Pub Date : 2024-09-09DOI: 10.1007/s00436-024-08329-4
Martin A. Winkler, Alfred A. Pan
Initial studies using bioinformatics analysis revealed DNA sequence similarities between Trypanosoma cruzi GenBank® M21331, coding for Antigen 36 (Ag 36), and tripartite motif (TRIM) genes. TRIM40 showed 9.7% identity to GenBank M21331, and four additional TRIM genes had identities greater than 5.0%. TRIM37 showed a continuous stretch of identity of 12 nucleotides, that is, at least 25% longer than any of the other TRIMs. When we extended our analysis on the relationships of GenBank M21331 to further innate immune genes, using the Needleman-Wunsch (NW) algorithm for alignment, identities to human IFN-α, IFN-β, and IFN-γ genes of 13.6%, 12.6%, and 17.9%, respectively, were found. To determine the minimum number of genes coding for proteins closely related to Ag 36, a BLAST-p search was conducted with it versus the T. cruzi genome. The BLAST-p search revealed that T. cruzi GenBank M21331 had 14 gene sequences homologous to microtubule-associated protein (MAP) genes with 100% amino acid sequence identity. To verify the similarities in non-human genes, a study comparing TRIM21 region sequences among mammalian species to the comparable human TRIM21 region showed that related sequences were also present in 11 mammalian species. The MAP genes homologous to Ag 36 form a family of at least 14 genes which mimic human immune genes in the IFN and TRIM families. This mimicry is of gene sequences and not their protein products or epitopes. These results appear to be the first description of molecular mimicry of immune genes in humans by a protozoan parasite.
利用生物信息学分析进行的初步研究发现,克氏锥虫 GenBank® M21331(编码抗原 36(Ag 36))和三方基序(TRIM)基因的 DNA 序列具有相似性。TRIM40 与 GenBank M21331 的相同度为 9.7%,另外四个 TRIM 基因的相同度大于 5.0%。TRIM37 显示出 12 个核苷酸的连续同一性,比其他 TRIM 至少长 25%。当我们使用Needleman-Wunsch(NW)算法进行比对时,发现与人类IFN-α、IFN-β和IFN-γ基因的同一性分别为13.6%、12.6%和17.9%。为了确定编码与 Ag 36 密切相关的蛋白质的基因的最低数量,对其与 T. cruzi 基因组进行了 BLAST-p 搜索。BLAST-p 搜索结果显示,T. cruzi GenBank M21331 有 14 个与微管相关蛋白(MAP)基因同源的基因序列,氨基酸序列相同度为 100%。为了验证非人类基因的相似性,一项将哺乳动物物种中的 TRIM21 区域序列与可比较的人类 TRIM21 区域序列进行比较的研究显示,11 种哺乳动物物种中也存在相关序列。与 Ag 36 同源的 MAP 基因组成了一个至少有 14 个基因的家族,这些基因模仿了 IFN 和 TRIM 家族中的人类免疫基因。这种拟态是基因序列的拟态,而不是其蛋白产物或表位的拟态。这些结果似乎是首次描述原生动物寄生虫对人类免疫基因的分子模仿。
{"title":"Molecular similarities between the genes for Trypanosoma cruzi microtubule-associated proteins, mammalian interferons, and TRIMs","authors":"Martin A. Winkler, Alfred A. Pan","doi":"10.1007/s00436-024-08329-4","DOIUrl":"https://doi.org/10.1007/s00436-024-08329-4","url":null,"abstract":"<p>Initial studies using bioinformatics analysis revealed DNA sequence similarities between <i>Trypanosoma cruzi</i> GenBank® M21331, coding for Antigen 36 (Ag 36), and tripartite motif (<i>TRIM</i>) genes. <i>TRIM40</i> showed 9.7% identity to GenBank M21331, and four additional <i>TRIM</i> genes had identities greater than 5.0%. <i>TRIM37</i> showed a continuous stretch of identity of 12 nucleotides, that is, at least 25% longer than any of the other <i>TRIMs</i>. When we extended our analysis on the relationships of GenBank M21331 to further innate immune genes, using the Needleman-Wunsch (NW) algorithm for alignment, identities to human IFN-α, IFN-β, and IFN-γ genes of 13.6%, 12.6%, and 17.9%, respectively, were found. To determine the minimum number of genes coding for proteins closely related to Ag 36, a BLAST-p search was conducted with it versus the <i>T. cruzi</i> genome. The BLAST-p search revealed that <i>T. cruzi</i> GenBank M21331 had 14 gene sequences homologous to microtubule-associated protein (MAP) genes with 100% amino acid sequence identity. To verify the similarities in non-human genes, a study comparing <i>TRIM21</i> region sequences among mammalian species to the comparable human <i>TRIM21</i> region showed that related sequences were also present in 11 mammalian species. The MAP genes homologous to Ag 36 form a family of at least 14 genes which mimic human immune genes in the IFN and <i>TRIM</i> families. This mimicry is of gene sequences and not their protein products or epitopes. These results appear to be the first description of molecular mimicry of immune genes in humans by a protozoan parasite.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"60 1","pages":""},"PeriodicalIF":2.0,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142219800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Several miRNA-based studies on Theileria-transformed bovine cells have been conducted; however, the mechanism by which transformed cells exhibit uncontrolled proliferation is not yet fully understood. Therefore, it is necessary to screen more microRNAs that may play a role in the transformation process of host cells infected with Theileria annulata to better understand the transformation mechanisms of Theileria-infected cells. RNA sequencing was used to analyze miRNAs expression in the host bovine lymphocytes infected with T. annulata at different time points after buparvaquone (BW720) treatment and DMSO treatment (control groups). Differential miRNAs related to cell proliferation and apoptosis were identified through comparison with gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, and a regulatory network of miRNA-mRNA