日本血吸虫子胞蚴超微结构的研究：产孔的形态学证明

本文用透射电镜观察我国大陆品系日本血吸虫子胞蚴前端、收缩区和扩张区的超微结构。发现子胞蚴前端存在体被凹陷区及括约肌的构造,同时可见神经细胞。这个子胞蚴精细分化区可能具有产孔的生理功能,尾蚴从产孔中逸出是非创伤性逸出功能的一种生理适应。并与曼氏血吸虫子胞蚴产孔的超微结构进行了比较。     

日本血吸虫胞蚴期超微结构的初步观察

本文用扫描与透射电镜观察了我国大陆品系37日和45日龄日本血吸虫母胞蚴及其体内未成熟子胞蚴体被的结构。同时观察了取自螺肝组织的62日龄成熟子胞蚴。初次揭示日本血吸虫胞蚴期体被的超微结构,基本上与曼氏血吸虫胞蚴期相似。日本血吸虫母胞蚴和成熟子胞蚴除体被无体棘外,其他很相似。比较未成熟的子胞蚴与未成熟的尾蚴,揭示外质膜由两层结构构成;随后,外层结构溶化消失,而同时出现微绒毛。构成这样母子二代胞蚴及其体内胚胎既相同又有差别,认为与幼虫寄生部位及生殖生理状态有关。     

Ultrastructure of the daughter sporocyst and developing cercaria of Schistosoma japonicum in experimentally infected snails, Oncomelania hupensis hupensis

Ultrastructure of daughter sporocysts and cercariae of Schistosoma japonicum were studied 2 and 4 months after infection of Oncomelania hupensis hupensis. The body walls of daughter sporocysts are similar at all infectious stages. They consist of an external syncytial tegument on a basement membrane, and an internal cellular subtegument surrounding a body cavity containing developing cercariae. The cercariae embryos develop 2 months after infection from germinal balls in the brood chamber of the daughter sporocyst. They are at first enveloped by a primitive epithelium rising from the daughter sporocyst. Four months after infection, the cercariae were almost fully developed and the primitive epithelium had degenerated. The body wall of the cercaria consists of a thin tegument covered by a surface coat of fibrous material and connected to the subtegumental cells by cytoplasmic processes. The matrix of the tegument contains numerous dense bodies, vacuoles, and spines. Two types of sensory structures - uniciliated and multiciliated - are found at the anterior tip of the cercaria. There are five pairs of penetration gland cells of two distinct types differentiated by the morphology of secretory granules. Flame cells are found in both daughter sporocysts and in cercariae. The cilia of the flame cells are characterized by the typical 9 and 2 cilium pattern.     

The development of the primitive epithelium and true tegument in the cercaria of Schistosoma mansoni.

The formation of the final cercarial tegument of Schistosoma mansoni is preceded by that of a so-called primitive epithelium. The primitive epithelium is derived from the tegument of the daughter sporocyst. The final cercarial tegument is formed from peripherally located somatic cells of the cercarial embryo, which expand and coalesce beneath the primitive epithelium. The primitive epithelium degenerates and disappears. The ultrastructure of both epithelia in the course of the development of the cercaria is described in detail. Possible functions are discussed.     

Changes in the monocercus cercomer in the cysticercoid cavity and the hemocoel of the nonspecific host

In order to elucidate the functional role of cercomer in larvae of the monocercus type their transplantation from the specific host Chironomus obtusidens to Gammarus lacustris was conducted. At early stages after the transplantation proceeds an increase in the functional activity of the tegument of follicles of the cercomer followed by their complete destruction in 3 or 4 days. On the surface of the exocyst membrane an adhesion of the host's haemocytes occurs, which becomes more distinct in 3-4 days when the process acquires a character of local encapsulation. Within the same period, in the places of haemocytes aggregation, a local resorption of the exocyst external membrane takes place. Later intensification of the host response to transplant is associated with the destruction of follicles of the cercomer. In one case the occurrence of follicles of the cercomer in the cavity of cysticercoid was observed that is caused by the microbe affection of the latter. In the zone of contact of the tegument of scolex and neck with follicles of the cercomer an increased secretion (the microapocrine type) of the tegument, disturbance of the microvillous tegument of the cercomer's follicles and their destruction are observed. Incompatibility of the tegument of definitive departments and cercomer, which arises during differentiation of larvae, is supposed to affect the formation of scolex invagination in the evolution of larvae of Hymenolepis.     

