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Exploring dietary differences among developmental stages of triatomines infected with Trypanosoma cruzi in different habitats 探索不同栖息地感染了克鲁兹锥虫的三足虫各发育阶段的饮食差异。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-01 DOI: 10.1016/j.ijpara.2024.05.001

Chagas disease affects millions of people in Colombia and worldwide, with its transmission influenced by ecological, environmental, and anthropogenic factors. There is a notable correlation between vector transmission cycles and the habitats of insect vectors of the parasite. However, the scale at which these cycles operate remains uncertain. While individual triatomine ecotopes such as palms provide conditions for isolated transmission cycles, recent studies examining triatomine blood sources in various habitats suggest a more intricate network of transmission cycles, linking wild ecotopes with human dwellings. This study aims to provide further evidence on the complexity of the scale of Trypanosoma cruzi transmission cycles, by exploring the different blood sources among developmental stages of infected triatomines in different habitats. We evaluated infection rates, parasite loads, feeding sources, and the distribution of Rhodnius prolixus insects in Attalea butyracea palms across three distinct habitats in Casanare, Colombia: peridomestics, pastures, and woodlands. Our results show that there is no clear independence in transmission cycles in each environment. Analyses of feeding sources suggest the movement of insects and mammals (primarily bats and didelphids) among habitats. A significant association was found between habitat and instar stages in collected R. prolixus. The N1 stage was correlated with pasture and woodland, while the N4 stage was related to pasture. Additionally, adult insects exhibited higher T. cruzi loads than N1, N2, and N3. We observed higher T. cruzi loads in insects captured in dwelling and pasture habitats, compared with those captured in woodland areas. Effective Chagas disease control strategies must consider the complexity of transmission cycles and the interplay between domestic and sylvatic populations of mammals and vectors.

南美锥虫病影响着哥伦比亚和全世界数百万人,其传播受生态、环境和人为因素的影响。病媒传播周期与寄生虫昆虫病媒的栖息地之间存在明显的相关性。然而,这些周期的运作规模仍不确定。虽然棕榈树等个别三蠹生态群落为孤立的传播周期提供了条件,但最近对不同栖息地的三蠹血源进行的研究表明,传播周期网络更加错综复杂,将野生生态群落与人类居住地联系在一起。本研究旨在通过探究不同栖息地中受感染三蠹发育阶段的不同血液来源,进一步证明克氏锥虫传播周期规模的复杂性。我们评估了哥伦比亚卡萨纳雷三种不同栖息地(围牧场、牧场和林地)的感染率、寄生虫量、食源以及Attalea butyracea棕榈中Rhodnius prolixus昆虫的分布情况。我们的研究结果表明,每种环境中的传播周期都没有明显的独立性。对食物来源的分析表明,昆虫和哺乳动物(主要是蝙蝠和蚜虫)在不同的栖息地之间流动。在采集到的 R. prolixus 中,栖息地与分化阶段之间存在明显的关联。N1 阶段与牧场和林地相关,而 N4 阶段与牧场相关。此外,与 N1、N2 和 N3 相比,成虫表现出更高的 T. cruzi 负荷。我们观察到,与在林地捕获的昆虫相比,在住所和牧场捕获的昆虫体内有更高的南美锥虫病病毒载量。有效的南美锥虫病控制策略必须考虑到传播周期的复杂性,以及哺乳动物和病媒在家养种群和草原种群之间的相互作用。
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引用次数: 0
Ghosts of parasites past influence current non-consumptive effects in Drosophila nigrospiracula 过去寄生虫的幽灵影响黑腹果蝇当前的非消耗性效应
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-01 DOI: 10.1016/j.ijpara.2024.04.008

Parasites can indirectly impact hosts through non-consumptive effects (NCEs) via changes in behaviour, morphology, and/or physiology. These responses can be understood in terms of the ecology of fear (ectoparasites) or the ecology of disgust (endoparasites) framework. We tested the hypothesis that NCEs of parasite exposure (e.g., parasite avoidance and defense) trade off with other important behaviours such as feeding and resting. We predicted that when exposed to parasites (without infection), hosts will increase their defensive behaviors at the expense of feeding. We also posited that history of exposure (without infection), or previous infection would impact the expression of these NCEs. The study system involves a cactophilic fruit fly (Drosophila nigrospiracula) and a naturally occurring parasitic mite (Macrocheles subbadius). First, we assessed how prior mite exposure affected fly behaviour in response to current parasite exposure. Mite presence resulted in increased grooming and movement, but exposure history did not affect these behaviours. However, the interaction between previous and current exposure influenced host feeding and resting behaviours. We found that previously exposed flies increased feeding and decreased resting upon a secondary mite exposure. In a second experiment, we tested the role of infection history on current parasite exposure. Compared with naïve flies, previously infected flies were expected to increase defensive behaviours upon secondary exposure. Flies increased defensive and ambulatory behaviour in the presence of mites, and consequently less time was spent resting but feeding was unaffected. None of the behaviours measured were affected by previous infection status. In general, current parasite exposure resulted in NCEs. Moreover, our results showed that previous exposure (without infection) to parasites may have an even stronger effect upon secondary exposure than infection history. Our study highlights the importance of the ecology of fear and the role that exposure and infection history plays in generating NCEs of parasitism.

