Pub Date : 2019-01-01DOI: 10.21685/1680-0826-2019-13-1-1
A. Tsarev, I. Senderskiy, S. A. Timofeev, V. Zhuravlyov, V. Dolgikh
Obligate parasitism of microsporidia close to fungi protists causing widespread diseases of animals and immunodeficient patients, significantly complicates studies of their relationships with an infected host cell. Since microsporidia cannot be cultivated outside the host cell, genetic manipulations with them are extremely difficult. At the same time, long adaptation of microsporidia to the intracellular lifestyle, drastic minimization of their metabolic machinery, acquisition of unique transporters to exploit a host cell make these parasites a very valuable object for such study and require a search for new methods of investigation. Here, we describe our experiment on the construction of the library of recombinant single chain antibodies (scFv fragments) against proteins of fat bodies of locusts Locusta migratoria infected by the microsporidia Paranosema (Antonospora) locustae. The representativeness of this library was about 108 E. coli transformants carrying different combinations of variable fragments of heavy and light chains of immunized mice immunoglobulins. The first results of the selection of scFv fragments from the constructed library by phage display technology demonstrated that this approach may be effective to search proteins involved in the interaction of microsporidia and other intracellular parasites with an infected host cell. Cloning of selected genes into the expression vector, transformation of E. coli and screening two hundred bacterial colonies revealed scFv fragments against several such candidate proteins to begin their study. Further experiments with the library should discover new variants of recombinant antibodies interacting with the parasite and host proteins.
{"title":"Recombinant single chain antibodies as an instrument to search proteins involved in the interaction of microsporidia and other intracellular parasites with an infected host cell","authors":"A. Tsarev, I. Senderskiy, S. A. Timofeev, V. Zhuravlyov, V. Dolgikh","doi":"10.21685/1680-0826-2019-13-1-1","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-1-1","url":null,"abstract":"Obligate parasitism of microsporidia close to fungi protists causing widespread diseases of animals and immunodeficient patients, significantly complicates studies of their relationships with an infected host cell. Since microsporidia cannot be cultivated outside the host cell, genetic manipulations with them are extremely difficult. At the same time, long adaptation of microsporidia to the intracellular lifestyle, drastic minimization of their metabolic machinery, acquisition of unique transporters to exploit a host cell make these parasites a very valuable object for such study and require a search for new methods of investigation. Here, we describe our experiment on the construction of the library of recombinant single chain antibodies (scFv fragments) against proteins of fat bodies of locusts Locusta migratoria infected by the microsporidia Paranosema (Antonospora) locustae. The representativeness of this library was about 108 E. coli transformants carrying different combinations of variable fragments of heavy and light chains of immunized mice immunoglobulins. The first results of the selection of scFv fragments from the constructed library by phage display technology demonstrated that this approach may be effective to search proteins involved in the interaction of microsporidia and other intracellular parasites with an infected host cell. Cloning of selected genes into the expression vector, transformation of E. coli and screening two hundred bacterial colonies revealed scFv fragments against several such candidate proteins to begin their study. Further experiments with the library should discover new variants of recombinant antibodies interacting with the parasite and host proteins.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67885896","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-2-5
G. Gürelli, Nuray Yürücüoğlu
{"title":"Ruminal ciliate diversity of domestic cattle in Antalya, Turkey, with special emphasis on morphology of Entodinium rostratum (Entodiniomorphida, Ophryoscolecidae)","authors":"G. Gürelli, Nuray Yürücüoğlu","doi":"10.21685/1680-0826-2019-13-2-5","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-2-5","url":null,"abstract":"","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67886600","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-4-4
A. I. Ganyukova, M. N. Malysheva, P. A. Smirnov, A. Frolov
The article provides characteristics of a new parasite, Crithidia dobrovolskii sp.n., which was isolated from the tachinid fly captured in the Leningrad Region of Russia. The presented description of Crithidia dobrovolskii sp.n. is based upon light microscopic, ultrastructural, and molecular phylogenetic data. Molecular phylogenetic analyses of SSU rRNA gene and GAPDH gene sequences have demonstrated that the new species is most closely related to Crithidia fasciculata.
