Preparation and Chemical Composition of the Cell Membranes of Developmental Reticulate Forms of Meningopneumonitis Organisms

The outer limiting membranes of developmental reticulate forms of the meningopneumonitis organism were purified by a combination of differential centrifugation, trypsin digestion, and sodium dodecyl sulfate treatment, and their physical and chemical properties were compared with those of outer envelopes of mature dense forms of this organism. Reticulate bodies were easily disrupted by short periods of sonic treatment and were lysed by trysin digestion, in contrast to the dense bodies which were resistant to these treatments. In electron micrographs, reticulate body membranes were seen as very thin, flattened structures, whereas dense-body envelopes showed folding rigid membranes. The results of chemical fractionation of (32)P-labeled purified preparations indicated that reticulate body membranes have smaller amounts of phospholipid, and are more dense than cell walls of the mature forms. The analysis of amino acid composition of reticulate body cell membranes showed that they do not contain cystine or methionine, both of which were found in cell walls of dense bodies. These results clearly show that there are significant differences in the chemical and physical properties of the outer envelopes of the developmental and mature forms of this organism.     

Studies on the cytophysiology of the fat body of the American silkmoth

Summary A developmental study at the electron microscopic level was conducted of the fat body cells of Hyalophora cecropia (L.). During the last larval instar the fat body increases in volume and the cells exhibit a well developed rough endoplasmic reticulum and protein bodies of diverse sizes. In the pupal fat body, the protein bodies appear to be enclosed by a double membrane and contain glycogen granules, ribosomes and mitochondrion-like structures. In addition, there are large lipid globules, cytolysomes and rough endoplasmic reticulum. The ultrastructure of the protein bodies suggests the development of large bodies by fusion of smaller protein bodies. Changes in fat body cell ultrastructure were followed during adult development and cytological evidence was obtained for the depletion of protein, glycogen and lipid in the female during this period. The female adult fat body cell contains free ribosomes, protein bodies, many mitochondria, a few lipid globules and glycogen granules. The male moth fat body cells have many mitochondria, a few glycogen granules, essentially no protein bodies, but an abundance of large lipid globules.Studies on the influence of egg maturation on the morphology of the fat body of Hyalophora gloveri (L.) revealed that ovariectomy of pupae yielded adults having more fat body than normal females, and that the fat body cells of the ovariectomized animals contained more glycogen, lipid and protein. Male pupae receiving ovarian implants developed into adults containing eggs and possessed more fat body than normal females but less than normal males. Very few glycogen granules were found in the fat body cells of normal males or males with implanted ovaries.Supported by grant AM-02818 from the National Institutes of Health.We thank Dr. James Oschman for his helpful suggestions and constructive criticisms.     

Cytomembranes in first cleavage xenopus embryos

Summary The ultrastructure and interrelationships of the Golgi body, endoplasmic reticulum and lipid droplets have been studied in the first cleavage Xenopus embryos. Lipid droplets, usually spherical or sometimes multilobed, did not have a discernible limiting membrane, although some had an incomplete electron dense partition. The Golgi bodies and endoplasmic reticulum were seen continuous with lipid droplets and the profiles indicated a probable formation of these membranes from lipid droplet material. Rough endoplasmic reticulum (ER) mainly consisted of paired tubular cisternae and vesicles containing filamentous material that gave a fringed appearance. The relationships of paired cisternae with the Golgi body suggested a transformation of ER membranes into the Golgi body membranes. In addition, paired ER cisternae showed a close apposition with the limiting membrane of the yolk platelet. Lone ER cisternae that contained moderately electron dense material instead of filaments were also present and showed numerous associated vesicles near the Golgi body. The Golgi body showed several morphological forms including a single fenestrated cisterna, two to four flat or cup-shaped cisternae, or up to seven cisternae, some of which were dilated and similar to fringed ER in appearance. These forms could be different developmental stages of the organelle. Coated vesicles were seen continuous with the cisternae of the Golgi body. A probable route for the assembly of the cell surface material has been proposed.This work was supported by a grant from the Medical Research Council of Canada to one of us (E.J.S.).     

