Trypanosomatids of the subfamily Strigomonadinae bear permanent intracellular bacterial symbionts acquired by the common ancestor of these flagellates. However, the cospeciation pattern inherent to such relationships was revealed to be broken upon the description of Angomonas ambiguus, which is sister to A. desouzai, but bears an endosymbiont genetically close to that of A. deanei. Based on phylogenetic inferences, it was proposed that the bacterium from A. deanei had been horizontally transferred to A. ambiguus. Here, we sequenced the bacterial genomes from two A. ambiguus isolates, including a new one from Papua New Guinea, and compared them with the published genome of the A. deanei endosymbiont, revealing differences below the interspecific level. Our phylogenetic analyses confirmed that the endosymbionts of A. ambiguus were obtained from A. deanei and, in addition, demonstrated that this occurred more than once. We propose that coinfection of the same blowfly host and the phylogenetic relatedness of the trypanosomatids facilitate such transitions, whereas the drastic difference in the occurrence of the two trypanosomatid species determines the observed direction of this process. This phenomenon is analogous to organelle (mitochondrion/plastid) capture described in multicellular organisms and, thereafter, we name it endosymbiont capture.

• Klíčová slova
• Angomonas, Trypanosomatidae, bacterial endosymbionts, genome,
• Publikační typ
• časopisecké články MeSH

Parasitic trypanosomatids diverged from free-living kinetoplastid ancestors several hundred million years ago. These parasites are relatively well known, due in part to several unusual cell biological and molecular traits and in part to the significance of a few - pathogenic Leishmania and Trypanosoma species - as aetiological agents of serious neglected tropical diseases. However, the majority of trypanosomatid biodiversity is represented by osmotrophic monoxenous parasites of insects. In two lineages, novymonads and strigomonads, osmotrophic lifestyles are supported by cytoplasmic endosymbionts, providing hosts with macromolecular precursors and vitamins. Here we discuss the two independent origins of endosymbiosis within trypanosomatids and subsequently different evolutionary trajectories that see entrainment vs tolerance of symbiont cell divisions cycles within those of the host. With the potential to inform on the transition to obligate parasitism in the trypanosomatids, interest in the biology and ecology of free-living, phagotrophic kinetoplastids is beginning to enjoy a renaissance. Thus, we take the opportunity to additionally consider the wider relevance of endosymbiosis during kinetoplastid evolution, including the indulged lifestyle and reductive evolution of basal kinetoplastid Perkinsela.

• Klíčová slova
• Angomonas deanei, Candidatus Kinetoplastibacterium, Kentomonas, Novymonas esmeraldas, Pandoraea, cytostome,
• MeSH
• biodiverzita MeSH
• biologická evoluce * MeSH
• genom protozoální MeSH
• Kinetoplastida genetika   MeSH
• Leishmania genetika   fyziologie   MeSH
• molekulární evoluce MeSH
• symbióza * MeSH
• Trypanosoma genetika   fyziologie   MeSH
• Trypanosomatina genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Trypanosomatids are obligate parasites of animals, predominantly insects and vertebrates, and flowering plants. Monoxenous species, representing the vast majority of trypanosomatid diversity, develop in a single host, whereas dixenous species cycle between two hosts, of which primarily insect serves as a vector. To explore in-depth the diversity of insect trypanosomatids including their co-infections, sequence profiling of their 18S rRNA gene was used for true bugs (Hemiptera; 18% infection rate) and flies (Diptera; 10%) in Cuba. Out of 48 species (molecular operational taxonomic units) belonging to the genera Vickermania (16 spp.), Blastocrithidia (7), Obscuromonas (4), Phytomonas (5), Leptomonas/Crithidia (5), Herpetomonas (5), Wallacemonas (2), Kentomonas (1), Angomonas (1) and two unnamed genera (1 + 1), 38 species have been encountered for the first time. The detected Wallacemonas and Angomonas species constitute the most basal lineages of their respective genera, while Vickermania emerged as the most diverse group. The finding of Leptomonas seymouri, which is known to rarely infect humans, confirms that Dysdercus bugs are its natural hosts. A clear association of Phytomonas with the heteropteran family Pentatomidae hints at its narrow host association with the insect rather than plant hosts. With a focus on multiple infections of a single fly host, using deep Nanopore sequencing of 18S rRNA, we have identified co-infections with up to 8 trypanosomatid species. The fly midgut was usually occupied by several Vickermania species, while Herpetomonas and/or Kentomonas species prevailed in the hindgut. Metabarcoding was instrumental for analysing extensive co-infections and also allowed the identification of trypanosomatid lineages and genera.

