Biological control ofMythimna separata [Lep.: noctuidae] in New Zealand and its bearing on biological control strategy

A survey of parasites ofAgrotis spp. was carried out in Pakistan to find promising species for trial against noctuids especiallyAgrotis spp. andMythimna separata (Walker) in New Zealand. The parasites recorded wereApanteles ruficrus Haliday,Macrocentrus collaris Spinola,Periscepsia carbonaria Panzer,Turanogonia smirnovi,Rohdain,Ctenichneumon panzeri Wesmael andAnthrax sp. All exceptAnthrax sp. were supplied to New Zealand whereA. ruficrus andM. collaris were released in large numbers.A. ruficrus became established and is giving excellent control ofM. separata resulting in enormous economic gains. Recently it has also been recovered fromAgrotis spp. This example of biological control is significant in thatM. separata has been controlled by a parasite that was previously known from it in New Zealand. The specific status of the “A. ruficrus” already present in New Zealand requires investigation.     

Tree-based delimitation of morphologically ambiguous taxa: a study of the lizard malaria parasites on the Caribbean island of Hispaniola

Malaria parasites in the genus Plasmodium have been classified primarily on the basis of differences in morphology. These single-celled organisms often lack distinguishing morphological features, and this can encumber both species delimitation and identification. Six saurian malaria parasites have been described from the Caribbean island of Hispaniola. All six infect lizards in the genus Anolis, but only two of these parasites can be distinguished using morphology. The remaining four species overlap in morphology and geography, and cannot be consistently identified using traditional methods. We compared a morphological approach with a molecular phylogenetic approach for assessing the taxonomy of these parasites. We surveyed for blood parasites from 677 Anolis lizards, representing 26 Anolis spp. from a total of 52 sites across Hispaniola. Fifty-five of these lizards were infected with Plasmodium spp., representing several new host records, but only 24 of these infections could be matched to previously described species using traditional morphological criteria. We then estimated the phylogeny of these parasites using both mitochondrial (cytb and coxI) and nuclear (EF2) genes, and included carefully selected GenBank sequences to confirm identities for certain species. Our molecular results unambiguously corroborated our morphology-based species identifications for only the two species previously judged to be morphologically distinctive. The remaining infections fell into two well-supported and reciprocally monophyletic clades, which contained the morphological variation previously reported for all four of the morphologically ambiguous species. One of these clades was identified as Plasmodium floridense and the other as Plasmodium fairchildi hispaniolae. We elevate the latter to Plasmodium hispaniolae comb. nov. because it is polyphyletic with the mainland species Plasmodium fairchildifairchildi and we contribute additional morphological and molecular characters for future species delimitation. Our phylogenetic hypotheses indicate that two currently recognised taxa, Plasmodium minasense anolisi and Plasmodium tropiduri caribbense, are not valid on Hispaniola. These results illustrate that molecular data can improve taxonomic hypotheses in Plasmodium when reliable morphological characters are lacking.     

Lochkovian (earliest Devonian) miospores from the Solimões Basin,northwestern Brazil

Eighteen core samples from PETROBRAS well 1-JD-1-AM (Jandiatuba area, Solimões Basin, northwestern Brazil) have been studied for miospores. Fifty-three species are identified. This assemblage is tentatively correlated with the Z Phylozone of the BZ Oppel Zone (Steemans, P., 1989. Palynostratigraphie de l'Eodévonien dans l'ouest de l'Europe. Professional Paper. Mémoires Explicatifs pour les Cartes Géologiques and Minéralogiques de la Belgique, 27, pp. 453.), dated late Lochkovian. This is in accordance with previous age determinations based on chitinozoans and acritarchs. Numerous representatives of the miospore genus Dictyotriletes are observed. These enable recognition of the Dictyotriletes emsiensis morphon, an informal classification unit which includes Dictyotriletes granulatus, D. emsiensis, Dictyotriletes cf. subgranifer, specimens previously misassigned to D. subgranifer and some Dictyotriletes spp. provisionally left in open nomenclature. Since all intermediate forms from typical D. granulatus to typical D. emsiensis coexist, it is currently difficult to determine the base of “E” Interval Biozone in South America as originally defined in Western Europe. It is suggested, therefore, that the base of the “Ems” Biozone defined in Brazil cannot be correlated with the base of the “E” Biozone. Instead, it should be included in an interval of uncertainty ranging from the European “Si” to “E” Biozones. Indeed, the underlying “NsZ” Biozone from South America contains species not known below the “Si” Biozone. Here, the D. emsiensis morphon Assemblage Zone is erected, which may be coeval with the “N” through “E” Biozones of Western Europe. The PISA palynoflora from the Paraná Basin is also attributed to this new biozone. However, in the absence of species observed in the “BZ” Biozone (and also in the Solimões Basin), the PISA assemblage could be somewhat older than that of well 1-JD-1-AM.     

