Morphology, morphogenesis and molecular phylogeny of a new soil ciliate, Lamtostyla paravitiphila nov. spec. (Ciliophora, Hypotrichia).

The morphology and morphogenesis of Lamtostyla paravitiphila nov. spec., a novel soil hypotrichous ciliate collected from eastern China, were investigated based on live observations and protargol-stained specimens. The new species is morphologically characterized as follows: seven to twelve macronuclear nodules, cortical granules absent, 19-26 adoral membranelles, three or four frontoventral cirri, the amphisiellid median cirral row extends to about mid-body and composed of 12-18 cirri, two or three transverse cirri, 27-39 left and 30-41 right marginal cirri, three almost bipolar dorsal kineties. Morphogenetically, it is characterized by the initial formation of six frontal-ventral-transverse cirral anlagen as primary primordia. Notably, the amphisiellid median cirral row and the posterior frontoventral cirrus (or cirri) contribute to the development of the frontal-ventral-transverse cirral anlagen, while the buccal cirrus may not participate in this process. Phylogenetic analyses based on small subunit ribosomal DNA sequence data indicate that the Lamtostyla species with available molecular data do not form a monophyletic group.

Morphological, ontogenetic, and phylogenetic research of a poorly known non-dorsomarginalian ciliate: Erniella filiformis Foissner, 1987 (Protista, Ciliophora, Hypotrichia).

Erniella Foissner, 1987 is a poorly known genus with its available information limited solely to morphological characteristics. In this study, the type species E. filiformis, isolated from highly saline water in China, was investigated for morphology, morphogenesis, and phylogeny based on small subunit ribosomal DNA sequences. The main characteristic events during morphogenesis in Erniella filiformis are as follows: (1) in the proter, only the posterior part of the parental adoral zone is renewed and undulating membranes anlage is formed from the dedifferentiation of the old structure; (2) the oral primordium is formed intrakinetally in the opisthe; (3) frontoventral-transverse cirral anlagen II-IV develop in the primary mode, anlage V develops intrakinetally in the proter; and (4) the right frontoventral row is formed by a single anlage. The phylogenetic analyses indicate that Erniella forms a monophyletic group with Bistichella, Keronopsis, Lamtostyla, Orthoamphisiella, Parabistichella, Paraholosticha, and Uroleptoides.

Morphology, morphogenesis and molecular phylogeny of Lamtostyla granulifera sinensis subsp. nov. (Ciliophora, Hypotrichia) from a wetland in China.

The morphology, morphogenesis and molecular phylogeny of a hypotrichous ciliate, Lamtostyla granulifera sinensis subsp. nov., isolated from northern China, were investigated. This population appeared highly similar in morphology to L. granulifera Foissner, 1997. However, on detailed investigation some non-overlapping features were identified, i.e., the body shape and the arrangement of the cortical granules. These differences suggested the separation at subspecies level. Furthermore, the morphogenesis of the new subspecies is described, which is characterized by: (1) the posterior part of the parental adoral zone of membranelles is renewed; (2) the amphisiellid median cirral row is formed from two anlagen; and (3) the frontoventral-transverse cirral anlagen II to VI generate one transverse cirrus each. Phylogenetic analyses based on SSU rDNA sequence data show that Lamtostyla species are scattered in different clades. The monophyly of the genus Lamtostyla is also rejected by the AU test in this study.

Diophrys pauciciliata n. sp. (Ciliophora, Euplotida), a new marine spirotrich ciliate with highly static morphometrics.

A new marine ciliate was discovered from intertidal mudflat from South Korea. Based on the observation of living cells, protargol-impregnated specimens and SEM, we describe it as new species belonging to the genus Diophrys Dujardin, 1841, namely Diophrys pauciciliata n. sp. Its nuclear small subunit ribosomal RNA gene also supports the taxonomic assignment. Diophrys pauciciliata n. sp. is one of the smallest species in the genus. Besides, the bipartite adoral zone and oligomerized dorsal bristles easily discriminate the new species from other congeners. Interestingly, D. pauciciliata n. sp. showed highly static states in all morphometrics (<8.5 % of coefficient of variation) and in particular, the number of frontal adoral membranelles and the counts of dorsal ciliature (e.g., number of dorsal kineties and numbers of dikinetids within each kinety) are invariable. Considering morphometric data of previous studies in the genus Diophrys, the narrow range of character states is unexpected because cell length is usually positively correlated with the numbers of adoral membranelles and dorsal dikinetids.

