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Black spot diseases in seven commercial fish species from the English Channel and the North Sea: infestation levels, identification and population genetics of Cryptocotyle spp. 英吉利海峡和北海七种商业鱼类的黑斑病:隐子叶类的侵染水平、鉴定和种群遗传学。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023028
Maureen Duflot, Pierre Cresson, Maéva Julien, Léa Chartier, Odile Bourgau, Marialetizia Palomba, Simonetta Mattiucci, Graziella Midelet, Mélanie Gay

Fish are often speckled with "black spots" caused by metacercarial trematode infection, inducing a host response. Cryptocotyle spp. (Opisthorchiidae) are among the parasites responsible for this phenomenon. So far, the impact on human health is still unknown. In addition, few publications dealing with black spot recovery, identification, distribution and diversity among commercially important fish are available. Moreover, "black spots" have been observed by fishermen on marine fish, revealing an appreciable but unquantified presence in consumed fish. An epidemiological survey of 1,586 fish from seven commercial species (herring, sprat, whiting, pout, dab, flounder, and plaice) was conducted in the Eastern English Channel and the North Sea in January 2019 and 2020. Encysted metacercariae were found in 325 out of 1,586 fish, with a total prevalence of 20.5%. Intensity of infection varied from 1 to 1,104 parasites. The recorded encysted metacercariae were identified either by microscopic examination or with molecular tools. Partial sequences of the mtDNA cox1 gene and of the rDNA ITS region were obtained. Two species of Cryptocotyle, Cryptocotyle lingua (Creplin, 1825) and Cryptocotyle concava (Creplin, 1825) were found. Metacercariae belonging to other trematode families were also identified. Molecular phylogenetic analysis and haplotype network construction were performed to confirm the identification and to study the potential presence of different populations of Cryptocotyle spp. This survey enabled us to describe the distribution of two species of Cryptocotyle in the English Channel and North Sea ecosystems. The observed differences in infestation levels between fish species and geographical areas will contribute to better understanding of the ecology of these parasites.

鱼身上经常有斑点状的“黑点”,这是由包囊吸虫感染引起的,从而引起宿主的反应。隐子叶虫属(视蛾科)是造成这种现象的寄生虫之一。到目前为止,对人类健康的影响尚不清楚。此外,关于商业上重要鱼类的黑斑恢复、鉴定、分布和多样性的出版物很少。此外,渔民在海鱼身上观察到“黑点”,表明在食用的鱼中有明显但无法量化的存在。2019年1月和2020年1月,在东英吉利海峡和北海对7种商业物种(鲱鱼、鲽鱼、白鲑、鲽鱼、dab、比目鱼和鲽)的1586种鱼进行了流行病学调查。1586条鱼中有325条发现囊蚴,总患病率为20.5%。感染强度从1到1104个寄生虫不等。所记录的包囊囊蚴通过显微镜检查或分子工具进行鉴定。获得了mtDNA cox1基因和rDNA ITS区域的部分序列。发现隐子叶2种,隐子叶lingua (Creplin, 1825)和隐子叶concava (Creplin, 1825)。还发现了其他吸虫科的囊蚴。通过分子系统发育分析和单倍型网络构建,确定了两种隐子叶植物在英吉利海峡和北海生态系统中的分布情况,并探讨了隐子叶植物不同居群的存在可能性。观察到的不同鱼种和不同地理区域间侵染程度的差异将有助于更好地了解这些寄生虫的生态学。
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引用次数: 0
Three Aedes species infested by mermithids in France. 法国三种被水蚤感染的伊蚊。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023013
Jean-Philippe Martinet, Issam Aatif, Jérôme Depaquit

Mermithid nematodes (Nematoda, Mermithidae) parasitising mosquitoes could be useful as biological agents for the control of host populations. Nine female mosquitoes belonging to the species Aedes cantans, Ae. communis, and Ae. rusticus were found parasitised by mermithids in Northern France. Sequencing of partial 18S rDNA showed 100% sequence homology for all processed specimens. The mermithid sequences were closely related to specimens previously recorded from Anopheles gambiae in Senegal. However, 18S sequences do not allow for identification of nematodes at the genus or species level. Our specimens could also be related to Strelkovimermis spiculatus, or belong to other genera not yet sequenced such as Empidomermis, the only mermithid genus recorded from mosquitoes in France.

