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Design and evaluation of vaccines for the control of the etiological agent of East Coast fever. 设计和评估用于控制东海岸热病原体的疫苗。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-20 DOI: 10.1186/s13071-024-06517-w
José de la Fuente, Isidro Sobrino, Margarita Villar

East Coast fever is a tick-borne theileriosis caused by Theileria parva, a protozoan parasite with the primary vector being the tick Rhipicephalus appendiculatus. This disease poses significant challenges in sub-Saharan Africa, leading to severe economic losses by causing the death of over one million livestock annually. Current control measures include vector control with acaricides and the "infection and treatment" method, which involves immunization with live sporozoites of the pathogen and treatment with long acting oxytetracycline. Despite their effectiveness, these methods face scalability and usability issues, necessitating the development of new prevention strategies, particularly in the field of vaccines for the effective and sustainable control of East Coast fever. In this primer focus, East Coast fever serves as a case study to highlight recent concepts and advancements in tick and tick-borne disease vaccine research. Vaccine design and evaluation processes are reviewed, encompassing the utilization of omics datasets and knowledge on vectors and pathogens, and exploring new design methods, such as quantum vaccinomics and messenger RNA (mRNA)-based vaccines. Key limitations and areas requiring further research are addressed, including insufficient understanding of host-pathogen molecular interactions, the impact of post-translational modifications, and vaccine efficacy variability across different trials. Additionally, new research objectives are proposed to address East Coast fever but with possible impact on other tick-borne diseases. It includes advancing knowledge on tick-pathogen-host molecular interactions, studying tick microbiota, developing novel design approaches, such as combining tick and pathogen epitopes in chimeric vaccines (exemplified by the q38-p67c case), and exploring new immunological enhancers and delivery platforms.

东海岸热是一种由蜱虫引起的蜱虫病,是一种原生动物寄生虫,主要传播媒介是蜱虫 Rhipicephalus appendiculatus。这种疾病给撒哈拉以南非洲地区带来了巨大挑战,每年造成 100 多万头牲畜死亡,造成严重的经济损失。目前的控制措施包括用杀螨剂控制病媒,以及 "感染和治疗 "法,即用病原体的活孢子虫进行免疫,并用长效土霉素进行治疗。尽管这些方法很有效,但也面临着可扩展性和可用性问题,因此有必要开发新的预防策略,特别是在疫苗领域,以有效、可持续地控制东海岸热。在这本重点介绍东海岸热的入门读物中,以东海岸热为案例,重点介绍了蜱和蜱传疾病疫苗研究的最新理念和进展。回顾了疫苗的设计和评估过程,包括利用omics 数据集以及关于病媒和病原体的知识,并探索新的设计方法,如量子疫苗组学和基于信使 RNA (mRNA) 的疫苗。研究还探讨了需要进一步研究的主要局限性和领域,包括对宿主与病原体分子相互作用的认识不足、翻译后修饰的影响以及不同试验中疫苗疗效的差异。此外,还针对东海岸热提出了新的研究目标,但可能对其他蜱传疾病产生影响。其中包括增进对蜱-病原体-宿主分子相互作用的了解,研究蜱微生物群,开发新的设计方法,如在嵌合疫苗中结合蜱和病原体表位(以 q38-p67c 为例),以及探索新的免疫增强剂和递送平台。
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引用次数: 0
Resistance intensity test (RIT): a novel bioassay for quantifying the level of acaricide resistance in Rhipicephalus microplus ticks. 抗药性强度测试 (RIT):一种用于量化 Rhipicephalus microplus 蜱对杀螨剂抗药性水平的新型生物测定方法。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-20 DOI: 10.1186/s13071-024-06561-6
Frans Jongejan, Laura Berger, Laura Homminga, Iris Hulsebos, Alita Petersen, Priscila Teixeira Ferreira, José Reck, Guilherme Klafke

Background: One bioassay for detecting acaricide resistance in livestock ticks is the adult immersion test (AIT), wherein engorged ticks are briefly immersed into a solution of a particular acaricidal compound and examined for mortality, their egg-laying capacity and offspring hatchability in vitro. Usually, the recommended label dose or an established discriminating dose of an acaricide is used to determine high mortality (≥ 95%) of susceptible tick strains. Such a test intends to detect the presence of resistance in a tick population. However, the adult immersion test does not directly translate the bioassay results to the predicted efficacy in the field. In this paper, we used the AIT as an initial screening bioassay supplemented with the resistance intensity test (RIT), a novel larval-based bioassay, wherein the resistance level can be determined and translated to the expected field efficacy. This was done by adopting World Health Organisation (WHO) guidelines for resistance detection in mosquitoes, which combines a 1 × recommended dose with 5 × and 10 × concentrated doses to reveal low, moderate and high resistance intensity, respectively.

Methods: Engorged Rhipicephalus microplus ticks were collected from cattle at six different ranches across Rio Grande do Sul, Brazil, as part of the state's acaricide resistance surveillance program. Groups of adult ticks from each field collection were subjected to the AIT from each field sample. Additionally, engorged female ticks from each ranch were allowed to lay eggs, and their larval progeny aged 14 to 28 days were then used in the RIT. Deltamethrin and a combination of cypermethrin, chlorpyrifos and piperonyl butoxide were used in both tests, and the results were statistically analysed.

Results: The in vitro efficacy of deltamethrin against adult ticks in the AIT ranged between 8.74% and 25.38%. The corresponding RIT results on their larval progeny indicated a high resistance level. In the immersion test, the in vitro efficacy of the combination of cypermethrin, chlorpyrifos, and piperonyl butoxide against adult ticks ranged between 49.31% and 100%. The corresponding RIT results on their larval progeny indicated a similar response ranging from fully susceptible to low or moderate resistance. The Pearson correlation coefficient (r = 0.883) showed a high correlation between tick mortality at the 1 × recommended concentrations of acaricides in both tests.

Conclusions: The resistance intensity test is a valuable addition to the range of bioassays currently available for detecting acaricide resistance by determining the level of acaricide resistance. This is relevant to whether or not to continue using a particular acaricidal class for controlling cattle ticks.

