Small Subunit Ribosomal DNA Sequences Research Articles

Blastocystis subtypes detected in long-tailed macaques in Thailand—Further evidence of cryptic host specificity

Blastocystis is an enteric parasitic protist with a widespread distribution in a variety of human and non-human hosts and supposedly a source of zoonotic transmission. This study aimed to determine the frequency and distribution of Blastocystis subtypes in free-ranging Macaca fascicularis (long-tailed macaques, LTMs) in Thailand. A total of 628 faecal samples were collected from free-ranging LTMs inhabiting fourteen tourist attraction sites in Thailand. Fresh faecal samples were individually cultured in Jones’ medium, and Blastocystis-positive samples were subtyped using nuclear small subunit ribosomal DNA (SSU rDNA) sequencing. Two hundred and sixty-three (41.87%) samples were positive by culture, and 197 and 154 were successfully SSU rDNA-amplified and sequenced, respectively. Three subtypes (ST1, ST2, and ST3) comprising 19 alleles were observed. ST3 was the most common subtype detected (36.55%), followed by ST2 and ST1 (24.37% and 17.26%, respectively). Some subtype alleles not previously observed were identified. A couple of strains appeared similar to those found in humans as evidenced by SSU rDNA allelic profiles, while further evidence of cryptic host specificity emerged. This study provides the first data on Blastocystis subtypes in non-human primates in Thailand and confirms the trend observed in other Old-World countries with regard to the colonization rate and subtype distribution.

Dematipyriforma aquilariagen. et sp. nov., a New Hyphomycetous Taxon fromAquilaria crassna

A hyphomycetous fungus producing dark muriform conidia, was isolated as endophyte from trunk of Aquilaria crassna in Laos. The fungus was compared to morphologically similar genera, such as Annellophorella, Berkleasmium, Canalisporium, and Monodictys. Morphological evidence, however, excluded the fungal taxon from any of these genera. Phylogenetic analysis of combined small subunit, 5.8S, and large subunit ribosomal DNA sequence data placed the fungus within the order Savoryellales (Sordariomycetes, Hypocreomycetidae), but on a distinct terminal clade along with other groups of the order. Based on morphological and molecular phylogenetic evidence, we therefore established the genus Dematipyriforma to accommodate the taxon, which was named Dematipyriforma aquilaria.

Small-subunit ribosomal DNA sequencing analysis of dietary shifts during gonad maturation in wild black Amur bream (Megalobrama terminalis) in the lower reaches of the Pearl River

Black Amur bream (Megalobrama terminalis) is an economically important indigenous species that dominates the fish community in the lower reaches of the Pearl river. We used small subunit ribosomal DNA (18S rDNA) sequencing to assess the intestinal contents of black Amur bream at four independent developmental stages (stages II, III, IV, and V). This represents a new approach to the analysis of fish diets in the field. The results showed that Eunicida, Tubificina, unclassified Demospongiae, unclassified Insecta, unclassified Hexactinellida, unclassified Bivalvia and Monogononta constitute the predominant primary food sources of black Amur bream. Interestingly, a shift in the diet of black Amur bream was observed upon the fish reaching a standard length of around 225 ± 19.3 mm during gonad development from stage III to IV. A reduction in the consumption of Demospongiae and Rotifera, and an increase in the consumption of Annelida (mostly Eunicida) and Tubificina, were observed. Stage III in the developmental process is a transition phase in relation to dietary shift, which exhibits the food preferences both of juveniles and adults. We also observed that food items that were detected in the foregut were also present in the hindgut, indicating that 18S rDNA amplification of fecal matter could be used for dietary analysis in fish in the field.

