Ultrastructural Study of Schizogony in Eimeria callospermophili*

SYNOPSIS The development of 1st generation schizonts of Eimeria callospermophili was studied with cell cultures and with experimentally infected host animals, Spermophilus armatus. Sporozoite-shaped schizonts each had 5-10 nuclei and all of the organelles of the sporozoite; each nucleus had a nucleolus and an associated Golgi apparatus. In stages immediately preceding merozoite formation, an intranuclear spindle apparatus with conical polar areas were observed near the outer margin of each nucleus. Two centrioles, each having 9 single peripheral tubules and one central tubule, were observed near each pole in some specimens. Merozoite formation began internally, with anlagen of 2 merozoites developing near each nucleus. The inner membrane of the merozoites first appeared as 2 dense thickenings adjacent to the polar cones and centrioles; subpellicular microtubules appeared simultaneously. Two anterior annuli and the conoid formed between the 2 thickenings. Vesicles, possibly of Golgi origin, were located next to the forming inner membrane. As the forming merozoites underwent elongation, a rhoptries anlage, a Golgi apparatus, refractile bodies, and mitochondria were incorporated into each. Sporozoite-shaped schizonts with merozoite anlagen transformed into spheroid or ovoid schizonts; at this time the conoid, rhoptries, micronemes, and the inner membrane of the pellicle gradually disappeared; several small refractile bodies were formed from the larger one. When development was about 1/3 complete, the immature merozoites began to grow outward from the surface of the schizont. In this phase of development, the single surface membrane of the schizont became the outer membrane of the merozoite's pellicle, and additional organelles, including the nucleus, were incorporated. Finally, the merozoites became pinched off, leaving a residual body. Development in cell cultures and host tissues was similar. This type of schizogony, previously undescribed in Eimeria, is compared with corresponding stages of development in other species of Eimeria and Sporozoa.     

毁灭泰泽球虫(Tyzzeria perniciosa)内生发育的超微结构研究——Ⅰ.裂殖生殖(Schizogony)

利用透射电镜对寄生于北京鸭小肠的毁灭泰泽球虫的裂殖生殖过程进行了观察。滋养体内未见多糖颗粒、脂肪体和致密体,在细胞质的被膜空泡内发现退化的微线和棒状体。在裂殖体核分裂过程中,出现典型的球虫型有丝分裂装置(如中心粒、中心锥、纺锤体)。裂殖子的发生是外瓣生方式,裂殖子在裂殖体的表面形成,并以母细胞的限制膜为外膜。     

Fine Structure of the Schizont and Merozoite of Isospora sp. (Sporozoa: Eimeriidae) Parasitic in Gehyra variegata (Dumeril and Bibron, 1836) (Reptilia: Gekkonidae)

SYNOPSIS. A study was made of the fine structure of some stages in the life cycle of an undesignated species of Isospora parasitic in a gecko. The merozoites which lay within a membrane-bound periparasitic vacuole in the host epithelial cell, had a striking similarity to Plasmodium, Lankesterella, Toxoplasma, Besnoitia, Sarcocystis, Eimeria and the M-organism. Each merozoite was invested with a triple-layered pellicle, the outer membrane of which was loosely applied. At the anterior end of the merozoite were conoid and apical rings; microtubules terminated in the posterior apical ring. Other organelles included nucleus, endoplasmic reticulum, mitochondria, micropyle, paired organelle, toxonemes and a variety of vacuoles. Although the sequence of development of the merozoite was not completely followed, some events in this process were recorded. The evidence suggests that anterior ends are formed early and that merozoites develop subsequently by a process of budding. The merozoite pellicle appears to be continuous with, altho structurally different from, the investing membrane of the parent cell.     

