High-activity anti-coccidiosis nicarbazin composition and preparation method thereof

Abstract

The invention discloses a high-activity anti-coccidiosis nicarbazin composition, which relates to the technical field of poultry medicines and comprises the following components in percentage by mass: 8 to 40 percent of nicarbazin, 0.1 to 8 percent of polycarbophil or polycarbophil salt, 0.5 to 10 percent of antibacterial peptide, 0.2 to 6 percent of PQQ and the balance of diluent. The composition can prolong the residence time of the medicine in the intestinal tract, meanwhile, the composition is beneficial to decomposition of nicarbazin into more 4, 4 '-dinitrosym-diphenylurea (DNC) crystals with superfine particle sizes in the intestinal tract, the nicarbazin can more easily act on coccidiosis parasitizing in the intestinal tract or be absorbed into blood, the problem of the coccidiosis is efficiently solved, and meanwhile the effect of the nicarbazin on the intestinal tract is achieved. The traditional Chinese medicine composition can assist in controlling secondary clostridial enteritis, and can more effectively maintain intestinal health of poultry by controlling two pathogenic factors, namely coccidiosis and clostridium, which are the most important pathogenic factors for destroying intestinal tracts in chicken raising.

Description

Technical Field

[0001] The invention relates to the technical field of poultry drugs, and in particular to a high-activity anticoccidial nicarbazin composition and a preparation method thereof. Background Art

[0002] In the poultry industry, coccidiosis is a common and high-impact disease. This pathogen can cause stunted growth, reduced egg production, and even death of chickens, resulting in huge economic losses to breeders.

[0003] At present, the main means of preventing and controlling chicken coccidiosis are the use of anticoccidial drugs and vaccination. Although there is no problem with coccidiosis vaccine in controlling clinical coccidiosis, it will inevitably cause certain intestinal damage during the oocyst circulation infection period required for immunity, and it is more likely to develop clostridial enteritis, which will affect the health of the intestine. Nicarbazin exerts a better anticoccidial effect by producing ultrafine 4,4'-dinitrocarbamide (DNC) crystals in the gastrointestinal tract. However, different formulation process factors will affect the amount of ultrafine DNC crystals produced and the residence time of DNC crystals in the intestine, which in turn affects the efficacy.

[0004] To this end, the present invention is specially proposed to solve the above-mentioned technical problems. Summary of the invention

[0005] The object of the present invention is to provide a highly active anticoccidial nicarbazin composition, comprising the following components in mass percentage: nicarbazin 8%-40%, polycarbophil or its salt 0.1%-8%, antimicrobial peptide 0.5%-10%, pyrroloquinoline quinone (PQQ) 0.2%-6%, and the balance being a diluent.

[0006] Preferably, the mass percentage of nicarbazine is 15% to 30%.

[0007] Preferably, the mass percentage of the polycarbophil or its salt is 0.5% to 5%.

[0008] Preferably, the antimicrobial peptide is selected from one or more of heptacopeptide subtilisin, cecropin, gramicidin and bacitracin.

[0009] Preferably, the antimicrobial peptide is selected from heptathriatin subtilisin.

[0010] Preferably, the mass percentage of the antimicrobial peptide is 1% to 5%.

[0011] Preferably, the mass percentage of the pyrroloquinoline quinone (PQQ) is 0.5% to 3%.

[0012] Preferably, the diluent is one or more of corn cob powder, defatted rice bran, residues of the extract of Artemisia annua and Changshan granules, or residues of the extract of Artemisia annua and Changshan granules.

[0013] Preferably, the composition is a premix.

[0014] A method for preparing a highly active anticoccidial nicarbazin composition, characterized in that it comprises the following steps: (1) Mix polycarbophil, antimicrobial peptide, pyrroloquinoline quinone (PQQ) and a small amount of diluent; (2) Add nicarbazine and mix well; (3) Add the remaining diluent, mix well and package.

[0015] The beneficial effects of the present invention are: 1. The antimicrobial peptides in the composition of the present invention have a broad-spectrum antibacterial effect, have a certain effect on Clostridium perfringens, can inhibit and kill the growth of some harmful bacteria, and promote intestinal health. For example, subtilisin 37 peptide is super stable and has good resistance to high temperature, acid, alkali, pepsin, and trypsin. Subtilisin 37 peptide can improve the immune performance of broilers by promoting lymphocyte proliferation, improving immune organ index, serum antibody level, etc.; subtilisin 37 peptide can increase beneficial bacteria such as lactobacillus and bifidobacterium, kill harmful bacteria such as intestinal Escherichia coli, Salmonella, Clostridium perfringens, regulate intestinal flora, maintain the stability of intestinal microecological environment, reduce animal diarrhea, and improve intestinal barrier function by regulating the balance of intestinal flora, thereby improving intestinal health, feed conversion rate and growth performance of broilers.

