Small Subunit Ribosomal DNA Sequences Research Articles

Molecular phylogeny of the Lecudinoidea (Apicomplexa): A major group of marine gregarines with diverse shapes, movements and hosts.

Gregarine apicomplexans are ubiquitous endosymbionts of invertebrate hosts. Despite their ecological and evolutionary importance, inferences about the phylogenetic relationships of major gregarine groups, such as the Lecudinidae and Urosporidae, have been hindered by vague taxonomic definitions and limited molecular and morphological data. In this study, we investigated five gregarine species collected from four families of polychaete hosts (Nereididae, Oenonidae, Hesionidae, and Phyllodocidae) using light microscopy (LM) and scanning electron microscopy (SEM). We also generated small subunit ribosomal DNA sequences from these species and conducted molecular phylogenetic analyses to elucidate the evolutionary relationships within the Lecudinoidea. Our results include new molecular and morphological data for two previously described species (Lecudina cf. platynereidis and Lecudina cf. arabellae), the discovery of a new species of Lecudina (L. oxydromus n. sp.), and the discovery of two novel species, namely Amplectina cordis n. gen. et. n. sp. and Sphinctocystis inclina n. sp. These two species exhibited unique shapes and movements, resembling those of urosporids but with a phylogenetic affinity to lecudinids, blurring the border between lecudinids and urosporids. Our study emphasizes the need for further investigations into this highly diverse group, which has achieved great success across multiple animal phyla with diverse shapes and movements.

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.

Exotic myxozoan parasites establish complex life cycles in farm pond aquaculture

Myxozoans are obligate parasites with complex life cycles, typically infecting fish and annelids. Here, we examined annelids from fish farm pond sediments in the Beit Shean Valley, in the Syrian-African Rift Valley, Israel, for myxozoan infections. We examined 1486 oligochaetes, and found 74 (5 %) were infected with actinospore stages. We used mitochondrial 16S sequencing to infer identity of 25 infected annelids as species of Potamothrix, Psammoryctides, Tubifex and Dero. We identified 7 myxozoan types from collective groups Neoactinomyxum and Sphaeractinomyxon, and characterized them by small subunit ribosomal DNA sequencing. The Neoactinomyxum type was genetically most similar (∼93 %) to cyprinid fish-infecting Myxobolus spp. The six Sphaeractinomyxon types were genetically similar (93–100 %) to Mugilid-infecting Myxobolus spp.; with one being the previously unknown actinospore stage of a myxospore that infects mullet from aquaculture from the Israeli coast of the Mediterranean Sea. As the farm pond system is artificial and geographically isolated from the Mediterranean, the presence of at least seven myxozoans in their annelid hosts demonstrates introduction and establishment of these parasites in a novel, brackish environment.

First report of Myxobolus neurofontinalis (Bivalvulida: Myxobolidae) infecting anadromous Brook Trout from Prince Edward Island, Canada.

During routine histological examination of tissues from mortality events of anadromous Brook Trout Salvelinus fontinalis from Prince Edward Island (PEI), Canada, myxospores consistent with Myxobolus were observed infecting the central nervous system. The objective of this study was to identify the species of Myxobolus infecting the nervous system of anadromous Brook Trout from PEI, Canada. Myxospore morphology, small subunit (SSU) ribosomal DNA (rDNA) sequence data, and histology were used to identify myxospores isolated from infected Brook Trout. Myxospore measurements from the PEI samples matched those reported in the description of Myxobolus neurofontinalis from North Carolina. A 1057-bp fragment of the SSU rDNA from myxospores collected from Brook Trout in PEI was identical to an isolate of M. neurofontinalis (MN191598) collected previously from the type locality, New River basin, North Carolina. Histological sections confirmed infections were intercellular in the central nervous system. Minimal host response was observed, with only sparse mononuclear inflammatory infiltrates present at the periphery of and within dispersed myxospores, suggesting that infections are not pathogenic to Brook Trout. Myxospores were identified as M. neurofontinalis, which was previously described from the central nervous system of Brook Trout from the New River basin, North Carolina, USA. This constitutes the first time M. neurofontinalis has been documented outside of the New River basin in North Carolina.

