COMPOSITION FOR IMPROVING MILK QUALITY AND PREPARATION METHOD AND APPLICATION THEREOF

Abstract

The disclosure discloses a composition containing melatonin as an effective ingredient, and its application in improving DHI production performance and improving milk quality. The composition containing melatonin as an effective ingredient according to the present disclosure can significantly reduce the somatic cell count in milk and improve milk quality. Under the conditions of different seasons, ages, and lactation periods, the administration of the composition containing melatonin as an active ingredient according to the present disclosure can significantly reduce the somatic cell score of cow milk (p<0.01), increase the protein content and lactose content in milk, and meanwhile reduce the fat content and urea nitrogen content in milk. In summary, the composition containing melatonin as an effective ingredient according to the present disclosure can improve the quality of milk and increase the efficiency of dairy cow breeding.

Claims

1. A composition for improving milk quality of a dairy cow, comprising an active ingredient containing melatonin.

2. The composition according to claim 1, wherein the active ingredient of the composition is melatonin.

3. The composition according to claim 1, wherein the composition comprises melatonin at a concentration of 1 to 20 mg/mL.

4. The composition according to claim 1, further comprising absolute ethanol and physiological saline; and the volume ratio of absolute ethanol to physiological saline is in a range of (1-3):(1-2).

5. The composition according to claim 4, wherein the volume ratio of absolute ethanol to physiological saline is in a range of 3:2.

6. The composition according to claim 1, wherein the composition is in a form of an injection.

7. A method for preparing the composition of claim 1, comprising the following steps: dissolving melatonin in absolute ethanol and then adding physiological saline, wherein the steps are carried out in dark.

8. The method according to claim 7, wherein the active ingredient of the composition is melatonin.

9. The method according to claim 7, wherein the composition comprises melatonin at a concentration of 1 to 20 mg/mL.

10. The method according to claim 7, wherein the volume ratio of absolute ethanol to physiological saline is in a range of (1-3):(1-2).

11. The method according to claim 10, wherein the volume ratio of absolute ethanol to physiological saline is in a range of 3:2.

12. The method according to claim 7, wherein the composition is prepared as a form of injection.

13. A method for improving milk quality of a dairy cow, comprising a step of administering a composition comprising an active ingredient containing melatonin, to a dairy cow having a somatic cell count of 300,000/mL to 1,000,000/mL; wherein the step of administering comprises administering the composition by injection at a dosing of 2 to 3 mL.

14. The method according to claim 13, wherein the composition is administered by injection once a day for 3 to 6 consecutive days.

15. The method according to claim 13, wherein the improvement of the milk quality comprises one or more selected from a group consisting of: (1) reducing a somatic cell count in milk; (2) increasing a milk protein content; (3) increasing a lactose content; (4) increasing a dry matter content in milk; (5) reducing a fat content; and (6) reducing a urea nitrogen content.

16. The method according to claim 13, wherein the active ingredient of the composition is melatonin.

17. The method according to claim 13, wherein the composition comprises melatonin at a concentration of 1 to 20 mg/mL.

18. The method according to claim 13, wherein the composition further comprises absolute ethanol and physiological saline; the volume ratio of absolute ethanol to physiological saline is in a range of (1-3):(1-2).

19. The method according to claim 18, wherein the volume ratio of absolute ethanol to physiological saline is in a range of 3:2.

Description

DESCRIPTION OF THE DRAWINGS

[0047] FIG. 1 is a diagram showing the effect of the injection of melatonin on DHI indicators of the milk from dairy cows in different seasons.

[0048] FIG. 2 is a diagram showing the effect of the injection of melatonin on DHI indicators of the milk from dairy cows of different ages.

[0049] FIG. 3 is a diagram showing the effect of the injection of melatonin on DHI indicators of the milk from dairy cows of different lactation periods.

[0050] FIG. 4 is a diagram showing the analysis results of the effect of the injection of melatonin on DHI indicators.

DETAILED DESCRIPTION OF THE INVENTION

[0051] The following examples are merely used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0052] The reagents and materials used in the following examples are all commercially available products.