寄生虫可通过行为、形态和/或生理变化对宿主产生非消费性影响(NCE),从而间接影响宿主。这些反应可以从恐惧生态学(外寄生虫)或厌恶生态学(内寄生虫)的框架来理解。我们测试了一个假设,即暴露于寄生虫环境中的NCE(如躲避寄生虫和防御寄生虫)会与其他重要行为(如进食和休息)发生交换。我们预测,当暴露于寄生虫(未感染)时,宿主会增加防御行为而牺牲进食。我们还假设,接触寄生虫的历史(未感染)或之前的感染会影响这些 NCEs 的表达。研究系统涉及一种嗜仙人掌果蝇(Drosophila nigrospiracula)和一种自然发生的寄生螨(Macrocheles subbadius)。首先,我们评估了先前的螨虫接触如何影响苍蝇对当前寄生虫接触的行为反应。螨虫的存在增加了苍蝇的梳理和运动,但接触史并不影响这些行为。然而,先前和当前暴露之间的相互作用影响了宿主的摄食和休息行为。我们发现,之前接触过螨虫的苍蝇在第二次接触螨虫时会增加摄食,减少休息。在第二个实验中,我们测试了感染史对当前寄生虫暴露的影响。与天真苍蝇相比,之前感染过寄生虫的苍蝇在二次接触寄生虫时会增加防御行为。在有螨虫存在的情况下,苍蝇会增加防御和活动行为,因此休息时间减少,但进食不受影响。所测量的行为均不受先前感染状况的影响。总的来说,当前的寄生虫暴露会导致无行为能力。此外,我们的研究结果表明,以前接触寄生虫(未感染)对二次接触的影响可能比感染史更大。我们的研究凸显了恐惧生态学的重要性,以及暴露和感染史在产生寄生虫非传染性行为中的作用。
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引用次数: 0
Multiple UDP glycosyltransferases modulate benzimidazole drug sensitivity in the nematode Caenorhabditis elegans in an additive manner 多种 UDP 糖基转移酶以相加的方式调节线虫秀丽隐杆线虫对苯并咪唑类药物的敏感性
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-01 DOI: 10.1016/j.ijpara.2024.05.003

Xenobiotic biotransformation is an important modulator of anthelmintic drug potency and a potential mechanism of anthelmintic resistance. Both the free-living nematode Caenorhabditis elegans and the ruminant parasite Haemonchus contortus biotransform benzimidazole drugs by glucose conjugation, likely catalysed by UDP-glycosyltransferase (UGT) enzymes. To identify C. elegans genes involved in benzimidazole drug detoxification, we first used a comparative phylogenetic analysis of UGTs from humans, C. elegans and H. contortus, combined with available RNAseq datasets to identify which of the 63 C. elegans ugt genes are most likely to be involved in benzimidazole drug biotransformation. RNA interference knockdown of 15 prioritized C. elegans genes identified those that sensitized animals to the benzimidazole derivative albendazole (ABZ). Genetic mutations subsequently revealed that loss of ugt-9 and ugt-11 had the strongest effects. The “ugt-9 cluster” includes these genes, together with six other closely related ugts. A CRISPR-Cas-9 deletion that removed seven of the eight ugt-9 cluster genes had greater ABZ sensitivity than the single largest-effect mutation. Furthermore, a double mutant of ugt-22 (which is not a member of the ugt-9 cluster) with the ugt-9 cluster deletion further increased ABZ sensitivity. This additivity of mutant phenotypes suggest that ugt genes act in parallel, which could have several, not mutually exclusive, explanations. ugt mutations have different effects with different benzimidazole derivatives, suggesting that enzymes with different specificities could together more efficiently detoxify drugs. Expression patterns of ugt-9, ugt-11 and ugt-22 gfp reporters differ and so likely act in different tissues which may, at least in part, explain their additive effects on drug potency. Overexpression of ugt-9 alone was sufficient to confer partial ABZ resistance, indicating increasing total UGT activity protects animals. In summary, our results suggest that the multiple UGT enzymes have overlapping but not completely redundant functions in benzimidazole drug detoxification and may represent “druggable” targets to improve benzimidazole drug potency.