was constructed. In total, 272 differentially expressed miRNAs were found at 36, 60 and 72 h. The miRNAs change of bta-miR-2285t, novel-miR-622, bta-miR-2478, and novel-miR-584 were significant. Analysis of 27 of these co-differential expressed miRNAs revealed that 15 miRNAs were down-regulated and 12 miRNAs were up-regulated. A further analysis of the changes in the expression of each of these 27 miRNAs in the three datasets suggested that bta-miR-2285t, bta-miR-345-5p, bta-miR-34a, bta-miR-150, and the novel-miR-1372 had significantly changed. Predicted target genes for these 27 miRNAs were analyzed by KEGG and the results demonstrated that EZR, RASSF, SOCS1 were mainly enriched in the signaling pathway microRNAs in cancer. MAPKAPK2, RELB, FLT3LG, and GADD45B were mainly enriched in the MAPK signaling pathway, and some genes were enriched in Axon guidance. This study has provided valuable information to further the understanding of the regulatory function of miRNAs in the host microenvironment and host-parasite interaction mechanisms.
{"title":"Expression profile of microRNAs in bovine lymphocytes infected with Theileria annulata and treated with buparvaquone.","authors":"Hong-Xi Zhao, Zhao-Yong Lv, Bao-Cai Zhao, Yue Ma, Xia Li, Gui-Quan Guan","doi":"10.1007/s00436-024-08341-8","DOIUrl":"10.1007/s00436-024-08341-8","url":null,"abstract":"<p><p>Several miRNA-based studies on Theileria-transformed bovine cells have been conducted; however, the mechanism by which transformed cells exhibit uncontrolled proliferation is not yet fully understood. Therefore, it is necessary to screen more microRNAs that may play a role in the transformation process of host cells infected with Theileria annulata to better understand the transformation mechanisms of Theileria-infected cells. RNA sequencing was used to analyze miRNAs expression in the host bovine lymphocytes infected with T. annulata at different time points after buparvaquone (BW720) treatment and DMSO treatment (control groups). Differential miRNAs related to cell proliferation and apoptosis were identified through comparison with gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, and a regulatory network of miRNA-mRNA was constructed. In total, 272 differentially expressed miRNAs were found at 36, 60 and 72 h. The miRNAs change of bta-miR-2285t, novel-miR-622, bta-miR-2478, and novel-miR-584 were significant. Analysis of 27 of these co-differential expressed miRNAs revealed that 15 miRNAs were down-regulated and 12 miRNAs were up-regulated. A further analysis of the changes in the expression of each of these 27 miRNAs in the three datasets suggested that bta-miR-2285t, bta-miR-345-5p, bta-miR-34a, bta-miR-150, and the novel-miR-1372 had significantly changed. Predicted target genes for these 27 miRNAs were analyzed by KEGG and the results demonstrated that EZR, RASSF, SOCS1 were mainly enriched in the signaling pathway microRNAs in cancer. MAPKAPK2, RELB, FLT3LG, and GADD45B were mainly enriched in the MAPK signaling pathway, and some genes were enriched in Axon guidance. This study has provided valuable information to further the understanding of the regulatory function of miRNAs in the host microenvironment and host-parasite interaction mechanisms.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"318"},"PeriodicalIF":1.8,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142154824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-05DOI: 10.1007/s00436-024-08333-8
Augustus M Snyder, Eric J McElroy, Isaure de Buron, Fabio Casu, Jody M Beers
The spotted seatrout, Cynoscion nebulosus, is a popular game fish in the southeastern USA. It is estimated that nearly 90% of the adult population in South Carolina estuaries are infected in their skeletal muscle by the myxosporean, Kudoa inornata. However, little is known about this parasite's biology, including the distribution and densities of myxospores within tissues of infected fish, which we expect affect the physiology of their hosts. In order to correlate densities with physiological parameters in future studies, we quantified the myxospores density in muscle and characterized the variation among individual fish. Naïve juvenile seatrout was experimentally infected via presumed K. inornata actinospores exposure to raw seawater. A plug of muscle was extracted from two bilaterally symmetrical regions in the epaxial fillet from fresh and frozen carcasses. Variation in density data was calculated both within and among individuals. Within individuals, density counts were compared between left- and right-side biopsies. There was no significant difference between fresh and frozen plugs, and variation among individuals accounted for the greatest proportion of variation at 68.8%, while variation within individuals was substantial at 25.6%. Simulation and correlation tests confirmed that bilaterally symmetrical replicates varied significantly within individuals. When sampled from areas surrounding the initial biopsies, myxospore density estimates were more similar than between sides. Our findings have important implications for sampling design, particularly for studies investigating physiological parameters at the cellular or molecular level in association with parasite infection.