Morphology and morphogenesis of the hypotrich ciliate Parentocirrus hortualis Voß, 1997, with notes on the phylogeny of Parentocirrus (Ciliophora, Hypotrichia).

The morphology, morphogenesis and molecular phylogeny of the hypotrich ciliate Parentocirrus hortualis, isolated from soil in northwestern China, was studied using live observation and protargol staining. The Chinese population closely matches the German type population although some features such as the number of macronuclear nodules (4-8 vs 5-16) and transverse cirri (4 or 5 vs 2-4) differ considerably. These differences can probably be explained, at least in part, by environmental adaptability. We also compared the ontogenetic processes in the two populations and did not find any significant differences. Phylogenetic analyses based on SSU rDNA gene sequence data showed that Parentocirrus hortualis clusters with Gastrostyla steinii and Parentocirrus sp., which together form a clade with another species of Gastrostyla within the Stylonychinae. These findings suggest that Parentocirrus and Gastrostyla are closely related.

Morphology and molecular phylogeny of Keronopsis koreana n. sp. (Ciliophora, Hypotricha), a new soil species from South Korea.

The morphology and molecular phylogeny of a new soil ciliate, Keronopsis koreana n. sp., which was discovered in soil from Jindo-gun, South Korea, were investigated. The new species is characterized by the following morphological features: cell more or less ellipsoid or ovoid, both ends broadly rounded; cortical granules irregularly distributed, colorless, 1 µm across; 35-54 adoral membranelles; 12-23 cirri in frontal corona; two frontal and two frontoventral cirral rows; two or three buccal cirri; 23-38 left and 22-37 right marginal cirri; three bipolar dorsal kineties; and two macronuclear nodules and three to five micronuclei. Phylogenetic analyses show that Keronopsis koreana n. sp. is placed within a clade containing K. helluo (type species of Keronopsis) and species belonging to Paraholosticha, Uroleptoides, Parabistichella, Orthoamphisiella, Lamtostyla, and Bistichella in non-dorsomarginalian hypotrichs. Also, the sequences belonging to the Keronopsidae did not cluster together. In addition, a new combination is provided: Parabistichella tasmaniensis (Blatterer and Foissner, 1988) n. comb. (original combination: Keronopsis tasmaniensis).

Morphology, morphogenesis and phylogeny of a new soil ciliate, Bistichella sinensis n. sp., and morphology of two oxytrichids (Ciliophora, Hypotrichia).

The living morphology, infraciliature, morphogenesis and phylogenetic position of a new soil ciliate, Bistichella sinensis n. sp. collected from Northwest of China, were investigated. The new species is characterized by an elongate-oval body, two macronuclear nodules, colourless cortical granules, three frontal and three or four buccal cirri, three frontal rows with 7-10 cirri in total, and usually-two frontoventral rows with the left one usually terminating at 90% down length of body. The main morphogenetic features of the novel species are as follows: (1) the posterior part of the parental adoral zone is renewed; (2) frontal-ventral cirral anlagen III to V each forms a frontal row, and anlagen VI to n each produces a frontoventral row; (3) marginal rows and dorsal kineties develop intrakinetally. Furthermore, the phylogenetic analyses based on small subunit ribosomal DNA reveal close relationship between Bistichella sinensis n. sp., Parabistichella, Uroleptoides, Lamtostyla, Keronopsis, Paraholosticha and Orthoamphisiella. The in vivo morphology and the infraciliature of the Chinese populations of Monomicrocaryon euglenivorum fimbricirratum and Laurentiella strenua are basically identical to previous descriptions. Improved diagnoses for M. euglenivorum and its two subspecies, as well as a redescription of a Chinese population of L. strenua are supplied.

Insights into the phylogeny of the family Deviatidae (Protozoa, Ciliophora, Hypotrichia) based on multi-gene, morphological and ontogenetic information, with the establishment of a new species Deviata multilineae n. sp.

