Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-2-3
M. N. Malysheva, A. Kostygov, A. Frolov
Tabanids (horseflies and deerflies) represent the main vectors of Trypanosoma theileri species complex and are frequently infected by them. In these insects, the trypanosomes are transiently present in the midgut and develop predominantly in the ileum. During a survey of infections in tabanids, we encountered a horsefly, in which trypanosomatids were present not only in the ileum but also in the rectum. The analysis of 18S rRNA sequences of the parasites in both locations demonstrated that they represented a T. theileri -like trypanosome and the monoxenous species Wallacemonas raviniae previously described from a flesh fly, respectively. The investigation using light and transmission electron microscopy showed that the two parasites differed not only in their affinity to distinct hindgut sections, but also in the patterns of attachment to the cuticular lining, interaction between individual cells, and development of extracellular structures. Unlike most monoxenous trypanosomatids inhabiting the rectum, W. raviniae was not observed on rectal glands or in the close proximity to them indicating either a peculiarity of the parasite species or specific conditions in the insect host. Our results demonstrated that the two trypanosomatid species partitioned their niches within the horsefly host due to strikingly dissimilar life strategies.
{"title":"Niche partitioning within an insect host: trypanosomatids Wallacemonas raviniae and Trypanosoma (Megatrypanum) sp. in the horsefly Hybomitra solstitialis","authors":"M. N. Malysheva, A. Kostygov, A. Frolov","doi":"10.21685/1680-0826-2022-16-2-3","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-2-3","url":null,"abstract":"Tabanids (horseflies and deerflies) represent the main vectors of Trypanosoma theileri species complex and are frequently infected by them. In these insects, the trypanosomes are transiently present in the midgut and develop predominantly in the ileum. During a survey of infections in tabanids, we encountered a horsefly, in which trypanosomatids were present not only in the ileum but also in the rectum. The analysis of 18S rRNA sequences of the parasites in both locations demonstrated that they represented a T. theileri -like trypanosome and the monoxenous species Wallacemonas raviniae previously described from a flesh fly, respectively. The investigation using light and transmission electron microscopy showed that the two parasites differed not only in their affinity to distinct hindgut sections, but also in the patterns of attachment to the cuticular lining, interaction between individual cells, and development of extracellular structures. Unlike most monoxenous trypanosomatids inhabiting the rectum, W. raviniae was not observed on rectal glands or in the close proximity to them indicating either a peculiarity of the parasite species or specific conditions in the insect host. Our results demonstrated that the two trypanosomatid species partitioned their niches within the horsefly host due to strikingly dissimilar life strategies.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67892187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-4-2
M. Tribun
{"title":"Morphology and molecular phylogeny of Euplotes japonicum sp. n. (Ciliophora, Euplotidae) from the Peter the Great Gulf, Sea of Japan","authors":"M. Tribun","doi":"10.21685/1680-0826-2022-16-4-2","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-4-2","url":null,"abstract":"","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67894512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-3-8
A. Kudryavtsev
Summary DNA barcoding using a fragment of the mitochondrial cytochrome c oxidase subunit 1 (Cox1) gene is a promising tool not only in animals but also for many groups of protists, including Amoebozoa. To use this tool, we need a reference database for the comparison and assignment of newly obtained sequences. As NCBI/GeneBank® is the most complete molecular sequence database to date, it is logical to use it as a reference database. In fact, it is used as such, when the newly obtained sequences are checked against this database using BLAST. Yet, a quarter of all available barcoding Cox1 sequences of Amoebozoa would not be seen in the BLAST results, as they are deposited with the status ‘UNVERIFIED’. Some of these sequences show reading frame shifts due to multiple single nucleotide deletions. These deletions, seen at the genomic level, may indicate presence of insertional RNA editing in this gene. This phenomenon was experimentally proven only in myxomycetes and Arcellinida among Amoebozoa. Interestingly, many sequences marked as UNVERIFIED do not show frame shifts or other signs of RNA editing, while some of the sequences that are not assigned this status, do. For the sequence database to be fully searchable, new sequences have to be properly accessioned. A recent communication with NCBI confirms that when a sequence has putative editing sites, the submitter should provide a note on this feature and references to appropriate papers. In this case, the sequence can be accessioned normally.
