Molecular Detection of Toxoplasma gondii Oocytes in Soil Samples from Guilan Province, Northern Iran

Hossien Hassani Lafmejan Pour, M. Tavalla, Hossien Keshavarz Valian, Mehdi Mohebali, Saeedeh Hashemi Hafshejani, Ali Latifi, M. Salimi, Aref Teimouri, S. Shojaee
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Abstract

Background: Soil is an appropriate substrate for the storage and transmission of oocytes of Toxoplasma gondii. Ingestion of soil contaminated with T. gondii oocysts is a major transmission route of human and animal toxoplasmosis. The present study was carried out to investigate soil contamination with T. gondii oocysts in urban and rural areas of Guilan Province, northern Iran. Methods: Overall, 208 soil samples were collected from 16 cities and villages in Guilan Province, northern Iran from Oct 2020 to Nov 2021. Soil samples were investigated using modified sucrose flotation technique. Real-time polymerase chain reaction was used to detect presence of T. gondii DNAs in the samples. Positive samples were further analyzed using nested polymerase chain reaction for GRA6 gene. Moreover, six selected positive samples were used for amplifying and sequencing of the GRA6 gene. Results: Overall, 31 samples were positive for T. gondii with frequency of 14.9% and ranging from 10.9% in rural areas to 16.3% in urban areas. Statistical analysis showed significant differences between the seasons (P=0.003). The phylogenetic analysis illustrated that our six sequences were similar and closely related to Type I strain of T. gondii. Conclusion: Results showed relatively high levels (14.9%) of T. gondii oocytes in soil samples of Guilan Province, northern Iran, which provided essential data for the effective prevention and control of toxoplasmosis in the region.
伊朗北部吉兰省土壤样本中弓形虫卵母细胞的分子检测
背景:土壤是弓形虫卵母细胞储存和传播的合适基质。摄入受刚地弓形虫卵囊污染的土壤是人类和动物弓形虫病的主要传播途径。本研究旨在调查伊朗北部吉兰省城市和农村地区受刚地弓形虫卵囊污染的土壤。研究方法2020 年 10 月至 2021 年 11 月期间,在伊朗北部吉兰省的 16 个城市和村庄共采集了 208 份土壤样本。采用改良的蔗糖浮选技术对土壤样本进行调查。采用实时聚合酶链反应检测样本中是否存在淋球菌 DNA。对阳性样本进一步使用巢式聚合酶链反应分析 GRA6 基因。此外,还选取了六个阳性样本对 GRA6 基因进行扩增和测序。结果总体而言,31 个样本对淋病双球菌呈阳性,阳性率为 14.9%,农村地区为 10.9%,城市地区为 16.3%。统计分析显示,不同季节之间存在明显差异(P=0.003)。系统进化分析表明,我们的六个序列与淋病双球菌的 I 型菌株相似且关系密切。结论结果显示,伊朗北部吉兰省土壤样本中的弓形虫卵母细胞含量相对较高(14.9%),这为该地区有效预防和控制弓形虫病提供了重要数据。
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