背景:检测家畜蜱对杀螨剂抗药性的一种生物测定方法是成蜱浸泡试验(AIT),将充血的蜱短暂浸入特定杀螨化合物溶液中,在体外检测其死亡率、产卵能力和后代孵化率。通常,使用杀螨剂的推荐标签剂量或既定鉴别剂量来确定易感蜱株的高死亡率(≥ 95%)。这种试验旨在检测蜱群中是否存在抗药性。然而,成虫浸泡试验并不能直接将生物测定结果转化为预测的田间药效。在本文中,我们使用成虫浸泡试验作为初步筛选生物测定,并辅以抗药性强度试验(RIT),这是一种基于幼虫的新型生物测定,可确定抗药性水平并将其转化为预期的田间药效。为此,采用了世界卫生组织(WHO)的蚊虫抗药性检测准则,将 1 倍推荐剂量与 5 倍和 10 倍浓缩剂量相结合,分别显示低、中和高抗药性强度:方法:作为巴西南里奥格兰德州杀螨剂抗药性监测计划的一部分,从该州六个不同牧场的牛身上采集了啮齿类 Rhipicephalus microplus蜱。从每个牧场采集的成蜱组都接受了来自每个牧场样本的 AIT 检测。此外,还让每个牧场的充血雌蜱产卵,然后将其 14 至 28 天的幼虫后代用于 RIT。两项试验均使用了溴氰菊酯以及氯氰菊酯、毒死蜱和胡椒基丁醚的组合,并对结果进行了统计分析:在 AIT 中,溴氰菊酯对成蜱的体外效力在 8.74% 和 25.38% 之间。对其幼虫后代的相应 RIT 结果表明抗药性水平很高。在浸泡试验中,氯氰菊酯、毒死蜱和胡椒基丁醚对成蜱的体外药效在 49.31% 和 100% 之间。对其幼虫后代的相应 RIT 结果也显示出类似的反应,从完全易感到低度或中度抗性。皮尔逊相关系数(r = 0.883)显示,在这两项试验中,1 × 建议浓度杀螨剂的蜱死亡率之间存在高度相关性:通过确定杀螨剂的抗药性水平,抗药性强度试验是对目前可用来检测杀螨剂抗药性的一系列生物测定方法的重要补充。这关系到是否继续使用某种杀螨剂来控制牛蜱。
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引用次数: 0
Natural variation in timing of egg hatching, response to water agitation, and bidirectional selection of early and late hatching strains of the malaria mosquito Anopheles gambiae sensu lato. 疟蚊冈比亚按蚊卵孵化时间的自然变异、对水搅拌的反应以及早期和晚期孵化品系的双向选择。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-20 DOI: 10.1186/s13071-024-06533-w
Emmanuel Chinweuba Ottih, Frederic Tripet

Background: Eggs of anopheline mosquitoes hatch within a few days of laying and require high levels of humidity to survive. Assessing natural variation in egg hatching and its environmental and genetic determinants in sibling species of the malaria vector Anopheles gambiae s.l. is important for understanding their adaptation to variable aquatic habitats. Crucially, it can also inform insectary rearing practices toward the optimization of mosquito production for genetic vector control strategies.

Methods: Hatching rates and timing of egg hatching in long-established and recently colonized strains of An. gambiae s.s, Anopheles arabiensis, and Anopheles coluzzii, were compared under still water conditions (26 ℃) and with cold (4 ℃) and (15 ℃) water agitation regimes. Next, early and late hatching strains of the recently colonized An. coluzzii VK colony were generated through bidirectional selection for 18-23 generations to detect a genetic component for this trait.

Results: Hatching rates differed significantly between species and treatments. The older An. arabiensis Senn and An. gambiae s.s. Kisumu strains had the highest proportion of hatching and preferred the nonagitation treatment at 26 °C. In contrast, the more recently colonized An. coluzzii VK and An. arabiensis Rufisque strains had lower overall hatching success but responded strongly to agitation at 4 °C, while the An. coluzzii Mopti strain did not significantly respond to water agitation. In all strains, eggs hatching started at day 2 and continued till day 5 in the older strains, whilst it was more staggered and extended up to day 6 in the younger strains. Bidirectional selection for early and late hatching over many generations resulted in early hatching selected strains with eggs hatching 2-3 days earlier than in late hatching ones indicating a significant heritable component for these traits.

Conclusions: Water agitation and temperature and age of colonization are likely important determinants of egg hatching in natural An. gambiae s.l.

Populations: Current rearing protocols systematically select for fast hatching and result in the progressive loss of staggered egg hatching in older laboratory strains. The selection of novel slow-hatching strains may prove instrumental to enable the mass production, shipping, and release of Anopheles mosquitoes across Africa as part of genetic vector control programs.

