Ciliates in the Marsh Wetlands of Chongming Island: Taxonomy of Five Species and a Species List

To clarify the confusion surrounding species identification and state intraspecific variations, this research studied the morphological taxonomy of five species, Kentrophyllum cf. setigerum Petz et al., 1995; Zosterodasys shumerica Aliev, 1990; Frontonia tchibisovae Burkovsky, 1970; Neourostylopsis orientalis Chen et al., 2013; and Apoholosticha cf. sinica Fan et al., 2014, which were collected from Chongming Island's marsh wetlands. Kentrophyllum cf. setigerum possesses less somatic kineties than K. setigerum(19–26 right and 18–25 left kineties vs 35–40 right and 30 left kineties). Whether Z. shumerica is identical to Z. transversa Foissner et al., 1994, which was presented in a previous report, was negated. The improved diagnosis for the species is provided. Frontonia tchibisovae was confirmed to have three or four micronuclei, and the number of kinetosome rows in its peniculus 3 varied between China and Russia populations. The new population of N. orientalis bears cortical granules, which differs from the original population that has an arrangement of rows across the dorsal bristles. For A. cf. sinica, there are colorless cortical granules on the ventral side of the new population, whereas in the previously reported populations bright orange, spherical granules were clustered around dorsal cilia. Additionally, we sequenced the small-subunit(SSU) rRNA genes of K. cf. setigerum, N. orientalis, and A. cf. Sinica, and compared the new and reported populations. For the first time, we also provide a species list, including 33 ciliates collected from the wetlands based on living observation and silver staining.     

Cylindrotheca closterium Is a Species Complex as Was Evidenced by the Variations of rbcL Gene and SSU rDNA

The genus Cylindrotheca consists of a small group of marine diatoms with a few species described. Eleven isolates of diatoms identified as Cylindrotheca closterium morphologically were obtained from Jiaozhou Bay with their nuclear-encoded small-subunit ribosomal RNA （SSU rDNA） and chloroplast-encoded rbcL gene sequences determined in this study. Interestingly, very high sequence divergences of SSU rDNA and rbcL gene were found among these isolates, and numerous nucleotide variation of rbcL gene caused relatively few variation of deduced amino acid sequence. Phylogenetic analyses based on SSU rDNA and rbcL gene, respectively, grouped the isolates into 6 clades. Phylogenetic tree of SSU rDNA placed all the Cylindrotheca isolates together, separating them into two lineages clearly. LineageⅠ was composed of the eleven C. closterium isolates obtained in this study together with another C. closterium isolate, but some clades were not well supported. LineageⅠwas contained two C. closterium isolates and one C. fusiformis isolate. Phylogenetic analysis of rbcL gene also separated the Cylindrotheca isolates into two well-defined lineages. The eleven C. closterium isolates formed a lineage and all clades were supported strongly. Statistical comparisons of SSU rDNA indicated that the average distance within lineageⅠwas significantly higher than that of other microalgae species （P 〈0.01）. These results suggested the existence of cryptic species within C. closterium.     

Morphology and SSU rD NA Sequence Analysis of Two Hypotrichous Ciliates(Protozoa,Ciliophora, Hypotrichia) Including the New Species Metaurostylopsis parastruederkypkeae n.sp.

The morphology and phylogeny of two hypotrichous ciliates, Metaurostylopsis parastruederkypkeae n. sp. and Neourostylopsis flavicana(Wang et al., 2011) Chen et al., 2013 were investigated based on morphology, infraciliature and the small subunit(SSU) ribosomal RNA gene(r RNA) sequence. The new species, M. parastruederkypkeae n. sp. was identified according to its characteristics: body shape ellipsoidal, size about(165–200) ×(45–60) μm in vivo, cell color reddish; two types of cortical granules including wheat grain-like and yellow-greenish larger ones along the marginal cirri rows and dorsal kineties and dot-like and reddish smaller ones, grouped around marginal cirri on ventral side and arranged in short lines on dorsal side; 26–41 adoral membranelles; three frontal and one parabuccal, five to seven frontoterminal, one buccal, and three to six transverse cirri; seven to thirteen midventral pairs; five to nine unpaired ventral cirri, five to seven left and three to five right marginal rows; and three complete dorsal kineties. Phylogenetic analysis based on SSU r DNA sequences showed that both Metaurostylopsis and Neourostylopsis are monophyletic. As the internal relationship between and within both genera are not clear, further studies on the species in these two genera are necessary. The key characteristics of all known twelve Metaurostylopsis-Apourostylopsis-Neourostylopsis species complex were updated.     