Cytoskeletal features of the syncytial epidermis of the tapeworm Hymenolepis diminuta

A hallmark feature of parasitic platyhelminths is a cytoarchitecturally unusual syncytial epidermis composed of a peripheral layer of continuous cytoplasm (the ectocytoplasm) connected to underlying nucleated cell bodies by small cytoplasmic bridges. The helminth epidermis, or tegument, plays important roles in protection and nutrient acquisition; cestodes, in fact, completely lack a gastrointestinal tract and absorb all nutritive material through the tegument. Perhaps not surprisingly, the cestode tegument bears certain resemblances to the mucosal epithelium of the vertebrate small intestine, including the possession of a microvillous brush border upon the surface of the ectocytoplasm. In contrast to the intestinal epithelial cell, however, very little is known concerning the nature and organization of the cytoskeleton within the helminth epidermis. Therefore, a number of different microscopical preparative techniques were used to examine the tegument of the tapeworm Hymenolepis diminuta for the presence and distribution of microfilaments, intermediate filaments, and microtubules. It was found that both actin-containing microfilaments and intermediate-sized filaments are present but are restricted to specific locations along the plasmalemmae of the ectocytoplasm. In contrast, microtubules are found throughout the tegument, and are concentrated in the supranuclear regions of the perikarya and in the cytoplasmic bridges interconnecting the perikarya and ectocytoplasm. Unlike brush borders of most other epithelia, the cestode epidermal brush border lacks a filamentous terminal web and is instead associated with microtubules. A network of fine filaments, 5-8 nm in diameter but distinct from actin-containing microfilaments, runs throughout the ectocytoplasm and appears to interlink tegumental vesicles. These fine filaments may represent the primary "skeletal" system responsible for maintaining the structure of the tegumental cytoplasm.     

Regional specializations of the tegument of the metacercaria of Timoniella imbutiforme

The ultrastructure of the spinous body tegument of the metacercaria of Timoniella imbutiforme (Molin, 1859) has recently been described. Other regions of the metacercarial tegument, including those of the oral sucker, pharynx, and nephridiopore, demonstrate considerable specializations. The oral sucker tegument had an aspinous outer syncytial layer that possessed a pimpled apical surface as well as enclosing two types of secretory bodies. The pharyngeal tegument likewise lacked spines, but possessed only one type of secretory body, and a smooth but folded outer surface. The nephridiopore tegument, however, showed the greatest degree of specialization possessing a single type of secretory body specific only to this region of the tegument. Also associated with the syncytium here was a prominent long filamentous glycocalyx, and microtubules which were observed for the first time in this region of the tegument.     

Scanning electron microscopy of the vitelline membrane of the hen ovum

Manual removal of the perivitelline layer overlying the animal pole (AP) reveals three morphologically distinct regions of the vitelline membrane (VM). (1) The central germinal region is 600-800 micron in diameter and is densely populated with pleomorphic microvillous projections. (2) The periblastic region, which also exhibits microvillous projections, is 250-550 micron wide and consists of numerous (80-120) lacunae that are 10-60 micron in diameter and up to 20 micron in depth. (3) At the outer periblastic region, the microvillous projections are less numerous. In the vegetal hemisphere, the VM has few projections and occasionally is discontinuous.     

Morpho-anatomic and functional sectorization of Schistosoma mansoni daughter sporocysts

During the larval development of S. mansoni in the snail host, morpho-anatomic changes occur in the daughter sporocyst by a sectorization of this larval stage. Three sectors can be distinguished: an anterior zone with a well-differentiated birth pore; dilated zones containing the developing cercariae; constricted zones without cercarial embryo. The photonic and electronic microscopical study shows variations in the tegumental structure of these sectors. This evolution of the daughter sporocysts is discussed in relation with the dynamics of larval stages and the replication process of sporocysts.     