New species and new combinations in Briquetia and Hochreutinera,and a discussion of the Briquetia generic alliance (Malvaceae)

Two new species ofBriquetia,B. sonorae andB. inermis, are described from northern Mexico, and another species that ranges from Mexico to Bolivia is transferred into the genus asB. spicata. The genus (previously known as monotypic, Paraguayan-Brazilian) is thereby enlarged from one species to four, and its geographical and morphological limits are greatly expanded. Also, a Mexican species ofAbutilon is transferred toHochreutinera, asH. amplexifolia, enlarging that genus from one to two species. On the basis of this new information, the limits of the informalBriquetia alliance are reconsidered; it is considered to include onlyBriquetia, Dirhamphis, andBatesimalva.     

Discovery of a novel species,Theileria haneyi n. sp., infective to equids,highlights exceptional genomic diversity within the genus Theileria: implications for apicomplexan parasite surveillance

A novel apicomplexan parasite was serendipitously discovered in horses at the United States – Mexico border. Phylogenetic analysis based on 18S rDNA showed the erythrocyte-infective parasite to be related to, but distinct from, Theileria spp. in Africa, the most similar taxa being Theileria spp. from waterbuck and mountain zebra. The degree of sequence variability observed at the 18S rDNA locus also suggests the likely existence of additional cryptic species. Among described species, the genome of this novel equid Theileria parasite is most similar to that of Theileria equi, also a pathogen of horses. The estimated divergence time between the new Theileria sp. and T. equi, based on genomic sequence data, is greater than 33?million years. Average protein sequence divergence between them, at 23%, is greater than that of Theileria parva and Theileria annulata proteins, which is 18%. The latter two represent highly virulent Theileria spp. of domestic cattle, as well as of African and Asian wild buffalo, respectively, which differ markedly in pathology, host cell tropism, tick vector and geographical distribution. The extent of genome-wide sequence divergence, as well as significant morphological differences, relative to T. equi justify the classification of Theileria sp. as a new taxon. Despite the overall genomic divergence, the nine member equi merozoite antigen (EMA) superfamily, previously found as a multigene family only in T. equi, is also present in the novel parasite. Practically, significant sequence divergence in antigenic loci resulted in this undescribed Theileria sp. not being detectable using currently available diagnostic tests. Discovery of this novel species infective to equids highlights exceptional diversity within the genus Theileria, a finding with serious implications for apicomplexan parasite surveillance.     

Phylogenetic relationships among monogenean gill parasites (Dactylogyridea, Ancyrocephalidae) infesting tilapiine hosts (Cichlidae): systematic and evolutionary implications

We studied the systematics of 14 species of monogenean (Ancyrocephalidae) gill parasites from West African tilapiine hosts (Cichlidae) using both morphological and genetic data. With these tools, we were able to: (i) confirm the validity of the previously described morphological parasite species and of the genus Scutogyrus; (ii) propose that some stenoxenous species (i.e., parasite species with more than one host) may be composed of sister species (e.g., Cichlidogyrus tilapiae); (iii) state that the use of the morphology of the haptoral sclerites is more suitable to infer phylogenetic relationships than the morphology of the genitalia (which seems to be more useful to resolve species-level identifications, presumably because of its faster rate of change). These results imply that: (i) the specificity of these monogenean parasites is greater than initially supposed (what were thought to be stenoxenous species may be assemblages of oïoxenous sister species); (ii) related species groups (i.e., “tilapiae,” “halli,” and “tiberianus”) have to be, as genus Scutogyrus, validated within the 54 ancyrocephalid species described from 18 species of tilapiine hosts in West Africa, (iii) the group “tilapiae,” due to its morphology and host range, have to be considered as being the most primitive; (iv) the occurrence of lateral transfers and parallel speciation processes are necessary to describe the repartition of the newly described parasite groups on the three host genera studied (Tilapia, Oreochromis, and Sarotherodon).     

