Macronuclear chromatin of Blepharisma japonicum compared to that of Tetrahymena pyriformis

Abstract The macronuclear chromatin of the ciliate Blepharisma japonicum was studied by electrophoretic and immunological techniques as well as by micrococcal nuclease digestion and circular dichroism spectroscopy. Under these experimental conditions the macronuclear chromatin of B. japonicum was compared to that of Tetrahymena pyriformis . Data obtained through this analysis showed that the macronuclear chromatin of B. japonicum is structurally more relaxed than that of T. pyriformis . A perchloric acid soluble polypeptide, referred to as P3, is the only polypeptide of B. japonicum chromatin to appear immunologically related to the H1 histone of T. pyriformis . It is suggested that this P3 polypeptide in B. japonicum should be considered functionally equivalent to the T. pyriformis H1 histone, even though it differed from it both in terms of molecular mass and relative concentration.     

Evidence for natural transformation of Bacillus subtilis in foodstuffs

The effect of foodstuffs on the natural transformation of Bacillus subtilis was investigated. As examples of complex food matrices milk with various fat contents as well as chocolate milk were used. The frequencies of transformation varied with the fat content and ranged between 3.8×10⁻⁴ and 1.4×10⁻³. Highest frequencies of about 3×10⁻³ were observed in chocolate milk with 1.5% fat. Development of competence was observed in chocolate milk, resulting in maximal transformation frequencies upon incubation for 10–12 h at 37°C.     

Tetrahymena australis (Protozoa,Ciliophora): A Well‐Known But “Non‐Existing” Taxon – Consideration of Its Identification,Definition and Systematic Position

A cryptic species of the Tetrahymena pyriformis complex, Tetrahymena australis, has been known for a long time but never properly diagnosed based on taxonomic methods. The species name is thus invalid according to the International Code of Zoological Nomenclature. Recently, a population isolated from a freshwater lake in Wuhan, China was investigated using live observations, silver staining methods and gene sequence data. This organism can be separated from other described species of the T. pyriformis complex by its relatively small body size, the number of somatic kineties and differences in sequences of two genes, namely the small subunit ribosomal RNA (SSU rRNA) and the mitochondrial cytochrome c oxidase subunit I (cox1). We compared the SSU rRNA gene sequences of all available Tetrahymena species to reveal the nucleotide differences within this genus. The sequence of the Wuhan population is identical to two sequences of a previously isolated strain of T. australis (ATCC #30831). Phylogenetic analyses indicate that these three sequences (X56167, M98015, KT334373) cluster with Tetrahymena shanghaiensis (EF070256) in a polytomy. However, sequence divergence of the cox1 gene between the Wuhan population and another strain of T. australis (ATCC #30271) is 1.4%, suggesting that these may represent different subspecies.     

Resting Cysts of the Pigmented Ciliate Blepharisma sinuosum Sawaya, 1940 (Ciliophora: Heterotrichea)

The morphology of Blepharisma sinuosum resting cysts and the dynamics of pigmentation at different stages of encystment are presented for the first time. Cyst morphometrics are similar to other Blepharisma species, with three‐wall layers, bacteria surrounding the ectocyst, a conical plug, and wrinkly surface toward the plug in mature stages. The vegetative moniliform macronucleus changes to a horseshoe shape, and at early stages, the cystic cytoplasm is homogeneously pigmented, comprising a contractile vacuole; later, pigments polarize toward the plug, decorate the cortical layer, and become brownish. This work reinforces the potential role of pigment dynamics on cyst biology.     