Plant diversity and community structure of Brazilian Páramos

In Eastern South America, high altitude grasslands represent a mountain system that has a high number of endemic species. However, studies on the ecology of plant communities in these environments remain scarce. We aimed to evaluate the patterns of biodiversity and structure of plant communities from rocky outcrops in high altitude grasslands of three areas at the Caparaó National Park, southeastern Brazil, by sampling 300 randomly distributed plots. Then, we compared the floristic composition, relative abundance, and biological and vegetation spectra among areas. We classified species as endemic and non-endemic and verified the occurrence of endangered species. Species richness was evaluated by rarefaction analysis on the sampling units. The importance value and species abundance distribution(SAD) models were assessed. We also performed an indicator species analysis. We sampled 58 species belonging to 49 genera and 32 families. The number of species decreased with increasing altitude, with significant differences being observed among areas regarding richness, abundance, and cover. Of the total number of species, 10 are endemic to the Caparaó National Park and 17 are listed on the Brazilian Red List of endangered species. The dominant families on all peaks were Asteraceae and Poaceae. The SAD models showed lognormal and geometric distributions, corroborating the fact that 10 species that were common to all three areas were also the most dominant ones in the communities and showed the highest importance values, which ranged between 35% and 60%. Indicator species analysis revealed that 28 species(48.27%) were indicators. Of these, 42.85% had maximum specificity, meaning that they occurred only in one area. Thus, the number of species per life form ratio was similar among areas, yet vegetation spectra differed, especially for hemicryptophytes. The altimetric difference among the areas showed to be a very important driver in the community assembly, influencing the evaluated variables, however, other drivers as soil depth, slope and water could also influence the community structure on a smaller and local spatial scale.     

PSEUDOSMITTIA AIZAIENSIS, A NEW SPECIES OF ORTHOCLADIINAE (DIPTERA, CHIRONOMIDAE) FROM HUNAN PROVINCE OF CHINA

Male and female adult midges of Pseudosmittia aizaiensis sp. n. are described and illustrated with figures of morphological characters. The inferior volsella of gonocoxite with large lobes, the long but equal width gonostylus. and the small projection in basal part of outer margin are the characters that may separate this species from others. One larva of this genus is purely aquatic in habitat and is associated with Cricotopus bicinctus, Cardiociadius capucinns and Polypedilum sp. in the same algae debris (Spirogyra sp.) of the Dunghe River. Further, the larva's very short SI and SII bifids. its 5 pairs of lateral teeth on mentum and its absence of premandible brush are the characters that may also separate this species from any known species of this genus.     

Morphological observation of Epithemia ocellata (Ehr.) Kütz. (Bacillariophyceae) from China

The species of Epithemia ocellata (Ehr.) Kütz. from Northeastern China were observed by scanning electron microscopy (SEM). The studies on silica valve formation in different stages were made. At the early stage, a Y-shaped raphe sternum was found. Areolae were made from virgae and vimines. In immature valves, the canal formation before the raphe fissure is visible. The raphe system, virgae and vimines (areolae) might be the early siliceous elements of the valve during morphogenesis. The formations of virgae and vimines are similar to that of Gephyria media W. Arnott. Rows of silica papillae formed gradually on the face of the valve. After the areolae were covered with silica and form rows of papillae, the outer valve surface could become mature. Details of the internal costae were observed in mature valves.     

AreAcila divaricata and Acila mirabilis One Species or Two Distinct Species？ Evidence from COI Mitochondrial DNA

Acila divaricata （Hinds, 1843） and A. mirabilis （Adams and Reeve, 1850） are common benthic bivalves in China. A number of researchers have proposed that the latter species is a junior synonym of the former species. Because of morphological similarities, it is difficult to distinguish these two species based on visual examination only. For better understanding of their taxon-omy, the mitochondrial CO1 gene fragments of five individuals of A. divaricata from the East China Sea and six individuals of A. mirabilis from the Yellow Sea were sequenced in this study. The phylogenetic relationships of the obtained COI sequences, together with nineteen sequences of three species of the genus Nucula, were analyzed. The pairwise intra- and inter-specific distances for the CO1 sequences ranged from 0.002 to 0.017 and from 0.128 to 0.134, respectively, and no overlap was found. Phylogenetically, A. divaricata and A. mirabilis form distinct clades and cluster into a sister to all other Nucula species. The results indicated that A. di-varicata and A. mirabilis are two distinct species. The differences in the morphology and distribution between the two species were briefly discussed.     

