Species of Gonostomum and Paragonostomum (Ciliophora, Hypotrichida, Oxytrichidae) from the Valley of Flowers, India, with descriptions of Gonostomum singhii sp nov, Paragonostomum ghangriai sp nov and Paragonostomum minuta sp nov

Soil samples taken from the Valley of Flowers, a component of the Nanda Devi Biosphere Reserve in the Himalayan regions of India showed the presence of twenty two free living species of ciliates. There is a preponderance of species which exhibit oral ciliature and ontogenesis in the Gonostomum pattern. Of the four species of the genus Gonostomum, three viz., G affine, G gonostomoida and G kuehnelti are similar to described species; Gonostomum singhii is new. The two species of genus Paragonostomum viz., P minuta and P ghangriai are new. The three new species are described in the present paper. All these species show prominent hypertrophied ciliary structures. Their paroral membranes reveal characteristic differences with respect to their position, number of constituent cilia and the distance between adjacent cilia. It is proposed that such species specific features of the paroral membrane have a bearing in exercising different food organism preferences as they co-exist at many sites. This single factor has possibly played an important role in species diversification of this group of hypotrichs in this isolated habitat.     

Evaluation of potential health risks to Eastern Elliptio (Elliptio complanata) (Mollusca: Bivalvia: Unionida: Unionidae) and implications for sympatric endangered freshwater mussel species

Conservation efforts for freshwater musselspecies require identification and evaluationof potential health risks to populations. Sampling large numbers of individual mussels,however, could damage the small, fragile,extant populations of imperiled mussel species. In this study, a small sample size was designedto reduce lethal sampling, costs, andenvironmental disruption, while still allowing95% confidence in detecting health risks of25% or greater prevalence in the population. Health assessments were conducted on twentyspecimens of the Eastern Elliptio, Elliptio complanata, collected from two NorthCarolina sites as part of a survey to evaluatepotential disease threats to mussels in theregion. Bacteriological sampling of thegastrointestinal tracts yielded 18 aerobicbacterial species, of which Aeromonashydrophila (55.0%), Enterobacter spp.(40.0%), and Bacillus spp.(30.0%) were predominant. Histologicallesions of internal organs included mild tomoderate digestive gland atrophy andinflammation in one mussel, and mild tomoderate parasitism in several individuals. Adistinct difference in parasite prevalence wasevident between infections in E.complanata from the two collection sites. Thetrematode metacercaria of Homalometronarmatum and what appears to be three gillciliate species, the most abundant beingtentatively identified as the scyphidiidperitrich Mantoscyphidia sp., were foundin mussels from one site only. This studydemonstrates a comprehensive diagnosticapproach incorporating multiple modalities toassess the health status of mussel populations,while minimizing the sample size required toobtain valuable information. Furthermore, thisstudy provides baseline health data of E.complanata at two sites in south-central NorthCarolina and suggests the potential usefulnessof E. complanata as an environmentalbioindicator of health risks to sympatricthreatened freshwater mussel populations.     

四种淡水缘毛类纤毛虫的形态学研究

通过活体显微观察及银染法对采自武汉东湖的4种固着缘毛类纤毛虫: 粗茎睫纤虫Ophrydium crassicaule Penard, 1922, 念珠伪钟虫Pseudovorticella monilata (Tatem, 1870) Foissner & Schiffmann, 1974, 垂盖虫Opercularia nutans (Ehrenberg, 1831) Stein, 1854和一个鞘居虫未定种Vaginicola sp.的活体、纤毛图式、银线系以及细胞核的形态学特征进行详细的描述, 补充和完善了缘毛类纤毛虫的形态学资料, 为纤毛虫分类和系统发育学的研究提供重要信息。此外, 首次在伪钟虫中观察到有性生殖现象, 为纤毛虫生殖进化的研究提供了基础资料。     

Description and Molecular Phylogeny of a Novel Hypotrich Ciliate from the Soil of Marche Region,Italy; Including Notes on the MOSYSS Project

