Morphological and molecular characterization of two Trichodina (Ciliophora,Peritrichia) species from freshwater fishes in China

In the present study, we provide morphological and molecular characterization of two Trichodina species, T. acuta Lom, 1970 and T. funduli Wellborn, 1967, isolated from koi (Cyprinus carpio) and loach (Paramisgurnus dabryanus), respectively. Morphological characters of the two Trichodina species were mainly investigated on the basis of dry silver nitrate-impregnated specimens. Both species are medium-sized and possess well-developed denticles comprising strongly sickle-shaped blades, well-developed central parts, and straight rays. Trichodina acuta can be easily distinguished from the other Trichodina species that possess a clear central circle by the well-developed sharp blade apophysis, and the gap between ray tip and central circle. Trichodina funduli is a poorly known species that is easily confused with T. heterodentata Duncan, 1977, however the latter species has thinner denticles. The small subunit ribosomal RNA gene sequences of Trichodina acuta and T. funduli were incorporated into phylogenetic analyses. Our findings suggest that the phylogenetic lineage of trichodinids might not correspond with their living environments, host species or even some morphological characteristics.     

Morphological and Molecular Identification of the New Species,Trichodina pseudoheterodentata sp. n. (Ciliophora,Mobilida, Trichodinidae) from the Channel Catfish,Ictalurus punctatus,in Chongqing China

It is difficult to differentiate similar trichodinids solely based on morphological examination, thus other identification methods, such as molecular identification, are necessary for identification. One mobilid ciliate named Trichodina pseudoheterodentata sp. n. was isolated from the gills of channel catfish, Ictalurus punctatus, in Chongqing, China. In the present study, its SSU rDNA was sequenced for the first time. Based on the results from both morphological identification and SSU rDNA sequencing, the new species was identified and compared with similar species. The morphological analysis revealed that T. pseudoheterodentata is a large Trichodina species (cell diameter 73.0–82.5 μm) and possesses robust denticles with broad blades and well‐developed blade connections. Characterization of its primary and secondary SSU rDNA structures indicated that T. pseudoheterodentata was distinctly different from congeneric species in H12, H15, E10_1, and V4 regions. Phylogenetic analysis revealed that the genetic distances among the new species and similar species reached interspecific levels, furthermore, the phylogenetic study also validated the identification of T. pseudoheterodentata and its placement in the genus Trichodina.     

Phylogenetic Analyses of Trichodinids (Ciliophora, Oligohymenophora) Inferred from 18S rRNA Gene Sequence Data

Partial 18S rRNA gene sequences of the three trichodinids, namely Trichodina modesta Lom, 1970, Trichodina paraheterodentata Tang and Zhao 2012. and Trichodinella epizootica (Raabe 1950) ?rámek-Hu?ek, 1953, were acquired and used to construct phylogenetic trees. The results revealed that Trichodinella epizootica clustered with Trichodinella sp.; Trichodina paraheterodentata Tang and Zhao 2012 was sister to the clade composed of Trichodina heterodentata Duncan, 1977 and Trichodina nobilis Chen, 1963; Trichodina modesta Lom, 1970 clustered with Trichodina reticulata Hirschman and Partsch, 1955. The branching order of species within the Mobilia clade was closely correlated with GC content. Furthermore, blade morphology was also found to be the primary morphological character in determining the phylogenetic relationships among members of the genus Trichodina. The present findings suggest that the genus Trichodina is paraphyletic when species of Trichodinella are included in the analyses.     

