Application of walnut green peel extract in rumen fermentation regulation product preparation
Walnut husk extract, prepared by extracting active ingredients from the green husk of Xinjiang walnuts, solves the problem of rumen acidosis in ruminants, achieves efficient rumen fermentation regulation, and avoids the side effects of antibiotics.
Patent Information
- Application Number
- CN202310181072.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2019-07-23
- Publication Date
- 2026-02-10
- Estimated Expiration
- 2039-07-23
AI Technical Summary
There is a lack of effective antibiotic alternatives in the current technology to regulate rumen fermentation in ruminants, which leads to rumen acidosis. Furthermore, long-term use of antibiotics can cause bacterial resistance and drug residues, affecting food safety and health.
Walnut green husk extract was used as a rumen fermentation regulator. The active ingredients were extracted from Xinjiang walnut green husk through a specific process to prepare a dark brown viscous extract, which was added to sheep diets to regulate rumen pH and cellulase activity.
It significantly increases rumen pH, enhances cellulase activity, reduces propionic and valerate concentrations, prevents rumen acidosis, achieves efficient rumen fermentation regulation, and avoids the side effects of antibiotics.
Smart Images
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Abstract
Description
[0001] This application is a divisional application of a patent application entitled "A Novel Plant Rumen Fermentation Regulator and Its Application in Sheep Diets", the original application was filed on July 23, 2019, and the application number is 201910664431.0. Technical Field
[0002] This invention belongs to the field of feed preparation technology, and in particular relates to the application of a walnut green husk extract in the preparation of rumen fermentation regulation products. Background Technology
[0003] In pursuit of maximizing the economic benefits of cattle and sheep farming, increasing milk production, and accelerating fattening and slaughter, livestock producers are continuously increasing the amount of concentrate feed used in ruminant livestock. While the extensive use of high-concentrate feed has significantly improved the production performance of ruminants, it has also led to nutritional metabolic diseases such as rumen acidosis, laminitis, and forestomach atony. In particular, the excessive feeding of concentrate feed or the overuse of silage and distillers' grains causes rumen fermentation to proceed in a manner conducive to the production of large amounts of propionic acid, resulting in a rapid drop in rumen pH to below 5.5. This reduces the activity of fiber-degrading bacteria, causing severe digestive disorders and abnormal fermentation of stomach contents, ultimately leading to acidosis. Currently, rumen acidosis caused by this practice has become a significant factor restricting the improvement of production efficiency in dairy cattle, beef cattle, and sheep.
[0004] Currently, many farms add antibiotics such as monensin and virginiamycin to the diet to prevent rumen acidosis. While antibiotics can inhibit the activity of lactic acid bacteria and Streptococcus bovis and prevent excessive pH drops, long-term use of antibiotics can lead to problems such as bacterial resistance, drug residues, and even endanger food safety and human health. Therefore, finding antibiotic alternatives to improve rumen fermentation in ruminants is urgently needed.
[0005] Plant extracts are feed additives made from the whole plant or parts of one or more natural plants through physical extraction or bio-fermentation. They possess nutritional, growth-promoting, feed utilization-enhancing, and animal product quality-improving effects. Due to their antibacterial, antioxidant, and low-toxicity properties, plant extracts are frequently used as novel feed additives in ruminant production. Studies have shown that extracts from Ranunculaceae plants can significantly increase the pH of in vitro rumen fermentation broth, with a trend towards increased total flammable fat (TVFA), higher molar percentages of acetic and butyric acids, and a decreased molar percentage of propionic acid. Sow thistle extract can increase the pH of rumen fluid from in vitro wheat starch fermentation and can be used as a regulator of rumen fermentation. In addition, yucca extract, plant essential oils, hovenia tannins, and ginkgo leaf extract also have certain regulatory effects on rumen fermentation.
[0006] Walnut husk, also known as green dragon skin, is the green outer skin covering the walnut. It contains various active ingredients, including 45 volatile components, 27 phenols, and juglone, among which the main active ingredients are naphthoquinones, polyphenols, flavonoids, and terpenes. Studies have demonstrated that juglone, polyphenols, and flavonoids possess antibacterial and antioxidant effects. Therefore, its extracts have the potential to replace antibiotics, exhibiting green and safe characteristics, providing a theoretical basis for the research and application of this invention. Currently, walnuts are widely cultivated in Xinjiang, resulting in a very large yield; however, the development of walnut husk resources has not yet formed a mature technical system, indicating enormous potential for resource development. Summary of the Invention
[0007] To address the shortcomings of existing technologies, namely the lack of a specific technology for preparing rumen fermentation regulators using high-yield walnut husk extracts and applying them in sheep diets, this invention provides an application of walnut husk extract in the preparation of rumen fermentation regulator products, a rumen fermentation regulator product made from walnut husk extract, and a specific method for using the product.