寄生于蚊虫体内的湿蛾线虫(线虫纲,湿蛾科)可作为防治蚊虫的生物制剂。广东伊蚊属9种雌蚊。共产主义和Ae。在法国北部发现了寄生在绿腹虫身上的锈蝇。部分18S rDNA测序结果显示,所有处理标本序列同源性100%。该mermithid序列与先前在塞内加尔记录的冈比亚按蚊标本密切相关。然而,18S序列不允许在属或种水平上鉴定线虫。我们的标本也可能与Strelkovimermis spiculatus有亲缘关系,或属于其他尚未测序的属,如Empidomermis,这是法国蚊子中唯一记录的mermithid属。
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引用次数: 1
Molecular detection of Cercopithifilaria, Cruorifilaria and Dipetalonema-like filarial nematodes in ticks of French Guiana. 法属圭亚那蜱虫中丝状线虫的分子检测。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023027
Florian Binetruy, Olivier Duron

Filarial nematodes of the Dipetalonema lineage are widespread parasites and include some species that are transmitted by ticks. In this study, we conducted a large molecular survey of ticks in French Guiana, South America, to understand the overall diversity of tick-borne filarioids in this remote region largely covered by dense tropical forests. Out of 682 ticks belonging to 22 species and 6 genera, 21 ticks (3.1%) of the species Amblyomma cajennense, A. oblongoguttatum, A. romitii, Ixodes luciae and Rhipicephalus sanguineus sensu lato were positive for infection by filarioids. Molecular typing and phylogenetic analysis identified all these filarioids as members of the Dipetalonema lineage. While the filarioid of R. sanguineus sensu lato is a previously described species, the canine worm Cercopithifilaria bainae Almeida & Vicente, 1984, all other filarioids detected in this study are related but distinct to already known species in the genera Cercopithifilaria, Cruorifilaria and Dipetalonema. Their vertebrate host range may include a wide variety of mammals present in French Guiana, but dogs, capybaras, and opossums are the best candidate hosts for some of these filarioids. Although the detection of members of the Dipetalonema lineage in ticks of significant medical or veterinary interest is of concern, the risk of contracting a tick-borne filarial infection is still largely unknown. The pathogenicity of these filarioids, their epidemiology, developmental cycles, and mechanisms of transmission by South American tick species now require further study.