[0016] 2. The (pyrroloquinoline quinone) PQQ in the composition of the present invention has multiple physiological functions such as anti-oxidation, anti-inflammation, regulation of immunity and energy metabolism, and regulation of cell signaling pathways, and can cooperate with antimicrobial peptides to improve the growth performance and health level of broilers.

[0017] 3. The polycarbophil in the composition of the present invention can absorb water, which is retained in the intestine, helping nicarbazine to produce more 4,4'-dinitrocarbamide (DNC) in the intestine. Whether it acts on coccidia through absorption into the blood or directly acts on coccidia parasitic on the intestine, it can more efficiently exert an anti-coccidial effect; at the same time, the polycarbophil in the composition has bioadhesive properties, can interact with mucosal proteins, increase the retention time of the preparation in the intestine, and increase its absorption. The composition of the present invention has no effect on the water metabolism of the intestine and can also reduce the frequency of diarrhea.

[0018] 4. The composition of the present invention can effectively solve the problem of coccidia and assist in controlling the secondary clostridial enteritis problem. By controlling coccidia and clostridium, the two most important pathogenic factors that damage the intestines in chicken farming, the intestinal health of poultry can be more effectively maintained. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 It is the blood concentration curve of DNC and HDP after single dose administration; Figure 2 It is a bar graph of drug content in intestinal tissue after single-dose administration. DETAILED DESCRIPTION

[0020] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0021] Example 1 A method for preparing a highly active anticoccidial nicarbazin composition is as follows: weigh 5g of polycarbophil, 5g of heptapeptide subtilisin, 1g of pyrroloquinoline quinone (PQQ), and 20g of corn cob powder, and mix them for 5 minutes; add 25g of nicarbazin, and mix them for 7 minutes; add 44g of corn cob powder, and mix them for 10 minutes to obtain the composition.

[0022] Example 2 A preparation method of a highly active anticoccidial nicarbazine composition is as follows: weigh 5g of polycarbophil, 5g of heptapeptide-37 subtilisin, 1g of pyrroloquinoline quinone (PQQ), and 20g of the residue of an extract of Artemisia annua and Changshan granules (after drying and crushing), and mix for 5min; add 25g of nicarbazine, and mix for 7min; add 44g of the residue of an extract of Artemisia annua and Changshan granules (after drying and crushing), and mix for 10min.

[0023] Comparative Example A method for preparing an anticoccidial nicarbazin composition is as follows: weigh 25 g nicarbazin and 25 g corn cob powder, and mix for 5 minutes; add 50 g corn cob powder, and mix for 15 minutes.

[0024] The anticoccidial effects of Examples 1 and 2 and the comparative example are tested below. 260 1-day-old broiler chickens were raised in a strictly disinfected chicken house, with free access to food and water, and raised to 10 days of age. The feces microscopic examination of chicken coccidia was negative. The test chickens were weighed one by one on an empty stomach in the morning of 10 days of age, and the weak and oversized ones were eliminated. Then, they were randomly divided into 5 groups according to the same initial weight, with 25 in each group, namely: a blank control group (no infection and no treatment), a control group (infection and no treatment), a comparative group, an embodiment 1 group, and an embodiment 2 group. Anti-coccidial drugs were fed for 3 days from the 11th day of age, and no drugs were fed on the 14th day of age. On the 15th day, except for the blank control group, 50,000-60,000 sporulated oocysts of Eimeria tenella were inoculated in the other groups. 3 days after the attack, the comparative group and the nicarbazin composition group of the present invention were fed with nicarbazin 125ppm mixed in the feed. After inoculation of coccidia oocysts, observe the mental state, feeding, mortality, and fecal color of each group of chickens every day and keep records; weigh each chicken on an empty stomach on the first and tenth day after infection, and calculate the weight gain of each group, and then calculate the average weight gain and relative weight gain rate. On the tenth day after infection, 9 chickens in each group were killed, and the cecal oocysts were counted and converted into oocyst values, and the lesion scores were scored and converted into lesion values. The anticoccidial index (ACI) recommended by Merck Company of the United States was used as an indicator to determine the efficacy of the drug: ACI = (survival rate + relative weight gain rate) × 100 - (lesion value + oocyst value) ACI>180 is judged as a highly effective anticoccidial drug; ACI of 160~180 is judged as a medium-effective anticoccidial drug; ACI of 120~160 is judged as a low-efficiency anticoccidial drug; ACI<120 is judged as an ineffective anticoccidial drug. The results are shown in Table 1 Table 1 Anticoccidial effect of different groups As shown above, the anticoccidial effects of Examples 1 and 2 are higher than those of the comparative example group, and they are highly effective anticoccidial drugs.