{"title":"Amoebozoan barcoding marker cytochrome c oxidase (Cox1), RNA editing and issues in creating a public reference sequence database","authors":"A. Kudryavtsev","doi":"10.21685/1680-0826-2022-16-3-8","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-3-8","url":null,"abstract":"Summary DNA barcoding using a fragment of the mitochondrial cytochrome c oxidase subunit 1 (Cox1) gene is a promising tool not only in animals but also for many groups of protists, including Amoebozoa. To use this tool, we need a reference database for the comparison and assignment of newly obtained sequences. As NCBI/GeneBank® is the most complete molecular sequence database to date, it is logical to use it as a reference database. In fact, it is used as such, when the newly obtained sequences are checked against this database using BLAST. Yet, a quarter of all available barcoding Cox1 sequences of Amoebozoa would not be seen in the BLAST results, as they are deposited with the status ‘UNVERIFIED’. Some of these sequences show reading frame shifts due to multiple single nucleotide deletions. These deletions, seen at the genomic level, may indicate presence of insertional RNA editing in this gene. This phenomenon was experimentally proven only in myxomycetes and Arcellinida among Amoebozoa. Interestingly, many sequences marked as UNVERIFIED do not show frame shifts or other signs of RNA editing, while some of the sequences that are not assigned this status, do. For the sequence database to be fully searchable, new sequences have to be properly accessioned. A recent communication with NCBI confirms that when a sequence has putative editing sites, the submitter should provide a note on this feature and references to appropriate papers. In this case, the sequence can be accessioned normally.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"28 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67894556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-4-4
A. Kudryavtsev, E. Volkova, A. Smirnov
a
一个
{"title":"Re-investigation of Vannella ebro Smirnov, 2001 and description of V. navicula sp. nov. (Amoebozoa, Vannellida): further insights into diversity and distribution of marine vannellids","authors":"A. Kudryavtsev, E. Volkova, A. Smirnov","doi":"10.21685/1680-0826-2022-16-4-4","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-4-4","url":null,"abstract":"a","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67894677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-3-6
E. Volkova, F. Voytinsky
{"title":"Description of a new Neoparamoeba aestuarina (Amoebozoa, Dactylopodida) strain from benthic biotopes of the White Sea (Northwestern Russia)","authors":"E. Volkova, F. Voytinsky","doi":"10.21685/1680-0826-2022-16-3-6","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-3-6","url":null,"abstract":"","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67893671","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-3-4
Paul Alain Nana, Nectaire Lié Nyamsi Tchatcho, Fils Mamert Onana, Z. Fokam, C. S. Metsopkeng, P. Ngassam, T. Ewoukem, E. Masseret, M. Nola, T. Sime-Ngando
{"title":"Morphology, morphometry and infraciliature of four new species of Clevelandellida de Puytorac and Grain, 1976 (Ciliophora: Armophorea) from the digestive tube of hydromorphic earthworms in the Cameroonian coastal zone","authors":"Paul Alain Nana, Nectaire Lié Nyamsi Tchatcho, Fils Mamert Onana, Z. Fokam, C. S. Metsopkeng, P. Ngassam, T. Ewoukem, E. Masseret, M. Nola, T. Sime-Ngando","doi":"10.21685/1680-0826-2022-16-3-4","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-3-4","url":null,"abstract":"","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67893576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-4-3
A. Kudryavtsev, E. Volkova, A. Smirnov
Summary Vannella salarenaria sp. nov. was isolated from the upper layer of sand at the dry bottom of Paralimniou lake (Cyprus). This species can be cultured under the salinity up to 40‰, like a typical marine amoeba. However, it also tolerates freshwater conditions and demonstrates a contractile vacuole in this case. Based on the morphological and SSU rRNA data, this species is most closely related to the previously sequenced marine strain Vannella sp. ED40Ana, isolated from the Ebro River delta on the Spanish coast of the Mediterranean Sea in 2000th. We present here archived morphological data on this strain. Among named species, the strain CY18.4SO.2 is most similar morphologically to V. flabellata , described by F.C. Page in 1974 as Platyamoeba . However, it differs from this species in the presence of pentagonal glycostyles in the cell coat. This is the first case of isolation of an amoeba species which can be cultured under marine conditions from the terrestrial habitat. It suggests that amoebae, showing a wide range of salinity tolerance, may populate terrestrial sites in the proximity of the sea.