Fine structural study of a possible mechanism of secretion by the interrenal cells of the brown pelican

Summary In the deeper zones of the adrenal gland of the brown pelican, small dense bodies have been observed in the subendothelial space and between adjacent interrenal cells. They appear to be extruded from the interrenal cell by reverse pinocytosis. Intracellularly, similar appearing dense bodies, bounded by a single membrane, are sometimes evident. An interpretation of the foregoing is presented and discussed which suggests that this material may be hormone which is synthesized or organized into droplet form by the agranular endoplasmic reticulum and extruded from the cell either apically or apico-laterally, to be dispersed or rendered soluble in the subendothelial space.Supported in part by the Comly Fund of the Ohio State University, by Grant AM 09658-01 from the National Institute of Arthritis and Metabolic Diseases, National Institutes of Health and Grant DT-8 from the National Institute of Dental Research.Deceased January, 1965.     

Fine Structural Observations of the Erythrocytic Stages of Plasmodium chabaudi Landau, 1965*

SYNOPSIS. Electron microscopic examination of Plasmodium chabaudi in mouse erythrocytes revealed many characteristics resembling those observed in other mammalian malarial parasites. A double unit membrane surrounds the trophozoite cytoplasm which contains many ribonucleoprotein particles, a limited amount of endoplasmic reticulum and membraned organelles including sausage-shaped vacuoles and multilaminated membraned bodies. More or less circular double membraned vacuoles, possibly cross sections of the sausage-shaped vacuoles, are common. Typical protozoan mitochondria are lacking. The limiting membrane of the merozoites is triple-layered. Paired organelles and small dense bodies are found in the merozoites along with dense granular masses in the nuclei. Trophozoites have cytostomal structures as well as invaginations of the plasma membrane at sites where no cytostomes are evident. Digestion appears to occur in single membrane-bound vesicles which contain one to several pigment grains. P. chabaudi frequently contains multiple food vacuoles and has polymorphism manifested in part by the presence of cytoplasmic extensions and of nuclei with a variety of shapes. Several apparently free forms are noted, often accompanied by a thin rim of host cytoplasm. “Appliqué” forms are common among the trophozoites as are forms in which 2 or more trophozoites are joined together. Finally, alterations in the host cytoplasm resembling the socalled Maurer's clefts are frequent. Ferritin-containing vacuoles also appear in the host cell.     

Formation of yeast mitochondria III: Biochemical properties of mitochondria isolated from a cytoplasmic petite mutant

A number of biochemical properties of mitochondria from a cytoplasmic petite mutant ofSaccharomyces cerevisiae with an extremely high adenine plus thymine content have been studied.When such particles are isolated by means of standard procedures developed for use with wild-type yeasts they are grossly contaminated by non-mitochondrial membrane fragments. Further enrichment of mitochondria is achieved by non-equilibrium centrifugation in sucrose gradients.Throughout this purification procedure the particles can be shown to retain an outer limiting, as well as a non-cristate inner membrane. In many of their morphological and physical features (size, shape, buoyant density) they resemble mitochondria isolated from the wild type.Although enzymes of the respiratory chain are absent from the mutant particles, their content ofl-malate dehydrogenase, NADP-dependent isocitrate dehydrogenase, and ATPase is comparable to that found in the wild type. The mitochondrial ATPase in this mutant strain is cold stable, oligomycin insensitive, Dio-9 sensitive, and susceptible to inhibition by the F₁ inhibitor of beef heart. The enzyme can be rendered cold labile by its detachment from the membrane, followed by fractionation with protamine sulfate and ammonium sulfate.The existence of mutant particles that are incapable of function in oxidation and phosphorylation but resemble their functional homologues in many other ways raises the possibility that mitochondria are required in the cellular economy for purposes not directly linked to oxidative phosphorylation and electron transport. This hypothesis has led us to suggest that, contrary to models currently under discussion, mitochondria did not evolve as a consequence of endosymbiosis. We propose as an alternative that the mitochondrial organelle evolved as a means of improvement of existing subcellular structures in the primordial (perhaps eukaryotic) cell. Partial autonomy may thus constitute a relatively recent modification; the present-day mitochondrial genome had its origin in nuclear DNA and may have been amplified in a manner not unlike the amplification of ribosomal RNA cistrons in developing oocytes ofXenopus.Supported by Research Grant GM 12228 from the National Institute of General Medical Science, National Institutes of Health, U.S. Public Health Service.Recipient of a Public Health Service Career Award No. GM 05060 from the Institute of General Medical Sciences.     