Morphology, morphogenesis and molecular phylogeny of a new soil ciliate Paragonostomoides xianicum n. sp. (Ciliophora, Hypotrichia, Gonostomatidae)

A hypotrichous ciliate, Paragonostomoides xianicum n. sp., was discovered in soil on the surface of the Xi’an Circumvallation, Xi'an, China. Its morphology and some major ontogenetic stages were studied and the phylogenetic position was estimated using standard methods Paragonostomoides xianicum n. sp. is characterized as follows: body size about 75–90×15–25μm in vivo, elliptical and flexible; colourless mitochondrion-like cortical granules scattered throughout cell surface, about 0.8×0.6μm in size; contractile vacuole positioned at mid-body; frontoventral row IV extends beyond buccal vertex and frontoventral row VI extends to about body end; frontoventral cirri not in pairs; transverse cirri absent; three caudal cirri; two macronuclear nodules and one to four micronuclei. The ontogenetic process in P. xianicum n. sp. is basically similar to that in Metagonostomum gonostomoidum and Gonostomum-species and featured by: frontoventral-transverse cirri develop in five-anlage pattern, the rearmost cirri generated in the two rightmost FV anlagen are not set off posteriorly (like an ordinary TC), as well as FVT-anlagen n and n-1 form in secondary mode in P. xianicum. Phylogenetic analyses based on small subunit ribosomal DNA sequence data reveal a close relationship of P. xianicum n. sp. with Cotterillia bromelicola and two Gonostomum-species.

Ultrastructural and immunocytochemical investigation of paramylon combined with new 18S rDNA-based secondary structure analysis clarifies phylogenetic affiliation of Entosiphon sulcatum (Euglenida: Euglenozoa)

The phylogeny of phagotrophic euglenids is widely based on nuclear small subunit ribosomal DNA (18S rDNA) sequence data, but most analyses suffer from weakness in statistical support regarding the “connecting backbone” between monophyletic clades. Moreover, the position of Entosiphon has remained unclear. Testing the 18S rDNA capability for a phylogeny of phagotrophic euglenids, we isolated sequences of Peranema sp. and Ploeotia edaphica and utilized secondary structure data as a prerequisite for recognition of homologous positions. We found a unique, clade-specific nucleotide substitution in the deduced 18S rDNA helix 44. Since our 18S rDNA phylogenies could only in part resolve positions of phagotrophic lineages, but did not verify that of Entosiphon, we investigated the phagotrophic key taxa Peranema trichophorum, Petalomonas cantuscygni, Ploeotia costata, and Entosiphon sulcatum ultrastructurally. Additionally, we explored the presence or absence of the euglenid reserve carbohydrate paramylon by specific staining with monoclonal anti-β-1,3-glucan antibodies. Paramylon was found to be clearly present in P. trichophorum and E. sulcatum, but was absent in Pt. cantuscygni and Pl. costata. Combined results of our molecular, ultrastructural, and immunocytochemical investigations suggest that Entosiphon sulcatum is the sister taxon of a monophyletic euglenid crown clade, characterized by a helical pellicle, which we propose to rename. This phylogenetic affiliation is confirmed by a clade-specific primary absence of the unique nucleotide substitution in helix 44 and by the common presence of paramylon.

Amazonian waters harbour an ancient freshwater Ceratomyxa lineage (Cnidaria: Myxosporea)

A new species of Ceratomyxa parasitizing the gall bladder of Cichla monoculus, an endemic cichlid fish from the Amazon basin in Brazil, is described using morphological and molecular data. In the bile, both immature and mature myxospores were found floating freely or inside elongated plasmodia: length 304 (196–402) μm and width 35.7 (18.3–55.1) μm. Mature spores were elongated and only slightly crescent-shaped in frontal view with a prominent sutural line between two valve cells, which had rounded ends. Measurements of formalin-fixed myxospores: length 6.3±0.6 (5.1–7.5) μm, thickness 41.2±2.9 (37.1–47.6) μm, posterior angle 147°. Lateral projections slightly asymmetric, with lengths 19.3±1.4μm and 20.5±1.3μm. Two ovoid, equal size polar capsules, length 2.6±0.3 (2–3.3) μm, width 2.5±0.4 (1.8–3.7) μm, located adjacent to the suture and containing polar filaments with 3–4 turns. The small subunit ribosomal DNA sequence of 1605 nt was no more than 97% similar to any other sequence in GenBank, and together with the host, locality and morphometric data, supports diagnosis of the parasite as a new species, Ceratomyxa brasiliensis n. sp. Maximum parsimony and maximum likelihood analyses showed that C. brasiliensis n. sp. clustered within the marine Ceratomyxa clade, but was in a basally divergent lineage with two other freshwater species from the Amazon basin. Our results are consistent with previous studies that show Ceratomyxa species can cluster according to both geography and host ecotype, and that the few known freshwater species diverged from marine cousins relatively early in evolution of the genus, possibly driven by marine incursions into riverine environments.