异生物的生物转化是抗蠕虫药药效的重要调节因素,也是产生抗蠕虫药耐药性的潜在机制。自由生活的线虫秀丽隐杆线虫(Caenorhabditis elegans)和反刍动物寄生虫柯氏血吸虫(Haemonchus contortus)都通过葡萄糖共轭作用对苯并咪唑类药物进行生物转化,这种作用可能是由 UDP-糖基转移酶(UGT)催化的。为了确定参与苯并咪唑类药物解毒的秀丽隐杆线虫基因,我们首先对来自人类、秀丽隐杆线虫和H. contortus的UGT进行了系统发育比较分析,并结合现有的RNAseq数据集,确定了63个秀丽隐杆线虫ugt基因中哪些基因最有可能参与苯并咪唑类药物的生物转化。通过 RNA 干扰敲除 15 个优先考虑的秀丽隐杆线虫基因,确定了那些能使动物对苯并咪唑衍生物阿苯达唑(ABZ)过敏的基因。随后的基因突变显示,ugt-9 和 ugt-11 基因的缺失影响最大。ugt-9基因簇 "包括这些基因以及其他六个密切相关的ugts基因。通过 CRISPR-Cas-9 基因缺失,去除了 8 个 ugt-9 簇基因中的 7 个,其对 ABZ 的敏感性要高于单一的最大效应突变。此外,ugt-22(不属于 ugt-9 基因簇)与 ugt-9 基因簇缺失的双突变体进一步提高了 ABZ 的敏感性。ugt突变对不同的苯并咪唑衍生物有不同的影响,这表明具有不同特异性的酶可以共同更有效地解毒。ugt-9、ugt-11 和 ugt-22 gfp 报告基因的表达模式不同,因此可能作用于不同的组织,这至少可以部分解释它们对药物效力的叠加效应。单独过表达 ugt-9 就足以产生部分 ABZ 抗性,这表明保护动物的总 UGT 活性在增加。总之,我们的研究结果表明,多种 UGT 酶在苯并咪唑类药物解毒过程中具有重叠但非完全冗余的功能,可能是提高苯并咪唑类药物药效的 "可药物 "靶点。
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引用次数: 0
GRA47 is important for the morphology and permeability of the parasitophorous vacuole in Toxoplasma gondii GRA47 对弓形虫寄生泡的形态和渗透性非常重要。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-01 DOI: 10.1016/j.ijpara.2024.06.003

Establishing an intact intracellular parasitophorous vacuole (PV) that enables efficient nutrient uptake and protein trafficking is essential for the survival and proliferation of Toxoplasma gondii. Although the PV membrane (PVM)-localized dense granule protein 17 (GRA17) and GRA23 mediate the permeability of the PVM to small molecules, including nutrient uptake and excretion of metabolic by-products, the molecular mechanism by which T. gondii acquires nutrients remains unclear. In this study, we showed that the secreted protein GRA47 contributed to normal PV morphology, PVM permeability to small molecules, growth, and virulence in T. gondii. Co-immunoprecipitation analysis demonstrated potential interaction of GRA47 with GRA72, and the loss of GRA72 affected PV morphology, parasite growth and infectivity. To investigate the biological relationship among GRA47, GRA72, GRA17 and GRA23, attempts were made to construct strains with double gene deletion and overexpressing strains. Only Δgra23Δgra72 was successfully constructed. This strain exhibited a significant increase in the proportion of aberrant PVs compared with the Δgra23 strain. Overexpressing one of the three related GRAs partially rescued PVs with aberrant morphology in Δgra47, Δgra72 and Δgra17, while the expression of the Plasmodium falciparum PVM protein PfExp2, an ortholog of GRA17 and GRA23, fully rescued the PV morphological defect in all three Δgra strains. These results suggest that these GRA proteins may not be functionally redundant but rather work in different ways to regulate nutrient acquisition. These findings highlight the versatility of the nutrient uptake mechanisms in T. gondii, which may contribute to the parasite’s remarkable ability to grow in different cellular niches in a very broad range of hosts.