背景:按蚊的卵在产卵后几天内孵化,需要高湿度才能存活。评估疟疾病媒冈比亚按蚊同胞种卵孵化的自然变异及其环境和遗传决定因素,对于了解它们对多变水生生境的适应性非常重要。方法:比较了在静水条件下(26 ℃)和在冷水(4 ℃)和(15 ℃)水搅拌条件下,长期定居和最近定居的冈比亚按蚊、阿拉伯按蚊和科鲁兹按蚊的孵化率和卵孵化时间。接下来,通过18-23代的双向选择,产生了新近定殖的Coluzzii鳗VK群的早孵化和晚孵化品系,以检测该性状的遗传成分:结果:不同物种和处理的孵化率差异很大。较老的阿拉伯疟蚊(An. arabiensis Senn)和冈比亚疟蚊(An. gambiae s.s.Kisumu)菌株的孵化率最高,并且更喜欢 26 °C的非诱导处理。相比之下,新近定殖的科鲁兹疟原虫 VK 株系和阿拉伯疟原虫 Rufisque 株系的总体孵化成功率较低,但对 4 °C下的搅拌反应强烈,而科鲁兹疟原虫莫普提株系对水的搅拌反应不明显。在所有品系中,老品系的卵孵化从第2天开始,一直持续到第5天,而年轻品系的卵孵化则更错开,一直持续到第6天。经过多代早孵和晚孵的双向选择,早孵品系的孵卵时间比晚孵品系早2-3天,这表明这些性状有显著的遗传因素:结论:水的搅拌、温度和定殖年龄可能是冈比亚安虫自然种群卵孵化的重要决定因素:目前的饲养方案系统性地选择快速孵化,导致老的实验室品系逐渐丧失交错卵孵化。选择新的慢孵化品系可能有助于在非洲大规模生产、运输和释放按蚊,作为遗传病媒控制项目的一部分。
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引用次数: 0
Diversity and prevalence of Leucocytozoon in black flies (Diptera: Simuliidae) of Thailand. 泰国黑蝇(双翅目:蚋科)中白细胞介素的多样性和流行率。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-19 DOI: 10.1186/s13071-024-06567-0
Waraporn Jumpato, Wannachai Wannasingha, Chavanut Jaroenchaiwattanachote, Ronnalit Mintara, Komgrit Wongpakam, Peter H Adler, Pairot Pramual
<p><strong>Background: </strong>Leucocytozoonosis, a parasitic disease of birds, is caused by haemosporidian protozoan parasites of the genus Leucocytozoon, which infect diverse avian species, including poultry. These parasites are transmitted by several black fly species, but knowledge of the factors determining the diversity and prevalence in these vectors, which is crucial for fully understanding disease epidemiology, is largely unexplored. In this study, we investigated factors associated with the prevalence and diversity of Leucocytozoon species in black flies from Thailand.</p><p><strong>Methods: </strong>Adults of two black fly taxa (Simulium asakoae Takaoka and Davies complex and S. khelangense Takaoka, Srisuka and Saeung) were collected using sweep nets at nine locations in northern and northeastern regions of Thailand. Specimens were identified morphologically and the results corroborated by DNA barcoding. Molecular methods using specific primers for amplification of the mitochondrial cytochrome b (cyt b) gene of Leucocytozoon were used to detect the parasite in black flies. Species and lineages of Leucocytozoon were determined using the MalAvi database of malaria parasites and related haemosporidians in avian hosts. Regression analysis was used to examine relationships between Leucocytozoon diversity and prevalence, black fly abundance and habitat characteristics.</p><p><strong>Results: </strong>A total of 11,718 adult black flies were collected, of which 4367 were members of the S. asakoae complex and 7351 were S. khelangense. For molecular detection of Leucocytozoon, we randomly selected 300 individual female black flies of the S. asakoae complex and 850 females of S. khelangense pooled into groups of five individuals (= 170 pools). A total of 34 of the 300 specimens of the S. asakoae complex and 118 of the 170 pools of S. khelangense were positive for Leucocytozoon. Fifty-four lineages (haplotypes) were identified, all of which belonged to those reported in domestic chickens, Gallus gallus, with one exception that was identified in S. khelangense and found to be closely related to the Leucocytozoon lineages reported in owls; this is the first record of the latter lineage in Asian black flies. Among these haplotypes, nine and 45 were exclusively found in the S. asakoae complex and S. khelangense, respectively. No lineage was shared between these black fly taxa. Analysis of similarity (ANOSIM) revealed significant Leucocytozoon lineage composition between the two black flies. Phylogenetic analysis found that Leucocytozoon lineages in the S. asakoae complex and S. khelangense are largely isolated, agreeing with the ANOSIM result. The overall prevalence of Leucocytozoon in the S. asakoae complex was 11.3% and ranged from 9% to 13% in each collection. Leucocytozoon prevalence in S. khelangense was 21%, varying from 13% to 37% in each collection. The Shannon H' index indicated greater Leucocytozoon diversity in S. khelangense (H' = 3.044)
背景:白细胞虫病是一种鸟类寄生虫病,由白细胞虫属血孢子原生动物寄生虫引起,感染包括家禽在内的多种禽类。这些寄生虫由几种黑蝇传播,但决定这些病媒多样性和流行率的因素对充分了解疾病流行病学至关重要,但这方面的知识在很大程度上尚未得到探索。在这项研究中,我们调查了与泰国黑蝇中白细胞虫种类的流行和多样性有关的因素:方法:在泰国北部和东北部地区的 9 个地点使用扫网收集了两个黑蝇类群(Simulium asakoae Takaoka and Davies complex 和 S. khelangense Takaoka, Srisuka and Saeung)的成虫。对标本进行了形态鉴定,并通过 DNA 条形码对鉴定结果进行了确认。使用特异引物扩增线粒体细胞色素 b(cyt b)基因的分子方法检测了黑蝇中的寄生虫。利用 MalAvi 数据库确定了禽类宿主中的疟原虫和相关血孢子虫的种类和血系。采用回归分析法研究了Leucocytozoon多样性和流行率、黑蝇丰度和栖息地特征之间的关系:结果:共收集到 11718 只成年黑蝇,其中 4367 只为 S. asakoae 复合体成员,7351 只为 S. khelangense。为了进行白细胞介素的分子检测,我们随机选取了300只浅胁黑蝇复合体的雌性黑蝇和850只谢朗氏黑蝇的雌性黑蝇,每5只为一组(=170组)。在 300 个 S. asakoae 复合体标本和 170 个 S. khelangense 池中,共有 34 个标本和 118 个池子的白细胞介素阳性。鉴定出 54 个品系(单倍型),所有这些品系都属于在家鸡(Gallus gallus)中报告的品系,只有一个品系在 S. khelangense 中被鉴定出,并被发现与在猫头鹰中报告的 Leucocytozoon 品系密切相关;这是亚洲黑蝇中后一个品系的首次记录。在这些单倍型中,分别有 9 个和 45 个单倍型仅见于 S. asakoae 复合物和 S. khelangense。这些黑蝇类群之间没有共享的单系。相似性分析(ANOSIM)显示,这两种黑蝇之间有明显的亮孢子虫世系组成。系统进化分析发现,S. asakoae 复合体和 S. khelangense 中的 Leucocytozoon 系在很大程度上是孤立的,这与 ANOSIM 的结果一致。在 S. asakoae 复合菌群中,Leucocytozoon 的总体流行率为 11.3%,在每个菌群中的流行率从 9% 到 13% 不等。在 S. khelangense 中,白色念珠菌的流行率为 21%,每个采集物中的流行率从 13% 到 37% 不等。香农 H'指数表明,S. khelangense(H' = 3.044)中的白细胞虫多样性高于 S. asakoae 复合体(H' = 1.920)。回归分析表明,Leucocytozoon 多样性与黑蝇数量呈正相关,与最高气温呈负相关:本研究结果表明,泰国的 S. asakoae 复合体和 S. khelangense 中的 Leucocytozoon 系的流行率和多样性与这些黑蝇的数量和气温有关。鉴定出的亮孢子虫系还显示出一定程度的黑蝇类群特异性,这可能与这些媒介的不同丰度峰值有关。有利于黑蝇发展的环境条件可能是白细胞虫流行、多样性和病媒-寄生虫共同进化的驱动因素。
{"title":"Diversity and prevalence of Leucocytozoon in black flies (Diptera: Simuliidae) of Thailand.","authors":"Waraporn Jumpato, Wannachai Wannasingha, Chavanut Jaroenchaiwattanachote, Ronnalit Mintara, Komgrit Wongpakam, Peter H Adler, Pairot Pramual","doi":"10.1186/s13071-024-06567-0","DOIUrl":"https://doi.org/10.1186/s13071-024-06567-0","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Leucocytozoonosis, a parasitic disease of birds, is caused by haemosporidian protozoan parasites of the genus Leucocytozoon, which infect diverse avian species, including poultry. These parasites are transmitted by several black fly species, but knowledge of the factors determining the diversity and prevalence in these vectors, which is crucial for fully understanding disease epidemiology, is largely unexplored. In this study, we investigated factors associated with the prevalence and diversity of Leucocytozoon species in black flies from Thailand.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;Adults of two black fly taxa (Simulium asakoae Takaoka and Davies complex and S. khelangense Takaoka, Srisuka and Saeung) were collected using sweep nets at nine locations in northern and northeastern regions of Thailand. Specimens were identified morphologically and the results corroborated by DNA barcoding. Molecular methods using specific primers for amplification of the mitochondrial cytochrome b (cyt b) gene of Leucocytozoon were used to detect the parasite in black flies. Species and lineages of Leucocytozoon were determined using the MalAvi database of malaria parasites and related haemosporidians in avian hosts. Regression analysis was used to examine relationships between Leucocytozoon diversity and prevalence, black fly abundance and habitat characteristics.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;A total of 11,718 adult black flies were collected, of which 4367 were members of the S. asakoae complex and 7351 were S. khelangense. For molecular detection of Leucocytozoon, we randomly selected 300 individual female black flies of the S. asakoae complex and 850 females of S. khelangense pooled into groups of five individuals (= 170 pools). A total of 34 of the 300 specimens of the S. asakoae complex and 118 of the 170 pools of S. khelangense were positive for Leucocytozoon. Fifty-four lineages (haplotypes) were identified, all of which belonged to those reported in domestic chickens, Gallus gallus, with one exception that was identified in S. khelangense and found to be closely related to the Leucocytozoon lineages reported in owls; this is the first record of the latter lineage in Asian black flies. Among these haplotypes, nine and 45 were exclusively found in the S. asakoae complex and S. khelangense, respectively. No lineage was shared between these black fly taxa. Analysis of similarity (ANOSIM) revealed significant Leucocytozoon lineage composition between the two black flies. Phylogenetic analysis found that Leucocytozoon lineages in the S. asakoae complex and S. khelangense are largely isolated, agreeing with the ANOSIM result. The overall prevalence of Leucocytozoon in the S. asakoae complex was 11.3% and ranged from 9% to 13% in each collection. Leucocytozoon prevalence in S. khelangense was 21%, varying from 13% to 37% in each collection. The Shannon H' index indicated greater Leucocytozoon diversity in S. khelangense (H' = 3.044)","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"17 1","pages":"475"},"PeriodicalIF":3.0,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142676423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Co-expression gene module analysis in response to attenuated cercaria vaccine reveals a critical role for NK cells in protection against Schistosoma mansoni. 对减毒恙虫疫苗反应的共表达基因模块分析揭示了 NK 细胞在保护人体免受曼氏血吸虫感染中的关键作用。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-19 DOI: 10.1186/s13071-024-06505-0
Almiro Pires da Silva Neto, Juliana Vitoriano-Souza, Mariana Ivo Khouri, Regiane Degan Favaro, Robert Alan Wilson, Luciana Cezar de Cerqueira Leite, Pablo Ivan Pereira Ramos, Leonardo Paiva Farias

