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Optimized plasmid loading of human erythrocytes for Plasmodium falciparum DNA transfections 用于恶性疟原虫 DNA 转染的人红细胞质粒装载优化。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-10-01 DOI: 10.1016/j.ijpara.2024.04.011
In vitro modification of Plasmodium falciparum genes is the cornerstone of basic and translational malaria research. Achieved through DNA transfection, these modifications may entail altering protein sequence or abundance. Such experiments are critical for defining the molecular mechanisms of key parasite phenotypes and for validation of drug and vaccine targets. Despite its importance, successful transfection remains difficult and is a resource-intensive, rate-limiting step in P. falciparum research. Here, we report that inefficient loading of plasmid into erythrocytes limits transfection efficacy with commonly used electroporation methods. As these methods also require expensive instrumentation and consumables that are not broadly available, we explored a simpler method based on plasmid loading through hypotonic lysis and resealing of erythrocytes. We used parasite expression of a sensitive NanoLuc reporter for rapid evaluation and optimization of each step. Hypotonic buffer composition, resealing buffer volume and composition, and subsequent incubation affected plasmid retention and successful transfection. While ATP was critical for erythrocyte resealing, addition of Ca++ or glutathione did not improve transfection efficiency, with increasing Ca++ concentrations proving detrimental to outcomes. Compared with either the standard electroporation method or a previously reported hypotonic loading protocol, the optimized method yields greater plasmid loading and higher expression of the NanoLuc reporter 48 h after transfection. It also produced significantly faster outgrowth of parasites in transfections utilizing either episomal expression or CRISPR-Cas9 mediated integration. This new method produces higher P. falciparum transfection efficiency, reduces resource requirements and should accelerate molecular studies of malaria drug and vaccine targets.
恶性疟原虫基因的体外改造是疟疾基础研究和转化研究的基石。通过 DNA 转染,这些修饰可能会改变蛋白质序列或丰度。此类实验对于确定关键寄生虫表型的分子机制以及验证药物和疫苗靶点至关重要。尽管转染很重要,但成功转染仍然很困难,是恶性疟原虫研究中一个资源密集型的限制性步骤。在此,我们报告了质粒在红细胞中的低效负载限制了常用电穿孔方法的转染效果。由于这些方法还需要昂贵的仪器和耗材,而这些仪器和耗材并不能广泛使用,因此我们探索了一种更简单的方法,即通过低渗裂解红细胞并重新封口来装载质粒。我们利用寄生虫表达敏感的 NanoLuc 报告器来快速评估和优化每个步骤。低渗缓冲液的成分、再封闭缓冲液的容量和成分以及随后的培养都会影响质粒的保留和成功转染。虽然 ATP 对红细胞再封闭至关重要,但添加 Ca++ 或谷胱甘肽并不能提高转染效率,Ca++ 浓度的增加对转染结果不利。与标准电穿孔方法或之前报道的低渗装载方案相比,优化方法能在转染 48 小时后产生更多的质粒装载和更高的 NanoLuc 报告基因表达。在利用外显子表达或 CRISPR-Cas9 介导的整合进行转染时,它还能明显加快寄生虫的生长速度。这种新方法可提高恶性疟原虫的转染效率,减少资源需求,并可加速疟疾药物和疫苗靶标的分子研究。
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引用次数: 0
Tick salivary proteome and lipidome with low glycan content correlate with allergic type reactions in the zebrafish model 低糖含量的蜱唾液蛋白质组和脂质组与斑马鱼模型中的过敏型反应相关。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-10-01 DOI: 10.1016/j.ijpara.2024.07.002
Ticks, as hematophagous ectoparasites, can manipulate host immune and metabolic processes, causing tick-borne allergies such as α-Gal syndrome (AGS). Glycolipids with bound galactose-alpha-1–3-galactose (α-Gal) are potential allergenic molecules associated with AGS. Nevertheless, proteins and lipids lacking α-Gal modifications may contribute to tick salivary allergies and be linked to AGS. In this study, we characterized the effect of deglycosylated tick salivary proteins without lipids on treated zebrafish fed with dog food formulated with mammalian (beef, lamb, pork) meat by quantitative proteomics analysis of intestinal samples. The characterization and functional annotations of tick salivary lipids with low representation of glycolipids was conducted using a lipidomics approach. Results showed a significant effect of treatment with saliva and saliva deglycosylated protein fraction on zebrafish abnormal or no feeding (p < 0.005). Treatment with this fraction affected multiple metabolic pathways, defense responses to pathogens and protein metabolism, which correlated with abnormal or no feeding. Lipidomics analysis identified 23 lipid classes with low representation of glycolipids (0.70% of identified lipids). The lipid class with highest representation was phosphatidylcholine (PC; 26.66%) and for glycolipids it corresponded to diacylglycerol (DG; 0.48%). Qualitative analysis of PC antibodies revealed that individuals bitten by ticks were more likely to produce PC-IgG antibodies (p < 0.001). DG levels were significantly higher in tick salivary glands (p < 0.05) compared with tick saliva and salivary fractions. The α-Gal content was higher in tick saliva than in deglycosylated saliva and lipid fractions. These results support a possible role for tick salivary proteins and lipids without α-Gal modifications in AGS.