Fine Structure of the Endogenous Stages of Eimeria labbeana. I. The First Generation Merozoites

SYNOPSIS. The fine structure of the 1st generation merozoites of Eimeria labbeana from the ileal mucosa of artificially infected pigeons ( Columba livia ) was investigated and described. The 1st generation merozoites which appeared between 36-48 hr after infection averaged 4.4 × 2.1 μm in size. The 3-membraned pellicle was irregular in texture and harbored a single micropore, and many micropore-like invaginations. Closely apposed to the inner pellicular membrane were seen 22 microtubules, each 22–25 nm in diameter. An apical vesicle, 50 nm in diameter, seen at the anterior extremity, was connected with the common duct of the micronemes. The conoid consisted of 9 spiral elements, each 30 × 25 nm. The paired organelle (rhoptries) varied in length (1.4–2.2 μm), and the ductules (23 nm diameter) were composed of 2 inner tubules, each 6 nm in diameter. A unit membrane enveloped the partially alveolar and differentially osmiophilic interior of the bulbous regions of the rhoptries. The "rod-like structure"was found to be tubular and represented the common duct of the micronemes.     

The Fine Structure of Some Developmental Stages of Mattesia grandis McLaughlin (Sporozoa, Neogregarinida), a Parasite of the Boll Weevil Anthonomus grandis Boheman

SYNOPSIS Sporozoites, macronuclear schizonts, merozoites and gamonts of Mattesia grandis were examined by electron microscopy. A conoidal complex, consisting of conoid, polar rings and subpellicular microtubules was present in all of these stages. The conoidal complex was similar in structure to the same organelle of other Sporozoa. The conoidal complex in mono- to quadrinucleate macronuclear schizonts is transformed into an organelle similar to the mucron of some eugregarines.
This mucron consists of a specialized area of the cell membrane from which fine fibers extend into a large vacuole situated directly beneath the cell membrane. The top part of the vacuole is encircled by 2 ring-like structures formed by the dilatation of the original apical rings. The vacuole of the mucron contains many anastomosing protrusions of the cytoplasm, suggesting a nutritional role. The mucron disappears when the schizont reaches the multinucleate state. Later the merozoites bud from the surface of the schizont as in the coccidia. Each merozoite again has a conoidal complex, which persists thru the gamont stage and usually serves as the point of contact between 2 gamonts during their pairing.
The presence of a conoidal complex thru a major portion of the life cycle, its transformation into a mucron and the mode of formation of merozoites indicate that the Neogregarinida combine the fine structure characters of both the Eugregarinida and the Eucoccida, thereby suggesting a phylogenetic relationship between these sporozoans, with the neogregarines as a link between eugregarines and coccidia.     

La Genèse des Mérozoïtes chez la Coccidie Eimeria necatrix. Etude Ultrastructurale*

RESUME. Les schizontes de 2 ème génération d'Eimeria necatrix ont étéétudiés au microscope électronique. La différenciation des mérozoïtes est associée à la dernière mitose, qui ne semble pas différer essentiellement des précédentes. Les mérozoïtes se développent à la périphérie du schizonte. Le conoide et 22 microtubules sous pelliculaires, probablement induits par les centrioles, et le complexe membranaire interne ainsi que les précurseurs des rhoptries, qui semblent issus de l'appareil de Golgi, apparaissent auprès de chaque pôle nucléaire, sous la membrane du schizonte. Ces organites sont les premiers inclus dans les ébauches de mérozoïtes. Puis, le noyau, le dictyosome et les vésicules multimembranaires pénètrent dans les futurs mérozoïtes. Les micronèmes, probablement formés par l'appareil de Golgi, et les grains d'amylopectine sont produits plus tard, quand les mérozoïtes se séparent du reliquat cytoplasmique. Le mode de genèse de ces divers organites et les relations entre le dernière mitose et la différenciation sont discutés. SYNOPSIS. Second generation schizonts of Eimeria necatrix were studied with the aid of the electron microscope. Differentiation of daughter merozoites is associated with the last mitosis, which is not significantly different from the earlier ones. The merozoites develop at the periphery of the schizont. The conoid and 22 subpellicular microtubules, probably induced by centrioles, and the inner membranes complex and the rhoptry anlagen which seem to be produced by the Golgi apparatus, appear close to each nuclear pole, just near the schizont membrane. These organelles are the first to appear in the merozoite anlagen. Then, nucleus, dictyosome and multimembranous vesicles enter the budding merozoites. Micronemes, probably originating from Golgi apparatus, and amylopectin granules are produced later, when daughter merozoites separate from the residuum. The genesis of these various organelles and the relation between the last mitosis and differentiation are discussed.     

