血链素对三种念珠菌菌体细胞膜通透性及超微结构影响的研究

发布时间：2018-06-25 05:56

本文选题：血链素 + 念珠菌　；参考：《佳木斯大学》2014年硕士论文

【摘要】：目的：研究血链素（Streptococcus sanguis bacteriocin）分别作用白色念珠菌(Candida albicans, C.albicans)、热带念珠菌（Candida tropicalis, C.tropicalis)及光滑念珠菌（Candida glabrata，C. glabrata）：1.探讨血链素的抑菌作用特点；2.检测三种念珠菌菌体细胞内小分子无机磷和大分子物质泄漏情况；3.观察血链素分别作用三种念珠菌后，菌体细胞超微结构的变化。方法： 1.血链素的提取：将血链球菌标准株ATCC10556复苏并厌氧培养，，通过超速低温离心、反复冻融、超声破碎等方法提取血链球菌菌体细胞内有效的抑菌物质，即血链素。 2.采用液体培养抑菌法和纸片扩散法，观察血链素分别对白色念珠菌、热带念珠菌及光滑念珠菌的抑制作用特点。 3.通过钼酸铵比色法及紫外吸收物质法,分别检测血链素对白色念珠菌、热带念珠菌及光滑念珠菌菌体细胞内磷外泄量及紫外吸收物质的渗透率； 4.利用扫描电镜(scanning electron microscopy, SEM)和透射电镜(transmissionelectron microscope，TEM)观察白色念珠菌、热带念珠菌及光滑念珠菌经血链素作用后的菌体细胞超微结构变化。结果： 1.血链素分别作用于白色念珠菌、热带念珠菌及光滑念珠菌后，其抑菌环直径均㧐7mm。且两两念珠菌之间的抑菌动力学特点存在相似性，血链素分别对三种念珠菌的抑制作用均在24h达高峰。 2.经血链素作用的白色念珠菌、热带念珠菌及光滑念珠菌菌体细胞内小分子无机磷和大分子物质发生泄漏。 3.通过SEM观察：血链素对白色念珠菌、热带念珠菌及光滑念珠菌作用24h时菌体细胞表面出现缺损、凹陷以及大小不一的突起，菌体呈现不规则形态。通过TEM观察：血链素对白色念珠菌作用24h时，菌体细胞表面粗糙、皱缩、形态不规则，细胞壁及细胞膜厚度变薄、结构分界不清，细胞核消失，胞质不均匀；血链素对热带念珠菌作用24h时，细胞壁及细胞膜厚度变薄，菌体细胞出现质壁分离，细胞壁及细胞膜连续性断裂或破坏，细胞质固缩，细胞核溶解；血链素对光滑念珠菌作用24h时，菌体细胞膜及细胞壁呈现凹坑状缺损，细胞壁及细胞膜厚度变薄，细胞核消失，出现空泡状。结论： 1.血链素分别对白色念珠菌、热带念珠菌及光滑念珠菌均有显著抑制作用以及相似的抑菌动力学特点。 2.血链素可使白色念珠菌、热带念珠菌及光滑念珠菌菌体细胞膜的通透性发生改变。 3.血链素可导致白色念珠菌、热带念珠菌及光滑念珠菌菌体细胞的超微结构发生改变。
[Abstract]:Objective: to study the effects of Streptococcus sanguis bacteriocin) on Candida albicans (C. albicans), C. tropicalis (C. tropicalis) and C. glabrata (Candida glabrata C. glabrata). Objective: to study the bacteriostatic characteristics of blood chain hormone. To detect the leakage of inorganic phosphorus and macromolecular substances in the cells of three Candida species. To observe the ultrastructure of three Candida species treated with streptavidin. Methods: 1. Extraction of streptavidin: the standard strain ATCC10556 was resuscitated and anaerobically cultured. Streptococcus sanguis strain ATCC10556 was extracted by ultracentrifugation, repeated freezing and thawing, ultrasonic crushing and so on. The inhibitory effects of streptavidin on Candida albicans, Candida tropicalis and Candida smoothing were observed by liquid culture and disk diffusion respectively. 3. By means of ammonium molybdate colorimetry and ultraviolet absorption method, the amount of phosphorus in the cells of Candida albicans, Candida tropicalis and Candida smoothing cells and the permeability of ultraviolet absorbents were determined respectively. 4. The ultrastructure of Candida albicans, Candida tropicalis and Candida smooth were observed by scanning electron microscope (scanning electron microscopy,) and transmission electron microscopy (transmissionelectron). Results: 1. The diameter of the inhibitory ring was 7 mm after the blood chain was treated with Candida albicans, Candida tropicalis and Candida smooth, respectively. The inhibitory kinetic characteristics of Candida albicans were similar, and the inhibitory effects of streptavidin on the three Candida species reached the peak at 24 h. 2. The leakage of small molecule inorganic phosphorus and macromolecular substance in the cells of Candida albicans, Candida tropicalis and Candida smooth cells was observed. SEM observation showed that the cell surface of Candida albicans, Candida tropicalis and Candida smooth exposed to streptavidin for 24 hours had defect, depression and different size protrusions, and the cells showed irregular morphology. Tem observation showed that the cell surface was rough, crumpled, irregular, the thickness of cell wall and cell membrane was thinning, the structure boundary was unclear, the nucleus disappeared and the cytoplasm was not uniform after 24 hours of action of hemchain hormone on Candida albicans. The thickness of cell wall and cell membrane became thinner, the cell wall and cell membrane were separated, the continuity of cell wall and cell membrane was broken or destroyed, the cytoplasm shrank and the nucleus dissolved. The cell membrane and cell wall of Candida smoothing exposed to strepsin for 24 hours showed pit defect, cell wall and cell membrane thickness thinned, cell nucleus disappeared and vacuolated. Conclusion: 1. The blood chain factor had significant inhibitory effect on Candida albicans, Candida tropicalis and Candida smooth, respectively. 2. The membrane permeability of Candida albicans, Candida tropicalis and Candida smooth could be changed by streptavidin. The ultrastructure of Candida albicans, Candida tropicalis and Candida smooth cells was changed by streptavidin.
【学位授予单位】：佳木斯大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781.54

【参考文献】