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Comparison of multi-parallel quantitative real-time PCRs targeting different DNA regions and detecting soil-transmitted helminths in stool 针对不同 DNA 区域的多平行定量实时 PCR 与检测粪便中土壤传播蠕虫的比较

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-13 DOI: 10.1186/s13071-024-06464-6

Marina Papaiakovou, Rubén O. Cimino, Nils Pilotte, Julia Dunn, D. Timothy J. Littlewood, Steven A. Williams, Alejandro J. Krolewiecki, Rojelio Mejia

Soil-transmitted helminths infect an estimated 18% of the world’s population, causing a significant health burden. Microscopy has been the primary tool for diagnosing eggs from fecal samples, but its sensitivity drops in low-prevalence settings. Quantitative real-time polymerase chain reaction (qPCR) is slowly increasing in research and clinical settings. However, there is still no consensus on preferred qPCR targets. We aimed to compare soil-transmitted helminth (STH) DNA detection methods by testing naïve stool samples spiked with known quantities of STH eggs and larvae. DNA extracts from spiked samples were tested using independent quantitative realtime PCR (qPCR) assays targeting ribosomal or putative non-protein coding satellite sequences. For Trichuris trichiura, there was a strong correlation between egg/larvae counts and qPCR results using either qPCR method (0.86 and 0.87, respectively). Strong correlations also existed for A. lumbricoides (0.60 and 0.63, respectively), but weaker correlations were found for Ancylostoma duodenale (0.41 for both assays) and Strongyloides stercoralis (0.48 and 0.65, respectively). No correlation for Necator americanus was observed when testing with either qPCR assay. Both assays had fair-to-moderate agreement across targets when using field-collected stool samples (0.28–0.45, for all STHs), except for S. stercoralis (0.12) with slight agreement. There is a strong correlation between qPCR results and egg/larvae counts. Our study confirms that qPCR is an effective diagnostic tool, even with low-intensity infections, regardless of the DNA-based diagnostic marker used. However, the moderate agreement between the two different qPCR assays when testing field samples highlights the need to understand the role of these targets in the genome so that the parasite burden can be quantified more accurately and consistently by qPCR.

据估计，土壤传播的蠕虫感染了全球 18% 的人口，造成了巨大的健康负担。显微镜一直是诊断粪便样本中虫卵的主要工具，但在低发病率环境下，其灵敏度会下降。实时定量聚合酶链反应（qPCR）在研究和临床中的应用正在缓慢增加。然而，人们对首选的 qPCR 目标仍未达成共识。我们的目的是通过检测添加了已知数量 STH 虫卵和幼虫的天真粪便样本，比较土壤传播蠕虫（STH）DNA 检测方法。针对核糖体或假定的非蛋白编码卫星序列，我们使用独立的定量实时 PCR（qPCR）检测方法对加标样本的 DNA 提取物进行了检测。对于毛滴虫，使用任一种 qPCR 方法检测的虫卵/幼虫计数与 qPCR 结果之间都有很强的相关性（分别为 0.86 和 0.87）。疟原虫的相关性也很强（分别为 0.60 和 0.63），但十二指肠疟（两种检测方法的相关性均为 0.41）和盘尾丝虫病（分别为 0.48 和 0.65）的相关性较弱。在使用两种 qPCR 检测方法进行检测时，均未观察到与美洲鼻疽的相关性。在使用现场采集的粪便样本时，这两种检测方法在不同目标上都有相当到中等程度的一致性（0.28-0.45，适用于所有 STHs），只有盘尾丝虫（0.12）略有一致性。qPCR 结果与虫卵/幼虫计数之间有很强的相关性。我们的研究证实，无论使用哪种基于 DNA 的诊断标记，qPCR 都是一种有效的诊断工具，即使是低强度感染。不过，在检测现场样本时，两种不同的 qPCR 检测方法之间的一致性一般，这突出表明有必要了解这些靶标在基因组中的作用，以便通过 qPCR 更准确、更一致地量化寄生虫负担。

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引用次数: 0

Report of a new species of sand fly, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), from Western Ghats, India 报告来自印度西高止山脉的沙蝇新品种 Phlebotomus (Anaphlebotomus) ajithii n. sp.

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-12 DOI: 10.1186/s13071-024-06468-2

Harish Kumar Shah, P. A. Fathima, Jose Jicksy, Prasanta Saini

Western Ghats is a biodiversity treasure trove with reports of indigenous leishmaniasis cases. Hence, systematic sand fly surveillance was carried out among the tribal population. The present study reports a novel sand fly species, Phlebotomus (Anaphlebotomus) ajithii n. sp. (Diptera: Psychodidae), discovered in the Western Ghats of India. A comprehensive sand fly survey was conducted across the Kollam, Thrissur, Idukki, Kasaragod and Malappuram districts of Kerala, India. The survey spanned both indoor and outdoor habitats using standard collection methods over a 3-year, 3-month period. DNA barcoding of samples was performed targeting mitochondrial cytochrome c oxidase subunit I (COI) gene, and the sequence generated was subjected to phylogenetic analysis. Phlebotomus (Anaphlebotomus) ajithii, a new sand fly species, is recorded and described in this communication. The morphological relationship of the new species to other members of the subgenus Anaphlebotomus is discussed. Mitochondrial COI barcode followed by phylogenetic analysis confirmed that specimens of Ph. ajithii belong to the same taxonomic group, while a genetic distance of 11.7% from congeners established it as a distinct species. The Western Ghats, known for its rich biodiversity, has lacked systematic entomological surveys focusing on sand flies. This study aims to fill this gap and reports and describes a new species of sand fly.

