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Helminth co-infections have no additive detrimental impact on milk yield and milk quality compared to mono-infections in German dairy cows 在德国奶牛中,与单一感染相比,螺旋体合并感染对奶牛的产奶量和牛奶质量没有额外的不利影响
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-19 DOI: 10.1186/s13071-024-06470-8
Katharina May, Anna Sophie Hecker, Sven König, Christina Strube
Infections with (tricho-)strongyles, Dictyocaulus viviparus or Fasciola hepatica have been shown to reduce milk production in dairy cows. However, the current published studies focused on one single helminth infection by neglecting helminth co-infections and their possible (additive) effects on host performance. Hence, for the first time, we investigated differences in the impact of patent helminth co-infections versus mono-infections on milk production parameters in individual cows. A total of 1583 dairy cows from 27 herds were included in this study. Faecal samples were examined in 2015 and 2021/2022 to determine the number of eggs/larvae per gram faeces for (tricho-)strongyles, D. viviparus, F. hepatica and rumen flukes. The cows were classified as non-infected, mono-infected and co-infected. Linear mixed models were applied to analyse the association between infection status (non-infected vs. mono-infected vs. co-infected) with milk yield, milk protein and milk fat content by including potential confounders. Infections with (tricho-)strongyles, D. viviparus, F. hepatica and rumen flukes were detected in 100%, 28.6%, 50.0% and 21.4% of herds, and 27.4%, 2.6%, 10.8% and 0.8% of faecal samples in 2015, while 100%, 0.0%, 86.7% and 60.0% of herds and 52.3%, 0.0%, 13.3% and 26.8% of faecal samples were positive in 2021/2022. Co-infections with two or more helminth taxa were detected in 74.4% of herds and 5.0% of faecal samples in 2015, and in 93.3% of herds and 21.7% of faecal samples in 2021/2022. The correlations between strongyle EPG, D. viviparus LPG and F. hepatica EPG were significantly positive in 2015. Significantly higher mean EPGs were identified in 2015 in faecal samples presenting co-infections with F. hepatica and one or two other helminth taxa than in faecal samples presenting F. hepatica mono-infections (P = 0.013). Although expected, the infection status (mono- or co-infected) had no significant impact on milk yield, milk protein and milk fat content in the linear mixed model analyses based on individual faecal examinations. Patent helminth co-infections had no additive detrimental impact on milk production parameters in the present study. This might be a result of presumably low worm burdens, but should be confirmed in future studies.
研究表明,奶牛感染(三)强线虫、Dictyocaulus viviparus或Fasciola hepatica会降低产奶量。然而,目前已发表的研究主要集中在一种单一的蠕虫感染上,而忽略了蠕虫合并感染及其对宿主表现可能产生的(叠加)影响。因此,我们首次研究了专利蠕虫共感染与单一感染对奶牛产奶量参数影响的差异。本研究共纳入了来自 27 个牧场的 1583 头奶牛。在 2015 年和 2021/2022 年对粪便样本进行了检测,以确定每克粪便中(三代)强直球虫、D. viviparus、F. hepatica 和瘤胃吸虫的虫卵/幼虫数量。奶牛被分为未感染、单一感染和混合感染。采用线性混合模型分析感染状态(未感染 vs. 单一感染 vs. 共同感染)与产奶量、乳蛋白和乳脂肪含量之间的关系,并将潜在的混杂因素考虑在内。2015年,100%、28.6%、50.0%和21.4%的牛群以及27.4%、2.6%、10.8%和0.8%的粪便样本检测到(三代)强直球虫、D. viviparus、F. hepatica和瘤胃吸虫感染,而2021/2022年,100%、0.0%、86.7%和60.0%的牛群以及52.3%、0.0%、13.3%和26.8%的粪便样本检测到阳性。2015年,74.4%的畜群和5.0%的粪便样本检测到两种或两种以上蠕虫类群的共感染,2021/2022年,93.3%的畜群和21.7%的粪便样本检测到共感染。2015 年,强蛲虫 EPG、D. viviparus LPG 和 F. hepatica EPG 之间的相关性显著为正。2015 年,在同时感染肝包虫和一种或两种其他蠕虫类群的粪便样本中,EPG 平均值明显高于只感染肝包虫的粪便样本(P = 0.013)。尽管在意料之中,但在基于个体粪便检查的线性混合模型分析中,感染状态(单一感染或混合感染)对产奶量、乳蛋白和乳脂肪含量没有显著影响。在本研究中,专利螺旋体合并感染对牛奶产量参数没有额外的不利影响。这可能是由于蠕虫负担较低所致,但应在今后的研究中加以证实。
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引用次数: 0
A novel cystatin in Psoroptes ovis var. cuniculi: molecular characterization, serodiagnostic potential, and its anti-inflammatory property on rabbit peripheral blood mononuclear cells 猫鼬变种中的一种新型胱抑素:分子特征、血清诊断潜力及其对兔外周血单核细胞的抗炎特性
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-19 DOI: 10.1186/s13071-024-06483-3
Xiaobin Gu, Fusheng Yang, Ce Wang, Jing Xu, Yane Li, Youping Liang, Je Fan, Fangyan Wu, Ran He, Hui Wang, Yue Xie
The ectoparasite Psoroptes ovis var. cuniculi causes substantial economic losses to the global rabbit industry. Currently, microscopy for identifying Psoroptes mite in skin scrapings, as the “diagnosis gold standard,” remains a challenge owing to its poor sensitivity in detecting low-level and/or early stage mite infestations. Additionally, Psoroptes infestations rapidly trigger cutaneous inflammation, thus the mites might produce some molecules to deal with the harmful effects of inflammation for their long-time survival on the host skin, but these molecules are still mostly unknown. To seek a sensitive diagnostic method and illuminate the new antiinflammatory molecules, we characterized a novel cystatin of P. ovis var. cuniculi (PsoCys) using bioinformatics and molecular biology methods. The results showed that PsoCys comprised the classical features of the type II cystatin superfamily including an N-terminal glycine residue, a central QXVXG motif, and a C-terminal LW motif. In mixed stages of mites, the transcription level of PsoCys was significantly higher in “fed” mites than in “starved” mites (P < 0.001), and among the different life-cycle stages of “fed” mites, the expression of PsoCys was higher in adult males than in larva, nymph, and adult females (P < 0.001). The established indirect ELISA based on recombinant PsoCys (rPsoCys-iELISA) presented 95.4% sensitivity and 95.7% specificity. The area under the receiver operating characteristic curve (AUC) for this method was 0.991, indicating its excellent diagnostic performance. Moreover, rPsoCys-iELISA had advantages over microscopy for detecting low-level and/or early stage mite infestations (90% versus 40% in artificial infestation cases at 3 weeks post-infestation; 61.9% versus 22.6% in clinical cases). In addition, rPsoCys could inhibit the activity of papain and cathepsin B in vitro, and significantly suppressed mRNA levels of toll-like receptors (TLR 1, 2, 4, and 6) and downstream molecules (NF-κB, p38, MyD88, IL-10, and IFN-γ) in LPS-stimulated rabbit PBMCs, indicating its anti-inflammatory property. Our findings indicated that PsoCys was a novel type II cystatin of Psoroptes mites, and it served as a potential serological diagnostic antigen for detecting low-level and/or early stage mite infestations, as well as a novel anti-inflammatory molecule of Psoroptes mites.