Hitherto, the phylogeny of ciliated protists, an important group of model organisms in many fields, has been mainly based on a single marker gene (SSU rDNA, nuclear small subunit ribosomal RNA gene). However, there is increasing evidence showing this is insufficient to provide robust phylogenies and has resulted in confusing systematics in many ciliates groups. Among these, the phylogenies within family Deviatidae (Spirotrichea, Hypotrichia) are ambiguous due to the dependence on SSU rDNA and undersampling. Here, we provide eight new sequences and conduct phylogenetic analyses based on both multi-gene and single-gene to clarify evolutionary relationships among all deviatids for which gene sequence are available. The results reveal that: (1) the monophyly of Deviatidae is well-supported by both single-gene and concatenated data; (2) the presence of fine cirri and relatively wide spacing of these cirri within all rows are plesiomorphies of Deviatidae; (3) Pseudosincirra longicirrata is closely related to Deviata rositae, which is supported by their shared possession of dorsomarginal kineties; (4) phylogenetic analyses and approximately unbiased test based on multi-gene support a close relationship among taxa lacking dorsomarginal kineties (D. parabacilliformis, D. multilineae nov. spec., D. abbrevescens, D. brasiliensis and Perisincirra paucicirrata); (5) Deviatidae shows a close relationship with Dorsomarginalia and Strongylidium-Hemiamphisiella-Pseudouroleptus assemblage, suggesting the presence/absence of dorsomarginal kineties is phylogenetically informative in this family and presence of them may be a plesiomorphy. Based on the morphological, morphogenetic and phylogenetic data, the evolutionary relationships within Deviatidae are hypothesized, and a new ciliate, Deviata multilineae nov. spec., collected from China, is investigated.

Morphology, Morphogenesis and Molecular Phylogeny of a New Soil Ciliate, Holostichides eastensis nov. spec. (Ciliophora, Hypotrichia).

A new bakuellid ciliate, Holostichides eastensis nov. spec. isolated from China, is investigated in terms of its morphology, ontogenesis, and molecular phylogenesis. It is characterized by size 150-180 µm × 45-60 µm in vivo; 27-46 macronuclear nodules; cortical granules dark brown, spherical, about 0.5 µm across, clustered in groups and then arranged longitudinally; three frontal cirri with one to four smaller cirri below the middle one, a buccal cirrus, nine to 15 frontoterminal cirri, and midventral complex composed of five to eight cirral pairs and two long midventral rows. The main events during binary fission are as follows: (1) in the proter, the undulating membrane anlage is formed from both the dedifferentiation of old undulating membranes and the basal bodies developing de novo, and the proximal part of the parental adoral zone of membranelles is renewed by a field of basal bodies that originates de novo; (2) in the opisthe, the oral primordium is formed intrakinetally; (3) besides the middle frontal cirrus and buccal cirrus, the FVT-anlagen II produces the extra one to four cirri. Phylogenetic analyses based on SSU rDNA sequence data suggest a close relationship between Holostichides eastensis nov. spec. and its congeners, indicating that the genus Holostichides is monophyletic.

Morphology, ontogeny and molecular phylogeny of a new urostylid ciliate, Bakuella (Pseudobakuella) guangdongica n. sp. (Ciliophora, Hypotrichia) from southern China.

The morphology, morphogenesis, and molecular phylogeny of Bakuella (Pseudobakuella) guangdongica n. sp., isolated from southern China, were investigated. The new species is characterized by a body length of 150-225 µm in vivo; 35-42 adoral membranelles; 3-5 buccal, two frontoterminal, 7-12 transverse and two pretransverse ventral cirri; midventral complex comprised of 10-20 pairs and two rows extending to transverse cirri; posterior part of marginal rows slightly overlapping; colorless cortical granules about 1 µm across, arranged in small groups; soil habitat. Its main ontogenetic features are: (1) in the proter, the parental adoral zone of membranelles is completely renewed by new structures; (2) in the opisthe, the oral primordium originates apokinetally, some old midventral cirri join the formation of frontoventral-transverse cirral anlagen; (3) the anlagen for marginal rows and dorsal kineties develop intrakinetally; and (4) the numerous macronuclear nodules fuse into a single mass before dividing. Phylogenetic analyses based on the SSU rDNA sequence suggest the non-monophyly of the genus Bakuella.

New contributions to the taxonomy of urostylid ciliates (Ciliophora, Hypotrichia), with establishment of a new genus and new species.