Taxonomy,Morphology and Phylogeny of a New Oligotrich Ciliate-Omegastrombidium hongkongense n. sp.(Protozoa: Ciliophora) from Clear Water Bay,Hong Kong

One new marine oligotrich ciliate, Omegastrombidium hongkongense n. sp., was isolated from a bloom of Noctiluca scuntillans near Port Shelter, Hong Kong. The morphology and infraciliature of this new species were studied on both living and protargol-stained specimens. Its phylogenetic position was discussed based on the sequence of the small subunit r RNA gene. O. hongkongense is different from its congeners with special characters. The cells are usually heart-shaped, and the cell size usually is(20–35) ×(20–30) μm in vivo. Its deep buccal cavity extends obliquely to about 1/2 of cell length. It shows prominent apical protrusion. The adoral zone of membranelles is divided into 17–19 collar membranelles and four buccal membranelles. It has one ball-like macronucleus. The girdle kinety forms a closed loop which obliquely surrounds the body. The ventral kinety and thigmotactic membranelles are not observed. The SSU r RNA sequence of O. hongkongense was close to those of Strombidium paracalkinsi and Varistrombidium kelum with approximately 99% similarity. In the phylogenetic trees, O. hongkongense can be grouped with O. elegans and V. kielum species with very low support(16% ML).     

A New Record of Ponyfish Deveximentum megalolepis(Perciformes: Leiognathidae) in Beibu Gulf of China

A new record ponyfish, Deveximentum megalolepis Mochizuki and Hayashi, 1989, was documented based on its morphological characteristics and DNA barcode. Fifty specimens were collected from Beibu Gulf of China and identified as D. megalolepis by morphological characterization. The coloration, meristic traits, and morphometric measurements were consistent with previously published records. In general, it is a silver-white, laterally compressed and deep bodied ponyfish with 6–9 rows of scales on cheek; scale rows above lateral line 6–8; scale rows below lateral line 14–17. Mitochondrial cytochrome oxidase I subunit(COI) gene fragment was sequenced for phylogenetic analysis. There is no sequence variation of COI gene between the specimens collected in this study. The genetic distances between D. megalolepis and other congeneric species range from 3.6% to 14.0%, which were greater than the threshold for fish species delimitation. The COI sequence analysis also supported the validity of D. megalolepis at genetic level. However, the genetic distance between Chinese and Philippine individuals was about 1.2% and they formed two lineages in gene tree, which may be caused by the geographical distance.     

Two new species of the genus Cobitis Linnaeus(Teleostei:Cobitidae) from southern China

Two new species of the genus Cobitis from southern China,C.hereromacula from the Luohe River in Guangdong Province and C.baishagensis from the Nandujiang River in Hainan Province,are described and illustrated here.C.hereromacula can be distinguished from its congeners by possessing the following combination of characteristics;absence of the second and third pigmentary zones of Gambetta;13–16 oval blotches on the dorsum and 10–13 vertical,elongated triangular blotches below the midlateral line with more than 20 vertical dark brown bars between them;6–7 narrow rows of dark spots on the caudal fin;a vertical oval spot smaller than the eye diameter on the upper part of the caudal peduncle;pointed mental lobes of the lower lip pointed with a slightly filiform tip;one slender and long needle-shaped lamina circularis at the base of the first branched ray of the male pectoral fins.C.baishagensis can be distinguished from its congeners by the fourth Gambetta zone being covered by 10–12 transverse elongated blotches;4–5 narrow rows of dark spots on the caudal fin;a vertical blotch smaller than the eye diameter on the upper part of the caudal peduncle;males with a slender and long needle-shaped lamina circularis at the second branched pectoral fin ray in males;large scales with a slightly large focal zone;undeveloped mental lobes with a lower lip that does not end posteriorly in a filiform tip.     

Molecular identification of Prorocentrum （Dinophyceae） species

A fragment of a large sub-unit ribosomal DNA (LrDNA) of 12 strains of Prorocentrum species was amplified by polymerase chain reaction (PCR). The PCR products were digested by 3 restriction endonucleases (Cfo Ⅰ, Hae Ⅲ, and RSA Ⅰ) and then resolved in agarose gels. Results show that different species had different RFLP patterns, except for P arcuatum (ME 131), which had the same pattern to P. micans (ME 160 and 04).The same fragment of 19 strains of the genus was also amplified and subjected to denaturing gradient gel electrophoresis (DGGE). 11 different patterns were resolved. Different cultures of a same species had the same pattern. The results of RFLP and DGGE analyses showed that eight newly isolated epibenthic Prorocentrum species were different from each other, and also from other cultured ones examined in this study. P arcuatum(MEI32) could not be differentiated from P. micans (MEI60 and 04), it was probably mis-identified, since they are quite different morphologically. P. redfieldii (MEI38) could also not be distinguished form P. triestinium(MEI32), it should be regarded as a synonym ofP. triestinium. Unexpectedly, a restriction site was found in P.micans, compared with previous sequence data.     

