中国的奶牛养殖业已经步入了规模性养殖，奶业的健康平稳发展，对调整居民膳食纤维、满足青少年蛋白质需求、带动相关产业发展等都具有重要的意义。微量元素是动物生长发育过程中不可缺少的，它们摄入不足、过量、比例不平衡均会不同程度影响机体正常的生理功能，甚至导致疾病的发生。奶牛肢蹄病是奶牛生产中一种常见的病，主要原因是日粮中营养的不平衡，轻则引起跛行，严重者可致瘫痪，增加了饲养成本。因此，高效而合理地补充微量元素，是提高奶牛生产性能、减少疾病发生、降低饲养成本、促进行业稳定发展的重要措施。奶牛饲养者一般普遍采用在饲料中加入微量元素添加剂的方式来补充微量元素，但消化道的环境对微量元素的吸收有较大的影响，导致生产实践中经常出现饲喂不足或饲喂过度的现象。因此探索出简单的有效的微量元素补充途径十分必要。注射剂的特点主要是作用迅速、剂量准确，能够避免有效成分受消化道环境的影响，因此可以考虑使用注射法给奶牛补充微量元素。在仔猪、兔、山羊等动物上使用微量元素注射液的研究有很多，但在奶牛方面设计、制备并运用复合微量元素注射液尚未有报道。本试验根据NRC奶牛饲养标准提出的奶牛对各种微量元素的需要量，设计并制备复合微量元素注射液，在优化生产工艺和检测稳定性后，研究其对泌乳期奶牛的产奶量、乳品质、抗氧化能力、血清微量元素含量和奶牛蹄病等方面的影响，从而为复合微量元素注射液在奶牛养殖中的应用提供试验依据。

试验一：复合微量元素注射液制备工艺研究

根据NRC奶牛营养标准中的奶牛对铜、铁、钴、锰、硒、锌的绝对需要量，选择二水合氯化铜 （分子式CuCl2·2H2O）、七水合硫酸亚铁（分子式FeSO4·7H2O）、七水合硫酸钴（分子式CoSO4·7H2O）、四水合氯化锰（分子式MnCl2·4H2O）、氯化锌（分子式ZnCl2）、山梨醇（分子式C6H14O6）作为原料，使用注射用水溶解。制备工艺为在清洁环境下先用1L注射用水将原料溶解，加入1%体积分数活性炭吸附，先后使用滤纸和0.22 μm滤器过滤，121℃高压灭菌15分钟，最终将注射液分装于棕色西林瓶中。对复合微量元素注射液的制备工艺进行优化，试验结果显示，在溶解水温60℃、活性炭添加量1%体积分数、pH调整为2.2时，注射液具有最佳的性状。对注射液进行质量检查，试验结果显示，澄明度检查、无菌检查均符合相关规定，在比格犬和山羊的股二头肌进行注射也未观察到明显的刺激性。对复合微量元素注射液的有效成分进行检测，得出各有效成分的含量分别为Cu：522.18 μg/ml、Fe：876.93 μg/ml、Mn：127.03 μg/ml、Zn：1118.72 μg/ml、Co：133.17 μg/ml、Se：62.15 μg/ml。

试验二：复合微量元素注射液稳定性研究

根据《兽药稳定性实验研究指导原则》，本试验采用光照影响因素试验、温度加速试验、长期试验、经典恒温加速试验来研究复合微量元素注射液的稳定性。光照影响因素试验结果显示，将复合微量元素注射液开口放于照度4500±500 lx的环境中，注射液的外观没有改变，但pH值改变较大，具体表现为pH降低，各元素的含量也有一定量的衰减，因此受光照影响较大，要选择避光条件储存；温度加速试验结果显示，复合微量元素注射液在温度40±2 ℃、相对湿度75±5 %的环境中放置6个月，注射液的性状未发生明显的改变，pH有所降低，各元素含量有所衰减，但变化都很轻微；长期试验结果显示，将复合微量元素注射液室温避光保存于室温环境中，每隔三个月检测一次，药品在性状、pH、含量方面都能保持相对稳定；通过经典恒温加速试验计算出复合微量元素注射液的有效期为18个月。

