阅读说明：本技术 硝酸钙在缓解湖羊酸中毒中的应用 (Application of calcium nitrate in relieving Hu sheep acidosis ) 是由 郑琛 刘婷 范慧玉 刘绘汇 孙康 于 2021-08-25 设计创作，主要内容包括：本发明提供了硝酸盐在缓解湖羊酸中毒中的应用。本发明在饲料中添加硝酸钙喂养湖羊提高瘤胃液乙酸/总酸比例和氨态氮的含量,且处理与时间表现出显著的交互作用；硝酸钙可使湖羊瘤胃液pH维持在较高水平,缓解湖羊的酸中毒。(The invention provides application of nitrate in relieving Hu sheep acidosis. According to the invention, calcium nitrate is added into the feed to feed the Hu sheep to improve the acetic acid/total acid ratio of rumen fluid and the content of ammoniacal nitrogen, and the treatment and the time show obvious interaction; the calcium nitrate can maintain the pH of the Hu sheep rumen fluid at a higher level and relieve the acid poisoning of the Hu sheep.)

1. Application of nitrate in relieving acid poisoning of Hu sheep is provided.

2. The use according to claim 1, wherein the nitrate is present in an amount of 1-3% of the feed.

3. The application of the feed as claimed in claim 1, wherein the feed comprises 15-18 parts of corn straw, 9-11 parts of soybean hull, 1-3 parts of sunflower hull, 45-50 parts of corn, 10-12 parts of soybean meal, 2-4 parts of sesame cake, 3-6 parts of molasses, 0.2-0.5 part of stone powder, 0.60-0.8 part of NaCl, 0.5-1.50 parts of premix and 1-3.5 parts of nitrate by mass.

4. Use according to claim 3, characterized in that the premix provides 25mg of iron, 40mg of manganese, 40mg of zinc, 8mg of copper, 0.3mg of iodine, 0.2mg of selenium, 0.1mg of cobalt, 940IU of vitamin A, 111IU of vitamin D, 20IU of vitamin E per kg of feed.

5. The use according to claim 1, characterized in that the feed comprises, by mass, 17.76 parts of corn stalks, 10.34 parts of soybean hulls, 2.00 parts of sunflower hulls, 48.60 parts of corns, 11.35 parts of soybean meal, 3.00 parts of sesame cakes, 5.00 parts of molasses, 0.25 part of stone powder, 0.70 part of NaCl, 1.00 part of premix and 3 parts of nitrate.

6. Use according to claim 1, characterized in that the nitrate is calcium nitrate.

Technical Field

The invention belongs to the technical field of livestock breeding, and particularly relates to application of calcium nitrate in relieving Hu sheep acidosis.

Background

In the prior art, nitrate can be used as a source of non-protein nitrogen, and can change the internal environment of the gastrointestinal tract of ruminants and serum indexes. Homeostasis of the rumen environment in ruminants is critical to their own health. After the animals eat the feed containing nitrate, the nitrate is firstly reduced into nitrite in the rumen under the action of microorganisms, and finally ammoniacal nitrogen is generated. In this process, the reduction of nitrate requires the consumption of hydrogen, and since the participation of hydrogen is also required in the synthesis of propionic acid, the reduction of hydrogen inhibits the production of propionic acid and promotes the production of acetic acid. In addition, the reduction of hydrogen also helps to alleviate the inhibitory effect on fiber-degrading bacteria. Nitrite, an intermediate product of nitrate metabolism, has strong oxidation capacity and can reduce antioxidant factors and oxidation resistance of organisms in organisms. However, most of the current researches on nitrate are concentrated on the aspect of methane emission reduction, and the researches on the influence of the dynamic fermentation of the Hu sheep rumen and the serum antioxidant index are less. Therefore, calcium nitrate is added into the feed to study the influence of the calcium nitrate on the dynamic fermentation of the rumen of the Hu sheep and the oxidation resistance of serum.

In order to improve the production efficiency of ruminants, the feed ratio is generally improved in the fattening process of cattle and sheep, but the feed ratio also increases the risk of rumen acidosis, so that the rumen acidosis is a common nutritional metabolic disease in cattle and sheep farms at present. At present, the common method is to add alkaline substances such as baking soda and the like into the feed of cattle and sheep, but the nutritive value of the feed cannot be improved. On the basis of not increasing the feeding cost and manpower and material resources, the calcium nitrate is added into the Hu sheep feed, so that the effect of improving the crude protein level of the feed is achieved, the pH of the rumen of the Hu sheep is effectively improved, and acidosis is relieved, and the method is a very effective means.