中国土壤原生动物新纪录种(纤毛虫门：多膜纲；异毛目)

报道了首次在中国土壤中鉴定到的、隶属于纤毛虫门多膜纲异毛目的７种原生动物。这些种类是泥炭藓苔扭虫、透明赭虫、长赭虫、透明扭头虫、弯曲扭头虫、旋短端突虫和梅氏扁豆虫。     

Description of Opisthonecta Matiensis N. Sp. (Protozoa, Ciliophora), A New Peritrich Ciliate From Wastewater

Opisthonecta matiensis n. sp. was isolated from the inlet water of a wastewater treatment plant near Madrid, Spain, and studied in vivo, with silver methods, and using electronic and indirect immunofluorescence microscopy. This new species shows an amphora-like cell shape and has a size of 45-73 microns (x 58.2) x 25-40 microns (x 31.3). The oral infraciliature is formed by one haplokinety, three polykineties, and a short row of kinetosomes (epistomial membrane). The aboral infraciliature is made up of the trochal band and the scopula. From the trochal band arise three fibrillar systems: oral fibers, aboral fibers, and oblique fibers. The myoneme system is composed of a delicate peristomial ring, longitudinal branched fibers that reach the trochal band and of radial fibers extending from the scopula to the trochal band. The silverline system consists of an average of 147 lines. This new species is separated from other known forms by its smaller size, the presence of one single vacuole, and its higher number of silverlines.     

A New Species of the Genus Metacystis (Ciliophora,Prostomatida, Metacystidae) from a Wastewater Treatment Plant

ABSTRACT. Unusual prostomatid specimens were found in the biological reactor of a wastewater treatment plant in a health resort in Valencia, Spain. These ciliates were attached to flocs unlike other free‐swimming prostomatid ciliates described to date in the mixed liquor of activated sludge plants. The morphological study of this species led to a typically different combination of characteristics: elongated cell shape, 20–30 somatic kineties, 2 perioral kineties, and 1 circumoral kinety, 1 large vacuole protruding at the terminal end, a lorica tapered toward the aperture with a smooth neck, and 11–16 annular ridges. These characteristics place this representative as a new species of the genus Metacystis—Metacystis galiani n. sp. This species became the dominant population within the biological reactor when high values of conductivity (4,244 mS/cm) and temperature (26.8 °C) were recorded.     

Molecular phylogeny of Nothoholosticha (Protozoa,Ciliophora, Urostylida) and systematic relationships of the Holosticha-complex

The marine ciliate Nothoholosticha is characterized by having a Holosticha-like ciliature pattern but without migratory frontoterminal cirri. However, its systematic position and its relationship to other members of the Holosticha-complex have not yet been resolved. In order to gain deeper insights into these relationships, the small subunit rRNA (SSrRNA) gene and the rRNA internal transcribed spacer and 5.8SrRNA (ITS1–5.8S-ITS2) region of two marine Anteholosticha species, as well as the ITS1–5.8S-ITS2 region of N. fasciola, were sequenced, and molecular trees (BI, ML, NJ and MP trees) were constructed. Although our analyses failed to conclusively resolve the phylogeny of this assemblage, certain conclusions could be drawn. Firstly, Nothoholosticha is a valid genus that is more closely related to Pseudokeronopsis than to other Holosticha-complex genera. Secondly, sequence analyses and phylogenetic trees of several Anteholosticha species revealed a high molecular diversity, which does not support the monophyly of this genus. Thirdly, the current assignment of certain well-known genera, e.g. Holosticha, Anteholosticha, Apokeronopsis, Parabirojimia, Psammomitra, Diaxonella, Metaurostylopsis and Thigmokeronopsis, to the families Bakuellidae (sensu Berger 2006), Urostylidae (sensu Berger 2006) or Holostichidae (sensu Berger 2006) is challenged by the molecular data presented here. And fourthly, the families Holostichidae and Pseudokeronopsidae (sensu Lynn 2008) are probably paraphyletic, and their systematic assignments await further evaluation.     

pH-Dependent Effects of 2,4-Dinitrophenol (DNP) on Proliferation, Endocytosis, Fine Structure and DNP Resistance in Tetrahymena