• Klíčová slova
• biodiversity, diptera, heteroptera, host specificity, monoxenous trypanosomatids, multiple infections, nanopore sequencing, phylogeny, systematics,
• MeSH
• Diptera genetika   MeSH
• fylogeneze * MeSH
• Hemiptera parazitologie   genetika   MeSH
• koinfekce * parazitologie   MeSH
• protozoální DNA genetika   analýza   MeSH
• RNA ribozomální 18S * genetika   analýza   MeSH
• Trypanosomatina * genetika   klasifikace   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Kuba epidemiologie   MeSH
• Názvy látek
• protozoální DNA MeSH
• RNA ribozomální 18S * MeSH

Widely distributed, highly prevalent and speciose, trypanosomatid flagellates represent a convenient model to address topics such as host specificity, diversity and distribution of parasitic protists. Recent studies dealing with insect parasites of the class Kinetoplastea have been focused mainly on trypanosomatids from true bugs (Heteroptera), even though flies (Diptera, Brachycera) are also known as their frequent hosts. Phylogenetic position, host specificity and geographic distribution of trypanosomatids parasitizing dipteran hosts collected in nine countries on four continents (Bulgaria, Czech Republic, Ecuador, Ghana, Kenya, Madagascar, Mongolia, Papua New Guinea and Turkey) are presented. Spliced leader (SL) RNA gene repeats and small subunit (SSU) rRNA genes were PCR amplified from trypanosomatids infecting the gut of a total of forty fly specimens belonging to nine families. While SL RNA was mainly used for barcoding, SSU rRNA was utilized in phylogenetic analyses. Thirty-six different typing units (TUs) were revealed, of which 24 are described for the first time and represent potential new species. Multiple infections with several TUs are more common among brachyceran hosts than in true bugs, reaching one third of cases. When compared to trypanosomatids from heteropteran bugs, brachyceran flagellates are more host specific on the genus level. From seven previously recognized branches of monoxenous trypanosomatids, the Blastocrithidia and "jaculum" clades accommodate almost solely parasites of Heteroptera; two other clades (Herpetomonas and Angomonas) are formed primarily by flagellates found in dipteran hosts, with the most species-rich Leishmaniinae and the small Strigomonas and "collosoma" clades remaining promiscuous. Furthermore, two new clades of trypanosomatids from brachyceran flies emerged in this study. While flagellates from brachyceran hosts have moderate to higher host specificity, geographic distribution of at least some of them seems to be cosmopolitan. Moreover, the genus Angomonas, so far known only from South America, is present on other continents as well.

• Klíčová slova
• Diversity, Geographic distribution, Host specificity, Leishmania, Phylogeny, Trypanosomatida,
• MeSH
• Bayesova věta MeSH
• biologická evoluce MeSH
• Diptera parazitologie   MeSH
• fylogeneze * MeSH
• fylogeografie MeSH
• genetická variace * MeSH
• hostitelská specificita MeSH
• modely genetické MeSH
• protozoální DNA klasifikace   genetika   MeSH
• RNA ribozomální klasifikace   genetika   MeSH
• sekvenční analýza DNA MeSH
• Trypanosomatina klasifikace   genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Afrika MeSH
• Evropa MeSH
• Latinská Amerika MeSH
• Názvy látek
• protozoální DNA MeSH
• RNA ribozomální MeSH

Compared to their relatives, the diversity of endosymbiont-containing Trypanosomatidae remains under-investigated, with only two new species described in the past 25 years, bringing the total to six. The possible reasons for such a poor representation of this group are either their overall scarcity or susceptibility of their symbionts to antibiotics that are traditionally used for cultivation of flagellates. In this work we describe the isolation, cultivation, as well as morphological and molecular characterization of a novel endosymbiont-harboring trypanosomatid species, Kentomonas sorsogonicus sp. n. The newly erected genus Kentomonas gen. n. shares many common features with the genera Angomonas and Strigomonas, such as the presence of an extensive system of peripheral mitochondrial branches distorting the corset of subpellicular microtubules, large and loosely packed kinetoplast, and a rudimentary paraflagellar rod. Here we also propose to unite all endosymbiont-bearing trypanosomatids into the new subfamily Strigomonadinae subfam. n.

