Morphology and Phylogenetic Placement of Three New Zoothamnium species (Ciliophora: Peritrichia) from Coastal Waters of Southern China

The morphology, infraciliature, and silverline system of three peritrichous ciliates, Zoothamnium bucciniiformum sp. n., Zoothamnium florens sp. n., and Zoothamnium zhanjiangense sp. n., were investigated based on both living and silver‐stained specimens. Zoothamnium bucciniiformum sp. n., collected from coastal waters (salinity 30‰) off Zhanjiang, southern China, can be distinguished by the following characters: dichotomously branched stalk, peristomial lip with medial circumferential infolding, contractile vacuole apically positioned, 32–49 silverlines between the anterior end and the aboral trochal band, 15–26 between the aboral trochal band and the scopula; two kineties in peniculus 3, not parallel to each other. Zoothamnium florens sp. n., collected from a mangrove wetland (salinity 13‰) off Zhanjiang, is characterized by its large conical zooid, tuberculate peristomial lip, asymmetrical dichotomously branched colony, 59–81 silverlines between the anterior end and the aboral trochal band and 29–36 between the aboral trochal band and the scopula. Zoothamnium zhanjiangense, collected from a mangrove wetland (salinity about 9.5‰) off Zhanjiang, differs from its congeners by the alternately branched stalk, peristomial lip with medial circumferential infolding, 40–63 silverlines from the peristomial area to the aboral trochal band and 13–24 from the aboral trochal band to the scopula. The comparison and analysis of SSU rDNA sequences also support present identifications.     

Morphology and Phylogeny of Three Pseudovorticella Species (Ciliophora: Peritrichia) from Brackish Waters of China

The biodiversity of peritrich ciliates from brackish biotopes is rarely investigated, especially members of the genus Pseudovorticella. Here, the morphology of three species of Pseudovorticella, i.e. P. cf. vestita (Stokes, 1883) Jankowski, 1976, P. spathulata sp. n., and P. qinghaiensis sp. n. isolated from brackish waters were studied. Pseudovorticella cf. vestita is characterized by inverted bell‐shaped cell; a J‐shaped macronucleus; a single contractile vacuole ventrally located; P3 three‐rowed; pellicle striated with highly developed pellicular vesicles; 18–22 transverse silverlines between peristome and aboral trochal band, and 9–13 between aboral trochal band and scopula. Pseudovorticella spathulata sp. n. differs from its congeners by the following combination of characters: elongate‐elliptical cell; a single contractile vacuole near ventral wall of infundibulum; a J‐shaped macronucleus; P3 three‐rowed; 24–34 silverlines between oral area and aboral trochal band and 6–10 between aboral trochal band and scopula. Pseudovorticella qinghaiensis sp. n. is characterized by: cell with an oval outline; a single contractile vacuole near ventral wall of infundibulum; a C‐shaped macronucleus; P3 three‐rowed; 30–35 and 9–11 transverse silverlines above and below the trochal band, respectively. The SSU rDNA sequences of five Pseudovorticella species, namely P. annulata, P. monilata, P. parakenti, P. spathulata sp. n., and P. cf. vestita, plus that of Zoothamnium hartwigi, are reported for the first time and their evolutionary relationships are investigated. Five undefined Pseudovorticella forms are considered might be conspecific with P. monilata. Two congeners are conspecific with P. spathulata sp. n. Phylogenetic analyses based on SSU rDNA sequences reveal that Pseudovorticella is not monophyletic and Z. hartwigi clusters with its congeners as expected.     