Hypotrichia, one of the most complex and highly differentiated groups in Ciliophora, has been the object of extensive studies, especially in recent years. Nevertheless, methodological difficulties and insufficient faunistic studies have limited our understanding of their biodiversity and phylogeny. In this study, one novel urostylid ciliate, Pseudoholosticha zhaoi nov. gen., nov. spec. and two populations of Anteholosticha monilata (Kahl, 1928) Berger, 2003, type species of the latter genus, are studied using an integrative approach (live observation, protargol impregnation, scanning electron microscopy, and phylogenetic analysis) to provide further insights into the diversity, classification, and phylogeny of this group of ciliates. Pseudoholosticha nov. gen. can be separated from other morphologically similar genera mainly by the absence of buccal and caudal cirri. A key to 12 morphologically similar genera and illustrations of their cirral patterns are provided. The validation of the new genus and new species is supported by both morphological and phylogenetic analyses. The first 18S rRNA gene sequence of A. monilata, with detailed morphological data, provided a reliable clarification of A. monilata populations and corroborated the phylogenetic position of the type species of the polyphyletic genus Anteholosticha.

Molecular phylogeny of a new gonostomatid ciliate revealing a discrepancy between interphasic and cell divisional patterns (Ciliophora, Hypotricha).

The morphology, ontogenesis, and molecular phylogeny of a new ciliate, Gonostomum koreanum n. sp., discovered in a terrestrial moss sample from South Korea, were investigated. Morphologically, it is characterized by a gonostomatid oral apparatus, two macronuclear nodules, six frontoventral rows, the two rightmost of which (frontoventral rows V and VI) extend posteriorly to near pretransverse and transverse cirri, and three dorsal kineties each with a single caudal cirrus posteriorly. The new species is easily confused with members of the genus Metagonostomum because of the long frontoterminal cirral row but differs mainly in the de novo (vs. intrakinetal) origin of anlage VI, a character found only in Gonostomum and Paragonostomum. To solve the discrepancy between the interphasic and ontogenetic patterns, we additionally performed morphological and multigene analyses on three gonostomatid species, namely Gonostomum koreanum n. sp. and its morphologically (M. gonostomoida) and ontogenetically (G. kuehnelti) most similar species. The multigene analyses show that the new species is closely related to G. kuehnelti and the core gonostomatids consists of five groups based on the origin of the frontoventral rows.

Morphology, ontogenesis and molecular phylogeny of a new saline soil ciliate, Uroleptoides salina nov. spec. (Ciliophora, Hypotrichia).

The morphology, morphogenesis and molecular phylogeny of a new saline soil hypotrich ciliate, Uroleptoides salina nov. spec., discovered from China, was investigated. The new species is characterized as follows: body 150-215 × 40-50 µm in vivo, slender and highly flexible; usually four ellipsoidal macronuclear nodules; contractile vacuole absent; cortical granules absent; endosymbiotic algae present; amphisiellid median cirral row consists of 14-25 cirri and terminates about 47% down length of body; usually three buccal cirri and 3-13 cirri left of anterior portion of amphisiellid median cirral row; 3-5 transverse cirri. Morphogenesis during binary fission is characterized by: (1) the parental adoral zone of membranelles is retained completely, parental paroral contributes to the formation of the undulating membranes anlage for the proter; (2) the oral primordium of the opisthe is formed apokinetally; and (3) the amphisiellid median cirral row is formed from two anlagen. Phylogenetic analyses based on SSU rDNA sequence data show that Uroleptoides salina nov. spec. has a close relationship with its morphologically similar species, U. longiseries, U. magnigranulosus, Orthamphisiella breviseries, and Parabistichella variabilis.

Morphology and molecular phylogeny of two new soil ciliates, Hemiurosomoida warreni nov. spec. and Hemiurosoma clampi nov. spec. (Ciliophora, Hypotrichia) from Tibet.

Two novel hypotrichous ciliates, Hemiurosomoida warreni nov. spec. and Hemiurosoma clampi nov. spec., isolated from soil in the Lhalu Wetland and Motuo Virgin Forest in Tibet, respectively, were investigated using live observation and protargol staining. Hemiurosomoida warreni nov. spec. strongly resembles the type species H. longa but can be distinguished by its body size in vivo (110-145 × 30-40 µm vs. 50-100 × 18-40 µm), number of adoral membranelles (25-38 vs. 15-22), and numbers of right (29-39 vs. 14-23) and left (26-35 vs. 13-23) marginal cirri, transverse cirri (3 vs. 4 or 5) and macronuclear nodules (4-8 vs. 2). Hemiurosoma clampi nov. spec. is characterized by its vermiform body shape, colourless cortical granules distributed in irregular rows, two macronuclear nodules, three frontal cirri, one buccal cirrus, four frontoventral cirri ranged in a line, two transverse cirri, lacking postoral ventral and pretransverse ventral cirri, and marginal rows that are not posteriorly confluent. Phylogenetic analyses based on SSU rDNA gene sequences suggest that Hemiurosomoida is not monophyletic. A close relationship is revealed between Hemiurosomoida warreni nov. spec., Parakahilella macrostoma, Hemiurosoma clampi nov. spec., and the type species Hemiurosoma terricola. As expected, all these species are classified within the "non-oxytrichid Dorsomarginalia".