双足线虫谱系的丝状线虫是广泛存在的寄生虫,包括一些由蜱虫传播的物种。在这项研究中,我们对南美洲法属圭亚那的蜱虫进行了大规模的分子调查,以了解这个被茂密的热带森林覆盖的偏远地区蜱虫传播的丝虫病的整体多样性。在6属22种682只蜱中,卡詹钝肢蜱、长尾古纹蜱、罗米蒂蜱、露阴伊蚊和血感鼻头蜱有21只(3.1%)呈丝虫病染阳性。分子分型和系统发育分析鉴定所有丝状体均为双足线虫谱系的成员。虽然R. sanguineus sensu lato的丝状体是先前描述的物种,犬科蠕虫Cercopithifilaria bainae Almeida & Vicente, 1984,本研究中检测到的所有其他丝状体与已知的Cercopithifilaria, Cruorifilaria和Dipetalonema属的丝状体相关但不同。它们的脊椎动物宿主范围可能包括法属圭亚那存在的各种各样的哺乳动物,但狗、水豚和负鼠是这些丝虫病的最佳候选宿主。尽管在蜱虫中检测到双爪虫谱系成员具有重要的医学或兽医意义,但感染蜱虫传播的丝虫病的风险在很大程度上仍然未知。这些丝状虫的致病性、流行病学、发育周期和南美蜱类传播机制有待进一步研究。
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引用次数: 0
COMBAR - Combatting anthelmintic resistance in ruminants. COMBAR -对抗反刍动物的抗虫性。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023006
Johannes Charlier, Hervé Hoste, Smaro Sotiraki
Cattle, sheep and goats are parasitized by various helminth species, the most important being the gastrointestinal nematodes (GIN) and liver fl uke. These pathogens can cause severe disease and affect productivity in all classes of livestock and are worldwide amongst the most important production-limiting diseases of grazing ruminants [2]. Essentially, all herds/ fl ocks in a grass-based production system are affected and the major economic impact is due to sub-clinical infections causing reduced growth and milk/wool production. These parasitic worms can also cause severe distress and disease, affecting animal welfare, and through the impact on farm management, production and food security, indirectly human wellbeing.
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引用次数: 4
Therapeutic efficacy and safety of artesunate + amodiaquine and artemether + lumefantrine in treating uncomplicated Plasmodium falciparum malaria in children on the rainy south-east coast of Madagascar. 青蒿琥酯+阿莫地喹和蒿甲醚+氨苯曲明治疗多雨的马达加斯加东南海岸儿童无并发症恶性疟原虫疟疾的疗效和安全性
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023034
Judickaëlle Irinantenaina, Gwénaëlle Carn, Dina Ny Aina Liantsoa Randriamiarinjatovo, Aina Nirina Harimanana, Seheno Razanatsiorimalala, Nicolas Ralemary, Maurice Randriarison, Celestin Razafinjato, Raphael Hotahiene, Milijaona Randrianarivelojosia

Malaria is a major public health problem in Madagascar, particularly in coastal areas. We conducted a randomized, controlled, parallel-group study of artemisinin-based combination therapy (ACT) in Mananjary and Farafangana, two localities on the rainy south-east coast of Madagascar, from March to September 2018. The efficacy and safety of artesunate + amodiaquine (ASAQ) and artemether + lumefantrine (AL) were assessed according to the WHO protocol with a 28-day follow-up. Children aged 6 months to 14 years with uncomplicated Plasmodium falciparum malaria were randomized to receive ASAQ or AL for three days (1:1). 347/352 (98.5%) randomized patients reached the study endpoint on day 28. Crude adequate clinical and parasitological response (ACPR) rates were 100% (95% CI: 98.8-100%) in the ASAQ group and 96% (95% CI: 93.1-98.9%) in the AL group (per protocol population). However, the PCR-corrected ACPR rate was 97.7% (95% CI: 95.4-100%) in the AL group. Two cases of recrudescence and three of re-infection were observed. Mild and moderate adverse events, including gastrointestinal and/or nervous disorders, were reported in 11.9% (42/352) of patients. We found that ASAQ and AL were safe and efficacious for treating uncomplicated P. falciparum malaria. They may be used for treatment at health facilities and at the community level, and for mass drug administration campaigns.