ABSTRACT. In view of the importance of external pH on cytotoxic effects of ionizable agents, the pH-dependent effects of 2,4-dinitrophenol (DNP) were investigated. As uncoupler of oxidative phosphorylation. DNP interferes with the proton gradient across the inner mitochondrial membrane. DNP was added to proliferating Tetrahymena pyriformis in media of different initial pH. Effects studied were rates of cell proliferation and endocytosis, and fine structure. Findings correlated with the calculated concentration of undissociated DNP, taking into account that pH changes with time and cell density in Tetrahymena cultures. A linear relationship thus emerged between initial concentrations of undissociated DNP and lengths of the lag preceding cell proliferation. Once resumed, the rate of proliferation corresponded to that of control cells, even in different concentrations of undissociated DNP, presumably indicating an adaptation mechanism. Endocytosis was elevated throughout a wide range of undissociated DNP concentrations with a sharp transition towards inhibition at high DNP concentrations causing lethality with time. Changes in fine structure of DNP-treated cells (mitochondria, peroxisomes, nucleoli) also depended on the concentration of undissociated DNP.     

Effect of quinacrine treatment on the activity of acid hydrolases (phosphatase, N-acetyl-beta-glucosaminidase, glucosidase, galactosidase and esterase) in Tetrahymena.

The effect of phospholipase A2 (PLA2) inhibitor, quinacrine, on the activity of hydrolytic enzymes in Tetrahymena pyriformis homogenate, was investigated. The activity of all of the enzymes studied (acid phosphatase, N-acetyl-beta-glusosaminidase, glucosidase, galactosidase and esterase) was significantly reduced in the presence of quinacrine. Since there are no data on the inhibitory effect of PLA2 and PLA2 influenced metabolic pathways to the hydrolytic enzymes, the direct effect of quinacrine on the hydrolytic enzymes (of Tetrahymena) can be supposed. This is supported by the fact that the other PLA2 inhibitor, 4-bromophenacyl bromide, did not influence phosphatase activity.     

The importance of type species and their correct identification: A key example from tintinnid ciliates (Alveolata,Ciliophora, Spirotricha)

Types and the corresponding rules in the International Code of Zoological Nomenclature are crucial for taxonomy and are meant to provide nomenclatural stability. In the case of neotypification, especially diligent taxonomic work is required to retain continuity. In the present communication, we first outline the main principles of typification and neotypification. We then discuss a critical case, using a current example from the marine planktonic tintinnid genus Tintinnopsis Stein, 1867 (Alveolata, Ciliophora). This diverse and ubiquitous genus is nonmonophyletic, but its revision and the erection of new related genera is currently prevented by the uncertain affiliation of its type species.     

土壤腹毛类纤毛虫四新纪录种

利用非淹没培养皿法、活体观察和蛋白银染色法,研究了青岛的土壤纤毛虫,发现2属4种我国新纪录种:澳洲管膜虫(Cyrtohymena australis Foissner,1995)、四核管膜虫[C.quadrinucleata(DragescoNjine,1971)]、犬牙管膜虫[C.candens(Kahl,1932)]及迟钝澳纤虫(Australocirrus oscitans BlattererFoissner,1988)。文中对该4种纤毛虫的活体形态和纤毛图式进行了观察记述,补充了前人描述的不足,并依据统计学特征与相近种群进行了比较。     

关于《中国植物志》中鹿药(百合科)花特征的纠正

根据野外调查,同时结合文献分析,指出《中国植物志》和Flora of China中关于百合科(Liliaceae)鹿药花部特征描述存在的问题,并提出修改建议。此外,对其学名进行了简要讨论。     

A huge,undescribed soil ciliate (Protozoa: Ciliophora) diversity in natural forest stands of Central Europe

