三种纤毛虫细胞内酸性磷酸酶的定位观察

利用电镜酶细胞化学方法,对大尾柱虫(Urostylagrandis)、冠突伪尾柱虫(Pseudourostylacristata)、魏氏拟尾柱虫(Paraurostylaweissei)三种纤毛虫细胞内的酸性磷酸酶(ACPase)进行了定位观察。发现在它们体内ACPase的阳性反应颗粒除分布在消化腔隙内独立的泡状结构上和被消化的食物组织中外,在细胞质中的小圆泡内也有分布。所得结果证实了消化腔隙和小圆泡都具有消化功能。作者推测消化腔隙主要消化食物,小圆泡则可能与自身物质(如衰老胞器等)的消化有关;并推测这种消化腔隙与小圆泡共存是纤毛虫消化胞器由食物泡演化为消化腔隙系统的中间形式。此外,在大尾柱虫纤毛内部观察到非溶酶体性的ACPase。     

食物在贻贝棘尾虫(原生动物,纤毛虫)细胞内的消化过程

利用免疫荧光显微术、透射电镜及电镜酶细胞化学技术对食物在腹毛类纤毛虫-- 贻贝棘尾虫细胞内的消化过程进行了追踪观察.食物在进入消化腔隙的初期,外面包被着一层膜结构,但此外层膜很快被消化而消失;尔后食物在贻贝棘尾虫体内的消化过程可分为两种方式:一种方式为直接在消化腔隙内完成,此过程同1982年Kaul et al.和Das s et al.的报道;而另一部分食物的消化过程表现为食物逐步向细胞质内突入,以致最终完全被细胞质包围形成一被膜包裹的圆泡状结构;在此过程中,细胞质边缘突出众多含有酸性磷酸酶等物质的小囊泡结构,小囊泡与食物融合,其内的酸性磷酸酶等水解酶释放出来, 食物的表膜逐渐瓦解,个体逐渐变小,最终消化后的营养物质被释放到细胞的消化腔隙内; 这种食物通过胞口与胞咽达消化腔隙后,再由细胞质将其包裹形成的食物泡,我们称之为" 后成食物泡".这两种消化方式与贻贝棘尾虫的消化胞器为一腔隙结构相适应[动物学报 5 4(3):510-516,2008].     

纤毛虫念珠异列虫射出胞器的超微结构观察

应用扫描电镜术和透射电镜术显示,纤毛虫念珠异列虫(Anteholosticha monilata)的射出胞器早期发生在细胞质深处,附近有不同类型的囊泡结构。成熟后射出胞器向表膜迁移,结构由不同电子密度片层的体部、结晶状的中心轴杆部和多层膜的帽部组成。受外界刺激时胞器冲破皮层射出,形态呈"蘑菇"状。据上述观察结果推测:该射出胞器具有防御作用,它可能起源于高尔基体活动产生的小泡;在亲缘关系较近的纤毛虫中,其射出胞器可能具有相似的分化特征。     

魏氏拟尾柱虫休眠包囊及其细胞器超微结构的观察

为研究纤毛虫休眠状态下细胞的分化及其胞器的特征,本文以透射电镜术显示,魏氏拟尾柱虫（Ｐａｒａｕｒｏｓｔｙｌａｗｅｉｓｓｅｉ）休眠包囊中,颗粒层壁内有小泡,表膜位置偶见小泡样结构,大部分线粒体以多个相互聚集在一起,自噬泡将线粒体等胞器包裹在内经历消化过程,细胞内膜系统十分发达。作者推测,颗粒层及表厝小泡可能是休眠细胞经由表膜进行物质交换的结构,自噬泡消化现象可能是细胞中物质和能量来源的主要途径。     