A new species of <Emphasis Type="Italic">Trichodina</Emphasis> Ehrenberg, 1830 (Ciliophora: Trichodinidae) infecting farmed <Emphasis Type="Italic">Clarias gariepinus</Emphasis> (Burchell) (Siluriformes: Clariidae) in Cuba

A new species of Trichodina Ehrenberg, 1830 collected from the skin and fins of farmed North African catfish Clarias gariepinus (Burchell) fingerlings, is described. The new species can be distinguished from other trichodinids by the characteristics of the adhesive disc, especially by the great number of denticles. Trichodina merciae n. sp. is morphologically similar to T. renicola (Mueller, 1931) and T. marplatensis Martorelli, Marcotegui & Alda, 2008, in the number of denticles, but differs in the morphometric data, denticle morphology, environment and location. Trichodina merciae n. sp. has broad sickle-shaped blades and thin, straight rays, while T. marplatensis has broad club-shaped blades and wide S-shaped rays. Besides, denticle length, blade length, ray length, width of central part and denticle span of the new species are greater than T. marplatensis. However, the diameter of denticle ring and the diameter of the central area in T. marplatensis is larger than the ones in T. merciae n. sp. This is the first record of freshwater ectoparasite trichodinid with an average number of denticles greater than 50.     

Trichodinid ectoparasites (Ciliophora: Peritrichida) of wild and cultured freshwater fishes in Taiwan,with notes on their origin

During 1988 parasitological surveys were carried out on the freshwater fish fauna on the island of Taiwan in 12 localities that included fishery stations, reservoirs, fish markets as well as natural rivers and swamps. A total of 28 fish species were examined, representing 14 families. This paper reports on the parasites of the genus Trichodina (Ciliophora, Peritrichida) of Taiwan. Fourteen trichodinid species were recorded of which five are new. Trichodinids collected in this study can be divided into four categories, based on their host association and possible region of origin. The first category includes trichodinids with an African origin, T. centrostrigeata Basson, Van As & Paperna, 1983, T. heterodentata Duncan, 1977, T. compacta Van As & Basson, 1989 and possibly T. canton n. sp. The second category is species with an Eurasian origin and includes T. acuta Lom, 1961, T. modesta Lom, 1970, T. mutabilis Hirschmann & Partsch, 1955, T. nigra Lom, 1961 and T. nobilis Chan, 1963. The third category comprises one species, T. maritinkae Basson & Van As, 1990, with an Afro-Asian distribution. The last category is composed of new species that are most likely native to Asia: these are T. matsu n. sp., T. mandarin n. sp., T. wulai n. sp. and T. pagoda n. sp.     

Trichodinid Protozoa (Ciliophora: Peritrichida) from Freshwater Fishes of Rybinsk Reservoir,USSR1

During the spring of 1981, trichodinid protozoa were collected from the gills or urinary bladder of the following species of fishes of Rybinsk Reservoir, USSR, located on the upper reaches of the Volga River system: Esox lucius, Rutilus rutilus, Leuciscus idus, Blicca bjoerkna, Abramis ballerus, Pelecus cultratus, Lota lota, and Perca fluviatilis. A total of 13 species of Trichodinidae was recovered from Rybinsk fishes, among which are three new species of the genus Trichodina: T. izyumovae n. sp. from the gills of L. idus, T. borokensis n. sp. from the gills of P. cultratus, and T. kupermani n. sp. from the gills of A. ballerus. Other species of trichodinids reported are T. modesta Lom, 1980; T. nigra Lom, 1969; T. pediculus (O. F. Müller, 1786); T. prowazeki Grupcheva & Lom, 1980; T. rostrata Kulemina, 1968; T. urinaria Dogiel, 1940; T. ophiocephalus Kostenko & Karaev, 1976; Paratrichodina incisa (Lom, 1959); Trichodinella epizootica (Raabe, 1950); and Tripartiella copiosa Lom, 1959. Photomicrographs and morphometric data are presented for each species and aspects of their host and geographic distribution discussed. Trichodina algonquinensis Li & Desser, 1983 is considered a synonym of T. urinaria Dogiel, 1940.     