[0053] The purity of Melatonin is required to reach 99.9%.

Example 1. Preparation Process for the Injections of Melatonin

[0054] Materials: melatonin and absolute ethanol.

[0055] Preparation process: accurately weigh 464.54 mg of melatonin, dissolve it with absolute ethanol in a 100 mL volumetric flask and stir well, then add 0.9% of physiological saline and stir well, set a final volume of 100 mL (anhydrous ethanol:saline=3:2), and finally dispense into 2 mL injection dose. The whole process is done in a dark room.

Example 2. Analysis of the DHI Indicators of Milk after Subcutaneous Injection of Melatonin

[0056] 1. Experimental Process

[0057] (1) According to the monthly DHI test reports from the dairy farm, Holstein cows having somatic cell counts of 300,000/mL to 1,000,000/mL are selected to be fed separately, and are ensured to have no significant difference in physical health. Season groups: spring n=90, summer n=74, autumn n=83, winter n=81 (number of cows). Age groups: 2-year-old n=23; 3-year-old n=80; 4-year-old n=66; 5-year-old n=56; 6-year-old n=27; 7-year-old n=19 (number of cows). Lactation period groups: peak-lactation n=15, mid-lactation n=38, end-lactation n=59 (number of cows) (initial-lactation: days 1-15 after calving; peak-lactation: days 16-100 after calving; mid-lactation: days 101-200 after calving; end-lactation: day 201 after calving to weaning).

[0058] (2) The dairy cows having high somatic cell counts are injected with 2 mL melatonin solution every day for 4 consecutive days in each month. 15 days after the injection, fresh milk was collected and stored in 50 mL centrifuge tubes, and was tested for the changes in somatic cell counts. The cows were fed with total mixed ration, and were fed three times at 06:00, 12:00, and 18:00 respectively. Then, the cows entered the sports ground to move freely, drink water and rest.

[0059] (3) The sampling bottles were numbered and added with 2-3 drops of potassium dichromate saturated solution, and were used for collecting the milk samples from the milk by the first milking of the cows. The milk sample for each cow was in a volume of 40 mL and placed in a centrifuge tube. After sampling, all the remaining milk was drained from the tubes which were cleaned to avoid contamination between the samples. The samples were stored at 2-7° C. The instrument for measuring the DHI indicators is Milkscan FT+&FC automatic milk compositional analyzer and somatic cell counter. The measurement was carried out in the National Laboratory for Preparation of Standard Materials of DHI.

[0060] 2. Experimental Results

[0061] (1) The somatic cell scores of the milk were significantly reduced in all seasons, with the greatest reduction in summer (FIG. 1A). The protein content was significantly increased (p<0.05) by injecting melatonin in autumn (3.51%±0.05 VS 3.64%±0.05) (FIG. 1C). The lactose content was increased after injecting melatonin, and was increased significantly in spring (4.89%±0.02 VS 4.94%±0.02), summer (4.96%±0.03 VS 5.06%±0.02) and autumn (4.99%±0.03 VS 5.07%±0.04) (p<0.05) (FIG. 1D). The fat content was decreased after injecting melatonin, and was decreased significantly in spring (5.18%±0.12 VS 4.78%±0.14) (p<0.05) (FIG. 1E).