Molecular characterisation and phylogeny of two nematode parasites recovered from the amphibian host Schlerophrys regularis sampled in Lagos metropolis, Nigeria

Research on amphibian parasites has been on the increase because of the noticeable decline of amphibians. Studies on these parasites have been limited to their prevalence and infection patterns, and there is still a dearth of knowledge on the phenotypic variations. To characterise and investigate the genetic variability and relatedness of amphibian nematode parasites, two nematode parasites–suspected Cosmocerca sp. and suspected Rhabdias sp.–of the toad Schlerophrys regularis were investigated. One hundred and sixty-eight S. regularis were collected from three study areas: Ikorodu, Ojota and Badagry. Intestines were dissected to obtain the enteric parasites, and lungs were washed in saline solution. Parasites were labelled according to the location of collection (Badagry 5A-5I, Ikorodu 5J–5O, Ojota 5P and 5S) and were then preserved for further identification. Eighteen parasites were identified morphologically using standard keys and by amplification of the 18S small subunit (SSU) ribosomal DNA sequence. Phylogenetic analysis was done using the Neighbour-Joining method. From the sequence homology and phylogenetic analysis, the suspected Cosmocerca sp. shared the closest genetic relatedness with Aplectana chamaeleonis isolate AC3 with 100% similarities while the suspected Rhabdias sp. shared the closest genetic relatedness with Rhabditoides regina isolate DF5012 with 100% similarities. Based on location, four Aplectana chamaeleonis (5E, 5F, 5G, 5H) and five Rhabditoides regina (5A, 5B, 5C, 5D, 5I) were from Badagry, four Aplectana chamaeleonis (5J, 5M, 5N, 5O) and two Rhabditoides regina (5K, 5L) from Ikorodu, one Aplectana chamaeleonis (5S) and two Rhabditoides regina (5P, 5Q) from Ojota. We report that Rhabditoides regina, a parasite known to be associated with insects, was recovered in the lungs of toads. Further investigations to resolve the complexities of superfamily Cosmocercoidea are recommended. Relatedness was not influenced by collection location as parasites were not unique to a specific environment.

Morphological and molecular characterization of a new freshwater Ceratomyxa species (Cnidaria: Myxozoa) from the yellow catfish, Trachysurus fulvidraco in China.

Ceratomyxa Thélohan, 1892 is one of the largest genera of the sub-phylum Myxozoa, and has a worldwide geographical distribution. Ceratomyxa species mainly infect the gallbladder of marine fish and rarely infect histozoically or coelozoically freshwater fish. In the present study, yellow catfish, Trachysurus fulvidraco (Siluriformes, Actinopteri) collected from the low reach of Yellow River was first found to be infected with an unknown Ceratomyxa species in their gallbladder which was identified to be new to science and nominated as Ceratomyxa huangheensis n. sp. by an integrative taxonomic approach for myxosporeans. Spores are typical of the genus Ceratomyxa, and matures spores are arcuate, with posterior angle of 139.2±11.6 (137.0-156.0)° and rounded ends, and measures 4.7±0.6 (3.3-5.5) μm in length, and 10.7±1.3 (8.5-13.3) μm in thickness. Two spore valves are slightly unequal and present elongated ovoid in the lateral view. Two equal spherical polar capsules, measuring 2.2±0.4 (1.4-2.8) μm×2.0±0.4 (1.0-2.5) μm in size locates adjacent to the straight suture line. The obtained partial small subunit ribosomal DNA (SSU rDNA) sequence was unique among all myxozoans in GenBank, and the highest similarity is 85.3% with Ceratomyxa sparusaurati, a marine fish-infecting congener. Phylogenetic analysis further indicated that this novel species did not cluster with other freshwater fish-infecting congeners from South America to form an independent clade, but was phylogenetically positioned within the marine fish-infecting clade. The possible presence of different radiation trajectories between Ceratomyxa huangheensis n. sp. and Amazonian freshwater Ceratomyxa lineage was discussed. This is the first report of Ceratomyxa species in the Yellow River basin and the second freshwater Ceraotomyxa species in China, even in Asia.

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.

Myxobolus xiushanensis n. sp. (Myxozoa: Myxobolidae) infecting the gill arches of Yellowhead Catfish Tachysurus fulvidraco from China.

