Pharmaceutical composition for preventing and treating calf diarrhea

Abstract

The invention discloses a pharmaceutical composition for preventing and treating calf diarrhea. The pharmaceutical composition is prepared from 0.8 g of enterobacter peptide and 0.05-0.2 g of purslane extract. According to the clinical manifestation of the calf, a proper combination ratio is selected and added into the feeding milk of the calf, so that the excrement score and diarrhea rate of the calf can be reduced. The composition improves the immunity of calves by increasing immune globulins IgA, IgG and IgM, and regulates the health state of intestinal tracts by reducing the number of escherichia coli in the intestinal tracts. By reducing inflammatory factors IL-1beta and IL-6, inflammatory response in calves is reduced, the cure rate of diseased cattle is increased, and meanwhile the average cure day number is reduced.

Description

Technical Field

[0001] The invention belongs to the field of veterinary medicines, and particularly relates to a pharmaceutical composition for preventing and treating calf diarrhea. Background Art

[0002] Calf diarrhea is one of the common diseases in the cattle industry. It is often caused by pathogenic microorganism infection, untimely colostrum feeding, poor feeding environment, sudden weather changes or sudden changes in diets. After the onset of the disease, it is easy to cause dehydration, acidosis, systemic nutritional disorders and other symptoms. The incidence rate is about 80%. It is a common disease that endangers the health of calves and causes serious economic losses to farms. It is more common in calves under 30 days old. The disease not only affects the normal growth and development of calves, but also seriously affects the reproductive capacity and lactation performance of dairy cows, and then causes huge losses to the cattle industry.

[0003] In veterinary clinics, the main methods for preventing and treating calf diarrhea are vaccination, strengthening feeding management, and using antibiotics. However, the effect of vaccination is unstable, the cost of feeding management is high, and excessive use of antibiotics can easily lead to more serious bacterial resistance problems, reduced clinical treatment effects, and increased risk of veterinary drug residues in livestock products, posing a major threat to animal health and agricultural production. Therefore, it is of great practical significance to develop a safe and effective drug combination for preventing and treating calf diarrhea.

[0004] Traditional Chinese medicine is widely used in the medical and animal husbandry industries. It is favored by researchers for its wide range of antibacterial activity and low residual toxicity. Studies have shown that traditional Chinese medicine can promote the growth of beneficial bacteria in the rumen of calves, inhibit the growth of harmful bacteria, promote the digestion and utilization of nutrients in feed by calves, improve the body's metabolic level, enhance immunity, and improve disease resistance.

[0005] Microcin J25 is a natural bacterial antimicrobial peptide with high safety and stable properties. It can regulate intestinal flora and improve the body's immunity. It has a killing effect on pathogens of calf diarrhea, such as Escherichia coli and Salmonella. Compared with traditional antibiotics, pathogens are less likely to develop resistance to microcin. However, experiments have shown that microcin J25 alone has certain limitations when used to prevent and treat calf diarrhea and cannot achieve the expected effect. Summary of the invention

[0006] In view of the problems existing in the above-mentioned prior art, the object of the present invention is to provide a pharmaceutical composition for preventing and treating calf diarrhea. The combination combines enterobactin with purslane extract, which can effectively prevent the incidence of calf diarrhea and improve the cure rate.

[0007] To achieve the above object, the technical solution adopted by the present invention is: a pharmaceutical composition for preventing and treating calf diarrhea, which is composed of 0.8g of enterobactin and 0.05g-0.2g of purslane extract.

[0008] Furthermore, the optimal addition amount of the pharmaceutical composition for preventing and treating calf diarrhea of ​​the present invention is 0.8g of enterobactin and 0.1g of purslane extract.

[0009] Furthermore, the purslane extract is a 10:1 concentrated extract.

[0010] Furthermore, the calf is a dairy calf, which is 7 to 15 days old.

[0011] The purslane selected by the present invention is sour and cold; it enters the large intestine and liver meridians; it has the effects of cooling blood and detoxifying, removing dampness and relieving stranguria, and stopping dysentery; it is mainly used to treat diarrhea and dysentery caused by damp heat.

[0012] Another object of the present invention is to provide a pharmaceutical composition for preventing and treating calf diarrhea for use in improving the immunity of healthy calves, regulating intestinal flora and reducing the inflammatory response function in diarrheal calves.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: the pharmaceutical composition provided by the present invention is obtained through screening based on the theoretical basis of traditional Chinese medicine and modern pharmacology, pathogenesis and immunology. The composition mainly achieves the purpose of preventing and treating diarrhea through the mechanism of antibacterial and anti-inflammatory, cooling blood and stopping dysentery, and regulating gastrointestinal function. It has good efficacy, is green, safe, and has no toxic side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Effects of enterobactin and purslane extract on diarrhea rates in calves.