[0025] Next, the effect test of necrotic enteritis was carried out on Example 2. 120 9-day-old white-feathered broilers with similar body weight and good health were randomly divided into 2 groups: Group 1: comparative example; Group 2: Example 2, with 3 replicates in each group and 20 broilers in each replicate for testing. 9-20 days old: Group 1 (comparative example) was fed a diet containing the comparative example (measured at a concentration of 125 ppm of nicarbazin); Group 2 (Example 2) was fed a composition containing this example (added to the previous diet at a concentration of 125 ppm of nicarbazin). 17-20 days old, each group was gavage-fed daily with Clostridium perfringens (containing 2.0*10 8CFU / ml live bacteria, 1ml / animal). From the 21st day of age, the front-end diet of all groups was changed to the back-end diet. After the challenge was completed at the 20th day of age, 8 animals were randomly selected from each replication, that is, 24 animals were selected from each group. After slaughter and dissection, the complete small intestine sample was taken and scored according to the severity of necrotic enteritis. The results are as follows: Table 2 Scoring table for necrotic enteritis Note: 2 points or above are considered as onset.

[0026] As can be seen from the above table, the therapeutic effect of Example 2 on necrotic enteritis is better than that of the comparative example group.

[0027] Using Example 2 to investigate the plasma concentration of a single dose Sixteen 21-day-old white-feathered broilers with similar body weight and good health were selected and randomly divided into two groups, with 8 in each group. The first group was Example 2; the second group was the comparative group. The chickens were fed with a single dose of 125 mg (nicarbazine) / kg feed after oral administration, and blood was collected at 1, 2, 4, and 24 hours, respectively, and the blood samples were collected for analysis.

[0028] Table 3 Plasma concentrations of DNC and HDP after single-dose administration Note: In the table, DNC is 4,4'-dinitrophenylurea; HDP is 2-hydroxy-4,6-dimethylpyrimidine.

[0029] As can be seen from Table 3, the blood drug concentrations of Example 2 of the present invention at 1, 2, 4, and 24 hours were higher than those of the comparative example group, indicating that the in vivo absorption and drug concentration maintenance time of Example 2 were better than those of the comparative example group.

[0030] Using Example 2 to investigate the concentration of HDP and DNC in intestinal tissue Sixteen 21-day-old white-feathered broiler chickens with similar body weight and good health were selected and randomly divided into 2 groups (Example 2 group and comparative example group), 8 in each group, and the chickens were fed with a single dose of 125 mg (nicarbazine) / kg feed after oral administration. The chickens were slaughtered 4 hours after administration, the small intestine was taken, chopped and homogenized, the homogenized test sample was taken, solvent was added for dissolution and extraction, centrifuged, and analyzed by high performance liquid chromatography.

[0031] Table 4 Drug content in the intestine after single dose administration It can be seen from Table 4 that, under the same conditions, the drug content of the sample of Example 2 in the intestinal tissue is higher than that of the comparative example group, indicating that the sample of Example 2 has better absorption in the body.

[0032] From the test results in Tables 1 to 4, it can be seen that the compositions of Examples 1 and 2 of the present invention are superior to the comparative example products. This is because the compositions of the present invention have bioadhesive properties, which can prolong the retention time of the drug in the intestine, promote absorption, and have a good therapeutic effect on necrotic enteritis caused by Clostridium.

[0033] In addition, a broiler farm was mixed-fed with the product of Example 2 (calculated as nicarbazine, 125 g / 1000 kg feed) and slaughtered 4 days after drug withdrawal. The residual amount of the residual marker 4,4'-dinitrourea in the edible tissues was measured. The results showed that the residual limits of the residual markers in each target tissue were all lower than the national residual limit standard of 200 μg / kg, and would not cause the residual drugs to exceed the standard.

[0034] Although the embodiments of the present application have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present application, and that the scope of the present application is defined by the appended claims and their equivalents.

Claims

1. A highly active anticoccidial nicarbazin composition, characterized in that: The composition comprises the following components in mass percentage: 8%-40% of nicarbazine, 0.1%-8% of polycarbophil or its salt, 0.5%-10% of antimicrobial peptide, 0.2%-6% of PQQ, and the balance is diluent.

2. A highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The mass percentage of the nicarbazine is 15% to 30%.

3. A highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The mass percentage of the polycarbophil or its salt is 0.5% to 5%.

4. A highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The antimicrobial peptide is selected from one or more of heptacopeptide subtilisin, cecropin, gramicidin and bacitracin.

5. A highly active anticoccidial nicarbazin composition according to claim 4, characterized in that: The antimicrobial peptide is selected from Heptacopeptide subtilisin.

6. A highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The mass percentage of the antimicrobial peptide is 1% to 5%.

7. A highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The mass percentage of the PPQ is 0.5% to 3%.

8. A highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The diluent is one or more of corn cob powder, defatted rice bran, residues of extract of Artemisia annua and Changshan granules or residues of extract of Sanwei Kangqiu granules.

9. A highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The composition is a premix.

10. The method for preparing a highly active anticoccidial nicarbazin composition according to claim 1, characterized in that: The following steps are involved: (1) Mix polycarbophil, antimicrobial peptide, PQQ and a small amount of diluent evenly; (2) Add nicarbazine and mix well; (3) Add the remaining diluent, mix well and package.

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