Basal body replication and ciliogenesis in Tokophrya infusionum were studied in synchronized cultures. Basal body replication occurs during the 1st hr of reproduction, which in Tokophrya is by internal budding. The number of basal bodies increases from about 20 to over 300 within this period. New basal bodies develop in association with mature basal bodies; they are formed at right angles to the mature basal body as short "probasal" bodies, which elongate, slant upward, become parallel to the mature basal body, and elongate to the mature size. Ciliogenesis occurs only during reproduction; the nonreproducing adult is not ciliated, and has only 18–25 barren basal bodies. Cilia first appear as short bulges above the basal body. The axonemal structure is incomplete at first, with one or both central microtubules absent, and occasionally the B fibers of the outer doublets are missing. Several accessory fibers are associated with the basal bodies, both in the adult and during reproduction. One of the fibers appears only after the cilia have sprouted. The scheme of basal body replication and ciliogenesis in Tokophrya is compared to that reported in other organisms, and the role of the accessory fibers is discussed.

The feeding apparatus of Suctoria consists of long, thin, stiff tubes called tentacles. When a swimming prey attaches to the tip of the tentacle a number of events follow in rapid succession. The tentacle broadens, a stream of tiny granules starts to move upward at its periphery to the tip, the prey becomes immobilized and shortly thereafter the cytoplasm of the still living prey begins to flow through the center of the tentacle to the body of the predator. An electron microscope study of the tentacle in Tokophrya infusionum , a protozoan of the subclass Suctoria, has disclosed a number of structural details which help to clarify some of the mechanisms involved in this unusual way of feeding. Each tentacle is composed of two concentric tubes. The lumen of the inner tube is surrounded by 49 tubular fibrils most probably of contractile nature. In the inner tube the cytoplasm of the prey is present during feeding, and in the outer tube are small dense bodies. It was found that the dense bodies originate in the cytoplasm of Tokophrya . They have an elongate, missile-like appearance, pointed at one end, rounded at the other, and are composed of several distinct segments. At the tip of the tentacle they penetrate the plasma membrane, with their pointed ends sticking out. It is assumed that the missile-like bodies play a major role in the feeding process. Their composite structure suggests that they might contain a number of enzymes which most probably are responsible for the various events preceding the actual food intake.

Blood collected from rats infected with Plasmodium berghei was centrifuged and the pellet was fixed for 1 hour in 1 per cent buffered OsO 4 with 4.9 per cent sucrose. The material was embedded in n -butyl methacrylate and the resulting blocks sectioned for electron microscopy. The parasites were found to contain, in almost all sections, oval bodies of the same density and structure as the host cytoplasm. Continuity between these bodies and the host cytoplasm was found in a number of electron micrographs, showing that the bodies are formed by invagination of the double plasma membrane of the parasite. In this way the host cell is incorporated by phagotrophy into food vacuoles within the parasite. Hematin, the residue of hemoglobin digestion, was never observed inside the food vacuole but in small vesicles lying around it and sometimes connected with it. The vesicles are pinched off from the food vacuole proper and are the site of hemoglobin digestion. The active double limiting membrane is responsible not only for the formation of food vacuoles but also for the presence of two new structures. One is composed of two to six concentric double wavy membranes originating from the plasma membrane. Since no typical mitochondria were found in P. berghei , it is assumed that the concentric structure performs mitochondrial functions. The other structure appears as a sausage-shaped vacuole surrounded by two membranes of the same thickness, density, and spacing as the limiting membrane of the body. The cytoplasm of the parasite is rich in vesicles of endoplasmic reticulum and Palade's small particles. Its nucleus is of low density and encased in a double membrane. The host cells (reticulocytes) have mitochondria with numerous cristae mitochondriales. In many infected and intact reticulocytes ferritin was found in vacuoles, mitochondria, canaliculi, or scattered in the cytoplasm.