松花江四种纤毛虫的形态学研究

利用活体观察, 氨银染色和蛋白银染色的方法, 对采自松花江的4种广布种纤毛虫: 贻贝棘尾虫Stylonychia mytilus (Müller, 1773) Ehrenberg, 1830、鬃异源棘尾虫Tetmemena pustulata (Müller, 1786) Eigner, 1999、尾草履虫Paramecium caudatum (Ehret, 1963)和刚毛榴弹虫Coleps hirtus的形态学进行研究。依据新采集种群, 对4个种活体形态和纤毛图式特征, 以及鬃异源棘尾虫的发生学特征进行补足性描述。研究对贻贝棘尾虫做了重要重描述, 可清晰观察到该种蛋白银染后6列背触毛的排列特征、左右缘棘毛数的具体统计, 结果发现, 贻贝棘尾虫哈尔滨种群与前人报道的种群相比, 形态特征基本吻合, 区别在于前者小膜数目多于后者。鬃异源棘尾虫哈尔滨种群与漠河种群相比, 形态和纤毛图式方面十分相似, 发生模式基本一致, 细微区别在于前者左右缘棘毛数目普遍多于后者, 前者虫体大小波动范围较小, 小核数目多于后者。尾草履虫哈尔滨种群与其青岛种群相比, 二者显著差异在于收集管数目前者比后者少。本文榴弹虫与前人报道的刚毛榴弹虫种群相比, 盔板带的数目、排列方式、窗口类型等形态学基本一致, 细微区别在于本文报道种群体型普遍较前人报道种群略大。     

浮萍棘尾虫Adss基因克隆及表达载体构建

为研究腺苷酸基琥珀酸合成酶(Adenylosuccinate synthase, 简称Adss)在原生动物中的作用, 对浮萍棘尾虫(Stylonychia lemnae)腺苷酸基琥珀酸合成酶Adss基因进行PCR克隆, 获得全长为1396 bp左右的序列, 其编码458个氨基酸, 保守结构域含有一个P-loop NTPase结构域, 蛋白结构预测Adss蛋白是由α-螺旋、β-折叠和无规卷曲组成的复杂结构。多重序列比对和系统进化分析显示浮萍棘尾虫Adss蛋白与第四双小核草履虫、多子小瓜虫和嗜热四膜虫等纤毛虫同源性较高。通过克隆浮萍棘尾虫Adss基因的启动子序列, 并采用增强型绿色荧光蛋白EGFP作为报告基因, 构建了启动子序列调控的表达载体pEGFP-N1-Adss, 通过转染细胞确定浮萍棘尾虫Adss蛋白定位于细胞质中。文章报道了原生动物纤毛虫Adss基因, 证实浮萍棘尾虫Adss蛋白属于典型的真核生物Adss, 为进一步揭示Adss在单细胞真核生物中的作用机制提供了新的参考。     

沼泽瘦尾虫皮层纤毛器及其附属微管的观察

应用扫描电镜技术、荧光紫杉醇直接荧光标记显示了腹毛目纤毛虫沼泽瘦尾虫(Uroleptus limnetis)的细胞形态和皮层纤毛器的组成模式,以及皮层口围带、额腹横棘毛、左右缘棘毛等纤毛器微管和纤毛器附属微管的建构特征,可为进一步阐明瘦尾虫类纤毛虫的形态学及其系统发育研究提供基础资料。     

多节核棘尾虫无小核系虫体的初步研究

纤毛虫细胞具有大核和小核之分。多年来认为大核控制当代无性系细胞的全部代谢活动,小核只在有性生殖时起产生两性原核和受精的作用,从而更新无性系。近年来的研究证明,小核对当代无性系细胞的皮层形态及形态发生也起相当重要的作用。这此工作是在草履虫上(Ng等,1981,1984,1987)和贻贝棘尾虫上(史新柏等,1983,1990;Lu等,1989)完成的。但小核对大核的影响在任何纤毛虫上尚未见报道。笔者用所获得的新种多节核棘尾虫     