{"title":"Myxospore density of Kudoa inornata varies significantly within symmetrical white muscle tissue replicates of its fish host, the spotted seatrout, Cynoscion nebulosus.","authors":"Augustus M Snyder, Eric J McElroy, Isaure de Buron, Fabio Casu, Jody M Beers","doi":"10.1007/s00436-024-08333-8","DOIUrl":"10.1007/s00436-024-08333-8","url":null,"abstract":"<p><p>The spotted seatrout, Cynoscion nebulosus, is a popular game fish in the southeastern USA. It is estimated that nearly 90% of the adult population in South Carolina estuaries are infected in their skeletal muscle by the myxosporean, Kudoa inornata. However, little is known about this parasite's biology, including the distribution and densities of myxospores within tissues of infected fish, which we expect affect the physiology of their hosts. In order to correlate densities with physiological parameters in future studies, we quantified the myxospores density in muscle and characterized the variation among individual fish. Naïve juvenile seatrout was experimentally infected via presumed K. inornata actinospores exposure to raw seawater. A plug of muscle was extracted from two bilaterally symmetrical regions in the epaxial fillet from fresh and frozen carcasses. Variation in density data was calculated both within and among individuals. Within individuals, density counts were compared between left- and right-side biopsies. There was no significant difference between fresh and frozen plugs, and variation among individuals accounted for the greatest proportion of variation at 68.8%, while variation within individuals was substantial at 25.6%. Simulation and correlation tests confirmed that bilaterally symmetrical replicates varied significantly within individuals. When sampled from areas surrounding the initial biopsies, myxospore density estimates were more similar than between sides. Our findings have important implications for sampling design, particularly for studies investigating physiological parameters at the cellular or molecular level in association with parasite infection.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"317"},"PeriodicalIF":1.8,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11377670/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142133447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Schistosomiasis remains a formidable challenge to global public health. This study aims to predict the spatial distribution of schistosomiasis seropositive rates in Hunan Province, pinpointing high-risk transmission areas and advocating for tailored control measures in low-endemic regions. Six machine learning models and their corresponding hybrid machine learning-Kriging models were employed to predict the seropositive rate. The optimal model was selected through internal and external validations to simulate the spatial distribution of seropositive rates. Our results showed that the hybrid machine learning-Kriging model demonstrated superior predictive performance compared to basic machine learning model and the Cubist-Kriging model emerged as the most optimal model for this study. The predictive map revealed elevated seropositive rates around Dongting Lake and its waterways with significant clustering, notably in the central and northern regions of Yiyang City and the northeastern areas of Changde City. The model identified gross domestic product, annual average wind speed and the nearest distance from the river as the top three predictors of seropositive rates, with annual average daytime surface temperature contributing the least. In conclusion, our research has revealed that integrating the Kriging method significantly enhances the predictive performance of machine learning models. We developed a Cubist-Kriging model with high predictive performance to forecast the spatial distribution of schistosomiasis seropositive rates. These findings provide valuable guidance for the precise prevention and control of schistosomiasis.