The morphology and morphogenesis during cell division of a new stylonychine hypotrich, Rigidocortex quadrinucleatus n. sp., were investigated using live observation and protargol staining. The new species was isolated from soil samples collected from an organic farm in the Marche Region, Italy, in framework of the MOSYSS project. Rigidocortex quadrinucleatus is characterized as follows: cell size about 180 × 80 μm in vivo; four ellipsoidal macronuclear nodules; 44 adoral membranelles: 18 fronto‐ventral‐transverse cirri consisting of three frontal, four frontoventral, one buccal, three ventral, two pretransverse, and five transverse cirri; dorsal kinety 3 with multiple fragmentation; resting cyst with hyaline ridges. Rigidocortex quadrinucleatus mainly differs from the type species R. octonucleatus in having four (vs. eight) macronuclear nodules. Rigidocortex quadrinucleatus can be easily confused with Sterkiella cavicola since both have a rather similar ventral ciliature; however, they can be separated by the slightly higher number of cirri in the left marginal row that runs along the posterior cell's margin in R. quadrinucleatus. Morphogenesis on the ventral surface is highly similar to that of Sterkiella species, but differs significantly on the dorsal surface (multiple vs. simple fragmentation of dorsal kinety 3). Phylogenetic analyses based on SSU rRNA gene sequences consistently place the new species within the stylonychine oxytrichids, clustering closer to Gastrostyla steinii than to S. cavicola.     

Signalling in ciliates: long‐ and short‐range signals and molecular determinants for cellular dynamics

In ciliates, unicellular representatives of the bikont branch of evolution, inter‐ and intracellular signalling pathways have been analysed mainly in Paramecium tetraurelia, Paramecium multimicronucleatum and Tetrahymena thermophila and in part also in Euplotes raikovi. Electrophysiology of ciliary activity in Paramecium spp. is a most successful example. Established signalling mechanisms include plasmalemmal ion channels, recently established intracellular Ca²⁺‐release channels, as well as signalling by cyclic nucleotides and Ca²⁺. Ca²⁺‐binding proteins (calmodulin, centrin) and Ca²⁺‐activated enzymes (kinases, phosphatases) are involved. Many organelles are endowed with specific molecules cooperating in signalling for intracellular transport and targeted delivery. Among them are recently specified soluble N‐ethylmaleimide‐sensitive factor attachment protein receptors (SNAREs), monomeric GTPases, H⁺‐ATPase/pump, actin, etc. Little specification is available for some key signal transducers including mechanosensitive Ca²⁺‐channels, exocyst complexes and Ca²⁺‐sensor proteins for vesicle–vesicle/membrane interactions. The existence of heterotrimeric G‐proteins and of G‐protein‐coupled receptors is still under considerable debate. Serine/threonine kinases dominate by far over tyrosine kinases (some predicted by phosphoproteomic analyses). Besides short‐range signalling, long‐range signalling also exists, e.g. as firmly installed microtubular transport rails within epigenetically determined patterns, thus facilitating targeted vesicle delivery. By envisaging widely different phenomena of signalling and subcellular dynamics, it will be shown (i) that important pathways of signalling and cellular dynamics are established already in ciliates, (ii) that some mechanisms diverge from higher eukaryotes and (iii) that considerable uncertainties still exist about some essential aspects of signalling.     

四膜虫eRF1识别终止密码子的功能结构域

原生动物的一些纤毛虫中终止密码子发生重分配现象,将1个或2个终止密码子翻译为氨基酸.目前对这一现象的发生机制仍无合理解释．近年来,对蛋白质合成终止过程中肽链释放因子(eukaryotic polypeptide release factor, eRF)结构和功能的深入研究,为揭示终止密码子的重分配机制提供了重要的线索．本实验以具有终止密码子识别特异性的四膜虫Tt-eRF1为研究对象,将其中与密码子识别有关的GTx、NIKS和Y-C-F关键模体(motif) 引入识别通用终止密码子的酵母Sc=eRF1中,构建成各种嵌合体eRF1．利用双荧光素酶报告系统和细胞活性实验,分析关键模体及其周边的氨基酸对eRF1识别终止密码子性质的影响．结果表明,GTx和NIKS模体一定程度上决定eRF1识别终止密码子第1位碱基U和第2位碱基A;Y-C-F模体决定eRF1识别终止密码子UGA的第2位碱基G．模体内及其相邻氨基酸定点突变分析进一步支持以上结果．本研究推测,eRF1在进化过程中一些关键模体结构的改变决定其识别终止密码子的特异性,只能识别3个终止密码子中的1个或2个．随后,由于tRNA基因的突变产生阻抑性tRNA,促成终止密码子在原生动物纤毛虫中的重新分配．     

