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Effects of diethylcarbamazine and ivermectin treatment on Brugia malayi gene expression in infected gerbils (Meriones unguiculatus). 二乙基卡马嗪和伊维菌素对感染沙鼠马来布鲁氏菌基因表达的影响
Pub Date : 2019-01-01 Epub Date: 2019-03-08 DOI: 10.1017/pao.2019.1
Mary J Maclean, W Walter Lorenz, Michael T Dzimianski, Christopher Anna, Andrew R Moorhead, Barbara J Reaves, Adrian J Wolstenholme

Lymphatic filariasis (LF) threatens nearly 20% of the world's population and has handicapped one-third of the 120 million people currently infected. Current control and elimination programs for LF rely on mass drug administration of albendazole plus diethylcarbamazine (DEC) or ivermectin. Only the mechanism of action of albendazole is well understood. To gain a better insight into antifilarial drug action in vivo, we treated gerbils harbouring patent Brugia malayi infections with 6 mg kg-1 DEC, 0.15 mg kg-1 ivermectin or 1 mg kg-1 albendazole. Treatments had no effect on the numbers of worms present in the peritoneal cavity of treated animals, so effects on gene expression were a direct result of the drug and not complicated by dying parasites. Adults and microfilariae were collected 1 and 7 days post-treatment and RNA isolated for transcriptomic analysis. The experiment was repeated three times. Ivermectin treatment produced the most differentially expressed genes (DEGs), 113. DEC treatment yielded 61 DEGs. Albendazole treatment resulted in little change in gene expression, with only 6 genes affected. In total, nearly 200 DEGs were identified with little overlap between treatment groups, suggesting that these drugs may interfere in different ways with processes important for parasite survival, development, and reproduction.

淋巴丝虫病(LF)威胁着世界近20%的人口,目前感染的1.2亿人中有三分之一残疾。目前的LF控制和消除方案依赖于阿苯达唑加乙基卡马嗪(DEC)或伊维菌素的大量给药。只有阿苯达唑的作用机制是很清楚的。为了更好地了解抗丝虫药物在体内的作用,我们用6 mg kg-1 DEC、0.15 mg kg-1伊维菌素或1 mg kg-1阿苯达唑治疗患有专利马来布鲁氏菌感染的沙鼠。治疗对治疗动物腹膜腔内存在的蠕虫数量没有影响,因此对基因表达的影响是药物的直接结果,而不会因寄生虫死亡而复杂化。处理后1天和7天分别收集成虫和微丝虫,分离RNA进行转录组学分析。这个实验重复了三次。伊维菌素治疗产生了最多的差异表达基因(DEGs), 113。DEC处理的温度为61℃。阿苯达唑治疗后基因表达变化不大,仅有6个基因受影响。总共鉴定出近200个deg,治疗组之间几乎没有重叠,这表明这些药物可能以不同的方式干扰对寄生虫生存、发育和繁殖重要的过程。
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引用次数: 7
Molecular evolution in immune genes across the avian tree of life. 鸟类生命之树免疫基因的分子进化。
Pub Date : 2019-01-01 DOI: 10.1017/PAO.2019.3
D. Outlaw, V. W. Walstrom, Haley N. Bodden, Chuan-yu Hsu, Mark A. Arick, D. Peterson
All organisms encounter pathogens, and birds are especially susceptible to infection by malaria parasites and other haemosporidians. It is important to understand how immune genes, primarily innate immune genes which are the first line of host defense, have evolved across birds, a highly diverse group of tetrapods. Here, we find that innate immune genes are highly conserved across the avian tree of life and that although most show evidence of positive or diversifying selection within specific lineages or clades, the number of sites is often proportionally low in this broader context of putative constraint. Rather, the evidence shows a much higher level of negative or purifying selection in these innate immune genes - rather than adaptive immune genes - which is consistent with birds' long coevolutionary history with pathogens and the need to maintain a rapid response to infection. We further explored avian responses to haemosporidians by comparing differential gene expression in wild birds (1) uninfected with haemosporidians, (2) infected with Plasmodium, and (3) infected with Haemoproteus (Parahaemoproteus). We found patterns of significant differential expression with some genes unique to infection with each genus and a few shared between "treatment" groups, but none that overlapped with the genes included in the phylogenetic study.
所有生物体都会遇到病原体,鸟类特别容易受到疟疾寄生虫和其他血吸虫的感染。了解免疫基因,主要是作为宿主第一道防线的先天免疫基因,是如何在鸟类这一高度多样化的四足动物群体中进化的,这一点很重要。在这里,我们发现先天免疫基因在整个鸟类生命树中是高度保守的,尽管大多数显示出在特定谱系或分支中积极或多样化选择的证据,但在这种假定约束的更广泛背景下,位点的数量通常是成比例的低。相反,证据表明,在这些先天免疫基因(而不是适应性免疫基因)中存在更高水平的负性或净化性选择,这与鸟类与病原体的长期共同进化历史以及保持对感染的快速反应的需要是一致的。通过比较野生鸟类(1)未感染血孢子虫、(2)感染疟原虫和(3)感染嗜血杆菌(副嗜血杆菌)的差异基因表达,我们进一步探讨了鸟类对血孢子虫的反应。我们发现每个属感染特有的一些基因的显著差异表达模式和“治疗”组之间共享的一些基因,但没有与系统发育研究中包含的基因重叠。
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引用次数: 2
A multiplex PCR test to identify four common cattle-adapted Cryptosporidium species - CORRIGENDUM 鉴定四种常见的牛适应隐孢子虫的多重PCR试验-勘误
Pub Date : 2019-01-01 DOI: 10.1017/PAO.2018.16
S. Thomson, E. Innes, N. Jonsson, F. Katzer
© Cambridge University Press 2019. This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited. A multiplex PCR test to identify four common cattle-adapted Cryptosporidium species CORRIGENDUM
©剑桥大学出版社2019。这是一篇开放获取的文章,在知识共享署名许可(http://creativecommons.org/licenses/ by/4.0/)的条款下分发,该许可允许在任何媒介上不受限制地重复使用、分发和复制,前提是正确引用原始作品。鉴定四种常见的牛适应隐孢子虫的多重PCR试验
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引用次数: 3
ROS regulate differentiation of visceralizing Leishmania species into the virulent amastigote form. 活性氧调节内脏利什曼原虫分化为毒力强的无尾虫。
Pub Date : 2018-01-01 Epub Date: 2018-11-06 DOI: 10.1017/pao.2018.15
Yousuf A Khan, Norma W Andrews, Bidyottam Mittra