魏氏拟尾柱虫腹皮层纤毛器微管胞器的形态及形态发生

应用荧光紫杉醇直接荧光标记和抗α-微管蛋白抗体免疫荧光标记显示,魏氏拟尾柱虫(Paraurostyla weissei)腹面皮层纤毛器微管胞器由口围带、波动膜、额腹横棘毛和左右缘棘毛等纤毛器微管、纤毛器基部附属微管等组成。其中口围带基部微管包括小膜托架、小膜附属微管;额腹横棘毛和左右缘棘毛基部附属微管包括前纵微管束、后纵微管束和横微管束,它们由各自的纤毛器基部向皮层细胞质不同方向发射,形成腹皮层表面下微管网。结果表明,魏氏拟尾柱虫的纤毛器骨架、纤毛器附属结构也是一类以微管蛋白为基本成分的微管胞器,其中缘棘毛基部附属微管具有不同于其他纤毛虫(例如棘尾虫)中所观察到的同种微管胞器的建构特征。形态发生中,前仔虫口围带在老结构位置形成,其结构建成与部分老口围带的更新有关;老缘棘毛的结构物质对新的左、右缘棘毛的发生可能具有定位作用及物质贡献,但此后新的左、右缘棘毛列分别在老缘棘毛的右侧形成,而并非是在老缘棘毛位置分化的。在有些细胞中,新的左缘棘毛左侧另有一列棘毛,这可能是形态发生中老的左缘棘毛退化不完全产生的。     

兰州八里镇蔬菜基地春季土壤纤毛虫群落特征

2005年4月至2007年6月,用活体观察和固定染色方法对兰州八里镇蔬菜基地春季土壤纤毛虫的群落特征进行了研究.共鉴定出土壤纤毛虫67种,其中包括2个未定名种和14个中国土壤纤毛虫新纪录种.在八里镇蔬菜基地的土壤中鉴定到纤毛虫34种,隶属于3纲、7目、17科、18属;下毛目、肾形目、前口目为其优势类群;吻吴氏虫(Woodruffia rostrata)、近亲游仆虫(Euplotes afflnls)、苔藓圆纤虫(Strongylidium muscorum)、鱼形瘦尾虫(Uroleptus piscis)、似膜袋虫(Cyclidium simulans)等为其优势种.在对照样点(兰山)中鉴定到纤毛虫49种,隶属于3纲、9目、23科、30属;下毛目、盾纤目、肾形目和篮口目为其优势类群;苔藓游仆虫(E.muscicola)、近缘殖口虫(Gonostomum affine)、皮速体虫(Tachysoma pellionella)、食藻斜管虫(Cyclidium simulans)等为其优势种.八里镇蔬菜基地和对照样点的Gleason-Margalef物种多样性指数分别为3.672、5.393;与对照样点土壤纤毛虫的群落Jaccard相似系数为0.239.结果表明,八里镇蔬荣基地春季土壤纤毛虫的物种多样性与对照样点相比有所降低,其群落结构简单化.     

伊曲康唑对皮肤癣菌形态学影响研究

目的观察皮肤癣菌在伊曲康唑作用下的形态学变化。方法应用美国CLSI制订的标准M38-A方案进行伊曲康唑对皮肤癣菌的体外药敏试验,测定最小抑菌浓度（MIC）,将伊曲康唑作用前后的皮肤癣菌分别制成标本,在光学显微镜、扫描电子显微镜和透射电子显微镜下观察形态学变化。结果伊曲康唑作用于皮肤癣菌后,在光学显微镜下菌丝变得弯曲、短粗,顶端和局部出现膨大;扫描电子显微镜下菌丝变得弯曲、短粗、干瘪,顶端和局部出现膨大,有不规则分支,表面粗糙,有大小不等的凹陷;透射电子显微镜下菌丝变得皱缩,有凹陷,双层细胞壁结构消失或不完整,细胞膜不连续,皱缩细胞膜和细胞壁之间及胞浆内出现许多小的高电子密度颗粒,细胞器也变得不清晰。结论伊曲康唑使皮肤癣菌的形态发生明显变化。     

Electron microscopic observation of the invasion process of Cryptosporidium parvum in severe combined immunodeficiency mice