How do altitude and soil properties influence the taxonomic and phylogenetic structure and diversity of Brazilian páramo vegetation?

Altitude and environmental variables such as edaphic properties are considered determinants of species distribution and community composition in mountain ecosystems.Here,we aimed to outline the effects of distinct mountain peaks,altitude and soil properties on community composition,species density,phylogenetic structure and diversity of angiosperm páramo communities from the Serra do Brigadeiro State Park,Minas Gerais,southeastern Brazil.For that,we identified all angiosperm species found in 300 plots(1 m×1 m)from three mountain peaks,measured soil depth and analyzed soil fertility and texture in each plot.To reduce the number of soil variables and species composition,we computed principal coordinates based on soil properties and principal coordinates based on species-plot matrix for each plot.Furthermore,we computed the standard effect sizes of the mean phylogenetic pairwise distance and the mean nearest phylogenetic taxon distance for each plot to investigate differences in the degree of relatedness among coexisting species.We compared differences in response variables between peaks and modelled them in function of altitude and principle components of soil properties using mixed effect models.Species density and phylogenetic diversity differed between peaks,but,contrary to the previous findings,no relationships between species richness or phylogenetic diversity and altitude or soil properties were found,indicating that further investigations are necessary to understand the altitude-biodiversity relationship in Brazilian páramo vegetation.Community composition differed between peaks and depended on altitude,soil properties and interactions between them,indicating that upward shifting of bioclimatic conditions due to climate changes may alter communities of this ecosystem.Phylogenetic structure differed between peaks and was influenced by altitude and soil properties.As phylogenetic clustering increased with altitude,eventual upward movements of species in Brazilian páramo vegetation due to climate change may alter community composition and the degree of relatedness among coexisting species,increasing the risk of species from higher altitudes to disappear.Therefore,conservation priorities arise for higher landscape portions,where these high altitude species may find refuges.     

Phylogenetic analysis of cultivable bacteria isolated from Arctic sea-ice

Phylogenetic analysis based on 16S rDNA of 8 strains of cultivable bacteria isolated from Arctic sea-ice was studied.The results showed that strain BJ1 belonged to genus Planococcus,which was a genus of low mole percent G C gram-positive bacteria;strain BJ6 belonged to genus Burkholderia of β-proteobacteria and the rest 6 strain all belonged to γ-proteobacteria,of which strain BJ8 was a species of Pseudoalteromonas,strain BJ2-BJ5 and BJ7 were members of genus Psychrobacter.Phylogenetic analysis also indicated that bacteria of genus Psychrobacter of the isolates formed a relatively independent phylogenetic cluster in comparison with other bacteria belonged to genus Psychrobacter.     

A NEW SPECIES OF CILIATES, HEMIOPHRYS POLYMICRONUCLEI SP. NOV. FROM DONGHU LAKE, HUBEI PROVINCE

A new species of ciliates Hemophrys polymicronuclei sp. nov. from Donghu Lake is described. It is characterized by its colorless to yellowish, soft, flat and spindle-shaped body with four micronuclei.The right side of the body is a little more convex than the left side, and more transparent at both anterior and posterior ends. Its slit-like cytostome accounts for 1/4 of the body length on the left side without any trichocyst. There are two rows of trichites near the right anterior end ,6 - 8 straight bristles on the pellicle opposite the cytostome, 23-25 rows of kinetides on the right side of the body, two ellipsoid macronuclei and many contractile vacuoles in the cytoplasm.     

Species identification and phylogenetic analysis of genus Nassarius(Nassariidae)based on mitochondrial genes

Genus Nassarius contains many subgenera, such as Zeuxis, Telasco, Niotha, Varicinassa, Plicarcularia, Nassarius s. str. and Reticunassa. On the basis of morphological characteristics of the shell and radula and sequences of mitochondrial cytochrome oxidase subunit I (COI) and 16S rRNA genes, Nassarius specimens collected from the South China Sea were identified and phylogenetically analyzed. Although Nassarius sp. and Nassarius (Varicinassa) variciferus were morphologically different in their shells, few variations were found among their radular teeth and sequences of mtCOI and mt16S RNA genes. Therefore, Nassarius sp. should be classified as N. (Varicinassa) variciferus. Nassarius (Zeuxis) sp. has only a subtle difference from Nassarius (Zeuxis) algidus on the shell, but it shows obvious differences in radular teeth and DNA sequence, indicating that they are two distinct species. Sequence divergence of mtCOI and mt16S RNA genes within Nassarius species was much lower than that between species, suggesting that these two genes are suitable for Nassarius species identification. Phylogenetic analysis (neighbor-joining and maximum parsimony) based on mtCOI and mt16S rRNA genes revealed the presence of two groups in genus Nassarius and a closest relationship between subgenera Zeuxis and Telasco. Species of subgenus Plicarcularia did not form a single clade. The molecular phylogeny was not congruent with the previous morphological phylogeny. The subgeneric divisions of genus Nassarius appear to be uncertain and unreliable.     