{"title":"Prediction on the spatial distribution of the seropositive rate of schistosomiasis in Hunan Province, China: a machine learning model integrated with the Kriging method.","authors":"Ning Xu, Yu Cai, Yixin Tong, Ling Tang, Yu Zhou, Yanfeng Gong, Junhui Huang, Jiamin Wang, Yue Chen, Qingwu Jiang, Mao Zheng, Yibiao Zhou","doi":"10.1007/s00436-024-08331-w","DOIUrl":"10.1007/s00436-024-08331-w","url":null,"abstract":"<p><p>Schistosomiasis remains a formidable challenge to global public health. This study aims to predict the spatial distribution of schistosomiasis seropositive rates in Hunan Province, pinpointing high-risk transmission areas and advocating for tailored control measures in low-endemic regions. Six machine learning models and their corresponding hybrid machine learning-Kriging models were employed to predict the seropositive rate. The optimal model was selected through internal and external validations to simulate the spatial distribution of seropositive rates. Our results showed that the hybrid machine learning-Kriging model demonstrated superior predictive performance compared to basic machine learning model and the Cubist-Kriging model emerged as the most optimal model for this study. The predictive map revealed elevated seropositive rates around Dongting Lake and its waterways with significant clustering, notably in the central and northern regions of Yiyang City and the northeastern areas of Changde City. The model identified gross domestic product, annual average wind speed and the nearest distance from the river as the top three predictors of seropositive rates, with annual average daytime surface temperature contributing the least. In conclusion, our research has revealed that integrating the Kriging method significantly enhances the predictive performance of machine learning models. We developed a Cubist-Kriging model with high predictive performance to forecast the spatial distribution of schistosomiasis seropositive rates. These findings provide valuable guidance for the precise prevention and control of schistosomiasis.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"316"},"PeriodicalIF":1.8,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142126381","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-04DOI: 10.1007/s00436-024-08338-3
Yu Tang, Heng Li, Qiaoling Song
Mosquito-borne diseases, such as malaria, dengue fever, and the Zika virus, pose significant global health challenges, affecting millions annually. Due to increasing insecticide resistance, there is a growing interest in natural alternatives for mosquito control. Lemongrass essential oil, derived from Cymbopogon citratus, has shown promising repellent and larvicidal properties against various mosquito species. In this study, we investigated the larvicidal effect of lemongrass oil and its major compounds on Anopheles sinensis, the primary malaria vector in China. GC-MS analysis identified the major compounds of lemongrass oil as ( +)-citronellal (35.60%), geraniol (21.84%), and citronellol (13.88%). Lemongrass oil showed larvicidal activity against An. sinensis larvae, with an LC50 value of 119.20 ± 3.81 mg/L. Among the major components, citronellol had the lowest LC50 value of 42.76 ± 3.18 mg/L. Moreover, citronellol demonstrated inhibitory effects on acetylcholinesterase (AChE) activity in An. sinensis larvae, assessed by homogenizing larvae at different time points following treatment. Molecular docking studies further elucidated the interaction between citronellol and AChE, revealing the formation of hydrogen bonds and Pi-Sigma bonds. Aromatic amino acid residues such as Tyr71, Trp83, Tyr370, and Tyr374 played a pivotal role in these interactions. These findings may contribute to understanding lemongrass oil's larvicidal activity against An. sinensis and the mechanisms underlying these effects.
{"title":"Lemongrass essential oil and its major component citronellol: evaluation of larvicidal activity and acetylcholinesterase inhibition against Anopheles sinensis.","authors":"Yu Tang, Heng Li, Qiaoling Song","doi":"10.1007/s00436-024-08338-3","DOIUrl":"10.1007/s00436-024-08338-3","url":null,"abstract":"<p><p>Mosquito-borne diseases, such as malaria, dengue fever, and the Zika virus, pose significant global health challenges, affecting millions annually. Due to increasing insecticide resistance, there is a growing interest in natural alternatives for mosquito control. Lemongrass essential oil, derived from Cymbopogon citratus, has shown promising repellent and larvicidal properties against various mosquito species. In this study, we investigated the larvicidal effect of lemongrass oil and its major compounds on Anopheles sinensis, the primary malaria vector in China. GC-MS analysis identified the major compounds of lemongrass oil as ( +)-citronellal (35.60%), geraniol (21.84%), and citronellol (13.88%). Lemongrass oil showed larvicidal activity against An. sinensis larvae, with an LC<sub>50</sub> value of 119.20 ± 3.81 mg/L. Among the major components, citronellol had the lowest LC<sub>50</sub> value of 42.76 ± 3.18 mg/L. Moreover, citronellol demonstrated inhibitory effects on acetylcholinesterase (AChE) activity in An. sinensis larvae, assessed by homogenizing larvae at different time points following treatment. Molecular docking studies further elucidated the interaction between citronellol and AChE, revealing the formation of hydrogen bonds and Pi-Sigma bonds. Aromatic amino acid residues such as Tyr71, Trp83, Tyr370, and Tyr374 played a pivotal role in these interactions. These findings may contribute to understanding lemongrass oil's larvicidal activity against An. sinensis and the mechanisms underlying these effects.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"315"},"PeriodicalIF":1.8,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142126380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-03DOI: 10.1007/s00436-024-08330-x
