Fine Structure of the Mastigont System in Tritrichomonas foetus (Riedmüller)*

SYNOPSIS. Tritrichomonas foetus shares many fine-structural features with the previously described genera of the subfamily Trichomonadinae. These include the arrangement and structure of the kinetosomes, of most rootlet filaments, including the sigmoid filaments of kinetosome #2, as well as those of the parabasal apparatus and of the pelta-axostyle complex. On the other hand, this species, and presumably all other Tritrichomonas augusta-type flagellates, differ from Trichomonadinae in certain important details. Among the features which T. foetus does not share with Trichomonadinae are the fine structure of the costa and of the undulating membrane, as well as several organelles not found in the latter subfamily. The costal base of Trichomonadinae is replaced in T. foetus and other Tritrichomonadinae by a comb-like structure, extending between the costa and the infrakinetosomal body. The suprakinetosomal body, connected to kinetosome #2 in the region of attachment of the sigmoid filaments, and the infrakinetosomal body, which appears to contribute to the proximal marginal lamella, are organelles evidently restricted to Tritrichomonadinae. The undulating membrane consists of 2 parts. The proximal part is a fold-like differentiation of the dorsal body surface, the dorsal part of which contains the proximal marginal lamella. The distal part of the undulating membrane, with no obvious physical connection to the fold, encloses the distal marginal lamella in its ventral, and the microtubules of the recurrent flagellum in its dorsal area. The organelle of T. foetus which by its size, certain structural characteristics, and relationship with the undulating membrane and some organelles, including the paracostal granules, is analogous to the costa of Trichomonadinae and of Trichomitopsis termopsidis (subfamily Tritrichomonadinae), conforms in the structure of its periodic cross-striations to that of the parabasal filaments of the latter organisms; its origin corresponds closely to that of parabasal filament 2 of Trichomonadinae.     

Phylogeny of Trichomonads Based On Partial Sequences of Large Subunit Rrna and On Cladistic Analysis of Morphological Data

ABSTRACT. Several domains of large subunit rRNA from nine trichomonad species have been sequenced. Molecular phylogenies obtained with parsimony and distance methods demonstrate the trichomonads are a monophyletic group which branches very early in the eukaryotic tree. the topology of the trees is in general agreement with traditional views on evolutionary and systematic relationships of trichomonads. A clear dichotomy is noted between the subfamily Trichomonadinae and the subfamily Tritrichomonadinae. In the latter subfamily, a second division separates the " Tritrichomonas muris -type" species from the " Tritrichomonas augusta -type" ones. Previous evolutionary schemes in which the Monocercomonadidae were regarded as the most "primitive" and the Trichomonadidae as more "evolved" are not in agreement with our molecular data. the emergence of Monocercomonas and Hypotrichomonas at the base of the Tritrichomonas lineage suggests a secondary loss of some cytoskeletal structures, the costa and undulating membrane in these genera. This is corroborated by the early branching position of Trichomitus. which possesses a costa and an undulating membrane and has usually been placed among the Trichomonadidae on the basis of cytological characters. A cladistic analysis was applied to the available morphological characters in order to produce a hierarchical grouping of the taxa reflecting their morphological diversity. Supplementary key words. Evolution, molecular phylogeny, morphological cladistic analysis.     

Tritrichomonas mobilensis n. sp. (Zoomastigophorea: Trichomonadida) from the Bolivian Squirrel Monkey Saimiri boliviensis boliviensis1

ABSTRACT. A trichomonad flagellate, Tritrichomonas mobilensis n. sp., is described from the large intestine of the squirrel monkey, Saimiri boliviensis boliviensis. The organism has a lanceolate body 7–10.5 μm in length; a well developed undulating membrane; a stout, tubular axostyle with periaxostylar rings that terminate in a cone-shaped segment projecting from the posterior end of the cell; and a moderately wide costa. The anterior flagella are about as long as the body, and the recurrent flagellum is of the acroneme type. All its characteristics suggest that the new species belongs in the Tritrichomonas augusta type of the subfamily Tritrichomonadinae.     

