Free-living amoebae (FLA) include many genera which cause serious diseases such as sight-threatening keratitis, cutaneous ulcers and fatal encephalitis. This study was conducted due to the lack of research regarding genotypes Acanthamoeba, Naegleria and Vermamoeba in mineral springs of Guilan Province in northern Iran. Twenty-five water samples were collected from mineral springs in Guilan Province. After filtration through nitrocellulose membrane, samples were cultured on non-nutrient agar plates. The morphological key of Page was used to identify free-living amoebae (FLA) using an inverted microscope. Positive cultures were analyzed by polymerase chain reaction (PCR) and genotypes based on the NCBI database. Eleven (44%) samples were positive for Acanthamoeba, Naegleria and Vermamoeba. By sequencing the positive isolates, the strains were shown to belong to Acanthamoeba castellanii (three case isolates belonged to T4 genotype), three cases of Vermamoeba vermiformis, and two cases of N. australiensis, two cases of N. pagei and one case of N. gruberi. According to our research the occurrence of Acanthamoeba, Naegleria spp. and Vermamoeba spp. in mineral springs could be hazardous for high risk persons. Regular monitoring and posting warning signs of these waters by health planners could prevent free-living amoebae mediated diseases.

中文翻译：

---

伊朗北部桂兰省矿泉中的自由活动变形虫（Naegleria spp。，Acanthamoeba spp。和Vermamoeba spp。）的分离和分子鉴定。

自由生活的变形虫（FLA）包括许多会导致严重疾病的物种，例如威胁视力的角膜炎，皮肤溃疡和致命性脑炎。进行这项研究是由于缺乏对伊朗北部桂兰省矿泉中棘形棘阿米巴，Naegleria和Vermamoeba基因型的研究。从桂兰省的矿泉中采集了25个水样。通过硝酸纤维素膜过滤后，将样品在非营养琼脂平板上培养。使用倒置显微镜将Page的形态键用于识别自由活动的变形虫（FLA）。通过聚合酶链反应（PCR）和基于NCBI数据库的基因型分析阳性培养物。十一（44％）样品的棘阿米巴，奈格勒菌和Vermamoeba呈阳性。通过对阳性分离物进行测序，结果表明，该菌株属于卡氏棘阿米巴菌（3个菌株属于T4基因型），3个Vermamoeba vermiformis，2例澳大利亚猪笼草，2例Pagei猪笼草和1例N. gruberi猪笼草。根据我们的研究，棘孢菌，内格氏菌属的发生。和Vermamoeba spp。在矿泉水中可能对高危人群有害。卫生计划人员定期监视并张贴这些水的警告信号，可以预防自由生活的变形虫介导的疾病。在矿泉水中可能对高危人群有害。卫生计划人员定期监视并张贴这些水的警告信号，可以预防自由生活的变形虫介导的疾病。在矿泉水中可能对高危人群有害。卫生计划人员定期监视并张贴这些水的警告信号，可以预防自由生活的变形虫介导的疾病。