外寄生虫 Psoroptes ovis var. cuniculi 给全球养兔业造成了巨大的经济损失。目前,显微镜作为 "诊断金标准",在皮肤刮片中识别Psoroptes螨仍然是一项挑战,因为它在检测低水平和/或早期螨虫感染方面灵敏度较低。此外,螨虫感染会迅速引发皮肤炎症,因此螨虫可能会产生一些分子来应对炎症的有害影响,以便在宿主皮肤上长期存活,但这些分子大多仍不为人所知。为了寻求一种灵敏的诊断方法并揭示新的抗炎分子,我们利用生物信息学和分子生物学方法对一种新型的猫螨胱抑素(PsoCys)进行了表征。结果表明,PsoCys 具有 II 型胱抑素超家族的经典特征,包括一个 N 端甘氨酸残基、一个中央 QXVXG 矩阵和一个 C 端 LW 矩阵。在混合阶段的螨虫中,"喂养 "螨虫的 PsoCys 转录水平明显高于 "饥饿 "螨虫(P < 0.001),而且在 "喂养 "螨虫的不同生命周期阶段中,成年雄螨的 PsoCys 表达高于幼虫、若螨和成年雌螨(P < 0.001)。基于重组 PsoCys 的间接 ELISA(rPsoCys-iELISA)灵敏度为 95.4%,特异性为 95.7%。该方法的接收者操作特征曲线下面积(AUC)为 0.991,表明其诊断性能极佳。此外,与显微镜相比,rPsoCys-iELISA 在检测低水平和/或早期螨虫感染方面更具优势(在感染后 3 周的人工感染病例中,检测率为 90% 对 40%;在临床病例中,检测率为 61.9% 对 22.6%)。此外,rPsoCys 还能在体外抑制木瓜蛋白酶和 cathepsin B 的活性,并能显著抑制 LPS 刺激的兔 PBMC 中的收费样受体(TLR 1、2、4 和 6)和下游分子(NF-κB、p38、MyD88、IL-10 和 IFN-γ)的 mRNA 水平,这表明它具有抗炎特性。我们的研究结果表明,PsoCys 是一种新型的 Psoroptes 螨虫 II 型胱抑素,可作为一种潜在的血清学诊断抗原用于检测低水平和/或早期阶段的螨虫感染,同时也是一种新型的 Psoroptes 螨虫抗炎分子。
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引用次数: 0
Seasonal dynamics, Leishmania diversity, and nanopore-based metabarcoding of blood meal origins in Culicoides spp. in the newly emerging focus of leishmaniasis in Northern Thailand. 泰国北部新出现的利什曼病病灶中库利科氏虫的季节动态、利什曼原虫多样性以及基于纳米孔的血粉来源代谢编码。
IF 3 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-19 DOI: 10.1186/s13071-024-06487-z
Chulaluk Promrangsee, Sira Sriswasdi, Sakone Sunantaraporn, Chatuthanai Savigamin, Thanapat Pataradool, Chatchapon Sricharoensuk, Rungfar Boonserm, Rinnara Ampol, Pitchayaporn Pruenglampoo, Mathirut Mungthin, Jonas Schmidt-Chanasit, Padet Siriyasatien, Kanok Preativatanyou
<p><strong>Background: </strong>Clinical cases of leishmaniasis caused by Leishmania (Mundinia) parasites have been increasingly reported in Southeast Asia, particularly Thailand. Recent evidence has shown that Leishmania (Mundinia) parasites successfully developed into infective metacyclic promastigotes in Culicoides biting midges, strongly supporting their putative role in disease transmission. However, Culicoides diversity, host preference, and Leishmania prevalence in endemic areas remain largely unknown.</p><p><strong>Methods: </strong>We investigated the seasonal dynamics, infection prevalence, and blood meal identification of Culicoides collected from the emerging focus of visceral leishmaniasis in Lampang Province, Northern Thailand, during 2021-2023. Midge samples were molecularly screened for Leishmania using SSU rRNA-qPCR and ITS1-PCR, followed by Sanger plasmid sequencing, and parasite haplotype diversity was analyzed. Host blood meal origins were comparatively identified using host-specific Cytb-PCRs and a nanopore-based metabarcoding approach.</p><p><strong>Results: </strong>A total of 501 parous and gravid females and 46 blood-engorged ones belonging to at least 17 species of five subgenera (Remmia, Trithecoides, Avaritia, Hoffmania, and Meijerehelea) and two species groups (Shortti and Calvipalpis) were collected with temporal differences in abundance. Leishmania was detected by SSU rRNA-qPCR in 31 samples of at least 11 midge species, consisting of Culicoides oxystoma, C. guttifer, C. orientalis, C. mahasarakhamense, C (Trithecoides) spp., C. innoxius, C. shortti, C. arakawae, C. sumatrae, C. actoni, and C. fulvus, with the overall infection prevalence of 5.7%. The latter six species represent the new records as putative leishmaniasis vectors in Northern Thailand. The ITS1-PCR and plasmid sequencing revealed that Leishmania martiniquensis was predominantly identified in all qPCR-positive species, whereas L. orientalis was identified only in three C. oxystoma samples. The most dominant haplotype of L. martiniquensis in Thailand was genetically intermixed with those from other geographical regions, confirming its globalization. Neutrality test statistics were also significantly negative on regional and country-wide scales, suggesting rapid population expansion or selective sweeps. Nanopore-based blood meal analysis revealed that most Culicoides species are mammalophilic, with peridomestic and wild mammals (cow, pig, deer, and goat-like species) and humans as hosts, while C. guttifer and C. mahasarakhamense fed preferentially on chickens.</p><p><strong>Conclusions: </strong>This study revealed seasonal dynamics and sympatric circulation of L. martiniquensis and L. orientalis in different species of Culicoides. Evidence of human blood feeding was also demonstrated, implicating Culicoides as putative vectors of human leishmaniasis in endemic areas. Further research is therefore urgently needed to develop vector control strategies and