Myxobolus taibaiensis sp. n. (Myxozoa: Myxosporea) infecting the intestinal wall of common carp Cyprinus carpio Linnaeus in China.

Myxobolus taibaiensis sp. n. was found in the inner intestinal wall of common carp, Cyprinus carpio Linnaeus, during the investigation of fish parasite fauna in Lake Taibai, located in the middle reach of the Yangtze River, China. The whitish ellipsoidal plasmodia, up to 2.9 mm long and 1.7 mm wide, developed in the circular muscle layer of the intestinal wall and produced significant compression into adjacent tissues, but no significant inflammatory responses were observed against this infection. Mature spores are oval in frontal view and lemon-like in lateral and apical view, averaging 10.2-11.2 µm (10.8 ± 0.2 µm) in length, 9.1-9.9 µm (9.6 ± 0.2 µm) in width and 6.1-6.6 µm (6.3 ± 0.1 µm) in thickness. Polar capsules are pyriform, equal in size, slightly converging anteriorly, measuring 4.4-5.4 µm (5.0 ± 0.2 µm) in length by 3.2-3.6 µm (3.4 ± 0.1 µm) in width. Polar filaments coiled with four to five turns and arranged perpendicular to the polar capsule length, measuring up to 106 µm. Myxobolus taibaiensis sp. n. is morphologically similar to Myxobolus rotundatus Achmerov, 1956 which also infects the inner wall of the intestine of common carp. However, the small subunit ribosomal DNA sequence identity was only 94%, generally beyond the intraspecies variation in the genus. Phylogenetically, this new species is sister to M. rotundatus and then clusters with M. shantungensis Hu, 1965 to form an independent common carp-infecting cluster within the Henneguya-Myxobolus clade.

A novel myxosporean parasite Myxobolus klamathellus n. sp. (Cnidaria: Myxosporea) from native blue chub (Gila coerulea) in Klamath Lake, Oregon

Blue chub, Gila coerulea Girard, 1856 is a freshwater cyprinid fish native to inland drainages of western North America. It has not previously been recorded as a host of any myxosporean parasite (Cnidaria: Myxosporea), despite myxosporeans being cosmopolitan in freshwater and marine fishes worldwide and sympatric with this host. Herein, we describe a novel myxosporean from subcutaneous cysts in native blue chub from Klamath Lake, Oregon. Myxospores were consistent with genus Myxobolus, being obovoid but compressed in thickness, length 14.3 ± 0.4 (13-15) μm, width 9.7 ± 0.4 (9-10) μm, thickness 7.7 ± 0.3 (7-8) μm; two polar capsules ovoid slightly dissimilar in size, length 6.4 ± 0.4 (6-7) μm, width 3.8 ± 0.3 (3-4) μm, with four (3-5) turns of the polar filament (tubule); capsule openings apical, one in each valve cell. The small subunit ribosomal DNA sequence was up to 97% similar to Myxobolus spp. from other cyprinids from North America and Europe. Given the novel host, unique myxospore morphometrics, and DNA sequence, we describe this as Myxobolus klamathellus n. sp.

Morphology, morphogenesis and molecular phylogeny of a novel saline soil ciliate, Lamtostyla salina n. sp. (Ciliophora, Hypotricha)

The morphology and morphogenesis of a new saline soil hypotrich, Lamtostyla salina n. sp., collected from Longfeng Wetland in Daqing, north China, were studied based on live observations and protargol stained specimens. The new species is characterized as follows: body very flexible but not contractile, lanceolate with anterior end broadly rounded, widest at about 1/3 of body length, posterior end narrowly rounded; cortical granules colourless and scattered; amphisiellid median cirral row ends ahead of mid-body, composed of 5–13 cirri; 4–20 frontoventral cirri arranged in 2–4 rows; three frontal, one buccal and 2–6 transverse cirri; usually one left and one right marginal row, composed of 17–51 and 20–51 cirri respectively; usually two dorsal kineties; two macronuclear nodules and two micronuclei. Morphogenesis is typical of the genus Lamtostyla: parental structures are involved in the formation of frontoventral transverse anlagen in the proter. Phylogenetic analyses based on small subunit ribosomal DNA sequence data reveal that the systematic position of Lamtostyla is rather unstable with low support values across the tree. However, a well-recognized close relationship between Lamtostyla and Bistichella are shown in all phylogenetic analyses.