蜱虫作为食血的体外寄生虫,可以操纵宿主的免疫和代谢过程,导致蜱传过敏症,如α-Gal综合征(AGS)。与半乳糖-α-1-3-半乳糖(α-Gal)结合的糖脂是与 AGS 相关的潜在过敏原分子。然而,缺乏α-Gal修饰的蛋白质和脂质也可能导致蜱唾液过敏,并与AGS有关。在这项研究中,我们通过对肠道样本进行定量蛋白质组学分析,确定了不含脂质的脱糖蜱唾液蛋白对用哺乳动物(牛肉、羊肉、猪肉)肉配制的狗粮喂养的斑马鱼的影响。利用脂质组学方法对糖脂含量较低的蜱唾液脂质进行了表征和功能注释。结果表明,用唾液和唾液脱糖蛋白部分处理斑马鱼异常或不摄食有明显影响(p < 0.005)。用唾液和唾液脱糖蛋白处理斑马鱼会影响多种代谢途径、对病原体的防御反应和蛋白质代谢,这与斑马鱼摄食异常或不摄食有关。脂质组学分析确定了 23 类脂质,其中糖脂的代表性较低(占所确定脂质的 0.70%)。代表性最高的脂类是磷脂酰胆碱(PC;26.66%),糖脂类则是二酰甘油(DG;0.48%)。PC 抗体的定性分析显示,被蜱虫叮咬的个体更容易产生 PC-IgG 抗体(p < 0.001)。与蜱唾液和唾液组分相比,蜱唾液腺中的 DG 含量明显更高(p < 0.05)。蜱唾液中的α-Gal含量高于脱糖唾液和脂质部分。这些结果支持了蜱唾液蛋白质和脂质在 AGS 中可能起的作用。
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引用次数: 0
Detection of Toxoplasma gondii in wild bivalves from the Kerguelen and Galapagos archipelagos: influence of proximity to cat populations, exposure to marine currents and kelp density 在凯尔盖朗群岛和加拉帕戈斯群岛的野生双壳贝中检测到弓形虫:与猫群的距离、暴露于海流和海藻密度的影响
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-10-01 DOI: 10.1016/j.ijpara.2024.06.001
Oocysts of the protozoan Toxoplasma gondii are found in felid feces and can be washed into coastal waters, where they persist for months, attaching to algae and accumulating in invertebrates. We used wild bivalves to assess contamination of coastal waters of the Kerguelen and Galapagos archipelagos by this zoonotic parasite. Additionally, we leveraged the contrasting situations of these archipelagos to identify some potential drivers of contamination. In the Galapagos, with a cat density reaching 142 per km2, 15.38% of the sampled oysters (Saccostrea palmula) tested positive for T. gondii by quantitative real-time PCR (qPCR) (n = 260), and positive samples were found in all eight sampling sites. In Kerguelen, with 1-3 cats per km2, 40.83% of 120 tested mussels (Mytilus edulis platensis) were positive, and positive samples were found in four out of the five sampling sites. These findings provide evidence of T. gondii contamination in the coastal waters of these archipelagos. Furthermore, T. gondii-positive bivalves were found on islands located 20 km away (Galapagos) and 5 km away (Kerguelen) from the nearest cat population, indicating that T. gondii oocysts can disperse through waterborne mechanisms over several kilometers from their initial deposition site. In the Galapagos, where runoff is infrequent and all sites are exposed to currents, the prevalence of qPCR-positive bivalves did not show significant variations between sites (p = 0.107). In Kerguelen where runoff is frequent and site exposure variable, the prevalence varied significantly (p < 0.001). The detection of T. gondii in Kerguelen mussels was significantly correlated with the site exposure to currents (odds ratio (OR) 60.2, p < 0.001) and the on-site density of giant kelp forests (OR 2.624, p < 0.001). This suggests that bivalves can be contaminated not only by oocysts transported by currents but also by consuming marine aggregates containing oocysts that tend to form in kelp forests.