{"title":"Vannella salarenaria sp. nov. (Amoebozoa, Vannellida) and its implications for the distribution of amoebae","authors":"A. Kudryavtsev, E. Volkova, A. Smirnov","doi":"10.21685/1680-0826-2022-16-4-3","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-4-3","url":null,"abstract":"Summary Vannella salarenaria sp. nov. was isolated from the upper layer of sand at the dry bottom of Paralimniou lake (Cyprus). This species can be cultured under the salinity up to 40‰, like a typical marine amoeba. However, it also tolerates freshwater conditions and demonstrates a contractile vacuole in this case. Based on the morphological and SSU rRNA data, this species is most closely related to the previously sequenced marine strain Vannella sp. ED40Ana, isolated from the Ebro River delta on the Spanish coast of the Mediterranean Sea in 2000th. We present here archived morphological data on this strain. Among named species, the strain CY18.4SO.2 is most similar morphologically to V. flabellata , described by F.C. Page in 1974 as Platyamoeba . However, it differs from this species in the presence of pentagonal glycostyles in the cell coat. This is the first case of isolation of an amoeba species which can be cultured under marine conditions from the terrestrial habitat. It suggests that amoebae, showing a wide range of salinity tolerance, may populate terrestrial sites in the proximity of the sea.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67894612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-1-4
A. I. Ganyukova, M. N. Malysheva, L. Chistyakova, A. Frolov
Summary The trypanosomatid flagellates of the genus Phytomonas were found in the salivary glands and gut of the invasive western conifer seed bug Leptoglossus occidentalis (Hemiptera, Coreidae) in Crimea. The parasites were isolated into axenic culture and identified by phylogenetic analysis based on the 18S rRNA and gGAPDH genes as P. serpens . Previously, all isolates of this species have been obtained in Brazil, from either tomato fruits or their pest bugs Phthia picta (Hemiptera, Coreidae). We investigated the morphology of trypanosomatids from the new isolate in the host bug, laboratory culture, and experimentally infected tomato fruits using light and transmission electron microscopy. We propose hypothetical scenarios of L. occidentalis involvement in the life cycle of P. serpens in the territory, which is new for both species.
{"title":"A puzzling finding: the Brazilian tomato parasite Phytomonas serpens in the western conifer seed bug Leptoglossus occidentalis in Crimea","authors":"A. I. Ganyukova, M. N. Malysheva, L. Chistyakova, A. Frolov","doi":"10.21685/1680-0826-2022-16-1-4","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-1-4","url":null,"abstract":"Summary The trypanosomatid flagellates of the genus Phytomonas were found in the salivary glands and gut of the invasive western conifer seed bug Leptoglossus occidentalis (Hemiptera, Coreidae) in Crimea. The parasites were isolated into axenic culture and identified by phylogenetic analysis based on the 18S rRNA and gGAPDH genes as P. serpens . Previously, all isolates of this species have been obtained in Brazil, from either tomato fruits or their pest bugs Phthia picta (Hemiptera, Coreidae). We investigated the morphology of trypanosomatids from the new isolate in the host bug, laboratory culture, and experimentally infected tomato fruits using light and transmission electron microscopy. We propose hypothetical scenarios of L. occidentalis involvement in the life cycle of P. serpens in the territory, which is new for both species.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"26 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67892295","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2022-01-01DOI: 10.21685/1680-0826-2022-16-1-3
V. Dolgikh, I. Senderskiy, V. Zhuravlyov, Anastasija N. Ignatieva, S. Timofeev, Diloram A. Ismatullaeva, B. A. Mirzakhodjaev
The search for new therapeutics and strategies to suppress microsporidiosis of domesticated insects requires cultivation of honeybee and silkworm parasites in available insect cell cultures as well as reliable methods of their detection. In this study, we infected the commonly used lepidopteran Sf9 cell line with the Vairimorpha ceranae and Nosema bombycis spores to evaluate molecular methods for microsporidia growth assay. The silkworm parasite N. bombycis effectively develops in lepidopteran cells and, according to literature data, its growth can be detected by qPCR analysis of β -tubulin gene copies in infected Sf9 cultures. Here, we used Western blotting with antibodies against N. bombycis β -tubulin to analyze Sf9 cultures infected with the parasite spores and demonstrated the prospects of immunochemical methods to assay its intracellular growth. Analysis of five genes of N. bombycis spore wall and polar tube proteins in infected cultures by reverse transcription (RT) PCR showed that expression of the polar tube protein PTP2 may serve as a specific marker of the parasite growth because only its transcripts were not detected in freshly inoculated Sf9 cells. The honeybee parasite V. ceranae infects Sf9 cells less efficiently. To find a sensitive and specific marker of the growth of this parasite, we analyzed the transcripts of its 13 genes in infected cultures using the same RT-PCR method. The spore wall protein SWP32 gene demonstrated the highest expression at the 4 th -day post infection with V. ceranae spores, alongside with specificity of PCR-amplification, and the absence of transcripts in freshly inoculated cultures. Thus, quantitative PCR analysis of its expression may help to assay the V. ceranae intracellular growth in the Sf9 cell line.