Dense bodies of the contractile system of cardiac muscle in Venus mercenaria

Dense bodies in the heart muscle of Venus mercenaria exist in two forms, free and attached. Free dense bodies morphologically consist of fascicles of thin filaments in parallel array and bound together by a dense, amorphous proteinaceous material. The binding of dense bodies to the cell membrane is effected via connecting filaments of the amorphous material of the dense body which join a condensation of morphologically similar material attached to the inner osmiophilic layer of the unit membrane. This composite of dense body, connecting filaments, membrane condensation and unit cell membrane has been termed collectively the attachment plaque. The attachment plaque is part of an extensive network on the cell surface which obligates that surface to a role in the contractile process. Moreover, this set of attachment plaques imposes an organization and an orientation to most thin filaments of the cell and preserves the contractile axis of the cell.     

A study of the ultrastructure of the shoot apex and leaf cells in two liverworts,with special reference to the oil bodies

Summary In this investigation attention has been paid to the general ultrastructure of the shoot apical and leaf cells in the liverwortsBazzania trilobata andLophozia ventricosa but especially to the different developmental stages of their oil bodies. These species have been chosen because their oil bodies differ from each other in size and shape.The appearance of the different organelles, nucleus, chloroplasts, mitochondria, ER, and Golgi bodies, are in their main features the same as those of higher plants described in the literature. The dark cytoplasm seen in the leaf cells ofLophozia in the vicinity of the oil bodies but without any surrounding membrane when fixed in double fixative 2, seems to be specific to this species. On the other hand, granular dense bodies were visible in the cells of the shoot apex ofBazzania, which shrank in size as the development of the oil bodies proceeded and were lacking in the mature leaf cells.In both species investigated, the oil bodies have the same component parts: (1) an outer membrane enveloping the whole body, (2) inside this, a granular stroma layer of varying thickness enveloping (3) specific globules of varying size and number, each of which is surrounded by (4) a thin inner membrane (Fig. 28).The oil bodies develop in at least two ways and usually in one way for each species. InBazzania they seem to develop from vacuole-like formations in the shoot apex or in the leaf primordia into which substances have segregated. InLophozia they seem to originate by aggregation and fusion of lipid bodies.     

Nuclear and cytoplasmic differentiation in developing sperm of the crayfish,Cambaroides japonicus

Summary In the present electron microscopic study of spermatogenesis in the crayfish, Cambaroides japonicus, it was possible to clarify several aspects of the unusual differentiation which leads to the production of an aflagellate sperm. The centriole is followed from the metaphase of the second spermatocyte division to the time at which, in the nearly mature sperm, it appears to disintegrate. It has no connection with the acrosome but in the late spermatid and maturing sperm it is found randomly oriented among the convoluted membranes of the filamentous endoplasmic reticulum.There appears to be a close association of mitochondria with the developing acrosomal vesicle. Typical mitochondria, however, are not present after the late spermatid stage of development. It is suggested that the complex lamellar bodies associated with the nuclear envelope in the late stages of spermatogenesis may be related to mitochondria for these lamellar bodies resemble the complex mitochondria found in the adjacent nutritive cells.The development of the acrosome has been traced from an aggregate of dense granules which first appear in the interzonal spindle region and are later segregated at one side of the cell after the second spermatocyte division. As differentiation proceeds, tubular elements appear and disappear within the acrosome, while somewhat later, fibrous elements appear in the matrix. In the mature acrosome, the fibrous elements remain only adjacent to the granular periphery of the acrosome and the core again becomes homogeneous.No typical Golgi complex is found in these cells at any time during their differentiation.In the maturing sperm the development of the arms of the nucleus was studied. Preceding the differentiation of the arms a coarse fibrous material develops in the periphery of the nucleus. It is shown that the fibrillar material in the matrix of the arms is in continuity with the fibrillar material in the matrix of the nucleus proper.Supported in part by Grant No. B 2314 of the National Institute of Neurological Diseases and Blindness, U.S. Public Health Service.Predoctoral Research Fellow of the National Institute of Neurological Diseases and Blindness, U.S. Public Health Service.     