[0008] This invention is achieved through the following technical solution:
[0009] This invention provides an application of walnut husk extract in the preparation of rumen fermentation regulation products. The walnut husk extract uses Xinjiang walnut husks as raw material. Fresh walnut husks are dried at 65°C, pulverized through a 100-mesh sieve, and then extracted with an organic extraction solvent at a material-to-liquid ratio of 1:10. In a bioreactor (115), extraction was carried out at 37°C and 900 r / min for 8 hours. Afterward, the extract was poured out and allowed to stand for 6 hours. The clarified supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 r / min to obtain a dark brown, viscous walnut husk extract. The rumen fermentation regulating product is any one or more of the following: a product that prevents a decrease in rumen pH, a product that increases the activity of rumen cellulase filter paper enzyme, and a product that increases the activity of rumen cellulase cellobiase.
[0010] Preferably, the walnut husk is the husk of Xinjiang walnuts.
[0011] Preferably, the extraction solvent is ethanol, methanol, or acetone.
[0012] Preferably, the extraction solvent methanol has a mass percentage concentration of 50% to 75%;
[0013] Preferably, the extraction solvent ethanol has a mass percentage concentration of 25% to 50%;
[0014] Preferably, the mass percentage concentration of the extraction solvent acetone is 25% to 75%.
[0015] The present invention also provides a rumen fermentation regulation product for the aforementioned applications.
[0016] The present invention also provides a method for using the rumen fermentation regulating product, wherein the rumen fermentation regulating product is added at 0.5% of the daily feed weight, mixed evenly, and then fed.
[0017] Preferably, the method of use is as follows: the daily feed contains 65 parts concentrate and 35 parts roughage, and the rumen fermentation regulating product is added at 0.5% of the daily feed.
[0018] In this invention, concentrates refer to common foods such as corn, wheat, and wheat bran; roughage refers to common foods such as corn stalks, cottonseed meal, and rapeseed meal.
[0019] Preferably, the rumen animal being fed is a sheep.
[0020] By implementing the specific content of this invention, the following beneficial effects can be achieved:
[0021] Adding 0.5% of the rumen fermentation regulating product of this application to a high-concentrate diet for sheep can prevent a decrease in rumen pH, increase the activity of rumen cellulase and acetic acid levels, and significantly reduce the concentrations of propionic acid and valerate. Furthermore, experimental results showed that after using the rumen fermentation regulating product of this application, the average rumen pH of sheep increased by 0.24 throughout the day, filter paper enzyme activity increased by 73%, cellobiase activity increased by 66%, acetic acid increased by 29%, and propionic acid and valerate decreased by 28% and 43%, respectively. This indicates that the product of this application can play a highly efficient role in regulating rumen fermentation during high-concentrate feeding in rumen animals, and can prevent rumen acidosis caused by high-concentrate feeding. Detailed Implementation
[0022] The present invention will now be illustrated with examples, but the present invention is not limited to the examples described below.
[0023] The materials used in this invention include: anhydrous ethanol, anhydrous methanol, acetone, aqueous ethanol solution, aqueous methanol solution, aqueous acetone solution, and distilled water. All reagents and materials can be purchased through public channels, and the equipment and instruments used in the process are all common in the field.
[0024] All materials, reagents and instruments used in this invention are well known in the art, but this does not limit the implementation of the invention. Other reagents and devices well known in the art can also be applied to the implementation of the following embodiments of the invention.
[0025] The following embodiments further illustrate the content of the present invention, but should not be construed as limiting the present invention. Any modifications or substitutions made to the methods, steps, or conditions of the present invention without departing from the spirit and essence of the invention are within the scope of the present invention.