疟疾是马达加斯加的一个主要公共卫生问题,特别是在沿海地区。2018年3月至9月,我们在马达加斯加多雨的东南海岸的Mananjary和Farafangana两个地区进行了一项随机、对照、平行组的青蒿素联合疗法(ACT)研究。通过28天的随访,根据WHO方案评估青蒿琥酯+阿莫地喹(ASAQ)和蒿甲醚+氨苯曲明(AL)的有效性和安全性。6个月至14岁无并发症恶性疟原虫疟疾患儿随机接受ASAQ或AL治疗3天(1:1)。347/352例(98.5%)随机患者在第28天到达研究终点。ASAQ组的临床和寄生虫反应(ACPR)率为100% (95% CI: 98.8-100%), AL组的ACPR率为96% (95% CI: 93.1-98.9%)。然而,AL组pcr校正的ACPR率为97.7% (95% CI: 95.4-100%)。复发2例,再感染3例。11.9%(42/352)的患者报告了轻度和中度不良事件,包括胃肠道和/或神经紊乱。我们发现ASAQ和AL治疗无并发症恶性疟原虫疟疾安全有效。它们可用于保健设施和社区一级的治疗,以及用于大规模药物管理运动。
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引用次数: 0
On the systematics of Phlebotomus betisi and two new related species from Laos with proposal of the new subgenus Lewisius. 老挝贝氏白蛉及两新近缘种的系统分类学研究及新亚属的建立。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023021
Khamsing Vongphayloth, Fano José Randrianambinintsoa, Khaithong Lakeomany, Nothasine Phommavanh, Nalia Mekarnia, Mohd Shahar Khadri, Matthieu L Kaltenbach, Antoine Huguenin, Jean-Philippe Martinet, Jérôme Depaquit

Phlebotomus betisi was described from Malaysia and classified after its description in the subgenus Larroussius. It was the only species to have a pharyngeal armature composed of dot-like teeth and an annealed spermatheca whose head is carried by a neck in females. Males were characterized by having a style bearing five spines and a simple paramere. The study of sandflies originating from a cave in Laos enabled us to discover and describe two sympatric species close to Ph. betisi Lewis & Wharton, 1963 and new for Science: Ph. breyi Vongphayloth & Depaquit n. sp., and Ph. sinxayarami Vongphayloth & Depaquit n. sp. They were characterized morphologically, morphometrically, geomorphometrically, molecularly, and proteomically (MALDI-TOF). All approaches converged to validate the individualization of these species whose morphological differential characters lay in the two genders by the observation of the interocular suture and by the length of the last two segments of the maxillary palps. In males, the length of the genital filaments discriminates these species. Females are distinguished by the length of the ducts of the spermathecae as well as by the narrow or enlarged shape of the neck bearing their head. Lastly, the particular position of the spines of the gonostyle coupled with molecular phylogeny led us to remove these three species from the subgenus Larroussius Nizulescu, 1931 and to classify them in a new subgenus: Lewisius Depaquit & Vongphayloth n. subg.

马来白蛉是马来西亚的一种昆虫,属Larroussius亚属。它是唯一一种有由点状牙齿组成的咽枢和一个退火的精子囊的物种,雌性的头部由颈部携带。男性的特征是花柱有五个刺和一个简单的参数。对源自老挝洞穴的白蛉的研究使我们发现并描述了两个与Ph. betisi Lewis & Wharton(1963)相近的同域物种:Ph. breyi Vongphayloth & Depaquit n. sp和Ph. sinxayarami Vongphayloth & Depaquit n. sp。它们在形态、形态计量学、地貌学、分子和蛋白质组学(MALDI-TOF)上进行了表征。通过观察眼间缝合线和上颌触须最后两段的长度,所有的方法都融合在一起,以验证这些物种的个体化,这些物种的形态差异特征在于两性。在雄性中,生殖器细丝的长度区分了这些物种。雌性的特点是精子管的长度以及颈部的狭窄或扩大的形状。最后,根据淋球菌棘的特殊位置和分子系统发育,我们将这三个种从Larroussius Nizulescu(1931)亚属中分离出来,并将它们分类为一个新的亚属:Lewisius Depaquit & Vongphayloth n. subg。
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引用次数: 0
Radiation dose-fractionation in adult Aedes aegypti mosquitoes. 埃及伊蚊成蚊的辐射剂量分级。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023005
Hanano Yamada, Hamidou Maïga, Carina Kraupa, Nanwintoum Séverin Bimbilé Somda, Wadaka Mamai, Thomas Wallner, Jeremy Bouyer