Morphology and Phylogeny of Three New Loxophyllum Species (Ciliophora,Pleurostomatida) from Mangrove Wetlands of Southern China

Three new pleurostomatid ciliates, Loxophyllum meridionale sp. n., L. salinum sp. n., and L. planum sp. n., isolated from brackish waters of mangrove wetlands in southern China, were investigated using live observation and protargol impregnation. The main characteristic feature of L. meridionale sp. n. is the presence of three contractile vacuoles (CVs) along its dorsal margin. Loxophyllum salinum sp. n. can be identified by the presence of three closely spaced CVs along the ventral margin and 12–15 right and four to six left kineties. Loxophyllum planum sp. n. is characterized by the combination of two macronuclei, a single, subterminal CV, and the absence of cortical granules. Phylogenetic trees based on the small subunit rDNA sequence data recovered Loxophyllum as a monophyletic group within the pleurostomatids. Loxophyllum meridionale sp. n., L. salinum sp. n., and L. planum sp. n. were all nested within the Loxophyllum clade. Based on its ciliary pattern and general morphological features, Loxophyllum asetosum is transferred to the genus Amphileptus and a new combination is suggested: Amphileptus asetosus (Burkovsky 1970) comb. n. [basionym L. asetosum Burkovsky 1970].     

Morphology,Ontogeny, and Molecular Phylogeny of Two Freshwater Species of Deviata (Ciliophora,Hypotrichia) from Southern China

The morphology and partial morphogenesis of two freshwater hypotrichous ciliates, Deviata brasiliensis Siqueira‐Castro et al., 2009 and Deviata rositae Küppers et al., 2007, isolated from southern China, were investigated using live observation and protargol staining. Our populations resemble the original ones in terms of their live characters and ciliary patterns. The main determinable morphogenetic features of D. brasiliensis basically correspond with those of the type population. However, the origin of anlage V for either proter or opisthe is ambiguous: whether anlage V for the proter originates from parental frontoventral row 2 (the same as in the original population) or parental frontoventral row 3 (the same as in Deviata abbrevescens) or even de novo is not clear; the anlage V for the opisthe is possibly derived from frontoventral row 3 and further migrates to frontoventral row 2, like that in D. abbrevescens. In addition, the SSU rRNA gene was first sequenced for both species. Molecular phylogenetic analyses suggest that the genus Deviata is non‐monophyletic and has a close relationship with Perisincirra paucicirrata.     

The Morphology of Three Loxophyllum Species (Ciliophora,Pleurostomatida) from Southern China,L. lembum sp. n., L. vesiculosum sp. n. and L. perihoplophorum Buddenbrock, 1920, with Notes on the Molecular Phylogeny of Loxophyllum

Two new pleurostomatid ciliates, Loxophyllum lembum sp. n., L. vesiculosum sp. n., and the poorly known L. perihoplophorum Buddenbrock, 1920, isolated from brackish waters in coastal regions of southern China, are described following observations of live cells and protargol‐impregnated specimens. Loxophyllum lembum sp. n. is distinguished by a combination of characters including two macronuclear nodules, 6–9 contractile vacuoles along the ventral margin, 11–14 right and 6–8 left kineties and the presence of cortical granules. Loxophyllum vesiculosum sp. n. differs from its congeners mainly by the unique distribution of contractile vacuoles, several of which lie along the dorsal margin and one on the ventral margin, and 15–21 right and 6–8 left kineties. Loxophyllum perihoplophorum is characterized by its large cell size (350–450 μm long in vivo), 3–5 contractile vacuoles along the dorsal margin in the posterior region of the body, and 19–23 right and 7–9 left kineties. An improved diagnosis of L. perihoplophorum is provided. The SSU rDNA sequence of L. perihoplophorum is reported for the first time and its molecular phylogeny is analyzed. Maximum likelihood and Bayesian inference analyses of SSU rDNA sequence data recover the monophyly both of the order Pleurostomatida and of the genus Loxophyllum.     

Morphology and Molecular Phylogeny of Two Poorly Known Species of Protocruzia (Ciliophora: Protocruziida)

The ciliate genus Protocruzia is a highly confused group, which was formerly placed in the class Heterotrichea or Karyorelictea, and is according to the most recent system tentatively assigned to the class Spirotrichea. In the present study, the morphology, ciliary pattern, and molecular phylogeny of two poorly known species, Protocruzia tuzeti Villeneuve‐Brachon, 1940, and Protocruzia granulosa Kahl, 1933, isolated from coastal waters of China, were investigated. Protocruzia tuzeti differs from its congeners mainly in possessing 6 adoral membranelles, 8–11 somatic kineties, and postoral dikinetids. Protocruzia granulosa is characterized by its extremely slender body, three postoral kineties, and 13 or 14 somatic kineties. The morphogenesis of P. granulosa is similar to that of P. tuzeti, especially in the parakinetal mode of stomatogenesis and the reorganization of the parental paroral membrane; however, more than one somatic kinety joins in the formation of the oral primordium in P. granulosa. Phylogenetic analyses based on small subunit ribosomal RNA gene revealed that six Protocruzia species form a fully supported clade that does not belong to any ciliate class; therefore, our data support the establishment of the class Protocruziea Gao et al. (Sci. Rep., 6, 2016, 24874).     