Ultrastructure of Sarcocystis tenella. I. Endozo?te (after negative staining]

The employment of negative staining technics for the endozoites (cyst stages) of Sarcocystis tenella allowed the elucidation of certain aspects of their fine structure. The conoid consists of similar to 20 oblique fibers and is surmounted by a ring with regular ornamentation. In the conoid's interior there are 2 excentric parallel microtubules which extend posteriorly for a considerable distance into the adjacent cytoplasm. The fibers of the conoid, intraconoid microtubules, appear to have the same diameter and structure as the 22 subpellicular microtubules. They are "cemented" anteriorly into a periconoidal ring which surrounds the conoid. The "reticulated" pellicle has certain differentiations: the micropore, surrounded by a "fibrillar" element, similar to 10 subcircular structures arranged into an anterior crown, and 11 rows of granules converging toward the posterior end. The sarconemes look like rice grains which, contrary to previous statements, are independent of one another. It is established that there are only 2 rhoptries.     

Fine Structure of Schizonts and Merozoites of Eimeria nieschulzi*

SYNOPSIS. Schizonts of E. nieschulzi lie in a vacuole within the host cell. After nuclear division the cell membrane invaginates forming merozoites. Differentiation of the pellicle and other organelles occurs while merozoites are still attached to the schizont cytoplasm. Merozoites have a pellicle thickened at the anterior end to form a polar ring. Radiating posteriorly from the ring, directly beneath the pellicle, are about 25 microtubules. Within the polar ring is a dense conoid. Extending posteriorly from within the conoid is a paired organelle. The paired organelle varies in size and shape in each generation of merozoites. Numerous toxonemes occupy the anterior half of the merozoites. Two paranuclear bodies are present in 1st generation merozoites. One or 2 granular bodies were seen in the anterior end of 2nd generation merozoites. In 3rd generation merozoites 6 or more granular bodies were seen anterior to the nucleus. Each merozoite has a single nucleus containing diffuse chromatin material. Elongate mitochondria and glycogen granules are present. The vacuole surrounding mature merozoites contains residual cytoplasm of the schizont and some granular material. Microvilli project into the vacuole from the host cell membrane.     

Ultrastructural study of developmental stages of Mattesia dispora (Neogregarinorida: Lipotrophidae), a parasite of the flour moth Ephestia kuehniella (Lepidoptera)

The ultrastructure of merozoites, gamonts and oocysts of the neogregarine Mattesia dispora and their development in larvae of the flour moth Ephestia kuehniella were studied by electron microscopy. The apical complex of free macronuclear merozoites was very distinct in micrographs of sections, the polar rings being especially prominent. Two gamonts associated in head-to-head syzygy and the apical complexes served as the contact point during pairing. At this stage the rhoptries became reduced and the conoid widened. The gamonts had a foam-like appearance in the light microscope. Paired gamonts formed an envelope and developed into a gametocyst, within which the gamonts were separated by a distinct border. Four gametes and two residual cells developed inside the gametocyst. The gametes were covered with a single membrane. The gametes fused in pairs to form two spherical zygotes, each covered by two membranes and with one large nucleus. The external layer appeared more undulated than the inner one. A single membrane covered each residual cell. Walls were formed around both zygotes to produce two oocysts. Each mature oocyst was lemon-shaped with polar plugs and eight peripheral sporozoites, which had a pellicle similar to that of the merozoites, lay beneath the thick oocyst wall.     

In vitro development from sporozoites to first-generation merozoites in Eimeria contorta Haberkorn, 1971. A fine structural study.

