Use of a probiotic formulation for improving udder health in dairy cows

Abstract

The application relates to application of a probiotic preparation in improving dairy cow mammary gland health, and belongs to the technical field of probiotic preparation application. The application finds that a microbial preparation prepared from Akkermansia muciniphila is helpful to maintaining the dairy cow mammary gland health, thereby improving the milk yield and milk quality of the dairy cow, by promoting the proliferation of the dairy cow mammary gland epithelial cells, inhibiting the apoptosis of the dairy cow mammary gland epithelial cells (alleviating mammary gland inflammation and promoting the expression of milk secretion related genes). The preparation is safer, more efficient and pollution-free, is non-toxic and harmless, does not cause negative influence on the health of the dairy cow, and is easy to obtain, easy to culture in a large amount, simple to operate, easy to obtain raw materials and simple in method, so that the cost of manpower and material resources can be reduced.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of application of probiotic preparations, and particularly relates to application of a probiotic preparation in improving udder health of dairy cows. BACKGROUND

[0002] Dairy cow breeding industry is an important part of modern livestock breeding industry, and how to improve the lactation and milk quality of dairy cows has always been the focus of attention in the field of dairy cow breeding. Ensuring the udder health of dairy cows can not only avoid mastitis and reduce the breeding cost, but also ensure the safety and quality of the produced milk. The udder health of dairy cows is closely related to pathogens, environment, feeding and management conditions and many other factors. How to take effective measures to ensure the udder health of dairy cows is of great significance to improve the milk yield and quality of dairy cows.

[0003] As a part of the complex microecological system in the animal body, the intestinal flora not only acts locally but also acts on the whole body, and can affect the metabolism, immune system and inflammatory response of the host, and is closely related to the occurrence and development of various diseases and many diseases of different systems. Current studies have shown that the intestinal flora is closely related to mammary gland diseases and plays a major role in the occurrence, development and treatment of mastitis. Studies have shown that intestinal flora imbalance can lead to the occurrence of mastitis. At the same time, studies have shown that probiotics can treat dairy cow mastitis by regulating intestinal flora. It has been reported that probiotics producing butyrate can reduce the severity of Staphylococcus aureus-induced mastitis, and symbiotic Bacillus can alleviate Staphylococcus aureus-induced mastitis, which suggests that probiotic preparations can be used as a means to promote the udder health of dairy cows. Probiotic preparations refer to active substances that are beneficial to the health of the host when used in appropriate doses, and have the effects of maintaining or adjusting the microecological balance, preventing diseases and improving the health of the host.

[0004] Akkermansia muciniphila (Akk) is a gram-negative oval obligate anaerobic bacterium belonging to the Verrucomicrobia phylum, which was first isolated from healthy human fecal samples by Dutch microbiologist Anton DL Akkermans in 2004 and named after him. Akkermansia muciniphila has been reported to be able to relieve various inflammations such as colitis, hepatitis and periodontitis, but as of now, there is no related report about the application of Akkermansia muciniphila preparation in the udder health of dairy cows. SUMMARY

[0005] Therefore, the application aims to provide an application of a probiotic preparation in improving the mammary gland health of a dairy cow. The application finds that a probiotic preparation including Akkermansia muciniphila can promote the proliferation of mammary gland epithelial cells of a dairy cow, inhibit the apoptosis thereof (alleviate inflammation of the mammary gland and promote the expression of milk secretion-related genes), and the like, which is helpful to maintain the mammary gland health of the dairy cow, and thus achieves the purpose of improving the milk yield and milk quality of the dairy cow.

[0006] In order to achieve the above-mentioned purpose, the technical scheme of the application is as follows:

[0007] An application of a probiotic preparation in improving the mammary gland health of a dairy cow, the probiotic preparation including: Akkermansia muciniphila and glycerol.

[0008] In the above technical scheme, the number of Akkermansia muciniphila in the probiotic preparation is greater than or equal to 5x10 9 CFU / mL.

[0009] In the above technical scheme, the Akkermansia muciniphila is Akkermansia muciniphila BAA-835 strain.