建立一个完整的细胞内寄生虫液泡(PV),使其能够有效地摄取营养物质和贩运蛋白质,对于弓形虫的生存和增殖至关重要。虽然PV膜(PVM)定位的致密颗粒蛋白17(GRA17)和GRA23介导了PVM对小分子的通透性,包括营养物质的摄取和代谢副产物的排泄,但弓形虫获取营养物质的分子机制仍不清楚。在这项研究中,我们发现分泌蛋白 GRA47 对淋病双球菌正常的 PV 形态、PVM 对小分子的通透性、生长和毒力都有贡献。共免疫沉淀分析表明 GRA47 与 GRA72 有潜在的相互作用,而 GRA72 的缺失会影响 PV 形态、寄生虫的生长和感染性。为了研究 GRA47、GRA72、GRA17 和 GRA23 之间的生物学关系,尝试构建了双基因缺失株和过表达株。结果只成功构建了Δgra23Δgra72。与 Δgra23 株系相比,该株系的异常 PV 比例明显增加。过表达三个相关 GRA 中的一个可部分修复 Δgra47、Δgra72 和 Δgra17 中形态异常的 PV,而表达恶性疟原虫 PVM 蛋白 PfExp2(GRA17 和 GRA23 的直系同源物)可完全修复所有三个 Δgra 株系中的 PV 形态缺陷。这些结果表明,这三种 GRA 蛋白可能并非功能冗余,而是以不同的方式调节营养物质的获取。这些发现凸显了淋球菌营养摄取机制的多样性,这可能是淋球菌能够在多种宿主的不同细胞环境中生长的重要原因。
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引用次数: 0
What shapes a microbiome? Differences in bacterial communities associated with helminth-amphipod interactions. 是什么塑造了微生物群落?与蠕虫-片脚类动物相互作用相关的细菌群落差异
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-08-30 DOI: 10.1016/j.ijpara.2024.08.005
Célia Koellsch, Robert Poulin, Priscila M Salloum

The fast technological advances of molecular tools have enabled us to uncover a new dimension hidden within parasites and their hosts: their microbiomes. Increasingly, parasitologists characterise host microbiome changes in the face of parasitic infections, revealing the potential of these microscopic fast-evolving entities to influence host-parasite interactions. However, most of the changes in host microbiomes seem to depend on the host and parasite species in question. Furthermore, we should understand the relative role of parasitic infections as microbiome modulators when compared with other microbiome-impacting factors (e.g., host size, age, sex). Here, we characterised the microbiome of a single intermediate host species infected by two parasites belonging to different phyla: the acanthocephalan Plagiorhynchus allisonae and a dilepidid cestode, both infecting Transorchestia serrulata amphipods collected simultaneously from the same locality. We used the v4 hypervariable region of the 16S rRNA prokaryotic gene to identify the hemolymph bacterial community of uninfected, acanthocephalan-infected, and cestode-infected amphipods, as well as the bacteria in the amphipods' immediate environment and in the parasites infecting them. Our results show that parasitic infections were more strongly associated with differences in host bacterial community richness than amphipod size, presence of amphipod eggs in female amphipods, and even parasite load. Amphipods infected by acanthocephalans had the most divergent bacterial community, with a marked decrease in alpha diversity compared with cestode-infected and uninfected hosts. In accordance with the species-specific nature of microbiome changes in parasitic infections, we found unique microbial taxa associating with hosts infected by each parasite species, as well as taxa only shared between a parasite species and their infected hosts. However, there were some bacterial taxa detected in all parasitised amphipods (regardless of the parasite species), but not in uninfected amphipods or the environment. We propose that shared bacteria associated with all hosts parasitised by distantly related helminths may be important either in helping host defences or parasites' success, and could thus interact with host-parasite evolution.

分子工具技术的飞速发展使我们能够发现隐藏在寄生虫及其宿主体内的一个新维度:寄生虫微生物组。越来越多的寄生虫学家描述了寄生虫感染时宿主微生物组的变化,揭示了这些快速进化的微观实体影响宿主-寄生虫相互作用的潜力。然而,宿主微生物组的大多数变化似乎取决于宿主和寄生虫的种类。此外,与其他影响微生物组的因素(如宿主大小、年龄、性别)相比,我们应该了解寄生虫感染作为微生物组调节剂的相对作用。在这里,我们描述了被属于不同门类的两种寄生虫感染的单一中间宿主的微生物组特征:棘头蚤(Plagiorhynchus allisonae)和双尾蚤(dilepidid cestode),这两种寄生虫都感染了从同一地点同时采集的Transorchestia serrulata片脚类动物。我们利用 16S rRNA 原核基因的 v4 超变区鉴定了未感染、感染棘头蚴和感染绦虫的片脚类动物的血淋巴细菌群落,以及片脚类动物周围环境和感染片脚类动物的寄生虫中的细菌。我们的研究结果表明,寄生虫感染与寄主细菌群落丰富度的差异有更密切的关系,而与片脚类动物的大小、雌性片脚类动物体内片脚类动物卵的存在,甚至寄生虫量的差异都没有关系。被棘尾虫感染的片脚类动物的细菌群落差异最大,与绦虫感染和未感染的宿主相比,α多样性明显下降。根据寄生虫感染时微生物群落变化的物种特异性,我们发现了与每种寄生虫感染的宿主相关的独特微生物类群,以及仅在寄生虫物种和感染宿主之间共享的类群。然而,在所有被寄生的片脚类动物(无论寄生虫种类)中都检测到了一些细菌类群,但在未感染的片脚类动物或环境中却没有检测到。我们认为,与所有被远缘蠕虫寄生的宿主相关的共有细菌可能在帮助宿主防御或寄生虫成功寄生方面具有重要作用,因此可能与宿主-寄生虫的进化相互影响。
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引用次数: 0
Bayesian estimation of the sensitivity and specificity of coprological and serological diagnostic tests for the detection of Ascaris suum infection on pig farms 用贝叶斯法估算猪场蛔虫感染检测中桡骨学和血清学诊断测试的灵敏度和特异性
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-08-01 DOI: 10.1016/j.ijpara.2024.04.010