Morphological description and phylogeny of Ceratomyxa scorpaeni n. sp. (Myxosporea: Ceratomyxidae) infecting the gallbladder of Scorpaena porcus (L.) (Scorpaeniformes: Scorpaenidae) from the bay of Bizerte in Tunisia.

A new marine myxosporean species Ceratomyxa scorpaeni n. sp. is described using morphological characteristics and small subunit ribosomal DNA (rDNA) sequence data. The parasite was found infecting the gallbladder of Scorpaena porcus Linnaeus, 1758 collected from the Bay of Bizerte. Mature plasmodia were disporous free floating in the bile. Spores were transversely elongated and crescent-shaped with unequal shell valves and measured 7.6 ± 0.2 (7.2-8.0) μm in length and 32.8 ± 0.3 (21.6-44.0) μm in thickness. Two sub-spherical polar capsules equal in size, situated at the same level and measured 3.0 ± 0.2 (3.0-3.2) μm in length and 2.4 ± 0.3 (2.3-3.3) μm in width. The infection with C. scorpaeni n. sp. was prevalent throughout the year with a maximum prevalence of infection during spring and summer. Molecular analysis of the SSU rRNA gene confirmed the parasite as a new member of the genus Ceratomyxa and showed close relationship with the species of Ceratomyxa infecting the gallbladders of Sparidae from the Mediterranean Sea. This is the first report of Ceratomyxa species from the gallbladder of S. porcus in Tunisian coastal waters.

Morphology and small subunit rDNA-based phylogeny of Ceratomyxa amazonensis n. sp. parasite of Symphysodon discus, an ornamental freshwater fish from Amazon

The specious genus Ceratomyxa Thélodan, 1892, infect mainly gallbladder of marine fishes, with only five species reported infecting species from freshwater environment. This study performed morphological and phylogenetic analyses involving a new Ceratomyxa species (Ceratomyxa amazonensis n. sp.) found in gallbladder of Symphysodon discus Heckel, 1840 (Perciformes: Cichlidae), an important ornamental fish endemic to Amazon basin. Mature spores were strongly arcuate shaped and measured 7.0 ± 0.3 (6.2-7.6) μm in length, 15.8 ± 0.4 (15.0-16.7) μm in thickness, and polar capsules 3.22 ± 0.34 (2.4-3.6) μm in length and 2.63 ± 0.17 (2.4-2.9) μm in width. This was the first small subunit ribosomal DNA (SS rDNA)sequencing performed to Ceratomyxa species parasite of freshwater fish, and the phylogenetic analysis showed C. amazonensis n. sp. clustering in the early diverging subclade of the ceratomyxids, together with species of parasites of amphidromous/estuaries fishes, suggesting some role of the transition of the fishes between marine/freshwater environments in the evolutionary history of these parasites.

Intestinal disease of scattered mirror carp Cyprinus carpio caused by Thelohanellus kitauei and notes on the morphology and phylogeny of the myxosporean from Sichuan Province, southwest China

The mass mortality of pond-reared scattered mirror carp, Cyprinus carpio, caused by Thelohanellus kitauei, occurred at fish farms in Sichuan Province, southwest China. Morphological and molecular analyses were supplemented with histological evaluation of infected tissues to better understand the route of infection and the pathological effects of T. kitauei on the fish host. The intestine of the diseased host was full of large cysts of the myxosporean. The cysts range from 2 cm to 3.6 cm in diameter. Histopathology indicated that T. kitaue i first invaded the submucosa of the host intestine and then moved into the mucosa layers with the development of their spores, finally entering into the enteric cavity of the hosts after the disruption of mucosa layers. The pyriform spores of T. kitauei were surrounded by the transparent spore sheath, measuring 25.98 μm±0.95 μm in body length, 8.72 μm±0.51 μm in body width, and 7.86 μm±0.26 μm in body thickness. The single polar capsule was pyriform, measuring 14.73 μm±0.92 μm in length and 6.82 μm±0.45 μm in width, with eight to 10 turns of filament coils winding inside. Phylogenetic analysis based on the 18S small-subunit ribosomal DNA sequences indicated that minimal genetic differences were present between T. kitauei samples from South Korea and from China. Close affinity was found between the genus Thelohanellus and Myxobolus. Additionally, two polar capsule nuclei were found at the anterior end of the single polar capsule in spores of T. kitauei stained with hematoxylin and eosin, which suggested the separation of the genus Thelohanellus from Myxobolus.