We investigated 12 natural forest stands in eastern Austria for soil ciliate diversity, viz., eight beech forests and two lowland and Pinus nigra forests each. The stands span a wide range of climatic (e.g., 543–1759 mm precipitation, 160–1035 m above sea-level) and abiotic (e.g., pH 4–7.4) factors. Samples were taken twice in autumn and late spring and analysed with the non-flooded Petri dish method. Species were identified in vivo, in silver preparations, and in the scanning electron microscope. A total of 233 species were found, of which 30 were undescribed, a surprising number showing our ignorance of soil ciliate diversity, even in Central Europe. Species number varied highly from 45 (acidic beech on silicate) to 120 (floodplain forest) and was strongly correlated with pH and overall habitat quality, as measured by climate, the C/P quotient (ratio of r-selected colpodean and k-selected polyhymenophorean ciliates), and the proportion of mycophagous ciliate species; multivariate analysis showed further important variables, viz., the general nutrient status (glucose, nitrogen, C/ N ratio) and microbial (urease) activity. The highest species number occurred in one of the two floodplain soils, supporting the intermediate disturbance hypothesis. The three main forest types could be clearly distinguished by their ciliate communities, using similarity indices and multidimensional scaling. Individual numbers varied highly from 135^–1 (lowland forest) to 10,925 ml^–1 (beech on silicate) soil percolate and showed, interestingly, a weak correlation with soil protozoan phospholipid fatty acids. Eight of the 30 new species found and a forgotten species, Arcuospathidium coemeterii (Kahl 1943) nov. comb., are described in detail, as examples of how species were recognized and soil protozoan diversity should be analyzed: Latispathidium truncatum bimicronucleatum, Protospathidium fusioplites, Erimophrya sylvatica, E. quadrinucleata, Paragonostomum simplex, Periholosticha paucicirrata, P. sylvatica, and Australocirrus zechmeisterae.     

日本血吸虫新抗原基因的筛选、克隆、 表达和免疫效果研究

为了寻找日本血吸虫 (Schistosoma japonicum, Sj) 新的疫苗候选基因并进行免疫效果研究,用 Sj 雌虫抗原免疫家兔制备血清,对Sj成虫 cDNA 文库进行免疫筛选,将获得的新基因 ( 命名为Sj-F1, GenBank 登录号为 AY261995) 克隆入原核表达载体 pTWIN1 和真核表达载体 pcDNA3 ,经 PCR 、限制性酶切筛选和鉴定阳性重组子. 将 pTWIN1/Sj-F1 质粒转化大肠杆菌 ER2566,在低温和低 IPTG 浓度下诱导表达可溶性重组融合蛋白 (rSj-F1/intein2),并经 SDS- 聚丙烯酰胺凝胶电泳 (SDS-PAGE) 和蛋白质印迹 (Western blot) 分析鉴定. 将 pcDNA3/Sj-F1 质粒转化大肠杆菌 ER2502 ,大量制备 DNA 疫苗. 用重组融合蛋白和 DNA 疫苗免疫小鼠,末次免疫后 2 周用Sj尾蚴进行攻击感染. 感染后 42 天剖杀冲虫,计算减虫率和减卵率. 感染前采血用 ELISA 法检测抗体. 免疫保护效果测定显示：重组蛋白疫苗以 FCA 作佐剂经皮下免疫和以壳聚糖作佐剂经粘膜免疫分别获得了 28.07%、 24.69% 的减虫率和 48.30% 、 46.38% 的减卵率; DNA 疫苗 (pcDNA3/Sj-F1) 单独免疫获得了 18.47% 的减虫率和 35.06% 的减卵率;用 DNA 疫苗启动免疫后用重组蛋白疫苗经皮下加强免疫,减虫率和减卵率分别提高到了 40.42% 和 56.17%;用 DNA 疫苗启动免疫后用重组蛋白疫苗经黏膜加强免疫,减虫率和减卵率增高更明显,分别提高到了 42.38% 和 62.87%. 结果表明,Sj-F1 重组蛋白疫苗及 DNA 疫苗均可诱导小鼠产生部分抗血吸虫感染的保护力,两者联合免疫保护效果优于单一疫苗.