西高止山脉是一个生物多样性宝库，据报告有本地利什曼病病例。因此，在部落人口中开展了系统的沙蝇监测。本研究报告了在印度西高止山发现的一种新型沙蝇--Phlebotomus (Anaphlebotomus) ajithii n. sp.（双翅目：Psychodidae）。在印度喀拉拉邦的 Kollam、Thrissur、Idukki、Kasaragod 和 Malappuram 地区进行了一次全面的沙蝇调查。调查采用标准采集方法，历时 3 年 3 个月，涵盖室内和室外栖息地。针对线粒体细胞色素 c 氧化酶亚单位 I（COI）基因对样本进行了 DNA 条形编码，并对生成的序列进行了系统进化分析。本通讯记录并描述了沙蝇新种 Phlebotomus (Anaphlebotomus) ajithii。文中讨论了该新种与 Anaphlebotomus 亚属其他成员的形态学关系。线粒体 COI 条形码和系统进化分析证实 Ph. ajithii 的标本属于同一分类群，而与同属种之间 11.7% 的遗传距离则将其确定为一个独特的物种。西高止山以其丰富的生物多样性而闻名，但一直缺乏以沙蝇为重点的系统昆虫学调查。本研究旨在填补这一空白，报告并描述了沙蝇的一个新物种。

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引用次数: 0

Insights from multigene analysis: first report of a Southeast Asian Mosquito, Aedes (Mucidus) laniger (Diptera: Culicidae) on Jeju Island from Korea 多基因分析的启示：韩国济州岛上东南亚蚊子伊蚊（Mucidus）laniger（双翅目：库蚊科）的首次报告

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-12 DOI: 10.1186/s13071-024-06373-8

Woo Jun Bang, Ara Seol, Seunggwan Shin

Certain mosquitoes are known as dominant vectors worldwide, and transmit infectious diseases. The expansion of mosquito habitats due to climate change and increased human activities poses a significant health threat by facilitating the spread of various non-native infectious diseases. This study focused on the detection of the Southeast Asian mosquito species, Aedes (Mucidus) laniger (Wiedemann, 1820) on Jeju Island, the southernmost region of the Republic of Korea (ROK), highlighting the potential risks associated with the spread of vector-borne diseases, particularly emphasizing the elevated likelihood of invasion by Southeast Asian mosquitoes. Field surveys were conducted in August 2023 on Jeju Island. Adult mosquitoes were collected using BG-sentinel traps and identified to the species level using taxonomic keys. Morphological and molecular analyses were employed to confirm species designations. Molecular data, including mitochondrial and nuclear genes, were used for phylogenetic analysis, which was performed to compare and identify among recorded subgenera in ROK. Species distribution modeling for Ae. laniger was performed to predict potential habitats using R package ‘BIOMOD2’. The two specimens of Ae. laniger were collected for the first time on Jeju Island. Morphological and molecular analyses confirmed the identity of this species within the subgenus Mucidus and validated the first record of this species in the ROK. We employed a simple multigene phylogenetic analysis to confirm a new mosquito record at the genus and subgenus levels, finally validating the consistency between morphological identification and molecular phylogenetic outcomes. Furthermore, we have updated the taxonomic keys for the genus Aedes in the ROK, and revised mosquito lists for Jeju Island, incorporating the inclusion of Ae. laniger. On the basis of species distribution modeling, the area of suitable habitat for Ae. laniger is expected to expand due to climate change, but this change did not appear to be meaningful in East Asia. This case offers the first report of the Southeast Asian mosquito, Ae. laniger, in the ROK. The detection of this species on Jeju Island suggests the potential establishment of a breeding population their habitat and raises concerns about further expansion into the Korean Peninsula. Considering the annual occurrence of mosquito-borne disease cases in the Southeast Asia, it is essential to conduct monitoring not only in Jeju Island, where Ae. laniger has been identified, but also across the entire Korean Peninsula.

众所周知，某些蚊子是全球主要的传播媒介，传播传染病。由于气候变化和人类活动的增加，蚊子的栖息地扩大，促进了各种非本地传染病的传播，对健康构成了严重威胁。本研究重点是在大韩民国（韩国）最南端的济州岛上发现东南亚蚊子物种伊蚊（Mucidus）laniger（Wiedemann，1820 年），突出了与病媒传播疾病有关的潜在风险，特别强调了东南亚蚊子入侵的可能性增加。2023 年 8 月在济州岛进行了实地调查。使用 BG 哨兵诱捕器收集成蚊，并使用分类钥匙进行物种鉴定。采用形态学和分子分析确认物种名称。分子数据（包括线粒体和核基因）被用于系统发育分析，以比较和识别韩国记录的亚属。利用 R 软件包 "BIOMOD2 "对兰氏疟蚊进行物种分布建模，以预测潜在的栖息地。首次在济州岛采集到两只Ae.形态学和分子分析证实了该物种在 Mucidus 亚属中的身份，并验证了该物种在韩国的首次记录。我们采用简单的多基因系统发育分析，在属和亚属水平上确认了新的蚊子记录，最终验证了形态鉴定和分子系统发育结果之间的一致性。此外，我们还更新了韩国伊蚊属的分类钥匙，并修订了济州岛的蚊子名单，将兰氏伊蚊纳入其中。根据物种分布模型，兰尼格伊蚊的适宜栖息地面积预计将因气候变化而扩大，但这一变化在东亚似乎并无意义。本案例是韩国首次报告东南亚蚊子 Ae. laniger。在济州岛发现该物种表明，其栖息地有可能建立起一个繁殖种群，并引发了对其进一步向朝鲜半岛扩张的担忧。考虑到东南亚地区每年都会发生由蚊子传播的疾病病例，因此不仅要在济州岛（已发现兰尼格伊蚊），而且要在整个朝鲜半岛进行监测。