原生动物弓形虫的卵囊存在于猫科动物的粪便中,会被冲入沿岸水域,并在那里存留数月,附着在藻类上,并在无脊椎动物体内积累。我们利用野生双壳贝类来评估这种人畜共患病寄生虫对凯尔盖朗群岛和加拉帕戈斯群岛沿岸水域的污染情况。此外,我们还利用这两个群岛截然不同的情况,确定了一些潜在的污染驱动因素。在加拉帕戈斯群岛,猫的密度达到每平方公里 142 只,通过实时定量 PCR(qPCR)检测,15.38% 的采样牡蛎(Saccostrea palmula)的淋病双球菌检测呈阳性(n = 260),在所有八个采样点都发现了阳性样本。在凯尔盖朗(Kerguelen),每平方公里有 1-3 只猫,在 120 个检测的贻贝(Mytilus edulis platensis)中,有 40.83% 呈阳性,在五个采样点中的四个采样点都发现了阳性样本。这些发现提供了这些群岛沿海水域受到淋病双球菌污染的证据。此外,在距离最近的猫群 20 千米(加拉帕戈斯群岛)和 5 千米(凯尔盖朗岛)以外的岛屿上也发现了 T. gondii 阳性双壳贝,这表明 T. gondii 卵囊可以通过水传播机制从最初的沉积地点扩散到几千米以外的地方。在加拉帕戈斯群岛,径流并不频繁,而且所有地点都暴露在水流中,qPCR 阳性双壳贝的流行率在不同地点之间并无显著差异(p = 0.107)。在凯尔盖朗岛,由于径流频繁,各地点的水流情况各不相同,因此双壳类动物的发病率差异很大(p = 0.001)。在凯尔盖朗贻贝中检测到的淋病双球菌与现场暴露于水流的情况(几率比(OR)60.2,p <0.001)和现场巨型海藻林的密度(OR 2.624,p <0.001)有明显的相关性。这表明,双壳类动物不仅会受到由水流传播的卵囊的污染,也会因食用海藻林中往往会形成的含有卵囊的海洋聚集体而受到污染。
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引用次数: 0
Unraveling new players in helminth pathology: extracellular vesicles from Fasciola hepatica and Dicrocoelium dendriticum exert different effects on hepatic stellate cells and hepatocytes 揭示蠕虫病理学中的新角色:来自肝脏法氏囊和树枝状微囊藻的细胞外囊泡对肝星状细胞和肝细胞产生不同影响。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-10-01 DOI: 10.1016/j.ijpara.2024.06.002
Fasciola hepatica and Dicrocoelium dendriticum are parasitic trematodes residing in the bile ducts of mammalian hosts, causing, in some cases, impairment of liver function and hepatic fibrosis. Previous studies have shown that extracellular vesicles released by F. hepatica (FhEVs) and D. dendriticum (DdEVs) induce a distinct phenotype in human macrophages, but there is limited information on the effect of parasitic EVs on liver cells, which interact directly with the worms in natural infections. In this study, we isolated FhEVs and DdEVs by size exclusion chromatography and labeled them with a lipophilic fluorescent dye to analyze their uptake by human hepatic stellate cells (HSC) and hepatocytes, important cell types in liver pathology, using synthetic liposomes as internal labeling and uptake control. We analyzed EV uptake and the proteome profiles after the treatment with EVs for both cell types. Our results reveal that EVs establish unique and specific interactions with stellate cells and hepatocytes, suggesting a different role of EVs derived from each parasite, depending on the migration route to reach their final niche. FhEVs have a cytostatic effect on HSCs, but induce the extracellular matrix secretion and elicit anti-inflammatory responses in hepatocytes. DdEVs have a more potent anti-proliferative effect than FhEVs and trigger a global inflammatory response, increasing the levels of NF-κB and other inflammatory mediators in both cell types. These interactions may have a major influence on the progression of the disease, serving to generate conditions that may favor the establishment of the helminths in the host.
