地霉菌的毛发穿孔试验研究和扫描电镜观察

发布时间：2018-06-06 16:20

本文选题：林生地霉 + 皮损株　；参考：《河北医科大学》2010年硕士论文

【摘要】： 目的:地霉菌(Geotrichum)为酵母样真菌,属半知菌类、丝孢纲、丝孢目、丛梗孢科,其代表菌种白地霉(Geotrichum candidum ),是一种条件致病菌,多引起支气管感染,偶可致全身播散性感染。林生地霉(Geotircum sivicola)是地霉属中新发现的一个种,2002年首次从巴西果蝇和印度柞蚕幼虫身上分离得到。同年,我科首次从一脓癣患儿皮损中分离得到一株林生地霉。2005年,我科又自一例中毒性表皮坏死松解症患者的血液中分离出一株林生地霉。李秀丽等从形态学、营养生理学、分子生物学等各个角度对林生地霉进行初步研究,并且通过皮肤感染的动物实验及毛发受侵试验对其致病力进行了初步探讨。本研究通过对不同年龄组毛发进行改良毛发穿孔实验,一方面在光镜和电镜下观察林生地霉血液株,林生地霉皮损株和白地霉的菌丝及孢子的形态及其对毛发破坏的程度,为临床上诊断地霉所致头发的感染提供指导。另一方面比较其对不同年龄组毛发感染时间的差异,进而评价地霉菌对毛发的侵袭力。方法:林生地霉系我科保存的经中科院微生物研究所鉴定菌种,白地霉购自中科院微生物研究所。将林生地霉血液株、林生地霉皮损株和白地霉分别接种于改良沙堡氏培养基上,在37℃温箱培养14天,挑取菌落置于生理盐水中,经振荡分别制备成林生地霉皮损株、林生地霉血液株及白地霉菌悬液,并用血细胞计数板将菌悬液中孢子数控制在108/ ml～109/ ml范围备用。取4个年龄组的毛发(小于2岁、2～13岁、13～19岁、大于19岁)分别置于沙氏葡萄糖蛋白胨琼脂培养基,然后在各年龄组毛发上分别接种林生地霉皮损株、林生地霉血液株及白地霉菌悬液3滴,置于370温箱培养,并取各年龄组毛发数根,不加菌悬液作为空白对照。在接种后2、4、6、7、8、9、10、11周各取数根毛发分别用10 % KOH和酚棉蓝染色压片光镜下观察菌丝和大分生孢子的形态,毛发有破坏者进一步应用日立S23500N扫描电镜摄片,以观察林生地霉皮损株、林生地霉血液株和白地霉形态学上的差异以及受感染毛发的结构变化。结果:比较林生地霉血液株、林生地霉皮损株和白地霉对不同年龄组毛发感染时间的差异,以毛发上有菌丝或孢子附着且毛发有实质损伤为穿孔标准。林生地霉和白地霉均可致毛发破坏。在每年龄组中,不同菌株间毛发穿孔时间比较差异有显著性(P0.05),另外无论是皮损株,血液株还是白地霉感染毛发所需的时间随着年龄的增长,也随之延长,经统计学处理有显著性差异(P0.05)。对不同菌株间的差异进行两两比较,发现在各年龄组中,林生地霉血液株、皮损株和白地霉的两两比较差异均有显著性(P0.05),其中林生地霉血液株的毛发穿孔时间范围在53-64天,平均为58.07±3.15天;林生地霉皮损株的毛发穿孔时间范围在57-66天,平均为61.05±2.55天;白地霉的毛发穿孔时间范围在61-74天,平均为67.11±3.78天。提示林生地霉血液株导致毛发感染的时间最短,白地霉导致毛发感染的时间最长,因此林生地霉血液株对毛发的破坏性较其他两种菌株强。对不同年龄组间的差异进行两两比较,发现除小于2岁组与2～13岁组之间比较差异无显著性外(P0.05),其余各年龄组的两两比较差异均有显著性,提示年龄越小,毛发越易被感染。无论是林生地霉还是白地霉,在光镜下均可观察到毛发边缘出现毛刷样改变,破坏严重时毛发表面可出现锲形缺损,或毛发边缘凹凸不平。光镜可见较多的矩形、椭圆形关节孢子和圆形小分生孢子,丰富的芽生孢子,少量的厚膜孢子、菌丝稀少。在电镜下观察,林生地霉对毛发破坏的程度较白地霉严重,且林生地霉血液株较林生地霉皮损株对毛发破坏的程度严重。白地霉的菌丝仅附着在毛发表面并侵入毛小皮的缝隙间,使毛小皮排列稀松、少量脱落,从而造成皮质的轻度缺损;而林生地霉皮损株的菌丝严重侵蚀皮质,使毛小皮大量的破坏和脱落,造成毛发的片状缺损,毛干变细;林生地霉血液株的菌丝还可造成毛发的劈开或断裂,严重者造成毛发髓质的破坏。结论: 1毛发穿孔试验阳性表明林生地霉和白地霉均能导致毛发感染。 2林生地霉较白地霉对毛发的破坏早且严重。 3林生地霉血液株较皮损株对毛发的破坏早且严重。 4年龄越小,毛发越易受破坏。
[Abstract]:Objective: Geotrichum is a yeast like fungus, belonging to the class of semi - rhizomatous fungi, ceraspora, cyclosporin, and Rhizopus, which represents the bacterial strain of Geotrichum candidum. It is a conditional pathogen that causes bronchial infection and can cause systemic disseminated infection. Geotircum sivicola (Geotircum sivicola) is a new species found in the genus Rhizopus, 2002 It was first isolated from the Brazil fruit fly and the India tussah larvae for the first time. In the same year, we first isolated a strain of.2005 year from the skin lesions of children with tinea purulent. Our family separated a strain from the blood of a patient with toxic epidermal necrosis. Li Xiu Li and so on from morphology, nutrition physiology, and molecular biology. From various angles, we preliminarily studied the Geotrichum candidum, and discussed its pathogenicity through animal experiments and hair invasion tests.
In this study, a modified hair perforation experiment was carried out on hair of different age groups. On the one hand, the morphology of the mycelium and spores of the fungi and the spores of the fungi and the fungi of the fungi and the fungi of the fungi of the forest of the fungi and the fungi were observed under the light and electron microscope. The difference of hair infection time between different age groups was evaluated to evaluate the invasive ability of Trichoderma to hair.
