Small Subunit rDNA Sequences Research Articles

Morphological and molecular characterization of Ceratomyxa batam n. sp. (Myxozoa: Ceratomyxidae) infecting the gallbladder of the cultured Trachinotus ovatus (Perciformes: Carangidae) in Batam Island, Indonesia.

A new coelozoic myxozoan species, Ceratomyxa batam n. sp., was identified in cultured carangid fish, Trachinotus ovatus (Perciformes: Carangidae), in waters off Batam Island of Indonesia. The bi- and trivalved spores were observed in the gallbladder of T. ovatus. Mature bivalved spores of C. batam n. sp. were transversely elongated and narrowly crescent in shape, 3.8 ± 0.36 (2.7-4.6) μm long and 19.2 ± 1.75 (16.2-22.0) μm thick. Two sub-spherical polar capsules were 2.3 ± 0.18 (2.0-2.8) μm long and 2.6 ± 0.16 (2.3-2.9) μm wide. Prevalence was 72.2% in 72 examined T. ovatus according to evaluations dating from November 2016. The maximum likelihood phylogenetic tree based on small subunit rDNA sequence showed similarity with Ceratomyxa robertsthomsoni and Ceratomyxa thalassomae found in Australia. This is the first report of Ceratomyxa species identified in a seawater fish at Batam Island, Indonesia.

Chrysochromulina andersonii sp. nov. (Prymnesiophyceae), a new flagellate haptophyte symbiotic with radiolarians

ABSTRACTThe haptophyte symbiont Chrysochromulina andersonii sp. nov. isolated from the polycystine radiolarian Dictyocoryne truncatum was examined using both natural and cultured materials by light, scanning and transmission electron microscopy as well as molecular phylogenetic analyses of the small subunit (SSU) and large subunit (LSU) rDNA sequences. In culture, the motile cells were saddle-shaped, 3–5 µm long and 3–4 µm wide. They possessed two equal to subequal flagella, a coiling haptonema, and two parietal golden-brown chloroplasts. The scales were arranged in two layers composed of two types: scales with flat rims, 0.19–0.26 µm in diameter, formed the inner layer; whereas, scales with upright rims, 0.25–0.45 µm in diameter, formed the outer layer. Both scales had a faint cross in the centre of the scale face, and the radiating ridges extended from the faint cross to the periphery. Molecular phylogenetic analysis indicated that the SSU and LSU rDNA sequences from our symbiont fell within the clade of Chrysochromulina. However, our symbiont differed in the details of the patterning on the scale face from the other species of Chrysochromulina. Thus, we propose C. andersonii sp. nov. for the symbiotic alga from the polycystine radiolarian D. truncatum.

Further analyses of variation of ribosome DNA copy number and polymorphism in ciliates provide insights relevant to studies of both molecular ecology and phylogeny.

Sequence-based approaches, such as analyses of ribosome DNA (rDNA) clone libraries and high-throughput amplicon sequencing, have been used extensively to infer evolutionary relationships and elucidate the biodiversity in microbial communities. However, recent studies demonstrate both rDNA copy number variation and intra-individual (intra-genomic) sequence variation in many organisms, which challenges the application of the rDNA-based surveys. In ciliates, an ecologically important clade of microbial eukaryotes, rDNA copy number and sequence variation are rarely studied. In the present study, we estimate the intraindividual small subunit rDNA (SSU rDNA) copy number and sequence variation in a wide range of taxa covering nine classes and 18 orders of the phylum Ciliophora. Our studies reveal that: (i) intra-individual sequence variation of SSU rDNA is ubiquitous in all groups of ciliates detected and the polymorphic level varies among taxa; (ii) there is a most common version of SSU rDNA sequence in each cell that is highly predominant and may represent the germline micronuclear template; (iii) compared with the most common version, other variant sequences differ in only 1-3 nucleotides, likely generated during macronuclear (somatic) amplification; (iv) the intra-cell sequence variation is unlikely to impact phylogenetic analyses; (v) the rDNA copy number in ciliates is highly variable, ranging from 103 to 106, with the highest record in Stentor roeselii. Overall, these analyses indicate the need for careful consideration of SSU rDNA variation in analyses of the role of ciliates in ecosystems.