背景:在东南亚,尤其是泰国,利什曼病(Mundinia)寄生虫引起的利什曼病临床病例报告越来越多。最近的证据表明,利什曼原虫(Mundinia)寄生虫成功地在咬蠓 Culicoides 中发育成具有感染性的中生原体,有力地支持了它们在疾病传播中的假定作用。然而,Culicoides 的多样性、宿主偏好以及利什曼病流行地区的利什曼病发病率在很大程度上仍是未知数:方法:我们调查了 2021-2023 年期间从泰国北部南邦省新出现的内脏利什曼病重点地区采集的蠓类的季节动态、感染率和血餐鉴定。使用 SSU rRNA-qPCR 和 ITS1-PCR 对蠓样本进行利什曼原虫分子筛选,然后进行 Sanger 质粒测序,并分析寄生虫单倍型多样性。使用宿主特异性 Cytb-PCR 和基于纳米孔的代谢编码方法比较鉴定了宿主血餐来源:结果:共收集到 501 只雌虫和 46 只血食性雌虫,它们分别属于 5 个亚属(Remmia、Trithecoides、Avaritia、Hoffmania 和 Meijerehelea)和 2 个种群(Shortti 和 Calvipalpis)中的至少 17 个种,且数量存在时间差异。通过 SSU rRNA-qPCR,在至少 11 种蠓类的 31 份样本中检测到了利什曼原虫,包括 Culicoides oxystoma、C. guttifer、C. orientalis、C. mahasarakhamense、C (Trithecoides) spp.、C. innoxius、C. shortti、C. arakawae、C. sumatrae、C. actoni 和 C. fulvus,总体感染率为 5.7%。后六种是泰国北部利什曼病病媒的新记录。ITS1-PCR 和质粒测序结果显示,在所有 qPCR 阳性的物种中主要发现了马氏利什曼病,而仅在三个 C. oxystoma 样本中发现了东方利什曼病。泰国最主要的 L. martiniquensis 单倍型与其他地理区域的单倍型在基因上混杂在一起,证实了它的全球化。在地区和国家范围内,中性检验统计量也呈显著负值,这表明种群正在迅速扩大或进行选择性扫荡。基于纳米孔的血粉分析表明,大多数Culicoides物种都嗜哺乳动物,以家畜和野生哺乳动物(牛、猪、鹿和山羊类物种)以及人类为宿主,而C. guttifer和C. mahasarakhamense则以鸡为食:这项研究揭示了马氏栉水母和东方栉水母在不同物种中的季节性动态和同域循环。此外,还发现了以人类血液为食的证据,这表明在利什曼病流行地区,Culicoides可能是人类利什曼病的传播媒介。因此,迫切需要开展进一步研究,以制定病媒控制战略,并评估其储藏宿主的感染状况,从而有效地将疾病传播降至最低。
{"title":"Seasonal dynamics, Leishmania diversity, and nanopore-based metabarcoding of blood meal origins in Culicoides spp. in the newly emerging focus of leishmaniasis in Northern Thailand.","authors":"Chulaluk Promrangsee, Sira Sriswasdi, Sakone Sunantaraporn, Chatuthanai Savigamin, Thanapat Pataradool, Chatchapon Sricharoensuk, Rungfar Boonserm, Rinnara Ampol, Pitchayaporn Pruenglampoo, Mathirut Mungthin, Jonas Schmidt-Chanasit, Padet Siriyasatien, Kanok Preativatanyou","doi":"10.1186/s13071-024-06487-z","DOIUrl":"https://doi.org/10.1186/s13071-024-06487-z","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Background: &lt;/strong&gt;Clinical cases of leishmaniasis caused by Leishmania (Mundinia) parasites have been increasingly reported in Southeast Asia, particularly Thailand. Recent evidence has shown that Leishmania (Mundinia) parasites successfully developed into infective metacyclic promastigotes in Culicoides biting midges, strongly supporting their putative role in disease transmission. However, Culicoides diversity, host preference, and Leishmania prevalence in endemic areas remain largely unknown.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;We investigated the seasonal dynamics, infection prevalence, and blood meal identification of Culicoides collected from the emerging focus of visceral leishmaniasis in Lampang Province, Northern Thailand, during 2021-2023. Midge samples were molecularly screened for Leishmania using SSU rRNA-qPCR and ITS1-PCR, followed by Sanger plasmid sequencing, and parasite haplotype diversity was analyzed. Host blood meal origins were comparatively identified using host-specific Cytb-PCRs and a nanopore-based metabarcoding approach.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;A total of 501 parous and gravid females and 46 blood-engorged ones belonging to at least 17 species of five subgenera (Remmia, Trithecoides, Avaritia, Hoffmania, and Meijerehelea) and two species groups (Shortti and Calvipalpis) were collected with temporal differences in abundance. Leishmania was detected by SSU rRNA-qPCR in 31 samples of at least 11 midge species, consisting of Culicoides oxystoma, C. guttifer, C. orientalis, C. mahasarakhamense, C (Trithecoides) spp., C. innoxius, C. shortti, C. arakawae, C. sumatrae, C. actoni, and C. fulvus, with the overall infection prevalence of 5.7%. The latter six species represent the new records as putative leishmaniasis vectors in Northern Thailand. The ITS1-PCR and plasmid sequencing revealed that Leishmania martiniquensis was predominantly identified in all qPCR-positive species, whereas L. orientalis was identified only in three C. oxystoma samples. The most dominant haplotype of L. martiniquensis in Thailand was genetically intermixed with those from other geographical regions, confirming its globalization. Neutrality test statistics were also significantly negative on regional and country-wide scales, suggesting rapid population expansion or selective sweeps. Nanopore-based blood meal analysis revealed that most Culicoides species are mammalophilic, with peridomestic and wild mammals (cow, pig, deer, and goat-like species) and humans as hosts, while C. guttifer and C. mahasarakhamense fed preferentially on chickens.