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1、石河子大学硕士学位论文奶牛乳头保护膜的研制及临床试验研究姓名：蔡曰鹏申请学位级别：硕士专业：临床兽医学指导教师：孙延鸣20100601I摘 要 奶牛乳房炎是牛场中最常见的疾病之一，对奶业生产造成巨大的经济损失。本病主要由多种非特定病原微生物感染或各种损伤引起。奶牛乳房炎的治疗首选的是抗生素疗 法，由于其产生的抗生素残留、耐药菌株等问题，危害到消费者的身体健康，并给预防和控制该病带来了困难，因此对奶牛乳房炎的防治要以“预防为主、治疗为辅”的方针，因此开发一种安全、无毒、无药残、使用方便，能有效预防乳房炎的制剂就显得尤为重要。本试验通过在干奶期使用乳头保护膜预防乳房炎为切入点，研制了奶牛乳头保护膜
2、并进行临床试验。 1 保护膜溶液各组分的优化组合 用壳聚糖和聚乙烯醇- 1 2 4 为主要成膜材料，加入相应的增塑剂、增强剂，本试验在单因素试验的基础上，选出最合适的成膜材料和相应的增塑剂、增强剂，并确定各因素的用量范围，最终通过正交试验设计法，以膜拉长倍数、感官柔韧性、膜附着性、成膜时间、外观质量效果为判定标准，筛选出保护膜各成份的最优水平组合。结果表明，最合适的成膜材料为聚乙烯醇- 124，增塑剂为甘油，增强剂为 CMC，其用量分别为：聚乙烯醇- 124 质量百分浓度为 8.0%、甘油的体积分数 3.0%、CMC 质量浓度为 0.05%。 2 皮肤刺激性和毒性试验 为了验证保护膜是否对皮肤
3、具有刺激性和毒性，以健康白兔为试验动物，背部脱毛，涂上奶牛乳头保护膜溶液，观察试验兔背部涂膜区有无红斑和水肿等明显的变化；全身表现、有无死亡情况、采食情况、呼吸、四肢活动等的变化。结果显示试验兔背部涂膜区未出现红斑、水肿等变化；未出现死亡及不良状况，采食、呼吸、四肢活动均未出现明显异常。 表明该乳头保护膜液对皮肤无刺激性、无毒性，可以用于干奶牛的临床试验。 3 留样观察试验 为了研究奶牛乳头保护膜溶液的稳定性，将制备好的保护膜溶液进行性能测定后，密封保存，在室内温度条件下，分别存放在阳光可以直射到的地方和避光的地方，记录保护膜溶液的外观情况，并每 3 个月对膜的性能进行测定。结果表明，存放于阳
4、光可以直射到的地方的保护膜溶液，在第 1 5 个月时，外观颜色稍微变白，其它性能未见变化，pH 值介于 6.51- 6.92；避光存放的保护膜溶液， 在 15 个月时，其外观没有肉眼可见变化，其它性能也未见变化，pH 值介于 6.53- 6.93。 4 奶牛乳头保护膜的临床试验 在干奶期使用乳头保护膜，对奶牛临床型和隐性乳房炎的预防效果观察。选取 3 2 4头干奶牛，1 2 9 6 个乳区，将每头奶牛 4 个乳区分为左右两部分，左侧使用奶牛乳头保护膜，做为试验组；右侧不做任何处理，做为对照组。以奶牛离开产房第一次回奶厅挤奶为期限，用 L M T 法检测隐性乳房炎；在奶牛产后 2 个月内，根据乳
5、房的外观症状、触诊情况、乳汁变化检测临床型乳房炎。试验组乳区临床型乳房炎乳区阳性率为 1 . 0 8 % ，对照组乳区阳性率 1 7 . 4 4 % （P 0 . 0 1 ） ；试验组乳区隐性乳房炎乳区阳性率为 1 0 . 3 1 % ，对照II组乳区阳性率 4 6 . 4 5 % （P 0 . 0 1 ） 。表明在干奶期使用奶牛乳头保护膜对临床型和隐性乳房炎具有较好的预防效果。 关键词：奶牛；乳房炎；乳头保护膜；临床试验； IIIAbstract Mastitis is one of the common and multiple disease in cow campus.It can c
6、ause massy economy loss to dairy production and potenia hazardous to the user health at the same time. Many kinds of non- definite causative organism infection and various kinds injury cause this disease. The antibiotic therapy generated many problems,such as antibiotics resid and drug resistant str
7、ain.These problems will be harmful to consumer health and make troubles to the prophylaxis and control for the mastitis treatment.So it is especially important to develop a kind of praepara tum which is safe, asepsis, no- drug leftover, convenient and effective. Mastitis prevention and cure was majo
8、rly taken the policy of therapy for primary and prophylaxis to assist.In this experiment,we take using the nipple protective membrane to prevent the mastitis in the dry milk stage as the incision point.We also develop nipple protective membrane and run clinical test. 1. Preparation of cownipple prot
9、ective membrane The chitosan and polyvinyl alcohol were used as the main film- former, and the propotional plasticizer and intensifier were added. On the base of the single factor experiment, the suitable film- former and the propotional plasticizer and intensifier were selected, and the dosage rang
10、e of each factor was decided. By the experiment of orthogonal design, the membrane extendere multiple, the sensory flexibility, the membrane adhesion, the membrane formation time and the appearance effects were regared as the assessed standard, and the best combination of every composition in the pr
11、otective membrane solution were selected. The results showed that the suitable film- former was polyvinyl alcohol with the mass concentration was 80g/mL, and the plasticizer wasglycerol with the volume fraction was 3%, and theintensifier was CMC with the mass concentration was 0.5g/mL. 2.The trial o
12、f skin stimulation test In order to verify the stimulation whether the protective membrane had to the skin , the healthy rabbits were choosed as experimental animal, the hair were shaved and the protective membrane solution was overlayed on the back skin. Then we observed the film coating district w
13、hether had the red spot or hydrops changes, finally no changesemerged. The appearance all over the body and the death by poisoning, including red swelling degree of the epilate district skin, fur gloss, the changes of eyes , mucosa, foraging, breathing andextremities activity and so on. It showed th
14、at the nipple protective membrane stimulation on the skin is free, and could use clinical experiment in the cow dry period. 3. Remaining sample and observation experiment In order to study and research the store time of the dairy cow nipple protective membrane solution,we put solution on the windows
15、ill and protection from light place indoors after IVperformance measure.We also held it airtightness on room temperature.Then we record appearance of the solution and tested performance every three month.The result show that the performance of the solution putting on the windowsill indoors has no ch
16、ange except appearance color turn slightly white on the eighteen month. The hydrogenion concentration(pH) was 6.51 to 6.92 and had no change. The performance of the solution putting protected from light has no change on the eighteen month. The hydrogenion concentration(pH) was 6.53to 6.93 and had no change.It show that we can store the solution at least eighteen month on room temperature without deteriorative change.So we can use it conveniently in production and practice. 4 .clinical