Background: Despite decades of research, an effective schistosomiasis vaccine remains elusive. The radiation-attenuated (RA) cercarial vaccine remains the best model for eliciting high levels of protection. We have recently explored this model in mice to identify potentially protective pathways by examining gene expression patterns in peripheral blood mononuclear cells (PBMC).

Methods: Herein, we reanalyzed the transcriptomic data from PBMC obtained from vaccinated and infected C57BL/6 mice in three timepoints (Days 7 and 17 after infection or vaccination and Day 7 post-challenge). In addition, we generated new data on PBMC collected 35 days after infection. Deconvolution analysis was performed to estimate immune cell composition by CIBERSORTx. Gene co-expression networks and over-representation analysis (ORA) were performed using the CEMiTool package. Protein-protein interaction networks were constructed using STRING, and the hub proteins for each module were identified using Cytoscape.

Results: Co-expression network analysis identified a module (M2) associated with the infection process, grouping genes related to a Th2 immune response, and a second module (M6) associated with the vaccination process, displaying pathways related to a Th1 response, CD8 + T cells and NK cells. Within each module, five hub proteins were identified based on protein-protein interaction networks. The M2 infection module revealed Chil3, Il4, Cx3cr1, Emr1 and Ccl2 as hubs, while module M6, associated with vaccination, disclosed Prf1, Klrc1, IFN-γ, Ncr1 and Tbx21 as hub proteins.

Conclusions: Our data point to the potentiald role of NK cells that may contribute to the RA vaccine response through the production of IFN-γ orchestrated by the T-bet transcription factor (Tbx21).

背景:尽管进行了数十年的研究,但有效的血吸虫病疫苗仍遥遥无期。辐射减毒(RA)carial 疫苗仍然是激发高水平保护的最佳模型。方法:在此,我们重新分析了接种疫苗和感染 C57BL/6 小鼠的 PBMC 在三个时间点(感染或接种疫苗后第 7 天和第 17 天以及挑战后第 7 天)的转录组数据。此外,我们还生成了感染后 35 天采集的 PBMC 的新数据。通过 CIBERSORTx 进行解卷积分析以估计免疫细胞的组成。使用 CEMiTool 软件包进行了基因共表达网络和过度表现分析(ORA)。使用STRING构建了蛋白质-蛋白质相互作用网络,并使用Cytoscape确定了每个模块的中心蛋白:共表达网络分析确定了一个与感染过程相关的模块(M2),该模块将与Th2免疫应答相关的基因分组;第二个模块(M6)与疫苗接种过程相关,显示了与Th1应答、CD8 + T细胞和NK细胞相关的通路。在每个模块中,根据蛋白质-蛋白质相互作用网络确定了五个枢纽蛋白。M2感染模块显示Chil3、Il4、Cx3cr1、Emr1和Ccl2为中心蛋白,而与疫苗接种相关的M6模块则显示Prf1、Klrc1、IFN-γ、Ncr1和Tbx21为中心蛋白:我们的数据表明,NK 细胞可能通过在 T-bet 转录因子(Tbx21)的协调下产生 IFN-γ 来促进 RA 疫苗反应。
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引用次数: 0
Ascorbate peroxidase modulation confirms key role in Leishmania infantum oxidative defence. 抗坏血酸过氧化物酶的调节作用证实了它在婴儿利什曼病氧化防御中的关键作用。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-18 DOI: 10.1186/s13071-024-06562-5
Isabella Fernandes Martins Santos, Douglas de Souza Moreira, Karla Ferreira Costa, Juliana Martins Ribeiro, Silvane Maria Fonseca Murta, Ana Maria Murta Santi

Background: Ascorbate peroxidase (APX) has emerged as a promising target for chemotherapy because of its absence in humans and crucial role in the antioxidant defence of trypanosomatids. APXs, which are class I haeme-containing enzymes, reduces hydrogen peroxide using ascorbate to produce water and monodehydroascorbate, thereby preventing cell damage caused by H2O2.