Lipid synthesis in protozoan parasites: A comparison between kinetoplastids and apicomplexans

Lipid metabolism is of crucial importance for pathogens. Lipids serve as cellular building blocks, signalling molecules, energy stores, posttranslational modifiers, and pathogenesis factors. Parasites rely on a complex system of uptake and synthesis mechanisms to satisfy their lipid needs. The parameters of this system change dramatically as the parasite transits through the various stages of its life cycle. Here we discuss the tremendous recent advances that have been made in the understanding of the synthesis and uptake pathways for fatty acids and phospholipids in apicomplexan and kinetoplastid parasites, including Plasmodium, Toxoplasma, Cryptosporidium, Trypanosoma and Leishmania. Lipid synthesis differs in significant ways between parasites from both phyla and the human host. Parasites have acquired novel pathways through endosymbiosis, as in the case of the apicoplast, have dramatically reshaped substrate and product profiles, and have evolved specialized lipids to interact with or manipulate the host. These differences potentially provide opportunities for drug development. We outline the lipid pathways for key species in detail as they progress through the developmental cycle and highlight those that are of particular importance to the biology of the pathogens and/or are the most promising targets for parasite-specific treatment.     

Molecular approaches to diversity of populations of apicomplexan parasites

Apicomplexan parasites include many parasites of importance either for livestock or as causative agents of human diseases. The importance of these parasites has been recognised by the European Commission and resulted in support of the COST (Cooperation in Science and Technology) Action 857 ‘Apicomplexan Biology in the Post-Genomic Era’. In this review we discuss the current understanding in ‘Biodiversity and Population Genetics’ of the major apicomplexan parasites, namely the Eimeria spp., Cryptosporidium spp., Toxoplasma gondii, Neosporacaninum, Theileria spp. and Plasmodium spp. During the past decade molecular tools for characterizing and monitoring parasite populations have been firmly established as an integral part of field studies and intervention trials. Analyses have been conducted for most apicomplexan pathogens to describe the extent of genetic diversity, infection dynamics or population structure. The underlying key question for all parasites is to understand how genetic diversity influences epidemiology and pathogenicity and its implication in therapeutic and vaccination strategies as well as disease control. Similarities in the basic biology and disease or transmission patterns among this order of parasites promote multifaceted discussions and comparison of epidemiological approaches and methodological tools. This fosters mutual learning and has the potential for cross-fertilisation of ideas and technical approaches.     

Protalveolate phylogeny and systematics and the origins of Sporozoa and dinoflagellates (phylum Myzozoa nom. nov.)

The protozoan infrakingdom Alveolata comprises the phyla Ciliophora and Miozoa. The name Myzozoa—sucking life—is introduced here to replace Miozoa (protalveolates, dinoflagellates, Sporozoa, apicomonads) as both subphyla (Dinozoa, Apicomplexa) are commonly or ancestrally myzocytotic feeders. We studied ultrastructurally two contrasting myzocytotic flagellates: Colpodella tetrahymenae sp. n. (predatory on Tetrahymena), with an inner membrane complex like Sporozoa, and Voromonas (=Colpodella) pontica gen. et comb. nov., with discrete cortical alveoli like Dinozoa; we also sequenced 18S rDNA of both of these flagellates and of two highly divergent isolates of Oxyrrhis. Phylogenetic analysis shows early divergence between Colpodella, Voromonas and Alphamonas edax and supports the independent origin of dinoflagellates, Perkinsea and Apicomplexa (Sporozoa, Apicomonadea) from myzocytotic protalveolate flagellates. Oxyrrhis is probably a highly modified dinoflagellate, not a protalveolate with primitive chromatin and ciliary organization. The rapid basal radiation of Myzozoa is poorly resolved; the predatory colpodellids sensu stricto are probably sisters of Sporozoa. We discuss early cellular diversification of Myzozoa (=Miozoa) and revise its classification, establishing a new class Myzomonadea for Voromonas, Alphamonas and Chilovora (=Bodo) perforans gen. et comb. nov., three new peridinean subclasses (Oxyrrhia, Gonaulacoidia, Suessioidia), and five new orders: Acrocoelida for Acrocoelus; Voromonadida for Voromonas and Alphamonas; Chilovorida for Chilovora; Rastromonadida for Rastromonas and Parvilucifera; and Algovorida for Algovora, a new genus for Colpodella turpis and Colpodella pugnax sensu Simpson and Patterson. We suggest that the flattened inner membrane complex of Apicomplexa evolved in association with trichocyst loss by fusion of already flattened myzomonad cortical alveoli as an adaptation for actomyosin-based host penetration and gliding motility.     