Leishmania virulence and disease development critically depends on the ability of Leishmania promastigotes to infect, differentiate into amastigote forms and replicate inside mammalian host macrophages. Understanding changes associated with amastigote differentiation in axenic culture conditions is key to identifying virulence factors. Here we compared efficiency of the conventional pH-temperature-dependent shift method to induce amastigote differentiation with the recently identified trigger for differentiation mediated by mitochondrial reactive oxygen species (ROS). Using two different visceral leishmaniasis species, L. infantum and. L. donovani, we show that ROS-generating methods such as iron deprivation or exposure to sub-lethal concentrations of H2O2 or menadione are significantly more effective in promoting promastigoteamastigote differentiation than the low pH-high temperature shift, leading to higher survival rates, morphological changes and gene expression patterns characteristic of the amastigote stage. Notably, both H2O2 and menadione-mediated differentiation did not require up-regulation of the mitochondrial electron transport chain (ETC)-associated protein p27, suggesting that treatment with oxidants bypasses the necessity to upregulate mitochondrial activity, a precondition for mROS generation. Our findings confirm that ROS-induced differentiation occurs in multiple Leishmania species, including the medically important visceralizing species, and provide mechanistic rationale for earlier reports demonstrating markedly increased virulence of L. infantum promastigotes pre-treated with oxidative reagents.

利什曼原虫的毒力和疾病发展在很大程度上取决于利什曼原虫原鞭毛体感染、分化为无鞭毛体形式和在哺乳动物宿主巨噬细胞内复制的能力。了解与无性系培养条件下无性系分化相关的变化是确定毒力因素的关键。在这里,我们比较了传统的ph -温度依赖转移方法诱导无纺体分化的效率,以及最近发现的由线粒体活性氧(ROS)介导的分化触发器。使用两种不同的内脏利什曼病,婴儿乳杆菌和。L. donovani,我们发现产生ros的方法,如铁剥夺或暴露于亚致死浓度的H2O2或甲氧二酮,比低ph -高温转换更有效地促进promastigoteamastigote分化,导致更高的存活率,形态变化和基因表达模式的特征。值得注意的是,H2O2和甲氧二酮介导的分化都不需要上调线粒体电子传递链(ETC)相关蛋白p27,这表明用氧化剂治疗绕过了上调线粒体活性的必要性,而上调线粒体活性是产生mROS的先决条件。我们的研究结果证实,ros诱导的分化发生在多个利什曼原虫物种中,包括医学上重要的内脏化物种,并为早期报告提供了机制基础,证明经氧化试剂预处理的婴儿利什曼原虫的毒力显着增加。
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引用次数: 13
The mitochondrial genomes of the mesozoans Intoshia linei, Dicyema sp. and Dicyema japonicum. 中低等动物的线粒体基因组研究。
Pub Date : 2018-01-01 Epub Date: 2018-08-02 DOI: 10.1017/pao.2018.12
Helen E Robertson, Philipp H Schiffer, Maximilian J Telford

The Dicyemida and Orthonectida are two groups of tiny, simple, vermiform parasites that have historically been united in a group named the Mesozoa. Both Dicyemida and Orthonectida have just two cell layers and appear to lack any defined tissues. They were initially thought to be evolutionary intermediates between protozoans and metazoans but more recent analyses indicate that they are protostomian metazoans that have undergone secondary simplification from a complex ancestor. Here we describe the first almost complete mitochondrial genome sequence from an orthonectid, Intoshia linei, and describe nine and eight mitochondrial protein-coding genes from Dicyema sp. and Dicyema japonicum, respectively. The 14,247 base pair long I. linei sequence has typical metazoan gene content, but is exceptionally AT-rich, and has a unique gene order. The data we have analysed from the Dicyemida provide very limited support for the suggestion that dicyemid mitochondrial genes are found on discrete mini-circles, as opposed to the large circular mitochondrial genomes that are typical of the Metazoa. The cox1 gene from dicyemid species has a series of conserved, in-frame deletions that is unique to this lineage. Using cox1 genes from across the genus Dicyema, we report the first internal phylogeny of this group.