Contractile vacuoles are organelles that collect fluid from the cytoplasm and expel it to the outside. After each discharge (systole), they appear again and expand (diastole). They are widely distributed among Protozoa, and have been found also in some fresh water algae, sponges, and recently in some blood cells of the frog, guinea pig, and man. In spite of the extensive work on the contractile vacuole, very little is known concerning its mode of operation. An electron microscope study of a suctorian Tokophrya infusionum provided an opportunity to study thin sections of contractile vacuoles, and in these some structures were found which could be part of a mechanism for the systolic and diastolic motions the organelle displays. In Tokophrya , as in Suctoria and Ciliata in general, the contractile vacuole has a permanent canal connecting it with the outside. The canal appears to have a very elaborate structure and is composed of three parts: (1) a pore; (2) a channel; and (3) a narrow tubule located in a papilla protruding into the cavity of the contractile vacuole. Whereas the pore and channel have fixed dimensions and are permanently widely open, the tubule has a changeable diameter. At diastole it is so narrow (about 25 to 30 mµ in diameter) that it could be regarded as closed, while at systole it is widely open. It is assumed that the change in diameter is due to the contraction of numerous fine fibrils (about 180 A thick) which are radially disposed around the canal in form of a truncated cone, with its tip at the channel, and its base at the vacuolar membrane. It seems most probable that the broadening of the tubule results in discharge of the content of the contractile vacuole. In the vicinity of the very thin limiting vacuolar membrane, small vesicles and canaliculi of the endoplasmic reticulum, very small dense particles, and mitochondria may be found. In addition, rows of closely packed vesicles are present in this region, and in other parts of the cytoplasm. It is suggested that they might represent dictyosome-like bodies, responsible for withdrawing fluids from the cytoplasm and then conveying them to the contractile vacuole, contributing to its expansion at diastole.

In a previous paper (8) an organized structure was described in the macronuclei of certain old organisms of Tokophrya infusionum . It was found that the same honeycomb structure appears in great abundance in the macronuclei of overfed organisms. This permitted a better three-dimensional reconstruction of the described structure. Since the defined structure may be experimentally induced, it offers an opportunity for further more detailed studies as to its nature and meaning.

A study of thin sections of Paramecium multimicronucleatum, Tetrahymena pyriformis, Tokophrya infusionum , and Amoeba proteus shows that the mitochondria in all these protozoa are similar in certain aspects of their fine structure to that described in metazoan cells. As in higher organisms the mitochondrion is surrounded by a double limiting membrane and contains protrusions directed inward from the innermost of the double membranes. There are, however, some differences. In a majority of higher organisms the internal structure of mitochondria consists of ridges or cristae mitochondriales and in a few instances only of finger-like projections, or microvilli. In all protozoa described here and elsewhere microvilli represent the dominant structure. They are characteristic therefore of protozoan mitochondria.

The macronucleus in Tokophrya infusionum is composed of numerous Feulgen-positive chromatin bodies (about 0.5 µ in diameter) which appear in thin sections as a dense spongework, homogeneous throughout. The same appearance characterizes metaphase chromosomes of higher forms. Some chromatin bodies of the macronucleus were found to possess a highly organized structure in certain old organisms. This structure appears in cross-sections as a honeycomb and in longitudinal sections as parallel lines about 120 A in diameter evenly spaced (about 230 A). As far as is known this is the first time a regular structure has been found in bodies of chromosomal character at the dimensional level presently explored by electron microscopy. The demonstration that OsO 4 can preserve order in chromatin material is another significant aspect of these findings.

OsO 4 has many advantages over Carnoy's fixative mixture for the Feulgen nuclear staining in the protozoan Tokophrya infusionum . While Carnoy's fluid used prior to the Feulgen reaction produces shrinkage of the macronucleus and coarse clumping of its chromatin bodies, OsO 4 preserves faithfully the size and shape of the macronucleus and its chromatin material. This finding seems to be of special importance in view of the fact that electron microscopy relies on OsO 4 fixation. The satisfactory preservation of structured detail in Feulgen-stained preparations is of importance for the correlation of histochemical and morphological information.