[0015] Figure 2 Effects of enterobactin and purslane extract on serum immunoglobulin IgA in calves.

[0016] Figure 3 Effects of enterobactin and purslane extract on calf serum immunoglobulin IgG.

[0017] Figure 4 Effects of enterobactin and purslane extract on serum immunoglobulin IgM in calves.

[0018] Figure 5 Effects of enterobactin and purslane extract on body temperature in calves.

[0019] Figure 6 Effects of enterobactin and purslane extract on cure rate in calves.

[0020] Figure 7Effects of enterobactin and purslane extract on mean cure days in calves.

[0021] Figure 8 Effects of enterobactin and purslane extract on inflammatory factor IL-1β in calf serum.

[0022] Fig. 9 Effects of enterobactin and purslane extract on inflammatory factor IL-6 in calf serum.

[0023] Fig.10 Effects of enterobactin and purslane extract on inflammatory factor TNF-α in calf serum. DETAILED DESCRIPTION

[0024] The following is a further description of the technical solution of the present invention in combination with the embodiments of the present invention. Obviously, based on the embodiments of the present invention, the embodiments modified in different ways by other persons in the art without creative work should be regarded as the protection scope of the present invention.

[0025] Example 1

[0026] A pharmaceutical composition for preventing calf diarrhea consists of 0.8g of enterobactin and 0.05g of purslane extract.

[0027] Example 2

[0028] A pharmaceutical composition for treating calf diarrhea consists of 0.8g of enterobactin and 0.1g of purslane extract.

[0029] Example 3

[0030] A pharmaceutical composition for treating calf diarrhea consists of 0.8g of enterobactin and 0.15g of purslane extract.

[0031] Example 4

[0032] A pharmaceutical composition for treating calf diarrhea consists of 0.8g of enterobactin and 0.2g of purslane extract.

[0033] The dosage of the pharmaceutical composition for treating calf diarrhea of ​​the present invention is 0.8g / kg of enterobactin and 0.05g-0.2g / kg, wherein g refers to the weight of the drug, kg refers to the weight of the calf, and g / kg represents the dosage per kg of body weight.

[0034] Test Example 1: Determination of combined antibacterial effect

[0035] Preparation of pathogenic bacteria: Three strains of Escherichia coli and three strains of Salmonella with strong pathogenicity were isolated and identified from the feces of clinical calf diarrhea cases. Single colonies were picked and inoculated into TSB broth and cultured at 37°C for 16 to 18 hours before use.

[0036] The checkerboard method was used to evaluate the combined antibacterial effect of enterobactin and purslane extract: the combined antibacterial index FICI of enterobactin and purslane extract against pathogens was determined.

[0037] Table 1 Combined antibacterial effect of enterobactin and purslane extract on Escherichia coli in calf diarrhea

[0038]

[0039] Table 2 Combined antibacterial effect of enterobactin and purslane extract on Salmonella in calf diarrhea

[0040]

[0041] Result analysis:

[0042] Table 1 shows that the FICI of the drug combination of the present invention for the standard strain of Escherichia coli is 0.75, showing a partial synergistic effect, and the other Escherichia coli isolates show partial synergistic and irrelevant effects. Table 2 shows that the FICI of the drug combination of the present invention for the standard strain of Salmonella is 0.25, showing a synergistic effect, and the other Salmonella isolates show partial synergistic and irrelevant effects. This shows that the drug combination has a good synergistic antibacterial effect on calf diarrhea Escherichia coli and Salmonella.

[0043] Test Example 2: Test for determination of calf diarrhea rate

[0044] 80 healthy Holstein calves were selected and randomly divided into 8 groups, with 10 replicates in each group and 1 calf in each replicate. The specific groups are shown in Table 2. The experimental period was 7 days, and each group of calves was fed 4L of pasteurized milk every day. They were given free drinking water, and the calves were subjected to normal disinfection and immunization procedures.

[0045] Table 3 Calf grouping

[0046] Grouping deal with Control group (NC) Feeding normal milk Enterobactin group (MCJ) Normal milk + 0.8g enterobactin Low-dose Purslane group (CHFL) Normal milk + 0.05g purslane extract Medium dose purslane group (CHFM) Normal milk + 0.1g purslane extract High-dose Purslane group (CHFH) Normal milk + 0.2g purslane extract Low-dose combination group (COML) Normal milk + 0.8g enterobactin + 0.05g purslane extract Medium-dose combination group (COMM) Normal milk + 0.8g enterobactin + 0.1g purslane extract High-dose combination group (COMH) Normal milk + 0.8g enterobactin + 0.2g purslane extract

[0047] Note: NC, MCJ, CHFL, CHFM, CHFH, COML, COMM and COMH represent the control group, enterobactin group, low-dose purslane group, medium-dose purslane group, high-dose purslane group, low-dose combination group, medium-dose combination group and high-dose combination group, respectively, which are the same in the following tables and figures.

