Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran.
Department of Microbiology, Tonekabon Branch, Islamic Azad University, Tonekabon, Iran.
Pol J Microbiol. 2020 Sep;69(3):301-310. doi: 10.33073/pjm-2020-033. Epub 2020 Sep 8.
Leptospirosis is a worldwide infectious and zoonotic disease. The incidence of this disease is high in temperate regions, especially in northern Iran. The aim of this study was to investigate the effects of temperature, pH, and plant extract on the gene expression in pathogenic spp. Fifty water samples were collected. Culture and PCR technique were used to isolate and identify the bacterium and the presence of the gene. The samples were exposed to different temperatures and pH levels for one day and the plant extract at different concentrations for one and seven days. RNA was extracted, and cDNA synthesis was performed for all the samples. All cDNAs were evaluated by the real-time PCR (SYBR green) technique. Out of the 50 samples, ten samples (20%), using PCR were determined to contain the pathogenic . Fold change of the expression of the gene associated with stresses was as follows: temperature stress of 40°C, 35°C, and 25°C reduced the gene expression in all three isolates, especially in the isolates type 1. The pH stress, i.e., pH values equal to 8 or 9 reduced the gene expression in three types of isolates, and pH = 6 stress increases the gene expression in the isolates of type 1. plant extract stress reduced the mentioned gene expression only in isolates of type 2. Temperature and pH stresses could lead to differences in the expression level and cause the gene expression decrease in three pathogenic isolates. The MIC results showed anti-leptospiral effect of plant extract.
钩端螺旋体病是一种全球性的传染性人畜共患病。在温带地区,尤其是在伊朗北部,该病的发病率较高。本研究旨在探讨温度、pH 值和植物提取物对致病 spp.基因表达的影响。采集了 50 个水样。采用培养和 PCR 技术分离和鉴定细菌,并检测 基因的存在。将样品暴露于不同的温度和 pH 值下一天,以及不同浓度的 植物提取物下一天和七天。提取 RNA,对所有样品进行 cDNA 合成。所有 cDNA 均采用实时 PCR(SYBR Green)技术进行评估。在 50 个样本中,使用 PCR 检测到 10 个样本(20%)含有致病性 。与应激相关的 基因表达的倍数变化如下:40°C、35°C 和 25°C 的温度应激降低了所有三种分离株中 基因的表达,尤其是在 1 型分离株中。pH 值应激,即 pH 值等于 8 或 9,降低了三种分离株中基因的表达,而 pH = 6 的应激增加了 1 型分离株中 基因的表达。植物提取物应激仅降低了 2 型分离株中所述基因的表达。温度和 pH 值应激可导致表达水平的差异,并导致三种致病性分离株中 基因表达减少。MIC 结果显示植物提取物具有抗钩端螺旋体作用。