[0062] The results are shown in FIG. 1, in which: [0063] A. changes in somatic cell scores: [0064] spring (5.96±0.08 VS 4.89±0.15), [0065] summer (6.06±0.12 VS 4.09±0.20), [0066] autumn (6.03±0.11 VS 4.55±0.16), and [0067] winter (6.12±0.11 VS 4.34±0.17); [0068] B. changes in milk production: [0069] spring (32.08±1.04 VS 32.11±1.15, kg/d), [0070] summer (34.09±1.06 VS 32.69±1.00, kg/d), [0071] autumn (32.88±1.02 VS 32.07±1.05, kg/d), and [0072] winter (31.67±1.11 VS 31.82±1.17, kg/d); [0073] C. changes in protein contents: [0074] spring (3.46%±0.04 VS 3.49%±0.04), [0075] summer (3.45%±0.06 VS 3.45%±0.04), [0076] autumn (3.51%±0.05 VS 3.64%±0.05), and [0077] winter (3.39%±0.05 VS 3.43%±0.05); [0078] D. changes in lactose contents: [0079] spring (4.89%±0.02 VS 4.94%±0.02), [0080] summer (4.96%±0.03 VS 5.06%±0.02), [0081] autumn (4.99%±0.03 VS 5.07%±0.04), and [0082] winter (4.70%±0.07 VS 4.84%±0.04); [0083] E. changes in fat contents: [0084] spring (5.18%±0.12 VS 4.78%±0.14), [0085] summer (4.48%±0.16 VS 4.20%±0.12), [0086] autumn (4.80%±0.11 VS 4.68%±0.13), and [0087] winter (4.82%±0.10 VS 4.75%±0.11); [0088] F. changes in dry matter contents: [0089] spring (14.23%±0.14 VS 13.92%±0.15), [0090] summer (13.61%±0.17 VS 13.42%±0.13), [0091] autumn (14.00%±0.13 VS 14.10%±0.14), and [0092] winter (13.68%±0.10 VS 13.65%±0.12); [0093] G. changes in urea nitrogen contents: [0094] spring (13.08±0.39 VS 14.35±0.45, mg/dL), [0095] summer (16.59±0.64 VS 14.19±0.50, mg/dL), [0096] autumn (12.14±0.43 VS 11.72±0.36, mg/dL), and [0097] winter (17.39±0.47 VS 15.94±0.51, mg/dL);

[0098] wherein, spring n=90, summer n=74, autumn n=83, and winter n=81 (unit: number of cows).

[0099] (2) Injection of melatonin reduced the somatic cell scores of the milk from Holstein cows at different ages, and had significant effect in Holstein cows of 2-, 3-, 4-, 5- and 7-year-old (p<0.05) (FIG. 2A). The lactose content was increased by injecting melatonin, and was significantly increased in cows of 3- (4.95%±0.03 VS 5.03%±0.02) and 4-year-old (4.80%±0.04 VS 4.94%±0.03) (p<0.05) (FIG. 2D). The fat content was reduced by injecting melatonin, and was significantly reduced in cows of 3-year-old (p<0.05) (FIG. 2E). The dry matter content of the milk was significantly reduced by injecting melatonin in Holstein cows of 3-year-old (13.82%±0.12 VS 13.57%±0.13) (FIG. 2F).