Balancing process efficiency and adult sterile male biological quality is one of the challenges in the success of the sterile insect technique (SIT) against insect pest populations. For the SIT against mosquitoes, many stress factors need to be taken into consideration when producing sterile males that require high biological quality to remain competitive once released in the field. Pressures of mass rearing, sex sorting, irradiation treatments, packing, transport and release including handling procedures for each step, add to the overall stress budget of the sterile male post-release. Optimizing the irradiation step to achieve maximum sterility while keeping off-target somatic damage to a minimum can significantly improve male mating competitiveness. It is therefore worth examining various protocols that have been found to be effective in other insect species, such as dose fractionation. A fully sterilizing dose of 70 Gy was administered to Aedes aegypti males as one acute dose or fractionated into either two equal doses of 35 Gy, or one low dose of 10 Gy followed by a second dose of 60 Gy. The two doses were separated by either 1- or 2-day intervals. Longevity, flight ability, and mating competitiveness tests were performed to identify beneficial effects of the various treatments. Positive effects of fractionating dose were seen in terms of male longevity and mating competitiveness. Although applying split doses generally improved male quality parameters, the benefits may not outweigh the added labor in SIT programmes for the management of mosquito vectors.

平衡过程效率和成虫不育雄虫的生物质量是昆虫不育技术成功防治害虫的挑战之一。对于抗蚊的SIT,在产生不育雄蚊时需要考虑许多应激因素,这些雄蚊需要高生物质量才能在野外释放后保持竞争力。大量饲养、性别分类、辐照处理、包装、运输和释放的压力,包括每个步骤的处理程序,增加了释放后不育雄性的总体压力预算。优化辐照步骤以达到最大的不育性,同时将脱靶体细胞损伤降至最低,可以显著提高雄性交配竞争力。因此,值得研究已被发现对其他昆虫物种有效的各种方案,例如剂量分馏。对埃及伊蚊雄性给予70戈瑞的完全绝育剂量,作为一次急性剂量,或分两次等量剂量(35戈瑞),或一次低剂量(10戈瑞),然后第二次剂量(60戈瑞)。两剂药间隔1天或2天。进行了寿命、飞行能力和交配竞争测试,以确定各种处理的有益效果。分离剂量对雄性寿命和交配竞争力均有积极影响。虽然采用分次剂量通常改善了雄性质量参数,但其好处可能不会超过为管理蚊子媒介而进行的SIT规划中增加的劳动。
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引用次数: 1
Nearctic Dactylogyrus species (Platyhelminthes, Monogenea) parasitizing cypriniform fishes in the context of morphology and phylogeny, with descriptions of seven new species. 寄生于鲤形鱼类的新北极Dactylogyrus种(platyhelmintes, Monogenea)的形态和系统发育,附7个新种的描述。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023032
Eva Řehulková, Mária Seifertová, Kateřina Francová, Andrea Šimková

Ribosomal DNA sequences are currently available for 32 morphologically recognized species of Dactylogyrus parasitizing Nearctic cypriniforms, but only 16 of them are assigned to nominal species. Herein, morphological data on 28 of the 32 species are provided, together with comments on their phylogenetic relationships in the context of the morphology of taxonomically important structures. Seven new species of Dactylogyrus are described from five species of leuciscids and one species of catostomids, as follows: D. aduncus n. sp. from Campostoma spadiceum, D. cloutmani n. sp. from Luxilus chrysocephalus isolepis, D. cornifrons n. sp. from Cyprinella venusta, D. fimbratus n. sp. from Rhinichthys cataractae, D. mcallisteri n. sp. from Semotilus atromaculatus, and D. chieni n. sp. and D. haneki n. sp. from Hypentelium nigricans. Four previously described species of Dactylogyrus, D. atromaculatus from S. atromaculatus, D. eos from C. neogaeus, D. parvicirrus from Notemigonus crysoleucas and D. perlus from L. c. isolepis, are redescribed and/or figured. As for the remaining 17 species, only the male copulatory organs (MCOs) taken from the respective hologenophores are illustrated. On the basis of phylogenetic analyses, two main clades of Nearctic Dactylogyrus were recognized and supported by the different morphology of the MCO. The first one included 22 strictly Nearctic species sharing the same MCO type with Dactylogyrus spp. parasitizing cyprinids likely of Asian origin. The second clade encompassed Dactylogyrus spp. with diverse MCO morphology and was placed in the sister position to Dactylogyrus spp. parasitizing European leuciscids and North-West African cyprinids.