Methods: We aimed to create an APX-knockout L. infantum line using CRISPR/Cas9. Despite unsuccessful attempts at full knockouts, we achieved deletion of chromosomal copies post-APX episomal insertion, yielding LiΔchrAPX::LbAPX parasites. We performed phenotypic characterization to assess the impact of these genetic modifications, which included the determination of APX transcript expression levels using quantitative PCR, drug sensitivity, infectivity, and parasite survival in macrophages.

Results: Quantitative polymerase chain reaction (PCR) analysis revealed a 10- to 13-fold reduction in APX transcript expression in LiΔchrAPX::LbAPX compared with wild-type (LiWT) and APX-overexpressing (Li::Cas9::LbAPX) parasites, respectively. The episomes in those knockdown parasites remained stable even after 20 drug-free passages in vitro. Li::Cas9::LbAPX parasites showed increased resistance to trivalent antimony (SbIII) and isoniazid, reduced tolerance to H2O2, and unchanged macrophage infectivity compared with LiWT. In contrast, LiΔchrAPX::LbAPX parasites were more sensitive to SbIII and isoniazid, exhibited greater susceptibility to H2O2-induced oxidative stress, and 72 h post-infection, showed fewer infected macrophages and intracellular amastigotes compared with LiWT parasites.

Conclusions: Our findings hint at the indispensability of APX in L. infantum and raise the possibility of its potential as a therapeutic target for leishmaniasis.

背景:由于抗坏血酸过氧化物酶(APX)在人类中的缺失以及在锥虫抗氧化防御中的关键作用,APX已成为有希望的化疗靶点。APX 是 I 类含血酶,利用抗坏血酸还原过氧化氢,生成水和单脱氢抗坏血酸,从而防止 H2O2 对细胞造成损伤:我们的目标是利用CRISPR/Cas9技术创建一个APX基因敲除的幼虫品系。尽管完全敲除的尝试并不成功,但我们还是在APX外显子插入后实现了染色体拷贝的缺失,产生了LiΔchrAPX::LbAPX寄生虫。我们进行了表型鉴定,以评估这些基因修饰的影响,包括利用定量聚合酶链反应测定 APX 转录本的表达水平、药物敏感性、感染性和寄生虫在巨噬细胞中的存活率:定量聚合酶链反应(PCR)分析表明,与野生型(LiWT)寄生虫和APX过表达型(Li::Cas9::LbAPX)寄生虫相比,LiΔchrAPX::LbAPX的APX转录本表达量分别降低了10至13倍。这些基因敲除寄生虫的外显子即使在体外无药传代 20 次后仍保持稳定。与 LiWT 相比,Li::Cas9::LbAPX 寄生虫对三价锑(SbIII)和异烟肼的抗性增强,对 H2O2 的耐受性降低,巨噬细胞感染性不变。相比之下,LiΔchrAPX::LbAPX寄生虫对SbIII和异烟肼更敏感,对H2O2诱导的氧化应激表现出更大的易感性,与LiWT寄生虫相比,感染后72小时显示出更少的受感染巨噬细胞和细胞内异形体:我们的研究结果提示了 APX 在婴儿利什曼病中的不可或缺性,并提出了将其作为利什曼病治疗靶点的可能性。
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引用次数: 0
Novel insights into antioxidant status, gene expression, and immunohistochemistry in an animal model infected with camel-derived Trypanosoma evansi and Theileria annulata. 对感染了骆驼源性伊万斯锥虫和环纹丝虫的动物模型的抗氧化状态、基因表达和免疫组织化学的新认识。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-18 DOI: 10.1186/s13071-024-06564-3
Reem M Ramadan, Alaa F Bakr, Esraa Fouad, Faten F Mohammed, Azza M Abdel-Wahab, Sahar Z Abdel-Maogood, Mohamed M El-Bahy, Mai A Salem

Background: Hemoprotozoan diseases, especially trypanosomosis and theileriosis, adversely affect the productivity, growth, and performance of camels. Regular sampling and investigation of camels are challenging due to several factors. Consequently, there is a lack of knowledge on camel parasite genotyping, cytokine production, and oxidative stress parameters during infection.

Methods: The present study investigated two critical blood protozoa infecting camels in Egypt, Trypanosoma evansi and Theileria annulata, using molecular methods, specifically 18S rRNA gene analysis. Following molecular confirmation, experimental infections were induced in Swiss albino mice to assess the expression of immune response genes and oxidative stress parameters. The study further explored the correlation between histopathological alterations and inflammatory reactions in the kidney, spleen, and liver of infected mice, alongside the immunohistochemical expression of caspase-3, proliferating cell nuclear antigen (PCNA), and tumor necrosis factor (TNF).

Results: Trypanosoma evansi and T. annulata isolated from naturally infected camels were molecularly identified and deposited in GenBank under accession numbers OR116429 and OR103130, respectively. Infection with T. evansi and T. annulata caused significant adverse effects on the immune condition of infected mice, increasing the pathogenicity of the infection. This was evidenced by a significant increase in oxidative stress parameter levels in both naturally infected camels and experimentally infected mice compared to healthy controls. Furthermore, the expression of immune response genes was significantly elevated in infected mice. Immunohistochemistry analysis showed a pronounced upregulation of caspase-3, PCNA, and TNF in the infected groups relative to the control group. These findings are the first to be reported in Egypt.

Conclusions: This study successfully identified and genotyped two economically important blood protozoa, T. evansi and T. annulata, from camels in Egypt. Additionally, the experimental animal model provided valuable insights into the immune response, oxidative stress, and histopathological changes induced by these parasites, demonstrating comparable results to naturally infected camels. These findings highlight the potential of this model to study parasite-host interactions and immune responses, contributing to a better understanding of the pathogenic mechanisms of T. evansi and T. annulata infections. This model may be useful for future studies focused on disease control and therapeutic interventions.