Molecular characterization of the conoid complex in Toxoplasma reveals its conservation in all apicomplexans,including Plasmodium species

The apical complex is the instrument of invasion used by apicomplexan parasites, and the conoid is a conspicuous feature of this apparatus found throughout this phylum. The conoid, however, is believed to be heavily reduced or missing from Plasmodium species and other members of the class Aconoidasida. Relatively few conoid proteins have previously been identified, making it difficult to address how conserved this feature is throughout the phylum, and whether it is genuinely missing from some major groups. Moreover, parasites such as Plasmodium species cycle through 3 invasive forms, and there is the possibility of differential presence of the conoid between these stages. We have applied spatial proteomics and high-resolution microscopy to develop a more complete molecular inventory and understanding of the organisation of conoid-associated proteins in the model apicomplexan Toxoplasma gondii. These data revealed molecular conservation of all conoid substructures throughout Apicomplexa, including Plasmodium, and even in allied Myzozoa such as Chromera and dinoflagellates. We reporter-tagged and observed the expression and location of several conoid complex proteins in the malaria model P. berghei and revealed equivalent structures in all of its zoite forms, as well as evidence of molecular differentiation between blood-stage merozoites and the ookinetes and sporozoites of the mosquito vector. Collectively, we show that the conoid is a conserved apicomplexan element at the heart of the invasion mechanisms of these highly successful and often devastating parasites.

Proteomic characterisation of the invasion-related conoid structure in Toxoplasma reveals that this structure is ubiquitous in apicomplexan parasites, including the malaria parasite Plasmodium, where it was previously thought to be absent.     

Resolution of the long-standing controversy over the type species of the genus Pseudotypotherium (Notoungulata,Typotheria, Mesotheriidae)

Mesotheres (Notoungulata: Typotheria) are among the most common mammals found in upper Miocene to Pliocene deposits of central Argentina, including the classic type Monte Hermoso locality, which defines the Montehermosan South American Land Mammal “Age”. Nevertheless, the correct name for the mesothere species from this site has been shrouded in uncertainty for well over a century due to questions of taxonomic priority, specimen provenance, and ontogenetic changes in dental formula. Since the mesotheres from Monte Hermoso were named, three distinct species have been formally considered as the type species of the genus: (1) Pseudotypotherium bravardi; (2) “Pseudotypotherium” maendrum; and (3) Pseudotypotherium exiguum. However, none of these species is a nominal species of the Pseudotypotherium genus; all three were originally referred to Typotherium. Article 67.2 of the International Code of Zoological Nomenclature (ICZN, 1999) indicates that only species considered as nominal species are eligible to set the type; in the case of Pseudotypotherium, these include: P. pulchrum, P. carlesi, P. hystatum, and P. carhuense. We conclude that Pseudotypotherium pulchrum F. Ameghino, 1904 (holotype MACN A 10299, Museo Argentino de Ciencias Naturales “Bernardino Rivadavia”, Ameghino Collection), is the type species of the mesotheriid notoungulate genus from Monte Hermoso. According to Article 68.2, F. Ameghino fixed the type by original designation in 1904 when he described P. pulchrum and included “n. g., n. sp.”. Two of the other species previously considered species P. (= T.) bravardi and P. (= T.) exiguum are invalid as type species according to Article 70.2, since their designations overlooked the previous type fixation. The third species (M. (= T.) maendrum) represents a different mesothere genus (Mesotherium) that only occurs in younger (Pleistocene) deposits. Our analysis puts an end to a historical debate that has been ongoing for more than a century regarding the identity of this well-represented late Miocene–Pliocene mesotheriine genus (Pseudotypotherium). This study provides a solid taxonomic foundation for future studies on intraspecific and ontogenetic variation of Pseudotypotherium pulchrum.     

Isolation of a new heterolobosean amoeba from a rice field soil: Vrihiamoeba italica gen. nov., sp. nov.

A heterolobosean amoeba strain 6_5F was isolated from an Italian rice field soil. Although 18S rRNA gene sequence analysis demonstrated that the new isolate was closely related to Stachyamoeba sp. ATCC 50324, further molecular analysis and morphological observation showed distinct differences amongst the two. The 5.8S rRNA gene was successfully amplified and sequenced for strain 6_5F but not for strain ATCC 50324. Trophozoites of strain ATCC 50324 transform into flagellate forms in the late stage of incubation before encystment, while strain 6_5F do not show flagellate forms under different conditions of the flagellation test. Light and electron microscopic observation showed the structural difference of cysts of strain 6_5F from strain ATCC 50324 and also from the type strain Stachyamoeba lipophora. The results show that the strain 6_5F is distinct from Stachyamoeba spp. and we propose a new genus and species for this isolate, Vrihiamoeba italica gen. nov., sp. nov.     

