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Morphology of Stenophora pachyiuli sp.n. A -young trophozoite; B -trophozoite; C -gamonts; D -protomerite, SEM; E -oval nucleus with an eccentric round nucleolus; F -gamonts with contracted anterior part of deutomerite. Abbreviations: d -deutomerite; N -nucleus; n -nucleolus; p -protomerite. Scale bars: A -20 µm; B -50 µm; C -100 µm; D -10 µm; E -20 µm; F -20 µm.
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... epicytic folds that start from the apical pole on the protomerite papilla (Fig. 3D). Gregarines demonstrated gliding motility, the ability to retract protomerite into deutomerite (Fig. 3E), and to recurve the front end of the cells (Fig. 3F) obdeltoid in shape up to 451.2 µm in length (av. 337 ± 16.4 µm) and 100.8 µm in width (av. 70.7 ± 3.7 µm) (Fig. 4A, B, C). All forms had a domeshaped protomerite with a small papilla at its apex. The cell surface was organized in the epicytic folds that start from the protomerite papilla (Fig. 4D). An oval nucleus with an eccentric round nucleolus might be located in any part of deutomerite (Fig. 4B, C, E). Gregarines demonstrated gliding motility ...
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... 3E), and to recurve the front end of the cells (Fig. 3F) obdeltoid in shape up to 451.2 µm in length (av. 337 ± 16.4 µm) and 100.8 µm in width (av. 70.7 ± 3.7 µm) (Fig. 4A, B, C). All forms had a domeshaped protomerite with a small papilla at its apex. The cell surface was organized in the epicytic folds that start from the protomerite papilla (Fig. 4D). An oval nucleus with an eccentric round nucleolus might be located in any part of deutomerite (Fig. 4B, C, E). Gregarines demonstrated gliding motility and the ability to contract the anterior part of deutomerite to move the protomerite (Fig. 4F). Gametocysts and oocysts are unknown. DNA SEQUENCE. GenBank OP423031. TYPE LOCALITY. ...
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... 337 ± 16.4 µm) and 100.8 µm in width (av. 70.7 ± 3.7 µm) (Fig. 4A, B, C). All forms had a domeshaped protomerite with a small papilla at its apex. The cell surface was organized in the epicytic folds that start from the protomerite papilla (Fig. 4D). An oval nucleus with an eccentric round nucleolus might be located in any part of deutomerite (Fig. 4B, C, E). Gregarines demonstrated gliding motility and the ability to contract the anterior part of deutomerite to move the protomerite (Fig. 4F). Gametocysts and oocysts are unknown. DNA SEQUENCE. GenBank OP423031. TYPE LOCALITY. Nickel, Adygea Republic, Russia (44°10′38.2″N ...
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... apex. The cell surface was organized in the epicytic folds that start from the protomerite papilla (Fig. 4D). An oval nucleus with an eccentric round nucleolus might be located in any part of deutomerite (Fig. 4B, C, E). Gregarines demonstrated gliding motility and the ability to contract the anterior part of deutomerite to move the protomerite (Fig. 4F). Gametocysts and oocysts are unknown. DNA SEQUENCE. GenBank OP423031. TYPE LOCALITY. Nickel, Adygea Republic, Russia (44°10′38.2″N ...
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... of parasite cells and host material fixed in ethanol have been deposited in the collection of The Center for Parasitology IPEE RAS; extracted DNA used for obtaining of rDNA sequences deposited in the collection of the Department of evolutionary biochemistry, Belozersky Institute for Physico-Chemical Biology, Lomonosov Moscow State University; Fig. 4 (this publication) shows some of the ...
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... Gametocyst protoplasm samples provide a more pragmatic source of genomic DNA than do samples from other life cycle stages. Trophozoites and gamonts have been used successfully by many authors (Leander et al., 2003;Rueckert and Leander, 2008;Clopton, 2009;Simdyanov et al., 2015;Diakin et al., 2016;Miroliubova et al., 2023), but these ontogenetic stages are subject to contamination, require significant effort to obtain material for a single pooled extraction, and produce mixed species extractions when morphologically similar species infecting the same host are accidentally pooled. The folded nature of the pellicle makes it difficult to clean all of the host cells and luminal debris from individual specimens, providing a problematic source of host and other non-target DNA. ...
... All new sequences were obtained using the primers and methods detailed above. Sequences representing the Stenophoroidea and Stylocephaloidea were included as outgroups to root the tree based on existing phylogenic estimates for the group (Clopton, 2009;Miroliubova et al., 2023). ...
... The phylogeny demonstrates the polyphyletic nature of Gregarina, which comprises 4 sister groups in the coleopteran-exploiting clade and 1 orthopteran-exploiting clade within Gregarinoidea. Earlier analyses (Clopton, 2009;Clopton and Clopton, 2022;Miroliubova et al., 2023) recovered similar topologies indicating the polyphyletic nature of Gregarina, and some taxonomic corrections have already been made. Amoebogregarina was removed from Gregarina by Kula and Clopton (1999). ...
Gregarina lutescens n. sp. is described from the alimentary canal of the harlequin ladybird or multicolored Asian lady beetle, Harmonia axyridis (Coleoptera: Coccinellidae) collected from prairie fleabane, Erigeron strigosus, at Peru State College, Peru, Nemaha Co., Nebraska. Our specimens differ from all 11 known species of Gregarina infecting coccinellid beetles worldwide by differences in size and relative shape, color, and association structure. Gregarina lutescens n. sp. is smaller than 7 known species infecting coccinellid beetles but larger than the other 4 known species based on confidence interval exclusion of means. Our specimens are unique among known species of interest in their quince-yellow cytoplasm and precocious but ephemeral serial associations of up to 5 satellites. Nucleotide sequence (18S) phylogenetic analyses place the new species basal to a member of an internal clade of Gregarina that comprises gregarines parasitizing chrysomelid beetles. Phylogenetically, the analysis recovered 3 major lineages within the gregarines, representing the superfamilies Gregarinoidea, Stenophoroidea, and Stylocephaloidea and indicating the propensity of gregarines to track host lineages and environments through evolutionary time. These findings confirm the polyphyletic nature of Gregarina, which currently comprises over 300 described species, only a handful of which have documented genetic sequences suitable for phylogenetic analysis. Recollection, redescription, and molecular clarification of gregarine species infecting coccinellids would likely result in identification of a unique clade that would be an excellent system for studying the effect of intraguild host competition on parasite diversification and community structure. Ecologically, patterns of prevalence in this study indicate that G. lutescens reproduces primarily in larval hosts but depends on infections in adult beetles to overwinter, reflecting the differential vagility and frost tolerance of larval and adult host life cycle stages.
Gregarines from the families Dactylophoridae and Trichorhynchidae parasitize exclusively centipedes and have a distinct morphology among other terrestrial eugregarines, but their evolutionary relationships have not yet been studied with molecular methods. Here we obtain rDNA operon sequences for the dactylophorids and trichorhynchids. We describe a new species Trichorhynchus efeykini sp. n. from a scutigeromorph Thereuopoda longicornis from Vietnam. Phylogenetic analyses with combined SSU, 5.8S and LSU rDNA dataset support the previously proposed separation of Trichorhynchus to the Trichorhynchidae based on morphology and recover the dactylophorids and trichorhynchids as sister groups in a monophyletic clade. This clade appears sister to the clade of the Actinocephaloidea and Stylocephaloidea, and represents a new major lineage of terrestrial gregarines that we designate as a new superfamily Dactylophoroidea.