An SEM study of adhesive disc skeletal structures isolated from trichodinids (Ciliophora: Peritrichida) of the genera Trichodina Ehrenberg, 1838 and Paratrichodina Lom, 1963

Specimens of Trichodina domerguei Wallengren, 1897, T. intermedia (Lom, 1961) and Paratrichodina incissa (Lom, 1959) were sonicated to liberate skeletal components of the adhesive disc. This enabled SEM observation of the taxonomically important structures obscured in preparations of complete cells. A previously undescribed peg-like structure on the centrifugal surface of the central part of the denticles is revealed in T. domerguei. In P. incissa the ray apophysis and its supporting apophysis appear to be absent, providing an additional characteristic to discriminate it from species of the genus Trichodina Ehrenberg, 1838. From silver stained and SEM preparations of T. intermedia and P. incissa important differences in denticle blade form are apparent, underlining the value of observation of isolated skeletal structures by electron microscopy.     

Evaluating molecular support for and against the monophyly of the Peritrichia and phylogenetic relationships within the Mobilida (Ciliophora,Oligohymenophorea)

Zhan, Z., Xu, K. & Dunthorn, M. (2012). Evaluating molecular support for and against the monophyly of the Peritrichia and phylogenetic relationships within the Mobilida (Ciliophora, Oligohymenophorea). —Zoologica Scripta,00, 000–000. An ongoing debate in ciliate molecular phylogenetic analyses is whether the Peritrichia – composed of the Sessilida and the Mobilida – are monophyletic. To further investigate this group, here we increase taxon sampling with four further mobilids: Trichodinella sp., Trichodina pectenis, Urceolaria serpularum and Urceolaria korschelti. Results show that support for and against monophyly of the peritrichs depends on different methods of alignment, and different methods of masking ambiguously aligned nucleotide positions. Results from constrained analyses also are dependent on different alignment and masking methods. We propose that there is no well‐supported SSU‐rDNA evidence supporting for and against the removal of the mobilids from the peritrichs. Within the Mobilida, we find that Trichodinella nest within Trichodina, Urceolaria branch basally, and Trichodina are non‐monophyletic, and suggest that Leiotrocha be formally transferred into Urceolaria. In addition, the data support that Urceolaria‐like denticles and Urceolaria‐/Trichodina‐like adoral ciliary spiral are plesiomorphic conditions within the mobilids.     

寄生于重庆地区鲢、鳙及草鱼五种外寄生车轮虫的描述

对采自重庆地区寄生于鲢、鳙及草鱼鳃上的5种外寄生车轮虫进行了形态学及分类学研究,其分别隶属于：车轮虫属、小车轮虫属及三分虫属,其中包含1新种和中国2新纪录。即钝圆车轮虫Trichodina obtusiformis sp．nov．,急尖车轮虫T．acuta Lom,1961,显著车轮虫T．nobilis Chen,1963,周丛小车轮虫Trichodinella epizootica（Rabbe,1950）Sramek-Husek,1953和大型三分虫Tripartiella macrosoma Basson ＆ Van As,1987。文中对新种进行了详细的描述并提供了新种及已知种的附着盘银染标本之显微照片及齿体线条图。     

First Record of the Peritrich Trichodina diaptomiBasson and Van As, 1991 (Protozoa: Ciliophora) on a South American Calanoid Notodiaptomus deitersi (Poppe, 1890) (Crustacea: Copepoda)

ABSTRACT. The ciliate Trichodina was recorded on the calanoid Notodiaptomus deitersi in a shallow, eutrophic reservoir, located in the centre‐west of Brazil. The species was confirmed as Trichodina diaptomi, a species widely distributed in Europe, Africa, Asia and Australia. It was observed moving freely over the carapace of the copepod, and using its adhesive disc. This is the first record for the species in the Americas.     

Grellamoeba robusta gen. n., sp. n., a possible member of the family Acramoebidae Smirnov,Nassonova et Cavalier-Smith, 2008

A strain of naked amoeba isolated from pikeperch (Sander lucioperca (L.)) kidney tissue has been characterized using light- and transmission electron microscopy. Sequencing of SSU rDNA and phylogenetic analysis based on a broad dataset of sequences completed our study. All data obtained suggest that this strain belongs to a species that has not been described before. As none of the existing genera of amoebae is applicable to this organism, the new genus Grellamoeba is established and the type species Grellamoeba robusta is described. Although the phylogenetic position of the SSU rDNA sequence of the type strain of G. robusta is sensitive to the method of analysis applied, a tendency to group with Acramoeba dendroida Smirnov, Nassonova et Cavalier-Smith, 2008 is evident.     