[0048] During the experiment, the feces characteristics of calves in each group were observed before morning feeding every day, with 0 = firm, 1 = loose and formed feces, 2 = very loose and unformed feces, and 3 = watery feces. Calves with feces scores ≥ 2 for more than two consecutive days were recorded as having diarrhea.

[0049] Figure 1 The effect of enterobactin and purslane extract on calf diarrhea rate

[0050] like Figure 1 As shown in the figure, the diarrhea rate of calves in the control group was the highest, reaching 50%. Compared with the control group, the diarrhea rate of calves supplemented with 0.8g of enterobactin was significantly reduced to 20%. The diarrhea rate of calves supplemented with 0.1g and 0.2g of purslane extract was reduced. When 0.05g, 0.1g, and 0.2g of purslane extract were combined with 0.8g of enterobactin to feed calves, the diarrhea rate was further reduced compared with the control group, especially the combination of 0.1g and 0.2g of purslane extract and 0.8g of enterobactin, with the most significant effect, and the diarrhea rate dropped to 10%.

[0051] As shown in Table 4, adding 0.8g of enterobactin can significantly reduce the fecal score of calves; adding 0.1g and 0.2g of purslane extract can significantly reduce the fecal score of calves; 0.05g, 0.1g, and 0.2g of purslane extract combined with 0.8g of enterobactin can significantly reduce the fecal score of calves.

[0052] Table 4 Effect of combined use of enterobactin and purslane extract on fecal scores of calves

[0053]

[0054] Note: When comparing uppercase and lowercase letters with the same shoulder, an uppercase letter indicates significant difference (P<0.05).

[0055] Test Example 3: Determination of immunoglobulin content and intestinal flora in calves

[0056] The calves were grouped and fed according to the feeding plan of Experimental Example 2. On the morning of the 8th day of the experiment, whole blood was collected from the jugular vein of the calves. The collected blood was coagulated in a centrifuge tube and then centrifuged to obtain serum. The content of immunoglobulins IgA, IgG, and IgM was detected using an ELISA kit.

[0057] Figures 2 to 4 This is a graph showing the effects of enterobactin and purslane extract on calf serum immunoglobulins.

[0058] Note: * indicates significant difference compared with the control group (P < 0.05), ** indicates extremely significant difference (P < 0.01), *** indicates extremely significant difference (P < 0.001); ## indicates extremely significant difference compared with the enterobactin group (P < 0.01).

[0059] like Figure 2 to Figure 4 As shown, 0.05g, 0.1g, and 0.2g of purslane extract combined with 0.8g of enterobactin can significantly increase the content of immunoglobulin IgA in calf serum; 0.1g and 0.2g of purslane extract combined with 0.8g of enterobactin can significantly increase the content of immunoglobulin IgM in calf serum; adding 0.1g and 0.2g of purslane extract, and 0.05g, 0.1g, and 0.2g of purslane extract combined with 0.8g of enterobactin can significantly increase the content of immunoglobulin IgG in calf serum.

[0060] The combination of 0.1 g of purslane extract and 0.8 g of enterobactin can significantly reduce the number of Escherichia coli in calf feces, as shown in Table 5.

[0061] As shown in Table 6, the combination of 0.1 g of purslane extract and 0.8 g of enterobactin can significantly reduce the number of harmful bacteria Escherichia coli in calf feces.

[0062] Table 5 Effect of combined use of enterobactin and purslane extract on the number of Escherichia coli and Lactobacillus in calf feces

[0063]

[0064] Note: When comparing uppercase and lowercase letters with the same shoulder, an uppercase letter indicates significant difference (P<0.05).

[0065] Test Example 4: Determination of calf body temperature, cure rate and average cure days

[0066] Twenty calves with diarrhea were selected and randomly divided into 4 groups, with 5 replicates in each group and 1 calf in each replicate. The specific groups are shown in Table 6. The experimental period was 7 days, and each group of calves was fed 4L of pasteurized milk every day. They were given free drinking water, and the calves were subjected to normal disinfection and immunization procedures.