Myxosporidiosis of bagrid fishes has been a focus of aquaculture research in recent years. The purpose of this study is to characterize a novel myxobolid, named Myxobolus xiushanensis n. sp., infecting Yellowhead Catfish Tachysurus fulvidraco in China. We used molecular biology, morphology, phylogeny, and histopathology in the present study. Mature myxospores were circular to ellipsoidal in valve view, measuring 12.2 ± 0.4 μm (mean ± SD; range = 11.2-13.2 μm) in length and 10.6 ± 0.4 μm (9.5-11.1 μm) in width. Two oval polar capsules were equal in width (3.4 ± 0.2 μm; 3.0-3.8 μm) but slightly unequal in length: 5.6 ± 0.3 μm (5.3-6.1 μm) and 4.7 ± 0.2 μm (4.4-5.5 μm). The polar capsule was packed with five to seven spirals of polar tubules. Histopathological investigation demonstrated that the plasmodium under the cuticular layer of the gill arch only induced a local inflammatory response and did not cause serious damage to the gill arch's internal structure. The two small subunit (SSU) ribosomal DNA sequences of M. xiushanensis n. sp. showed 100% similarity and uniqueness, and the highest similarity with other myxosporean sequences in GenBank was 90.27% (query coverage = 94%). The secondary structures of the SSU ribosomal RNA revealed that the present species was distinctly different from related species in regions V4 and V7. Phylogenetic analysis showed that M. xiushanensis n. sp. clustered independently within a branch. These results enrich our understanding of the biodiversity of myxobolids infecting bagrid fishes and provide fundamental data for the diagnosis of myxosporidiosis.

A new species of Ellipsomyxa Køie, 2003 (Bivalvulida) infecting the gall bladder of Pangasius macronema Bleeker (Siluriformes: Pangasiidae) from the Mekong River Delta, Vietnam.

During a parasitological survey of freshwater fishes in the Mekong River Delta, Vietnam, disporic plasmodia containing myxospores morphologically consistent with Ellipsomyxa Køie, 2003 (Bivalvulida) were observed infecting the gall bladder of Pangasius macronema Bleeker (Siluriformes: Pangasiidae). Herein, we use morphology and small subunit ribosomal DNA (SSU rDNA, 18S) sequence data to describe Ellipsomyxa intravesica Ksepka & Bullard n. sp. and relate it to other myxozoans. The new species resembles Ellipsomyxa adlardi Whipps & Font, 2013, which infects the naked goby, Gobiosoma bosc (Lacepede) (Gobiiformes: Gobiidae) in Lake Pontchartrain, Louisiana, but differs from it by having a longer myxospore (mean = 13.3; range = 12.0-15.0 vs. 12.4; 11.3-14.4) and shorter polar capsules (3.7; 3.0-4.0 vs. 4.3; 3.9-4.9). The 18S phylogenetic analysis recovered the sequence of the new species sister to those ascribed to Ellipsomyxa ariusi Chandran, Zacharia, Sathianandan & Sanil, 2020 and Ellipsomyxa sp. (MK561979); both of which infect the gall bladder of the threadfin sea catfish, Arius arius (Hamilton) (Siluriformes: Ariidiae) from the southwest coast of India. Consistent with previous phylogenetic analyses of Ellipsomyxa spp., Ellipsomyxa was recovered as monophyletic. The new species is the first species of Ellipsomyxa reported from a freshwater fish in Asia and the first myxozoan reported from P. macronema.

Molecular characterization of Blastocystis and Entamoeba of muskoxen and sheep in Greenland

Molecular characterisation of endobionts that are shared among human and non-human hosts can help shed light on the epidemiology and inform studies that aim to unravel the role of these organisms in health and disease. Two of the most common of shared endobionts include the single-celled intestinal protists Blastocystis and Entamoeba. Here, we present the first known data on genetic diversity and host specificity of these two genera in Greenland. Faecal DNA samples from 243 muskoxen and 44 sheep were submitted to metabarcoding of nuclear small subunit ribosomal DNA. Entamoeba- and Blastocystis-specific sequences were clustered, and consensus sequences were subjected to taxonomic query. Using MinION-based sequencing, near-complete nuclear small subunit ribosomal DNA sequences were obtained from four faecal samples. Of the 243 muskox samples, 180 (74%) and 19 (8%) were positive for Blastocystis and Entamoeba, respectively. Forty (91%) and six (14%) of the 44 sheep samples were positive for Blastocystis and Entamoeba, respectively. Blastocystis subtypes (ST) 10, 14, 21, 24–26, and a novel subtype (ST40) were identified. Colonisation by more than one subtype was common. ST40 was common in muskoxen but limited to Northeast Greenland. Entamoeba bovis and the E. bovis-associated ribosomal lineages (RL) 1 and 8 were found, and three conditional lineages (CL) 3, 4, and 10 were confirmed; CL10 was promoted to RL12. Several novel lineages were identified, all of which were linked to the E. bovis complex. In conclusion, Blastocystis was far more common than Entamoeba and found in approximately three of every four animals; both can be considered common colonisers of large herbivorous mammals in Greenland. Multiple subtypes/lineages of both genera were commonly observed, some of which were novel, but most of which are seen in many other parts of the world.