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引用次数: 0

Baseline gut microbiota diversity and composition and albendazole efficacy in hookworm-infected individuals 钩虫感染者的基线肠道微生物群多样性和组成以及阿苯达唑的疗效

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-12 DOI: 10.1186/s13071-024-06469-1

Javier Gandasegui, Pedro E. Fleitas, Paula Petrone, Berta Grau-Pujol, Valdemiro Novela, Elisa Rubio, Osvaldo Muchisse, Anélsio Cossa, José Carlos Jamine, Charfudin Sacoor, Eric A. T. Brienen, Lisette van Lieshout, José Muñoz, Climent Casals-Pascual

Soil-transmitted helminth (STH) infections account for a significant global health burden, necessitating mass drug administration with benzimidazole-class anthelmintics, such as albendazole (ALB), for morbidity control. However, ALB efficacy shows substantial variability, presenting challenges for achieving consistent treatment outcomes. We have explored the potential impact of the baseline gut microbiota on ALB efficacy in hookworm-infected individuals through microbiota profiling and machine learning (ML) techniques. Our investigation included 89 stool samples collected from hookworm-infected individuals that were analyzed by microscopy and quantitative PCR (qPCR). Of these, 44 were negative by microscopy for STH infection using the Kato-Katz method and qPCR 21 days after treatment, which entails a cure rate of 49.4%. Microbiota characterization was based on amplicon sequencing of the V3–V4 16S ribosomal RNA gene region. Alpha and beta diversity analyses revealed no significant differences between participants who were cured and those who were not cured, suggesting that baseline microbiota diversity does not influence ALB treatment outcomes. Furthermore, differential abundance analysis at the phylum, family and genus levels yielded no statistically significant associations between bacterial communities and ALB efficacy. Utilizing supervised ML models failed to predict treatment response accurately. Our investigation did not provide conclusive insights into the relationship between gut microbiota and ALB efficacy. However, the results highlight the need for future research to incorporate longitudinal studies that monitor changes in the gut microbiota related to the infection and the cure with ALB, as well as functional metagenomics to better understand the interaction of the microbiome with the drug, and its role, if there is any, in modulating anthelmintic treatment outcomes in STH infections. Interdisciplinary approaches integrating microbiology, pharmacology, genetics and data science will be pivotal in advancing our understanding of STH infections and optimizing treatment strategies globally.

土壤传播的蠕虫（STH）感染给全球健康造成了巨大负担，因此有必要大规模使用苯并咪唑类抗虫药，如阿苯咪唑（ALB），以控制发病率。然而，阿苯达唑的疗效存在很大的变异性，给实现一致的治疗效果带来了挑战。我们通过微生物群分析和机器学习（ML）技术探讨了基线肠道微生物群对钩虫感染者的 ALB 疗效的潜在影响。我们的调查包括从钩虫感染者身上采集的 89 份粪便样本，并通过显微镜和定量 PCR (qPCR) 进行了分析。其中，44 份样本在治疗 21 天后通过卡托-卡茨法显微镜检查和 qPCR 检测为阴性，治愈率为 49.4%。微生物群的特征描述基于 V3-V4 16S 核糖体 RNA 基因区的扩增片段测序。阿尔法和贝塔多样性分析表明，治愈者与未治愈者之间没有显著差异，这表明基线微生物群多样性不会影响 ALB 的治疗效果。此外，在门、科和属一级进行的丰度差异分析显示，细菌群落与 ALB 疗效之间没有统计学意义上的关联。利用有监督的 ML 模型无法准确预测治疗反应。我们的调查并未就肠道微生物群与 ALB 药效之间的关系提供结论性见解。不过，研究结果突出表明，未来的研究需要纳入纵向研究，监测与感染和 ALB 治疗相关的肠道微生物群变化，并纳入功能元基因组学研究，以更好地了解微生物群与药物之间的相互作用，以及微生物群在调节 STH 感染中抗虫药治疗效果方面的作用（如果有的话）。整合微生物学、药理学、遗传学和数据科学的跨学科方法将在促进我们对性传播疾病感染的了解和优化全球治疗策略方面发挥关键作用。

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引用次数: 0

A study on the diversity of phlebotomine sand flies (Diptera, Psychodidae) in karstic limestone areas in Vientiane Province, Laos, with a description of two new species of Sergentomyia França and & Parrot 老挝万象岩溶石灰岩地区沙蝇（双翅目，灵蝇科）的多样性研究，并描述了 Sergentomyia França 和 & Parrot 的两个新种

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-11 DOI: 10.1186/s13071-024-06444-w

Khamsing Vongphayloth, Fano José Randrianambinintsoa, Khaithong Lakeomany, Nothasine Phommavanh, Tavun Pongsanarm, Veaky Vungkyly, Phonesavanh Luangamath, Somsanith Chonephetsarath, Paul T. Brey, Jérôme Depaquit