Cryptosporidium parvum mainly invades the intestinal epithelium and causes watery diarrhea in humans and calves. However, the invasion process has not yet been clarified. In the present study, the invasion process of C. parvum in severe combined immunodeficiency (SCID) mice was examined. Infected mice were necropsied; the ilea were double-fixed routinely and observed by scanning and transmission electron microscopy. In addition, the microvillus membrane was observed by ruthenium red staining. Scanning electron micrographs showed elongation of the microvilli at the periphery of the parasite. The microvilli were shown to be along the surface of the parasite in higher magnification. Transmission electron microscopy confirmed that the invading parasites were located among microvilli. Parasites existed in the parasitophorous vacuole formed by the microvillus membrane. The parasite pellicle attached to the host cell membrane at the bottom of the parasite, and then the pellicle and host cell membrane became unclear. Subsequently, the pellicle became complicated and formed a feeder organelle. In addition, invasion of the parasite was not observed in either a microvillus or the cytoplasm of the host cell. Therefore, C. parvum invades among microvilli, is covered with membranes derived from numerous microvilli, and develops within the host cell.     

Paraneuronal pseudobranchial neurosecretory cells in scorpion catfish Heteropneustes fossilis: an environment scanning electron microscope and transmission electron microscope study

Yadav, L., Sengar, M., Zaccone, D. and Gopesh, A. 2011. Paraneuronal pseudobranchial neurosecretory cells in scorpion catfish Heteropneustes fossilis: an environment scanning electron microscope and transmission electron microscope study. —Acta Zoologica (Stockholm) 00 : 1–8. Pseudobranchial neurosecretory system (PNS), found in the gill region of certain groups of teleosts, falls under the category of the ‘diffuse neuroendocrine system’ (DNES). The cells belonging to the system share morpho‐functional features with the paraneuronal cells observed in respiratory tract and airway surfaces of higher vertebrates. On the basis of the experimental observations, a role in condition of hypoxia has been recorded for this system. In an attempt to elucidate the ultrastructure of pseudobranchial neurosecretory cells, present investigation was undertaken using environment scanning electron microscope (ESEM) and TEM in an air‐breathing catfish, Heteropneustes fossilis. The external morphology of PNS under ESEM appeared as a mass of cells supplied with nerves and blood capillaries. Each cell mass is made up of numerous pear‐shaped neurosecretory cells, confirmed by neurosecretion‐specific acid violet stain. The TEM investigation of the cells revealed the presence of different sizes of dense‐cored vesicles in the cytoplasm, which was observed as granular cytoplasm under light microscope. Presence of large number of mitochondria in the cytoplasm confirmed active involvement of these cells in the physiology of fishes. Although lacuna prevails regarding the exact function of this system of fish, its probable role in hypoxic condition and surfacing behavior are speculated.     

Pseudomonas pellicle in disinfectant testing: electron microscopy, pellicle removal, and effect on test results.

Pseudomonas aeruginosa ATCC 15442 is a required organism in the Association of Official Analytical Chemists use-dilution method for disinfectant efficacy testing. When grown in a liquid medium, P. aeruginosa produces a dense mat or pellicle at the broth/air interface. The purpose of this investigation was to examine the pellicle by scanning electron microscopy, to evaluate three pellicle removal methods, and to determine the effect of pellicle fragments on disinfectant efficacy test results. The efficacies of three methods of pellicle removal (decanting, vacuum suction, and filtration) were assessed by quantifying cell numbers on penicylinders. The Association of Official Analytical Chemists use-dilution method was used to determine whether pellicle fragments in the tubes used to inoculate penicylinders affected test results. Scanning electron micrographs showed the pellicle to be a dense mass of intact, interlacing cells at least 10 microns thick. No significant differences in pellicle removal methods were observed, and the presence of pellicle fragments usually increased the number of positive tubes in the use-dilution method significantly.     

Transmission and Scanning Electron Microscopy of the Evaginative Budding Process in Heliophrya sp. (Ciliata,Suctoria)1

ABSTRACT. A sessile, tentacle-bearing protozoon, Heliophrya sp. (Suctoria, Ciliata), reproduces asexually by evaginative budding to form a ciliated swarmer, which begins metamorphosis to the adult form within 30 min of its release from the parent cell. Morphological features of embryogenesis were investigated using transmission and scanning electron microscopy and found to correspond, with certain exceptions, to the few previous reports concerning evaginative budding in suctorians. Following invagination of a portion of the pellicle to form an embryonic cavity within the parent cell, numerous kinetosomes, apparently formed de novo, organize into rows which surround the embryonic cavity and eventually develop cilia that project into the cavity. When the cavity is complete, its walls are extruded through an opening in the parent cell surface. Parent cell cytoplasm streams into the incipient swarmer, thus supplying it with at least the minimum requirement of all cytoplasmic organelles. The ciliated swarmer remains attached to its parent cell for several minutes before it detaches. A complete pellicle is formed in both parent and swarmer prior to detachment. The numerous mitochondria underlying the parent cell pellicle in the vicinity of the attachment area suggest that cross wall formation is an energy-dependent process.     