Disclosure of Invention

In order to solve the technical problems, the invention provides an application of nitrate in relieving the acid poisoning of Hu sheep.

In order to achieve the purpose, the invention adopts the following technical scheme that:

application of nitrate in relieving acid poisoning of Hu sheep is provided.

In the application, the nitrate is preferably used in an amount of 1-3% of the feed.

The application is preferably that the feed comprises 15-18 parts of corn straws, 9-11 parts of soybean hulls, 1-3 parts of sunflower hulls, 45-50 parts of corns, 10-12 parts of soybean meal, 2-4 parts of sesame cakes, 3-6 parts of molasses, 0.2-0.5 part of mountain flour, 0.60-0.8 part of NaCl, 0.5-1.50 parts of premix and 1-3.5 parts of nitrate in parts by mass.

For use as described above, the premix preferably provides 25mg of iron (Fe), 40mg of manganese (Mn), 40mg of zinc (Zn), 8mg of copper (Cu), 0.3mg of iodine (I), 0.2mg of selenium (Se), 0.1mg of cobalt (Co), 940IU of Vitamin A (VA), 111IU of Vitamin D (VD), and 20IU of Vitamin E (VE) per kg of feed.

The feed comprises, by mass, 17.76 parts of corn straws, 10.34 parts of soybean hulls, 2.00 parts of sunflower hulls, 48.60 parts of corns, 11.35 parts of soybean meal, 3.00 parts of sesame cakes, 5.00 parts of molasses, 0.25 part of stone powder, 0.70 part of NaCl, 1.00 part of premix and 3 parts of nitrate.

For the above-mentioned applications, preferably, the nitrate is calcium nitrate.

The invention has the beneficial effects that:

the invention provides an application of calcium nitrate in relieving Hu sheep acidosis, wherein the calcium nitrate is added into feed to improve the acetic acid/total acid ratio of Hu sheep rumen fluid and the content of ammoniacal nitrogen, and the treatment and the time show obvious interaction; the calcium nitrate not only improves the nitrogen source supply for synthesizing the microbial mycoprotein of the rumen of the Hu sheep, but also improves the digestion and absorption of the rumen fibrous substance of the Hu sheep by promoting the proliferation of the fiber degrading bacteria, effectively improves the nutrient utilization rate, effectively maintains the pH value of the rumen fluid of the Hu sheep at a higher level, and relieves the acidosis of the Hu sheep.

Drawings

FIG. 1 shows the 24h dynamic change of the pH value of the rumen of a Hu sheep.

Detailed Description

The following examples are intended to further illustrate the invention but should not be construed as limiting it. Modifications and substitutions may be made thereto without departing from the spirit and scope of the invention.

Unless otherwise indicated, the technical means used in the examples are conventional means well known to those skilled in the art, and unless otherwise specified, the reagents used in the methods are analytically pure or above.

Example 1

6 Hu sheep with permanent rumen fistula were selected and randomly divided into 2 groups of 3 animals, one repeat each. The control group was fed basal diet and the test group was fed diet supplemented with 3% calcium nitrate. The test is carried out for 30 days in 2 stages, each stage is 15d, the transition stage (changing the feed and enabling the Hu sheep to adapt to the test conditions and the feed) is 10d, the positive test stage (formally starting to collect samples) is 5d, and the Hu sheep eat freely. The test is completed in the drought region environment of the cold region of the Chinese academy of sciences and the Satland test station of the engineering research institute. Before selecting test sheep, the sheep house used in the test is disinfected in all directions by using powerful disinfectant, and the disinfected sheep house allows disinfectant in the sheep house to volatilize for a period of time and then enters the sheep so as to avoid discomfort of the sheep. After the test formally begins, the test is fed in a single cage, and the test is fed with the feed once every morning and afternoon, so that the test can be fed freely and can drink water freely. The composition (by mass parts) and the nutrient components of the feed are shown in a table 1-1, and the feed is produced and processed by Gansu Runmu bioengineering Limited liability company.