试验三：复合微量元素注射液对奶牛的生产性能、血液指标和蹄病的影响

本试验通过对奶牛注射复合微量元素注射液，评价其对奶牛生产性能、血液指标和蹄病影响。根据五分制步态评分标准，试验选取年龄、胎次、泌乳量相近的奶牛16头，随机分为对照组和试验组，每组8头，其中每组包含4头肢蹄患病牛和4头健康牛。分别在第0 d、7 d、14 d给试验组奶牛肌肉注射20 ml复合微量元素注射液，对照组奶牛肌肉注射20 ml生理盐水。每隔15 d采集血液样本，每隔30 d采集乳样，检测相关指标，并观察奶牛的蹄病和跛行情况。试验结果显示：与对照组相比，试验组奶牛30 d时产奶量从31.34 kg增长到36.6 kg，显著高于同期对照组的32.26 kg（p < 0.05）；体细胞数相较于对照组也有显著下降（p < 0.05）；试验组奶牛60 d乳脂率为4.19 %，显著高于对照组的乳脂率3.25 %（p < 0.05）；乳蛋白率、奶牛体况评分、尿素氮则未发现明显差异（p > 0.05）。抗氧化指标方面，与对照组相比，试验组奶牛血清中的GSH-PX含量极显著增加（ p < 0.01），SOD显著增加（p < 0.05），T-AOC和MDA有略微升高趋势，但差异不显著（p > 0.05）。试验组血清中铜、锰、锌、硒、钴、铁的含量显著增加（p < 0.05）。试验组蹄病和跛行的发病率在60d时均低于对照组。本试验表明，复合微量元素注射液能提高奶牛的产奶量，并且改善乳品质，一定程度上提高奶牛机体的抗氧化能力，提高血清中微量元素的含量，并且对预防和治疗蹄病有一定积极作用。

The dairy farming industry in China has entered large-scale farming. The healthy and steady development of the dairy industry is of great significance for adjusting the dietary fiber of residents, meeting the protein needs of young people, and driving the development of related industries. Trace elements are indispensable in the process of animal growth and development. Inadequate intake, excess, and imbalanced proportions will affect the normal physiological functions of the body to varying degrees, and even lead to the occurrence of diseases. Limb hoof disease is a common disease in dairy cow production. The main reason is the imbalance of nutrients in the diet, which can cause lameness, and can cause paralysis in severe cases, which increases the cost of feeding. Therefore, efficient and reasonable supplementation of trace elements is an important measure to improve the production performance of dairy cows, reduce the occurrence of diseases, reduce feeding costs, and promote the stable development of the industry. Dairy cattle breeders generally use trace element additives to supplement trace elements, but the digestive tract environment has a greater impact on the absorption of trace elements, resulting in insufficient or over-feeding in production practices. phenomenon. Therefore, it is necessary to explore a simple and effective way to supplement trace elements. The characteristics of the injection are mainly rapid action and accurate dosage, which can avoid the effective ingredients from being affected by the environment of the digestive tract. Therefore, it may be considered to use the injection method to supplement trace elements to the cows. There are many studies on the use of trace element injections in piglets, rabbits, goats and other animals, but the design, preparation and application of compound trace element injections in dairy cows have not been reported. In this experiment, according to the requirements of dairy cows for various trace elements proposed by the NRC dairy feeding standards, composite trace element injections were designed and prepared. After optimizing the production process and testing stability, the milk production and milk production of lactating cows were studied. The effects of quality, antioxidant capacity, serum trace element content, and dairy cow hoof disease provide an experimental basis for the application of compound trace element injection in dairy farming.