Amanda Martins Ungri, Bruna Fernanda Dos Santos Sabatke, Izadora Volpato Rossi, Gabriella Bassi das Neves, Júlia Marques, Brenda Guedes Ribeiro, Gabriela Kaiser Borges, Renato Simões Moreira, Marcel Ivan Ramírez, Luiz Claudio Miletti
Trypanosoma evansi is a unicellular protozoan responsible for causing a disease known as "surra," which is found in different regions of the world and primarily affects horses and camels. Few information is known about virulence factors released from the parasite within the animals. The organism can secrete extracellular vesicles (EVs), which transport a variety of molecules, including proteins. Before being considered exclusively as a means for eliminating unwanted substances, extracellular vesicles (EVs) have emerged as key players in intercellular communication, facilitating interactions between cells, host cells, and parasites, and even between parasites themselves. Thus, they may be used as potential biomarkers. This study aimed to assess the induction of EVs production by Ca+2, conduct a proteomic analysis of the EVs released by T. evansi, and identify epitopes that could serve as biomarkers. The findings indicated that Ca+2 is not an effective promoter of vesiculation in T. evansi. Furthermore, the proteomic analysis has identified multiple proteins that have been investigated as biomarkers or vaccine antigens, previously. A total of 442 proteins were identified, with 7 of them specifically recognizing 9 epitopes that are unique to T. evansi. At least one of these epitopes of TevSTIB805.9.11580 have been previously identified, which increases the possibility of further investigating its potential as a biomarker.
{"title":"Extracellular vesicles released by Trypanosoma evansi: induction analysis and proteomics.","authors":"Amanda Martins Ungri, Bruna Fernanda Dos Santos Sabatke, Izadora Volpato Rossi, Gabriella Bassi das Neves, Júlia Marques, Brenda Guedes Ribeiro, Gabriela Kaiser Borges, Renato Simões Moreira, Marcel Ivan Ramírez, Luiz Claudio Miletti","doi":"10.1007/s00436-024-08330-x","DOIUrl":"10.1007/s00436-024-08330-x","url":null,"abstract":"<p><p>Trypanosoma evansi is a unicellular protozoan responsible for causing a disease known as \"surra,\" which is found in different regions of the world and primarily affects horses and camels. Few information is known about virulence factors released from the parasite within the animals. The organism can secrete extracellular vesicles (EVs), which transport a variety of molecules, including proteins. Before being considered exclusively as a means for eliminating unwanted substances, extracellular vesicles (EVs) have emerged as key players in intercellular communication, facilitating interactions between cells, host cells, and parasites, and even between parasites themselves. Thus, they may be used as potential biomarkers. This study aimed to assess the induction of EVs production by Ca<sup>+2</sup>, conduct a proteomic analysis of the EVs released by T. evansi, and identify epitopes that could serve as biomarkers. The findings indicated that Ca<sup>+2</sup> is not an effective promoter of vesiculation in T. evansi. Furthermore, the proteomic analysis has identified multiple proteins that have been investigated as biomarkers or vaccine antigens, previously. A total of 442 proteins were identified, with 7 of them specifically recognizing 9 epitopes that are unique to T. evansi. At least one of these epitopes of TevSTIB805.9.11580 have been previously identified, which increases the possibility of further investigating its potential as a biomarker.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"314"},"PeriodicalIF":1.8,"publicationDate":"2024-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142120431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-02DOI: 10.1007/s00436-024-08327-6
Joban Quesada, Paula Alfaro-Segura, Carlos Mata-Somarribas, Jackeline Alger, Mazlova Toledo, Jucicleide Ramos de Souza, Javier Mora, Carlos Graeff-Teixeira, Alberto Solano-Barquero, Alicia Rojas
Abdominal angiostrongyliasis (AA) is a zoonotic and severe parasitic infection caused by Angiostrongylus costaricensis. AA is currently diagnosed by the observation of A. costaricensis-compatible structures in biopsies or the detection of antibodies in serological tests. However, molecular methods targeting homologous sequences of A. costaricensis have not been designed before, and therefore, an HRM-coupled qPCR was developed to detect the internal transcribed spacer 1 (ITS1) of the parasite. The present assay successfully amplified DNA of A. costaricensis obtained from different hosts and identified slight sequence differences through the HRM analysis. The detection limit of the HRM-qPCR was 0.00036 ng/µL, 1.0 ng/µL, and 0.1 ng/µL when A. costaricensis DNA was diluted in nuclease-free water, whole blood, and sera, respectively, which highlights its potential use for cell-free DNA detection. Moreover, the reaction did not cross-amplify DNA of Angiostrongylus cantonensis, Strongyloides stercoralis, and other nematodes, thus emphasizing its specificity. Additionally, the assay tested positive in formalin-fixed paraffin embedded biopsies with visible A. costaricensis adults or eggs, but not in samples without evident parasites or a low number of larvae, which suggests that the reaction is useful for confirming the presence of the nematode in clinical samples. Finally, DNA of sera from patients with AA was evaluated with the HRM-qPCR but none tested positive, possibly due to long storage periods of the samples which could have led to cfDNA degradation. These results indicate that this assay may be useful in the confirmation of AA and its prospection for cell-free DNA detection protocols.