Coprological and serological diagnostic tests were compared to define the status of a pig farm with regard to Ascaris suum. On each of the 100 farms in France visited for the study, 10 blood samples were taken from pigs at the end of fattening (at least 22 weeks old) and 20 to 30 faecal samples were taken, depending on the category of animals present on the farm (10 sows, 10 piglets aged 10 to 12 weeks and 10 pigs at the end of fattening, aged at least 22 weeks). A SERASCA® ELISA test (Laboratory of Parasitology, Ghent University) was performed on each blood sample (cut-off 0.5) and a coprological analysis on each faecal sample. A Bayesian approach was used to estimate the sensitivity and specificity of the coprological and serological tests. A farm was considered positive if at least one A. suum egg was observed in the faecal samples. With regard to the serological test, various hypotheses were tested in order to define the number of seropositive animals required to consider a farm positive for A. suum. The coprological test has very good specificity in the search for A. suum, whether 20 or 30 samples are taken per farm. However, even with an increase in the number of samples, the sensitivity of this diagnostic approach is very low (less than 30%). On the other hand, the serological diagnostic method, which consists of taking blood samples from 10 animals at the end of fattening, has good sensitivity and seems better suited to defining the status of a farm with regard to A. suum, provided that a farm is considered seropositive only if two out of 10 samples are positive.

通过比较弧菌和血清学诊断测试来确定养猪场的蛔虫状况。在法国考察的 100 个猪场中,每个猪场从育肥末期(至少 22 周龄)的猪身上采集 10 份血液样本,并根据猪场的动物类别(10 头母猪、10 头 10-12 周龄的仔猪和 10 头育肥末期(至少 22 周龄)的猪)采集 20-30 份粪便样本。对每份血液样本进行 SERASCA® 酶联免疫吸附试验(根特大学寄生虫学实验室)(临界值为 0.5),对每份粪便样本进行粪便分析。贝叶斯法用于估算桡骨学和血清学检测的灵敏度和特异性。如果在粪便样本中观察到至少一个鼠卵,则该农场被视为阳性。在血清学检测方面,对各种假设进行了测试,以确定将一个猪场判定为鼠疫阳性所需的血清阳性动物数量。无论每个猪场采集 20 个样本还是 30 个样本,粪便检测对猪甲虫的检测都有很好的特异性。然而,即使样本数量增加,这种诊断方法的灵敏度也非常低(低于 30%)。另一方面,血清学诊断方法是在育肥结束时抽取 10 头牲畜的血液样本,灵敏度较高,似乎更适于确定猪场的猪蓝耳病状况,前提是 10 个样本中有 2 个呈阳性,猪场才会被视为血清阳性。
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引用次数: 0
Exploring the second intermediate hosts and morphology of human- and cat-specific Opisthorchis viverrini-like populations 探索人猫特异性 Opisthorchis viverrini-like 种群的第二中间宿主和形态。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-08-01 DOI: 10.1016/j.ijpara.2024.04.006