南中国海野生海水鱼类外寄生车轮虫新记录

研究对采自南中国海的7种海水鱼类进行了调查研究, 并分离获得4种隶属于车轮虫属的种类,分别为浦氏车轮虫Trichodina puytoraci Lom, 1962; 日本车轮虫Trichodina japonica Imai, et al., 1991; 直钩车轮虫Trichodina rectuncinata Raabe, 1958 与鲀车轮虫Trichodina fugu Imai, et al., 1997。研究发现鲀车轮虫Trichodina fugu Imai, et al., 1997为寄主鱼虫纹东方鲀Takifugu vermicularis的病原, 且能在野生条件下将其致死。上述4种车轮虫的形态分类学数据均基于干银法标本获得。研究是对我国南中国海车轮虫的首次报道。       

大亚湾养殖鱼类寄生车轮虫的种类组成与季节动态

2017年8月至2018年7月对大亚湾8种养殖鱼类的车轮虫感染情况进行了调查, 采用Lom倡导的“统一特定方法”及Van As & Basson提出的“齿体定位描述法”对采获的车轮虫标本进行了形态鉴定与描述。结果在卵形鲳鲹(Trachinotus ovatus)、星点笛鲷(Lutjanus stellatus)、鞍带石斑鱼(Epinephelus lanceolatus)、眼斑拟石首鱼(Sciaenops ocellatus)和虎龙杂交斑[E. fuscoguttatus (♀) × E. lanceolatus (♂)]等5种鱼上共检获5种车轮虫: 亚卓车轮虫(Trichodina jadranica)、莱普斯车轮虫(T. lepsii)、长须鳠车轮虫(T. guliae)、劳牧小车轮虫(Trichodinella lomi)和斜拟车轮虫(Paratrichodina oblique); 其中卵形鲳鲹5种虫均有寄生, 虎龙杂交斑感染了4种虫, 星点笛鲷、眼斑拟石首鱼和鞍带石斑鱼分别感染了3种、2种和1种虫。周年感染数据表明: 5种虫的感染率均以卵形鲳鲹最高, 眼斑拟石首鱼上2种虫的感染率最低; 莱普斯车轮虫平均感染强度最高的宿主是虎龙杂交斑, 其他4种虫的均是卵形鲳鲹。寄生车轮虫的季节动态表明: 冬季的感染率和平均感染强度较低; 不同虫种感染高峰出现的季节不一, 其中斜拟车轮虫的感染高峰出现在秋季, 其春夏秋季的平均感染强度均大于其他4种虫。研究填补了大亚湾海区鱼类寄生车轮虫研究的空缺, 为该海区车轮虫病的诊断与防控提供了参考。     

Zschokkella hildae Auerbach, 1910: Phylogenetic position,morphology, and location in cultured Atlantic cod

The myxozoan Zschokkella hildae Auerbach, 1910, was detected with a prevalence of 100% in cultured Atlantic cod, Gadus morhua L. aged 1+ from a culture facility on the west coast of Scotland. Sporogonic stages of Z. hildae, plasmodia producing 2–5 mature spores, were located predominantly in the collecting ducts and ureters of the kidney, and spores were present in the urine collected from the bladder. Less frequently, plasmodia were detected in the interstitial tissue of the kidney. The parasite prevalence in cultured fish was considerably higher than reported in wild fish but no obvious signs of pathology were detected. SSU rDNA sequencing and phylogenetic analysis showed that Z. hildae is closely related to a Sinuolinea sp. from the urinary system of turbot, Psetta maxima (L.), and that these two species, together with other myxozoans from the urinary system of marine fish cluster together in a sub-clade of the recognised marine clade of myxozoans. This sub-clade is characterised by a specific linear expansion segment, helix E23_15 in the secondary structure of variable region V4 of the SSU rDNA. Z. hildae and Sinuolinea sp. show extraordinary large linear expansion segment in both V4 and V7 and an important number of complementary base changes in the conservative regions of the SSU rDNA, indicating considerable evolutionary changes in the SSU rDNA of these species when compared with other myxozoans from the marine environment.     