腹腔角弓反张症(AA)是由腹腔角弓反张蚤(Angiostrongylus costaricensis)引起的人畜共患的严重寄生虫病。目前,AA 的诊断方法是在活组织切片中观察到与 A. costaricensis 相符的结构,或在血清检验中检测到抗体。因此,我们开发了一种 HRM 偶联 qPCR 方法来检测寄生虫的内部转录间隔 1(ITS1)。本检测方法成功地扩增了从不同宿主处获得的 A. costaricensis DNA,并通过 HRM 分析确定了轻微的序列差异。当 A. costaricensis DNA 被稀释在无核酸水、全血和血清中时,HRM-qPCR 的检测限分别为 0.00036 ng/µL、1.0 ng/µL 和 0.1 ng/µL,这突显了其用于无细胞 DNA 检测的潜力。此外,该反应不会交叉扩增坎顿角弓形虫、盘尾丝虫和其他线虫的 DNA,因此强调了其特异性。此外,在福尔马林固定的石蜡包埋活检样本中,该检测方法对可见的库氏安氏线虫成虫或虫卵检测呈阳性,而对无明显寄生虫或幼虫数量较少的样本检测呈阳性,这表明该反应可用于确认临床样本中是否存在线虫。最后,用 HRM-qPCR 对 AA 患者血清中的 DNA 进行了评估,但检测结果均为阳性,这可能是由于样本储存时间过长导致 cfDNA 降解所致。这些结果表明,该检测方法可用于 AA 的确诊,并有望用于无细胞 DNA 检测方案。
{"title":"Real-time qPCR coupled with high-resolution melting curve analysis for the detection of the internal transcribed spacer 1 of Angiostrongylus costaricensis.","authors":"Joban Quesada, Paula Alfaro-Segura, Carlos Mata-Somarribas, Jackeline Alger, Mazlova Toledo, Jucicleide Ramos de Souza, Javier Mora, Carlos Graeff-Teixeira, Alberto Solano-Barquero, Alicia Rojas","doi":"10.1007/s00436-024-08327-6","DOIUrl":"10.1007/s00436-024-08327-6","url":null,"abstract":"<p><p>Abdominal angiostrongyliasis (AA) is a zoonotic and severe parasitic infection caused by Angiostrongylus costaricensis. AA is currently diagnosed by the observation of A. costaricensis-compatible structures in biopsies or the detection of antibodies in serological tests. However, molecular methods targeting homologous sequences of A. costaricensis have not been designed before, and therefore, an HRM-coupled qPCR was developed to detect the internal transcribed spacer 1 (ITS1) of the parasite. The present assay successfully amplified DNA of A. costaricensis obtained from different hosts and identified slight sequence differences through the HRM analysis. The detection limit of the HRM-qPCR was 0.00036 ng/µL, 1.0 ng/µL, and 0.1 ng/µL when A. costaricensis DNA was diluted in nuclease-free water, whole blood, and sera, respectively, which highlights its potential use for cell-free DNA detection. Moreover, the reaction did not cross-amplify DNA of Angiostrongylus cantonensis, Strongyloides stercoralis, and other nematodes, thus emphasizing its specificity. Additionally, the assay tested positive in formalin-fixed paraffin embedded biopsies with visible A. costaricensis adults or eggs, but not in samples without evident parasites or a low number of larvae, which suggests that the reaction is useful for confirming the presence of the nematode in clinical samples. Finally, DNA of sera from patients with AA was evaluated with the HRM-qPCR but none tested positive, possibly due to long storage periods of the samples which could have led to cfDNA degradation. These results indicate that this assay may be useful in the confirmation of AA and its prospection for cell-free DNA detection protocols.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"312"},"PeriodicalIF":1.8,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142110800","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The practice of hybridization is carried out globally in fish farms. Here, we present the first record of the parasitic fauna of hybrids among genus Colossoma and Piaractus in natural environments. We identified a total of 48 hybrids, nine F1 hybrids (nuclear DNA from both species present in the cross) and 38 advanced hybrids (nuclear DNA from one species), both from crosses between Piaractus brachypomus and Piaractus mesopotamicus, and one F1 "tambacu" corresponding to cross between Colossoma macropomum and Piaractus mesopotamicus. This is the first record of Anacanthorus penilabiatus, Anacanthorus toledoensis, Mymarothecium viatorum, Mymarothecium ianwhittington, Haementeria sp., Dadaytrema oxycephala, Rondonia rondoni, and Echinorhynchus gomesi parasitizing hybrids collected in a natural environment. With this, we expand knowledge about the diversity of fish and parasites in the upper Paraná River and warn about the risk that fish escapes can cause in the basin.