Sperm tail differentiation in the nudibranch mollusc Hypselodoris tricolor (gastropoda,opisthobranchia)

The sperm axoneme of Hypselodoris tricolor forms from a single centriole that is located initially beneath the plasma membrane and then migrates to the nuclear surface. A conspicuous centriolar adjunct-like formation is present in the neck of midspermatids, but it becomes very reduced at the end of spermiogenesis. In spermatocyte and spermatid mitochondria, intracristal bodies originate from the accumulation of a dense material in some cristae. From our observations and foregoing reports, it may be concluded that the process of sperm tail differentiation in opisthobranchs resembles that in pulmonates, whereas it differs in many respects from that occurring in prosobranchs. The appearance of intracristal bodies in modified mitochondria seems to be a special feature of spermatogenesis in the opisthobranchs that does not occur in the two other groups of gastropod molluscs.     

AN ELECTRON MICROSCOPE STUDY OF THE EPITHELIUM IN THE NORMAL MATURE AND IMMATURE MOUSE CORNEA

1. An electron microscope study at high resolution of the corneal epithelium of the normal mature and immature mouse revealed new information regarding the submicroscopic appearance of these cells. 2. Two thin dense lines separated by a less dense area constituted the structure of the limiting surface membrane of epithelial cells; the thickness of this membrane was about 80 A. 3. Some differences in the appearance of the cytoplasm and mitochondria of cells from the immature mouse cornea and the appearance of the cytoplasm and mitochondria of cells from the adult mouse cornea were observed. 4. The basement membrane appeared as a dense band about 600 A wide separating the basal epithelial cells from the substantia propria. Suggestions of periodicity were seen in some phosphotungstic acid-treated specimens. 5. Round bodies believed to be bacteria were seen on the surface of the outer epithelial cells in the adult mouse cornea but not in the immature, unopened eye.     

Comparative studies on the histology of the ovotestis in Hypselodoris tricolor and Godiva banyulensis (Gastropoda,Opisthobranchia), with special reference to yolk formation

The histology of the ovotestis was studied by light and electron microscopy in two nudibranch gastropod species. While in Hypselodoris tricolor the ovotestis is intimately associated with the digestive gland tissue, the large gonadal mass of Godiva banyulensis is placed freely in the haemocoele. This fact results in great histological differences between both species. As is common among Mollusca, the immature yolk granule in Hypselodoris and Godiva presumably originates from membrane-rich cytoplasmic inclusions, which we have termed dense multivesicular bodies. Such inclusions consist of an outer membrane enclosing membrane remnants and a granular, electron-dense material. These elements are accumulated and mixed in the center of the dense multivesicular body and could be actually transformed into the paracrystalline core of the immature yolk granule, the cortex of which is made up of part of the central accumulation materials that have not spread into the crystal. During vitellogenesis, some mitochondria are subjected to a process of transformation affecting mainly their inner membrane (including mitochondrial cristae) and matrix. However, the conversion of modified mitochondria into yolk precursors, as reported for other gastropod species, could not be determined with absolute certainty on the basis of our observations on static material. The mature yolk granule consists of a central paracrystalline core, similar in structure to that of the immature yolk granule, and a peripheral membranous cortex, which seems to spread centripetally, thus permitting the crystal to grow. The cortical material consumed in synthesizing the central core appears to be restored by addition of degenerative mitochondria to the yolk granule surface.     

Balbiani bodies in cricket oocytes: Development, ultrastructure, and presence of localized RNAs

Formation of two spherical Balbiani bodies along the long axis of previtellogenic oocytes in Acheta domesticus was demonstrated by differential interference microscopy. The structures form adjacent to and on opposite sides of the germinal vesicle, the anterior body first. Each migrates to the nearest pole of the elongating oocyte and retains its spherical structure until occluded from view by accumulating yolk. In situ hybridization, immunocytochemistry, and confocal immunofluorescent microscopy showed Balbiani body components to include y-tubulin, alpha-tubulin, EF1alpha, and several RNAs homologous to localized Xenopus RNAs implicated in embryonic axis formation or germ cell determination. The latter include Xcat2, Xwnt11, Xlsirt, and Xpat. Balbiani body ultrastructure includes a dense cloud of tubular mitochondria, rough ER, Golgi-like membrane aggregates, and microtubules. The results suggest that molecules and mechanisms specifying early determinative events for embryogenesis in vertebrates and insects are highly conserved and that Balbiani bodies may have a role in establishing developmental asymmetry in the cricket.     