Morphology and ontogenesis of two new Hemiholosticha species (Ciliophora, Hypotrichia, Hemiholostichidae nov. fam.).

The morphology and ontogenesis of two new hypotrich ciliates, Hemiholosticha solitaria and Hemiholosticha germanica, were studied using live observation, protargol impregnation, and scanning electron microscopy. Both species share a medium-sized, almost globular body with a short anterior projection; two macronuclear nodules with a single micronucleus in between; a central contractile vacuole; three or four ventral, one postoral, one right and one left marginal cirral row; and three dorsal kineties extending along ribs. However, H. germanica is distinguished from congeners by a higher number of cirri in ventral rows R1 and R2 (3-6 vs. 2 cirri in each row). Hemiholosticha solitaria differs from congeners by having four (vs. three) ventral cirral rows and by the lack (vs. presence) of intracellular green algae. The ontogenesis of H. solitaria follows the H. pantanalensis mode in that (i) the oral primordium develops in a deep pouch and generates the first two cirral streaks in addition to adoral membranelles and undulating membranes, (ii) the undulating membrane anlage does not produce any cirri, and (iii) the longitudinal ventral cirral row R3 originates from two anlagen. The ontogenetic peculiarities along with the 18S rRNA gene phylogenies suggest classification of Hemiholosticha, Psilotrichides, and Urospinula into a new family, Hemiholostichidae.

Morphology, morphogenesis and molecular phylogeny of a novel soil ciliate, Afrokahliella paramacrostoma n. sp. (Ciliophora, Hypotrichia).

A soil hypotrich ciliate, Afrokahliella paramacrostoma n. sp., was discovered in China. Its morphology, morphogenesis and molecular phylogeny were investigated using standard methods. The new species is characterized as follows: body about 140-180 × 60-70 µm in vivo, cortical granules absent, contractile vacuole positioned about 40% down length of body, 5-9 macronuclear nodules, 34-49 adoral membranelles, 3-5 buccal and 3-6 parabuccal cirri, usually two frontoventral rows, three or four left and two or three right marginal rows, three dorsal kineties and one dorsomarginal kinety; 1-3 and one or two caudal cirri located at the ends of dorsal kineties 1 and 2, respectively. The ontogenetic process is characterized by: (1) the marginal anlagen on each side develop in the outer right and the inner left marginal rows, respectively; (2) five frontoventral-transverse cirral anlagen, anlagen II-IV develop in secondary mode; (3) dorsal morphogenesis follows a typical Urosomoida-pattern, no parental dorsal kineties are retained; (4) caudal cirri are generated at the ends of dorsal kineties 1 and 2. Phylogenetic analyses based on SSU rDNA sequence data reveals that Afrokahliella paramacrostoma n. sp. is closely related to Parakahliella macrostoma and Hemiurosomoida longa.

Morphology and taxonomy of Pseudokeronopsis rubra (Ehrenberg, 1836) and Pseudokeronopsis parasongi sp. nov. (Ciliophora, Hypotrichia, Urostylida) from the Yellow Sea.

Pseudokeronopsis rubra (Ehrenberg, 1836), type species of Pseudokeronopsis, is a widely reported but taxonomically confused species with most available sequences either misidentified or dubious. Based on the Yellow Sea population, of which the molecular data have been analyzed in a previous work, we redescribe P. rubra and try to clarify the identifications of the populations designated as P. rubra or those allocated to this species. Nonetheless, there is still a need of integrated investigation of the species combing both morphological and molecular data based on specimens from its type locality. Recent molecular analyses indicate a Yellow Sea population of Pseudokeronopsis (marked as P. cf. songi) likely represents a new species. Here we describe it as P. parasongi sp. nov. based on the specimens from the same population. The new species possesses a yellow-brownish body with both pigmented cortical granules and colorless blood cell-shaped structures underneath the body surface, invariably one contractile vacuole at 66% of body length and occasionally a second one at 25%, and the midventral complex terminating 2-23 µm ahead of 2-4 transverse cirri. It differs from closely related species by its yellow-brownish body color and separates from them in the ITS1-5.8S-ITS2 and COI genetic distances.