Southeast Asia is well known as a hotspot of biodiversity. However, very little is known about cave-dwelling hematophagous insects that are medically important. Taxonomic knowledge and ecology of phlebotomine sand flies are very poorly studied in Laos, as well as in other countries in the region. Herein, we report species diversity data and some notes on the ecology of the detected species from these karstic limestone areas of Laos. Phlebotomine sand flies were collected using Centers for Disease Control and Prevention (CDC) light traps from limestone cave locations in three districts of Vientiane Province, Laos. Both morphological and molecular techniques were used for sand fly identification. Species diversity and abundance were analyzed according to sites, locations, collection seasons, and trapping positions. A total of 6564 sand flies, of which 5038 were females and 1526 were males, were morphologically identified into 20 species belonging to five genera (Chinius, Idiophlebotomus, Phlebotomus, Sergentomyia, and Grassomyia). The most abundant species were Chinius eunicegalatiae, Phlebotomus stantoni, Sergentomyia hivernus, Se. siamensis, and Idiophlebotomus longiforceps. Cytochrome b analysis results supported the morphological identification and revealed that Se. siamensis was separated from other members of the Se. barraudi group. Two new species, Se. dvoraki n. sp. and Se. marolii n. sp., were described. Sand fly density was generally high except in a cave in Vangvieng, with species richness ranging from 14 to 18 across different caves. Outside caves had higher species richness (R = 20) and diversity (H = 2.50) than cave entrances (R = 18, H = 2.41) and interiors (R = 16, H = 2.13). Seasonal variations showed high sand fly density in Feung and Hinheup during both dry and rainy seasons, while Vangvieng had a notable decrease in density during the dry season (D = 6.29). This study revealed that the diversity of phlebotomine sand fly fauna in Laos, particularly in karstic limestone areas, is greater than previously known. However, the taxonomic status of many species in Laos, as well as Southeast Asia, still needs more in-depth study using both morphological characters and molecular methods. Many species could be found from inside, at the entrance, and outside of caves, indicating a wide range of host-seeking behavior or possible natural breeding in the karstic cave areas.

众所周知，东南亚是生物多样性的热点地区。然而，人们对具有重要医学价值的穴居食血昆虫知之甚少。老挝以及该地区其他国家对血沙蝇的分类学知识和生态学研究非常少。在此，我们报告了老挝喀斯特石灰岩地区发现的物种多样性数据和一些生态学说明。我们使用美国疾病控制和预防中心（CDC）的灯光诱捕器从老挝万象省三个地区的石灰岩洞穴中收集了沙蝇。采用形态学和分子技术对沙蝇进行鉴定。根据地点、位置、采集季节和诱捕位置对物种多样性和丰度进行了分析。经形态学鉴定，共有 6564 只沙蝇，其中 5038 只为雌蝇，1526 只为雄蝇，隶属于 5 个属（Chinius 属、Idiophlebotomus 属、Phlebotomus 属、Sergentomyia 属和 Grassomyia 属）的 20 个种。数量最多的物种是 Chinius eunicegalatiae、Phlebotomus stantoni、Sergentomyia hivernus、Se. siamensis 和 Idiophlebotomus longiforceps。细胞色素 b 分析结果支持形态学鉴定，并表明 Se. siamensis 与 Se. barraudi 群的其他成员是分开的。描述了两个新种：Se. dvoraki n. sp.和 Se. marolii n. sp.。除万维翁的一个洞穴外，其他洞穴的沙蝇密度普遍较高，不同洞穴的沙蝇物种丰富度在 14 至 18 种之间。洞穴外的物种丰富度（R = 20）和多样性（H = 2.50）高于洞穴入口（R = 18，H = 2.41）和洞穴内部（R = 16，H = 2.13）。季节变化显示，在旱季和雨季，Feung 和 Hinheup 的沙蝇密度都很高，而 Vangvieng 的沙蝇密度在旱季明显下降（D = 6.29）。这项研究表明，老挝（尤其是喀斯特石灰岩地区）噬血沙蝇动物群的多样性比以前已知的要高。然而，老挝乃至东南亚许多物种的分类地位仍需要利用形态特征和分子方法进行更深入的研究。许多物种在洞内、洞口和洞外都能找到，这表明它们在喀斯特洞穴地区有广泛的寻找寄主行为或可能的自然繁殖。

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引用次数: 0

Real-time PCR for malaria diagnosis and identification of Plasmodium species in febrile patients in Cubal, Angola 用实时 PCR 技术诊断疟疾并鉴定安哥拉库巴尔发热病人体内的疟原虫种类

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-11 DOI: 10.1186/s13071-024-06467-3

Alejandro Mediavilla, Aroa Silgado, Begoña Febrer-Sendra, Beatriz Crego-Vicente, Patricia Martínez-Vallejo, Carles Rubio Maturana, Lidia Goterris, Arlette Nindia, Joan Martínez-Campreciós, Sandra Aixut, María Luisa Aznar-Ruiz-de-Alegría, Pedro Fernández-Soto, Antonio Muro, Fernando Salvador, Israel Molina, Pedro Berzosa, Inés Oliveira-Souto, Elena Sulleiro

Malaria is the parasitic disease with the highest morbimortality worldwide. The World Health Organization (WHO) estimates that there were approximately 249 million cases in 2022, of which 3.4% were in Angola. Diagnosis is based on parasite identification by microscopy examination, antigen detection, and/or molecular tests, such as polymerase chain reaction (PCR). This study aimed to evaluate the usefulness of real-time PCR as a diagnostic method for malaria in an endemic area (Cubal, Angola). A cross-sectional study was carried out at the Hospital Nossa Senhora da Paz in Cubal, Angola, including 200 patients who consulted for febrile syndrome between May and July 2022. From each patient, a capillary blood sample was obtained by finger prick for malaria field diagnosis [microscopy and rapid diagnostic test (RDT)] and venous blood sample for real-time PCR performed at the Hospital Universitario Vall d’Hebron in Barcelona, Spain. Any participant with a positive result from at least one of these three methods was diagnosed with malaria. Of the 200 participants included, 54% were female and the median age was 7 years. Malaria was diagnosed by at least one of the three techniques (microscopy, RDT, and/or real-time PCR) in 58% of the participants, with RDT having the highest percentage of positivity (49%), followed by real-time PCR (39.5%) and microscopy (33.5%). Of the 61 discordant samples, 4 were only positive by microscopy, 13 by real-time PCR, and 26 by RDT. Plasmodium falciparum was the most frequent species detected (90.63%), followed by P. malariae (17.19%) and P. ovale (9.38%). Coinfections were detected in ten participants (15.63%): six (60%) were caused by P. falciparum and P. malariae, three (30%) by P. falciparum and P. ovale, and one (10%) triple infection with these three species. In addition, it was observed that P. falciparum and P. malariae coinfection significantly increased the parasite density of the latter. RDT was the technique with the highest positivity rate, followed by real-time PCR and microscopy. The results of the real-time PCR may have been underestimated due to suboptimal storage conditions during the transportation of the DNA eluates. However, real-time PCR techniques have an important role in the surveillance of circulating Plasmodium species, given the epidemiological importance of the increase in non-falciparum species in the country, and can provide an estimate of the intensity of infection.