Fasciola hepatica: tegumental alterations in adult flukes following in vitro and in vivo administration of artesunate and artemether

The tegumental changes in adult Fasciola hepatica induced by artemether and artesunate were assessed utilizing scanning electron microscopy (SEM). F. hepatica were incubated with artemether and artesunate for 48h at a concentration of 10microg/ml in the absence or presence of haemin. For the latter experiment both, a triclabendazole-resistant and sensitive F. hepatica isolate were used. For the in vivo studies rats were treated with single 200mg/kg oral doses of artemether and artesunate and flukes recovered from the bile ducts after 24-96h. SEM analysis of the flukes incubated in the presence of the drugs without haemin showed only minor and localized damage of the tegument. In the presence of haemin extensive tegumental damage, including sloughing, blebbing and eruptions, particularly in the ventral and dorsal mid-body and tail region, was evident. No difference in the extent of damage could be observed between artemether and artesunate and between the triclabendazole-resistant and non-resistant flukes. After 24h in vivo disruption of the tegument was evident in the artemether-treated flukes, and the damage increased in severity 48-72h post-treatment. Sloughing, swelling and extensive furrowing of the tegument was observed in several flukes, in particular in the tail region and the ventral apical cone region. In the artesunate treatment, tegumental damage was evident after 72h, but seemed slightly less pronounced when compared to the artemether-treated specimens examined at the same time point. Concluding our experiments confirm that artemether and artesunate are potent fasciocidal drugs and the tegument of adult F. hepatica appears to be a target for the action of these drugs.     

Hymenolepis diminuta: ultrastructural observations on complement-mediated tegumental lysis and destrobilation of 4-day-old worms in vitro

Changes in the ultrastructure of the tegument and subtegumental cells of 4-day-old Hymenolepis diminuta were studied in vitro in 50% fresh normal rat serum over a 5-h period and compared with heat-inactivated serum and saline controls. First, membrane-bound vesicles accumulate above the microthrix-border. After 30–40 min large vacuoles, which may contain membranous elements, appear in the tegument at a time when the surface of the young strobila is virtually denuded of the microthrix-border. With prolonged incubations there are subtegumental secretory inclusions with dark, enveloping cytoplasm in the tegument and finally the apical plasma membrane, together with the majority of the matrix, is lost. The disrupted portion of the worm is abruptly demarcated from the comparatively intact scolex/anterior neck region by a constriction. Even after 5 h incubation there is no evidence of loss of tegumental matrix components from regions anterior to the constriction but the neck region shows a significant denudation of the microthrix layer and the tegument contains numerous inclusions. The scolex tegument only showed little evidence of loss of membrane from the surface. Possible mechanisms for the avoidance of complement-mediated lysis in the anterior region are discussed.     

Cytomorphological characteristics of the tegument of daughter larvae in the cestode Taenia crassiceps

At the early stages of asexual breeding, the daughter larvae of T. crassicaps display destructive changes of the tegument due to its hyperfunction. The reparation of tegumental cytons is followed by the appearance of specific "brushes" of fibrillar material on the karyolemma of their nuclei. They are supposed to occur as a result of protein synthesis necessary for the reconstruction of cellular structures, mainly, of the nucleus. The abundant synthesized fibrillar falls into decay being transformed into bodies containing crystals which are transported to the superficial synthytium to be eliminated there.     

Schistocephalus solidus: pinocytosis by the plerocercoid tegument

The possibility of pinocytosis occurring in the tegument of the plerocercoid of Schistocephalus solidus has been investigated by morphological and experimental methods. Electronmicroscopic study showed that the outer syncytial tegument contained numerous electronlucid vesicles. These vesicles had two gradients, the number of vesicles decreasing from the outer canopy region to the inner canopy and from the apical to the basal plasma membrane for any particular region of tegument. A variety of morphological modifications of the apical plasma membrane very similar to those which have been accepted as evidence of pinocytosis in other tissues were present. In vitro studies using horseradish peroxidase, ruthenium red, and lanthanum nitrate showed that all three tracers are taken up by the tegument into membrane-limited vesicles. Vesicles which contained ruthenium red occurred at the base of the tegument within a 5-min incubation period, and their contents appeared to be released into the underlying interstitial material by exocytosis.     