• Klíčová slova
• Kentomonas, Trypanosomatidae, bacterial endosymbionts, phylogeny.,
• MeSH
• Bayesova věta MeSH
• DNA bakterií izolace a purifikace   MeSH
• DNA virů izolace a purifikace   MeSH
• druhová specificita MeSH
• fylogeneze * MeSH
• klonování DNA MeSH
• modely genetické MeSH
• pravděpodobnostní funkce MeSH
• RNA protozoální genetika   MeSH
• RNA ribozomální 18S genetika   MeSH
• Sarcophagidae parazitologie   MeSH
• sekvenční analýza DNA MeSH
• symbióza genetika   MeSH
• Trypanosomatina klasifikace   izolace a purifikace   mikrobiologie   virologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA bakterií MeSH
• DNA virů MeSH
• RNA protozoální MeSH
• RNA ribozomální 18S MeSH

Transposable elements (TEs) have the ability to move and amplify inside the host genome, making them a pivotal source of genome plasticity. Presently, only 4 TE clades (all classified as Class I retrotransposons) have been identified in trypanosomatids. We predicted repeat content and manually curated TEs across the genomes of 57 trypanosomatids, shedding light on their proportions, diversity and dynamics. Our analysis yielded 214 TE consensus sequence models across the dataset, with abundance ranging from 0.1% to 7.2%. We found evidence of recent transposon activity in most species, with notable bursts in the Vickermania, Lafontella, Porcisia and Angomonas spp., along with Leishmania (Mundinia) chancei, L. (M.) orientalis and L. (M.) procaviensis. We confirmed that the 4 TE clades have colonized virtually all lineages of trypanosomatids, potentially playing a role in shaping their genome architecture. The effort of this work culminated in the establishment of the Trypanosomatid TE Database 1.0, a resource designed to standardize the TE annotation process that can serve as a foundation for future studies on trypanosomatid TEs.

• Klíčová slova
• CRE, INGI, SLACS, TATE, VIPER, mobilome, transposable elements, trypanosomatids,
• Publikační typ
• časopisecké články MeSH

Trypanosomatids of the genera Angomonas and Strigomonas (subfamily Strigomonadinae) have long been known to contain intracellular beta-proteobacteria, which provide them with many important nutrients such as haem, essential amino acids and vitamins. Recently, Kentomonas sorsogonicus, a divergent member of Strigomonadinae, has been described. Herein, we characterize the genome of its endosymbiont, Candidatus Kinetoplastibacterium sorsogonicusi. This genome is completely syntenic with those of other known Ca. Kinetoplastibacterium spp., but more reduced in size (~742 kb, compared with 810-833 kb, respectively). Gene losses are not concentrated in any hot-spots but are instead distributed throughout the genome. The most conspicuous loss is that of the haem-synthesis pathway. For long, removing haemin from the culture medium has been a standard procedure in cultivating trypanosomatids isolated from insects; continued growth was considered as an evidence of endosymbiont presence. However, we demonstrate that, despite bearing the endosymbiont, K. sorsogonicus cannot grow in culture without haem. Thus, the traditional test cannot be taken as a reliable criterion for the absence or presence of endosymbionts in trypanosomatid flagellates. It remains unclear why the ability to synthesize such an essential compound was lost in Ca. K. sorsogonicusi, whereas all other known bacterial endosymbionts of trypanosomatids retain them.

• Klíčová slova
• Endosymbiosis, Trypanosomatidae., genome evolution, genome reduction, haem synthesis,
• MeSH
• Betaproteobacteria účinky léků   genetika   růst a vývoj   MeSH
• biosyntetické dráhy MeSH
• fylogeneze MeSH
• genom bakteriální * MeSH
• hem metabolismus   farmakologie   MeSH
• sekvenční analýza DNA MeSH
• symbióza * MeSH
• Trypanosomatina mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• hem MeSH