背景:血吸虫疾病,尤其是锥虫病和丝虫病,对骆驼的生产力、生长和性能产生了不利影响。由于多种因素,对骆驼进行定期采样和调查具有挑战性。因此,人们对骆驼寄生虫的基因分型、细胞因子的产生以及感染期间的氧化应激参数缺乏了解:本研究采用分子方法,特别是 18S rRNA 基因分析,调查了埃及骆驼感染的两种重要血液原生动物--埃文斯锥虫(Trypanosoma evansi)和年轮虫(Theileria annulata)。经分子确认后,诱导瑞士白化小鼠进行实验感染,以评估免疫反应基因的表达和氧化应激参数。该研究进一步探讨了受感染小鼠肾脏、脾脏和肝脏组织病理学改变与炎症反应之间的相关性,以及 Caspase-3、增殖细胞核抗原(PCNA)和肿瘤坏死因子(TNF)的免疫组化表达:结果:从自然感染的骆驼身上分离出的Trypanosoma evansi和T. annulata经分子鉴定后存入GenBank,登录号分别为OR116429和OR103130。T.evansi和T.annulata会对受感染小鼠的免疫状况产生显著的不利影响,增加感染的致病性。与健康对照组相比,自然感染的骆驼和实验感染的小鼠的氧化应激参数水平都显著增加,这就证明了这一点。此外,受感染小鼠的免疫反应基因表达也明显升高。免疫组化分析表明,与对照组相比,感染组的 Caspase-3、PCNA 和 TNF 明显上调。这些发现在埃及尚属首次报道:本研究成功地从埃及骆驼身上鉴定出了两种具有重要经济价值的血液原生动物--T. evansi 和 T. annulata,并对其进行了基因分型。此外,实验动物模型为了解这些寄生虫引起的免疫反应、氧化应激和组织病理学变化提供了宝贵的信息,其结果与自然感染的骆驼相当。这些发现凸显了这一模型在研究寄生虫-宿主相互作用和免疫反应方面的潜力,有助于更好地了解 T. evansi 和 T. annulata 感染的致病机制。该模型可能有助于未来以疾病控制和治疗干预为重点的研究。
{"title":"Novel insights into antioxidant status, gene expression, and immunohistochemistry in an animal model infected with camel-derived Trypanosoma evansi and Theileria annulata.","authors":"Reem M Ramadan, Alaa F Bakr, Esraa Fouad, Faten F Mohammed, Azza M Abdel-Wahab, Sahar Z Abdel-Maogood, Mohamed M El-Bahy, Mai A Salem","doi":"10.1186/s13071-024-06564-3","DOIUrl":"10.1186/s13071-024-06564-3","url":null,"abstract":"<p><strong>Background: </strong>Hemoprotozoan diseases, especially trypanosomosis and theileriosis, adversely affect the productivity, growth, and performance of camels. Regular sampling and investigation of camels are challenging due to several factors. Consequently, there is a lack of knowledge on camel parasite genotyping, cytokine production, and oxidative stress parameters during infection.</p><p><strong>Methods: </strong>The present study investigated two critical blood protozoa infecting camels in Egypt, Trypanosoma evansi and Theileria annulata, using molecular methods, specifically 18S rRNA gene analysis. Following molecular confirmation, experimental infections were induced in Swiss albino mice to assess the expression of immune response genes and oxidative stress parameters. The study further explored the correlation between histopathological alterations and inflammatory reactions in the kidney, spleen, and liver of infected mice, alongside the immunohistochemical expression of caspase-3, proliferating cell nuclear antigen (PCNA), and tumor necrosis factor (TNF).</p><p><strong>Results: </strong>Trypanosoma evansi and T. annulata isolated from naturally infected camels were molecularly identified and deposited in GenBank under accession numbers OR116429 and OR103130, respectively. Infection with T. evansi and T. annulata caused significant adverse effects on the immune condition of infected mice, increasing the pathogenicity of the infection. This was evidenced by a significant increase in oxidative stress parameter levels in both naturally infected camels and experimentally infected mice compared to healthy controls. Furthermore, the expression of immune response genes was significantly elevated in infected mice. Immunohistochemistry analysis showed a pronounced upregulation of caspase-3, PCNA, and TNF in the infected groups relative to the control group. These findings are the first to be reported in Egypt.</p><p><strong>Conclusions: </strong>This study successfully identified and genotyped two economically important blood protozoa, T. evansi and T. annulata, from camels in Egypt. Additionally, the experimental animal model provided valuable insights into the immune response, oxidative stress, and histopathological changes induced by these parasites, demonstrating comparable results to naturally infected camels. These findings highlight the potential of this model to study parasite-host interactions and immune responses, contributing to a better understanding of the pathogenic mechanisms of T. evansi and T. annulata infections. This model may be useful for future studies focused on disease control and therapeutic interventions.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"17 1","pages":"474"},"PeriodicalIF":3.0,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11575088/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142668605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Endoparasites of peritoneal organs and skeletal muscles of the European wildcat (Felis silvestris) in Germany. 德国欧洲野猫(Felis silvestris)腹膜器官和骨骼肌的内寄生虫。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-18 DOI: 10.1186/s13071-024-06571-4
Katrin Bisterfeld, Marie-Kristin Raulf, Patrick Waindok, Andrea Springer, Johannes Lang, Michael Lierz, Ursula Siebert, Christina Strube

Background: For several decades, the European wildcat (Felis silvestris) has gradually been returning to the forests of Germany, mainly in the central and southwestern regions. To increase the knowledge about this threatened species, the endoparasite status of dead found specimens from Germany was surveyed.

Methods: A total of 118 wildcats were examined for endoparasites in peritoneal organs and skeletal muscles. Owing to decomposition or incomplete carcasses, 104 gastrointestinal tracts (stomachs and intestines), 101 livers with gallbladders, 99 urinary bladders, as well as kidneys of 95 and skeletal muscles of 112 specimens were available for examination. All detected parasites were identified morphologically to genus or species level, followed by molecular examinations of one to ten specimens of each parasite species.

Results: Overall endoparasite prevalence in peritoneal organs was 99.0% (103/104). Among the 99.0% (103/104) infected gastrointestinal tracts, the most frequent species were Toxocara cati (95.2% [99/104]), Hydatigera kamiyai (84.6% [88/104]), Mesocestoides litteratus (69.2% [72/104]), Strongyloides spp. (58.7% [61/104]), Cylicospirura petrowi (37.5% [39/104]), Ancylostoma tubaeforme (31.7% [33/104]), Capillaria putorii (24.0% [25/104]), and Echinococcus multilocularis (18.3% [19/104]). In 77.8% (77/99) of the urinary bladders, Capillaria plica and/or Capillaria feliscati were detected. Moreover, the liver fluke Metorchis bilis occurred in 2.0% (2/101) of the livers, and roundworm larvae (presumably Toxocara spp.) were detected in 33.0% (37/112) of the muscle samples.