鱊头槽绦虫原尾蚴皮层的超微结构观察

利用透射电镜观察了头槽绦虫 (Bothriocephalusacheilognathi)原尾蚴皮层的超微结构。头槽绦虫原尾蚴的皮层为典型的合胞体结构。皮层表面有浓密的微毛与少量的突起。与成虫相比 ,原尾蚴的微毛长而且粗 ,呈现单态性 ,推测在进入终末宿主的过程中有脱落和再生现象。突起分布在头部与体侧 ,含有较少的电子致密颗粒。在原尾蚴皮层细胞质中观察到三种腺细胞的分泌过程 ,即顶分泌、外分泌和微分泌过程。原尾蚴身体后部观察到尾钩和穿刺腺管道的开口。在核周区和纤维层下可见发育较好的环肌与纵肌。本文还对原尾蚴皮层突起的功能进行了探讨。     

Fine Structure of Cephalic Sense Organs in Heterodera glycines Males

Cephalic sense organs of Heterodera glycines males were examined in detail by electron microscopy. Each amphid basically consists of an amphidial gland, a nerve bundle, and an amphidial duct. The amphidial gland consists of a microvillous region, and laterally is closely associated with a large secretory cell. The nerve bundle penetrates the microvillous region, and further anteriorly individual nerve processes (dendrites) separate from one another, thus forming a sensilla pouch which is enveloped by the microvillous region of the gland. Anterior to the pouch, the cilia-like dendrites converge as they enter and eventually terminate in the amphidial duct. Heterodera glvcines males have the innervation basis for a full complement of sixteen papillae, although surface manifestations are present for only six minute inner labial papillae. In addition, four outer labial and four cephalic receptors terminate beneath the surface, and another two dendrite pairs end further posteriorly beneath the basal plate of the cephalic framework. Papillary receptors which terminate beneath the surface are probably mechanoreceptive, whereas inner labial papillae have pore-like openings to the exterior and may be chemoreceptive. Amphids and papillae of H. glycines are fundamentally similar to those of Meloidogyne incognita, although certain striking differences exist.     

First report of alloparental care in the Key Largo woodrat (Neotoma floridana smalli)

Here we report the first observation of alloparental care in a subadult female Key Largo woodrat. Maternal and other behaviors were scored for the subject dam and her subadult daughter at the birth of the dam’s second litter and the following 30 days. After assisting with birth, the subadult exhibited maternal behaviors and remained in close proximity to the pups. The rate of maternal care behaviors observed in the subject dam was lower than in solitary dams without an alloparent present. However, the total maternal care score for the dam and subadult combined approached the average score for single dams.     

Volume growth of daughter and parent cells during the cell cycle of Saccharomyces cerevisiae a/alpha as determined by image cytometry.

The pattern of volume growth of Saccharomyces cerevisiae a/alpha was determined by image cytometry for daughter cells and consecutive cycles of parent cells. An image analysis program was specially developed to measure separately the volume of bud and mother cell parts and to quantify the number of bud scars on each parent cell. All volumetric data and cell attributes (budding state, number of scars) were stored in such a way that separate volume distributions of cells or cell parts with any combination of properties--for instance, buds present on mothers with two scars or cells without scars (i.e., daughter cells) and without buds--could be obtained. By a new method called intersection analysis, the average volumes of daughter and parent cells at birth and at division could be determined for a steady-state population. These volumes compared well with those directly measured from cells synchronized by centrifugal elutriation. During synchronous growth of daughter cells, the pattern of volume increase appeared to be largely exponential. However, after bud emergence, larger volumes than those predicted by a continuous exponential increase were obtained, which confirms the reported decrease in buoyant density. The cycle times calculated from the steady-state population by applying the age distribution equation deviated from those directly obtained from the synchronized culture, probably because of inadequate scoring of bud scars. Therefore, for the construction of a volume-time diagram, we used volume measurements obtained from the steady-state population and cycle times obtained from the synchronized population. The diagram shows that after bud emergence, mother cell parts continue to grow at a smaller rate, increasing about 10% in volume during the budding period. Second-generation daughter cells, ie., cells born from parents left with two scars, were significantly smaller than first-generation daughter cells. Second- and third-generation parent cells showed a decreased volume growth rate and a shorter budding period than that of daughter cells.     