The asexual development of Eimeria contorta from sporozoites to first-generation merozoites in tissue culture was investigated with the electron microscope. Sporozoites with a three-layered pellicle, 26 subpellicular microtubules, a conoid, 4-7 rhoptries, and an abundance of micronemes actively entered host cells and showed direct contact to the host cell's cytoplasm. Shortly after penetration, small vacuoles surrounding the parasite merged into a parasitophorous vacuole. Inside this vacuole, sporozoites assumed a definite U-shape before transformation into schizonts took place. This process was characterised by the occurrence of subpellicular microtubules exclusively in the anterior half of the sporozoite, by a degeneration of the 2 inner pellicular membranes, by an outpocketing of the parasite's surface, and by the arrangement of microtubules in clusters. About 25 merozoites were formed at the surface of mature schizonts, to which they remained attached at their posterior pole. A polar ring was present at that area. Anterior and posterior refractile bodies were conspicuous in merozoites and showed close association with mitochondria. The significance of a fibrillar substructure in rhoptries and micronemes is discussed, and special attention is drawn to the pathway of nutrient transport from host cell mitochondria and dictyosomes through intravacuolar folds, parasitophorous vacuole and crescent body into the parasite's food vacuoles.     

Fine Structure of the Neogregarine Farinocystis tribolii Weiser, 1953. Developmental Stages in Merogony.

SYNOPSIS. The fine structure of schizonts and free merozoites of the neogregarine Farinocystis tribolii Weiser, and their development in the fat body of larval Tribolium castaneum were studied.
The surface of a multinucleate schizont and that of a uninucleate merozoite is covered by a double-layered membrane. Rhoptries and micronemes are present. The cytoplasm is packed with ribosomes and also contains dark bodies. Mitochondria are of the vesicular type. The spherical nucleus of the schizont and merozoite contains a large nucleolus. The anterior end of the merozoite has a typical conoidal complex composed of a conoid and a polar ring with 22 subpellicular mirotubules projecting from it.
New findings are a membranous septum across the body of the merozoite at 2/3 of its length below the nucleus and a highly osmiophilic spiral structure in the perinuclear space close to the Golgi complex. In addition, we found some "developmental stages" of the latter structure.     

Ultrastructural development of first- to second-generation merozoites in Eimeria contorta Haberkorn, 1971.

The development of first-generation merozoites to second-generation schizonts and merozoites of Eimeria contorta in one of its natural hosts, the mouse, was investigated with the electron microscope. Merozoites inside a host cell show a marked U-shape and a degeneration of the inner-pellicular membrane complex prior to transformation into schizonts. These processes closely resemble those seen in transforming sporozoites. In young schizonts with about 3-5 nuclei, the Golgi-adjuncts (structures of unknown function) form a large interconnected network. Nuclear divisions in growing schizonts involve the formation of a centroc?ne, which develops in a pocket-like indentation of the nuclear envelope. At least one centriole is present immediately adjacent to this indentation. In a later stage, the centroc?ne forms a conical nuclear protrusion directed towards a merozoite-anlage. This developing merozoite contains anlagen of a conoid, of rhoptries, and of micronemes and a refractile body in addition to the nucleus, centrioles, and a Golgi-adjunct. The merozoite-anlage is limited by a triple unit membrane complex. Schizonts give rise to 8-15 second-generation merozoites. Interesting features of these merozoites are the high number of micronemes, the finding of one single large mitochondrion per merozoite, and the occurrence of 26 subpellicular microtubules, i.e. the same number as in sporozoites of E. contorta. At the end of their development, merozoites come into direct contact with the host cell cytoplasm as the parasitophorous vacuole breaks down.     

Investigations into the fire structure of the asexual developmental stages of Frenkelia in the liver of the bank vole (author's transl)]