[0100] The results are shown in FIG. 2, in which: [0101] A. changes in somatic cell scores: [0102] 2-year-old (5.30±0.27 VS 3.87±0.33), [0103] 3-year-old (5.70±0.15 VS 4.31±0.18), [0104] 4-year-old (5.89±0.17 VS 4.68±0.18), [0105] 5-year-old (5.50±0.18 VS 4.04±0.19), [0106] 6-year-old (5.63±0.26 VS 5.19±0.27), and [0107] 7-year-old (5.91±0.20 VS 5.03±0.29); [0108] B. changes in milk production: [0109] 2-year-old (32.06±0.95 VS 32.12±1.63, kg/d), [0110] 3-year-old (31.72±1.04 VS 32.11±0.96, kg/d), [0111] 4-year-old (30.85±1.19 VS 30.03±1.22, kg/d), [0112] 5-year-old (35.03±1.54 VS 33.56±1.52, kg/d), [0113] 6-year-old (33.71±2.45 VS 35.19±2.67, kg/d), and [0114] 7-year-old (33.54±1.73 VS 30.34±2.08, kg/d); [0115] C. changes in protein contents: [0116] 2-year-old (3.37%±0.07 VS 3.41%±0.09), [0117] 3-year-old (3.43%±0.04 VS 3.44%±0.04), [0118] 4-year-old (3.57%±0.06 VS 3.66%±0.06), [0119] 5-year-old (3.44%±0.06 VS 3.55%±0.07), [0120] 6-year-old (3.38%±0.07 VS 3.38%±0.07), and [0121] 7-year-old (3.20%±0.06 VS 3.32%±0.07); [0122] D. changes in lactose contents: [0123] 2-year-old (5.20%±0.04 VS 5.19%±0.03), [0124] 3-year-old (4.95%±0.03 VS 5.03%±0.02), [0125] 4-year-old (4.80%±0.04 VS 4.94%±0.03), [0126] 5-year-old (4.80%±0.04 VS 4.86%±0.05), [0127] 6-year-old (4.85%±0.05 VS 4.91%±0.06), and [0128] 7-year-old (4.90%±0.06 VS 4.93%±0.06); [0129] E. changes in fat contents: [0130] 2-year-old (4.65%±0.24 VS 4.54%±0.26), [0131] 3-year-old (4.73%±0.12 VS 4.36%±0.11), [0132] 4-year-old (4.97%±0.16 VS 4.69%±0.13), [0133] 5-year-old (4.83%±0.14 VS 4.62%±0.16), [0134] 6-year-old (5.14%±0.18 VS 5.27%±0.21), and [0135] 7-year-old (5.16%±0.37 VS 4.66%±0.37); [0136] F. changes in dry matter contents: [0137] 2-year-old (13.91%±0.22 VS 13.87%±0.27), [0138] 3-year-old (13.82%±0.12 VS 13.57%±0.13), [0139] 4-year-old (14.19%±0.17 VS 14.18%±0.15), [0140] 5-year-old (13.82%±0.17 VS 13.87%±0.18), [0141] 6-year-old (14.14%±0.20 VS 14.13%±0.24), and [0142] 7-year-old (13.80%±0.36 VS 13.48%±0.32); [0143] G. changes in urea nitrogen contents: [0144] 2-year-old (12.78±0.90 VS 13.90±0.80, mg/dL), [0145] 3-year-old (14.09±0.53 VS 13.58±0.46, mg/dL), [0146] 4-year-old (14.30±0.61 VS 13.32±0.49, mg/dL), [0147] 5-year-old (14.95±0.52 VS 14.09±0.59, mg/dL), [0148] 6-year-old (15.38±0.90 VS 15.79±0.94, mg/dL), and [0149] 7-year-old (16.47±1.64 VS 15.11±1.34, mg/dL).

[0150] All data are expressed as mean±S.E.; the symbol “*” represents that there is significant differences between different groups; 2-year-old n=23; 3-year-old n=80; 4-year-old n=66; 5-year-old n=56; 6-year-old n=27; and 7-year-old n=19 (unit: number of cows).

[0151] (3) The somatic cell scores were significantly reduced by the injection of melatonin in different lactation stages, and were significantly reduced in mid-lactation (3.92±0.25 VS 2.00±0.17) and end-lactation (31.58±1.22 VS 31.25±1.29, kg/d) (p<0.05) (FIG. 3A). The lactose contents were increased by the injection of melatonin, and were significantly increased in peak-lactation (3.47%±0.06 VS 3.35%±0.06) and end-lactation (3.49%±0.04 VS 3.40%±0.04) (FIG. 3D).