{"title":"Crithidia dobrovolskii sp. n. (Kinetoplastida: Trypanosomatidae) from parasitoid fly Lypha dubia (Diptera: Tachinidae): morphology and phylogenetic position","authors":"A. I. Ganyukova, M. N. Malysheva, P. A. Smirnov, A. Frolov","doi":"10.21685/1680-0826-2019-13-4-4","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-4-4","url":null,"abstract":"The article provides characteristics of a new parasite, Crithidia dobrovolskii sp.n., which was isolated from the tachinid fly captured in the Leningrad Region of Russia. The presented description of Crithidia dobrovolskii sp.n. is based upon light microscopic, ultrastructural, and molecular phylogenetic data. Molecular phylogenetic analyses of SSU rRNA gene and GAPDH gene sequences have demonstrated that the new species is most closely related to Crithidia fasciculata.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67888067","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-2-6
S. Fokin
{"title":"Professor Hans-Dieter Görtz (1945–2019). In memory of my close friend and colleague","authors":"S. Fokin","doi":"10.21685/1680-0826-2019-13-2-6","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-2-6","url":null,"abstract":"","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67886662","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-2-1
O. Matantseva
Dinoflagellates are characterized by the ability to form different types of cysts (resting and temporary) ensuring their viability under unfavorable environmental conditions. Although cyst production was described in many species and is considered as one of the key adaptations of these organisms, relatively little information on the cellular mechanisms of dinoflagellate cyst development and the related process of ecdysis is available. This article reviews the data obtained so far and emphasizes the questions that need to be answered in future in order to improve our knowledge about dinoflagellate physiology and ecological success.
{"title":"Cellular mechanisms of dinoflagellate cyst development and ecdysis – many questions to answer","authors":"O. Matantseva","doi":"10.21685/1680-0826-2019-13-2-1","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-2-1","url":null,"abstract":"Dinoflagellates are characterized by the ability to form different types of cysts (resting and temporary) ensuring their viability under unfavorable environmental conditions. Although cyst production was described in many species and is considered as one of the key adaptations of these organisms, relatively little information on the cellular mechanisms of dinoflagellate cyst development and the related process of ecdysis is available. This article reviews the data obtained so far and emphasizes the questions that need to be answered in future in order to improve our knowledge about dinoflagellate physiology and ecological success.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67886889","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-4-5
Selivanov Ea, M. Ignatenko, T. Yatsenko-Stepanova, A. Plotnikov
Diatoms are highly diverse and widely spread aquatic photosynthetic protists. Studies of regional patterns of diatom diversity are substantial for understanding taxonomy and biogeography of diatoms, as well as for ecological perspectives and applied purposes. DNA barcoding is a modern approach, which can resolve many problems of diatoms identification and can provide valuable information about their diversity in different ecosystems. However, only few studies focused on diatom assemblages of brackish rivers and none of them applied the genetic tools. Herein, we analyzed taxonomic composition and abundance of diatom assemblages in the brackish mixohaline Bolshaya Samoroda River flowing into the Elton Lake (Volgograd region, Russia) using light microscopy and high-throughput sequencing of the V4 region of the 18S rDNA gene amplicons. In total, light microscopy of the samples taken in 2011–2014 and 2018 allowed to distinguish 39 diatom genera, represented by 76 species and infraspecies taxa. Twenty three species of diatoms were recorded in the river for the first time. Next-generation sequencing revealed a larger number of diatom taxa (26 genera and 47 OTUs in two samples vs. 20 genera and 37 species estimated by light microscopy). As a result, sequences of Haslea, Fistulifera, Gedaniella were recorded in the river for the first time. Significant differences in the data obtained with molecular and light microscopy approaches are discussed. Some V4 18S rDNA sequences were characterized by a low similarity with homologues from the reference database. We revealed high spatial-temporal heterogeneity of the diatom assemblages, occurrence of freshwater species together with brackish and marine ones, and predominance of benthic and plankto-benthic species. Thus, investigations of diatoms in brackish rivers based on both morphological and molecular approaches provide a good chance of improving an understanding of diversity, ecology and biogeography of Bacillariophyta.