Fasciola hepatica 和 Dicrocoelium dendriticum 是寄生在哺乳动物宿主胆管中的吸虫,在某些情况下会导致肝功能受损和肝纤维化。以前的研究表明,肝吸虫(FhEVs)和树枝状吸虫(DdEVs)释放的胞外囊泡会诱导人类巨噬细胞出现不同的表型,但关于寄生EVs对肝细胞影响的信息却很有限,因为在自然感染中,肝细胞会直接与虫体发生相互作用。在这项研究中,我们通过尺寸排阻色谱法分离了FhEVs和DdEVs,并用亲脂性荧光染料对其进行标记,以合成脂质体作为内部标记和摄取对照,分析肝脏病理学中重要的细胞类型--人肝星状细胞(HSC)和肝细胞对它们的摄取情况。我们分析了这两种细胞对EVs的吸收情况以及EVs处理后的蛋白质组图谱。我们的研究结果表明,EVs 与星状细胞和肝细胞建立了独特的特异性相互作用,这表明每种寄生虫产生的 EVs 都有不同的作用,这取决于它们到达最终龛位的迁移路线。FhEVs对造血干细胞有细胞抑制作用,但能诱导细胞外基质分泌并引起肝细胞的抗炎反应。与 FhEVs 相比,DdEVs 具有更强的抗增殖作用,并引发整体炎症反应,提高两种细胞中 NF-κB 和其他炎症介质的水平。这些相互作用可能会对疾病的进展产生重大影响,从而产生有利于蠕虫在宿主体内繁殖的条件。
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引用次数: 0
Parasite airlines: mapping the distribution and transmission of avian blood parasites in migratory birds 寄生虫航线:绘制候鸟血液寄生虫的分布和传播图。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-10-01 DOI: 10.1016/j.ijpara.2024.06.004
During their journeys, migratory birds encounter a wider range of parasites than residents, transporting them over vast distances. While some parasites are widely distributed, transmission is not inevitable and depends on the presence of competent arthropod vectors as well as parasite compatibility with native bird species. Distinguishing between parasite distribution and transmission areas is crucial for monitoring and assessing risks to native bird species, as distribution areas, with the appropriate conditions, could become potential transmission areas. In this study, blood samples from 455 reed-living birds of the genera Acrocephalus, Locustella, and Emberiza, collected in the nature reserve “Die Reit” in Hamburg, Germany were screened, targeting haemosporidian parasites, trypanosomes, and filarioid nematodes. Determination of migratory bird age was employed to ascertain the transmission area of the detected parasites. Transmission areas were determined, based on information provided by resident and juvenile birds as well as findings in competent vectors. Long-distance migratory birds of the genus Acrocephalus showed a higher prevalence and diversity of blood parasites compared with partially migratory birds such as Emberiza schoeniclus. Notably, an age-dependent difference in parasite prevalence was observed in Acrocephalus spp., but not in E. schoeniclus. Nematodes were absent in all examined bird species. Proposed transmission areas were identified for nine haemosporidian lineages, showing three different types of transmission area, either with limited transmission in Europe or Africa, or active transmission in both regions.
候鸟在旅途中会遇到比本地鸟类更多的寄生虫,并将它们带到很远的地方。虽然有些寄生虫分布广泛,但传播并非不可避免,这取决于是否存在有能力的节肢动物媒介以及寄生虫与本地鸟类物种的兼容性。区分寄生虫分布区和传播区对于监测和评估本地鸟类面临的风险至关重要,因为分布区在适当的条件下可能成为潜在的传播区。本研究对在德国汉堡 "Die Reit "自然保护区采集的 455 只芦苇属鸟类的血液样本进行了筛查,目标是血孢子虫寄生虫、锥虫和丝状线虫。通过确定候鸟年龄来确定检测到的寄生虫的传播区域。根据留鸟和幼鸟提供的信息以及合格病媒的调查结果确定传播区域。与Emberiza schoeniclus等部分候鸟相比,Acrocephalus属的长途迁徙鸟类血液寄生虫的流行率和多样性更高。值得注意的是,在 Acrocephalus 属鸟类中,寄生虫的流行率与年龄有关,而在 E. schoeniclus 中则没有。所有检查过的鸟类都没有线虫。为九个血孢子虫品系确定了拟议的传播区域,显示出三种不同类型的传播区域,要么在欧洲或非洲传播有限,要么在这两个地区传播活跃。
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引用次数: 0
Evidence for a constitutive cost of host resistance on body fat growth in ewe lambs from lines selected for resistance or susceptibility to experimental infections with Haemonchus contortus. 有证据表明,宿主抗性对母羊羔羊体脂增长的构成性成本来自于对实验性传染性口蹄疫感染的抗性或易感性选育出的品系。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-26 DOI: 10.1016/j.ijpara.2024.09.004
Frédéric Douhard, Xavier Matthey, Didier Marcon, Camille Coffre-Thomain, Lucie Estivalet, Delphine Serreau, Fabrice Guégnard, Guillaume Sallé, Papa Moussa Drame, Frédéric Elleboudt, François Lecompte, Hans Adriaensen