The insights into the systematic relationship of Gastrostyla-affinitive genera, with report on a new saline soil ciliate genus and new species (Protozoa, Ciliophora).

BACKGROUND: Hypotrichia are a group with the most complex morphology and morphogenesis within the ciliated protists. The classification of Gastrostyla-like species, a taxonomically difficult group of hypotrichs with a common ventral cirral pattern but various dorsal and ontogenetic patterns, is poorly understood. Hence, systematic relationships within this group and with other taxa in the subclass Hypotrichia remain unresolved. RESULTS: 18S rRNA gene sequence of a new Gastrostyla-like taxon was obtained. Phylogenetic analyses based on the 18S rRNA gene sequences indicate that this ciliate represents a new genus that is closely related to Heterourosomoida and Kleinstyla within the oxytrichid clade of the Hypotrichia. However, the position of this cluster remains unresolved. All three genera deviate from the typical oxytrichids by their incomplete (or lack of) dorsal kinety fragmentation during morphogenesis. Morphology and morphogenesis of this newly discovered form, Heterogastrostyla salina nov. gen., nov. spec., are described. Heterogastrostyla nov. gen., is characterised as follows: more than 18 fronto-ventral-transverse cirri, cirral anlagen V and VI develop pretransverse cirri, and dorsal ciliature in Urosomoida-like pattern. CONCLUSIONS: Similar to the CEUU-hypothesis about convergent evolution of urostylids and uroleptids, we speculate that the shared ventral cirral patterns of Gastrostyla-like taxa might have resulted from convergent evolution.

An Integrative Investigation of Parabistichella variabilis (Protista, Ciliophora, Hypotrichia) Including Its General Morphology, Ultrastructure, Ontogenesis, and Molecular Phylogeny.

Hypotrichs are a highly differentiated and very diverse group of ciliated protists. Their systematics and taxonomy are challenging and call for detailed investigations on their general morphology, ultrastructure, ontogenesis, and molecular phylogeny. Here, a comprehensive study is conducted on a brackish water population of Parabistichella variabilis using light and electron microscopy and phylogenetic analyses based on small subunit ribosomal DNA sequence data. Its morphology, including the infraciliature, pellicle, nuclei, buccal seal, and extrusomes, is documented. The present findings indicate that in P. variabilis: (i) the cortical granules are extrusomes, which differ from those of other hypotrichs; (ii) the buccal seal is bounded by the plasma membrane and contains a single layer of longitudinal microtubules; (iii) two contractile vacuoles might be present rather than one; and (iv) the pharyngeal disks are bounded by a single membrane. Early-to-middle stages of ontogenesis are described for the first time, enabling the complete characterization of this process. Phylogenetic analyses indicate that Parabistichella variabilis is closely related to several species from different genera, such as Orthoamphisiella breviseries, Uroleptoides magnigranulosus, and Tachysoma pellionellum. However, ultrastructural and gene sequence data for more taxa are needed in order to resolve the systematics of Parabistichella.

Reevaluation of the 'well-known' Paraurostyla weissei complex, with notes on the ontogenesis of a new Paraurostyla species (Ciliophora, Hypotrichia).

Two hypotrichous ciliates, Paraurostyla wuhanensis nov. spec. from Wuhan (China) and a new North American population of the Paraurostyla weissei complex, were studied based on live observations and protargol impregnation. Paraurostyla wuhanensis nov. spec. differs from the congeners by the combined features of having six or seven frontal cirri, 2-4 frontoventral cirri, 5-7 ventral rows, and yellow-greenish cortical granules. Ontogenesis proceeds as in the type species, except that fewer frontoventral cirri are formed in the new species. The morphology of the new population of the Paraurostyla weissei complex corresponds well with other American populations. In the phylogenetic trees based on the 18S rRNA gene, Paraurostyla sequences nest in a large clade together with Apoamphisiella and Notohymena. Monophyly of Paraurostyla is rejected by the results of the approximately unbiased test analyses. Morphological, morphogenetic, and phylogenetic analyses show a close relationship between the North American populations of the Paraurostyla weissei complex and Apoamphisiella, indicating that the taxonomic position of the former needs to be reassigned. A new combination, viz. Apoamphisiella polymicronucleata (Merriman, 1937) comb. nov., a reevaluation of the Paraurostyla weissei complex, and emended diagnoses of Paraurostyla and Apoamphisiella, are provided.