Infection by the zoonotic fish-borne trematode, Opisthorchis viverrini, remains a crucial health issue in Thailand and neighboring countries. Recently, molecular analysis revealed two populations of putative O. viverrini: one found primarily in human hosts (“human-specific” population) and the other primarily in cats (“cat-specific” population). It is unclear how the infective stages (metacercariae) of these different populations circulate among definitive and reservoir hosts in nature. To gain an insight into this, mitochondrial cox1 and nad1 gene sequences of metacercariae from fish intermediate hosts were examined. None of 192 metacercariae from cyprinid fish in Lao PDR and Thailand had sequences typical of “cat-specific” O. viverrini, suggesting that cyprinid fish are not the main second intermediate hosts of this population. Interestingly, all 20 O. viverrini-like metacercariae from snakehead fish (Channa striata) shared 99.51–100% sequence identity with eggs from cats naturally infected in a previous study. Hence, we propose a modification of the known transmission dynamics of O. viverrini: consumption of metacercariae within snakehead fish provides another pathway for cats and (occasionally) humans to acquire infection. We also performed morphological comparisons of eggs, metacercariae, and adult flukes (raised in hamsters) of both Opisthorchis populations. The “cat-specific” population has eggs that are narrower and adults that are shorter and wider than in the human-specific population. The metacercaria of the “cat-specific” population is elliptical, while that of the “human-specific” population is oval, occasionally rounded. Our results confirmed that O. viverrini-like metacercariae from snakehead fish are the infective stages of the “cat-specific” fluke. This provides a new insight into the dissemination and transmission of each population in the second intermediate host. The identity of the cat-specific population is discussed.

在泰国及其邻国,人畜共患的鱼源性颤形虫(Opisthorchis viverrini)感染仍然是一个重要的健康问题。最近,分子分析发现了两种假定的 O. viverrini 群体:一种主要存在于人类宿主中("人类特异性 "群体),另一种主要存在于猫("猫特异性 "群体)中。目前还不清楚这些不同种群的感染阶段(元孢子虫)如何在自然界中的确定宿主和贮存宿主之间循环。为了深入了解这一问题,我们研究了鱼类中间宿主的线粒体 cox1 和 nad1 基因序列。老挝人民民主共和国和泰国鲤科鱼类的 192 个盘尾丝虫中没有一个具有 "猫特异性 "O. viverrini 的典型序列,这表明鲤科鱼类不是该种群的主要第二中间宿主。有趣的是,乌鳢(Channa striata)的所有 20 个 O. viverrini-like metaceriae 都与之前研究中自然感染的猫卵有 99.51-100% 的序列相同性。因此,我们提出了一种对已知 O. viverrini 传播动态的修正:食用乌鳢体内的类盘虫为猫和(偶尔)人类提供了另一种感染途径。我们还对两个Opisthorchis种群的卵、盘尾丝蚴和成虫(在仓鼠体内饲养)进行了形态学比较。与人类特异性种群相比,"猫特异性 "种群的虫卵更窄,成虫更短更宽。猫特异性 "种群的包虫呈椭圆形,而 "人类特异性 "种群的包虫呈椭圆形,偶尔也有圆形。我们的研究结果证实,来自乌鳢的类O. viverrini metaceriae是 "猫特异性 "吸虫的感染阶段。这为我们了解每个种群在第二中间宿主中的传播情况提供了新的视角。本文还讨论了 "猫特异性 "种群的身份。
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引用次数: 0
Novel CRISPR-Cas-powered pen-side test for East Coast fever 新型CRISPR-Cas驱动的东海岸热笔侧测试
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-08-01 DOI: 10.1016/j.ijpara.2024.04.009

Theileria parva causes East Coast fever (ECF), one of the most important and lethal tick-borne diseases of cattle in sub-Saharan Africa. ECF is a considerable burden to the livestock industry, causing annual losses exceeding US $300 million. Currently, diagnosis of T. parva infections relies mainly on clinical signs, serology, and microscopic identification of parasites in either blood or lymph fluid samples. However, some of these tests might not indicate ongoing infection and they all lack the sensitivity to detect low-level infections. Molecular tests such as nested and quantitative PCR assays offer high sensitivity for detection of T. parva. However, these tests remain highly complex technologies that are impractical to use in resource-limited settings where economic losses due to the disease have the most significant impact. A field-deployable, point-of-care test will be of significant value in the treatment and control of ECF in endemic areas. For this purpose, we have developed a CRISPR-Cas12a-based pen-side tool that can sensitively and specifically detect T. parva based on the p104 gene. We describe a streamlined, field-applicable diagnostic tool comprising a 20 min recombinase polymerase amplification (RPA) reaction followed by a 60 min CRISPR-Cas12a reaction using a FAM/Biotin lateral flow strip readout. We tested two different RPA primer pairs and four different CRISPR-RNAs (crRNAs). The p104-based assay displayed high sensitivity, detecting as low as one infected lymphocyte per three microliters of blood and universally detecting eight different T. parva strains without detecting DNA from other Theileria spp. such as Theileria mutans and Theileria lestoquardi. This work opens the way for a field-applicable diagnostic tool for the sensitive point-of-care early diagnosis of T. parva infections in cattle.