Pirsonia guinardiae,gen. et spec. nov.: A parasitic flagellate on the marine diatomGuinardia flaccida with an unusual mode of food uptake

Pirsonia guinardiae gen. et spec. nov. was discovered as a parasite onGuinardia flaccida in the North Sea near List/Sylt during a bloom of this centric planktonic diatom. It is a colourless, small flagellate with an oblique cell apex and two subapically inserting flagella of different length and different orientation. The flagellates attach to a host cell and form an antapical process which pierces the diatom frustule and develops inside into a “trophosome”, consisting of a proximal digestion vacuole and distal pseudopodia which phagocytise host cytoplasm. The main body, the “auxosome”, remains outside the host cell. The trophosome persists for some time after the detachment of the auxosome or its derivatives. There are two types ofPirsonia guinardiae. Type A attaches to the valvae as well as to the girdle region, the auxosome remains flagellated and generally detaches after the feeding process to divide twice (seldom 3 times). Thick-walled (resting?) cysts are formed. Occasionally, a fusion of two sister cells has been observed. Type B attaches only to the valvae; the auxosome lacks flagella; it divides during the feeding process to give rise to a bouquet of 8 to more than 50 daughter cells which become flagellated when they detach. The taxonomical position of the flagellate is discussed. Diagnoses of genus and species are given.     

Morphological Identities of Two Different Marine Stramenopile Environmental Sequence Clades: Bicosoeca kenaiensis (Hilliard, 1971) and Cantina marsupialis (Larsen and Patterson, 1990) gen. nov., comb. nov.

Although environmental DNA surveys improve our understanding of biodiversity, interpretation of unidentified lineages is limited by the absence of associated morphological traits and living cultures. Unidentified lineages of marine stramenopiles are called “MAST clades”. Twenty‐five MAST clades have been recognized: MAST‐1 through MAST‐25; seven of these have been subsequently discarded because the sequences representing those clades were found to either (1) be chimeric or (2) affiliate within previously described taxonomic groups. Eighteen MAST clades remain without a cellular identity. Moreover, the discarded “MAST‐13” has been used in different studies to refer to two different environmental sequence clades. After establishing four cultures representing two different species of heterotrophic stramenopiles and then characterizing their morphology and molecular phylogenetic positions, we determined that the two different species represented the two different MAST‐13 clades: (1) a lorica‐bearing Bicosoeca kenaiensis and (2) a microaerophilic flagellate previously named “Cafeteria marsupialis”. Both species were previously described with only light microscopy; no cultures, ultrastructural data or DNA sequences were available from these species prior to this study. The molecular phylogenetic position of three different “C. marsupialis” isolates was not closely related to the type species of Cafeteria; therefore, we established a new genus for these isolates, Cantina gen. nov.     

Ecological factors of flowering and pollen quality in three willow species

During 1977 to 1980 the course of flowering and pollen quality were investigated inSalix cinerea, S. fragilis, andS. pentandra, the dominant woody species in a wetland area called “Mokré louky” near T?eboň, South Bohemia, Czechoslovakia. Results were compared with basíc climatological data, and decisive factors of flowering were estimated. BothS. cinerea andS. pentandra produced morphologically well developed pollen with a high degree of viability. Comparison in twelve habitats showed that the pollen ofS. fragilis was very sensitive to climatic factors, and that the marsh habitat “Mokré Louky” was hostile to the generative reproduction of this species. Optimal conditions for pollen germination in vitro were defined for all the three willows.     

Meeresalgen von Helgoland: Ergänzung

Complementary to a previous publication (Kornmann & Sahling, 1977), this investigation deals mainly with microscopic algae occurring in the rocky littoral of Helgoland island (North Sea). Based on the results obtained from cultivation experiments, the heterogeneousUlvella-complex of Dangeard has been rearranged and partly included in a new genusStromatella, andPlanophila respectively. The life history ofChlorocystis cohnii proved to be heteromorphic, the zygotes developing into a Codiolum-sporophyte. Also inHalochlorococcum marinum, some of the biflagellate swarmers give rise to Codiolum-like cells.Chlorocystis andHalochlorococcum, up to now members of the Chlorococcales, are incorporated into the new Codiolophycean order, Chlorocystidales. Three newHalochlorococcum species are described, the epiphytic“Chlorochytrium” moorei also being combined with this genus. Supplementary observations on some crustose red algae from transparent substrates are included in this study, as well as findings of some species not previously reported for the Helgoland area.     