The reticular substance of the medulla oblongata of the albino rat

Summary The region of the reticular giganto-cellular nucleus, perfused with formalin and postfixed in osmium tetroxide, was studied with histochemical and electron microscopic techniques. The perikarya of the neurons have two zones. The peripheral cytoplasm contains Nissl bodies, mitochondria, and free RNP particles. The juxtanuclear cytoplasm contains the Golgi complex, mitochondria, RNP particles and dense bodies. The nucleus is indented and has a prominent nucleolus and a paranucleolar body. Dense bodies are found along the axon and dendrites as well. Three different types of synapses are described and two types of synaptic vesicles (spherical and ellipsoidal) are shown.The capillary endothelium shows microvilli and marginal flaps. The endothelial cytoplasm contains vacuoles, micropinocytotic vesicles, and a few dense bodies. Processes of pericapillary cells, surrounded by a basement membrane, also contain dense bodies. The dense bodies found in the neurons and endothelial cells show acid phosphatase activity. On the basis of their morphology and their enzymatic activity these bodies are identified as lysosomes.Partially supported by a school grant No RF 62051 from the Rockefeller Foundation, New York, USA, and Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina.Fellows of the Consejo National de Investigaciones Científicas y Técnicas, Argentina. The authors wish to thank Dr. Mario H. Burgos for his constant interest and criticism, and to Dr. Eduardo Rodriguez Echandia and Dr. Fabio L. Sacerdote for their valuable assistance.     

中国大鲵卵母细胞发育的显微和超微结构

用光镜和电镜观察了大鲵卵母细胞在发育过程中的显微和超微结构变化，着重对类核周体结构和线粒体与卵黄前颗粒的关系进行了详尽观察。贴近卵核的类核周体由核仁样体和线粒体群构成，远离卵核的类核周体仅由线粒体群构成。线粒体群是线粒增殖区，其中有多种形态的原线粒体，有些处于增殖状态，它们未形成明显的线粒体嵴。散在于卵质中的线粒体是成的线料体，有明显的嵴，其中许多线粒体内沉积着致密物质，一些致密物质从线粒体中向外     

In vitro maturation of Arbacia punctulata oocytes and initiation of heavy body formation

Summary An ultrastructural study of in vitro maturation of A. punctulata oocytes was undertaken to determine when heavy body formation was initiated. No heavy bodies were seen in germinal vesicle oocytes or in oocytes undergoing germinal vesicle breakdown or polar body formation. Heavy bodies were only observed in ova examined one to two hours after pronuclear formation. Several small heavy bodies were seen in sections of eggs fixed as early as three hours after the pronucleus had formed. The number of these structures in the egg cytoplasm increased with time. Therefore it is concluded that heavy body formation in sea urchin ova is a phenomenon following nuclear maturation.Contribution No. 222 from the Institute for Molecular and Cellular Evolution. Supported in part by grants from the National Institutes of Health (5-T01-HD00026-09 to the Fertilization and Gamete Physiology Training Program at the Marine Biological Laboratory and predoctoral fellowship 1-F01-GM-36,719-01A1 to C. M. Conway) and the National Science Foundation (GB3899 to C. B. Metz).The authors are grateful to Drs. A. F. Conway, Giovanni Giudice, and Gertrude W. Hinsch for consultation and criticism of this work.     

Ultrastructure of the hypodermis during cuticle formation in the third molt of the nematode Nippostrongylus brasiliensis

Summary The fine structure of the hypodermis of Nippostrongylus brasiliensis, a parasitic nematode of the rat, was studied in free-living third-stage filariform (infective) larvae, and in parasitic forms undergoing the third molt. In filariform larvae the hypodermis displayed a comparatively poorly developed RER, and few Golgi complexes, mitochondria, and vesicles. Nuclei contained large amounts of heterochromatin and no nucleoli. After worms reached the lungs and the third molt began, the RER, Golgi complexes, mitochondria, multivesicular bodies, and coated vesicles greatly increased in amounts. Nuclei displayed less heterochromatin and contained prominent nucleoli. These morphological changes were associated with transition from the free-living to the parasitic mode in life. The results were correlated with the fact that the hypodermis in N. brasiliensis is actively synthesizing culticular collagen during the third molt.This investigation was supported, in part, by awards 1-F02-AI3750-02 and AI-09625 from the National Institutes of Health, United States Public Health Service.We are grateful to Lida Petruniak and Dora Lou for technical assistance.     