Henneguya correai n. sp. (Cnidaria, Myxozoa) Parasitizing the Fins of the Amazonian Fish Semaprochilodus insignis

We used a combination of morphological, molecular and biological data to characterize a novel Henneguya (Myxozoa) species infecting the Amazonian prochilodontid Semaprochilodus insignis or “kissing prochilodus”, a popular food fish and aquarium species in the Brazilian Amazon. Twenty-one S. insignis were caught live from the Tapajós river, Pará State, Brazil, then examined for myxozoan infections. Cysts of a novel Henneguya species were observed in the connective tissue of the fins. Myxospores measured 48 ± 4.9 (39.5–60.8) µm total length, of which caudal appendages were 33 ± 4.5 (26.4–45.2) µm and spore body was 15 ± 1.6 (12.4–20.5) µm. The spore body was 4.0 ± 0.6 (2.7–5.3) µm wide × 3.2 ± 0.4 (2.7–3.6) µm thick, with two unequal polar capsules (nematocysts) 7.2 ±0.8 (5.2–8.3) × 1.5 ± 0.3 (1.0–2.2) µm for the larger capsule and 5 ± 0.7 (4.0–6.3) × 1.4 ± 0.2 (1.0–1.8) µm for the smaller capsule. Polar tubules had 8–13 turns. Generative cells, immature and mature myxospores were observed within plasmodia. Ultrastructure showed plasmodia surrounded by collagen fibers, with the plasmodial membrane having pinocytotic channels. Phylogenetic analysis of small subunit ribosomal DNA sequences showed that the new Henneguya species clustered as a sister taxon to Henneguya tietensis, a parasite of the gills of the prochilodontid fish Prochilodus lineatus, from the geographically distant Paraná–Paraguai River basin.

Isolation and identification of an arbuscular mycorrhizal fungus specifically associated with mycoheterotrophic seedlings of Gentiana zollingeri (Gentianaceae).

Gentiana zollingeri (Gentianaceae) is an initial mycoheterotrophic plant that depends on a specific group of arbuscular mycorrhizal (AM) fungi for carbon source during underground growth after seed germination. In this study, a mycorrhizal fungus dominant in mycoheterotrophic seedlings of G. zollingeri was successfully isolated from a soil core collected from a point close to a flowering G. zollingeri. The AM fungal isolate was identified as conspecific or closely related to Dominikia aurea (Glomeraceae) by spore morphology and molecular phylogeny. Basic Local Alignment Search Tool (BLAST) searches against the MaarjAM database showed that the nuclear small subunit ribosomal DNA sequences of the isolate matched the AM fungal sequences obtained from a wide range of plants in various ecosystems, including several mycoheterotrophs. Thus, it is suggested that the AM fungal isolate is one of the cheating susceptible AM fungi. Furthermore, the sequences corresponded to those of a group of AM fungi dominantly detected in Japanese temperate forests. Accordingly, there is a possibility that mycoheterotrophic plants, including seedlings of G. zollingeri, may target AM fungi with a wide host range and ubiquitous distribution.

Morphological Characterization of Fresh and 20-Yr-Old Fixed Nematode Specimens of Sauertylenchus Maximus (Allen, 1955) Siddiqi, 2000 Deposited in the USDA Nematode Collection from Arlington National Cemetery, VA, USA.

Sauertylenchus maximus was discovered during a survey conducted at the Arlington National Cemetery, Virginia, for the type specimens of Hoplolaimus galeatus. Besides the fresh material, the fixed specimens of S. maximus were also studied by molecular and morphological means. The morphological and morphometric characteristics of the recovered fresh material were consistent with the original and other description(s) of this species. The fixed specimens used in this study were preserved in a 3% formaldehyde and 2% glycerin solution for over 20 yr. Molecular analyses of the fresh and fixed specimens were performed using internal transcribed spacer, D2-D2 expansion segments of 28S large subunits, and 18S small subunit ribosomal DNA sequences. To our knowledge, this represents the first report of S. maximus from Virginia and the first report of a successful DNA extraction from fixed nematode specimens.

