Small subunit ribosomal metabarcoding reveals extraordinary trypanosomatid diversity in Brazilian bats.
Small subunit ribosomal metabarcoding reveals extraordinary trypanosomatid diversity in Brazilian bats.
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DOI：
10.1371/journal.pntd.0005790
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发表时间：
2017-07
影响因子：
3.8
通讯作者：
Llewellyn MS
中科院分区：
文献类型：
--
作者：
Dario MA;Moratelli R;Schwabl P;Jansen AM;Llewellyn MS
蝙蝠是一种非常成功的哺乳动物，分布在全球各地，占据着广泛的生态位。它们也被强烈寄生，并与多种病毒、细菌和寄生性人畜共患病有关。锥虫被认为在蝙蝠中特别丰富和多样化。在这项研究中，我们使用18S核糖体RNA元编码来前所未有地详细地探测蝙蝠锥虫的多样性。从巴西东南部圣埃斯皮里托州大西洋森林地区捕获的90只蝙蝠(17种)的血液中提取了总DNA。用标准和/或套式聚合酶链式反应扩增18S核糖体RNA，然后进行深度测序以恢复和鉴定用于系统发育分析的可操作分类单位(OTUS)。13种34只蝙蝠的血液样本经18S rRNA扩增检测呈阳性。扩增序列聚为14个OTUS，其中5个被鉴定为克氏锥虫I、克氏锥虫III/V、马林克氏锥虫、兰氏锥虫和迪奥尼西锥虫，7个被鉴定为新大陆克氏锥虫的单系至基本分支类型。另一种OTU被鉴定为锥虫，与爬行动物中发现的锥虫相似。令人惊讶的是，剩下的OTU被确定为Bodo Saltans--与锥虫目最接近的非寄生近亲。虽然三个血样只有一个OTU(杜氏毛滴虫)，但所有其他血样都被诊断为多达八个OTU的混合感染。这项研究展示了下一代条形码方法在哺乳动物宿主寄生虫多样性筛选中的应用。我们揭露了多种锥虫物种对当地蝙蝠的高寄生率，其中一些已知会导致致命的人类疾病，另一些是非致病的、新颖的或鲜为人知的。我们的结果突出表明，蝙蝠是多个生态位的宿主和寄生虫相互作用的长期纽带，部分原因是对进化论和公共卫生造成后果的机会性和偶发感染。蝙蝠构成了一种极其多样化、高度寄生的游动哺乳动物，它们独特的行为和生理适应能力促进了大量微生物的感染。锥虫是蝙蝠中古老的原生动物寄生虫。针对巴西东南部大西洋森林地区蝙蝠形态隐蔽、寄生虫血症低、选择性生长等问题，采用18S核糖体RNAMetabararCoding技术对蝙蝠的多样性进行了解析。除了几个未知物种外，我们的基于深度序列的检测和分配方案识别了多个已知的人类致病锥虫，另一个与爬行动物宿主以及各种叶口蝙蝠血液中的非寄生动体相关联。在这一地区，蝙蝠锥虫最近出现了一例查加斯病的致命病例，这里暴露出的惊人的容许性，迫使人们进一步研究蝙蝠在各种微生物在人类和野生动物中的传播和溢出中所扮演的角色。
Bats are a highly successful, globally dispersed order of mammals that occupy a wide array of ecological niches. They are also intensely parasitized and implicated in multiple viral, bacterial and parasitic zoonoses. Trypanosomes are thought to be especially abundant and diverse in bats. In this study, we used 18S ribosomal RNA metabarcoding to probe bat trypanosome diversity in unprecedented detail. Total DNA was extracted from the blood of 90 bat individuals (17 species) captured along Atlantic Forest fragments of Espírito Santo state, southeast Brazil. 18S ribosomal RNA was amplified by standard and/or nested PCR, then deep sequenced to recover and identify Operational Taxonomic Units (OTUs) for phylogenetic analysis. Blood samples from 34 bat individuals (13 species) tested positive for infection by 18S rRNA amplification. Amplicon sequences clustered to 14 OTUs, of which five were identified as Trypanosoma cruzi I, T. cruzi III/V, Trypanosoma cruzi marinkellei, Trypanosoma rangeli, and Trypanosoma dionisii, and seven were identified as novel genotypes monophyletic to basal T. cruzi clade types of the New World. Another OTU was identified as a trypanosome like those found in reptiles. Surprisingly, the remaining OTU was identified as Bodo saltans–closest non-parasitic relative of the trypanosomatid order. While three blood samples featured just one OTU (T. dionisii), all others resolved as mixed infections of up to eight OTUs. This study demonstrates the utility of next-generation barcoding methods to screen parasite diversity in mammalian reservoir hosts. We exposed high rates of local bat parasitism by multiple trypanosome species, some known to cause fatal human disease, others non-pathogenic, novel or yet little understood. Our results highlight bats as a long-standing nexus among host-parasite interactions of multiple niches, sustained in part by opportunistic and incidental infections of consequence to evolutionary theory as much as to public health. Bats make up a mega-diverse, intensely parasitized order of volant mammals whose unique behavioural and physiological adaptations promote infection by a vast array of microorganisms. Trypanosomes stand out as ancient protozoan parasites of bats. As cryptic morphology, low parasitaemia and selective growth in culture have recurrently biased survey, we used 18S ribosomal RNA metabarcoding to resolve bat trypanosomatid diversity in Atlantic Forest fragments of southeast Brazil. Next to several unknown species, our deep sequence-based detection and assignment protocol recognized multiple known human-pathogenic trypanosomes, another linked to reptile hosts as well as a non-parasitic kinetoplastid in the blood of various phyllostomid bats. The striking permissivity exposed here, in a region where bat trypanosomes recently featured in a fatal case of Chagas disease, compels further research on bats’ role in the dispersal and spill-over of various microorganisms among humans and wildlife.
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影响因子：
15.9
作者：
Brook CE;Dobson AP
通讯作者：
Dobson AP
DOI：
10.1590/s0074-02762006000500010
发表时间：
2006-08-01
期刊：
Memórias do Instituto Oswaldo Cruz
影响因子：
--
作者：
Santos, Claudiney Biral dos;Leite, Gustavo Rocha;Falqueto, Aloísio
通讯作者：
Falqueto, Aloísio
影响因子：
4
作者：
Cavazzana, Manzelio, Jr.;Marcili, Arlei;Teixeira, Marta M. G.
通讯作者：
Teixeira, Marta M. G.
影响因子：
3.7
作者：
Cottontail, Veronika M.;Kalko, Elisabeth K. V.;Miguel Pinto, C.
通讯作者：
Miguel Pinto, C.
影响因子：
2.6
作者：
Barbosa AD;Mackie JT;Stenner R;Gillett A;Irwin P;Ryan U
通讯作者：
Ryan U