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;This study revealed seasonal dynamics and sympatric circulation of L. martiniquensis and L. orientalis in different species of Culicoides. Evidence of human blood feeding was also demonstrated, implicating Culicoides as putative vectors of human leishmaniasis in endemic areas. Further research is therefore urgently needed to develop vector control strategies and ","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"17 1","pages":"400"},"PeriodicalIF":3.0,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11414243/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142292836","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
Whole genome sequence analysis of population structure and insecticide resistance markers in Anopheles melas from the Bijagós Archipelago, Guinea-Bissau 几内亚比绍比热戈斯群岛梅拉按蚊种群结构和抗杀虫剂标记的全基因组序列分析
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-18 DOI: 10.1186/s13071-024-06476-2
Sophie Moss, Elizabeth Pretorius, Sainey Ceesay, Eunice Teixeira da Silva, Harry Hutchins, Mamadou Ousmane Ndiath, Holly Acford-Palmer, Emma L. Collins, Matthew Higgins, Jody Phelan, Robert T. Jones, Hristina Vasileva, Amabelia Rodrigues, Sanjeev Krishna, Taane G. Clark, Anna Last, Susana Campino
Anopheles melas is an understudied malaria vector with a potential role in malaria transmission on the Bijagós Archipelago of Guinea-Bissau. This study presents the first whole-genome sequencing and population genetic analysis for this species from the Bijagós. To our knowledge, this also represents the largest population genetic analysis using WGS data from non-pooled An. melas mosquitoes. WGS was conducted for 30 individual An. melas collected during the peak malaria transmission season in 2019 from six different islands on the Bijagós Archipelago. Bioinformatics tools were used to investigate the population structure and prevalence of insecticide resistance markers in this mosquito population. Insecticide resistance mutations associated with pyrethroid resistance in Anopheles gambiae s.s. from the Bijagós were absent in the An. melas population, and no signatures of selective sweeps were identified in insecticide resistance-associated genes. Analysis of structural variants identified a large duplication encompassing the cytochrome-P450 gene cyp9k1. Phylogenetic analysis using publicly available mitochondrial genomes indicated that An. melas from the Bijagós split into two phylogenetic groups because of differentiation on the mitochondrial genome attributed to the cytochrome C oxidase subunits COX I and COX II and the NADH dehydrogenase subunits 1, 4, 4L and 5. This study identified an absence of insecticide-resistant SNPs common to An. gambiae in the An. melas population, but did identify structural variation over insecticide resistance-associated genes. Furthermore, this study presents novel insights into the population structure of this malaria vector using WGS analysis. Additional studies are required to further understand the role of this vector in malaria transmission.
梅拉按蚊是一种未被充分研究的疟疾病媒,可能在几内亚比绍比热戈斯群岛的疟疾传播中扮演重要角色。本研究首次对比热戈斯群岛的这一物种进行了全基因组测序和种群遗传分析。据我们所知,这也是使用 WGS 数据对非汇集的瓜氏疟蚊进行的最大规模的种群遗传分析。在 2019 年疟疾传播高峰期,我们从比热戈斯群岛的六个不同岛屿上采集了 30 只 An. melas 个体,并对其进行了 WGS 分析。利用生物信息学工具研究了该蚊子种群的种群结构和杀虫剂抗药性标记的流行情况。在比热戈斯群岛的冈比亚按蚊种群中,与拟除虫菊酯抗性相关的杀虫剂抗性突变在梅拉斯按蚊种群中并不存在,在杀虫剂抗性相关基因中也未发现选择性扫描的特征。对结构变异的分析发现了细胞色素-P450基因cyp9k1的大量重复。利用公开的线粒体基因组进行的系统进化分析表明,由于细胞色素 C 氧化酶亚基 COX I 和 COX II 以及 NADH 脱氢酶亚基 1、4、4L 和 5 在线粒体基因组上的分化,来自比热戈斯的黑角蚁分成了两个系统进化组。这项研究发现梅拉斯鳗种群中没有冈比亚鳗常见的抗杀虫剂 SNPs,但确实发现了抗杀虫剂相关基因的结构变异。此外,本研究还利用 WGS 分析对这种疟疾病媒的种群结构提出了新的见解。要进一步了解这种病媒在疟疾传播中的作用,还需要进行更多的研究。
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引用次数: 0
Ongoing transmission of onchocerciasis in the Pru District of Ghana after two decades of mass drug administration with ivermectin and comparative identification of members of the Simulium damnosum complex using cytological and morphological techniques 使用伊维菌素大规模施药二十年后盘尾丝虫病在加纳普鲁地区的持续传播,以及使用细胞学和形态学技术对蚋复合体成员进行比较鉴定
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-18 DOI: 10.1186/s13071-024-06333-2
Friday Maduka Chikezie, Francis Balunnaa Dhari Veriegh, Samuel Armoo, Daniel Adjei Boakye, Mark Taylor, Mike Yaw Osei-Atweneboana