{"title":"First record of metazoan parasites of hybrids between the genera Colossoma and Piaractus in natural environments.","authors":"Lidiany Doreto Cavalcanti, Gabriela Michelan, Wagner Toshio Hasuike, João Otávio Santos Silva, Atsler Luana Lehun, Eloisa Balabuch, Bruna Fernanda Rodrigues Bandeira, Márcia Regina Russo, Ricardo Massato Takemoto","doi":"10.1007/s00436-024-08335-6","DOIUrl":"10.1007/s00436-024-08335-6","url":null,"abstract":"<p><p>The practice of hybridization is carried out globally in fish farms. Here, we present the first record of the parasitic fauna of hybrids among genus Colossoma and Piaractus in natural environments. We identified a total of 48 hybrids, nine F1 hybrids (nuclear DNA from both species present in the cross) and 38 advanced hybrids (nuclear DNA from one species), both from crosses between Piaractus brachypomus and Piaractus mesopotamicus, and one F1 \"tambacu\" corresponding to cross between Colossoma macropomum and Piaractus mesopotamicus. This is the first record of Anacanthorus penilabiatus, Anacanthorus toledoensis, Mymarothecium viatorum, Mymarothecium ianwhittington, Haementeria sp., Dadaytrema oxycephala, Rondonia rondoni, and Echinorhynchus gomesi parasitizing hybrids collected in a natural environment. With this, we expand knowledge about the diversity of fish and parasites in the upper Paraná River and warn about the risk that fish escapes can cause in the basin.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"313"},"PeriodicalIF":1.8,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142110798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-30DOI: 10.1007/s00436-024-08334-7
Hannes Wickert, Georg Krohne
Striking morphological transformations characterize the invasion of a red blood cell by the malaria parasite. Shortly after the infection, parasite-induced membranes appear in the cytosol of the affected host erythrocyte. One intensely investigated membrane type, commonly called Maurer's clefts, has a slit-like morphology and can be arranged in the form of extended three-dimensional membrane stacks or networks. Here we report the three-dimensional reconstruction of a second membrane type, giant or extended membrane rings/loops, that have only occasionally been described on single ultrathin sections, however that have never been systematically examined so far. Serial ultrathin sectioning of P. falciparum-infected red blood cells, subsequent three-dimensional reconstructions, and in addition examination of Giemsa-stained blood films revealed that intraerythrocytic membrane rings/loops are not isolated structures but are locally in contact with the parasite. They consist either of the parasitophorous vacuolar membrane alone or contain the parasitophorous vacuolar membrane including the plasma membrane of the parasite and small amounts of parasite cytoplasm. We demonstrate that membrane rings/loops represent surface extensions of the parasite that maybe involved in ring stage parasite formation and Maurer's cleft generation at least in a subset of infected red blood cells.