The Mastigont System of Trichomonas gallinae (Rivolta) as Revealed by Electron Microscopy

SYNOPSIS. The fine structure of Trichomonas gallinae has been examined by electron microscopy and correlated with previous light microscope observations. A composite diagram of the flagellate, derived from both types of examination, is presented. Details of relationships of various mastigont organelles are documented by electron micrographs. The extent of the pelta and its connection to the capitulum of the axostyle have been determined. Four types of kinetosome rootlets have been described. One consists of superficial “filaments” radiating from each of the 9 triplet microtubules of kinetosomes #1, #2 and #3. A 2nd type of rootlet structure is represented by single comma-shaped filaments emerging clockwise from kinetosomes #1 and #3. The filament from kinetosome #1 has a periodic structure similar to that of the marginal lamella with which it is believed to connect. A 3rd type of rootlet emerges from kinetosome #2 as a sheet of about 9 filaments which traverse a sigmoid course and terminate on the inner surface of the microtubules of the pelta near the peltar-axostylar junction. The 4th set of structures consists of the costa and parabasal filaments. These structures have major periodicities of similar dimension but have readily differentiable repeating units. The costa appears to originate at the kinetosome of the recurrent flagellum, but its origin is also contiguous with that of parabasal filament 2 which has some continuity with kinetosomes #2 and #3. Parabasal filament 1, on the other hand, arises solely from or near kinetosome #2. Occasional observations of a costa and a parabasal filament in juxtaposition over a great part of their length has led to the suggestion that the parabasal filament may play a role in the development of the costa. Periodic and filamentous structures have been observed in paraxostylar and paracostal granules and in nearby cytoplasm. Their possible role in providing substance for the developing axostyle and the costa is discussed. The results are discussed in the light of available information pertaining to structure of various trichomonad species as revealed by light and electron microscopy.     

Structure of Monocercomonas sp. as Revealed by Electron Microscopy*

SYNOPSIS. Monocercomonas shares many fine-structural features with all other trichomonads. These include the basic arrangement of the kinetosomes as well as of the recurrent and 3 anterior flagella. The pelta-axostyle complex and the parabasal apparatus, i.e. the Golgi complex and the periodic filaments, also conform to the trichomonad pattern. Of interest with regard to the crucial evolutionary position of Monocercomonas, considered to represent the most primitive trichomonad type, is the fact that it has some structures in common with other Monocercomonadidae and Trichomonadinae and others in common with Devescovinidae and Tritrichomonadinae. Among the former organelles are the marginal lamella and the costal base, and among the latter, the comb-like organelle situated between the infrakinetosomal body and parabasal filament 2 as well as the infrakinetosomal body. No traces of either costa or undulating membrane have been noted, but a complex structure homologous to the marginal lamella of Hypotrichomonas and Trichomonadinae is found underlying the short anteriormost portion of the recurrent flagellum that is attached to the body surface. Observations of sections of selected division stages indicate the potential of parental kinetosomes #1 and #3 to become daughter kinetosome #2.     

Fine Structure of Trichomitus batrachorum (Perty)*

SYNOPSIS. Trichomitus batrachorum (Perty) shares with Trichomonadinae most of its fine-structural characteristics, especially those pertaining to the undulating membrane that consists of a high body fold, enclosing the loop-shaped, periodic marginal lamella, and of the recurrent flagellum applied to the fold. This flagellate has also certain structures, i.e. the costal base, comb-like organelle, and well developed marginal lamella, in common with Monocercomonas. It shares with Hypotrichomonas the costal base; large pelta; very fine fibers perpendicular to, and connecting the axostylar microtubules; structural details of the most proximal segment of the marginal lamella; and general relationships between dorsal body fold (poorly developed in Hypotrichomonas) and the recurrent flagellum. All these electronmicroscopic findings support the crucial position of Trichomitus in the evolutionary sequence Monocercomonas→Hypotrichomonas→Trichomitus→ Trichomonadinae suggested previously by lightmicroscopic observations. Further, Trichomitus shares with all Tritrichomonadinae the comb-like structure, not found either in Hypotrichomonas or Trichomonadinae; and it has in common with Tritrichomonas also the costa with Type A periodicity, being the only member of Trichomonadinae with this type of supporting organelle. It appears, therefore, that Trichomitus-type organisms represent an important link in the evolution of all Trichomonadidae from Monocercomonadidae.     

