claim 8, a method of administering a veterinary composition for alleviating or inhibiting E. coli in poultry, having antipyretic and expectorant effects on poultry with E. coli.