New host,geographic records,and histopathologic studies of Angiostrongylus spp (Nematoda: Angiostrongylidae) in rodents from Argentina with updated summary of records from rodent hosts and host specificity assessment

To date, 21 species of the genus Angiostrongylus (Nematoda: Angiostrongylidae) have been reported around the world, 15 of which are parasites of rodents. In this study, new host, geographic records, and histopathologic studies of Angiostrongylus spp in sigmodontine rodents from Argentina, with an updated summary of records from rodent hosts and host specificity assessment, are provided. Records of Angiostrongylus costaricensis from Akodon montensis andAngiostrongylus morerai from six new hosts and geographical localities in Argentina are reported. The gross and histopathologic changes in the lungs of the host species due to angiostrongylosis are described. Published records of the genus Angiostrongylus from rodents and patterns of host specificity are presented. Individual Angiostrongylusspecies parasitise between one-19 different host species. The most frequent values of the specificity index (STD) were between 1-5.97. The elevated number of host species (n = 7) of A. morerai with a STD = 1.86 is a reflection of multiple systematic studies of parasites from sigmodontine rodents in the area of Cuenca del Plata, Argentina, showing that an increase in sampling effort can result in new findings. The combination of low host specificity and a wide geographic distribution of Angiostrongylus spp indicates a troubling epidemiological scenario although, as yet, no human cases have been reported.     

Herpetomonas roitmani (Fiorini et al., 1989) n. comb.: a trypanosomatid with a bacterium-like endosymbiont in the cytoplasm

The trypanosomatid previously described as Crithidia roitmani is characterized here at the ultrastructural and biochemical levels. The data indicates that the parasite belongs to the Herpetomonas genus, and we therefore suggest the flagellate to be denominated as Herpetomonas roitmani n. comb. Cladistic analysis of isoenzyme data generated by eight different enzymes showed that the parasite presented a distinct banding pattern and could be grouped with some Herpetomonas spp., but not with Crithidia spp., used as reference strains. Accordingly, when the parasites were grown for longer periods in Roitman's defined medium, expontaneous differentiation from promastigotes to opisthomastigotes (typical of the Herpetomonas genus) occurred. Transmission electron microscopy revealed the presence of bacterium-like endosymbionts in the cytoplasm of all evolutive forms of the parasite. All morphological alterations characteristic of endosymbiont-bearing trypanosomatids could be observed.     

Malladaites nov. gen. et Spinammatoceras (Hammatoceratidae,Ammonitina) de l'Aalénien de la zone subbétique,Sud de l'Espagne

The aim of this paper is to remark some morphologicaldifferences among the type-species of the genus ZurcheriaDouvillé and those species attributed to this genuf by Buckman & Brasil: the group of H. pugnaxVacek for which Schindewolf (1964) defined the genus Spinammatoceras, and the group of H. pertinaxVacek for which we propose Malladaites gen. nov.; new species of both genera are described. The stratigraphical range of these species is established in the Betic Cordillera.     

Polychaos centronucleolus n. sp. – a new terrestrial species of the genus Polychaos (Amoebozoa,Tubulinea) with nontypical nuclear structure

A new species of the “proteus-type” naked amoebae (large cells with discrete tubular pseudopodia) was isolated from tree bark sample of a birch tree in the surrounding of Kislovodsk town, Russia and named Polychaos centronucleolus n. sp. (Amoebozoa, Tubulinea). Amoebae of this species have a filamentous cell coat and a nucleus with a central compact nucleolus. This type of nucleolar organization has not been previously known for the genus Polychaos. A sequence of the 18S rRNA gene of this strain was obtained using whole genome amplification of DNA from the single amoeba cell, followed by NGS sequencing. The analysis of molecular data robustly groups this species with Polychaos annulatum within the family Hartmannellidae. Our results, together with the results of our previous studies, show that the taxonomic assignment of “proteus-type” amoebae species is becoming increasingly complex, and the taxonomic characters that can be used to classify these organisms are becoming more shadowed.