Morphological and molecular identification of a new Kudoa thyrsites isolate in Mediterranean silver scabbardfish Lepidopus caudatus.

Myxozoans of the genus Kudoa (Myxosporea, Multivalvulida) infect marine and estuarine fish species worldwide. Some Kudoa species are of concern to the seafood industry since they may generate macroscopic cysts in the fish host's musculature, or cause post mortem myoliquefaction, commonly known as 'soft flesh'. One of the economically most important species is K. thyrsites, a myoliquefactive myxosporean parasite that occurs in many wild and cultured marine fish species worldwide. Here we identified a K. thyrsites isolate as the causative agent of myoliquefaction in silver scabbardfish Lepidopus caudatus from the Alboran Sea (western Mediterranean Sea). For comparative and validation purposes, the morphological and molecular characteristics of the isolate were compared with fresh spores of a K. thyrsites isolate infecting Atlantic mackerel Scomber scombrus from the Norwegian Sea. Myxospores of both isolates shared a stellate appearance and contained 4 unequal pyriform polar capsules (1 large, 1 small and 2 intermediate). These morphological traits were consistent with all other previously described K. thyrsites isolates. Moreover, the small subunit rDNA sequences of the Mediterranean and Norwegian Sea isolates revealed 100% similarity, and matched 100% with K. thyrsites isolates previously recorded in myoliquefactive Atlantic mackerel from the North Sea and off southern England. The findings suggest that K. thyrsites is the primary cause of myoliquefaction in silver scabbardfish from the Alboran Sea. This report represents the first morphological and molecular characterization of K. thyrsites in the Mediterranean Sea. A set of new allometric characters is proposed as additional descriptors for more accurate and specific description of kudoid myxospores.

History of the introduction of a species resembling the benthic foraminifera Nonionella stella in the Oslofjord (Norway): morphological, molecular and paleo-ecological evidences

Specimens resembling the benthic foraminifera Nonionella stella (Cushman and Moyer, 1930), a morphospecies originally described from the San Pedro Basin, California, USA, were observed for the first time in the Oslofjord (Norway) in 2012. This study investigates the Oslofjord Nonionella population in order to confirm its nonindigenous species (NIS) status and assess its introduction time. Morphological characterisation based on SEM imaging complemented by molecular identification using small subunit (SSU) rDNA sequencing and assessment of the recent past record (sediment core), were performed on material collected in the Oslofjord in 2016. Examination of the dead fauna showed that specimens resembling N. stella only appeared recently in the Oslofjord, confirming the NIS status of this population. Moreover, DNA results indicate that the Oslofjord specimens differ genetically from N. stella sampled in the Santa Barbara Basin (California USA). Hence, we propose to use the name Nonionella sp. T1 for the specimens sampled in the Oslofjord for the time being. In the southern part of the Skagerrak, specimens morphologically similar to Nonionella sp. T1 were reported as NIS in the Gullmar fjord (Sweden) in 2011 and in the Skagerrak in 2015. Molecular data indicate that the two populations from Gullmar- and Oslofjords are identical, based on their SSU rDNA sequences. In addition, analyses of foraminiferal dead assemblages suggest that the population from the Gullmar fjord settled prior to the Oslofjord population, i.e. ~ 1985 and about 2010, respectively. This implies that Nonionella sp. T1 may have been transported from Sweden to Norway by northward coastal currents.

Molecular phylogenetic study of Bealius Massey & Hinds, 1970 (Nematoda: Hexatylina: Sphaerulariidae), with description of B. pinus n. sp. from Iran

Summary Bealius pinus n. sp. is described and illustrated based on its morphological, morphometric, and molecular characters. The new species is characterised by a body length of 1040 (920-1124) μm (females) and 985 (990-1043) μm (males), lip region continuous, 7.2 (7.0-8.0) μm broad, 3.3 (3.0-4.0) μm high, stylet length 8.4 (7.0-9.0) μm or ca 1.0-1.3 times the lip region diam., lateral fields with five or six incisures, the outer incisures crenate and inner incisures weakly crenate, and excretory pore situated 156 (141-170) μm from the anterior end. The basal pharyngeal bulb has a short posterior extension projecting dorsally into the intestine. Post-uterine sac absent. Functional males common in the population, spicules 29 (28-30) μm long. Tail cylindrical with broadly rounded terminus in the female and conical, arcuate, completely enveloped by a well-developed bursa in the male. The new species is compared with two known species of the genus, B. bisulcus and B. pissodi. Molecular phylogenetic studies of the new species using partial sequences of small subunit (SSU) rDNA revealed that it forms a clade with several nematode species belonging to genera in the Sphaerularioidea, namely: Sphaerularia, Paurodontella, Nothotylenchus, and Deladenus.