Morphological re-description and molecular characterization of Kudoa pagrusi (Myxosporea: Multivalvulida) infecting the heart muscles of the common sea bream fish Pagrus pagrus (Perciformes: Sparidae) from the Red Sea, Egypt.

In the present study, 100 samples of different sizes of the common sea bream fish Pagrus pagrus were collected from the Egyptian water along the Gulf of Suez, Red Sea and examined for the prevalence of myxosporidian parasites in general and Kudoa spp. in particular. Fish samples were thoroughly externally examined. After dissection, all the internal organs were removed and examined. A total of 60 out of 100 fish specimens were found to be infected with Kudoa stages. Parasitic infection was restricted to the heart muscles of the examined fish. None of the other organs was found to be infected. Macroscopic cysts (plasmodia) heavily infested the different parts of the heart muscles. Each plasmodium measured 1.2-2.5 (1.53 ± 0.2) mm × 0.63-0.80 (0.65 ± 0.2) mm. Mature spores are quadratic in shape in the apical view showing four equal valves and four symmetrical polar capsules. Fresh spores were 5.0-7.1 (5.7 ± 0.2) μm long × 5.4-8.5 (6.1 ± 0.3) μm wide. On the basis of spore morphology, the present species was identified as Kudoa pagrusi. Morphometric characterization revealed that the relatively small size of this Kudoa species was the distinctive feature that separates it from all previously described species. Molecular analysis based on small subunit ribosomal DNA (SSU rDNA) sequences revealed that the highest percentage of identity was observed with K. scomberomori and followed by K. shiomitsui, K. hypoepicarclialis, K. amamiensis, and K. kenti. The kudoid spores showed morphometric variations to some extents but had essentially identical nucleotide sequences of the SSU rDNA gene sequences closest to those of K. scomberomori and K. shiomitsui recorded from elasmobranchs in the Indo-Pacific Ocean. The present findings support the identification of an ancestral marine origin of the present Kudoa species.

Supplemental description and molecular characterization of Myxobolus miyarii Kudo, 1919 (Myxosporea: Myxobolidae) infecting intestine of Amur catfish (Silurus asotus).

Myxobolus miyairii Kudo, 1919 was first reported from the intestines of the Amur catfish (Silurus asotus) in Japan and then in China and Russia, but with incomplete description. During the investigation of fish myxosporean diversity in Poyang Lake, the biggest lake along the Yangtze River, China, two Amur catfish highly infected with M. miyairii in the intestine wall were sampled. So, the complete description of this species with morphological and molecular data was presented here. A large number of whitish, round or ellipsoidal pseudocysts 0.32-0.78 mm in diameter could be found in the external intestinal wall after dissecting the infected fish. Mature spores of M. miyairii were elongated and ellipsoidal in the frontal view and narrow fusiform in the lateral view, with a slightly pointed anterior end and a bluntly rounded posterior end and measured 13.3 ± 0.49 (12.5-14.7) μm × 6.6 ± 0.27 (6.2-7.4) μm × 5.0 ± 0.26 (4.4-5.7) μm in size. Spore surface was smooth and two spore valves symmetrical, with a thin and straight sutural ridge. Interestingly, two types of caudal appendage (single or bifurcated) were occasionally present on the posterior end of some spores which has not previously been reported. The two equal pyriform polar capsules measured 6.5 ± 0.30 (6.2-7.5) μm long and 1.9 ± 0.14 (1.5-2.3) μm wide and situated at the anterior end of the spore. Polar filaments coiled with eight to nine turns, perpendicularly to the longitudinal axis of the polar capsules. Histopathological analysis showed that the plasmodium developed in the circular muscle layer of intestinal wall of Amur catfish, but no obvious inflammatory responses were observed. Phylogenetic analysis based on the partial 18S small subunit ribosomal DNA sequences indicates that M. miyairii cluster within a clade of Siluriforme-infecting Henneguya species with the support of a high bootstrap value, but also evolutionarily independent from the Henneguya clade infecting the epithelium of fish of the Ictaluridae family. Additionally, Myxobolus species reported with caudal processes dispersed within the Henneguya-Myxobolus clade.