{"title":"Giant membrane rings/loops in the cytosol of P. falciparum-infected erythrocytes and their relation to the parasite.","authors":"Hannes Wickert, Georg Krohne","doi":"10.1007/s00436-024-08334-7","DOIUrl":"10.1007/s00436-024-08334-7","url":null,"abstract":"<p><p>Striking morphological transformations characterize the invasion of a red blood cell by the malaria parasite. Shortly after the infection, parasite-induced membranes appear in the cytosol of the affected host erythrocyte. One intensely investigated membrane type, commonly called Maurer's clefts, has a slit-like morphology and can be arranged in the form of extended three-dimensional membrane stacks or networks. Here we report the three-dimensional reconstruction of a second membrane type, giant or extended membrane rings/loops, that have only occasionally been described on single ultrathin sections, however that have never been systematically examined so far. Serial ultrathin sectioning of P. falciparum-infected red blood cells, subsequent three-dimensional reconstructions, and in addition examination of Giemsa-stained blood films revealed that intraerythrocytic membrane rings/loops are not isolated structures but are locally in contact with the parasite. They consist either of the parasitophorous vacuolar membrane alone or contain the parasitophorous vacuolar membrane including the plasma membrane of the parasite and small amounts of parasite cytoplasm. We demonstrate that membrane rings/loops represent surface extensions of the parasite that maybe involved in ring stage parasite formation and Maurer's cleft generation at least in a subset of infected red blood cells.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 9","pages":"311"},"PeriodicalIF":1.8,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142110799","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-29DOI: 10.1007/s00436-024-08328-5
Fangjie Li, Jiaying Guo, Sen Wang, Zhen Han, Zheng Nie, Long Yu, Xiang Shu, Yingjun Xia, Lan He, Junlong Zhao
Babesia orientalis, a protozoan parasite transmitted by the tick Rhipicephalus haemaphysaloides, holds significant economic importance along the Yangtze River. Key factors in the host invasion process include rhoptry neck proteins (RON2, RON4, and RON5) and apical membrane antigen 1 (AMA1). However, the intricacies of the interaction between AMA1 and RONs remain incompletely elucidated in B. orientalis. To better understand these crucial invasion components, the RON4 gene of B. orientalis (BoRON4) was cloned and sequenced. RON4 is 3468 base pairs long, encodes 1155 amino acids, and has a predicted molecular weight of 130 kDa. Bioinformatics analysis revealed a unique region (amino acid residues 109-452) in BoRON4, which demonstrates higher sensitivity to epitope activity. The BoRON4 gene was strategically truncated, amplified, and cloned into the pGEX-6p-1 vector for fusion expression. We successfully used the mouse polyclonal antibody to identify native BoRON4 in B. orientalis lysates. Furthermore, the corresponding BoRON4 protein band was detected in the water buffalo serum infected with B. orientalis, while no such band was observed in the control. Additionally, I-TASSER and Discovery Studio software were used to predict the tertiary structures of BoRON4 and its ligands, CH-PKA and CH-complex. These ligands can serve as lead compounds for the development of anti-babesiosis drugs. In conclusion, BoRON4 emerges as a promising candidate antigen for distinguishing water buffalo infected with B. orientalis from their normal counterparts. This study positions BoRON4 as a potential diagnostic antigen for babesiosis in water buffalo, contributing valuable insights to the field of parasitology.
{"title":"Identification and molecular characterization of a novel Babesia orientalis rhoptry neck protein 4 (BoRON4).","authors":"Fangjie Li, Jiaying Guo, Sen Wang, Zhen Han, Zheng Nie, Long Yu, Xiang Shu, Yingjun Xia, Lan He, Junlong Zhao","doi":"10.1007/s00436-024-08328-5","DOIUrl":"10.1007/s00436-024-08328-5","url":null,"abstract":"<p><p>Babesia orientalis, a protozoan parasite transmitted by the tick Rhipicephalus haemaphysaloides, holds significant economic importance along the Yangtze River. Key factors in the host invasion process include rhoptry neck proteins (RON2, RON4, and RON5) and apical membrane antigen 1 (AMA1). However, the intricacies of the interaction between AMA1 and RONs remain incompletely elucidated in B. orientalis. To better understand these crucial invasion components, the RON4 gene of B. orientalis (BoRON4) was cloned and sequenced. RON4 is 3468 base pairs long, encodes 1155 amino acids, and has a predicted molecular weight of 130 kDa. Bioinformatics analysis revealed a unique region (amino acid residues 109-452) in BoRON4, which demonstrates higher sensitivity to epitope activity. The BoRON4 gene was strategically truncated, amplified, and cloned into the pGEX-6p-1 vector for fusion expression. We successfully used the mouse polyclonal antibody to identify native BoRON4 in B. orientalis lysates. Furthermore, the corresponding BoRON4 protein band was detected in the water buffalo serum infected with B. orientalis, while no such band was observed in the control. Additionally, I-TASSER and Discovery Studio software were used to predict the tertiary structures of BoRON4 and its ligands, CH-PKA and CH-complex. These ligands can serve as lead compounds for the development of anti-babesiosis drugs. In conclusion, BoRON4 emerges as a promising candidate antigen for distinguishing water buffalo infected with B. orientalis from their normal counterparts. This study positions BoRON4 as a potential diagnostic antigen for babesiosis in water buffalo, contributing valuable insights to the field of parasitology.</p>","PeriodicalId":19968,"journal":{"name":"Parasitology Research","volume":"123 8","pages":"310"},"PeriodicalIF":1.8,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142110797","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}