Although benefits of selection for host resistance to gastro-intestinal nematodes have long been recognized, its costs on production traits remain unclear. A main difficulty when studying those costs is to disentangle genetic effects due to selection from plastic responses induced by infection. Putative costs of host resistance have been extensively investigated in growing sheep. However, while most of those studies have relied on live weight to assess body growth, more comprehensive assessments accounting for body composition are advocated to detect trade-offs. In this study we used 90 female lambs from lines divergently selected on resistance to Haemonchus contortus that we experimentally infected (n = 60) or not (n = 30) under controlled conditions. As those conditions were defined to enable uninfected lambs to fully express their growth potential, we sought to precisely identify the effects of selection for host resistance on health traits and on growth traits. We assessed muscular and fat growth based on repeated measurements with dorsal ultrasonography for all lambs on farm, and with whole-body computed tomography (CT) scans for a subgroup of 18 infected lambs. Lambs achieved a high growth rate, including infected ones despite their high worm burden (confirmed at necropsy in the subgroup). As expected, lambs from the resistant (R) line were less infected than those from the susceptible (S) line. However, the clear pathogenic effects observed on muscular growth and voluntary feed intake were similar between lines. In contrast, a line difference in body fat was supported both by dorsal and volumetric CT measurements. Specifically, lower fat in the R line compared with the S line was observed equally in infected and uninfected groups, thus providing evidence for a constitutive cost of host resistance. Although this cost is not necessarily disadvantageous in nutrient-rich environments exposing animals to excess fat deposition, its consequences in nutrient-scarce environments may be important to promote sustainable breeding strategies for host resistance.

尽管宿主对胃肠道线虫的抗性选择所带来的益处早已得到认可,但其对生产性状所造成的代价仍不清楚。研究这些成本的一个主要困难是如何将选择的遗传效应与感染引起的可塑性反应区分开来。在绵羊生长过程中,对宿主抗性的成本进行了广泛研究。然而,虽然这些研究大多依赖活重来评估身体生长情况,但我们主张对身体组成进行更全面的评估,以检测权衡。在这项研究中,我们使用了 90 只雌性羔羊,这些羔羊来自对血包虫病具有抗性的不同选育品系,我们在受控条件下对它们进行了感染(n = 60)或不感染(n = 30)的实验。由于这些条件是为了让未感染的羔羊充分发挥其生长潜力,因此我们试图精确确定宿主抗性选择对健康性状和生长性状的影响。我们对农场所有羔羊的肌肉和脂肪生长情况进行了评估,这些评估是通过背侧超声波重复测量进行的,并对 18 只受感染羔羊进行了全身计算机断层扫描(CT)。羔羊的生长率很高,包括受感染的羔羊,尽管它们的虫体负担很重(在子组的尸体解剖中得到证实)。不出所料,抗性(R)品系羔羊的感染率低于易感(S)品系羔羊。然而,在肌肉生长和自愿采食量方面观察到的明显致病影响在不同品系之间是相似的。与此相反,体脂的品系差异得到了背侧和容积 CT 测量的支持。具体来说,与 S 系相比,R 系的脂肪较低,这一点在感染组和未感染组中都能观察到,从而为宿主抵抗力的构成代价提供了证据。虽然在营养丰富的环境中,这种代价不一定会使动物脂肪沉积过多,但在营养匮乏的环境中,其后果可能对促进可持续的抗宿主育种策略非常重要。
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引用次数: 0
Parasite β-diversity along a stream: effect of distance and environment. 溪流寄生虫的β多样性:距离和环境的影响。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-25 DOI: 10.1016/j.ijpara.2024.09.002
Verónica Taglioretti, María Alejandra Rossin, Eugenia Levy, Juan Tomás Timi

Parasites can provide suitable models for studying β-diversity due to their strict dependence on both the environment and the biology and distribution of their hosts, aiding in the interpretation of any patterns that hosts can display. With the aim of quantifying the relative importance of host features, environmental factors and spatial distances as drivers of fish parasite β-diversity along a unidirectional gradient, the structure of fish parasite assemblages was analysed using generalised dissimilarity models (GDMs). A total of 150 poeciliid fish were examined for larval trematodes, recording host features and physical parameters of each sampling site along the stream. Differences among digenean communities increased when Strahler order changed along the stream, associated with increasing species richness and abundance downstream. Environmental gradient, spatial distance and host features were identified as significant determining factors of species turnover, with conductivity being the most important, followed by spatial distance. In the present study, environmental variables were spatially structured along the stream, their effects as structurers of parasite β-diversity being higher than the pure environmental or the pure distance effect. Such predominance prevents us from establishing at what point on the continuum from niche to neutrality these communities are located. Results from the present research contribute to improving our knowledge of the factors that shape parasite community changes, and underline the importance of considering the pure and shared effects of spatial, environmental and host feature factors in order to determine the real contribution of each one as a determinant of parasite β- diversity.