东海岸热(ECF)是撒哈拉以南非洲地区最重要、最致命的牛蜱媒疾病之一,由副疟原虫(Theileria parva)引起。东海岸热是畜牧业的沉重负担,每年造成的损失超过 3 亿美元。目前,T. parva 感染的诊断主要依靠临床症状、血清学和血液或淋巴液样本中寄生虫的显微鉴定。然而,其中一些检测方法可能无法提示正在发生的感染,而且它们都缺乏检测低水平感染的灵敏度。巢式 PCR 和定量 PCR 等分子检测方法对 Parva T. 的检测灵敏度较高。然而,这些检测方法仍然是非常复杂的技术,在资源有限的环境中使用并不现实,因为这种疾病造成的经济损失对这些环境的影响最大。一种可现场部署的护理点检测方法将对流行地区的ECF治疗和控制具有重要价值。为此,我们开发了一种基于 CRISPR-Cas12a 的笔端工具,可以根据 p104 基因灵敏、特异地检测副猪嗜血杆菌。我们描述了一种简化的现场适用诊断工具,包括 20 分钟的重组酶聚合酶扩增(RPA)反应和 60 分钟的 CRISPR-Cas12a 反应,使用 FAM/Biotin 侧流条带读数。我们测试了两种不同的 RPA 引物对和四种不同的 CRISPR-RNA (crRNA)。基于 p104 的检测方法显示出极高的灵敏度,每三微升血液可检测到一个受感染的淋巴细胞,并能普遍检测到八种不同的副猪嗜血杆菌菌株,而不会检测到变异副猪嗜血杆菌(Theileria mutans)和雌性副猪嗜血杆菌(Theileria lestoquardi)等其他副猪嗜血杆菌属的 DNA。这项工作为现场适用的诊断工具开辟了道路,可用于敏感的牛副嗜血杆菌感染的护理点早期诊断。
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引用次数: 0
New member of Plasmodium (Vinckeia) and Plasmodium cyclopsi discovered in bats in Sierra Leone – nuclear sequence and complete mitochondrial genome analyses 在塞拉利昂的蝙蝠中发现疟原虫(Vinckeia)和环状疟原虫的新成员--核序列和完整线粒体基因组分析。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-08-01 DOI: 10.1016/j.ijpara.2024.05.002

Malaria remains the most important arthropod-borne infectious disease globally. The causative agent, Plasmodium, is a unicellular eukaryote that develops inside red blood cells. Identifying new Plasmodium parasite species that infect mammalian hosts can shed light on the complex evolution and diversity of malaria parasites. Bats feature a high diversity of microorganisms including seven separate genera of malarial parasites. Three species of Plasmodium have been reported so far, for which scarce reports exist. Here we present data from an investigation of Plasmodium infections in bats in the western Guinean lowland forest in Sierra Leone. We discovered a new Plasmodium parasite in the horseshoe bat Rhinolophus landeri. Plasmodium cyclopsi infections in a member of leaf-nosed bats, Doryrhina cyclops, exhibited a high prevalence of 100%. Phylogenetic analysis of complete mitochondrial genomes and nine nuclear markers recovered a close relationship between P. cyclopsi and the new Plasmodium parasite with the rodent species Plasmodium berghei, a widely used in vivo model to study malaria in humans. The data suggests that the “rodent/bat” Plasmodium (Vinckeia) clade represents a diverse group of malarial parasites that would likely expand with a systematic sampling of small mammals in tropical Africa. Identifying the bat Plasmodium repertoire is central to our understanding of the evolution of Plasmodium parasites in mammals.

疟疾仍然是全球最重要的节肢动物传染病。致病菌疟原虫是一种在红细胞内发育的单细胞真核生物。鉴定感染哺乳动物宿主的疟原虫新物种可以揭示疟原虫复杂的进化和多样性。蝙蝠体内的微生物种类繁多,包括七个不同的疟原虫属。迄今为止,已报道的疟原虫有 3 种,但相关报道很少。在这里,我们展示了塞拉利昂几内亚西部低地森林中蝙蝠感染疟原虫的调查数据。我们在马蹄蝠 Rhinolophus landeri 身上发现了一种新的疟原虫寄生体。在叶鼻蝠的一个成员 Doryrhina cyclops 中,环状疟原虫的感染率高达 100%。对完整线粒体基因组和九个核标记的系统发育分析表明,环眼疟原虫和新疟原虫与啮齿类动物疟原虫之间的关系密切,而啮齿类动物疟原虫是研究人类疟疾的一个广泛使用的体内模型。这些数据表明,"啮齿类/蝙蝠 "疟原虫(Vinckeia)支系代表了一个多样化的疟疾寄生虫群体,随着对非洲热带小型哺乳动物进行系统取样,该群体很可能会扩大。确定蝙蝠疟原虫谱系对于我们了解哺乳动物中疟原虫寄生虫的进化至关重要。
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引用次数: 0
Pathogen-associated molecular patterns (PAMPs) derived from Leishmania and bacteria increase gene expression of antimicrobial peptides and gut surface proteins in sand flies 来自利什曼原虫和细菌的病原体相关分子模式(PAMPs)会增加沙蝇体内抗菌肽和肠道表面蛋白的基因表达。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-08-01 DOI: 10.1016/j.ijpara.2024.04.005