疟疾是全世界死亡率最高的寄生虫病。世界卫生组织（WHO）估计，2022年约有2.49亿病例，其中3.4%发生在安哥拉。诊断的依据是通过显微镜检查、抗原检测和/或聚合酶链反应（PCR）等分子检测来识别寄生虫。本研究旨在评估实时聚合酶链反应作为疟疾流行地区（安哥拉库巴尔）疟疾诊断方法的实用性。这项横断面研究在安哥拉库巴尔的帕斯圣母医院（Hospital Nossa Senhora da Paz）进行，包括2022年5月至7月期间因发热综合征就诊的200名患者。每位患者的毛细血管血样均由西班牙巴塞罗那瓦尔德希布伦大学医院采集，用于疟疾现场诊断（显微镜检查和快速诊断检测（RDT））和静脉血样本的实时 PCR 检测。凡是在这三种方法中至少有一种方法检测结果呈阳性的参与者均被诊断为疟疾患者。在 200 名参与者中，54% 为女性，年龄中位数为 7 岁。58% 的参与者至少通过三种技术（显微镜检查、RDT 和/或实时 PCR）中的一种诊断出疟疾，其中 RDT 阳性率最高（49%），其次是实时 PCR（39.5%）和显微镜检查（33.5%）。在 61 份不一致的样本中，4 份仅显微镜检查呈阳性，13 份为实时 PCR 阳性，26 份为 RDT 阳性。恶性疟原虫是最常检测到的疟原虫种类（90.63%），其次是疟疾疟原虫（17.19%）和卵形疟原虫（9.38%）。在 10 名参与者（15.63%）中发现了合并感染：6 人（60%）由恶性疟原虫和疟疾疟原虫引起，3 人（30%）由恶性疟原虫和卵形疟原虫引起，1 人（10%）同时感染了这三种疟原虫。此外，还观察到恶性疟原虫和疟疾疟原虫混合感染会显著增加后者的寄生虫密度。RDT 是阳性率最高的技术，其次是实时 PCR 和显微镜检查。实时 PCR 的结果可能被低估了，原因是 DNA 洗脱液在运输过程中的储存条件不够理想。不过，考虑到该国非疟原虫种类增加在流行病学上的重要性，实时 PCR 技术在监测循环疟原虫种类方面具有重要作用，并可提供感染强度的估计值。

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引用次数: 0

Mosquito dynamics and their drivers in peri-urban Antananarivo, Madagascar: insights from a longitudinal multi-host single-site survey 马达加斯加塔那那利佛近郊的蚊虫动态及其驱动因素：多宿主单点纵向调查的启示

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-10 DOI: 10.1186/s13071-024-06393-4

Michaël Luciano Tantely, Hélène Guis, Manou Rominah Raharinirina, Maminirina Fidelis Ambinintsoa, Iavonirina Randriananjantenaina, Haja Johnson Velonirina, Christophe Revillion, Vincent Herbreteau, Annelise Tran, Romain Girod

Antananarivo, the capital city of Madagascar, is experiencing a steady increase in population growth. Due to the abundance of mosquito vectors in this locality, the population exposed to mosquito-borne diseases is therefore also increasing, as is the risk of epidemic episodes. The aim of the present study was to assess, in a resource-limited setting, the information on mosquito population dynamics and disease transmission risk that can be provided through a longitudinal entomological study carried out in a multi-host single site. Mosquitoes were collected every 15 days over 16 months (from January 2017 to April 2018) using six CDC-light traps in a peri-urban area of Antananarivo. Multivariable generalised linear models were developed using indoor and outdoor densities of the predominant mosquito species as response variables and moon illumination, environmental data and climatic data as the explanatory variables. Overall, 46,737 mosquitoes belonging to at least 20 species were collected, of which Culex antennatus (68.9%), Culex quinquefasciatus (19.8%), Culex poicilipes (3.7%) and Anopheles gambiae sensu lato (2.3%) were the most abundant species. Mosquito densities were observed to be driven by moon illumination and climatic factors interacting at different lag periods. The outdoor models demonstrated biweekly and seasonal patterns of mosquito densities, while the indoor models demonstrated only a seasonal pattern. An important diversity of mosquitoes exists in the peri-urban area of Antananarivo. Some well-known vector species, such as Cx. antennatus, a major vector of West Nile virus (WNV) and Rift-Valley fever virus (RVFV), Cx. quinquefasciatus, a major vector of WNV, Cx. poicilipes, a candidate vector of RVFV and An. gambiae sensu lato, a major vector of Plasmodium spp., are abundant. Importantly, these four mosquito species are present all year round, even though their abundance declines during the cold dry season, with the exception of Cx. quinquefasciatus. The main drivers of their abundance were found to be temperature, relative humidity and precipitation, as well as—for outdoor abundance only—moon illumination. Identifying these drivers is a first step towards the development of pathogen transmission models (R0 models), which are key to inform public health stakeholders on the periods of most risk for vector-borne diseases.