New taxa refresh the phylogeny and classification of pleurostomatid ciliates (Ciliophora,Litostomatea)

A high diversity of pleurostomatid ciliates has been discovered in the last decade, and their systematics needs to be improved in the light of new findings concerning their morphology and molecular phylogeny. In this work, a new genus, Protolitonotus gen. n., and two new species, Protolitonotus magnus sp. n. and Protolitonotus longus sp. n., were studied. Furthermore, 19 novel nucleotide sequences of SSU rDNA, LSU rDNA and ITS1‐5.8S‐ITS2 were collected to determine the phylogenetic relationships and systematic positions of the pleurostomatid ciliates in this study. Based on both molecular and morphological data, the results demonstrated that: (i) as disclosed by the sequence analysis of SSU rDNA, LSU rDNA and ITS1‐5.8S‐ITS2, Protolitonotus gen. n. is sister to all other pleurostomatids and thus represents an independent lineage and a separate family, Protolitonotidae fam. n., which is defined by the presence of a semi‐suture formed by the right somatic kineties near the dorsal margin of the body; (ii) the families Litonotidae and Kentrophyllidae are both monophyletic based on both SSU rDNA and LSU rDNA sequences, whereas Amphileptidae are non‐monophyletic in trees inferred from SSU rDNA sequences; and (iii) the genera Loxophyllum and Kentrophyllum are both monophyletic, whereas Litonotus is non‐monophyletic based on SSU rDNA analyses. ITS1‐5.8S‐ITS2 sequence data were used for the phylogenetic analyses of pleurostomatids for the first time; however, species relationships were less well resolved than in the SSU rDNA and LSU rDNA trees. In addition, a major revision to the classification of the order Pleurostomatida is suggested and a key to its families and genera is provided.     

Morphological,histological and molecular characterization of three Myxobolus species (Cnidaria: Myxosporea) from silver carp Hypophthalmichthys molitrix Valenciennes and bighead carp Hypophthalmichthys nobilis Richardson in China

Three Myxobolus species were obtained from silver carp Hypophthalmichthys molitrix Valenciennes and bighead carp Hypophthalmichthys nobilis Richardson in China. In the present study, we supplemented their taxonomic characteristics by the morphological, histological and molecular methods. Myxobolus kiuchowensis Chen in Chen et Ma, 1998 formed small ellipsoidal plasmodia in the intestinal wall of bighead carp. Its spores appeared asymmetrical obovate in frontal view and fusiform in lateral view. Tiny mamillary protrusion in the anterior of some spores was observed. Two pyriform polar capsules were unequal. Histologically, M. kiuchowensis infected the tunica muscularis of host intestine. Myxobolus abitus Li et Nie, 1973 formed sausage–like plasmodia in the gills of silver carp. Its spores appeared oblate in frontal view and fusiform in lateral view. Two pyriform polar capsules were unequal and an obvious inter–capsule appendix was observed. Histological examination revealed that M. abitus developed in the interlamellar–epithelium of host gills. Myxobolus pavlovskii (Akhmerov, 1954) Landsberg et Lom, 1991 formed sausage–like plasmodia both in the gills of silver carp and bighead carp. Spores of M. pavlovskii were proximate oval in frontal view and fusiform in lateral view. Two pyriform polar capsules were unequal. The BLAST search indicated the SSU rDNA sequences of M. kiuchowensis and M. abitus were not identical to any sequence, however, the SSU rDNA sequences of M. pavlovskii were identical to that of M. pavlovskii recorded previously. Phylogenetic analysis showed that the present three species robustly clustered together in Cyprinid group and Asia group.