Pseudomonas pellicle in disinfectant testing: electron microscopy, pellicle removal, and effect on test results

Pseudomonas aeruginosa ATCC 15442 is a required organism in the Association of Official Analytical Chemists use-dilution method for disinfectant efficacy testing. When grown in a liquid medium, P. aeruginosa produces a dense mat or pellicle at the broth/air interface. The purpose of this investigation was to examine the pellicle by scanning electron microscopy, to evaluate three pellicle removal methods, and to determine the effect of pellicle fragments on disinfectant efficacy test results. The efficacies of three methods of pellicle removal (decanting, vacuum suction, and filtration) were assessed by quantifying cell numbers on penicylinders. The Association of Official Analytical Chemists use-dilution method was used to determine whether pellicle fragments in the tubes used to inoculate penicylinders affected test results. Scanning electron micrographs showed the pellicle to be a dense mass of intact, interlacing cells at least 10 microns thick. No significant differences in pellicle removal methods were observed, and the presence of pellicle fragments usually increased the number of positive tubes in the use-dilution method significantly.     

Scanning and transmission electron microscopy of human ejaculate spermatozoa with special reference to their abnormal forms

Summary Spermatozoa from fertile and infertile human ejaculates were observed under the scanning electron microscope. A parallel study of sections was performed by transmission electron microscope.The normal head shows under the scanning electron microscope vesicular elevations in the region of the acrosome and a smooth and rigid appearance corresponding to the postnuclear cap whose occurrence is confirmed under the transmission electron microscope. Immediately anterior to this cap a shallow furrow transverses the head. Duplicated, unusually large or small and deformed heads are found under the scanning electron microscope. Most of these abnormal heads show no surface structure suggesting an acrosome.The neck and middle piece are occasionally, though frequently in abnormal spermatozoa, covered by a cytoplasmic droplet. Otherwise, the mitochondrial sheath is recognized under the scanning electron microscope as a beaded thickening in the middle piece. The lack of mitochondria is manifested by a smooth middle piece thinner than the principal portion. Transmission electron microscopy of sections reveals various types of anomalies in the number of cores, core filaments and mitochondria embedded in the cytoplasmic droplets.Abnormalities in the principal portion of the tail such as duplication, unusual thickness and length are shown under the scanning electron microscope.The investigation indicates that scanning electron microscopy is suited for the clinical as well as cytological examination of human ejaculate spermatozoa.     

Ultrastructure de la Grégarine Callynthrochlamys phronimae Frenzel; Étude Comparée de son Noyau avec Celui de Thalicola salpae (Frenzel) (Eugregarina)

SYNOPSIS. The structure of the gregarine Callynthrochlamys phronimae has been studied with the electron microscope. The cuticular complex is not different from those previously described in other species of gregarines. It is composed of 2 layers of different thickness delimited by 3 unit membranes and constitutes series of oblique folds at the surface of the deutomerite. Longitudinal rods of dense material surrounded by a slight pellicle are seen under the cuticle. Pinocytic vacuoles are present under the surface of the gregarine. Cytoplasmic organelles include mitochondria, Golgi complexes, endoplasmic reticulum, vacuoles and dense bodies from different sizes. There is a connection between the different features of the cytoplasm in the protomerite and deutomerite and the corresponding cuticular organization.
A characteristic feature of the species is the peculiar differentiation of the nuclear membrane. The nucleus is surrounded by a typical double membrane of which the inner one has a dense fibrillar layer apposed to it. In mature trophozoites, tubular expansions without inner layer arise from the double membrane; the same type of nuclear membrane occurs in another species, Thalicola salpae.