Vesicle and granule content of sympathetic ganglion cells during limb regeneration of the newt Triturus

Summary Fine structural observations were made on the vesicle and granule content of ganglion cells in the posterior subclavian ganglion and peripheral nerve fibers of the upper forelimb of the newt Triturus. The populations of vesicles and granules in normal ganglion cells and nerve fibers were compared with those observed after limb transection. In normal neurons, clear vesicles range in size from 250 to 1000 Å in diameter, but are most frequently 400–500 Å. Vesicles with dense contents (granules) also vary greatly in size, but most are 450–550 Å in diameter and correspond to dense-core vesicles. Large granules that contain acid phosphatase activity are thought to be lysosomes. During limb regeneration, in both the ganglion cells and peripheral nerves, the ratio of dense vesicles to clear vesicles increases. There is a large increase in number of dense granules with a diameter over 800 Å, particularly in the peripheral regenerating fibers. This study shows that regenerating neurons differ from normal in their content of vesicular structures, especially large, membrane-bounded granules.This work was supported by grants from the National Science Foundation (GB 7912) and from the National Cancer Institute (TICA-5055), National Institutes of Health, United States Public Health Service.     

Herring bodies; an electron microscopic study of local degeneration and regeneration of neurosecretory axons

Summary The Herring bodies in the posterior lobe of the bovine hypophysis are very large (2–600 ) and can be classified into three types. The type I Herring body contains an accumulation of neurosecretory granules. These Herring bodies are very scarce and should not be confused with the numerous, but small, axonal swellings which also contain neurosecretory granules.The type II Herring body is characterized by the presence of a varying number of normal, moderately electron dense and empty vesicles, autophagic vacuoles, multilamellate bodies and occasional mitochondria. These Herring bodies are frequently observed.The type III Herring body is typified by the presence of dense vesicles connected to tubular formations which contain material of variable electron density, of filaments, and of long slender and very numerous mitochondria.The presence of multilamellate bodies and autophagic vacuoles suggests that the type II Herring body is in a degenerating phase. This concept is further substantiated by the similarity between this type of Herring body and transected neurosecretory axons in which degeneration is occurring.A similar comparison suggests that the type III Herring body is undergoing a regenerative process. Our current concept of the structure and function of Herring bodies is revised in the discussion.This work was supported by grants 5 RO1 NB 06641 NEUA and 5 R0107492 NEUA from the National Institutes of Health and the Space Sciences Research Center of the University of Missouri. The technical assistance of Mrs. G. Clark and Mr. R. Faup, and the clerical assistance of Mrs. S. Schmidt are gratefully acknowledged.Fellow of the Conséjo National de Investigaciones Científicas y Tecnicas de la República Argentína.     

Light and electron microscopy on the sporulation of the oocysts of Eimeria brunetti. II. Development into the sporocyst and formation of the sporozoite

The later stages of sporulation in oocysts of Eimeria brunetti were examined in samples which had been allowed to sporulate at 27 degrees C for 24, 36 and 48 hours. It was observed that the sporoblasts became ellipsoidal and the nucleus underwent the final division. A nucleus with associated Golgi bodies was not observed at either end of the organism. The cytoplasm was limited by two unit membranes and contained rough endoplasmic reticulum, dense bodies, electron translucent vacuoles and mitochondria. The first evidence of sporozoite formation was the appearance of a dense plaque at either end of the organism. This appeared in the vicinity of the nuclei, and adjacent to the limiting membrane of the soroblast. At this stage the sporocyst wall was still unformed. Then the two sporozoites were formed from opposite ends of the organism by growth of the dense plaques and invaginations of the plasmalemma which thus formed the pellicles of the developing sporozoites. A conoid and subpellicular microtubules were observed at this stage as development continued, a number of vacuoles were found between the nucleus and the conoid. These vacuoles constituted the precursors of the rhoptries and micronemes. At the same stage a large dense body had appeared within the forming sporozoite. As the sporozoite developed, this body, anterior refractile body, is followed by the nucleus and another dense body which formed the posterior refractile body. During this period, the thin sporocyst wall was formed and Stieda and sub-Stieda bodies were now present at one end of the sporocyst. Each mature sporocyst contained two sporozoites.