Conclusions: These results show a broad spectrum of endoparasite species infecting European wildcats in Germany. It might be assumed that some of the endoparasites could pose a risk to domestic cats (Felis catus) and humans through spillover events, or may be transmitted from domestic cats to the free-ranging population, posing a potential risk to wildcats.

背景:几十年来,欧洲野猫(Felis silvestris)逐渐返回德国森林,主要集中在中部和西南部地区。为了增加对这一濒危物种的了解,我们对在德国发现的死亡标本的内寄生虫状况进行了调查:方法:共对 118 只野猫的腹腔器官和骨骼肌肉中的内寄生虫进行了检查。由于尸体腐烂或不完整,有 104 只野猫的胃肠道(胃和肠)、101 只野猫的肝脏和胆囊、99 只野猫的尿囊、95 只野猫的肾脏和 112 只野猫的骨骼肌可供检查。对所有检测到的寄生虫进行了属或种的形态鉴定,然后对每种寄生虫的一至十个标本进行了分子检测:结果:腹膜器官中寄生虫的总体流行率为 99.0%(103/104)。在 99.0%(103/104)受感染的胃肠道中,最常见的寄生虫种类为:Toxocara cati(95.2% [99/104])、Hydatigera kamiyai(84.6% [88/104])、Mesocestoides litteratus(69.2%[72/104])、Strongyloides spp.(58.7%[61/104])、Cylicospirura petrowi(37.5%[39/104])、Ancylostoma tubaeforme(31.7%[33/104])、Capillaria putorii(24.0%[25/104])和 Echinococcus multilocularis(18.3%[19/104])。在 77.8%(77/99)的膀胱中,检测到了毛囊吸虫(Capillaria plica)和/或毛囊吸虫(Capillaria feliscati)。此外,2.0%(2/101)的肝脏样本中发现了肝吸虫,33.0%(37/112)的肌肉样本中发现了蛔虫幼虫(可能是弓形虫属):这些结果表明,德国欧洲野猫感染的内寄生虫种类繁多。可以认为,其中一些内寄生虫可能会通过外溢事件对家猫(Felis catus)和人类造成危害,也可能会从家猫传播到野猫种群中,从而对野猫造成潜在威胁。
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引用次数: 0
Improving onchocerciasis elimination surveillance: trials of odour baited Esperanza Window Traps to collect black fly vectors and real-time qPCR detection of Onchocerca volvulus in black fly pools. 改进消除盘尾丝虫病的监测工作:试验用气味诱饵埃斯佩兰萨窗口诱捕器收集黑蝇载体,并对黑蝇池中的卷尾丝虫进行实时 qPCR 检测。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-18 DOI: 10.1186/s13071-024-06554-5
Monsuru A Adeleke, Kenneth N Opara, Hayward B Mafuyai, Bertram Ekejiuba Bright Nwoke, Olabanji A Surakat, Sunday B Akinde, Murphy Nwoke, Friday M Chikezie, Clement A Yaro, Ugagu Mmaduabuchi, Michael Igbe, Emeka Makata, Fatai Oyediran, Chukwuma Anyaike, Joseph Tongjura, Frances Hawkes, Zahra O Iwalewa

Background: Entomological data for onchocerciasis surveillance relies on sampling black flies through human landing collectors in the field and laboratory testing of the flies for infection using pooled screening O-150 PCR-ELISA assay. Both techniques require improvements. This study aimed to optimize the Esperanza Window Trap (EWT) for black fly collection. We tested alternative carbon dioxide (CO2) mimics to attract black flies to the traps. Additionally, we evaluated new quantitative PCR (qPCR) methods that target mitochondrial DNA markers and have been proposed to enhance the sensitivity and specificity for detecting Onchocerca volvulus infections in blackflies.

Methods: Traps baited with low, medium and high release rates of either 2-butanone or cyclopentanone as CO2 mimics were field tested against traps baited with organically generated CO2 in four ecological zones in Nigeria: Guinea savannah, derived savannah, rainforest and montane forest. The performance of EWTs baited with CO2 or in combination with 2-butanone (low release) were subsequently evaluated against the human landing collection (HLC). Trap scaling was also pilot tested by comparing two EWTs to a single HLC team. Collected black flies were used to test detection of O. volvulus in black flies using Ov ND5 real-time PCR (qPCR) compared to the conventional pool screening O-150 PCR.

Results: EWTs baited with 2-butanone caught similar numbers of black flies (Simulium damnosum s.l.) to those baited with CO2, while cyclopentanone collected significantly fewer flies in all locations. The low release of 2-butanone was the most effective overall, although HLCs collected higher numbers of black flies than EWT baited with CO2 either singly or in combination with low-release 2-butanone. The combination of two EWTs baited with CO2 and deployed 100 m apart from each other collected similar numbers of flies as one HLC. More black fly pools were positive for O. volvulus by Ov ND5 qPCR compared with O-150 PCR in derived savannah (31.15 vs. 15.57%), montane forest (11.54 vs. 0%) and rainforest (23.08 vs. 2.56%), with only one positive pool in Guinea savannah detected by both methods.

Conclusions: The 2-butanone has potential to be used in xenomonitoring as a standardized replacement for organically generated CO2. Ov ND5 qPCR detected more positive pools than O-150 PCR. The positive pools found in foci hitherto considered to have interrupted/eliminated onchocerciasis highlight the need for more sensitive and specific methods that support programmatic assessments to identify and combat recrudescence.