Increase in photosynthetic efficiency as a strategy of planktonic organisms exploiting deep lake layers

1. The photosynthetic efficiencies of the mixotrophic ciliate Ophrydium naumanni and the autotrophic dinoflagellate Gymnodinium paradoxum were investigated using laboratory and field experiments in Lake Moreno Oeste (41°5′S and 71°33′W, 758 m a.s.l.), in the Nahuel Huapi System (North Patagonia, Argentina). 2. The effect of different underwater light intensities on net primary production (NPP) was assessed during one summer. Additionally, laboratory experiments were carried out to obtain photosynthesis‐irradiance response curves for each species. 3. Ophrydium naumanni and G. paradoxum dominated the metalimnetic (30 m depth) deep chlorophyll maximum (DCM) in the lake. 4. Despite these deep higher abundances, the cell‐specific production of both species was higher at 10 m than at 30 m (DCM) depth. In addition, at 5 m depth, NPP was reduced by PAR + UV‐A radiation. 5. Both species exhibited a positive NPP at very low irradiance but the mixotrophic ciliate was more efficient in exploiting the DCM irradiance level both in situ and at comparable light intensities in laboratory experiments. Light acclimatised O. naumanni showed a higher NPP at lower irradiances and photoinhibition at medium and high irradiances. 6. Under the strong wind‐driven turbulence commonly found in Patagonian lakes, organisms cannot select their position in the epilimnetic water column and will be dragged to potentially harmful UV radiation levels. Thus, metalimnetic DCM colonisation by these two species represents a tradeoff between higher survival and lower cell‐specific NPP.     

大型纤毛虫总RNA的提取方法

以冠突伪尾柱虫为材料,建立了一种适合于富含RNase和多糖的大型纤毛虫总RNA的提取方法。该方法采用异硫氰酸弧和β-巯基乙醇联合快速变性,酚、氯仿和异戊醇抽提,同时在变性剂存在的条件下两次选择性地沉淀RNA,从而有效地去除了DNA、蛋白质和多糖。所得RNA样品经电泳、紫外分光光度法检测和RT-PCR分析,证实为完整、均一且无基因组污染的总RNA.这为构建冠突伪尾柱虫有小核系与无小核系之间的削减文库、筛选两细胞系差异表达的基因奠定了基础。     

Mixotrophic cryptophytes and their predators in the Dry Valley lakes of Antarctica

1. The ingestion rates of planktonic, mixotrophic cryptophytes in two perennially ice-covered Antarctic lakes in the McMurdo Dry Valleys, were investigated during the summer of 1997–1998.
2. In Lake Fryxell, which is meromictic, ingestion rates increased with depth in November and were highest in a cryptophyte maximum close to the chemocline. In Lake Hoare, which is unstratified and freshwater, there was no significant difference in ingestion rates with depth. In both lakes, the highest ingestion rates occurred in early summer, decreasing in December and January. Ingestion rates varied between 0.2 bacteria cell⁻¹ h⁻¹ and 3.6 bacteria cell⁻¹ h⁻¹.
3. During November, mixotrophic cryptophytes removed up to 13% of bacterial biomass day⁻¹ and had a greater grazing impact than heterotrophic nanoflagellates (HNAN). As summer progressed, the grazing impact of cryptophytes and HNAN became similar.
4. The maximum depth of cryptophytes in Lake Fryxell was predated by a population of the ciliate Plagiocampa. Plagiocampa had an ingestion rate of 0.13–0.19 cryptophytes cell⁻¹ h⁻¹. The grazing impact on the cryptophyte community was insignificant. However, the ciliate appeared to be indulging in temporary mixotrophy, sequestering the cryptophytes for a number of weeks before digesting them.
5. It is suggested that mixotrophy is an important survival strategy in the extreme lake ecosystems of the McMurdo Dry Valleys.