A Freeze-Fracture Electron Microscope Study of Trichomonas vaginalis Donné and Tritrichomonas foetus (Riedmüller)1

Two strains of Trichomonas vaginalis, JH162A, with low pathogenicity, and Balt 44, with high pathogenicity, as well as one highly pathogenic strain, KV-1, of Tritrichomonas foetus were studied by freeze-fracture electron microscopy. The protoplasmic faces (PFs) of the cell membranes of all three strains of both species had similar numbers of intramembranous particles (IMPs); however, the particles in the external faces (EFs) of these membranes were least abundant in Trichomonas vaginalis strain Balt 44 and most numerous in those of strain JH162A of this species. In Tritrichomonas foetus strain KV-1 the number of IMPs in the EF was close to but somewhat lower than that in the mild strain of the human urogenital trichomonad. In both species, the anterior, but not the recurrent, flagella had rosette-like formations, consisting of ～9 to 12 IMPs on both the PFs and EFs. The numbers and distribution of the rosettes appeared to vary among different flagella and in different areas of individual flagella of a single organism belonging to either species. The freeze-fracture electron micrographs provided a more complete understanding of the fine structure of undulating membranes of Trichomonadinae, as represented by Trichomonas vaginalis, and of Tritrichomonadinae (the Tritrichomonas augusta-type), as exemplified by Tritrichomonas foetus, than was gained from previous transmission and scanning electron microscope studies. Typically three longitudinal rows of IMPs on the PF of the recurrent flagellum of Trichomonas vaginalis were noted in the area of attachment of this flagellum to the undulating membrane. The functional aspects of the various structures and differences between certain organelles revealed in the two trichomonad species by the freeze-fracture method are discussed.     

Molecular phylogeny of Trichomonadidae family inferred from ITS-1, 5.8S rRNA and ITS-2 sequences

The Trichomonads have been the subject of several molecular studies that reported some discrepancies both at the lower and higher taxonomic levels. The purpose of this study was to make an extensive phylogenetic analysis of the Trichomonadidae using ITS-1/5.8S/ITS-2 sequences, to better understand its phylogeny and the usefulness of this marker. ITS-1/5.8S/ITS-2 sequences of 36 strains from 14 species belonging to Trichomonadidae and Monocercomonadidae were analysed, in which 20 were newly determined. Maximum likelihood, maximum parsimony, neighbour joining, and Bayesian phylogenetic methods were employed in order to reconstruct and compare the evolutionary history of this group. Tetratrichomonas gallinarum and four strains of Tetratrichomonas sp. isolated from bull genital organs were found closely related, confirming the classification of the latter, probably as a new species. The monophyly of Tritrichomonadinae and Trichomonadinae subfamilies were corroborated, with the exclusion of Trichomitus batrachorum from the latter since it grouped consistently with Hypotrichomonas acosta. Tritrichomonas foetus, Tritrichomonas suis and potentially also Tritrichomonas mobilensis seemed to correspond to the same species. Monocercomonas sp. and Ditrichomonas honigbergii emerged as independent lineages, with their phylogenetic positions undetermined. Neither Trichomonadidae nor Monocercomonadidae were supported as monophyletic groups. The ITS-1/5.8S/ITS-2 seems to be a reliable locus for phylogenetic studies in the Trichomonadida, mainly at lower taxonomic levels, and at least up to the family level.     