Morphology and systematics of two freshwater urostylid ciliates, with description of a new species (Protista, Ciliophora, Hypotrichia)

The morphology of two freshwater urostylid species, Neourostylopsis flava spec. nov. and Pseudourostyla subtropicaChen et al., 2014, isolated from freshwater ponds in northern and southern China, respectively, was investigated following examination of specimens in vivo and following protargol staining. Neourostylopsis flava spec. nov. is distinguished from its congeners by the following characteristics: body size 150–220×50–75μm in vivo; yellow in colour; bright yellow to yellow-brownish spherical cortical granules densely arranged along marginal cirral rows and in irregular short rows on dorsal side; adoral zone with 40–55 membranelles; six to eight frontal, three or four buccal, two pretransverse ventral and seven to nine transverse cirri; 27–40 midventral pairs extending to about anterior 55% of cell; four or five left and four right marginal rows; freshwater habitat. A redescription of a freshwater population of P. subtropica is also provided. Phylogenetic analyses based on small subunit ribosomal DNA sequences shows that P. subtropica and N. flava spec. nov. group with their congeners and both Neourostylopsis and Pseudourostyla are monophyletic.

The molecular characterization of the enigmatic dinoflagellate Kolkwitziella acuta reveals an affinity to the Excentrica section of the genus Protoperidinium

Kolkwitziella acuta is a cyst-forming dinoflagellate with a unique tabulation, occurring in freshwater to brackish environments of Eurasia and the USA. Based on the unique thecal plate arrangement, this species was previously interpreted as a missing link in the evolution between the genus Protoperidinium and the Diplopsalioideae. We isolated living K. acuta cysts from the south-western Caspian Sea and the central Baltic (Finland), and re-established the cyst-theca relationship. The cysts had a distinctive polar and dorsoventral compression and a characteristic wrinkly texture. The plate formula of the thecate stage was Po, X, 4′, 2a, 7′′, 3c+t, ?s, 5′′′, 1′′′′. Large subunit (LSU) and small subunit (SSU) ribosomal DNA sequences obtained through single-cell PCR of these cysts reveal that this species is close to Protoperidinum excentricum which belongs to the Excentrica section of the genus Protoperidinium. We suggest that it evolved from the genus Diplopsalis through a split of the apical and anterior intercalary plates. Furthermore, that the split of the antapical plates occurred several times independently in the evolution of Protoperidinium and Diplopsalioideae and is therefore a polyphyletic trait. The cyst of Kolkwitziella is considered an indicator of low salinities, specifically between 0 and 22 psu. Geochemical analysis of the cyst walls suggests a heterotrophic mode of nutrition.

Cryptic infection of a broad taxonomic and geographic diversity of tadpoles by Perkinsea protists

The decline of amphibian populations, particularly frogs, is often cited as an example in support of the claim that Earth is undergoing its sixth mass extinction event. Amphibians seem to be particularly sensitive to emerging diseases (e.g., fungal and viral pathogens), yet the diversity and geographic distribution of infectious agents are only starting to be investigated. Recent work has linked a previously undescribed protist with mass-mortality events in the United States, in which infected frog tadpoles have an abnormally enlarged yellowish liver filled with protist cells of a presumed parasite. Phylogenetic analyses revealed that this infectious agent was affiliated with the Perkinsea: a parasitic group within the alveolates exemplified by Perkinsus sp., a "marine" protist responsible for mass-mortality events in commercial shellfish populations. Using small subunit (SSU) ribosomal DNA (rDNA) sequencing, we developed a targeted PCR protocol for preferentially sampling a clade of the Perkinsea. We tested this protocol on freshwater environmental DNA, revealing a wide diversity of Perkinsea lineages in these environments. Then, we used the same protocol to test for Perkinsea-like lineages in livers of 182 tadpoles from multiple families of frogs. We identified a distinct Perkinsea clade, encompassing a low level of SSU rDNA variation different from the lineage previously associated with tadpole mass-mortality events. Members of this clade were present in 38 tadpoles sampled from 14 distinct genera/phylogroups, from five countries across three continents. These data provide, to our knowledge, the first evidence that Perkinsea-like protists infect tadpoles across a wide taxonomic range of frogs in tropical and temperate environments, including oceanic islands.