[0010] In the above technical scheme, the Akkermansia muciniphila BAA-835 strain is purchased from Hebei Beina Biotechnology Co., Ltd.

[0011] In the above technical scheme, the glycerol is glycerol with a volume fraction of 30%.

[0012] In the above technical scheme, the probiotic preparation of the application is prepared by the following method:

[0013] Akkermansia muciniphila is cultured by using BHI medium and 4% rumen fluid, and the culture is centrifuged, the supernatant is discarded, the bacterial slurry is washed with sterile physiological saline, the bacterial slurry is resuspended in glycerol, and the probiotic preparation is obtained.

[0014] In the above technical scheme, the culture condition is 85% N2, 5% CO2, 10% H2, and anaerobic culture at 37℃.

[0015] In the above technical scheme, the centrifugation is 8000g centrifugation at 4℃ for 5min.

[0016] In the above technical scheme, the bacterial slurry is washed with sterile physiological saline for 3 times.

[0017] In the above technical solution, the bacterial slurry is resuspended in glycerol with a volume fraction of 30%, and the number of Akkermansia muciniphila in the finally obtained probiotic preparation is ≥ 5 x 10 9 CFU / mL.

[0018] The beneficial effects of the present application are:

[0019] The present application finds that the microbial preparation made of Akkermansia muciniphila can help maintain the health of the mammary glands of dairy cows by promoting the proliferation of mammary epithelial cells, inhibiting their apoptosis (relieving inflammation of the mammary glands and promoting the expression of lactation-related genes), thereby improving the milk yield and quality of dairy cows. The preparation of the present application is safer, more efficient and non-polluting, and is non-toxic and harmless, which does not have a negative impact on the health of dairy cows. In addition, the probiotic bacteria are easy to obtain and easy to culture in large quantities, and the operation is simple, the raw materials are easy to obtain, and the method is simple, which can reduce the cost of manpower and material resources. BRIEF DESCRIPTION OF DRAWINGS

[0020] The present application will be further described in detail below in combination with the drawings and specific embodiments.

[0021] Figure 1 Figure 1 shows the effect of the probiotic preparation of the present application on the function of mammary gland cells of dairy cows. DETAILED DESCRIPTION

[0022] The present application provides an application of a probiotic preparation in improving the health of the mammary glands of dairy cows, which comprises: Akkermansia muciniphila, and glycerol. Preferably, the number of Akkermansia muciniphila in the probiotic preparation is ≥ 5 x 10 9 CFU / mL, and the Akkermansia muciniphila is Akkermansia muciniphila BAA-835 strain, which is purchased from Hebei Beina Biological Technology Co., Ltd.

[0023] The probiotic preparation of the present application can be prepared by the following preparation method:

[0024] Akkermansia muciniphila BAA-835 (BNCC 341917) bacteria (purchased from Hebei Beina Biotechnology Co., Ltd.) were cultured in BHI medium (+ 4% rumen fluid) at 85% N2, 5% CO2, 10% H2, 37°C anaerobic culture; the bacterial liquid with OD value of 0.8-1 was centrifuged at 8000g for 5min at 4°C, the supernatant was discarded, the bacterial slurry was washed with sterile physiological saline for 3 times, and the bacterial slurry was resuspended in glycerol with a volume fraction of 30%(v / v), and the number of Akkermansia muciniphila BAA-835 in the finally obtained probiotic preparation was ≥5×10 9 CFU / mL. Alternatively, the obtained probiotic preparation can be stored in a-80°C refrigerator.

[0025] The application of the probiotic preparation of the application in improving the health of the mammary glands of dairy cows is used for the feeding method of dairy cows, and the prepared probiotic preparation is fed for 90 days, 5mL per cow per day.

[0026] The application finds that the probiotic preparation containing Akkermansia muciniphila can promote the proliferation of mammary epithelial cells of dairy cows, inhibit apoptosis (alleviate inflammation of the mammary glands and promote the expression of lactation-related genes), etc., which is helpful to maintain the health of the mammary glands of dairy cows, and thus the purpose of improving the milk yield and milk quality of dairy cows is achieved.