Human onchocerciasis remains a public health problem in Ghana. Mass drug administration (MDA) with ivermectin (IVM) has reduced disease morbidity and prevalence, but the transmission of onchocerciasis remains ongoing in several endemic foci. We investigated parasite transmission in some endemic communities in Ghana that had received > 18 rounds of annual MDA with IVM and determined the species composition of black fly (Simulium damnosum) vectors in these areas. Adult female black flies were collected using human landing catches and identified as either forest or savanna species using morpho-taxonomic keys. The adult flies underwent dissection to determine their parity and detect any O. volvulus larvae, followed by the calculation of entomological indices. Simulium damnosum s.l. larvae were collected and preserved in freshly prepared Carnoy’s fixative and were later used for cytotaxonomic studies. A total of 9,983 adult flies were caught: 6,569 and 3,414 in the rainy and dry seasons respectively. Black fly biting activities over the study period showed bimodal or trimodal patterns. The highest monthly biting rate (MBR) of 10,578.75 bites/person/month was recorded in July in Beposo, while the highest monthly transmission potential of 100.69 infective bites/person/month was recorded in Asubende in August. Morphological analysis of 2,032 flies showed that 99.8% (2,028) of the flies were savanna species, with only 4 (0.2%) adult flies being of the forest species. Cytogenetic studies on 114 black fly larvae revealed three cytospecies (Simulium damnosum s.s., S. sirbanum and S. sanctipauli) in the study area. The present studies confirmed an ongoing transmission of onchocerciasis in the study communities except Abua-1. It also provides further information on biting behaviors and onchocerciasis transmission indices in the study communities. Further, our data confirmed the savanna species (S. damnosum s.s. and S. sirbanum) of the S. damnosum s.l. to be the major vectors of onchocerciasis in the study areas, with only an occasional influx of forest cytotypes.
人类盘尾丝虫病仍然是加纳的一个公共卫生问题。使用伊维菌素(IVM)进行大规模药物治疗(MDA)降低了疾病的发病率和流行率,但盘尾丝虫病仍在几个流行病区持续传播。我们调查了加纳一些流行社区的寄生虫传播情况,这些社区每年接受的伊维菌素MDA超过18轮,并确定了这些地区黑蝇(Simulium damnosum)病媒的物种组成。利用人体着陆捕获物收集成年雌性黑蝇,并利用形态分类学钥匙将其鉴定为森林或热带草原物种。对成蝇进行解剖,以确定其奇偶性并检测是否存在卷叶黑蝇幼虫,然后计算昆虫学指数。Simulium damnosum s.l.幼虫被收集并保存在新鲜的卡诺氏固定液中,随后用于细胞分类学研究。总共捕获了 9,983 只成蝇:雨季为 6,569 只,旱季为 3,414 只。研究期间的黑蝇叮咬活动呈现出双峰或三峰模式。7 月份贝波索的月叮咬率(MBR)最高,达到 10,578.75 次/人/月,而 8 月份阿苏本德的月传播潜力最高,达到 100.69 次/人/月。对 2,032 只苍蝇的形态分析表明,99.8%(2,028 只)的苍蝇属于热带草原物种,只有 4 只(0.2%)成蝇属于森林物种。对 114 只黑蝇幼虫进行的细胞遗传学研究显示,研究地区有三个细胞种(Simulium damnosum s.s.、S. sirbanum 和 S. sanctipauli)。本研究证实,除 Abua-1 社区外,盘尾丝虫病在其他研究社区仍在传播。本研究还提供了有关研究社区的叮咬行为和盘尾丝虫病传播指数的进一步信息。此外,我们的数据证实,在研究地区,S. damnosum s.l.的热带草原种(S. damnosum s.s.和S. sirbanum)是盘尾丝虫病的主要传播媒介,森林细胞型只是偶尔出现。
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引用次数: 0
Exploring Trypanosoma cruzi transmission dynamics in an acute Chagas disease outbreak using next-generation sequencing 利用下一代测序技术探索南美锥虫病急性爆发期的传播动态
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-18 DOI: 10.1186/s13071-024-06445-9
Lissa Cruz-Saavedra, Carlos Ospina, Stivenn A. Gutiérrez, Jeiczon Jaimes-Dueñez, Omar Cantillo-Barraza, Carolina Hernández, Francisco Álvarez, María Blanco, Bernardo Leal, Lida Martínez, Manuel Medina, Mabel Medina, Silvia Valdivieso, Lauren Natalia Ramirez Celis, Luz H. Patiño, Juan David Ramírez
Chagas disease (CD), caused by Trypanosoma cruzi, poses a major global public health challenge. Although vector-borne transmission is the primary mode of infection, oral transmission is increasingly concerning. This study utilized long-amplicon-based sequencing (long-ABS), focusing on the 18S rRNA gene, to explore T. cruzi’s genetic diversity and transmission dynamics during an acute CD outbreak in Colombia, an area without domestic infestation. Analyzing samples from five patients and five T. cruzi-positive marsupial samples, we identified coinfections between T. cruzi and Trypanosoma rangeli, mixed T. cruzi DTUs, suggesting possible links between human and marsupial T. cruzi infections. Coexistence of TcI, TcIV and T. rangeli suggests marsupial secretions as the possible source of T. cruzi transmission. Our investigation revealed diversity loss in DTUs TcIV and T. rangeli in humans after infection and in marsupial samples after culture. These findings provide significant insights into T. cruzi dynamics, crucial for implementing control and prevention strategies.