寻找抑制家养昆虫微孢子虫病的新疗法和新策略，需要在现有的昆虫细胞培养物中培养蜜蜂和家蚕寄生虫，以及可靠的检测方法。本研究以鳞翅目常用的Sf9细胞系为研究对象，分别用蜡微孢子虫和家蚕微孢子虫孢子感染鳞翅目Sf9细胞系，探讨微孢子虫生长测定的分子方法。家蚕寄生虫N. bombycis在鳞翅目细胞中有效发育，根据文献资料，通过对感染的Sf9培养物中β -微管蛋白基因拷贝数的qPCR分析，可以检测其生长。本研究采用免疫印迹法(Western blotting)和抗贝氏乳杆菌β -微管蛋白抗体(antibody against N. bombycis β -tubulin)分析了感染贝氏乳杆菌孢子的Sf9培养物，并展示了免疫化学方法检测其细胞内生长的前景。逆转录(RT) PCR分析了感染培养物中家蚕孢子壁和极管蛋白的5个基因，发现在新鲜接种的Sf9细胞中未检测到极管蛋白PTP2的表达，可能是寄生虫生长的特异性标记物。蜜蜂寄生虫V. ceranae感染Sf9细胞的效率较低。为了寻找该寄生虫生长的敏感和特异性标记，我们使用相同的RT-PCR方法分析了感染培养物中其13个基因的转录本。孢子壁蛋白SWP32基因在感染蜡状弧菌孢子第4天时表达量最高，同时具有pcr扩增的特异性，并且在新接种的培养基中没有转录本。因此，对其表达进行定量PCR分析，可能有助于测定丝状弧菌在Sf9细胞株中的胞内生长情况。
{"title":"Molecular detection of microsporidia Vairimorpha ceranae and Nosema bombycis growth in the lepidopteran Sf9 cell line","authors":"V. Dolgikh, I. Senderskiy, V. Zhuravlyov, Anastasija N. Ignatieva, S. Timofeev, Diloram A. Ismatullaeva, B. A. Mirzakhodjaev","doi":"10.21685/1680-0826-2022-16-1-3","DOIUrl":"https://doi.org/10.21685/1680-0826-2022-16-1-3","url":null,"abstract":"The search for new therapeutics and strategies to suppress microsporidiosis of domesticated insects requires cultivation of honeybee and silkworm parasites in available insect cell cultures as well as reliable methods of their detection. In this study, we infected the commonly used lepidopteran Sf9 cell line with the Vairimorpha ceranae and Nosema bombycis spores to evaluate molecular methods for microsporidia growth assay. The silkworm parasite N. bombycis effectively develops in lepidopteran cells and, according to literature data, its growth can be detected by qPCR analysis of β -tubulin gene copies in infected Sf9 cultures. Here, we used Western blotting with antibodies against N. bombycis β -tubulin to analyze Sf9 cultures infected with the parasite spores and demonstrated the prospects of immunochemical methods to assay its intracellular growth. Analysis of five genes of N. bombycis spore wall and polar tube proteins in infected cultures by reverse transcription (RT) PCR showed that expression of the polar tube protein PTP2 may serve as a specific marker of the parasite growth because only its transcripts were not detected in freshly inoculated Sf9 cells. The honeybee parasite V. ceranae infects Sf9 cells less efficiently. To find a sensitive and specific marker of the growth of this parasite, we analyzed the transcripts of its 13 genes in infected cultures using the same RT-PCR method. The spore wall protein SWP32 gene demonstrated the highest expression at the 4 th -day post infection with V. ceranae spores, alongside with specificity of PCR-amplification, and the absence of transcripts in freshly inoculated cultures. Thus, quantitative PCR analysis of its expression may help to assay the V. ceranae intracellular growth in the Sf9 cell line.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67892573","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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