[0027] Example

[0028] Akkermansia muciniphila BAA-835 (BNCC 341917) bacteria (purchased from Hebei Beina Biotechnology Co., Ltd.) were cultured in BHI medium (+ 4% rumen fluid) at 85% N2, 5% CO2, 10% H2, 37°C anaerobic culture; the bacterial liquid with OD value of 0.8-1 was centrifuged at 8000g for 5min at 4°C, the supernatant was discarded, the bacterial slurry was washed with sterile physiological saline for 3 times, and the bacterial slurry was resuspended in glycerol with a volume fraction of 30%(v / v), and the number of Akkermansia muciniphila BAA-835 in the finally obtained probiotic preparation was ≥5×10 9 CFU / mL. And the obtained probiotic preparation can be stored in a-80°C refrigerator.

[0029] The probiotic preparation prepared in the example is used for feeding dairy cows, and the feeding is carried out for 90 days, 5mL per cow per day, and the conditions are recorded as follows:

[0030] Feeding began in July 2022 and ended in September 2022. 78 dairy cows on the farm were fed the diet, while 52 cows served as a control group. Milk production and mammary gland health (indicators related to mammary gland inflammation) were measured monthly during the feeding period (Table 1). Simultaneously, 37 cows with high somatic cell counts were fed the diet to observe its effect on reducing somatic cell counts (Table 2). The effects of probiotic preparations on mammary cell function were observed using an in vitro mammary cell culture model. Figure 1 The effects of myxotroph-Ackermania on lactation performance and mammary gland health in dairy cows were assessed (Table 2).

[0031] Table 1. Effects of probiotic preparations on lactation and mammary gland health in dairy cows

[0032]

[0033] Table 1 shows that feeding dairy cows probiotic preparations significantly increased milk production in the second and third months of feeding. Simultaneously, the probiotic preparations significantly reduced the incidence of mastitis in dairy cows. However, they had no significant effect on milk fat percentage and milk protein content. This indicates that probiotic preparations can improve milk production and mammary gland health in dairy cows.

[0034] Table 2. Effects of probiotic preparations on somatic cell count in dairy cows

[0035]

[0036]

[0037] As shown in Table 2, the somatic cell count of dairy cows decreased significantly after one month of feeding with probiotic preparations, indicating that probiotic preparations can treat subclinical mastitis in dairy cows.

[0038] Figure 1 This is a diagram illustrating the effect of the probiotic preparation from the example on the function of bovine mammary gland cells. Figure 1 It can be seen that probiotic preparations can significantly promote mammary cell proliferation (significant increase in the proliferation-related gene CCND1) and inhibit cell apoptosis (significant increase in anti-apoptotic genes and significant decrease in pro-apoptotic genes). These results demonstrate at the cellular level that probiotic preparations have a significant beneficial regulatory effect on the mammary health of dairy cows.

[0039] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. Use of probiotics in the preparation of preparations for improving the health of dairy cow mammary glands, the probiotics comprising: Akkermansia muciniphila and glycerol; the Akkermansia muciniphila is Akkermansia muciniphila BAA-835 strain; the glycerol is glycerol with a volume fraction of 30%; the probiotics are prepared by the following method: the Akkermansia muciniphila is cultured in BHI medium and 4% rumen fluid until the OD value of the bacterial solution is 0.8-1, centrifuged, the supernatant is discarded, the bacterial slurry is washed with sterile normal saline, and the bacterial slurry is resuspended in glycerol to obtain the probiotics. The Akkermansia muciniphila BAA-835 strain is purchased from Hebei Beina Biotechnology Co., Ltd. the number of mucinophilic-Akkermansia in the probiotic is ≥ 5 x 10 9 CFU / mL; The culture conditions are: 85% N2, 5% CO2, 10% H2, and anaerobic culture at 37°C. The centrifugation is at 8000g for 5 min at 4°C. The bacterial slurry is washed with sterile normal saline for 3 times. The bacterial slurry is washed with sterile normal saline for 3 times.

2. Use according to claim 1, characterized in that, ​ 3. Use according to claim 1, characterized in that, ​ 4. Use according to claim 1, characterized in that, ​ 5. The use according to claim 1, characterized in that, ​

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