[0067] Table 6 Calf grouping

[0068] Grouping deal with Enrofloxacin group (ENR) Normal milk + 0.1mL / kg enrofloxacin injection Enterobactin group (MCJ) Normal milk + 0.8g enterobactin Purslane group (CHF) Normal milk + 0.1g purslane extract Combination Group (COM) Normal milk + 0.8g enterobactin + 0.1g purslane extract

[0069] Note: ENR, MCJ, CHF and COM represent enrofloxacin group, enterobactin group, purslane group and combination group, respectively, which are the same in the following tables and figures.

[0070] During the experiment, the body temperature and feces score of calves in each group were observed daily, and calves with feces scores < 2 for more than two consecutive days were recorded as recovered.

[0071] Figure 5 This is a graph showing the effects of enterobactin and purslane extract on body temperature in calves.

[0072] like Figure 5 As shown in the figure, the body temperature of calves supplemented with 0.1 mL / kg enrofloxacin and 0.1 g purslane extract decreased slowly, with some fluctuations. However, the body temperature of calves supplemented with 0.8 g enterobactin and 0.1 g purslane and 0.8 g enterobactin decreased significantly, approaching normal levels on the 6th day.

[0073] Figure 6 This is a graph showing the effects of enterobactin and purslane extract on the cure rate of calf diarrhea.

[0074] like Figure 6 As shown, the cure rate of calves fed with 0.1g of purslane extract combined with 0.8g of enterobactin was significantly higher than that of the other three groups.

[0075] Figure 7 This is a graph showing the effects of enterobactin and purslane extract on the average cure days for calf diarrhea.

[0076] Note: * indicates significant difference (P<0.05), ** indicates extremely significant difference (P<0.01).

[0077] like Figure 7 As shown, the average cure days of calves fed with 0.1g purslane extract combined with 0.8g enterobactin were significantly lower than those of calves fed with 0.1g purslane extract and 0.8g enterobactin alone.

[0078] Test Example 5: Determination of inflammatory factors in calf serum

[0079] The calves were grouped and fed according to the feeding plan of Experimental Example 4. On the morning of the 8th day of the experiment, whole blood was collected from the jugular vein of the calves. The collected blood was coagulated in a centrifuge tube and then centrifuged to obtain serum. The content of inflammatory factors IL-1β, IL-6, and TNF-α was detected using an ELISA kit.

[0080] Figures 8 to 10 This is a graph showing the effects of enterobactin and purslane extract on inflammatory factors in calf serum.

[0081] Note: * indicates significant difference (P<0.05), ** indicates extremely significant difference (P<0.01), *** indicates extremely significant difference (P<0.001).

[0082] like Figures 8 to 10As shown, feeding calves with 0.1g purslane extract and 0.8g enterobactin together can significantly reduce the content of inflammatory factor IL-1β in calf serum; adding 0.1g purslane extract and 0.8g enterobactin alone can significantly reduce the content of serum inflammatory factor IL-6, while feeding calves with 0.1g purslane extract and 0.8g enterobactin together can significantly reduce the content of serum inflammatory factor IL-6.

[0083] From the above examples, we can know that: Bacteriocins and purslane extract have an ideal combined antibacterial effect on calf diarrhea pathogens, such as Escherichia coli and Salmonella; adding 0.1g of purslane extract and 0.8g of Bacteriocins to feed calves can reduce the incidence of calf diarrhea; by increasing the content of immunoglobulins IgA, IgG, and IgM, the immunity of calves is enhanced; by reducing the number of Escherichia coli in the intestine to regulate the health of the intestine. Compared with traditional treatment methods, the combination of Bacteriocins and purslane extract can improve the cure rate of calf diarrhea and significantly reduce the average cure days; by reducing the content of IL-1β and IL-6 in serum, the intestinal inflammatory response is alleviated.

Claims

1. A pharmaceutical composition for preventing and treating calf diarrhea, characterized in that: The combination consists of 0.8 g of enterobactin and 0.05 g to 0.2 g of purslane extract.

2. A pharmaceutical composition for preventing and treating calf diarrhea according to claim 1, characterized in that: The optimal addition amount of the combination is 0.8g of enterobactin and 0.1g of purslane extract.

3. A pharmaceutical composition for preventing and treating calf diarrhea according to claim 1, characterized in that: Purslane extract is a 10:1 concentrated extract.

4. A pharmaceutical composition for preventing and treating calf diarrhea according to claim 1, characterized in that: The calf is a dairy calf, which is 7 to 15 days old.

5. Use of the pharmaceutical composition according to claim 1 in preventing and treating calf diarrhea, improving calf immunity, reducing calf intestinal inflammatory response and regulating intestinal flora.

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