New Species of Lagenophrys (Ciliophora,Peritrichia) from New Zealand and Australia1

Lagenophrys novazealandae n. sp. occurs on the gills of Paranephrops zealandicus, a parastacid crayfish from New Zealand. The new species has the hemispheroidal lorica most common among members of its genus and is distinguished by its possession of large tubercles on the thickened edge of the anterior lip of the lorica aperture, a deep cleft in the left side of the lip's edge, and a ovoid to reniform macronucleus located in the right-hand part of the body. It is probable that an as yet unnamed species of Lagenophrys known to occur on another species of Paranephrops in New Zealand is distinct from L. novazealandae but phylogenetically related to it. Lagenophrys petila n. sp. occurs on setae of Parastacoides tasmanicus, a parastacid from Tasmania. The new species has an ovoid lorica tapering to a slender pseudostalk at the posterior end, a type of lorica possessed by only two other members of its genus that also attach to their host's setae. It is distinguished from the other ovoid species by the proportions of the lorica, the extreme shortness of the lips of the lorica aperture, and an ovoid macronucleus located in the right, anterior part of the body. Clefts in the lips of L. novazealandae and other members of Lagenophrys may function as points of flexure to allow the lips to bend in ways that accommodate interspecific differences in the size of the epistomial disk and its operation during suspension feeding.     

Parastrombidinopsis minima n. sp. (Ciliophora: Oligotrichia) from the Coastal Waters of Northeastern Taiwan: Morphology and Small Subunit Ribosomal DNA Sequence

ABSTRACT. Parastrombidinopsis minima n. sp. is investigated, using live observations, protargol preparations, and molecular data. In living cells, the ranges of cell length are 85–95 μm, cell width 60–70 μm, and oral diameter 40–50 μm. In protargol‐impregnated specimens, cell length ranges between 43 and 71 μm, cell width between 23 and 42 μm, and oral diameter between 13 and 24 μm. The numbers of external oral polykinetids are 12–16 and of somatic kineties are 11–13. There are always two ovoid macronuclei (9–16 × 4–9 μm). Based on the analysis of morphologic data, the new species can be placed in the family Strombidinopsidae, but based on the small subunit rRNA gene sequence data, the Parastrombidinopsis species are more closely associated with strobilidiids and tintinnids.     

Epistylis portoalegrensis n. sp. (Ciliophora,Peritrichia): A New Freshwater Ciliate Species from Southern Brazil

The peritrich ciliate Epistylis portoalegrensis n. sp. was found in two bodies of freshwater located in Porto Alegre, Southern Brazil. Morphological features were investigated using live and protargol‐stained specimens. The zooids presented a vase to cylindrical shape narrowed at the scopula, and a mean size of 131 × 37 μm in vivo. A C‐shaped macronucleus lay in the middle of the cell close to a single contractile vacuole. The oral infraciliature was typical for the genus, with all infundibular polykineties composed by three distinct rows of kinetosomes. Colonies are often nonbranched with no lateral stalk, carrying several zooids stemming from a single point. Specimens from the two sampling sites showed identical arrangement of the infraciliature, similar morphometry, identical 18S rDNA sequences, and a single nucleotide difference across the more variable ITS regions. Molecular phylogenetic analyses placed E. portoalegrensis in a well‐supported clade containing other Epistylis species, within the order Vorticellida.     