Structure of Hypotrichomonas acosta (Moskowitz) (Monocercomonadidae,Trichomonadida) as Revealed by Electron Microscopy*

SYNOPSIS. The fine structure of Hypotrichomonas acosta resembles in many respects that of Trichomonadidae, and especially of members of the sub-family Trichomonadinae which have been examined to date by electron microscopy. In addition, the flagellate has certain ultrastructural differences from the latter organisms, some of which are of phylogenetic significance. Among these, the structure of the undulating membrane and the apparently occasional presence of a fine filament which may be considered as homologous to the costa of Trichomonadidae are the most important. The undulating membrane is represented by a rather low and otherwise poorly developed dorsal cytoplasmic fold with an ill-defined distal marginal lamella; the recurrent flagellum is applied near the dorsum of the fold. In a very few preparations a relatively short filament, of a diameter falling below the resolution limits of light microscope, is seen in a position which corresponds to that of the costa of Trichomonadidae. The identity of the filament as a probable rudimentary costa is supported also by the character of its periodicity. The rare appearance of the rudimentary costa among hundreds of sections may be explained either by its minute dimensions or by its absence from many hypotrichomonads. Other structures recorded for the first time in trichomonads are: the fine filamentous connections of the axostylar microtubules; the branching of parabasal filament 2; and the unusually organized, perhaps helical, polysomes, which are found in addition to the ribosomal complexes associated with the endoplasmic reticulum and commonly found in trichomonads. A detailed analysis of interconnections among various mastigont structures is presented and several kinds of cytoplasmic inclusions are described. H. acosta is of interest in the study of the nuclear envelope and presence of nuclear pores, which are numerous and conspicuous in this flagellate. The fine structure of the hypotrichomonad is discussed in relation to that of other trichomonads and in some instances to that of other protozoa.     

Structure of Pentatrichomonas hominis (Davaine) as Revealed by Electron Microscopy*

SYNOPSIS. Fine structure of Pentatrichomonas hominis is described in the light of previous light microscopic findings. The relationships among kinetosomes #1-#4 and R are like those previously reported orhomonas gallinae, and the same is true of the rootlet filaments associated with the several kinetosomes. The kinetosome (I) of the independent flagellum is situated just behind the reflection of the sigmoid filaments of kinetosome #2 onto the pelta and parallels these filaments for a considerable distance. The peltaraxostylar junction consists of 3 layers: the capitulum of the axostyle (outer, the pelta (intermediate, and the sigmoid rootlets of kineto some #2 (inner). The pelta overlaps the axostylar capitulum to a variable extent. The parabasal body consists of elongate and flattened cisternae of smooth endoplasmic reticulum surrounded by numerous small vesicles. There are 2 typically cross-striated parabasal filaments, filament 2 probably contributing most, if not all, the material to the slender, periodic organelle that underlies the parabasal body and usually does not extend far beyond the posterior end of the nucleus. The periodic costa is paralleled by paracostal granules, but there are few, if any, paraxostylar granules. The ultrastructure of the costa appears to be a network of flattened hexagons, with a single fibril projecting thru each of the hexagonal areas. The major cross-striations are made up largely of densely-stained filaments which are occasionally cut in cross section. The undulating membrane consists of a cytoplasmic fold extending from the dorsal surface of the organism and of the attached part of the recurrent flagellum, which is closely applied to the fold. The segment of the membrane dorsal to the flagellum, presumably the “accessory filament,” contains the marginal lamella, a membrane folded upon itself and with periodicity virtually indistinguishable from that of the rootlet filament of kinetosome #1.     

Molecular phylogeny of parabasalids based on small subunit rRNA sequences, with emphasis on the Trichomonadinae subfamily

We determined small subunit ribosomal DNA sequences from three parabasalid species, Trichomitus batrachorum strain R105, Tetratrichomonas gallinarum, and Pentatrichomonas hominis belonging to the Trichomonadinae subfamily. Unrooted molecular phylogenetic trees inferred by distance, parsimony, and likelihood methods reveal four discrete clades among the parabasalids. The Trichomonadinae form a robust monophyletic group. Within this subfamily T. gallinarum is closely related to Trichomonas species as supported by morphological data, with P. hominis and Pseudotrypanosoma giganteum occupying basal positions. Our analysis does not place T. batrachorum within the Trichomonadinae. Trichomitus batrachorum (strains R105 and BUB) and Hypotrichomonas acosta form a well-separated cluster, suggesting the genus Trichomitus is polyphyletic. The emergence of T. batrachorum precedes the Trichomonadinae-Tritrichomonadinae dichotomy, emphasizing its pivotal evolutionary position among the Trichomonadidae. A third cluster unites the Devescovinidae and the Calonymphidae. The fourth clade contains the three hypermastigid sequences from the genus Trichonympha, which exhibit the earliest emergence among the parabasalids. The addition of these three new parabasalid species did not however resolve ambiguities regarding the relative branching order of the parabasalid clades. The phylogenetic positions of Tritrichomonas faetus, Monocercomonas sp., Dientamoeba fragilis, and the unidentified Reticulitermes flavipes gut symbiont 1 remain unclear.     

