畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (12): 2016-2021.doi: 10.11843/j.issn.0366-6964.2013.12.023

• 临床兽医 • 上一篇    下一篇

金黄色葡萄球菌外毒素及荚膜特异性卵黄抗体联合治疗奶牛乳房炎效果研究

王林会1,李晓宇1,金礼吉1,2,李淑英2,徐永平1,2*   

  1. (1. 大连理工大学生命科学与技术学院,大连 116024;2. 动物性食品安全保障技术教育部工程研究中心,大连 116020)
  • 收稿日期:2013-06-13 出版日期:2013-12-23 发布日期:2013-12-23
  • 通讯作者: 徐永平,教授,E-mail: yongpingxu.dut@gmail.com
  • 作者简介:王林会(1985-),男,吉林长春人,博士生,主要从事奶牛乳房炎研究,E-mail: wanglinhui1985@aliyun.com
  • 基金资助:

    中央高校基本科研业务费(DUT13JB04);国家自然科学基金(30371053;31001053)

Efficacy of a Combination of Egg Yolk Immunoglobulins against Staphylococcus aureus Exotoxins and Capsules in the Treatment of Bovine Mastitis

WANG Lin-hui1, LI Xiao-yu1, JIN Li-ji1,2, LI Shu-ying2, XU Yong-ping1,2*   

  1. (1. School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China; 2. MOE (Ministry of Education) Center for Food Safety of Animal Origin, Dalian 116020, China)
  • Received:2013-06-13 Online:2013-12-23 Published:2013-12-23

摘要:

本研究目的是评价金黄色葡萄球菌外毒素特异性卵黄抗体(IgY-toxins)及58336型金黄色葡萄球菌荚膜特异性卵黄抗体(IgY-T5IgY-T8IgY-T336)联合使用对奶牛乳房炎的治疗效果。乳酸脱氢酶试验表明,IgY-toxins能够有效中和外毒素对乳腺上皮细胞MAC-T的破坏作用。荧光显微镜观察发现,5 mg·mL-1IgY-T336可有效阻断金黄色葡萄球菌RN6390MAC-T的侵袭。与单独使用IgY-T336相比,5 mg·mL-1IgY-T3360.5 mg·mL-1IgY-toxins联合使用能够更加有效地保护MAC-T细胞,同时显著改善细胞的生长形态。临床型乳房炎试验表明,与PBS对照相比,对金黄色葡萄球菌感染乳池每天注射各20 mgIgY-T5IgY-T8IgY-T336IgY-toxins可显著降低(P<0.001)病牛乳汁的体细胞数(SCC),20 mgIgY-T5IgY-T8IgY-T336或者20 mgIgY-toxins虽然也可降低体细胞数,但是降低幅度显著低于联合治疗组(P<0.01)。所有治疗组奶牛乳汁的凝块水平都显著低于对照组(P<0.05)。本研究表明,荚膜及外毒素特异性卵黄抗体联合使用比其单独使用对乳房炎的治疗效果更好,在应用卵黄抗体治疗乳房炎时应该将荚膜和外毒素都列为抗体的靶点。

Abstract:

The objective of this study was to evaluate the efficacy of a combination of egg yolk immunoglobulins (IgYs) specific for exotoxins (IgY-toxins) and serotypes 5, 8 and 336 capsules of Staphylococcus aureus (IgY-T5, IgY-T8, IgY-T336, respectively) in the treatment of bovine mastitis. A lactate dehydrogenase assay indicated that IgY-toxins prevented mammary epithelial cells (MAC-T) from lysing. An internalization assay demonstrated that 5 mg·mL-1 IgY-T336 markedly blocked the invasion of a GFP-expressing S. aureus RN6390 to MAC-T cells, and a combination of 5 mg·mL-1 of IgY-T336 and 0.5 mg·mL-1 of IgY-toxins provided the cells with greater protection and improved growth morphology compared with the IgY-T336 treatment alone. A field trial for clinical mastitis indicated that two doses of a combination of 20 mg each of IgY-T5, IgY-T8 and IgY-T336 plus 20 mg of IgY-toxins daily for six days significantly reduced somatic cell counts (SCC) compared with PBS control (P < 0.001). Treatment with IgY-T5, IgY-T8 and IgY-T336 or IgY-toxins alone produced a slight reduction in SCC compared with PBS control. However, the reduction was much lower than that in IgY combination treatment (P < 0.01). Both surface capsules and secreted exotoxins should be targeted in the treatment of S. aureus-caused bovine mastitis with IgY.

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