TABLE 1-1 feed composition and its nutrient composition table (Dry matter basis)

Wherein, the premix provides the following components for each kg of feed: 25mg of iron (Fe), 40mg of manganese (Mn), 40mg of zinc (Zn), 8mg of copper (Cu), 0.3mg of iodine (I), 0.2mg of selenium (Se), 0.1mg of cobalt (Co), 940IU of Vitamin A (VA), 111IU of Vitamin D (VD), and 20IU of Vitamin E (VE).

Firstly, rumen fluid collection: rumen fluid 0h, 1h, 3h, 5h and 7h after meal is collected at the 4 th and 5d of the positive test period. The rumen chyme of the test sheep is taken out by a beaker after a fistula plug is removed, then the rumen chyme is filtered by 4 layers of gauze, and the filtered rumen fluid is filled into a 15mL centrifuge tube and then immediately stored at-80 ℃ for the measurement of Volatile Fatty Acid (VFA) and ammonia nitrogen.

1. Determination of VFA in rumen fluid: and (3) centrifuging rumen fluid at 5000rpm for 10min, taking 1mL of supernatant, putting the supernatant into a 1.5mL centrifuge tube, adding 0.2mL of 25% metaphosphoric acid solution (containing an internal standard substance of 2-ethyl butyric acid), fully mixing uniformly, putting the mixture into an ice water bath for more than 30min, centrifuging the mixture at 10000rpm for 10min, taking out the centrifuge tube, transferring the supernatant into a gas phase vial by using a liquid transfer gun, and then loading the vial on a machine for determination. VFA was determined using an Agilent 6890N gas chromatograph and an HP19091N-213 chromatographic column. The chromatographic analysis conditions were: the injection port temperature is 220 ℃, the split ratio is 40:1, and the nitrogen flow is 2.0 mL/min. The detector temperature was 250 ℃, the air flow rate was 450mL/min, the nitrogen flow rate was 45mL/min, and the hydrogen flow rate was 40 mL/min. Temperature programming: the initial temperature is 120 deg.C for 3min, and then the temperature is raised to 180 deg.C at 10 deg.C/min and kept for 1 min. The amount of the sample was 0.6. mu.L.

2. Determination of ammoniacal nitrogen in rumen fluid: the influence of the level of supplementary feed NDF on the development of digestive tracts of suckling lambs [ D ]. Lanzhou, Gansu agricultural university, 2013. the content of ammoniacal nitrogen in rumen fluid is determined by a medium method. After the rumen fluid is centrifuged at 5000rpm for 10min, 1mL of supernatant is transferred into a 10mL centrifuge tube, 4mL of 0.2M hydrochloric acid is added, and the mixture is shaken up. Then, 0.2mL of the mixture was aspirated, the mixture was placed in a 5mL centrifuge tube, 1mL of solution A (100 mL of a 14% sodium salicylate solution containing 0.08g of sodium nitroferricyanide) and 1mL of solution B (100 mL of a 0.3M NaOH solution containing 2mL of a sodium hypochlorite solution) were sequentially added, the mixture was thoroughly shaken, and the mixture was allowed to stand for 10 minutes. Then, 200. mu.L of the mixed solution was transferred to a 96-well plate, and the absorbance at a wavelength of 700nm was measured using a microplate reader. And calculating the ammonia nitrogen content in the rumen fluid according to the ammonia nitrogen standard curve.

The results of measuring the concentrations of VFA and ammoniacal nitrogen in the rumen fluid of Hu sheep are shown in tables 1-2. The measured data are all sorted by Excel, and SPSS 22.0 software is used for carrying out univariate variance analysis on the data by a general linear mixed model, wherein P is less than or equal to 0.05 to represent that the difference between groups is significant, and 0.05< P <0.1 to represent that the difference between groups has a significant trend.