Determination of internal transcribed spacer regions (ITS) in Trichomonas vaginalis isolates and differentiation among Trichomonas species

The nucleotide sequence of the 5.8S rRNA gene and the flanked internal transcribed spacer (ITS) regions of six Trichomonas vaginalis isolates with different metronidazole sensitivity and geographic origin were genotyped. A multiple sequence alignment was performed with different sequences of other isolates available at the GenBank/EMBL/DDBJ databases, which revealed 5 different sequence patterns. Although a stable mutation in position 66 of the ITS1 (C66T) was observed in 26% (9/34) of the T. vaginalis sequences analyzed, there was 99.7% ITS nucleotide sequence identity among isolates for this sequence. The nucleotide sequence variation among other species of the genus Trichomonas ranged from 3.4% to 9.1%. Surprisingly, the % identity between T. vaginalis and Pentatrichomonas hominis was ~ 83%. There was > 40% divergence in the ITS sequence between T. vaginalis and Tritrichomonas spp., including Tritrichomonas augusta, Tritrichomonas muris, and Tritrichomonas nonconforma and with Tetratrichomonas prowazeki. Dendrograms grouped the trichomonadid sequences in robust clades according to their genera. The absence of nucleotide divergence in the hypervariable ITS regions between T. vaginalis isolates suggests the early divergence of the parasite. Importantly, these data show this ITS1-5.8S rRNA-ITS2 region suitable for inter-species differentiation.     

Tritrichomonas mobilensis n. sp. (Zoomastigophorea: Trichomonadida) from the Bolivian squirrel monkey Saimiri boliviensis boliviensis

A trichomonad flagellate, Tritrichomonas mobilensis n. sp., is described from the large intestine of the squirrel monkey, Saimiri boliviensis boliviensis. The organism has a lanceolate body 7-10.5 micrometers in length; a well developed undulating membrane; a stout, tubular axostyle with periaxostylar rings that terminate in a cone-shaped segment projecting from the posterior end of the cell; and a moderately wide costa. The anterior flagella are about as long as the body, and the recurrent flagellum is of the acroneme type. All its characteristics suggest that the new species belongs in the Tritrichomonas augusta type of the subfamily Tritrichomonadinae.     

Pseudocrossidium perpapillosum (Pottiaceae) a new species from South America

Abstract

Pseudocrossidium perpapillosum M.J.Cano & J.A.Jiménez is described and illustrated as a new South American species from Argentina and Chile. It differs from other species of Pseudocrossidium R.S.Williams by the costa excurrent in an awn, elliptical in cross-section, with 4–5(6) guide cells, marginal laminal cells differentiated with rounded, thin-walled and papillose cells, paracostal distal cells with 1–2(4) bi- to quadrifurcate, long papillae, and perichaetial leaves differentiated.     

A freeze-fracture electron microscope study of Trichomonas vaginalis Donné and Tritrichomonas foetus (Riedmüller)