Schistosoma mansoni: development of the cercarial glycocalyx

The development of the cercarial glycocalyx of Schistosoma mansoni was studied by transmission electron microscopy and immunofluorescence light microscopy employing antibodies raised against extracted and chromatographed glycocalyx. By electron microscopy, cercariae present in the brood chamber of daughter sporocysts were surrounded by an electron-dense granular and fibrillar matrix. This material appeared structurally distinct from the glycocalyx which was coarsely fibrillar and located only on the surface of organisms that had developed a final tegument. The thickness of the glycocalyx apparently increased with the maturation of the tegument, since teguments that had many spines also had the thickest glycocalyx. Immunofluorescent staining of frozen sections of infected snail hepatopancreas showed that glycocalyx antigens were present on the surface of the cercariae and not in the matrix within the brood chamber or in snail tissues. Immunofluorescent staining of isolated larval cercariae showed staining of some but not all parasites with partially elongated tails. These studies suggest that the glycocalyx develops late in cercarial development (late in Stage 6 or in Stage 7 of Cheng and Bier), is made by the cercariae themselves, and is not a product of either the sporocyst wall cells or snail hepatopancreas.     

长尾蚴吸虫幼虫在菲律宾蛤仔的寄生部位及其组织化学的研究

本文报道长尾蚴吸虫幼虫在菲律宾蛤仔的寄生部位及其组织化学的研究,长尾蚴吸虫幼虫主要寄生于哈仔的生殖腺组织中,仅少部分进入生殖腺附近的消化腺,鳃等。受感染严重的蛤仔,其生殖腺的滤泡等组织被虫体占满并被耗尽,长尾蚴吸虫子胞蚴的胞壁及尾蚴的皮导主要含碱性蛋白质和粘蛋白,子胞蚴的胚球,尾蚴的吸盘,生殖原基,单细胞腺体主要ＤＮＡ阳性物质,排泄囊含丰富的碱性蛋白质;尾蚴体的中部含有呈颗粒团的粘蛋白和抗淀粉酶的     

Carbohydrate Mimicry of the Parasite in the <Emphasis Type="Italic">Himasthla elongata</Emphasis> (Trematoda: Echinostomatidae)—<Emphasis Type="Italic">Littorina littorea</Emphasis> (Mollusca: Prosobranchia) System

Using labeled lectins, a comparative study of the surface of tegument of Himasthla elongata at different stages of development cycle (daughter rediae, cercariae, and metacercariae) and glycocalyx of plasma membranes of hemocytes of molluscs Littorina littorea and Mytilus edulis that are the first and second intermediate hosts of this trematode species, respectively, has been carried out. It is found that in the course of the development cycle of the parasite there occurs a change of the set of terminal sacharides of the glycocalyx of the H. elongata tegument surface as well as differences are revealed in the pattern of binding of three out of five tested lectins with hemocytes of blue mussel and periwinkle. At the same time, the presence of similar carbohydrate determinants on the surface of hemocytes of L. littorea and daughter rediae of H. elongata is shown. The established similarity in composition of glycocalyx of hemocytes of the mollusc and the trematode parthenitae is, most likely, a result of coevolution of the parasite and the host and is of the adaptive nature. Use of the mechanism of the carbohydrate mimicry by parthenitae of this species allows them to avoid attacking by effector cells of the internal defense system of the mollusc-host.__________Translated from Zhurnal Evolyutsionnoi Biokhimii i Fiziologii, Vol. 41, No. 2, 2005, pp. 114–118.Original Russian Text Copyright © 2005 by Iakovleva, Gorbushin.