A new species of the genus Eudorylaimus Andrássy, 1959 (Nematoda: (Dorylaimida: Qudsianematidae) associated with Picea crassifolia in China.

A new species, Eudorylaimus piceae n. sp., extracted from rhizosphere soil of Picea crassifolia from Inner Mongolia, China was identified. The new species is characterized by these combined characters: body length of 1.03-1.27 mm; lip region distinctly offset; odontostyle 20-22 μm and 1.1-1.4 times the lip region diameter in length; odontophore 1.1-1.2 times the odontostyle length; basal expansion of pharynx occupying 42%-50.5% of the total neck length; genital system didelphic-amphidelphic; vulva transverse; vagina extending inwards 32%-43%; V value averaging more than 60; pars refringens vaginae with two trapezoidal sclerotizations and pars distalis vaginae weakly sclerotized; prerectum 72-107 μm long, 2.3-3.3 times the anal body diameter, and rectum 1-1.6 times the anal body diameter in length; tail conoid and bent ventrally, c' value 1.5-1.8 and males unknown. Phylogenetic analyses based on sequences of 18S small subunit rDNA and the D2-D3 expansion region of 28S rDNA are presented.

Morphology and molecular systematic of marine gregarines (Apicomplexa) from Southwestern Atlantic spionid polychaetes.

Gregarines are a common group of parasites that infect the intestines of marine invertebrates, and particularly polychaetes. Here, we describe for the first time four gregarine species that inhabit the intestines of three spionid species: Dipolydora cf. flava, Spio quadrisetosa and Boccardia proboscidea from the Patagonian coast, Argentina, using light and scanning electron microscopy and molecular phylogenetic analyses of small subunit (SSU) rDNA sequences. Even though the spionid species thrive in the same environments, our results showed a high host specificity of the gregarine species. Selenidium cf. axiferens and Polyrhabdina aff. polydorae were both identified from the intestine of D. cf. flava. The new species, Polyrhabdina madrynense sp. n. and Selenidium patagonica sp. n., were described from the intestines of S. quadrisetosa and the invasive species B. proboscidea, respectively. All specimens of D. cf. flava and S. quadrisetosa were infected by gregarines (P = 100%), recording the highest mean intensity values of infection (MI = 80; 60 respectively), in contrast to B. proboscidea (P = 60%; MI = 38). We associated this finding with the recent invasion of this host. It is expected that in the future, an increase of its population density might favour a rising intensity of this gregarine infection.

Integrative taxonomic approach of trypanosomes in the blood of rodents and soricids in Asian countries, with the description of three new species.

Trypanosoma lewisi (Kinetoplastea: Trypanosomatida: Trypanosomatidae) with a cosmopolitan distribution is the type species of the subgenus Herpetosoma, which includes ca. 50 nominal species isolated mainly from rodents. Since members of Herpetosoma in different host species have an almost identical morphology of bloodstream forms, these trypanosomes are referred to as 'T. lewisi-like', and the molecular genetic characterization of each species is necessary to verify their taxonomy. In the present study, we collected blood samples from 89 murid rodents of 15 species and 11 soricids of four species in Indonesia, Philippines, Vietnam, Taiwan, and mainland China for the detection of hemoprotozoan infection. T. lewisi and T. lewisi-like trypanosomes were found in the blood smears of 10 murid animals, which included Bandicota indica (two rats), Rattus argentiventer (one rat), and Rattus tiomanicus (two rats) in Indonesia; Rattus rattus (one rat) in the Philippines; and Niviventer confucianus (four rats) in mainland China. Furthermore, large- or medium-sized non-T. lewisi-like trypanosomes were detected in two soricids, Crocidura dracula in Vietnam and Anourosorex yamashinai in Taiwan, respectively. Molecular genetic characterization of the small subunit (SSU) ribosomal RNA gene (rDNA) and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene indicated that the trypanosomes from all the murid hosts had identical SSU rDNA or gGAPDH gene nucleotide sequences except for those in N. confucianus in mainland China. These N. confucianus-infecting trypanosomes also showed several unique morphological features such as smaller bodies, anteriorly positioned nuclei, and larger rod-shaped kinetoplasts when compared with T. lewisi trypomastigotes. Trypanosoma (Herpetosoma) niviventerae n. sp. is erected for this new species. Similarly, based on morphological and molecular genetic characterization, Trypanosoma sapaensis n. sp. and Trypanosoma anourosoricis n. sp. are proposed for the trypanosomes in C. dracula in Vietnam and A. yamashinai in Taiwan, respectively. More effort directed toward the morphological and molecular genetic characterization of the trypanosomes of rodents and soricids is required to fully understand the real biodiversity of their hemoflagellates.