Taxonomic characterization of Vorticella fuscaPrecht, 1935 and Vorticella parapulchella n. sp., two marine peritrichs (Ciliophora, Oligohymenophorea) from China

Two marine peritrich ciliates, Vorticella fuscaPrecht (1935) and Vorticella parapulchella n. sp. were discovered in the littoral zone of Qingdao, northern China. Their morphology, infraciliature, and silverline system were described using live observation and silver impregnation. The poorly known species V. fusca is redescribed, adding information about the oral infraciliature and pellicular morphology. Vorticella parapulchella n. sp. is superficially similar to Vorticella pulchellaSommer (1951) but is distinguished from it by being markedly smaller and having much more widely spaced pellicular ridges. The infundibular infraciliature of V. parapulchella is extremely unusual in having infundibular polykinety 3 reduced to two rows, one of which has almost disappeared.     

A Contribution to the Morphology and Phylogeny of Chlamydodon,with Three New Species from China (Ciliophora,Cyrtophoria)

Three new cyrtophorian ciliates isolated from coastal areas of China were described based on morphological and genetic data. The Chlamydodon mnemosyne‐like species Chlamydodon similis sp. n. differs from its congeners mainly by its number of somatic kineties. Chlamydodon oligochaetus sp. n. is distinguished from its congeners mainly by having fewer somatic kineties, and/or an elongated body shape. Chlamydodon crassidens sp. n. is characterized mainly by an inverted triangular body shape, a posteriorly interrupted cross‐striated band (5–6 μm wide), and a large cytostome. Moreover, we provided small‐subunit (SSU) rDNA sequences of C. similis sp. n. and C. oligochaetus sp. n. Maximum likelihood (ML) and Bayesian inference (BI) consistently placed C. similis sp. n. as a sister to C. paramnemosyne, but showed different branching position of C. oligochaetus sp. n., which may be due to a low taxon sampling in the Chlamydodontidae and/or an insufficient resolution of the marker gene at species level.     

A New Species of Leptolalax （Anura： Megophryidae） from Southern China

A new species, Leptolalax laui sp. nov. is described based on specimens collected from Hong Kong and Shenzhen City, Guangdong Province, China. The new species can be distinguished from other known congeners by morphological and molecular data. The new species is characterized by the following characters： 1） small size （adult males SVL 24.8.1 mm-26.7 mm）; 2） near immaculate creamy white chest and belly; 3） broad lateral fringes on toes; 4） head longer or as long as wide; 5） distinct dark brown spots in flank; 6） moderate dermal fringes on fingers; 7） brown or reddish-brown dorsum with fine round scattered tubercles; 8） thin traverse brownish-grey bars on the dorsal surface of tibia and lower arms; 9） longitudinal ridges under toes not interrupted at the articulations.     

Three New Loxophyllum Species (Ciliophora: Pleurostomatida) from China with a Brief Review of the Marine and Brackish Loxophyllum Species

Recent studies indicate that there is a high diversity of pleurostomatid ciliates in the coastal waters of China. Here, three new congeners of Loxophyllum, L. caudatum sp. n., L. rugosum sp. n., and L. chinense sp. n., are described following observations of live cells and protargol‐impregnated specimens. All three species usually have two macronuclear nodules and prominent warts along the dorsal margin formed by clustered extrusomes. In addition, L. caudatum sp. n. is characterized by its long conspicuous tail, dot‐like cortical granules, 4 or 5 left and 9 or 10 right kineties, and a single subterminal contractile vacuole. Loxophyllum rugosum sp. n. is distinguished by possessing three prominent ridges on the left side, 7–11 right and 5–7 left kineties. Loxophyllum chinense sp. n. is characterized by having several contractile vacuoles distributed along the ventral margin, 13–18 right and 6–8 left kineties. The small subunit ribosomal DNA (SSU rDNA) sequence similarities among six congeners range from 96.46% to 99.94%. Phylogenetic trees based on the SSU rDNA sequences indicate that all Loxophyllum spp. form a well‐supported monophyletic group. A brief review of the marine and brackish Loxophyllum species is supplied and one new combination, Litonotus multiplicatus (Kahl 1931) comb. n. (basionym Loxophyllum multiplicatum Kahl 1931), and one new name, Litonotus dragescoi nom. n. (basionym L. fasciolatus Dragesco 1966), are suggested.     