由于寄生虫严格依赖于环境及其宿主的生物学和分布,因此寄生虫可以为研究β多样性提供合适的模型,有助于解释宿主可能显示的任何模式。为了量化宿主特征、环境因素和空间距离作为单向梯度鱼类寄生虫β多样性驱动因素的相对重要性,我们使用广义相似性模型(GDMs)分析了鱼类寄生虫群的结构。共对 150 条poeciliid 鱼进行了幼虫吸虫检查,并记录了溪流沿岸每个采样点的寄主特征和物理参数。当溪流的斯特雷勒顺序发生变化时,地衣类群落之间的差异也随之增加,这与下游物种丰富度和丰度的增加有关。环境梯度、空间距离和寄主特征被认为是物种更替的重要决定因素,其中电导率最为重要,空间距离次之。在本研究中,环境变量沿溪流呈空间结构分布,它们对寄生虫 β 多样性的影响高于纯环境或纯距离的影响。这种优势使我们无法确定这些群落位于从利基到中性的连续统一体的哪一个点上。本研究的结果有助于提高我们对影响寄生虫群落变化的因素的认识,并强调了考虑空间、环境和宿主特征因素的纯效应和共同效应的重要性,以便确定每种因素作为寄生虫β多样性决定因素的真正贡献。
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引用次数: 0
The invasive acanthocephalan parasite Pachysentis canicola is associated with a declining endemic island fox population on San Miguel Island. 入侵的棘头蚴寄生虫 Pachysentis canicola 与圣米格尔岛上的特有岛狐数量下降有关。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-24 DOI: 10.1016/j.ijpara.2024.09.003
O Alejandro Aleuy, Leslie W Woods, Benjamin J Padilla, Dennis Richardson, Juliann T Schamel, Stacy Baker, Martín García-Varela, Charlotte Hammond, Sarah P Lawson, Jasmine N Childress, Jason Rohr, Kevin D Lafferty

In the late 1990s, the San Miguel Island fox (Urocyon littoralis littoralis) faced near-extinction. Fourteen of the 15 remaining foxes were placed into an island-based captive breeding program used to repopulate the island. Although the fox population in San Miguel reached pre-decline numbers by 2010, a second decline started around 2014, coincidental with a newly observed acanthocephalan parasite. To identify this introduced acanthocephalan species and determine the pathologic consequences of its infection on the health of foxes, we used an extensive record of island fox necropsies and associated parasite collections. In addition, we used detailed fox capture-recapture data to investigate population health and demographic trends of foxes before and after parasite emergence. We identify the parasite as Pachysentis canicola, a common acanthocephalan in mainland foxes in North America. The parasite was detected in 69% of the necropsied foxes from San Miguel Island and was not found in any of the other five Channel Island fox subspecies. Health impacts attributed to the acanthocephalan parasite, including erosive and ulcerative enteritis, transmural necrosis, and inflammation, were described in 47% of the foxes infected with the acanthocephalan. Despite infection with various other helminth parasite species, body condition remained good and the mortality rate low in San Miguel Island foxes until the arrival of the acanthocephalan. Body condition improved after 2018, perhaps due to increases in rainfall following a drought, but remained 27% lower than the pre-acanthocephalan period, which suggests that environmental conditions and parasitism jointly drive fox population dynamics.