{"title":"Diatom assemblages of the brackish Bolshaya Samoroda River (Russia) studied via light microscopy and DNA metabarcoding","authors":"Selivanov Ea, M. Ignatenko, T. Yatsenko-Stepanova, A. Plotnikov","doi":"10.21685/1680-0826-2019-13-4-5","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-4-5","url":null,"abstract":"Diatoms are highly diverse and widely spread aquatic photosynthetic protists. Studies of regional patterns of diatom diversity are substantial for understanding taxonomy and biogeography of diatoms, as well as for ecological perspectives and applied purposes. DNA barcoding is a modern approach, which can resolve many problems of diatoms identification and can provide valuable information about their diversity in different ecosystems. However, only few studies focused on diatom assemblages of brackish rivers and none of them applied the genetic tools. Herein, we analyzed taxonomic composition and abundance of diatom assemblages in the brackish mixohaline Bolshaya Samoroda River flowing into the Elton Lake (Volgograd region, Russia) using light microscopy and high-throughput sequencing of the V4 region of the 18S rDNA gene amplicons. In total, light microscopy of the samples taken in 2011–2014 and 2018 allowed to distinguish 39 diatom genera, represented by 76 species and infraspecies taxa. Twenty three species of diatoms were recorded in the river for the first time. Next-generation sequencing revealed a larger number of diatom taxa (26 genera and 47 OTUs in two samples vs. 20 genera and 37 species estimated by light microscopy). As a result, sequences of Haslea, Fistulifera, Gedaniella were recorded in the river for the first time. Significant differences in the data obtained with molecular and light microscopy approaches are discussed. Some V4 18S rDNA sequences were characterized by a low similarity with homologues from the reference database. We revealed high spatial-temporal heterogeneity of the diatom assemblages, occurrence of freshwater species together with brackish and marine ones, and predominance of benthic and plankto-benthic species. Thus, investigations of diatoms in brackish rivers based on both morphological and molecular approaches provide a good chance of improving an understanding of diversity, ecology and biogeography of Bacillariophyta.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"47 7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67888036","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-3-2
Y. Novozhilov, I. Prikhodko, O. Shchepin
A new species of Diderma, described herein as D. dalatense, was found in ground leaf litter in mixed montane tropical forests (Bidoup Nui Ba National Park) of southern Vietnam. This species was discovered during intensive studies on the distribution and ecology of fungi and myxomycetes in tropical forests of Vietnam in the context of a long-term project Ecolan 1.2. The morphology of representative specimens was examined by light and scanning electron microscopy, and micrographs of relevant details are provided. D. dalatense has unique combination of morphological characters of the sporocarps among species of Diderma including very small (0.2–0.4 mm in diam.) dark-yellow, globose to subglobose sessile sporocarps, large conical or subglobose yellow columella, smooth peridium with three layers resembling that of Leocarpus and irregularly warted spores. In addition to the morphological description, partial sequences of three genetic markers of this new species (SSU, EF1A, COI) were obtained and submitted to GenBank. The stability of the taxonomic characters of the species was confirmed by several collections obtained in different localities of the National Park.
{"title":"A new species of Diderma from Bidoup Nui Ba National Park (southern Vietnam)","authors":"Y. Novozhilov, I. Prikhodko, O. Shchepin","doi":"10.21685/1680-0826-2019-13-3-2","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-3-2","url":null,"abstract":"A new species of Diderma, described herein as D. dalatense, was found in ground leaf litter in mixed montane tropical forests (Bidoup Nui Ba National Park) of southern Vietnam. This species was discovered during intensive studies on the distribution and ecology of fungi and myxomycetes in tropical forests of Vietnam in the context of a long-term project Ecolan 1.2. The morphology of representative specimens was examined by light and scanning electron microscopy, and micrographs of relevant details are provided. D. dalatense has unique combination of morphological characters of the sporocarps among species of Diderma including very small (0.2–0.4 mm in diam.) dark-yellow, globose to subglobose sessile sporocarps, large conical or subglobose yellow columella, smooth peridium with three layers resembling that of Leocarpus and irregularly warted spores. In addition to the morphological description, partial sequences of three genetic markers of this new species (SSU, EF1A, COI) were obtained and submitted to GenBank. The stability of the taxonomic characters of the species was confirmed by several collections obtained in different localities of the National Park.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67886887","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-4-1
N. Bondarenko, A. Smirnov, E. Nassonova, A. Glotova, A. Fiore-Donno
In this mini-review, we summarize the current knowledge on mitochondrial genomes of Amoebozoa. Amoebozoa is a major, early-diverging lineage of eukaryotes, containing at least 2,400 species. At present, 32 mitochondrial genomes belonging to 18 amoebozoan species are publicly available. A dearth of information is particularly obvious for two major amoebozoan clades, Variosea and Tubulinea, with just one mitochondrial genome sequenced for each. The main focus of this review is to summarize features such as mitochondrial gene content, mitochondrial genome size variation, and presence or absence of RNA editing, showing if they are unique or shared among amoebozoan lineages. In addition, we underline the potential of mitochondrial genomes for multigene phylogenetic reconstruction in Amoebozoa, where the relationships among lineages are not fully resolved yet. With the increasing application of next-generation sequencing techniques and reliable protocols, we advocate mitochondrial genomes as a promising tool for understanding evolutionary patterns in Amoebozoa.