Morphology, morphogenesis and molecular phylogeny of a new brackish water subspecies, Neourostylopsis flava paraflava nov. subsp. (Ciliophora, Hypotrichia, Urostylidae), with redefinition of the genus Neourostylopsis

The morphology, morphogenesis and small subunit (SSU) rRNA gene-based phylogeny of Neourostylopsis flava paraflava nov. subsp. were investigated. Neourostylopsis flava paraflava nov. subsp. was separated from N. flava flava nov. stat. by habitat (brackish water vs. fresh water), pretransverse cirri (absent in all specimens vs. present in 15% of specimens), the numbers of frontal cirri (8–15 vs. 6–8) and left marginal cirral rows (6–9 vs. 4–5). The main morphogenetic features of N. flava paraflava nov. subsp. are as follows: (1) streaks I–VI (or I–VIII, deduced from morphological data) produce the bicorona; (2) the oral primordium and frontoventral-transverse cirral anlagen in the opisthe are formed de novo on the cell surface; (3) the numerous macronuclear nodules fuse into a branch-like mass; (4) two pretransverse ventral cirri are formed initially but disappear in the later stages; and (5) some of the frontoventral-transverse cirral anlagen develop in a primary mode. Based on SSU rDNA sequence data, phylogenetic analyses show a close relationship between N. flava paraflava nov. subsp., N. flava flava nov. stat. and other Neourostylopsis species. An improved diagnosis for Neourostylopsis is provided: Urostylidae with five or more frontal cirri which form an indistinct or distinct bicorona; pretransverse cirri present or absent; transverse cirri present; buccal cirri present; two frontoterminal cirri; midventral complex composed of midventral pairs only; more than one row of marginal cirri on each side which derive from individual anlagen within each parental row; caudal cirri lacking; three dorsal kineties; endoral and paroral rather long, endoral straight, paroral distinctly curved anteriorly.

Morphology, morphogenesis, and phylogeny of an Anteholosticha intermedia (Ciliophora, Urostylida) population from the United States

A distinct population of Anteholosticha intermedia was isolated from soil in the Great Smoky Mountains of North Carolina, USA, and its morphology, morphogenesis and molecular phylogeny investigated by microscopic observations of live and protargol-prepared specimens and analyses of the sequence of small subunit (SSU) rDNA. Our population closely resembles the populations from Austria and Korea. Members of the genus Anteholosticha have been regarded as ontogenetically diverse, which is confirmed by the present work. The most noteworthy ontogenetic feature of the American population of A. intermedia is that the oral primordium in the proter appears apokinetally at the posterior end of the undulating membranes anlage at the beginning of division and then dedifferentiates midway through morphogenesis. Molecular phylogenetic analyses demonstrate, with high support, that the American population of A. intermedia is clearly distinct from congeners and branches as part of a sister lineage to the Bakuella–Urostyla clade that belongs to the major clade comprising the order Urostylida.