TABLE 1-2 Effect of feed additions of calcium nitrate on VFA and ammoniacal nitrogen concentrations in Hu sheep rumen fluid

As can be seen from tables 1-2, the addition of calcium nitrate to the feed significantly increased the acetic acid/total acid ratio and the ammoniacal nitrogen concentration in the rumen fluid of Hu sheep (P < 0.05); the propionic acid/total acid ratio was significantly reduced (P < 0.05). The content of total acid in rumen fluid 1h after eating by the Hu sheep is obviously higher than 0h and 7h (P is less than 0.05); the ratio of propionic acid/total acid after eating is significantly higher than 0h, and the treatment produces significant interaction with time (P < 0.05); the acetic acid/propionic acid ratio after meal was significantly below 0h and treatment produced significant interaction with time (P < 0.05). The treatment and the time produce obvious interaction (P < 0.05) on the ammonia nitrogen concentration of the Hu sheep rumen fluid. In addition, calcium nitrate has a tendency to increase the acetic acid/propionic acid ratio of the hu-sheep rumen fluid (P ═ 0.068). Nitrate, as a class of nitrogenous compounds, provides a source of nitrogen for fermentation by ruminal microorganisms in ruminants, inhibits propionic acid production by consuming hydrogen during the production of ammoniacal nitrogen via nitrite in the rumen, and contributes to the proliferation of fiber-degrading bacteria. Therefore, the calcium nitrate not only improves the nitrogen source supply for synthesizing microbial mycoprotein, but also improves the digestion and absorption of the rumen fibrous substance of the Hu sheep by promoting the proliferation of the fiber degrading bacteria, thereby achieving the effect of improving the nutrient utilization rate.

3. The influence of different NFC/NDF ratio diets on the dynamic change of the rumen pH of the milk goats [ J ]. Chinese livestock raising journal, 2009,45(3):19-22. And (3) dynamically monitoring the pH value of the rumen of the test sheep for 24 hours by adopting a dynamic pH monitoring system 2d before the positive test period of each test period. At each successive dynamic pH monitoring, the electrodes were calibrated with pH 4.00 and pH 6.86 standard solutions and the calibration values recorded. After the electrode is calibrated, the electrode is inserted into the rumen through the rumen fistula and then the plug is plugged. The electrode is connected with the signal input end of a pH transmitter outside the fistula through a lead, and the signal output end of the pH transmitter is connected with the signal input end of a recorder.

The test uses 3 channels to display in real time, the pH value of 6 test sheep in each period is dynamically changed and divided into 2 days to finish, and the pH value is recorded for 1 time every 1 min. And then uploading the recorded data to a computer through a U disk special for a dynamic pH monitoring system, calling out the data in the U disk by using DTM data management software, and storing the data in an Excel document for analysis and processing. Determination of dynamic pH in rumen: the pH data collected from each sheep per day were pooled and the mean, maximum, minimum, and duration of pH below 5.60 and 5.80 and the area of the curve were calculated. The area of the curve is the product of the cumulative sum of the absolute values of the deviations below the pH threshold and the pH threshold, and the time interval, and is expressed in pH min/d. And the average pH value per hour was calculated for plotting a 24h dynamic pH change, the results of which are shown in table 2 and fig. 1.

TABLE 2 influence of calcium nitrate addition to diets on the dynamic pH of the rumen of Hu sheep

As can be seen from table 2, the addition of calcium nitrate to the diet significantly reduced the minimum value of the hu sheep rumen dynamic pH (P < 0.05), but had no significant effect on other dynamic pH values (P > 0.05), but the average pH was elevated. As can be seen from Table 2 and FIG. 1, calcium nitrate maintained the rumen pH of the Hu sheep at a higher level and for a shorter period of time than the pH of 5.60 and 5.80 than the control Hu sheep (P > 0.05). Rumen pH is an important indicator of rumen fermentation of ruminants, whether the rumen pH is within a normal range or not directly affects the effect of rumen fermentation, rumen pH is too low to facilitate the growth of cellulolytic bacteria, affect the digestion and absorption of nutrients, and rumen can damage epithelial cells of rumen when being in a low pH environment for a long time. In this test, the rumen pH decreased and then increased after feeding the control group hu sheep, while the rumen pH increased and then decreased after feeding the calcium nitrate group hu sheep, and the ruminant rumen pH of 5.2 was generally regarded as the critical point for acute acidosis, and 5.6 and 5.8 as the critical points for subacute acidosis. In the invention, the calcium nitrate has a reducing effect on the maintaining time and the curve area of the Hu sheep rumen fluid pH lower than 5.80 and 5.60, and the average value of the Hu sheep rumen pH is maintained at a higher level, which shows that the calcium nitrate has the effect of relieving Hu sheep acidosis.