由克鲁兹锥虫引起的恰加斯病(CD)是全球公共卫生面临的一大挑战。虽然病媒传播是主要的感染模式,但口腔传播也越来越令人担忧。这项研究利用基于长扩增子的测序(long-ABS)技术,以18S rRNA基因为重点,探讨了在哥伦比亚(一个没有国内侵扰的地区)爆发急性CD疫情期间,克鲁兹锥虫的遗传多样性和传播动态。通过分析五例患者样本和五例T. cruzi阳性有袋动物样本,我们发现了T. cruzi和Trypanosoma rangeli、混合T. cruzi DTUs之间的共感染,这表明人类和有袋动物T. cruzi感染之间可能存在联系。TcI、TcIV和T. rangeli的共存表明有袋类动物的分泌物可能是T. cruzi的传播源。我们的调查发现,在人类感染后和有袋动物样本培养后,DTUs TcIV 和 T. rangeli 的多样性丧失。这些发现为我们提供了有关 T. cruzi 动态的重要信息,对于实施控制和预防策略至关重要。
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引用次数: 0
Anisakis extracellular vesicles elicit immunomodulatory and potentially tumorigenic outcomes on human intestinal organoids 疟原虫胞外囊泡对人体肠道器官组织产生免疫调节和潜在致瘤作用
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-17 DOI: 10.1186/s13071-024-06471-7
Ilaria Bellini, Daniela Scribano, Cecilia Ambrosi, Claudia Chiovoloni, Silvia Rondón, Annamaria Pronio, Anna Teresa Palamara, Agostina Pietrantoni, Anna Kashkanova, Vahid Sandoghdar, Stefano D’Amelio, Serena Cavallero
Anisakis spp. are zoonotic nematodes causing mild to severe acute and chronic gastrointestinal infections. Chronic anisakiasis can lead to erosive mucosal ulcers, granulomas and inflammation, potential tumorigenic triggers. How Anisakis exerts its pathogenic potential through extracellular vesicles (EVs) and whether third-stage infective larvae may favor a tumorigenic microenvironment remain unclear. Here, we investigated the parasite's tumorigenic and immunomodulatory capabilities using comparative transcriptomics, qRT-PCR and protein analysis with multiplex ELISA on human intestinal organoids exposed to Anisakis EVs. Moreover, EVs were characterized in terms of shape, size and concentration using classic TEM, SEM and NTA analyses and advanced interferometric NTA. Anisakis EVs showed classic shape features and a median average diameter of around 100 nm, according to NTA and iNTA. Moreover, a refractive index of 5–20% of non-water content suggested their effective biological cargo. After treatment of human intestinal organoids with Anisakis EVs, an overall parasitic strategy based on mitigation of the immune and inflammatory response was observed. Anisakis EVs impacted gene expression of main cytokines, cell cycle regulation and protein products. Seven key genes related to cell cycle regulation and apoptosis were differentially expressed in organoids exposed to EVs. In particular, the downregulation of EPHB2 and LEFTY1 and upregulation of NUPR1 genes known to be associated with colorectal cancer were observed, suggesting their involvement in tumorigenic microenvironment. A statistically significant reduction in specific mediators of inflammation and cell-cycle regulation from the polarized epithelium as IL-33R, CD40 and CEACAM1 from the apical chambers and IL-1B, GM-CSF, IL-15 and IL-23 from both chambers were observed. The results here obtained unravel intestinal epithelium response to Anisakis EVs, impacting host’s anthelminthic strategies and revealing for the first time to our knowledge the host-parasite interactions in the niche environment of an emerging accidental zoonosis. Use of an innovative EV characterization approach may also be useful for study of other helminth EVs, since the knowledge in this field is very limited.
疟原虫是人畜共患的线虫,可引起轻度到重度的急性和慢性胃肠道感染。慢性恙虫病可导致侵蚀性粘膜溃疡、肉芽肿和炎症,是潜在的肿瘤诱因。目前尚不清楚恙虫病如何通过胞外囊泡(EVs)发挥其致病潜能,也不清楚第三阶段的感染性幼虫是否有利于肿瘤发生的微环境。在这里,我们使用比较转录组学、qRT-PCR 和蛋白分析(多重 ELISA)研究了暴露于 Anisakis EVs 的人体肠道有机体中的寄生虫的致癌和免疫调节能力。此外,还利用传统的 TEM、SEM 和 NTA 分析以及先进的干涉 NTA 分析,对 EVs 的形状、大小和浓度进行了表征。根据 NTA 和 iNTA,Anisakis EVs 表现出典型的形状特征,中位平均直径约为 100 纳米。此外,非水含量为 5-20% 的折射率表明它们是有效的生物载体。用疟原虫EVs处理人体肠道有机体后,观察到一种基于减轻免疫和炎症反应的整体寄生策略。Anisakis EVs 影响了主要细胞因子、细胞周期调控和蛋白质产物的基因表达。与细胞周期调控和细胞凋亡有关的七个关键基因在暴露于EVs的器官组织中表达不同。特别是观察到 EPHB2 和 LEFTY1 基因下调,而已知与结直肠癌相关的 NUPR1 基因上调,这表明它们参与了肿瘤微环境。据统计,极化上皮细胞的特定炎症介质和细胞周期调控介质明显减少,如顶端腔的 IL-33R、CD40 和 CEACAM1 以及两个腔的 IL-1B、GM-CSF、IL-15 和 IL-23。本研究的结果揭示了肠上皮细胞对疟原虫 EVs 的反应,影响了宿主的抗蠕虫策略,并首次揭示了宿主与寄生虫在新出现的意外人畜共患病的生态位环境中的相互作用。由于该领域的知识非常有限,因此使用创新的EV表征方法可能也有助于研究其他蠕虫EV。
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引用次数: 0
Evaluation of a long-lasting microbial larvicide against Culex quinquefasciatus and Aedes aegypti under laboratory and a semi-field trial 在实验室和半田间试验中评估针对库蚊和埃及伊蚊的长效微生物杀幼虫剂
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-14 DOI: 10.1186/s13071-024-06465-5
Hyago Luiz Rique, Heverly Suzany Gouveia Menezes, Maria Alice Varjal Melo-Santos, Maria Helena Neves Lobo Silva-Filha
Microbial larvicides containing both LysiniBacillus sphaericus and Bacillus thuringiensis svar. israelensis (Bti) insecticidal crystals can display advantages for mosquito control. This includes a broader action against larvae that are refractory to the Binary (Bin) toxin from L. sphaericus, as Bin-resistant Culex quinquefasciatus and Aedes aegypti naturally refractory larvae, which often co-habit urban areas of endemic countries for arboviruses. Our principal goal was to assess the toxicity of a combined L. sphaericus/Bti larvicide (Vectomax FG™) to Cx. quinquefasciatus (susceptible CqS and Bin-resistant CqR) and Ae. aegypti (Rocke) and to determine its persistence in the breeding sites with those larvae. The toxicity of a combined L. sphaericus/Bti product (VectoMax FG™) to larvae was performed using bioassays, and persistence was evaluated in simulate field trials carried out under the shade, testing two label concentrations during 12 weeks. A laboratory strain SREC, established with CqS and CqR larvae, was kept during four generations to evaluate the ability of the L. sphaericus/Bti to eliminate resistant larvae. The L. sphaericus/Bti showed toxicity (mg/L) to larvae from all strains with a decreasing pattern for CqS (LC50 = 0.006, LC90 = 0.030), CqR (LC50 = 0.009, LC90 = 0.069), and Rocke (LC50 = 0.042, LC90 = 0.086). In a simulated field trial, the larvicide showed a persistence of 6 weeks and 8 weeks, controlling larvae from all strains in containers with 100 L of water, using 2 g or 4 g per container (100 L), respectively. The treatment of SREC larvae with L. sphaericus/Bti showed its capacity to eliminate the Bin-resistant individuals using suitable concentrations to target those larvae. Our results showed the high efficacy and persistence of the L. sphaericus/Bti larvicide to control Cx. quinquefasciatus and Ae. aegypti that might cohabit breeding sites. These findings demonstrated that such larvicides can be an effective tool for controlling those species in urban areas with a low potential for selecting resistance.