Revisiting the Parvilucifera infectans / P. sinerae (Alveolata, Perkinsozoa) species complex, two parasitoids of dinoflagellates

Members of the family Parviluciferaceae (Alveolata, Perkinsozoa) are the well-known dinoflagellate parasitoids along with Amoebophrya ceratii species complex and parasitic chytrid Dinomyces arenysensis and contain six species across three genera (i.e., Parvilucifera infectans, P. sinerae, P. rostrata, and P. corolla, Dinovorax pyriformis, and Snorkelia prorocentri) so far. Among Parvilucifera species, the two species, P. infectans and P. sinerae, are very similar or almost identical each other morphologically and genetically, thereby make it difficult to distinguish between the two. The only main difference between the two species known so far is the number of sporangium wall (i.e., 2 layers in P. infectans vs. 3 layers in P. sinerae). During sampling in Masan bay, Korea during the spring season of 2015, the dinoflagellate Akashiwo sanguinea cells infected by the parasite Parvilucifera were observed and this host-parasite system was established in culture. Using this culture, its morphological and ultrastructural features with special emphasis on the variation in the number of sporangium wall over developmental times, were investigated. In addition, the sequences of rDNA regions and β-tubulin genes were determined. The result clearly demonstrated that the trophocyte at 36 h was covered with 4 layers, and then outer layer of the sporocyte gradually degraded over time, resulting in wall structure consisting of two layers, with even processes being detached from 7-day-old sporangium with smooth surface, indicating that the difference in the number of layers seems not to be an appropriate ultrastructural character for distinguishing P. infectans and P. sinerae. While pairwise comparison of the large subunit rDNA sequences showed 100% identity among P. infectans / P. sinerae species complex, genetic differences were found in the small subunit (SSU) rDNA sequences but the differences were relatively small (11–13 nucleotides) compared with those (190–272 nucleotides) found among the rest of Parvilucifera species (P. rostrata and P. corolla). Those small differences in SSU rDNA sequences of P. infectans / P. sinerae species complex may reflect the variations within inter-strains of the same species from different geographical areas. Taken together, all morphological, ultrastructural, and molecular data from the present study suggest that they are the same species. Key words: inter-strain variation; parasitism; parasitoid; sporangium; synonymy

Molecular Phylogenetic Positions and Ultrastructure of Marine Gregarines (Apicomplexa) Cuspisella ishikariensis n. gen., n. sp. and Loxomorpha cf. harmothoe from Western Pacific scaleworms (Polynoidae).

Marine gregarines are unicellular parasites of invertebrates commonly found infecting the intestine and coelomic spaces of their hosts. Situated at the base of the apicomplexan tree, marine gregarines offer an opportunity to explore the earliest stages of apicomplexan evolution. Classification of marine gregarines is often based on the morphological traits of the conspicuous feeding stages (trophozoites) in combination with host affiliation and molecular phylogenetic data. Morphological characters of other life stages such as the spore are also used to inform taxonomy when such stages can be found. The reconstruction of gregarine evolutionary history is challenging, due to high levels of intraspecific variation of morphological characters combined with relatively few traits that are taxonomically unambiguous. The current study combined morphological data with a phylogenetic analysis of small subunit rDNA sequences to describe and establish a new genus and species (Cuspisella ishikariensis n. gen., n. sp.) of marine gregarine isolated from the intestine of a polynoid host (Lepidonotus helotypus) collected from Hokkaido, Japan. This new species possesses a set of unusual morphological traits including a spiked attachment apparatus and sits on a long branch on the molecular phylogeny. Furthermore, this study establishes a molecular phylogenetic position for Loxomorpha cf. harmothoe, a previously described marine gregarine, and reveals a new group of gregarines that infect polynoid hosts.

A red tide forming dinoflagellate Prorocentrum triestinum: identification, phylogeny and impacts on St Helena Bay, South Africa

:A non-toxic red tide with resulting anoxia was the cause of a major harmful algal bloom in St Helena Bay, South Africa, in February and March 2015. The red tide was observed along approximately 200 km of the Namaqua coastline extending well north of the Olifants River and southward into the bay. A maximum cell concentration of 14.32 × 106 cells l−1 was recorded in the southern reaches of the bay, and oxygen concentrations declined to as low as 0.06 ml l−1 in the shallow waters off Dwarskersbos. The hypoxia induced ecological fallout was significant with massive marine mortalities, including an estimated 415 tons of rock lobster (Jasus lalandii). The causative organism was identified morphologically, ultrastructurally and phylogenetically (using partial sequences of the 18S small subunit rDNA and the 28S large subunit rDNA) as Prorocentrum triestinum (Dinophyceae). Light and electron microscopy (both scanning and transmission) also facilitated identification of P. triestinum and addressed possible confusion with similar taxa. The global incidence of P. triestinum blooms is low, with hot spots limited to coastal areas in eastern China and the Mediterranean Sea, where blooms are sometimes associated with anthropogenic nutrient pollution. The harmful bloom documented here is the first exceptional bloom event with anoxic-related marine mortalities induced by P. triestinum in a major eastern boundary upwelling system such as the Benguela.

