线虫模型中气单胞菌感染的毒力及发病机制评价
Summary
在这里, 我们介绍了三个不同的实验来研究空气单胞菌感染的线虫。利用这些方便的方法, 很容易评估aeromonas物种之间和内部的毒性。
Abstract
人体病原体气单胞菌已被临床证明可引起肠胃炎、伤口感染、败血症和尿路感染。据报道, 大多数人类疾病与四种细菌有关:沙科氏气菌、嗜水气单胞菌、马尖气单胞菌和鱼子酱。菌型菌是一种具有良好感染模型的细菌, 为研究气单胞菌的细菌发病机制提供了良好的感染模型。在这里, 我们介绍了三个不同的实验, 以研究气单胞菌感染使用线虫模型, 包括生存, 液体毒性, 和肌肉坏死检测。三种测定气单胞菌毒力的方法的结果是一致的。在引起临床感染的4种主要aeromonas物种中,达克斯被认为是毒性最大的物种。这些方法被证明是一个方便的方法来评估气单胞菌物种之间和内部的毒性, 并有助于我们了解气单胞菌感染的发病机制。
Introduction
人体病原体气单胞菌已被临床证明会引起肠胃炎、伤口感染、败血症和尿路感染1, 2.据报告, 大多数相关的人类疾病与四种细菌有关: dhakensis 气菌、水氢气单胞菌、veronii型和caviae 气体 2,3,4 个,5.在气单胞菌传染病中, 软组织感染可导致人类严重发病率和死亡率。值得注意的是, 肌肉坏死是软组织感染最严重的形式.观察线虫感染后的生存和肌肉坏死是推测气单胞菌毒性的一种简便方法。
科学家们已经开发出许多模型生物来研究细菌感染。在以往的研究中, 小鼠、斑马鱼和线虫被用作动物模型, 研究aeromonas6、7、8的发病机制和毒力。每一种动物模型都有其优势和应用。该生物模型, 线虫, 是一种具有天然的细菌摄入的细菌线虫。线虫在进化过程中已经形成了一种复杂的抗细菌感染的先天免疫系统。在细菌感染的压力下,线虫已被证明是研究aeromonas6、7、9等病原体细菌发病机制的良好感染模型如真菌10和肠出血性大肠杆菌o157: h7 11.然而, 目前还没有一份出版物关注使用作为研究 aeromonas 毒力的模型的方法.
在这里, 我们介绍了三个不同的实验, 以研究aeromonas感染使用线虫作为动物模型: 检测生存, 液体毒性, 和肌肉坏死. 这些方法是评价气单胞菌物种之间和内部毒性的一种简便方法, 有助于提高对气单胞菌发病机制的认识。
Protocol
Representative Results
Discussion
线虫是一种天然摄入细菌作为食物的细菌的细菌线虫, 在其进化过程中对细菌产生了复杂的先天免疫力。维持和支持免疫的两个主要器官是表皮和肠道 9,13。线虫的表皮和肌肉带类似于哺乳动物和人类的软组织结构.由于这些特点,线虫可作为研究气单胞菌感染发病机制的模型生物。
在这里…
Disclosures
The authors have nothing to disclose.
Acknowledgements
我们感谢台湾线虫核心设施的协助, 感谢国立成功大学医院微生物学和抗菌素耐药性诊断实验室提供的 aeromonas分离物。我们也感谢 caenorhabditis 遗传学中心 (cgc) 和蠕虫基地。我们也感谢萨瓦娜·摩尔编辑了手稿。
这项研究得到了台湾科技部 (最大部分 105-268-b-0017-017-my3) 和国立成功大学医院 (nckuh-10705001) 至 p. l. chen 的部分资助。
Materials
| Shaker incubator | YIH DER | LM-570R | Bacteria incubation |
| K2HPO4 | J.T.Baker | MP021519455 | Culture medium preparation |
| KH2PO4 | J.T.Baker | 3246-05 | Culture medium preparation |
| Na2HPO4 | J.T.Baker | MP021914405 | Culture medium preparation |
| NaCl | SIGMA | 31434 | Culture medium preparation |
| MgSO4 | SIGMA | M7506 | Culture medium preparation |
| agar | Difco | 214530 | Culture medium preparation |
| CaCl2 | SIGMA | C1016 | Culture medium preparation |
| cholesterol | SIGMA | C8503 | Culture medium preparation |
| ethanol | SIGMA | 32205 | Culture medium preparation |
| KOH | SIGMA | P5958 | Culture medium preparation |
| 6 cm petri plate | ALPHA PLUS | 46 | agar plate preparation |
| 96-well plate | FALCON | 353072 | liquid assay |
| bacterial peptone | Affymetrix/USB | AAJ20048P2 | Culture medium preparation |
| yeast extract | SIGMA | 92144 | Culture medium preparation |
| citric acid•H2O | SIGMA | C1909 | Culture medium preparation |
| tri-potassium citrate•H2O | SIGMA | 104956 | Culture medium preparation |
| FudR | SIGMA | 1271008 | Culture medium preparation |
| disodium EDTA | SIGMA | E1644 | Culture medium preparation |
| FeSO4•7 H2O | SIGMA | 215422 | Culture medium preparation |
| MnCl2•4 H2O | SIGMA | 221279 | Culture medium preparation |
| ZnSO4•7 H2O | SIGMA | 204986 | Culture medium preparation |
| CuSO4•5 H2O | SIGMA | C8027 | Culture medium preparation |
| tryptone | SIGMA | 16922 | Culture medium preparation |
| Microscope system | Nikon | Eclipase Ti inverted | microscope imaging |
| Scientific CCD Camera | QImaging | Retiga-2000R Fast 1394 | microscope imaging |
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