Further insight into the genetic diversity of Entamoeba coli and Entamoeba hartmanni.

Despite the species' wide distribution, studies of the genetic diversity within Entamoeba coli and Entamoeba hartmanni remain limited. In the present study, we provide further insight into the genetic diversity of both species based on analysis of partial nuclear small subunit ribosomal DNA sequences generated from human fecal DNAs from samples collected in Africa, South America, and Europe. Reinforcing the previous recognition that E. coli is a species complex, our data confirm the existence of the two subtypes, ST1 and ST2, previously identified plus, potentially, a new subtype, ST3. While ST1 appears to be genetically quite homogenous, ST2 shows a substantial degree of intrasubtype diversity. ST2 was more common in samples collected outside Europe, whereas ST1 showed no geographical restriction. The potentially novel subtype is represented to date exclusively by sequences from South American and African samples. In contrast to previous reports, our new data also indicate substantial variation in E. hartmanni that could also support the establishment of subtypes within this species. Here, however, no links were identified between subtype and geographical origin.

The taxonomy and molecular phylogeny of two epibiotic colonial peritrich ciliates (Ciliophora, Peritrichia)

Two colonial sessilid peritrichs, Epistylis qingdaoensis n. sp. and Carchesium cyclopidarum Nenninger, 1948, were isolated from a marine crustacean and a freshwater mayfly, respectively. Morphological characters for each species were revealed by in vivo observations and silver staining methods. Epistylis qingdaoensis n. sp. is characterized by the apperance of the colony which is up to 250 μm high and usually contains fewer than eight zooids, the single-layered peristomial lip, the conspicuously conical peristomial disc, and the structure of infundibular polykinety 3 which comprises three isometric ciliary rows. Carchesium cyclopidarum is recognized by the colony height of about 600 μm, the small zooid size of 35–50 × 20–30 μm, and the low number of silverlines. Phylogenetic analyses based on small subunit ribosomal DNA sequences were performed to reveal their evolutionary relationships. Surprisingly, neither species clustered with its congeners. Epistylis qingdaoensis n. sp. nested within a clade of Zoothamnium species that was sister to a clade comprising core Epistylis species and several Zoothamnium species. Carchesium cyclopidarum nested within a clade comprising solitary peritrichs and was distantly related to the type species of Carchesium, C. polypinum. These findings imply for the first time that Carchesium is non-monophyletic.

A novel Ceratomyxa species (Myxozoa: Cnidaria) infecting an Amazonian catfish.

Parasites are important organisms for the health of ecosystems. While the Amazon Basin is home to a great diversity of ichthyofauna, our knowledge of myxozoan diversity in the biome remains relatively limited. The present study describes a new myxozoan species, Ceratomyxa mandii n. sp., parasitizing the gallbladder of the Amazonian catfish Pimelodina flavipinnis (Pimelodidae) from the Solimões River, in the region of Manaus, Brazil. Light and electron microscopy, small subunit ribosomal DNA (SSU rDNA) sequencing and phylogenetic analysis were performed. The new species exhibited worm-like plasmodia with undulatory motility. The SSU rDNA based phylogenetic analysis revealed it to be a sister taxon of C. gracillima, which also parasitizes an Amazonian pimelodid fish, possibly reflecting a host-parasite co-speciation process. This study contributes to our understanding of this little sampled group of organisms.

Taxonomy and Systematics of Two New Species of Myxobolus (Cnidaria: Myxobolidae) Parasitizing the Gills of Mugil curema (Mugilidae) from the Brazilian Coast

We describe two new species of Myxobolus (Myxobolidae) found parasitizing Mugil curema from two locations in Brazil: Myxobolus curemae n. sp. from gill arch and rays and Myxobolus maceioensis n. sp. from gill lamellae. We based the descriptions on myxospore morphology, histology, and small-subunit ribosomal DNA sequences. Myxospores of the two new species had similar morphology and morphometry but differed in spore body width and length, and ssrDNA sequences differed by 10.5%. These data supported the diagnosis of the parasites as distinct and novel species. The phylogenetic analysis showed a subclade formed by species that parasitize Mugiliformes, with M. maceioensis n. sp. as a sister species of Myxobolus episquamalis and Myxobolus bizerti, while there is a group of six species that are sister related to M. curemae n. sp. Our analysis was consistent with previous studies suggesting that orders of the hosts are strongly correlated with phylogenetic signals in the Myxobolidae. Myxobolus curemae n. sp. and M. maceioensis n. sp. are new species identified parasitizing M. curema.