[0152] The results are shown in FIG. 3, in which: [0153] A. changes in somatic cell scores: [0154] peak-lactation (2.86±0.32 VS 2.04±0.24), [0155] mid-lactation (3.92±0.25 VS 2.00±0.17), and [0156] end-lactation (4.34±0.16 VS 1.99±0.15); [0157] B. changes in milk production: [0158] peak-lactation (32.93±2.34 VS 38.80±2.06, kg/d), [0159] mid-lactation (39.98±1.35 VS 39.08±1.79, kg/d), and [0160] end-lactation (31.58±1.22 VS 31.25±1.29, kg/d); [0161] C. changes in protein contents: [0162] peak-lactation (3.47%±0.06 VS 3.35%±0.06), [0163] mid-lactation (3.45%±0.04 VS 3.45%±0.05), and [0164] end-lactation (3.49%±0.04 VS 3.40%±0.04); [0165] D. changes in lactose contents: [0166] peak-lactation (5.15±0.05% VS 5.25%±0.04), [0167] mid-lactation (5.14%±0.03 VS 5.19%±0.02), and [0168] end-lactation (5.12%±0.03 VS 5.20%±0.02); [0169] E. changes in fat contents: [0170] peak-lactation (4.45%±0.19 VS 3.87%±0.26), [0171] mid-lactation (4.20%±0.12 VS 4.22%±0.11), and [0172] end-lactation (4.07%±0.08 VS 4.18%±0.12); [0173] F. changes in dry matter contents: [0174] peak-lactation (13.76%±0.16 VS 13.17%±0.29), [0175] mid-lactation (13.49%±0.14 VS 13.56%±0.14), and [0176] end-lactation (13.38%±0.09 VS 13.47%±0.13); [0177] G. changes in urea nitrogen contents: [0178] peak-lactation (11.93±0.65 VS 11.12±0.64, mg/dL), [0179] mid-lactation (12.67±0.65 VS 12.11±0.54, mg/dL), and [0180] end-lactation (11.45±0.38 VS 12.04±0.38, mg/dL);

[0181] wherein, peak-lactation n=15, mid-lactation n=38, end-lactation n=59 (unit: number of cows).

[0182] (4) The changes in the DHI indicators of Holstein cows were recorded for a whole year and were used to analyze the influence of melatonin on various DHI indicators. With the injection of melatonin, the somatic cell score was significantly (p<0.01) reduced by 63.54%, the fat content of milk was significantly (p<0.05) reduced by 11.31%, and the urea nitrogen content of milk was significantly (p<0.05) reduced by 11.08%. With the injection of melatonin, the protein content of milk was significantly (p<0.05) increased by 3.79%, and the lactose content was significantly (p<0.05) increased to 4.74%, and the daily milk production and dry matter content were also be influenced to a certain extent.

[0183] The results are shown in FIG. 4.

[0184] As shown by FIG. 4, the DHI indicators including the somatic cell score, lactose content, urea nitrogen content, protein content and fat content were influenced significantly (p<0.05), in which the somatic cell score was most significantly affected (p<0.01), n=328 (unit: number of cows).

[0185] The present disclosure deeply analyzed the changes in the DHI indicators of milk after injecting exogenous melatonin, and reveals the mechanism in which melatonin improves the milk quality. The experimental results of the present disclosure demonstrate that the somatic cell counts in the milk of cows can be significantly reduced by injecting the dairy cows with 4.64 mg of melatonin for 4 consecutive days. It is also found that melatonin can reduce the concentration of cortisol, the number of white blood cells, lymphocytes and neutrophils; increase the concentrations of albumin, alanine aminotransferase and serum lactate dehydrogenase. The above research shows that the injection of melatonin can reduce the somatic cell counts in the milk of cows and increase the immunity of cows.

[0186] The present disclosure unexpectedly finds that it is simple to prepare an injection having melatonin as the main active ingredient, and the somatic cell counts in cow milk can be significantly reduced by injecting the melatonin injection into cows having somatic cell counts of 300,000/mL to 1,000,000/mL in different seasons, ages or lactation periods. This further shows that the melatonin injection according to the present disclosure can significantly reduce the somatic cell count in cow milk. At the same time, it is found by detecting the milk quality that the melatonin injection can not only increase the protein and dry matter contents in the milk, but also increase the lactose content in the milk, with reducing the fat and urea nitrogen contents in the milk. Thus, the melatonin injection of the present disclosure has more remarkable and excellent effects on improving milk quality.

[0187] The above examples are only used to describe the preferred embodiments of the present disclosure, but not intended to limit the scope of the present disclosure. Without departing from the spirit of the present disclosure, any variation and improvement made by those skilled person in the art based on the technical solutions of the present disclosure shall all fall within the protection scope defined by the attached claims of the present disclosure.