目前已有32种寄生于新北极cypriniforms的Dactylogyrus的核糖体DNA序列,但其中只有16种属于名义种。本文提供了32种中28种的形态数据,并在分类上重要结构的形态背景下对它们的系统发育关系进行了评论。从5种leucisids和1种catostomids中发现7个Dactylogyrus新种:Campostoma spaduncus n. sp., Luxilus chrysocephalus cloutmani n., Cyprinella venusta ., Rhinichthys cataractae . fimbratus n., Semotilus atromaculatus D., chieni n.和haneki n.。本文重新描述了先前描述过的4种Dactylogyrus,即来自S. atromaculatus的D. atromaculatus,来自C. neogaeus的D. eos,来自Notemigonus crysoleucas的D. parvicirus和来自L. C. isolepis的D. perlus。至于其余17种,仅说明了从各自的全息孢子中提取的雄性交配器官(MCOs)。在系统发育分析的基础上,通过不同形态的MCO识别并支持了新北极趾类的两个主要分支。第一个包括22个严格的新北极物种,它们与可能来自亚洲的寄生鲤的Dactylogyrus属具有相同的MCO类型。第二支系包括具有不同MCO形态的长尾霉属,与寄生于欧洲幼蚊和西北非洲鲤的长尾霉属处于姐妹地位。
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引用次数: 0
Molecular phylogeny of Psychodopygina (Diptera, Psychodidae) supporting morphological systematics of this group of vectors of New World tegumentary leishmaniasis. 双翅目麻风蝇科麻风蝇的分子系统发育及对该群新发利什曼病媒介的形态学系统支持。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023018
Sonia Zapata, Eunice A B Galati, Jaime A Chaves, Patricio Artigas, Jean-Charles Gantier, Maria Dolores Bargues, Santiago Mas-Coma, Jérôme Depaquit

New World sandflies are vectors of leishmaniasis, bartonellosis, and some arboviruses. A classification based on 88 morphological characters was proposed 27 years ago when the New World phlebotomines were organized into two tribes Hertigiini and Phlebotomini. The latter was structured into four subtribes (Brumptomyiina, Sergentomyiina, Lutzomyiina, and Psychodopygina) and 20 genera. The subtribe Psychodopygina, including most of the American vectors of tegumentary Leishmania comprises seven genera from which no molecular work has been produced to support this classification. Here, we carried out a molecular phylogeny based on combined sequences (1,334 bp) of two genes: partial 28S rDNA and mtDNA cytochrome b from 47 taxa belonging to the Psychodopygina. The Bayesian phylogenetic reconstruction agreed with the classification based on morphological characters, supporting the monophyly of the genera Psychodopygus and Psathyromyia, whereas Nyssomyia and Trichophoromyia seemed to be paraphyletic. The paraphylies of the two latter groups were exclusively caused by the doubtful position of the species Ny. richardwardi. Our molecular analysis provides additional support to adopt the morphologic classification of Psychodopygina.