Two strains of Trichomonas vaginalis, JH162A , with low pathogenicity, and Balt 44, with high pathogenicity, as well as one highly pathogenic strain, KV-1, of Tritrichomonas foetus were studied by freeze-fracture electron microscopy. The protoplasmic faces ( PFs ) of the cell membranes of all three strains of both species had similar numbers of intramembranous particles (IMPs); however, the particles in the external faces (EFs) of these membranes were least abundant in Trichomonas vaginalis strain Balt 44 and most numerous in those of strain JH162A of this species. In Tritrichomonas foetus strain KV-1 the number of IMPs in the EF was close to but somewhat lower than that in the mild strain of the human urogenital trichomonad . In both species, the anterior, but not the recurrent, flagella had rosette-like formations, consisting of approximately 9 to 12 IMPs on both the PFs and EFs. The numbers and distribution of the rosettes appeared to vary among different flagella and in different areas of individual flagella of a single organism belonging to either species. The freeze-fracture electron micrographs provided a more complete understanding of the fine structure of undulating membranes of Trichomonadinae , as represented by Trichomonas vaginalis, and of Tritrichomonadinae (the Tritrichomonas augusta -type), as exemplified by Tritrichomonas foetus, than was gained from previous transmission and scanning electron microscope studies. Typically three longitudinal rows of IMPs on the PF of the recurrent flagellum of Trichomonas vaginalis were noted in the area of attachment of this flagellum to the undulating membrane. The functional aspects of the various structures and differences between certain organelles revealed in the two trichomonad species by the freeze-fracture method are discussed.     

The structural organization of the pathogenic protozoan Tritrichomonas foetus as seen in replicas of quick frozen,freeze-fractured and deep etched cells

Summary— The quick-freezing and freeze-etching technique was used to analyse the cytoskeleton of Tritrichomonas foetus, a pathogenic protozoan of the urogenital tract of cattle. The cytoplasm presented a network of filamentous structures interacting with each other, with the surface of the hydrogenosomes and the nuclear membrane. Two nm wide filamentous structures were found in the luminal space of the Golgi complex, connecting the two faces of each cisterna. The microtubules of the pelta-axostyle system were connected by bridges 30–40 nm long and 10 nm wide, regularly spaced with an interval of 25 nm. The costa is a structure formed by a complex array of filaments and globous structures. It seems to be connected to the recurrent flagellum through a complex network formed by 15 and 10 nm wide filaments which emerge from the peripheral region of the costa and penetrate into the surface projections of the protozoan body to which the recurrent flagellum is attached. Other filaments were seen connecting the surface of these projections with the surface of the flagellum.     

BIOCHEMICAL CYTOLOGY OF TRICHOMONAD FLAGELLATES : I. Subcellular Localization of Hydrolases, Dehydrogenases, and Catalase in Tritrichomonas foetus

To determine the localization of several enzymes in Tritrichomonas foetus, the axenic KV-1 strain was grown in Diamond's medium with bovine serum, homogenized in 0.25 M sucrose, and subjected to analytical differential and isopycnic centrifugation. The fractions were assayed for their enzymatic composition and examined electron microscopically. NADH and NADPH dehydrogenases, about 90% of the catalase, and two hydrolases, α-galactosidase and manganese-activated β-galactosidase I are in the nonsedimentable part of the cytoplasm. α-Glycerophosphate and malate dehydrogenases are associated with a large particle, whose equilibrium density in sucrose gradients is 1.24. This particle corresponds to that population of the paracostal and paraxostylar granules which, having a uniform granular matrix surrounded by a single membrane, resemble microbodies from other organisms. The small sedimentable portion of catalase (about 10% of the total activity) is not associated with these granules and equilibrates at density 1.22. The nature of the subcellular entity carrying catalase could not be ascertained. Hydrolases with a pH optimum around 6–6.5 (protease, β-N-acetylglucosaminidase, β-N-acetylgalactosaminidase, and cation-independent β-galactosidase II), as well as a large part of acid phosphatase, are associated with a population of large particles which equilibrate at densities from 1.15 to 1.20. The hydrolases in these granules lose their structure-bound latency easily after freezing and thawing. These particles correspond to another population of the paracostal and paraxostylar granules which have varied shape and inhomogeneous content with frequent myelin figures, indicating a digestive function. The rest of the phosphatase and most of the acid β-glucuronidase activity are in a smaller granule fraction with an equilibrium density around 1.18. The latency of these enzymes is quite resistant to freezing and thawing. This particle population consists of smaller, very often flattened vesicles and granules, many of which are clearly fragments of the prominent Golgi apparatus of the cell.     