{"title":"Mitochondrial genomes of Amoebozoa","authors":"N. Bondarenko, A. Smirnov, E. Nassonova, A. Glotova, A. Fiore-Donno","doi":"10.21685/1680-0826-2019-13-4-1","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-4-1","url":null,"abstract":"In this mini-review, we summarize the current knowledge on mitochondrial genomes of Amoebozoa. Amoebozoa is a major, early-diverging lineage of eukaryotes, containing at least 2,400 species. At present, 32 mitochondrial genomes belonging to 18 amoebozoan species are publicly available. A dearth of information is particularly obvious for two major amoebozoan clades, Variosea and Tubulinea, with just one mitochondrial genome sequenced for each. The main focus of this review is to summarize features such as mitochondrial gene content, mitochondrial genome size variation, and presence or absence of RNA editing, showing if they are unique or shared among amoebozoan lineages. In addition, we underline the potential of mitochondrial genomes for multigene phylogenetic reconstruction in Amoebozoa, where the relationships among lineages are not fully resolved yet. With the increasing application of next-generation sequencing techniques and reliable protocols, we advocate mitochondrial genomes as a promising tool for understanding evolutionary patterns in Amoebozoa.","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67887700","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 : 2019-01-01DOI: 10.21685/1680-0826-2019-13-4-6
Olga Gordetskaya, Y. Mesentsev, O. Kamyshatskaya, R. Michel, J. Walochnik, A. Smirnov, E. Nassonova
1 Laboratory of Cytology of Unicellular Organisms, Institute of Cytology RAS, 194064 St. Petersburg, Russia 2 Department of Invertebrate Zoology, Faculty of Biology, St. Petersburg State University, 199034 St. Petersburg, Russia 3 Central Hospital of the Bundeswehr Koblenz, Institute of Pathology, 56065 Koblenz, Germany 4 Institute of Specific Prophylaxis and Tropical Medicine, Medical University of Vienna, 1090 Vienna, Austria
{"title":"Real-time observations on the development of intranuclear parasite Nucleophaga amoebae (Rozellomycota) in the culture of Thecamoeba quadrilineata","authors":"Olga Gordetskaya, Y. Mesentsev, O. Kamyshatskaya, R. Michel, J. Walochnik, A. Smirnov, E. Nassonova","doi":"10.21685/1680-0826-2019-13-4-6","DOIUrl":"https://doi.org/10.21685/1680-0826-2019-13-4-6","url":null,"abstract":"1 Laboratory of Cytology of Unicellular Organisms, Institute of Cytology RAS, 194064 St. Petersburg, Russia 2 Department of Invertebrate Zoology, Faculty of Biology, St. Petersburg State University, 199034 St. Petersburg, Russia 3 Central Hospital of the Bundeswehr Koblenz, Institute of Pathology, 56065 Koblenz, Germany 4 Institute of Specific Prophylaxis and Tropical Medicine, Medical University of Vienna, 1090 Vienna, Austria","PeriodicalId":37502,"journal":{"name":"Protistology","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67888157","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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