The interaction between pathogens and vectors’ physiology can impact parasite transmission. Studying this interaction at the molecular level can help in developing control strategies. We study leishmaniases, diseases caused by Leishmania parasites transmitted by sand fly vectors, posing a significant global public health concern. Lipophosphoglycan (LPG), the major surface glycoconjugate of Leishmania, has been described to have several roles throughout the parasite’s life cycle, both in the insect and vertebrate hosts. In addition, the sand fly midgut possesses a rich microbiota expressing lipopolysaccharides (LPS). However, the effect of LPG and LPS on the gene expression of sand fly midgut proteins or immunity effectors has not yet been documented. We experimentally fed Lutzomyia longipalpis and Phlebotomus papatasi sand flies with blood containing purified LPG from Leishmania infantum, Leishmania major, or LPS from Escherichia coli. The effect on the expression of genes encoding gut proteins galectin and mucin, digestive enzymes trypsin and chymotrypsin, and antimicrobial peptides (AMPs) attacin and defensins was assessed by quantitative PCR (qPCR). The gene expression of a mucin-like protein in L. longipalpis was increased by L. infantum LPG and E. coli LPS. The gene expression of a galectin was increased in L. longipalpis by L. major LPG, and in P. papatasi by E. coli LPS. Nevertheless, the gene expression of trypsins and chymotrypsins did not significantly change. On the other hand, both L. infantum and L. major LPG significantly enhanced expression of the AMP attacin in both sand fly species and defensin in L. longipalpis. In addition, E. coli LPS increased the expression of attacin and defensin in L. longipalpis. Our study showed that Leishmania LPG and E. coli LPS differentially modulate the expression of sand fly genes involved in gut maintenance and defence. This suggests that the glycoconjugates from microbiota or Leishmania may increase the vector’s immune response and the gene expression of a gut coating protein in a permissive vector.

病原体与病媒生理之间的相互作用会影响寄生虫的传播。从分子水平研究这种相互作用有助于制定控制策略。我们研究的利什曼病是由沙蝇媒介传播的利什曼寄生虫引起的疾病,是全球公共卫生的重大问题。脂磷聚糖(LPG)是利什曼原虫的主要表面糖类共轭物,在寄生虫的整个生命周期中,无论是在昆虫宿主还是在脊椎动物宿主体内,都发挥着多种作用。此外,沙蝇中肠拥有丰富的表达脂多糖(LPS)的微生物群。然而,LPG 和 LPS 对沙蝇中肠蛋白或免疫效应因子基因表达的影响尚未被记录。我们用含有纯化的幼年利什曼原虫、大利什曼原虫LPG或大肠杆菌LPS的血液喂养Lutzomyia longipalpis和Phlebotomus papatasi沙蝇。通过定量聚合酶链反应(qPCR)评估了对编码肠道蛋白galectin和粘蛋白、消化酶胰蛋白酶和糜蛋白酶以及抗菌肽(AMPs)attacin和防御素(defensins)的基因表达的影响。婴儿痢疾杆菌 LPG 和大肠杆菌 LPS 增加了长舌蝇体内一种粘蛋白样蛋白的基因表达。大肠杆菌 LPG 和大肠杆菌 LPS 均增加了长尾蝇中一种半凝集素的基因表达量。然而,胰蛋白酶和糜蛋白酶的基因表达量没有显著变化。另一方面,L. infantum 和 L. major LPG 能显著提高两种沙蝇的 AMP attacin 和 L. longipalpis 的 defensin 的表达。此外,大肠杆菌 LPS 也能提高长爪蝇中的attacin 和 defensin 的表达。我们的研究表明,利什曼病菌 LPG 和大肠杆菌 LPS 对参与肠道维护和防御的沙蝇基因的表达有不同的调节作用。这表明,微生物群或利什曼病菌的糖类共轭物可能会增加载体的免疫反应,并增加许可载体肠道涂层蛋白的基因表达。
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引用次数: 0
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International journal for parasitology
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