20 世纪 90 年代末,圣米格尔岛狐(Urocyon littoralis littoralis)濒临灭绝。在仅存的 15 只狐狸中,有 14 只被放入岛上的圈养繁殖计划中,用于重新繁殖岛上的狐狸。虽然圣米格尔岛的狐狸数量在 2010 年达到了衰退前的水平,但在 2014 年左右又开始了第二次衰退,这与一种新观察到的棘头蚴寄生虫不谋而合。为了确定这种引入的棘头蚴物种,并确定其感染对狐狸健康造成的病理后果,我们使用了大量的岛上狐狸尸体解剖记录和相关寄生虫采集记录。此外,我们还利用详细的狐狸捕获再捕获数据,调查了寄生虫出现前后狐狸的种群健康状况和人口趋势。我们确定寄生虫为 Pachysentis canicola,这是北美大陆狐狸中常见的棘头蚴。在圣米格尔岛 69% 的尸体解剖狐狸中检测到了这种寄生虫,而在海峡岛其他五个狐狸亚种中均未发现。47%感染棘头蚴寄生虫的狐狸的健康受到了影响,包括侵蚀性和溃疡性肠炎、跨膜坏死和炎症。尽管感染了其他多种蠕虫寄生虫,但在棘头蚴到来之前,圣米格尔岛狐狸的身体状况一直很好,死亡率也很低。2018 年后,身体状况有所改善,这可能是由于干旱后降雨量增加所致,但仍比尖头蚴感染前低 27%,这表明环境条件和寄生虫共同驱动着狐狸的种群动态。
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引用次数: 0
Molecular characterisation of Australasian Ixodiphagus (Hymenoptera; Encyrtidae; Encyrtinae) reveals unexpected diversity and a potential novel host switch. 澳大拉西亚 Ixodiphagus(膜翅目;Encyrtidae;Encyrtinae)的分子特征揭示了意想不到的多样性和潜在的新型宿主转换。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-11 DOI: 10.1016/j.ijpara.2024.09.001
Madalene M Giannotta, Ina Smith, Michelle Michie, Kim Blasdell, Mike Dunn, James Nicholls, Allen C G Heath, Juanita Rodriguez, Alexander W Gofton

Ticks are important medical and veterinary parasites that represent a substantial health threat to humans, companion animals, and livestock. Ixodiphagus wasps (Hymenoptera; Encyrtidae) are known endoparasitoids of ixodid (hard) and argasid (soft) ticks, with potential utility as natural biocontrol agents. Two species, Ixodiphagus brunneus and Ixodiphagus mysorensis, are previously recorded from Australia, however, the genus lacks formal revisionary work in Australia, and the validity and host ranges of these species remain uncertain. This work aimed to investigate the diversity of Ixodiphagus in Australasia and provide a molecular data resource for future work on these understudied endoparasitoids. We extracted DNA from archival Ixodiphagus specimens from Australian and New Zealand insect collections and performed high-throughput sequencing which resulted in complete or mostly complete mitochondrial genome sequences from 11 specimens, including I. brunneus, Ixodiphagus taiaroaensis, and a novel Ixodiphagus sp. reared from Rhipicephalus linnaei from Townsville, Australia. In addition, approximately 70% of the genome of the Wolbachia endosymbiont of I. brunneus was recovered. Finally, we screened 178 recently collected pooled tick samples from southern New South Wales, Australia, for Ixodiphagus spp. using 28S rRNA and cytochrome c oxidase subunit 1(COI) gene PCR, and recovered 14 positive samples. Phylogenetic analysis of Australasian Ixodiphagus spp. based on 28S rRNA and complete mitochondrial genome sequences determined that members of the Australasian fauna are distinct from Ixodiphagus hookeri (the only other Ixodiphagus species for which genetic data exists), and that at least two distinct species are present in Australia; I. brunneus identified from Ixodes holocyclus and Haemaphysalis bancrofti ticks, and an uncharacterised Ixodiphagus sp. found in Rhipicephalus linnaei ticks from northern Queensland. Furthermore, there was substantial genetic diversity at the 28S rRNA loci among I. brunneus samples, which may represent normal genetic variability or a secondary cryptic species. The molecular data generated here represents the first known for the genus Ixodiphagus in Australasia, doubling that of the world fauna, and provides the first known complete mitochondrial genomes for these important tick parasitoids.