Genetic diversity and ecology of the planktonic foraminifers Globigerina bulloides , Turborotalita quinqueloba and Neogloboquadrina pachyderma off the Oman margin during the late SW Monsoon

The tropical waters of the Arabian Sea are among the richest biological areas of the world. The highly complex monsoonal system is particularly challenging for palaeoenvironmental study, which relies heavily upon understanding the modem-day ecology of planktonic foraminiferal assemblages and their geochemical signatures throughout the monsoonal cycle. Major upwelling responders such as G. bulloides, T. quinqueloba and N. pachyderma, typically associated with cooler mid to higher latitude ecosystems, are also found in number in the tropical Arabian Sea. Due to the more usual cooler water affinity of these morphospecies, the oceanographically isolated tropical upwelling ecosystem of the Arabian Sea potentially harbours new ecologically distinct genotypes (ecotypes). Samples were collected off the Oman margin at 15 stations towards the end of the summer monsoon to determine the genetic profiles of these morphospecies in both upwelling and open ocean regimes. Phylogenetic analysis of their small subunit (SSU) rDNA sequences revealed several new genetically distinct ecotypes. Two genetically divergent ecotypes of G. bulloides (Types Ia and IIf) were identified along the cruise track. Type Ia, a member of the G. bulloides warm water lineage, was found in both the upwelling and open ocean regions. The second genotype (IIf, a member of the G. bulloides cool water lineage, was found only in more marginal late upwelling cooler waters. Initial visual assessment of G. bulloides images suggests that it may be morphologically cryptic. Two highly divergent genotypes of T. quinqueloba (Types Ib and IIe) were also identified, which were largely confined to the eastern and northern Arabian Sea. Type He is a new member of the T. quinqueloba cool water lineage which points to its potential cool water affinity, but genotyping numbers are too low to confirm a specific association with upwelling. A new highly divergent genotype of N. pachyderma (Type VIII) was also identified at the western and southern stations. Comparison of global upwelling system genotype assemblages currently indicate little regional commonality. This complicates regional palaeoproxy understanding, since geochemical calibrations are known to be species and genotype specific. Detailed studies of the ecology and diversity of genotypes within each system should therefore be carried out to ensure the accuracy of palaeorecord interpretation.

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 (5-6μm 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.

Molecular Phylogenetic Positions of Two New Marine Gregarines (Apicomplexa)-Paralecudina anankea n. sp. and Lecudina caspera n. sp.-from the Intestine of Lumbrineris inflata (Polychaeta) Show Patterns of Co-evolution.

Gregarine apicomplexans are unicellular parasites commonly found in the intestines and coeloms of invertebrate hosts. Traits associated with the conspicuous feeding stage of gregarines, known as the trophozoite, have been used in combination with molecular phylogenetic data for species delimitation and the reconstruction of evolutionary history. Trophozoite morphology alone is often inadequate for inferring phylogenetic relationships and delimiting species due to frequent cases of high intraspecific variation combined with relatively low interspecific variation. The current study combined morphological data with small subunit (SSU) rDNA sequences to describe and establish two novel marine gregarine species isolated from the intestine of a polychaete host Lumbrineris inflata collected in British Columbia (Canada): Paralecudina anankea n. sp. and Lecudina caspera n. sp. The sister species to the host is Lumbrineris japonica, which can be found on the opposite side of the Pacific Ocean (Japan) and contains two different species of gregarine parasites: Paralecudina polymorpha and Lecudina longissima. Molecular phylogenetic analyses placed P.anankea n. sp. as the sister species to P.polymorpha and L.caspera n. sp. as the sister species to L.longissima. This phylogenetic pattern demonstrates a co-evolutionary history whereby speciation of the host (Lumbrineris) corresponds with simultaneous speciation of the two different lineages of intestinal gregarines (Paralecudina and Lecudina).