双尾虫和直孔虫是两组微小的、简单的蠕虫状寄生虫,它们在历史上被统一在一个名为Mesozoa的群体中。双叶虫和正孔虫都只有两层细胞,似乎没有任何明确的组织。它们最初被认为是介于原生动物和后生动物之间的进化中间体,但最近的分析表明,它们是由一个复杂的祖先经历了二次简化的原生生动物后生动物。在这里,我们描述了第一个几乎完整的线粒体基因组序列,来自正连科植物Intoshia linei,并分别描述了来自Dicyema sp.和Dicyema japonicum的9个和8个线粒体蛋白质编码基因。全长14247个碱基对的I. linei序列具有典型的后生动物基因含量,但具有异常丰富的at,并具有独特的基因序列。我们从双尾虫身上分析的数据非常有限地支持了双尾虫线粒体基因在离散的小圆圈上发现的建议,而不是典型的后生动物的大圆形线粒体基因组。来自双叶属物种的cox1基因具有一系列保守的框架内缺失,这是该谱系所独有的。利用来自整个双叶菊属的cox1基因,我们报道了这个群体的第一个内部系统发育。
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引用次数: 2
Trypanosoma amblyommi sp. nov. (Protozoa: Kinetoplastida) isolated from Amblyomma brasiliense (Acari: Ixodidae) ticks in Rio de Janeiro, Brazil. 从巴西里约热内卢的Amblyomma brasiliense(Acari: Ixodidae)蜱中分离出的Trypanosoma amblyommi sp.
Pub Date : 2018-01-01 Epub Date: 2018-01-11 DOI: 10.1017/pao.2017.17
Carolina R Marotta, Priscilla N Dos Santos, Matheus D Cordeiro, Juliana Helena Da S Barros, Lesley Bell-Sakyi, Adivaldo H Fonseca

Parasites of the genus Trypanosoma are microorganisms that display wide morphological, biological and genetic variability. Here we present the first description of an isolate of the genus Trypanosoma naturally infecting the tick Amblyomma brasiliense. The ticks were collected from a specimen of Tayassu pecari (Queixada, white-lipped peccary) from the Itatiaia National Park, Itatiaia, Rio de Janeiro, Brazil. The isolate was characterised by molecular, morphometric and biological analyses. A Trypanosoma culture was isolated from crushed nymphal and adult ticks, propagated in the tick cell line IDE8 and maintained in L15B culture medium, incubated at 32 °C. The isolate grew well in L15B medium at 30 °C, 32 °C and 34 °C but not at lower or higher temperatures. The culture remained stable in axenic L15B medium at 30 °C. Cryopreserved cultures retained viability after cryopreservation in liquid nitrogen. Growth in axenic medium and developmental forms of the trypanosomes were analysed. Analysis of the 18S rDNA region confirmed the authenticity of this new species and the nucleotide sequence was deposited in Genbank. The species was named Trypanosoma amblyommi sp. nov. strain C1RJ. Characteristics related to pathogenicity, involvement with vertebrate hosts, epidemiology, developmental cycle and transmission mechanisms are still unknown. Therefore, further studies are necessary to understand aspects of the biological cycle of Trypanosoma amblyommi sp. nov.

锥虫属寄生虫是一种在形态、生物学和遗传学方面具有广泛变异性的微生物。在这里,我们首次描述了自然感染蜱(Amblyomma brasiliense)的锥虫属分离株。蜱虫是从巴西里约热内卢伊塔提亚国家公园的 Tayassu pecari(Queixada,白唇山雀)标本中采集的。通过分子、形态和生物学分析确定了分离物的特征。从压碎的若虫和成蜱中分离出锥虫培养物,在蜱细胞系 IDE8 中繁殖,并在 L15B 培养基中培养,在 32 °C 下孵育。该分离株在 30 ℃、32 ℃ 和 34 ℃ 的 L15B 培养基中生长良好,但在较低或较高温度下生长不良。培养物在 30 ℃ 的腋生 L15B 培养基中保持稳定。低温保存的培养物在液氮中低温保存后仍保持活力。对锥虫在轴突培养基中的生长和发育形式进行了分析。对 18S rDNA 区域的分析证实了这一新物种的真实性,并将核苷酸序列存入 Genbank。该物种被命名为 Trypanosoma amblyommi sp.目前还不清楚该种的致病性、与脊椎动物宿主的关系、流行病学、发育周期和传播机制等方面的特征。因此,有必要开展进一步研究,以了解新伏锥虫的生物周期。
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