Methods: the fungi were identified by the Institute of Microbiology of the Chinese Academy of Microbiology, which were preserved in our family. White earth mold was purchased from the Institute of Microbiology in the Academy of Sciences. The strains of ground molds, the blood strain of the fungi and the suspension of M. Candida were prepared, and the number of spores in the suspension was controlled in the range of 108/ ml to 109/ ml with the blood cell count board. The hair of 4 age groups (less than 2 years, 2~13, 13~19, and 19 years old) were placed in the glucosamine Peptone Agar Medium and then in every year. The hair on the hair of the age group was inoculated with the strain of the forest ground mould, 3 drops of the blood strain of the fungus and 3 drops of the mould suspension in the 370 temperature box, and the hair of each age group was taken as a blank control. The hair of the hair was observed under the light mirror of 10% KOH and the phenol cotton blue staining under the microscope after the inoculation. The morphology of the conidia and the hair saboteurs were further used by the Hitachi S23500N scanning electron microscope to observe the morphological changes of the strain of the fungi, the morphological differences of the blood strains and the milderus mildera, and the structural changes of the infected hair.
Results: the difference in the time of hair infection in different age groups was compared between the blood strains of lyngillus sp., and the adhesion of hairs on hair and the damage of the hair on the hair as the standard of perforation. The difference was significant (P0.05). In addition, the time needed to infect hair from the skin strain, the blood strain or the infection of the hairs was also prolonged with the age, and there was a significant difference (P0.05). The difference between the strains of different strains was 22, and the blood strain, the skin strain and the mildew of the fungi were found in the age groups. 22 the difference was significant (P0.05), in which the time range of hair perforation was 53-64 days, the average was 58.07 + 3.15 days, the time range of hair perforation was 57-66 days, the average was 61.05 + 2.55 days, and the time range of hair perforation was 61-74 days, and the average was 67.11 + 3.78 days. The time of hair infection is the shortest in the blood strain of mildew and the longest time of hair infection is caused by mildew. Therefore, the damage of the blood strain to the hair is stronger than that of the other two strains.
The difference between the different age groups was compared 22. It was found that there was no significant difference between the group and the 2~13 year old group (P0.05), and the 22 differences in the other age groups were significant. The younger the age was, the more easily the hair was infected.
A brush like change can be observed on the edge of hair, whether it is ground mold or white earth mildew. When the damage is serious, the hair surface can be shaped and the edge of hair is uneven. The light mirrors can see more rectangles, oval joint spores and round small spore, abundant sprout spores, a small amount of thick conidia and mycelium. Under the electron microscope, it was observed under electron microscope that the degree of hair damage to hair was more serious than that of mildew, and the strain of the blood of the fungi was more serious than that of the linseed. The mycelium of M. candidum attached only to the surface of the hair and intruded into the crevice of the small fur. The mycelium of the strain of the strain of forest ground moldy is seriously eroded in the cortex, causing a large amount of destruction and shedding of the small fur, causing the flake defect of the hair and the finer hair. The mycelium of the blood strain of the fungi of the forest earth mould can also cause the cleavage or fracture of hair, and the serious damage of the hair medulla.
Conclusion:
1 hair test showed that both Trichoderma viride and Geotrichum candidum could lead to hair infection.
2. The damage of Trichoderma viride to the hair is earlier and more serious.
3. The damage of the blood plant of the Geotrichum candidum was earlier and more serious than that of the skin lesion.
The smaller the 4 age, the more vulnerable the hair is.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R756

【参考文献】