马达加斯加首都塔那那利佛的人口正在稳步增长。由于该地区蚊虫载体数量众多，因此受蚊虫传播疾病影响的人口也在增加，流行病爆发的风险也在增加。本研究的目的是在资源有限的情况下，通过在多宿主单一地点进行纵向昆虫学研究，评估蚊子种群动态和疾病传播风险方面的信息。在 16 个月内（2017 年 1 月至 2018 年 4 月），在塔那那利佛的一个近郊区使用 6 个 CDC 灯光诱捕器每 15 天收集一次蚊子。以主要蚊子物种的室内和室外密度为响应变量，以月亮光照、环境数据和气候数据为解释变量，建立了多变量广义线性模型。研究共收集到 46 737 隻蚊子，分屬最少 20 個品種，其中以庫蚊（68.9%）、五節纏繞庫蚊（19.8%）、櫛纏繞庫蚊（3.7%）和按蚊（2.3%）最多。据观察，蚊子密度受不同滞后期的月照和气候因素相互作用的影响。室外模型显示了蚊子密度的双周和季节模式，而室内模型只显示了季节模式。塔那那利佛近郊区的蚊子种类繁多。一些众所周知的病媒物种，如西尼罗河病毒（WNV）和裂谷热病毒（RVFV）的主要病媒 Cx. antennatus、WNV 的主要病媒 Cx. quinquefasciatus、RVFV 的候选病媒 Cx. poicilipes 和疟原虫的主要病媒 An. gambiae sensu lato 都大量存在。重要的是，这四种蚊子全年都存在，尽管在寒冷的旱季它们的数量会减少，但五步蛇除外。研究发现，温度、相对湿度和降水量是影响蚊子数量的主要因素，而对于室外蚊子来说，月光也是影响蚊子数量的主要因素。确定这些驱动因素是开发病原体传播模型（R0 模型）的第一步，该模型是让公共卫生利益相关者了解病媒传播疾病最危险时期的关键。

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引用次数: 0

Molecular xenomonitoring as an indicator of microfilaraemia prevalence for lymphatic filariasis in Samoa in 2019 将分子异种监测作为 2019 年萨摩亚淋巴丝虫病微丝蚴血症流行率的指标

IF 3.2 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-09 DOI: 10.1186/s13071-024-06463-7

Maddison Howlett, Helen J. Mayfield, Brady McPherson, Lisa Rigby, Robert Thomsen, Steven A. Williams, Nils Pilotte, Shannon M. Hedtke, Patricia M. Graves, Therese Kearns, Take Naseri, Sarah Sheridan, Angus McLure, Colleen L. Lau

Lymphatic filariasis (LF) is a globally significant, vector-borne, neglected tropical disease that can result in severe morbidity and disability. As the World Health Organization (WHO) Global Programme to Eliminate Lymphatic Filariasis makes progress towards LF elimination, there is greater need to develop sensitive strategies for post-intervention surveillance. Molecular xenomonitoring (MX), the detection of pathogen DNA in vectors, may provide a sensitive complement to traditional human-based surveillance techniques, including detection of circulating filarial antigen and microfilaraemia (Mf). This study aims to explore the relationship between human Mf prevalence and the prevalence of polymerase chain reaction (PCR)-positive mosquitoes using MX. This study compared Mf and MX results from a 2019 community-based survey conducted in 35 primary sampling units (PSUs) in Samoa. This study also investigated concordance between presence and absence of PCR-positive mosquitoes and Mf-positive participants at the PSU level, and calculated sensitivity and negative predictive values for each indicator using presence of any Mf-positive infection in humans or PCR-positive mosquitoes as a reference. Correlation between prevalence of filarial DNA in mosquitoes and Mf in humans was estimated at the PSU and household/trap level using mixed-effect Bayesian multilevel regression analysis. Mf-positive individuals were identified in less than half of PSUs in which PCR-positive mosquito pools were present (13 of 28 PSUs). Prevalence of PCR-positive mosquitoes (each species separately) was positively correlated with Mf prevalence in humans at the PSU level. Analysed at the species level, only Aedes polynesiensis demonstrated strong evidence of positive correlation (r) with human Mf prevalence at both PSU (r: 0.5, 95% CrI 0.1–0.8) and trap/household levels (r: 0.6, 95% CrI 0.2–0.9). Findings from this study demonstrate that MX can be a sensitive surveillance method for identifying residual infection in low Mf prevalence settings. MX identified more locations with signals of transmission than Mf-testing. Strong correlation between estimated PCR-positive mosquitoes in the primary vector species and Mf in humans at small spatial scales demonstrates the utility of MX as an indicator for LF prevalence in Samoa and similar settings. Further investigation is needed to develop MX guidelines to strengthen the ability of MX to inform operational decisions.