新大陆白蛉是利什曼病、巴尔通体病和一些虫媒病毒的带菌者。27年前,新世界白蛉被划分为Hertigiini和Phlebotomini两个部落,并提出了基于88个形态特征的分类。后者可分为4个亚族(Brumptomyiina、Sergentomyiina、Lutzomyiina和Psychodopygina)和20个属。包括大部分美洲被膜利什曼原虫病媒在内的精神涡虫亚族包括7个属,但尚未有分子研究支持这一分类。在此,我们基于47个属精神病属类群的28S rDNA和mtDNA细胞色素b两个基因的组合序列(1334 bp)进行了分子系统发育。贝叶斯系统发育重建结果与基于形态特征的分类一致,支持心理虫属和甲状虫属的单一性,而Nyssomyia和Trichophoromyia似乎是副属的。后两组的瘫痪完全是由物种Ny的可疑位置引起的。richardwardi。我们的分子分析为采用精神麻痹的形态学分类提供了额外的支持。
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引用次数: 0
Real-time multiplex PCR for human echinococcosis and differential diagnosis. 实时多重PCR检测人包虫病及鉴别诊断。
IF 2.9 2区 医学 Q1 Agricultural and Biological Sciences Pub Date : 2023-01-01 DOI: 10.1051/parasite/2023003
Jenny Knapp, Séverine Lallemand, Franck Monnien, Sophie Felix, Sandra Courquet, Gérald Umhang, Laurence Millon

Molecular identification of rare human infectious pathogens appears to be one of the most relevant current methods for rapid diagnosis and management of patients. PCR techniques, in particular real-time quantitative PCR, are best suited for the detection of DNA from the pathogens, even at low concentrations. Echinococcosis infections are due to helminths of the Echinococcus genus, with closely related species involved in parasitic lesions affecting animals and, accidentally, humans. We developed a multiplex qPCR (MLX qPCR) assay allowing for the detection of four Echinococcus species involved in Europe in alveolar echinococcosis (AE) and cystic echinococcosis (CE) (Echinococcus multilocularis, E. granulosus sensu stricto, E. ortleppi, and E. canadensis), based on short mitochondrial targets. A collection of 81 fresh and formalin-fixed paraffin-embedded tissues (FFPE) of AE and CE lesions was assembled. The qPCR assays were performed in triplex for Echinococcus spp. detection, associated with a qPCR inhibitor control. A duplex qPCR was also designed to enable diagnosis of two other dead-end helminthiases (cysticercosis (Taenia solium), and toxocariasis (Toxocara cati and T. canis)). The sensitivity of the qPCR was assessed and ranged from 1 to 5 × 10-4 ng/μL (seven PCR assays positive), corresponding to 37-42 cycles for quantifiable DNA. The specificity was 100% for all the targets. This multiplex qPCR, adapted to low amounts of DNA can be implemented in the laboratory for the rapid molecular diagnosis of Echinococcosis species.

罕见的人类传染性病原体的分子鉴定似乎是目前最相关的快速诊断和管理患者的方法之一。PCR技术,特别是实时定量PCR,最适合于检测来自病原体的DNA,即使在低浓度下也是如此。棘球绦虫感染是由棘球绦虫属的蠕虫引起的,与之密切相关的物种参与了影响动物和偶然影响人类的寄生虫病变。我们开发了一种基于短线粒体靶标的多重qPCR (MLX qPCR)方法,用于检测欧洲肺泡棘球蚴病(AE)和囊性棘球蚴病(CE)的四种棘球蚴(多房棘球蚴、狭义棘球蚴、北棘球蚴和加拿大棘球蚴)。收集了81例AE和CE病变的新鲜和福尔马林固定石蜡包埋组织(FFPE)。采用三联体qPCR检测棘球绦虫,并与qPCR抑制剂对照。设计了双工qPCR,用于诊断另外两种终端蠕虫病(猪带绦虫囊虫病)和弓形虫病(猫弓形虫和犬弓形虫)。qPCR的灵敏度范围为1 ~ 5 × 10-4 ng/μL(7次PCR检测为阳性),对应37 ~ 42个周期的可定量DNA。所有靶点的特异性均为100%。该多重qPCR适应于低DNA量,可在实验室实施棘球蚴病物种的快速分子诊断。
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引用次数: 6
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Parasite
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