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欧洲乌鸫(Turdus merula)种群血孢子虫流行率和虫血症的灵敏检测方法:PCR-RFLP和定量PCR的价值

Sensitive measure of prevalence and parasitaemia of haemosporidia from European blackbird (Turdus merula) populations: value of PCR-RFLP and quantitative PCR.

作者信息

Bentz S, Rigaud T, Barroca M, Martin-Laurent F, Bru D, Moreau J, Faivre B

机构信息

Equipe Ecologie Evolutive, UMR CNRS 5561 Biogéosciences, Université de Bourgogne, 6 Bd Gabriel, 21000 Dijon, France.

出版信息

Parasitology. 2006 Dec;133(Pt 6):685-92. doi: 10.1017/S0031182006001090. Epub 2006 Sep 4.

Abstract

Haemosporidian parasites are common in birds in which they act as an important selective pressure. While most studies so far have focused on the effect of their prevalence on host life-history traits, no study has measured the effect of parasitaemia. We developed molecular methods to detect, identify and quantify haemosporidia in 2 natural populations of the Blackbird Turdus merula. Three different parasite genotypes were found - 1 Haemoproteus and 2 Plasmodium. A PCR-RFLP screening revealed that only approximately 3% of blackbirds were free of parasites, compared to the 34% of uninfected birds estimated by blood smear screening. A quantitative PCR (q-PCR) assay revealed a weaker parasitaemia in microscopically undetected parasites compared to microscopically detected ones. Large parasitaemia differences were found between parasite species, suggesting either differing parasite life-histories or host resistance. Parasitaemias were also weaker in male hosts, and in urban habitats, suggesting that both host factors (e.g. immunity) and habitat characteristics (e.g. vector availability) may modulate parasite density. Interestingly, these differences in parasitaemia were comparable to differences in parasite prevalence estimated by smear screening. This suggests that previous results obtained by smear screening should be reinterpreted in terms of parasitaemia instead of parasite prevalence.

摘要

血孢子虫寄生虫在鸟类中很常见,它们是一种重要的选择压力。虽然迄今为止大多数研究都集中在其流行率对宿主生活史特征的影响上,但尚无研究测量过虫血症的影响。我们开发了分子方法来检测、鉴定和量化乌鸫(Turdus merula)两个自然种群中的血孢子虫。发现了三种不同的寄生虫基因型——一种血变原虫和两种疟原虫。一项PCR-RFLP筛查显示,只有约3%的乌鸫没有寄生虫,而通过血涂片筛查估计未感染鸟类的比例为34%。定量PCR(q-PCR)分析显示,与显微镜检测到的寄生虫相比,显微镜下未检测到的寄生虫的虫血症较弱。在寄生虫种类之间发现了较大的虫血症差异,这表明要么是寄生虫生活史不同,要么是宿主抵抗力不同。雄性宿主和城市栖息地中的虫血症也较弱,这表明宿主因素(如免疫力)和栖息地特征(如媒介可用性)都可能调节寄生虫密度。有趣的是,这些虫血症差异与涂片筛查估计的寄生虫流行率差异相当。这表明,以前通过涂片筛查获得的结果应以虫血症而非寄生虫流行率来重新解释。

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