背景:用于监测盘尾丝虫病的昆虫学数据依赖于在野外通过人体着陆采集器对黑蝇进行采样,以及使用集中筛选 O-150 PCR-ELISA 检测法对黑蝇进行感染实验室检测。这两种技术都需要改进。本研究旨在优化用于收集黑蝇的埃斯佩兰萨窗口诱捕器(EWT)。我们测试了其他二氧化碳(CO2)模拟物,以吸引黑蝇进入诱捕器。此外,我们还评估了新的定量 PCR(qPCR)方法,该方法以线粒体 DNA 标记为目标,被建议用于提高检测黑蝇中伏蚊子感染的灵敏度和特异性:在尼日利亚的四个生态区中,使用低、中、高释放率的 2-丁酮或环戊酮作为二氧化碳模拟物的诱捕器与使用有机产生的二氧化碳作为诱饵的诱捕器进行了实地测试:在尼日利亚的四个生态区:几内亚热带草原、衍生热带草原、热带雨林和山地森林,与以有机产生的二氧化碳为诱饵的诱捕器进行了实地测试。随后,针对人类着陆采集物(HLC)评估了以二氧化碳或结合 2-丁酮(低释放量)为诱饵的 EWT 的性能。通过比较两个 EWT 和一个 HLC 小组,还对诱捕规模进行了试点测试。使用 Ov ND5 实时 PCR (qPCR)与传统的池筛 O-150 PCR 相比,对收集的黑蝇进行了检测:结果:使用 2-butanone 诱饵的电子捕捉器捕捉到的黑蝇(Simulium damnosum s.l.)数量与使用二氧化碳诱饵的电子捕捉器相似,而环戊酮在所有地点捕捉到的黑蝇数量明显较少。低释放量的 2-丁酮总体上是最有效的,尽管 HLC 比单独使用二氧化碳或与低释放量的 2-丁酮结合使用的 EWT 捕获的黑蝇数量更高。两只以二氧化碳为诱饵的 EWT 相距 100 米,其捕获的苍蝇数量与一只 HLC 相近。在热带稀树草原(31.15% 对 15.57%)、山地森林(11.54% 对 0%)和热带雨林(23.08% 对 2.56%)中,用 Ov ND5 qPCR 与 O-150 PCR 相比,有更多的黑蝇池对 O. volvulus 呈阳性反应,而在几内亚热带稀树草原,两种方法都只检测到一个阳性池:结论:2-丁酮作为有机产生的二氧化碳的标准化替代品,有望用于异种监测。Ov ND5 qPCR 比 O-150 PCR 检测到更多的阳性库。在迄今为止被认为已中断/消除盘尾丝虫病的病灶中发现的阳性库突出表明,有必要采用更灵敏、更特异的方法来支持方案评估,以确定和打击盘尾丝虫病的复发。
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引用次数: 0
Contrasting pathogen prevalence between tick and dog populations at Chornobyl. 切尔诺贝利蜱群和狗群病原体流行率的对比。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-11-17 DOI: 10.1186/s13071-024-06563-4
Megan N Dillon, Barbara A Qurollo, Rachael Thomas, Madeline E Warren, Timothy A Mousseau, Jennifer A Betz, Norman J Kleiman, Matthew Breen

Background: The 1986 disaster at the Chornobyl Nuclear Power Plant released massive amounts of radioactive material into the local environment. In addition to radiation, remediation efforts and abandonment of military-industrial complexes contributed to contamination with heavy metals, organics, pesticides and other toxic chemicals. Numerous studies have evaluated the effects of this contamination on the local ecology. However, few studies have reported the effect of this contamination on vector-borne pathogens and their hosts. In this manuscript, we characterize tick-borne pathogen presence at two sample locations within the Chornobyl Exclusion Zone, one at the Nuclear Power Plant (NPP) and another 16 km away in Chornobyl City (CC).

Methods: Ticks and whole-blood samples were collected from free-breeding dogs captured at the NPP and CC. Endpoint PCR and quantitative PCR were used to identify tick species and to assess the presence of specific tick-borne pathogens, including Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Babesia spp., Bartonella spp., Francisella tularensis and general Anaplasmataceae. A droplet digital PCR assay was developed for Babesia canis and A. phagocytophilum to evaluate their presence in dogs from the two populations. Pathogen prevalences between the two sample populations were compared by calculating Z-scores.

Results: Ticks were identified as Ixodes ricinus (n = 102) and Dermacentor reticulatus (n = 4). Overall, 56.9% of I. ricinus ticks were positive for at least one pathogen. A significantly higher prevalence of A. phagocytophilum and B. burgdorferi was found in ticks at the NPP (44.0% and 42.0%, respectively) compared to CC (23.1% and 19.2%, respectively). Babesia spp. (including B. canis and B. caballi) were detected in 8.8% ticks at similar proportions for both populations. Interestingly, we found a significantly lower level of A. phagocytophilum in dogs at the NPP (1.8%) than in dogs at CC (11.7%). In total, 24.3% of dogs were positive for B. canis, evenly distributed across the two populations.

Conclusions: The results of this study show contrasting pathogen prevalence in both ticks and dogs at the NPP and CC, which may reflect the differential exposures at the two locations. This work adds an important new component to our understanding of the consequences of prolonged exposure to environmental contamination on the wildlife and ecology within the Chornobyl Exclusion Zone.

背景:1986 年切尔诺贝利核电站灾难向当地环境释放了大量放射性物质。除辐射外,修复工作和军工综合体的废弃也造成了重金属、有机物、杀虫剂和其他有毒化学品的污染。许多研究评估了这种污染对当地生态环境的影响。然而,很少有研究报道这种污染对病媒传播的病原体及其宿主的影响。在本手稿中,我们描述了切尔诺贝利禁区内两个采样点的蜱媒病原体存在情况,其中一个采样点位于核电站(NPP),另一个采样点位于 16 公里外的切尔诺贝利市(CC):方法:从核电站和切尔诺贝利城捕获的自由繁殖犬身上采集蜱虫和全血样本。采用终点 PCR 和定量 PCR 方法鉴定蜱的种类,并评估是否存在特定的蜱媒病原体,包括噬细胞阿纳疟原虫、晚期鲍曼不动杆菌、巴贝斯菌属、巴顿菌属、土拉弗氏菌和一般阿纳疟原虫。针对犬巴贝斯虫和噬细胞甲虫开发了一种液滴数字 PCR 检测方法,以评估这两种病原体在两个种群的狗体内的存在情况。通过计算 Z 值,比较了两个样本人群的病原体流行率:结果:蜱虫被鉴定为蓖麻弓形虫(Ixodes ricinus)(n = 102)和皮蝇(Dermacentor reticulatus)(n = 4)。总体而言,56.9%的蓖麻蜱对至少一种病原体呈阳性。与CC(分别为23.1%和19.2%)相比,在NPP(分别为44.0%和42.0%)的蜱虫中发现的噬菌体和勃氏菌的流行率明显更高。在两个种群中,8.8%的蜱虫中检测到巴贝西亚原虫(包括犬科巴贝西亚原虫和卡巴莱巴贝西亚原虫),比例相似。有趣的是,我们在 NPP(1.8%)的犬只中发现的噬细胞甲虫数量明显低于 CC(11.7%)的犬只。总共有 24.3% 的犬对犬噬菌体呈阳性反应,均匀地分布在两个种群中:这项研究的结果表明,蜱虫和狗在 NPP 和 CC 中的病原体流行率形成了鲜明对比,这可能反映了两地接触病原体的不同情况。这项工作为我们了解长期暴露于环境污染对切尔诺贝利禁区内野生动物和生态的影响增添了重要的新内容。
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