淋巴丝虫病（LF）是一种全球性的病媒传染的被忽视热带疾病，可导致严重的发病率和残疾。随着世界卫生组织（WHO）消除淋巴丝虫病全球计划在消除淋巴丝虫病方面取得进展，我们更需要制定敏感的干预后监测战略。分子异种监测（MX）是对载体中病原体DNA的检测，可为传统的人体监测技术（包括检测循环丝虫抗原和微丝蚴血症（Mf））提供灵敏的补充。本研究旨在利用 MX 技术探讨人类 Mf 感染率与聚合酶链反应（PCR）阳性蚊子感染率之间的关系。本研究比较了在萨摩亚 35 个初级采样单位（PSU）进行的 2019 年社区调查的 Mf 和 MX 结果。本研究还调查了PSU水平上是否存在PCR阳性蚊子和Mf阳性参与者之间的一致性，并以是否存在任何Mf阳性的人类感染或PCR阳性蚊子作为参考，计算了每个指标的灵敏度和阴性预测值。利用混合效应贝叶斯多层次回归分析法，在PSU和家庭/诱捕器层面估算了蚊子中丝虫DNA流行率与人类Mf流行率之间的相关性。在存在PCR阳性蚊子池的PSU中，只有不到一半（28个PSU中的13个）能发现Mf阳性个体。在 PSU 层面上，PCR 阳性蚊子的流行率（每个物种单独计算）与人类 Mf 流行率呈正相关。在物种水平上进行分析，只有波利尼西亚伊蚊在 PSU（r：0.5，95% CrI 0.1-0.8）和诱捕器/住户水平（r：0.6，95% CrI 0.2-0.9）上与人类 Mf 感染率有很强的正相关性（r）。这项研究的结果表明，MX 可以作为一种灵敏的监测方法，用于识别低 Mf 流行率环境中的残余感染。与 Mf 检测相比，MX 能识别出更多有传播信号的地点。在较小的空间范围内，主要病媒物种中估计的 PCR 阳性蚊子与人类中的甲沟炎之间存在很强的相关性，这表明在萨摩亚和类似的环境中，MX 作为甲沟炎流行指标是有用的。需要进一步调查，以制定 MX 指南，加强 MX 为业务决策提供信息的能力。

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引用次数: 0

Trypanosoma cruzi reprograms mitochondrial metabolism within the anterior midgut of its vector Rhodnius prolixus during the early stages of infection. 在感染的早期阶段，克鲁斯锥虫对其载体 Rhodnius prolixus 的前中肠内的线粒体代谢进行了重编程。

IF 3 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-06 DOI: 10.1186/s13071-024-06415-1

Radouane Ouali, Larissa Rezende Vieira, Didier Salmon, Sabrina Bousbata

Background: Trypanosoma cruzi is transmitted to humans by hematophagous bugs belonging to the Triatominae subfamily. Its intra-vectorial cycle is complex and occurs exclusively in the insect's midgut. Dissecting the elements involved in the cross-talk between the parasite and its vector within the digestive tract should provide novel targets for interrupting the parasitic life cycle and affecting vectorial competence. These interactions are shaped by the strategies that parasites use to infect and exploit their hosts, and the host's responses that are designed to detect and eliminate parasites. The objective of the current study is to characterize the impact of T. cruzi establishment within its vector on the dynamics of its midgut.

Methods: In this study, we evaluated the impact of T. cruzi infection on protein expression within the anterior midgut of the model insect Rhodnius prolixus at 6 and 24 h post-infection (hpi) using high-throughput quantitative proteomics.

Results: Shortly after its ingestion, the parasite modulates the proteome of the digestive epithelium by upregulating 218 proteins and negatively affecting the expression of 11 proteins involved in a wide array of cellular functions, many of which are pivotal due to their instrumental roles in cellular metabolism and homeostasis. This swift response underscores the intricate manipulation of the vector's cellular machinery by the parasite. Moreover, a more in-depth analysis of proteins immediately induced by the parasite reveals a pronounced predominance of mitochondrial proteins, thereby altering the sub-proteomic landscape of this organelle. This includes various complexes of the respiratory chain involved in ATP generation. In addition to mitochondrial metabolic dysregulation, a significant number of detoxifying proteins, such as antioxidant enzymes and P450 cytochromes, were immediately induced by the parasite, highlighting a stress response.

Conclusions: This study is the first to illustrate the response of the digestive epithelium upon contact with T. cruzi, as well as the alteration of mitochondrial sub-proteome by the parasite. This manipulation of the vector's physiology is attributable to the cascade activation of a signaling pathway by the parasite. Understanding the elements of this response, as well as its triggers, could be the foundation for innovative strategies to control the transmission of American trypanosomiasis, such as the development of targeted interventions aimed at disrupting parasite proliferation and transmission within the triatomine vector.

背景：克鲁兹锥虫是由三足虫亚科的食血虫传播给人类的。它的载体内循环非常复杂，而且只发生在昆虫的中肠。剖析寄生虫与其载体在消化道内交叉对话所涉及的要素，将为中断寄生虫生命周期和影响载体能力提供新的目标。寄生虫感染和利用宿主的策略以及宿主检测和消灭寄生虫的反应决定了这些相互作用。本研究的目的是确定在载体内建立 T. cruzi 对其中肠道动态的影响：在这项研究中，我们利用高通量定量蛋白质组学评估了感染 T. cruzi 对模型昆虫 Rhodnius prolixus 感染后 6 小时和 24 小时（hpi）前中肠内蛋白质表达的影响：结果：寄生虫摄入后不久就会调节消化道上皮细胞的蛋白质组，上调 218 种蛋白质，并对 11 种参与多种细胞功能的蛋白质的表达产生负面影响，其中许多蛋白质在细胞新陈代谢和稳态中起着关键作用。这种快速反应凸显了寄生虫对载体细胞机制的复杂操控。此外，对寄生虫立即诱导的蛋白质进行更深入的分析后发现，线粒体蛋白质明显占优势，从而改变了这一细胞器的亚蛋白质组格局。这包括参与产生 ATP 的呼吸链的各种复合物。除了线粒体代谢失调外，寄生虫还立即诱导了大量解毒蛋白，如抗氧化酶和 P450 细胞色素，凸显了应激反应：这项研究首次说明了消化道上皮细胞在接触克柔蚊后的反应，以及寄生虫对线粒体亚蛋白质组的改变。这种对载体生理机能的操纵可归因于寄生虫对信号通路的级联激活。了解这种反应的要素及其触发因素，可以为控制美洲锥虫病传播的创新战略奠定基础，例如制定有针对性的干预措施，以扰乱寄生虫在三蝽载体内的增殖和传播。