Morphogenesis of the hydrogenosome: An ultrastructural study

Summary— The morphogenesis of hydrogenosomes in several trichomonad species (Tritrichomonas foetus, Trichomonas vaginalis, Tritrichomonas suis, Trichomonas gallinae, Tritrichomonas augusta and Monocercomonas sp) was investigated by transmission electron microscopy of thin sections and freeze-fracture replicas of whole cells or the isolated organelle. Close proximity, and even continuity, between endoplasmic reticulum and hydrogenosomes was observed. Structures were seen connecting hydrogenosomes to each other and to cytoplasmic structures. Morphological evidence is presented showing that in all the trichomonads here studied, hydrogenosomes, like mitochondria, may divide by two distinct processes: segmentation and partition. In the segmentation process, the hydrogenosome grows, becoming enlongated with the appearance of a constriction in the central portion. Microfibrillar structures appear to help the furrowing process, ending with a total fission of the organelle. In the partition process, the division begins by an invagination of the inner hydrogenosome membrane, forming a transversal septum, separating the organelle matrix into two compartments. We suggest that myelin-like structures seen either in close contact with or in the vicinity of the hydrogenosomes may be a source of membrane lipids for hydrogenosome growth.     

Evolution and Diversity of Amitochondrial Zooflagellates

ABSTRACT. The amitochondrial flagellates comprise the Archamoebae, the Metamonada (orders: Retortamonadida, Diplomonadida, Oxymonadida) and the Parabasala. Molecular rDNA sequence comparison has shown that the diplomonads are very ancient and placed at the base of the tree, but the position of the parabasalids before or after the Euglenozoa and other mitochondriate protists is not definitively determined and such molecular data are required to place the other groups. Common cytological features such as one basal body and a basal body-nucleus connector show that the mastigamoebids or Archamoebae are an evolutionary lineage. The metamonad grouping is heterogenous; the three orders have in common two pairs of basal bodies, no Golgi and could be poly- or paraphyletic. The Parabasala is an evolutionary lineage with a large set of homologous cytological characters. A biochemical study of the cytoskeletal structures such as costa, parabasal fibre, preaxostylar fibre and undulating membrane in trichomonads reveals a large diversity in the protein composition among the genera that fits with the large distance between the taxa estimated by rDNA sequencing. Molecular comparison in seven tnchomonad genera indicates that the Trichomonadinae and Tritrichomonadinae form two divergent branches, the genera Hypotrichomonas and Monocercomonas are now incorporated in the tritrichomonad branch, and Trichomitus is at the bifurcation of the two branches. This scheme is different from the classic tree but not incompatible with it.     

Morphology of <Emphasis Type="Italic">Mastigamoeba aspera</Emphasis> schulze, 1875 (Archamoebae,Pelobiontida)

The morphology of Mastigamoeba aspera, a typical species of the genus Mastigamoeba Schulze, 1875, was studied at the optical and electron microscopy level. During movement, M. aspera has an oval or pyriformic shape, with the motile flagella being located at the anterior end of mononuclear forms. In the process of movement, the mastigamoeba surface forms numerous conical or finger-shaped hyaline pseudopodia, whereas thel caudal cell end is usually transformed into a bulboid uroid. In M. aspera micropopulations, there are noted both mononuclear cells with flagella and multinuclear flagella-free individuals. The M. aspera plasma membrane has at its outer surface a hypertrophied glycocalix layer inhabited by numerous rod-shaped bacteria-ectobionts. The M. aspera nucleus is of vesicular type, with a large central spherical nucleolus. The flagellar apparatus is closely connected morphologically with the M. aspera nucleus. The basal flagella part is represented by a single kinetosome, from which radial microtubules and a lateral rootlet pass out into the cytoplasm. At the base of the kinetosome, there is located a compact center of organization of microtubules (COMT), in which there are immersed bases of the nuclear cone microtubules participating in formation of karyomastigont. The structure of the flagella axoneme corresponds to the formula 9(2)+2. The main volume of the M. aspera cytoplasm is occupied with digestive vacuoles. In addition, the cells contain numerous light-reflecting granules, as well as glycogen granules. Mitochondria, dictyosomes of the Golgi apparatus, and microbodies in the M. aspera cell cytoplasm are not revealed.