Choanoflagellates (Sarcomastigophora, Protozoa) from the Coastal Waters of Taiwan and Japan. I. Three New Species

Three new acanthoecidaean species collected from the coastal waters of Taiwan and Japan are described: Acanthocorbis camarensis n. sp. resembles Acanthocorbis unguiculata in lorica morphology, but differs in having regularly arranged longitudinal and transverse costae at the anterior lorica chamber, and in lacking a nail at the apical end of anterior spine; Diaphanoeca spiralifurca n. sp. is characterized by the spiral arrangement of the costal strips in the posterior half of the lorica chamber and is closely related to Diaphanoeca grandis; Stephanoeca supracostata n. sp. is closely related to Stephanoeca elegans, but differs in having an additional transverse costa at the anterior lorica chamber.     

Four New Species of the Genus Hemicriconemoides (Nematoda:Criconematidae)

Four new species of the genus Hemicriconemoides (H. californianus n.sp., H. taiwanensis n.sp., H. annulatus n. sp., and H. nitida n.sp.) are described. The range of total length of H. mangiferae is increased on the basis of specimens collected in Israel, Observations on H. mangiferae and H. litchi support the validity of H. litchi as distinct from H. mangiferae.     

Morphology and morphogenesis of Epistylis plicatilis ehrenberg, 1831 (Ciliophora,Peritrichia) from Wuhan,China

A limnetic peritrichous ciliate, Epistylis plicatilis Ehrenberg, 1831, was collected from a freshwater ditch beside Moshan Hill, Wuhan, China. Its morphology, infraciliature, and morphogenesis were investigated based on specimens examined in vivo, following staining with protargol and by scanning electron microscopy. The characteristics of the Wuhan population of E. plicatilis are as follows: 1) colonial, each colony typically comprising 30–50 individuals, with a dichotomously branched, noncontractile stalk; 2) fully expanded zooids measure 90–155 × 30–50 µm in vivo; 3) a series of 6 or 7 conspicuous folds appear in the posterior region of the zooid when it contracts; 4) single horseshoe‐shaped macronucleus oriented transversely; 5) single contractile vacuole located in peristomial region on dorsal wall of infundibulum; 6) myoneme system comprises 20–24 longitudinal fibers, peristomial disk fibers as a wreath‐like net and peristomial ring fibers; 7) narrowly spaced transverse striations on the surface of the body; 8) infundibular polykineties 1 and 2 are three‐rowed, infundibular polykinety 3 is two‐rowed; and 9) stomatogenesis is of the buccokinetal type; in the new oral apparatus, infundibular polykineties 2 and 3, the haplokinety, and the germinal kinety all originate from the germinal kinety of the parental oral apparatus whereas the polykinety and infundibular polykinety 1 originate from the parental haplokinety. An improved diagnosis of E. plicatilis is supplied. J. Morphol. 275:882–893, 2014. © 2014 Wiley Periodicals, Inc.     

Phylogeny and Genetic/Morphological Variation of Strombidinopsis minima‐like Species (Ciliophora: Choreotrichia)

Six isolates of mineral‐enveloped Strombidinopsis minima‐like species were collected from the coastal waters across several regions in Korea. Morphological observations and molecular analyses were performed. The ribosomal DNA sequences (including small subunit ribosomal DNA, internal transcriber spacer 1‐5.8S ribosomal DNA‐internal transcriber spacer 2; and part of large subunit ribosomal DNA) of these six isolates were compared. Their morphological characteristics were also compared with those of S. minima populations reported. The marked genetic differences (with a similarity range of 96.85–98.48%) in SSU rDNA among these S. minima‐like entities suggest the existence of multiple species. This finding is also supported by morphological variations detected in this study and reported in the literature (e.g. 15–32 collar membranelles in different populations). In addition, S. minima‐like species are clustered with S. batos and S. sinicum, and therefore, our SSU rDNA results support previous results suggesting that the genus Strombidinopsis is not monophyletic in origin. Further collection of morphological and molecular data may facilitate the determination of a new genus carrying mineral‐enveloped Strombidinopsis species.