Several thermal areas, also used for leisure purposes, may represent suitable habitats for free-living amoebae (FLAs), but few studies have been carried out in search for these organisms. The aim of this study was to assess the presence and distribution of FLAs by culture detection and molecular identification, over a one year-round sampling of two sites in Central Italy. Two geothermal springs (Site A and Site B) were investigated for a total of 36 water samples. Four sets of primers were used to amplify FLA DNA from all cultures positive for amoebic growth at both 37 °C and 45 °C. Overall, 33 (91.6%) water samples produced PCR amplification. Eleven taxa were identified. The array of identified species varied over the sampling period, and differed between the two hot springs, Site A harbouring 11 taxa compared to 5 of site B. However, both sites were characterized by the most common species Vermamoeba vermiformis and Naegleria australiensis. Acanthamoeba genotypes T4 and T15 were found at low frequency. Differences in the composition between the two sites could reflect environmental changes in biotic and chemical/physical parameters. From a public health perspective, the detection of potentially pathogenic amoebae could unveil a potential risk for humans.

几个也用于休闲目的的热区可能是自由生活变形虫 (FLA) 的合适栖息地，但很少有研究用于寻找这些生物。本研究的目的是通过培养检测和分子鉴定评估 FLA 的存在和分布，在意大利中部的两个地点进行为期一年的采样。研究了两个地热泉（A 点和 B 点），共采集了 36 个水样。使用四组引物从所有在 37°C 和 45°C 下对阿米巴生长呈阳性的培养物中扩增 FLA DNA。总体而言，33 (91.6%) 个水样产生了 PCR 扩增。确定了 11 个分类群。已识别物种的排列在采样期间各不相同，并且在两个温泉之间有所不同，站点 A 包含 11 个分类群，而站点 B 包含 5 个。然而，Vermamoeba vermiformis和Naegleria australiensis。棘阿米巴基因型 T4 和 T15 发现频率较低。两个地点之间的组成差异可以反映生物和化学/物理参数的环境变化。从公共卫生的角度来看，潜在致病性变形虫的检测可能会揭示对人类的潜在风险。