蜱是重要的医学和兽医寄生虫,对人类、伴侣动物和牲畜的健康构成严重威胁。Ixodiphagus 黄蜂(膜翅目;Encyrtidae)是已知的蜱(硬蜱)和蜱(软蜱)的内寄生虫,具有天然生物控制剂的潜在用途。澳大利亚以前曾记录过两个物种,即 Ixodiphagus brunneus 和 Ixodiphagus mysorensis,但该属在澳大利亚缺乏正式的修订工作,这些物种的有效性和宿主范围仍不确定。这项研究旨在调查澳大拉西亚 Ixodiphagus 的多样性,并为今后研究这些研究不足的内寄生虫提供分子数据资源。我们从澳大利亚和新西兰昆虫馆藏的 Ixodiphagus 标本中提取了 DNA,并进行了高通量测序,从而获得了 11 个标本的完整或基本完整的线粒体基因组序列,其中包括 I. brunneus、Ixodiphagus taiaroaensis,以及从澳大利亚汤斯维尔的 Rhipicephalus linnaei 上饲养的新型 Ixodiphagus sp.。此外,我们还恢复了布鲁内斯虹鳟 Wolbachia 内共生体约 70% 的基因组。最后,我们利用 28S rRNA 和细胞色素 c 氧化酶亚单位 1(COI)基因 PCR 技术对最近从澳大利亚新南威尔士州南部采集的 178 份蜱样本进行了筛选,发现了 14 份阳性样本。基于 28S rRNA 和完整线粒体基因组序列的澳大拉西亚 Ixodiphagus spp.系统进化分析表明,澳大拉西亚动物群的成员与 Ixodiphagus hookeri(唯一有遗传数据的其他 Ixodiphagus 物种)不同,澳大利亚至少有两个不同的物种:I.从 Ixodes holocyclus 和 Haemaphysalis bancrofti 的蜱虫中发现了 Ixodiphagus brunneus,从昆士兰北部的 Rhipicephalus linnaei 的蜱虫中发现了一种未定性的 Ixodiphagus sp.。此外,在 I. brunneus 样本中,28S rRNA 位点存在大量遗传多样性,这可能代表正常的遗传变异或次生隐性物种。这里产生的分子数据是已知澳大拉西亚 Ixodiphagus 属的第一个数据,是世界动物群的两倍,并为这些重要的蜱寄生虫提供了第一个已知的完整线粒体基因组。
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引用次数: 0
Challenging the phylogenetic relationships among Echinococcus multilocularis isolates from main endemic areas 挑战主要流行区多棘球蚴分离株之间的系统发育关系。
IF 3.7 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-01 DOI: 10.1016/j.ijpara.2024.05.004

Alveolar echinococcosis (AE) is a rare but severe disease that affects more than 18,000 people worldwide per year. The complete sequencing of the mitochondrial genome of Echinococcus multilocularis has made it possible to study the genetic diversity of the parasite and its spatial and temporal evolution. We amplified the whole mitochondrial genome by PCR, using one uniplex and two multiplex reactions to cover the 13,738 bp of the mitogenome, and then sequenced the amplicons with Illumina technology. In total, 113 samples from Europe, Asia, the Arctic and North America were analyzed. Three major haplogroups were found: HG1, which clustered samples from Alaska (including Saint-Lawrence Island), Yakutia (Russia) and Svalbard; HG2, with samples from Asia, North America and Europe; and HG3, subdivided into three micro-haplogroups. HG3a included samples from North America and Europe, whereas HG3b and HG3c only include samples from Europe. In France, HG3a included samples from patients more recently diagnosed in a region outside the historical endemic area. A fourth putative haplogroup, HG4, was represented by only one isolate from Olkhon Island (Russia). The increased discriminatory power of the complete sequencing of the E. multilocularis mitogenome has made it possible to highlight four distinct geographical clusters, one being divided into three micro-haplogroups in France.

肺泡棘球蚴病(AE)是一种罕见但严重的疾病,全世界每年有 18,000 多人患病。对多棘球蚴线粒体基因组的完整测序使得研究这种寄生虫的遗传多样性及其时空演变成为可能。我们通过 PCR 扩增了整个线粒体基因组,使用了一个单工反应和两个多工反应来覆盖有丝分裂基因组的 13,738 bp,然后利用 Illumina 技术对扩增子进行了测序。共分析了来自欧洲、亚洲、北极和北美洲的 113 个样本。结果发现了三个主要的单倍群:HG1 聚集了来自阿拉斯加(包括圣劳伦斯岛)、雅库特(俄罗斯)和斯瓦尔巴群岛的样本;HG2 聚集了来自亚洲、北美和欧洲的样本;HG3 则细分为三个微型单倍群。HG3a 包括北美和欧洲的样本,而 HG3b 和 HG3c 只包括欧洲的样本。在法国,HG3a 包括最近在历史流行区以外地区确诊的患者样本。第四个可能的单倍群 HG4 仅有一个来自俄罗斯奥尔洪岛的分离株。多角体细胞有丝分裂基因组的完整测序提高了判别能力,从而突出了四个不同的地理集群,其中一个在法国被分为三个微单倍群。
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引用次数: 0
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International journal for parasitology
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