Experiment 1: Study on the preparation process of compound trace element injection

According to the absolute requirements of cows for copper, iron, cobalt, manganese, selenium, and zinc in the NRC dairy nutrition standards, choose copper chloride dihydrate (molecular formula CuCl2·2H2O), ferrous sulfate heptahydrate (molecular formula FeSO4·7H2O), Cobalt sulfate heptahydrate (molecular formula CoSO4·7H2O), manganese chloride tetrahydrate (molecular formula MnCl2·4H2O), zinc chloride (molecular formula ZnCl2), and sorbitol (molecular formula C6H14O6) are used as raw materials to dissolve with water for injection. The preparation process is to dissolve the raw materials with 1L of water for injection in a clean environment, add 1% volume fraction of activated carbon for adsorption, filter with filter paper and 0.22 μm filter, autoclave at 121°C for 15 minutes, and finally divide the injection solution into brown cillin In the bottle. The preparation process of the compound trace element injection was optimized. The test results showed that the injection had the best properties when the dissolved water temperature was 60°C, the activated carbon addition was 1% by volume, and the pH was adjusted to 2.2. The quality of the injection was checked. The experimental results showed that the clarity test and sterility test were in compliance with the relevant regulations. No obvious irritation was observed in the injection of the Biceps and goat femoral biceps. The effective components of the compound trace element injection were tested, and the contents of each effective component were Cu: 522.18 μg/ml, Fe: 876.93 μg/ml, Mn: 127.03 μg/ml, Zn: 1118.72 μg/ml, Co : 133.17 μg/ml, Se: 62.15 μg/ml.

Experiment 2: Study on the stability of compound trace element injection

According to "Guidelines for Experimental Research on Stability of Veterinary Drugs", this experiment uses light influencing factor experiment, temperature acceleration experiment, long-term experiment and classic constant temperature acceleration experiment to study the stability of compound trace element injection. The experimental results of light influencing factors showed that the appearance of the injection did not change when the opening of the compound trace element injection was placed in an environment with an illumination of 4500±500lx, but the pH value changed greatly, which was manifested by the decrease in pH and the content of each element. The amount of attenuation, so it is greatly affected by light, you should choose to store in dark conditions; temperature acceleration test results show that the compound trace element injection is placed in an environment with a temperature of 40±2℃ and a relative humidity of 75±5% for 6 months. There is no obvious change in the properties of the injection, the pH has decreased, and the content of each element has attenuated, but the changes are very slight; long-term experimental results show that the compound trace element injection is stored in a room temperature environment at room temperature and protected from light. Tested once every three months, the drug can remain relatively stable in terms of traits, pH, and content; the validity period of the compound trace element injection is calculated by the classic constant temperature accelerated test for 18 months.

Experiment 3: The effect of compound trace element injection on the performance of dairy cows, serum ions and hoof disease

In this experiment, the compound trace element injection was injected into the cows to evaluate their effects on the production performance, blood indexes and hoof disease of the cows. According to the five-point gait scoring standard, 16 cows with similar age, parity, and milk production were randomly divided into a control group and an experimental group, each group consisting of 8 cows. 4 healthy cattle. On the 0 d, 7 d and 14 d, the experimental group was intramuscularly injected with 20 ml of compound trace element injection, and the control group was intramuscularly injected with 20 ml of normal saline. Blood samples were collected every 15 days and milk samples were collected every 30 days to detect relevant indicators and observe the condition of hoof disease and lameness of the cows. The test results showed that: compared with the control group, the milk yield of the experimental group increased from 31.34 kg to 36.6 kg at 30 days, which was significantly higher than that of the control group at 32.26 kg (p<0.05); the number of somatic cells was also higher than that of the control group. Significantly decreased (p<0.05); the 60-day milk fat rate of the experimental group was 4.19%, which was significantly higher than that of the control group (3.25%) (p<0.05); no significant differences were found in milk protein rate, cow body condition score, and urea nitrogen (P>0.05). Compared with the control group, compared with the control group, the GSH-PX content in the serum of the experimental group was significantly increased (p<0.01), SOD was significantly increased (p<0.05), and T-AOC and MDA increased slightly. But the difference was not significant (p>0.05). The serum, copper, manganese, zinc, selenium, cobalt and iron contents in the experimental group increased significantly (p<0.05). The incidence of hoof disease and lameness in the experimental group was lower than that in the control group at 60 days. This test shows that the compound trace element injection can improve the milk production of dairy cows, and improve the milk quality, to a certain extent, improve the antioxidant capacity of the dairy cow body, increase the content of trace elements in the serum, and have certain effect on the prevention and treatment of hoof disease positive effects.

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