Comparative Analysis of Mitochondrial Control Region Sequence from Three Flatfish Species （Pleuronectidae）

The 5‘-end of the mitochondrial control region sequences of three flatfishes (Pleuronectiformes: Pleuronectidae) were amplified and sequenced. These sequences were compared with those of other three Pleuronectids species retrieved from GenBank. A phylogenetic tree was constructed based on the partial control region sequences. The results of phylogenetic analysis are consistent with those of conventional systematics. Compared to previous studies, the structure of the 5‘-end of mitochondrial control region was analyzed. The terminal associated sequence motif and its complementary motif were identified at the 5‘-end of the sequences. A conserved sequence block, named as CM5‘ d, was identified in the 5‘-end of control region sequences in all Pleuronectids. Another central conserved sequence block, named as CSB-F, was detected in the central conserved blocks.     

REPRODUCTION ECOLOGY OF FISHES IN JIAOZHOU BAY WATERS

Observations on gonad development of 84 species of marine fishes in Jiaozhou Bay waters showed the breeding periods of more than 50 of these species are mostly from April to August, with peaks between May to June. A great many of the species spawn in the warm season, only a few species in the cold season. The number of spawning species increases with the rise and decreases with the fall in water temperature. The absolute fecundity of 50 teleost species, which usually increases with body length and weight, was estimated to range from about 300 to 9,000,000 eggs. The reproductive patterns of fishes in Jiaozhou Bay waters are discussed.     

STUDY ON THE PHYTOPLANKTON.IN A LARGE RESERVOIR

A comprehensive study on the community structure and function of the phytoplankton in Taipinghu Reservoir, the largest reservoir (9400 ha) in Anhui Province, China, was carried out dur ing 1985- 1986. A total of 175 species of algae belonging to 8 phyla and 87 genera was noted. The composition of phytoplankton was dominated by species of Chlorophyta, Cyanophyta and Bacillariophyta. The species number, cell density and biomass of the three groups were respectively 88%, 86.5% and 78.9% of the total phytopiankton. The weighted annual average biomass was 1.52 mg/L and cell density was 1.43 x 106 ind/L. The growth maximum was observed in summer. Diatoms were abundant in the region adjoining a river. A large number of flagellated algae such as Euglena and Chromulina occurred in the artificial fish culture bay.It can be inferred from the algal composition and total nitrogen concentration (1.48mg/L) that this reservoir is a mesotrophic water body that had undergone slight natural eutrophication.Presented are an     

A Preliminary Phylogenetic Analysis ofLuidia （Paxillosida： Luidiidae） from Chinese Waters with Cytochrome Oxidase Subunit I （COI） Sequences

Luidia Forbes （Paxillosida： Luidiidae） are common soft bottom sea stars with 49 described species. Because of substan- tial morphological diversity, the taxonomy of the genus is complex and hasn＇t been resolved definitely. In order to resolve general taxonomic issues, and determine species boundaries and phylogenetic relationships within the genus Luidia, the sequences of tyro- chrome oxidase subunit I （COI） gene from 24 specimens of Luidia, belonging to eight taxa in Chinese waters, were studied. Three sequences of two species in genus Luidia from GenBank were used to analyze the phylogenetie relationships. The molecular phy- logeny exhibited three main clades, each with strong bootstrap support： Clade A including Luidia quinaria from the Sea of Japan; Clade B including seven nominal species （L. quinaria von Martens, L. yesoensis Goto, L. ehangi Liu, Liao and Li, L. orientalis Fisher, L. avicularia Fisher, L. longispina Sladen and L. hardwicki Gray） from Chinese waters; and Clade C including L. maculata Miiller ＆ Troschel from Chinese waters. Our molecular phylogeny results support the morphological Quinaria-Group and Alter- nata-Group assigned by D6derlein. Seven nominal species we sampled do not exhibit genetic distances that are large enough to rec- ognize them as separate species. Cryptic species may exist in ＇Luidia quinaria＇ from the Yellow Sea and the Sea of Japan. Meaning- ful morphological characters need further investigation in Luidia.