Bank voles (Clethrionomys glareolus) were infected by stomach tube with Frenkelia sporocysts from the faeces of buzzards (Buteo buteo). The voles were sacrificed at regular intervals and their livers examined electronmicroscopically. Seven days p.i. developmental stages of Frenkelia could be detected in liver parenchymal cells. The youngest schizonts detected are enveloped by a pellicle consisting of two membranes. This pellicle, which is in direct contact with the host cell mitochondria, shows marked invaginations which increase with the development of the schizont. A parasitophorous vacuole is not detectable. In developing schizonts numerous sections through nuclei with nucleic spindles and merozoite anlagen (dome-shaped) structures) are visible. It is not clear whether there are several nuclei or a section through one large and lobed nucleus. Within the merozoite anlagen the conoid and the subpellicular microtubules are formed first. By the prolongation of the dome-shaped structures towards the posterior pole, the nucleus and the other newly formed cell organelles are incorporated into the forming merozoite. The posterior pole of the merozoite still remains open at this stage of development. With increasing differentiation the merozoites become lancet-shaped, their apical poles bing always directed towards the periphery of the schizont. The outer membrane of the pellicle of the schizont forms the outer part of the pellicle of the merozoites by invaginating around them. At this stage of development the inner membrane of the pellicle of the schizont is no longer detectable. Thus the typical pellicle of the motile stages of sporozoaonsisting of three membranes is formed. In the centre of the merozoites which lie freely in the liver cell a residual body is present. The host cell reacts against the parasites by forming a thick border of mitochondria and distinct endoplasmic reticulum.     

Endogenous development of Eimeria intestinalis in rabbits (Oryctolagus cuniculus).

The endogenous life cycle of a pure strain of Eimeria intestinalis was studied by light and electron microscopy in coccidia-free rabbits. Four schizont generations could be observed: the first one, not previously described, was seen between 36 and 144 hr postinoculation (PI), the second one between 64 and 168 hr PI, the third one between 96 and 192 hr PI, and the fourth one between 168 and 240 hr PI. Gamogony apparently started as early as 144 hr PI. Thus, it was possible for oocysts to develop from third generation merozoites, later oocysts developing after the fourth schizont generation. Electron microscopic observation suggested that oocysts were derived mainly from merozoites of the fourth schizont generation. During the first stage of the life cycle, sporozoites were seen in intraepithelial lymphocytes. All asexual generations, except the fourth, were characterized by 2 schizont types: the first, regarded as female, contained mononuclear merozoites and the second, regarded as male, contained polynuclear merozoites.     

Fine structure of the merozoites of Barroussia schneideri parasitic in Lithobius forficatus

The ultrastructure of the merozoites of the parasite Barroussia schneiden (Bütschli, 1882) Reichenow & Schellack, 1912 in the intestinal cells of its centipede host, Lithobius forficatus (L) is described. The pellicle consists of a single outer and a double inner membrane under which there are 51 microtubules extending longitudinally. A micropore is present. The characteristic organelles and cytoplasmic inclusions of the merozoites of the Eimeriidac arc present: conoid, rhoptries (possibly 6), micronemes, nucleus with nueleolus, mitochondria with bulbous cristae, prominent Golgi complex, polysaccharide granules and granular endoplasmic reticulum.     

约氏疟原虫红外期成熟裂殖体的超微结构研究

用细胞色素氧化酶组织化学方法处理感染了约工疟原虫子孢子的大鼠肝脏,通过透射电镜研究红外期裂殖体的超微结构。在接种子孢子后４８小时的标本中发现一成熟裂殖体,外周仍由一寄生虫质膜包裹,膜下有许多小泡,粗面内质肉、圆形或蚕豆形具明显嵴的线粒体,以及大量成熟裂殖子。     

Fine Structure of the Endogenous Stages of Eimeria labbeana. 5. Schizonts and Merogony,with Special Reference to the Rhoptry-Microneme System*

SYNOPSIS The fine structure of the 3 generations of meronts, merogony, and merozoites of Eimeria labbeana Pinto from the ileal mucosa of experimentally infected pigeons, Columba livia Linnaeus, was described and compared to that of similar stages in other species of Eimeria. Sporozoite-trophozoite transition stages, trophozoites (5.8 × 4.2 μm), young meronts (10.1 × 8.4 μm), and mature meronts with free merozoites of the first generation, were observed at 20, 28, 36, and 48 hr post-infection, respectively. The 2nd and 3rd generation merogony were completed at 96 and 144 hr. Merogony was essentially of the ectomerogonous type without cytomere formation, as in most species. The average number of merozoites per meront in the 3 generations was 10 (5–15), 14 (8–19), and 7.5 (6–16); and the average size was 4.4 × 2.1 (4.1–5.9 × 1.8–2.2) μm, 4.2 × 1.8 (4.0–4.8 × 1.5–2.0) μm, and 5.4 × 1.8 (5.2–7.8 × 1.6–2.0) μm, respectively. Aggregation and subsequent degeneration of micronemes within membrane-bounded vesicles in the sporozoite-trophozoite stage, was observed as a possible mode of eliminating certain organelles present in the motile stages. Centrioles with (9 + 1) microtubular composition, and centrocones, were frequently seen in early meronts. Anlagen of micronemes, without any apparent association with the Golgi complex and the merozoite bud, were seen to develop in the cytoplasm of the meront. A single, median structure, probably representing the anlage of the rhoptry-microneme system was observed within the conoid of an early merozoite bud. Connections between the micronemes and the bulbous portion of the rhoptries, and a branched (interconnected ?) structure of the rhoptries observed in the present study, substantiate the present contention that the micronemes and rhoptries are functional forms of the same complex of organelles, the rhoptry-microneme system.     