Nucleospora hippocampi n. sp., an Intranuclear Microsporidian Infecting the Seahorse Hippocampus erectus From China

The life cycle, ultrastructure, and molecular phylogeny of a new intranuclear microsporidian, Nucleospora hippocampi n. sp., infecting the intestine of the Hippocampus erectus, were described. The histopathology revealed an extensive infection, mainly in the columnar epithelium of the intestinal mucosa layer. The enterocytes were the important target cell for Nucleospora hippocampi n. sp. infection. Transmission electron microscopy results showed that this microsporidian developed directly within the host cell nucleoplasm. In the intranuclear life cycle, the transformation from meront to sporogonial plasmodium was recognized by forming electron-dense disc structures, which were considered the polar tube precursors. The microsporidian showed the typical morphological characteristics of the family Enterocytozoonidae in the formation and development of spore organelles prior to the division of the sporogonial plasmodium. According to wet smear observation, eight spores were generally formed in a single host nucleus. Mature spores were elongated ovoids that were slightly bent and measured 1.93 × 0.97 μm. The isofilar polar tube was arranged in 7~8 coils in one row. Phylogenetic analysis of its small subunit ribosomal DNA sequences demonstrated that the parasite belonged to the Nucleospora group clade. The histological, ultrastructural, and molecular data support the emergence of a new species in the genus Nucleospora. This is the first report of Nucleospora species in Asia and threatened syngnathid fishes.

Morphological and molecular studies on two myxosporean infections of cyprinid fishes: Thelohanellus pyriformis from tench and Thelohanellus cf. fuhrmanni from nase.

During surveys on myxosporean parasites of Lake Balaton and River Danube fishes, two Thelohanellus spp. were found on tench (Tinca tinca) and on common nase (Chondrostoma nasus). They were identified as Thelohanellus pyriformis and Thelohanellus cf. fuhrmanni, respectively. Myxospores of T. pyriformis from tench were collected from arteria branchialis afferens of gill filaments. The mature myxospores of this species were pyriform in shape and 19 ± 0.6l (18–19.5) long, 8.2 ± 0.54 (7.5–9) wide, 7.3 ± 0.25 (7–7.5) thick containing polar tubules with 9–10 turns. The plasmodia of Thelohanellus cf. fuhrmanni were collected from under the skin of snout of the common nase. The myxospores were pyriform, 16.3 ± 0.39l (15.5–16.5) long, 6.5 ± 0.55 (6.3–7) wide, 6.3 ± 0.53 (5.8–7) thick containing polar tubules with 6 turns. Small subunit ribosomal DNA sequences of both Thelohanellus species differed from other known myxozoans. The myxospores morphology, histopathology and ssrDNA sequences supported a diagnosis of T. pyriformis from tench and T. cf. fuhrmanni from common nase.

Redescription and molecular characterization of two Trichodina species (Ciliophora, Peritrichia, Mobilida) from freshwater fish in China.

During an investigation of parasitic ciliates in northern China, two Trichodina species, T. acuta Lom, 1970 and T. nigra Lom, 1960, were isolated from the freshwater fish Cyprinus carpio Linnaeus, 1758. The morphology of each species was investigated based on dry silver nitrate-stained specimens. In addition, the molecular phylogeny of each was analyzed based on small subunit ribosomal DNA (SSU rDNA) sequence data. Trichodina acuta can be distinguished from its congeners by the undefined periphery of the central circle, the distinct gap between the rays and the central circle, and the distinctly sickle-shaped blades. Trichodina nigra is a cosmopolitan ciliate and is characterized by its densely linked denticles, broad, rounded spatula-shaped blades, robust central parts, and well developed rays. Phylogenetic analyses revealed that T. acuta and T. nigra nest within different clades, supporting the assertion that the GC content of SSU rDNA sequences could reflect evolutionary relationships among Trichodina species.