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引用次数: 0

Live-attenuated PruΔgra72 strain of Toxoplasma gondii induces strong protective immunity against acute and chronic toxoplasmosis in mice. 减毒的 PruΔgra72 刚地弓形虫活菌株能诱导小鼠对急性和慢性弓形虫病产生强大的保护性免疫力。

IF 3 2区医学 Q1 PARASITOLOGY

Parasites & Vectors

Pub Date : 2024-09-05 DOI: 10.1186/s13071-024-06461-9

Jing Li, Yu Kang, Ze-Xuan Wu, Shu-Feng Yang, Yu-Yang Tian, Xing-Quan Zhu, Xiao-Nan Zheng

Background: Toxoplasma gondii is an intracellular opportunistic pathogenic protozoan that poses serious threats, particularly in immunocompromised individuals. In the absence of a robust prophylactic measure, the mitigation and management of toxoplasmosis present formidable challenges to public health. We recently found that GRA72 plays an important role in parasitophorous vacuole (PV) morphology, growth and virulence of T. gondii. However, whether gra72-deficient strain can be used as a vaccine remains unknown.

Methods: We first examined the attenuated virulence of gra72 gene knockout strain (PruΔgra72) and the parasite load in organs of the infected mice. Subsequently, we evaluated the immune-protective effects of the PruΔgra72 vaccination against challenge with various types of T. gondii tachyzoites and Pru cysts. Furthermore, levels of antibodies and cytokines induced by PruΔgra72 vaccination were examined. Statistical analysis was conducted by Student's t-test or Mantel-Cox log-rank test based on data obtained from three independent experiments with GraphPad Prism 8.0.

Results: We found that PruΔgra72 strain exhibited a significantly attenuated virulence even at the highest dose of 5 × 10⁷ tachyzoites in Kunming mice model. The significant decrease of brain cyst burden and parasite load in the organs of the PruΔgra72-infected mice suggested its potentiality as a live-attenuated vaccine. Hence, we explored the protective immunity of PruΔgra72 vaccination against toxoplasmosis. Results showed that vaccination with 5 × 10⁶ PruΔgra72 tachyzoites triggered a strong and sustained Th1-biased immune response, marked by significantly increased levels of anti-T. gondii IgG antibodies, and significantly higher levels of Th1 type cytokines (IL-2, IL-12 and IFN-γ) compared to that of Th2 type (IL-4 and IL-10). Vaccination with 5 × 10⁶ PruΔgra72 tachyzoites in mice conferred long-term protection against T. gondii infection by less virulent tachyzoites (ToxoDB#9 PYS and Pru strains) and Pru cysts, provided partial protection against acute infection by high virulent Type I RH tachyzoites and significantly decreased brain cyst burden of chronically infected mice.

Conclusions: The avirulent PruΔgra72 induced strong protective immunity against acute and chronic T. gondii infection and is a promising candidate for developing a safe and effective live-attenuated vaccine against T. gondii infection.

背景：弓形虫（Toxoplasma gondii）是一种细胞内机会致病原生动物，尤其对免疫力低下的人构成严重威胁。由于缺乏强有力的预防措施，弓形虫病的缓解和管理给公共卫生带来了严峻的挑战。我们最近发现，GRA72 在弓形虫的寄生泡（PV）形态、生长和毒力方面发挥着重要作用。然而，GRA72缺陷株能否用作疫苗仍是未知数：方法：我们首先检测了gra72基因敲除株（PruΔgra72）的减毒性和感染小鼠器官中的寄生虫量。随后，我们评估了接种 PruΔgra72 疫苗对各种类型的淋病双鞭毛虫和 Pru 包囊的免疫保护作用。此外，还检测了接种 PruΔgra72 疫苗后诱导的抗体和细胞因子水平。使用 GraphPad Prism 8.0 对三个独立实验的数据进行统计分析，采用学生 t 检验或 Mantel-Cox 对数秩检验：我们发现，在昆明小鼠模型中，PruΔgra72株即使在最高剂量5×107孑孓时也表现出明显的毒力减弱。PruΔgra72感染小鼠脑囊肿负荷和寄生虫在器官中的负荷明显减少，这表明它具有作为减毒活疫苗的潜力。因此，我们探讨了接种 PruΔgra72 疫苗对弓形虫病的保护性免疫。结果表明，接种 5 × 106 PruΔgra72 支原体疫苗会引发强烈而持续的 Th1 型免疫反应，抗弓形虫 IgG 抗体水平显著提高，Th1 型细胞因子（IL-2、IL-12 和 IFN-γ）水平显著高于 Th2 型细胞因子（IL-4 和 IL-10）。给小鼠接种5×106 PruΔgra72速虫疫苗可长期保护其免受毒性较低的速虫（ToxoDB#9 PYS和Pru株）和Pru囊虫的感染，部分保护其免受毒性较高的I型RH速虫的急性感染，并显著减少慢性感染小鼠的脑囊肿负荷：结论：无病毒的PruΔgra72对急性和慢性淋病感染具有很强的保护性免疫力，是开发安全有效的淋病减毒活疫苗的理想候选菌株。

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