加蒂隐球菌VGI/VGII的体外生长特性及其对mellonella的毒力多样性

Q4 Immunology and Microbiology

中华微生物学和免疫学杂志 Pub Date : 2019-10-31 DOI:10.3760/CMA.J.ISSN.0254-5101.2019.10.001

Chen Yang, Liangli Jin, Lifeng Wang, Xinying Xue, D. Shen

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The growth curves of Cryptococcus gattii VGⅠ and VGⅡ were similar to that of Cryptococcus neoformans when culture at 30℃. The total number for each of them could reach 108 CFU/ml at 96 h under 30℃. However, the total number at any time point at 37℃ was less than that at 30℃. There was no significant difference in the amount of melanin produced by Cryptococcus neoformans under 30℃ and 37℃, but both VGⅠand VGⅡ types of Cryptococcus gattii could produce more amount of melanin under 37℃ than under 30℃. The ratio of capsule/cell wall diameter of Cryptococcus gattii VGⅠwas greater at 37℃ than that at 30℃ with statistical significance (P<0.001). Cryptococcus neoformans showed the longest LT50, followed by VGⅠand VGⅡ types of Cryptococcus gattii. 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引用次数: 0

摘要

目的了解我国分离的gattii隐球菌VGⅠ和VGⅡ的体外生长特性及其对梅勒菌毒力的多样性。方法根据我国分离的8株gattii隐球菌的多点序列分型结果，对其进行体外培养，绘制生长曲线，观察黑色素的产生，并测定包膜厚度。用意大利黑加仑菌感染试验计算gattii隐球菌48小时的中位致死时间（LT50）和中位致死剂量（LC50）。对14株新型隐球菌进行了比较研究。结果8株gattii隐球菌分别为6株VGⅠ和2株VGⅡ。gattii隐球菌VGⅠ和VGⅡ在30℃培养时的生长曲线与新生隐球菌相似。在30℃下96h，每株菌落总数可达108CFU／ml。然而，在37℃的任何时间点的总数都小于30℃的总数。新生隐球菌在30℃和37℃下产生的黑色素量没有显著差异，但VGⅠ型和VGⅡ型的gattii隐球菌在37℃下都能产生比30℃下更多的黑色素。gattii隐球菌的包膜/细胞壁直径之比VGⅠ在37℃时大于30℃时，具有统计学意义（P＜0.001）。新生隐球菌表现出最长的LT50，其次是VGⅠ型和VGⅡ型。gattii隐球菌VGⅡ在1×106CFU／ml浓度下的LT50为72 h，48 h的LC50为1×108CFU／ml。结论与新生隐球菌一样，gattii隐球菌VGⅠ和VGⅡ在30℃下比37℃下生长更快，但在37℃下比30℃下产生更多的黑色素，形成更厚的包膜。在新生隐球菌和gattii隐球菌的VGⅠ和VGⅡ型中，gattii隐球菌VGⅡ的LT50最短，对意大利加仑菌的毒力最强。关键词：gattii隐球菌；分子类型；生长特征；意大利广场；毒力

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In vitro growth characteristics of Cryptococcus gattii VGI/VGII and their virulence diversity to Galleria mellonella

Objective To understand the in vitro growth characteristics of Cryptococcus gattii VGⅠ and VGⅡ isolated in China and the diversity in their virulence to Galleria mellonella. Methods Based on the results of multilocus sequence typing for eight strains of Cryptococcus gattii isolated in China, the strains were cultured in vitro to draw growth curves, observe the melanin production and measure the capsule thickness. The median lethal time (LT50) and median lethal dose (LC50) at 48 h of Cryptococcus gattii were calculated using Galleria mellonella infection test. Fourteen strains of Cryptococcus neoformans were studied for comparison. Results The eight Cryptococcus gattii strains were six VGⅠ and two VGⅡ. The growth curves of Cryptococcus gattii VGⅠ and VGⅡ were similar to that of Cryptococcus neoformans when culture at 30℃. The total number for each of them could reach 108 CFU/ml at 96 h under 30℃. However, the total number at any time point at 37℃ was less than that at 30℃. There was no significant difference in the amount of melanin produced by Cryptococcus neoformans under 30℃ and 37℃, but both VGⅠand VGⅡ types of Cryptococcus gattii could produce more amount of melanin under 37℃ than under 30℃. The ratio of capsule/cell wall diameter of Cryptococcus gattii VGⅠwas greater at 37℃ than that at 30℃ with statistical significance (P<0.001). Cryptococcus neoformans showed the longest LT50, followed by VGⅠand VGⅡ types of Cryptococcus gattii. The LT50 of Cryptococcus gattii VGⅡ at the concentration of 1×106 CFU/ml was 72 h, and its LC50 at 48 h was 1×108 CFU/ml. Conclusions Like Cryptococcus neoformans, Cryptococcus gattii VGⅠ and VGⅡ grew faster under 30℃ than under 37℃, but more melanin was produced and thicker capsule was formed under 37℃ than under 30℃. Among Cryptococcus neoformans and VGⅠ and VGⅡ types of Cryptococcus gattii, Cryptococcus gattii VGⅡ showed the shortest LT50 and the strongest virulence to Galleria mellonella. Key words: Cryptococcus gattii; Molecular type; Growth characteristics; Galleria mellonella; Virulence

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来源期刊

中华微生物学和免疫学杂志 Immunology and Microbiology-Virology

CiteScore

0.50

自引率

0.00%

发文量

6906

期刊介绍： Chinese Journal of Microbiology and Immunology established in 1981. It is one of the series of journal sponsored by Chinese Medical Association. The aim of this journal is to spread and exchange the scientific achievements and practical experience in order to promote the development of medical microbiology and immunology. Its main contents comprise academic thesis, brief reports, reviews, summaries, news of meetings, book reviews and trends of home and abroad in this field. The distinguishing feature of the journal is to give the priority to the reports on the research of basic theory, and take account of the reports on clinical and practical skills.