[0026] Example 1: Preparation of Walnut Green Peel Extract
[0027] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 25% ethanol, 50% methanol, or 25% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0028] Example 2: Preparation of Walnut Green Peel Extract
[0029] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 25% ethanol, 65% methanol, or 50% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0030] Example 3: Preparation of Walnut Green Peel Extract
[0031] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 25% ethanol, 75% methanol, or 75% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0032] Example 4: Preparation of Walnut Green Peel Extract
[0033] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 35% ethanol, 50% methanol, or 25% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0034] Example 5: Preparation of Walnut Green Peel Extract
[0035] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 35% ethanol, 65% methanol, or 50% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0036] Example 6: Preparation of Walnut Green Peel Extract
[0037] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 35% ethanol, 75% methanol, or 75% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0038] Example 7: Preparation of Walnut Green Peel Extract
[0039] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ and then pulverized through a 100-mesh sieve. Using 50% ethanol, 50% methanol, or 25% acetone as the extraction solvent, the mixture is placed in an Eppendorf 115 bioreactor at a solid-liquid ratio of 1:10 and extracted at 37℃ and 900 rpm for 8 hours. Afterward, the extract is poured off and allowed to stand for 6 hours. The clear supernatant is filtered through four layers of gauze. The filtered extract is then transferred to a rotary evaporator and evaporated at 55℃ and 170 rpm to obtain a dark brown, viscous walnut green husk extract.
[0040] Example 8: Preparation of a novel plant rumen fermentation regulator
[0041] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 50% ethanol, 65% methanol, or 50% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0042] Example 9: Preparation of Walnut Green Peel Extract
[0043] After the walnuts mature, the green husks are removed, and the walnuts are naturally sun-dried at 65℃ before being crushed and passed through a 100-mesh sieve. An extraction solvent of 50% ethanol, 75% methanol, or 75% acetone is used, with a material-to-liquid ratio of 1:10. Extraction was carried out in a 115 bioreactor at 37°C and 900 rpm for 8 hours. Afterwards, the extract was poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 rpm to obtain a dark brown, viscous walnut husk extract.
[0044] Example 10: Application of walnut husk extract in sheep diets
[0045] Based on the walnut husk extracts provided in Examples 1 to 9 above, walnut husk extracts are added at 0.5% of the daily feed weight, mixed evenly, and then used to prepare sheep feed.
[0046] Example 11: Sheep diet containing walnut husk extract
[0047] This invention provides a novel plant rumen fermentation regulator for sheep diets that do not require antibiotics. The sheep diet contains 65 parts concentrate, 35 parts roughage, and 0.5% walnut husk extract by weight.
[0048] Example 12: Application of Rumen Fermentation Regulation Products Made from Walnut Green Peel Extract
[0049] Five healthy, non-pregnant ewes of the Small-tailed Han sheep breed, with an average weight of 40 kg and permanent rumen fistulas, were selected and housed in individual pens. Based on the walnut husk extracts provided in Examples 1 to 9, the sheep diet prepared in Example 11 was fed for 12 days. Rumen fluid was collected continuously for 3 days, 5 times a day: once before feeding (0 h), and once each at 1.5, 3, 6, and 10 h after feeding. The rumen fluid pH, digestive enzyme activity, and volatile fatty acids were measured. The results are shown in Table 1.
[0050] Table 1. Effects of adding plant rumen fermentation regulators on pH, enzyme activity, and volatile fatty acids in rumen fluid.
[0051]
[0052]
[0053] Table 1 shows that adding 0.5% walnut husk extract to a high-concentrate diet for sheep prevented a decrease in rumen pH, increasing the average rumen pH by 0.24 throughout the day. The activities of two rumen cellulases, filter paper enzyme and cellobiase, increased by 73% and 66%, respectively. Acetic acid levels increased by 29%, while propionic acid and valerate levels decreased by 28% and 43%, respectively. Therefore, adding 0.5% walnut husk extract to a high-concentrate diet for sheep can effectively regulate rumen fermentation during high-concentrate feeding and prevent rumen acidosis caused by high-concentrate diets.
[0054] As described above, the present invention can be well implemented. The above embodiments are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, all changes and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the present invention.
Claims
1. The application of walnut husk extract in the preparation of rumen fermentation-regulated products, characterized in that, The walnut green husk extract was obtained by the following preparation method: fresh walnut green husks were dried at 65°C, pulverized through a 100-mesh sieve, and extracted with an ethanol solution of 25%–35% by mass as the extraction solvent. The solution was placed in an Eppendorf 115 bioreactor at a material-to-liquid ratio of 1:10 and extracted at 37°C and 900 r / min for 8 hours. The extract was then poured out and allowed to stand for 6 hours. The clear supernatant was filtered through four layers of gauze. The filtered extract was then transferred to a rotary evaporator and evaporated at 55°C and 170 r / min to obtain a dark brown, viscous walnut green husk extract. The rumen fermentation regulating product is any one or more of the following: a product that increases the activity of rumen cellulase filter paper enzyme, a product that increases the activity of rumen cellulase cellobiase, or a product that prevents the decrease of rumen pH.
2. The application according to claim 1, characterized in that, The green husk of the walnut mentioned is the green husk of Xinjiang walnuts.