Molecular Characterization of Prototheca strains isolated in China revealed the first cases of protothecosis associated with Prototheca zopfii genotype 1.

In this study, six strains of Prototheca isolated in China from human patients diagnosed as protothecosis and cows with mastitis were characterized by polymerase chain reaction (PCR) and nucleotide sequencing of the ribosomal RNA gene (rDNA) and by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). The results indicated that three strains isolated from the human patients were P. zopfii genotype 1, revealing the first cases of human protothecosis associated with P. zopfii genotype 1. The remaining three strains were shown to be P. zopfii genotype 2. Interestingly, one strain isolated from the cerebrospinal fluid of the human patient appeared to have both of the genotype 1- and 2-specific alleles in the small subunit (SSU) rDNA although it was classified by MALDI-TOF MS as genotype 2. For genotyping of certain strains of P. zopfii, it may be necessary to comprehensively evaluate the diversity in the SSU rDNA sequences and the MALDI-TOF MS results.

Myxobolus chushi n. sp. (Myxozoa:Myxosporea) parasitizing Schizothorax niger (Heckel), a native cyprinid fish from Wullar Lake in Kashmir Himalayas

In the study, a new species, Myxobolus chushi n. sp. infecting gills of wild specimens of Schizothorax niger (Heckel) inhabiting Wullar Lake in Kashmir Himalayas, (J&K) India has been described based on morphology of the myxospore and using partial 18S rDNA sequencing. Pathological changes in the gills have been studied with the help of histological sections stained with Luna's method. Twenty fish specimens were examined, out of which four had oval, white plasmodia in gills measuring 2.0×0.5mm. The myxospores were spherical to ovoidal in shape with slightly attenuated posterior end, measuring 11.17±0.23 (10.60–11.40)μm in length and 9.14±0.06 (8.80–9.20)μm in width, having a prominent pore at the anterior end. The polar capsules were pyriform in shape, measuring 4.25±0.15 (4.00–4.40)μm in length and 2.38±0.27 (2.00–2.65)μm in width having polar filaments forming coils up to 5 in number. Parietal folds 9 in number present on the posterior part of the shell. The intensity of infection was recorded to be moderate as indicated by gill plasmodial index (GPI=2). The plasmodium was located in the vascular network occupying whole of the gill lamella therefore typed as intralamellar vascular type, LV3. Analysis of 18S small subunit (SSU) rDNA sequence of the isolate demonstrated 90% homogeneity with M. sp. KLT-2014 infecting scales of Labeo rohita from Myanmar and 89% with M. dermiscalis infecting scales of Labeo rohita from India.

Taxonomic revisions in the Microstromatales: two new yeast species, two new genera, and validation of Jaminaea and two Sympodiomycopsis species

Environmental sampling yielded two yeast species belonging to Microstromatales (Exobasidiomycetes, Ustilaginomycotina). The first species was collected from a leaf phylloplane infected by the rust fungus Coleosporium plumeriae, and represents a new species in the genus Jaminaea, for which the name Jaminaea rosea sp. nov. is proposed. The second species was isolated from air on 50% glucose media and is most similar to Microstroma phylloplanum. However, our phylogenetic analyses reveal that species currently placed in Microstroma are not monophyletic, and M. phylloplanum, M. juglandis and M. albiziae are not related to the type species of this genus, M. album. Thus, Pseudomicrostroma gen. nov. is proposed to accommodate the following species: P. glucosiphilum sp. nov., P. phylloplanum comb. nov. and P. juglandis comb. nov. We also propose Parajaminaea gen. nov. to accommodate P. albizii comb. nov. and P. phylloscopi sp. nov. based on phylogenetic analyses that show these are not congeneric with Jaminaea or Microstroma. In addition, we validate the genus Jaminaea, its respective species and two species of Sympodiomycopsis and provide a new combination, Microstroma bacarum comb. nov., for the anamorphic yeast Rhodotorula bacarum. Our results illustrate non-monophyly of Quambalariaceae and Microstromataceae as currently circumscribed. Taxonomy of Microstroma and the Microstromataceae is reviewed and discussed. Finally, analyses of all available small subunit rDNA sequences for Jaminaea species show that J. angkorensis is the only known species that possess a group I intron in this locus, once considered a potential feature indicating the basal placement of this genus in Microstromatales.