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Quantitative Estimation of the Viability of Toxoplasma gondii Oocysts in Soil

机译:弓形虫卵囊在土壤中生存力的定量估计

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Toxoplasma gondii oocysts spread in the environment are an important source of toxoplasmosis for humans and animal species. Although the life expectancy of oocysts has been studied through the infectivity of inoculated soil samples, the survival dynamics of oocysts in the environment are poorly documented. The aim of this study was to quantify oocyst viability in soil over time under two rain conditions. Oocysts were placed in 54 sentinel chambers containing soil and 18 sealed water tubes, all settled in two containers filled with soil. Containers were watered to simulate rain levels of arid and wet climates and kept at stable temperature for 21.5 months. At nine sampling dates during this period, we sampled six chambers and two water tubes. Three methods were used to measure oocyst viability: microscopic counting, quantitative PCR (qPCR), and mouse inoculation. In parallel, oocysts were kept refrigerated during the same period to analyze their detectability over time. Microscopic counting, qPCR, and mouse inoculation all showed decreasing values over time and highly significant differences between the decreases under dry and damp conditions. The proportion of oocysts surviving after 100 days was estimated to be 7.4% (95% confidence interval [95% CI] = 5.1, 10.8) under dry conditions and 43.7% (5% CI = 35.6, 53.5) under damp conditions. The detectability of oocysts by qPCR over time decreased by 0.5 cycle threshold per 100 days. Finally, a strong correlation between qPCR results and the dose infecting 50% of mice was found; thus, qPCR results may be used as an estimate of the infectivity of soil samples.
机译:弓形虫卵囊在环境中传播是人类和动物弓形虫病的重要来源。尽管已经通过接种土壤样品的感染性研究了卵囊的预期寿命,但是关于卵囊在环境中的生存动态的文献却很少。这项研究的目的是量化两种雨天条件下土壤中卵囊生存力。将卵囊放置在装有土壤的54个前哨室和18个密封的水管中,全部放置在两个装有土壤的容器中。给容器浇水以模拟干旱和潮湿气候下的降雨水平,并在稳定温度下保持21.5个月。在此期间的九个采样日期,我们采样了六个隔室和两个水管。三种方法用于测量卵囊生存力:显微镜计数,定量PCR(qPCR)和小鼠接种。同时,将卵囊在同一时期冷藏,以分析其随时间的可检测性。显微计数,qPCR和小鼠接种均显示出随着时间的推移值降低,并且在干燥和潮湿条件下的降低之间存在极显着的差异。在干燥条件下,存活100天后的卵囊比例估计为7.4%(95%置信区间[95%CI] = 5.1,10.8),在潮湿条件下为43.7%(5%CI = 35.6,53.5)。随着时间的流逝,qPCR对卵囊的可检测性每100天降低了0.5个循环阈值。最终,发现qPCR结果与感染50%小鼠的剂量之间有很强的相关性。因此,qPCR结果可以用作土壤样品传染性的估计。

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