同时含有 LysiniBacillus sphaericus 和 Bacillus thuringiensis svar. israelensis(Bti)杀虫晶体的微生物杀幼虫剂在控制蚊虫方面具有优势。这包括更广泛地作用于对唾液腺芽孢杆菌二元毒素(Bin)耐药的幼虫,如对Bin耐药的库蚊和埃及伊蚊天然耐药幼虫,它们经常共同生活在虫媒病毒流行国家的城市地区。我们的主要目标是评估 L. sphaericus/Bti 组合杀幼虫剂(Vectomax FG™)对五步蛇(易感 CqS 和 Bin 抗性 CqR)和埃及伊蚊(Rocke)的毒性,并确定其在这些幼虫繁殖地的持久性。使用生物测定法对 L. sphaericus/Bti 组合产品(VectoMax FG™)对幼虫的毒性进行了测定,并在 12 周内对两种标签浓度的产品进行了测试,在阴凉处进行的模拟现场试验中对其持久性进行了评估。用 CqS 和 CqR 幼虫建立的实验室菌株 SREC 经过四代饲养,以评估 L. sphaericus/Bti 清除抗性幼虫的能力。L. sphaericus/Bti 对所有菌株的幼虫都有毒性(毫克/升),对 CqS(LC50 = 0.006,LC90 = 0.030)、CqR(LC50 = 0.009,LC90 = 0.069)和 Rocke(LC50 = 0.042,LC90 = 0.086)的毒性呈下降趋势。在模拟田间试验中,杀幼虫剂的持效期分别为 6 周和 8 周,在装有 100 升水的容器中,每个容器(100 升)分别使用 2 克或 4 克,可控制所有菌株的幼虫。用 L. sphaericus/Bti 处理 SREC 幼虫的结果表明,用适当的浓度来消灭对 Bin 产生抗药性的幼虫个体是可行的。我们的研究结果表明,L. sphaericus/Bti杀幼虫剂对控制可能共居繁殖地的五步蛇和埃及伊蚊具有很高的效力和持久性。这些研究结果表明,这种杀幼虫剂可以成为控制城市地区这些物种的有效工具,而且产生抗药性的可能性较低。
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引用次数: 0
Identification and function characterization of NcAP2XII-4 in Neospora caninum 犬新孢子虫中 NcAP2XII-4 的鉴定和功能表征
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-14 DOI: 10.1186/s13071-024-06477-1
Huizhu Nan, Xin Lu, Chao Zhang, Xin Yang, Zhu Ying, Lei Ma
Neospora caninum is a protozoan parasite in the Apicomplexa controlled by complex signaling pathways. Transcriptional control, an important way to regulate gene expression, has been almost absent in the N. caninum life process. However, to date, research on the transcriptional regulation of the AP2 family factors in N. caninum has been extremely limited. A prior study demonstrated that removing rhoptry protein 5 (ROP5), a significant virulence factor, resulted in abnormal expression levels of predicted NcAP2XII-4 in N. caninum, suggesting that the factor may regulate the function of ROP5. This study aimed to identify NcAP2XII-4 and its function in transcriptional regulation. The NcAP2XII-4 gene was identified by analyzing the N. caninum genome. A polyclonal antibody against the protein was prepared and purified, and its expression and localization in the parasite were detected using western blot (WB) and immunofluorescence assay (IFA). The ΔNcAP2XII-4 strain was constructed from the Nc1 strain using CRISPR/Cas9 to study its effect on the growth and development of N. caninum, and DAP-Seq and electrophoretic mobility shift assay (EMSA) were used to verify the transcriptional regulatory functions of the gene. Bioinformatic analysis showed that NcAP2XII-4 consists of 11,976 bp and encodes 3991 amino acids, with a predicted molecular mass of 410 kDa. The protein has two AP2 domains, 1207aa-1251aa and 3453aa-3500aa, and is predicted to be located in the nucleus. The results of PCR, WB, and IFA were in accordance with the bioinformatics analysis. ΔNcAP2XII-4 was successfully constructed, but the strain could not be released and ultimately succumbed within parasitophorous vacuoles (PVs). Plaque assays demonstrated that parasites lacking this gene could not form plaques. One motif was successfully identified using DAP-Seq technique. Two prokaryotic expression vectors containing the AP2 domain of NcAP2XII-4 were successfully constructed, and two prokaryotic expression proteins, AP2-D1 and AP2-D2, and ROP5 biotinylated probes were prepared. Using EMSA, NcAP2XII-4 was shown to regulate ROP5 transcription by binding to its promoter. NcAP2XII-4 is an essential gene in N. caninum. This study provides a foundation for further research on transcriptional regulation in N. caninum and identifies a new candidate factor for the development of vaccines against N. caninum.