Ultrastructural Observations on Renal Schizogony of Leucocytozoon dubreuili in the American Robin

SYNOPSIS. The schizogonic development of Leucocytozoon dubreuili in the kidney proximal tubule cells of the American robin, Turdus migratorius , was studied by electron microscopy. Renal schizogony is initiated by the entry of certain hepatic merozoites into cells of the proximal tubules. Development of the schizont consists of a coordinated sequence of events including extensive mitotic nuclear division, multiplication of mitochondria, increase in endoplasmic reticulum and ribosomes, differentiation of membranes, microtubules, micronemes and rhoptries, and cytoplasmic segmentation (cytomere formation). Merozoites form by budding around numerous centers in the schizont and, when mature, are bounded by a single plasma membrane subtended by microtubules. Each merozoite contains a large nucleus, a mitochondrion, and a well developed apical complex consisting of 3 polar rings, paired rhoptries, and numerous micronemes.
An atypical nuclear division observed in some maturing schizonts was characterized by the multiple fission of a nucleus within a persistent outer nuclear membrane and the absence of mitotic spindle apparatus. Alterations in infected renal cells consisted of disorganization and loss of cytoplasmic organelles and the accumulation of lipofuscin-like inclusions.     

Fine structure of human malaria in vitro.

The erythrocytic cycle of the human malaria parasite, Plasmodium, falciparum, was examined by electron microscopy. Three strains of parasites maintained in continuous culture in human erythrocytes were compared with in vivo infections in Aotus monkeys. The ultrastructure of P. falciparum is not altered by continuous cultivation in vitro. Mitochondria contain DNA-like filaments and some cristae at all stages of the erythrocytic life cycle. The Golgi apparatus is prominent at the schizont stage and may be involved in the formation of rhoptries. In culture, knob-like protrusions first appear on the surface of trophozoite-infected erythrocytes. The time of appearance of knobs on cells in vitro correlates with the life cycle stage of parasites which are sequestered from the peripheral circulation in vivo. Knob material of older parasites coalesces and forms extensions from the erythrocyte surface. Some of this material is sloughed from the host cell surface. The parasitophorous vacuole membrane breaks down in erythrocytes containing mature merozoites both in vitro and in vivo. Merozoite structure is similar to that of P. knowlesi. The immature gametocytes in culture have no knobs.     

Ultrastructure of the pellicular complex of Plasmodium fallax

The exoerythrocytic merozoites of Plasmodium fallax grown in a tissue-culture system have been investigated by negative staining and thin-sectioning techniques, and the respective results have been compared. Negative staining provided additional information, corroborated findings obtained with thin sectioning, and contributed particularly to the study of the pellicular complex of the merozoites which has been demonstrated as being composed of three layers: a thin outer membrane, a thick interrupted inner membrane, and a partial layer of microtubules. Observations made of negatively stained parasites revealed that the thick, interrupted inner membrane in thin sections is actually a labyrinthine structure and covers the entire surface of the merozoite, except at the regions of the conoid and the cytostome. The microtubules which radiate from the conoid to the posterior end demonstrated a transverse periodicity and filamental subunits parallel to the axis of the microtubule. The detailed structure of the conoid and the cytostome is also described.