Morphological and molecular aspects of Ceratomyxa mehlhorni n. sp., a parasite of the golden trevally Gnathanodon speciosus in the Arabian Gulf off the Saudi Arabian coast, with data on its seasonal prevalence.

During a survey of the myxosporean fauna of the golden trevally Gnathanodon speciosus from the Arabian Gulf off Saudi Arabia, a species of Ceratomyxa that did not conform to any known species was recorded. The infection was detected as a large number of mature spores free-floating in the bile. Mature spores were hat- or helm-like in the frontal view with two short and unequal valves. The spores measured 8 (7-9) × 12 (10-14). The two polar capsules were spherical, equal in size and measured 3 (2-4) in diameter with three filament turns. Sporoplasm was binucleated and filled the whole extracapsular space. The small subunit ribosomal DNA sequence of this species did not match any available sequences in GenBank. The lowest genetic distance was 0.017, observed with Ceratomyxa moseri infecting Pomacentrus wardi from Australia's Great Barrier Reef. A maximum likelihood phylogenetic tree showed a close association between the new species and a variety of ceratomyxid species, including Ceratomyxa arabica reported from the Arabian Gulf. Given the morphological and molecular differences between this species and other Ceratomyxa spp., we proposed the present form was a new species and the name Ceratomyxa mehlhorni sp. n. for this parasite from the gallbladder of G. speciosus.

Morphological, molecular and ecophysiological characterization of the phototrophic dinoflagellate Biecheleriopsis adriatica from Korean coastal waters

Biecheleriopsis is a genus of small marine planktonic dinoflagellates. Here, we present detailed morphological, molecular and ecophysiological characterizations of Biecheleriopsis adriatica based on a number of strains from Korea. The small subunit (SSU) ribosomal DNA (rDNA) sequences of two of the three Korean strains were closely related to those of previously described B. adriatica, and one was identical. However, the large subunit (LSU) and internal transcribed spacer region (ITS1, ITS2 and 5.8S) sequences from the Korean strains differed by 0.1–0.6% and 1.0–1.6%, respectively, from the non-Korean strains. In phylogenetic trees built on the basis of the SSU and LSU (D1-D3) rDNA and ITS region sequences, the Korean strains clustered within a clade including all the other B. adriatica strains. Morphologically, the Korean strains resemble those described from the Adriatic Sea. However, some cells of the Korean strains had nine latitudinal rows of amphiesmal vesicles (AVs): three episomal, three cingular, and three hyposomal rows, which is fewer than the 11 rows reported in the non-Korean strains. In addition, we report for the first time that B. adriatica has 11 sulcal plates and a small vesicle in the hyposomal ventral side. Peridinin was found to be the main accessory pigment in the Korean strains of B. adriatica. The Korean strains were not toxic to the brine shrimp Artemia salina.

Morphological examination and phylogenetic position of the newly recorded heterotrophic brackish dinoflagellate Diplopsalis caspica (Dinophyceae) in freshwater habitat from China

AbstractA heterotrophic dinoflagellate was identified as Diplopsalis caspica Ostenfeld by morphological characteristics, with the plate formula: Po, x, 3′, 1a, 6″, 4c, 4s, 5″′, 1″″. The newly recorded dinoflagellate was encountered during autumn in the phytoplankton community of Jihongtan Reservoir and some small ponds near the reservoir in Shandong Province, China. The species had been reported only in brackish ecosystems; however, our material fit with D. caspica occurred in freshwater habitat. The phylogenetic relationship of D. caspica was elucidated through small and large subunit ribosomal DNA sequences. In small and large subunit ribosomal DNA phylogenies, all Protoperidiniaceae species, including diplopsalids and Protoperidinium Bergh, could cluster into a monophyletic group. The molecular phylogenies indicated that the genus Diplopsalis would provide a link to the common ancestor of the Peridinium Ehrenberg and the Protoperidinium.