犬新孢子虫(Neospora caninum)是原生动物寄生虫中的一种,由复杂的信号通路控制。转录调控是调控基因表达的重要途径,但在犬新孢子虫的生命过程中几乎不存在转录调控。然而,迄今为止,对 N. caninum 中 AP2 家族因子转录调控的研究极为有限。之前的一项研究表明,去除 ROPtry 蛋白 5(ROP5)这一重要的毒力因子会导致 N. caninum 中预测的 NcAP2XII-4 的表达水平异常,这表明该因子可能会调控 ROP5 的功能。本研究旨在鉴定 NcAP2XII-4 及其在转录调控中的功能。通过分析 N. caninum 基因组,确定了 NcAP2XII-4 基因。制备并纯化了针对该蛋白的多克隆抗体,并使用免疫印迹(WB)和免疫荧光(IFA)检测了其在寄生虫中的表达和定位。利用CRISPR/Cas9技术在Nc1菌株的基础上构建了ΔNcAP2XII-4菌株,以研究其对犬网虫生长发育的影响,并利用DAP-Seq和电泳迁移分析(EMSA)验证了该基因的转录调控功能。生物信息学分析表明,NcAP2XII-4由11976 bp组成,编码3991个氨基酸,预测分子量为410 kDa。该蛋白有两个 AP2 结构域,分别为 1207aa-1251aa 和 3453aa-3500aa,预计位于细胞核中。PCR、WB和IFA结果与生物信息学分析结果一致。成功构建了ΔNcAP2XII-4,但该菌株无法释放,最终在寄生虫空泡(PVs)中死亡。斑块试验表明,缺乏该基因的寄生虫无法形成斑块。利用 DAP-Seq 技术成功鉴定出了一个基序。成功构建了两种含有 NcAP2XII-4 AP2 结构域的原核表达载体,并制备了 AP2-D1 和 AP2-D2 两种原核表达蛋白以及 ROP5 生物素化探针。通过 EMSA,NcAP2XII-4 与 ROP5 启动子结合,从而调节 ROP5 的转录。NcAP2XII-4 是 N. caninum 的一个重要基因。这项研究为进一步研究犬疫母菌的转录调控奠定了基础,并为开发犬疫母菌疫苗找到了一个新的候选因子。
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引用次数: 0
The spread of the invasive mosquito Aedes albopictus (Diptera: Culicidae) in Portugal: a first genetic analysis 入侵蚊子白纹伊蚊(双翅目:库蚊科)在葡萄牙的传播:首次遗传分析
IF 3.2 2区 医学 Q1 PARASITOLOGY Pub Date : 2024-09-13 DOI: 10.1186/s13071-024-06460-w
Líbia Zé-Zé, Inês Campos Freitas, Manuel Silva, Patrícia Soares, Maria João Alves, Hugo Costa Osório
Aedes albopictus, commonly known as the Asian tiger mosquito, has become one of the most invasive mosquito species. Over the last 5 decades, it has been introduced and established in various tropical and temperate regions worldwide. First reported in Europe in 1979 in Albania and later in Italy in 1990, the species is now established in 13 European Union (EU)/European Economic Area (EEA) countries and 337 regions (2023). In Portugal, Ae. albopictus was first detected in the Algarve and Penafiel regions in 2017, followed by Alentejo in 2022 and Lisbon in 2023. This mosquito species poses a significant public health risk as a vector for numerous pathogenic viruses, including dengue, Zika, and chikungunya. Aedes albopictus collected in Lisbon in 2023 were analyzed using cytochrome c oxidase I (COX) gene sequencing to understand their genetic relationships. Our data indicate that the Ae. albopictus mosquito populations detected in three locations in Lisbon in 2023 correspond to recent but distinct introduction events. Although there has been no local transmission of Aedes-transmitted viruses in mainland Portugal to date, the spread of the mosquito and increased international travel increase the risk of Aedes-borne disease outbreaks. The ongoing spread of Ae. albopictus in the country and the confirmed multiple introductions in new locations raise awareness of the need to monitor mosquito vectors to control and prevent autochthonous Aedes-borne disease outbreaks.
白纹伊蚊,俗称亚洲虎蚊,已成为最具入侵性的蚊子物种之一。在过去的 50 年里,白纹伊蚊已被引入并在世界各地的热带和温带地区定居。该物种于 1979 年首次在欧洲的阿尔巴尼亚被报道,随后于 1990 年在意大利被报道,目前已在 13 个欧盟(EU)/欧洲经济区(EEA)国家和 337 个地区定居(2023 年)。在葡萄牙,白纹伊蚊于 2017 年首次在阿尔加维和佩纳菲尔地区被发现,随后于 2022 年在阿连特茹、2023 年在里斯本被发现。这种蚊子是登革热、寨卡和基孔肯雅等多种致病病毒的传播媒介,对公共卫生构成重大风险。我们使用细胞色素 c 氧化酶 I(COX)基因测序分析了 2023 年在里斯本采集到的白纹伊蚊,以了解它们之间的遗传关系。我们的数据表明,2023 年在里斯本三个地点检测到的白纹伊蚊种群与近期但截然不同的引入事件相对应。虽然迄今为止葡萄牙本土还没有发生伊蚊传播病毒的本地传播,但蚊子的传播和国际旅行的增加增加了伊蚊传播疾病爆发的风险。白纹伊蚊在葡萄牙的持续传播以及在新地点的多次传入都提高了人们对监测蚊媒以控制和预防伊蚊传播疾病爆发的必要性的认识。
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引用次数: 0
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Parasites & Vectors
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