Showing papers in "Journal of Eukaryotic Microbiology in 1967"

Taxonomic Criteria for Limax Amoebae, with Descriptions of 3 New Species of Hartmannella and 3 of Vahlkampfia*

Abstract: SYNOPSIS. Seven species of limax amoebae were isolated into clonal, monoxenic cultures with Aerobacter aerogenes from material collected from freshwater habitats. Studies were made of their trophic structure, nuclear division, cyst structure, some aspects of cytochemistry, and other characteristics. Six new species are described: Vahlkampfia inornata, V. avara, V. jugosa, Hartmannella limacoides, H. vermiformis, and H. exundans. The well-known species Naegleria gruberi (Schardinger, 1899) is re-described on the basis of 8 strains; its flagellated phase was found to be biflagellate, with rare exceptions. A correlation exists between the manner of locomotion and the pattern of nuclear division in the limax amoebae in the family Vahlkampfiidae and those in the genus Hartmannella. Trophic amoebae of all species had a PAS-positive surface layer, altho results with H. vermiformis and H. exundans were less definite than with other species. All species except H. limacoides formed cysts in culture. The cyst walls of all cyst-forming species were strongly PAS-positive, but results of the zinc chloroiodide test for cellulose were negative with the method used. The genus Hartmannella Alexeieff, 1912, is re-defined to include those species which assume a simple, monopodial limax-like form during locomotion and have nuclear division similar to that of metazoan cells and to distinguish it from the genus Acanthamoeba Volkonsky, 1931.

Fine structure, reconstruction and possible functions of components of the cortex of Tetrahymena pyriformis.

Abstract: SYNOPSIS. Improved methods of preparing cells for electron microscopic study have permitted a more complete examination of the tubular and fibrous elements already reported in the cortex of Tetrahymena pyriformis. Three of these structures, a kinetodesmal fiber, a band of postciliary microtubules and a band of transverse microtubules, are all associated at one of their ends with the proximal end of a basal body-cilium complex. From this position they radiate out as tho from one corner of a tetrahedron, each passing separately to one of the other 3 corners. Available evidence is presented and discussed for a structural function for these elements. A firm bonding of these fibers and bands to the basal body at one of their ends and to the amorphous material under the pellicle at their other ends is thought to provide for this support. Connecting the proximal ends of the left side of the basal bodies of a kinety is another, previously undescribed, set of tubules. Their diameter, 24 mμ, and cross sectional structure are similar to those of the other microtubules. However, their more sinuous longitudinal appearance, small number of tubules per kinety and different reaction to fixatives suggest a different function. Because of their location it is proposed that they may be a communication line between basal body-cilium complexes of a kinety. A 3-dimensional drawing shows the positioning of the above structures in the cortex. Bodies with an internal tubular structure appear anterior to the proximal end of some basal bodies. They are referred to as probasal bodies due to their resemblance to procentrioles; they may be immature basal bodies. Smooth-membraned cisternae which have bristle-coated pits and bear a resemblance to the outer pellicular membrane appear in the cytoplasm. Their origin is discussed.

Re-definition of the genus Acanthamoeba with descriptions of three species.

Abstract: SYNOPSIS. Ten strains of Acanthamoeba from freshwater habitats were isolated in clonal cultures. Studies were made of trophic structure, nuclear division, cyst structure, some aspects of cytochemistry, and other characteristics. One strain was identified as A. castellanii (Douglas, 1930), one as A. astronyxis (Ray and Hayes, 1954), and 8 as A. polyphaga (Puschkarew, 1913). Strains of Acanthamoeba isolated by other workers were also examined comparatively. The pattern of nuclear division in all strains resembled that in metazoan cells, with the exception that centrioles were never found. Trophic amoebae had a PAS-positive surface outline. Cyst walls were strongly PAS-positive and also gave a positive test for cellulose with zinc chloroiodide. The genus Acanthamoeba Volkonsky, 1931 is re-defined, being distinguished from Hartmannella Alexeieff, 1912, emend. Volkonsky, chiefly by the formation of tapering, hyaline pseudopods (acanthopodia) and by a cyst made up of an ectocyst and a polyhedral or stellate endocyst, with excystment by removal of opercula. Other characteristics found in all strains include a distinctive food cup, the presence of many small refractile globules in the cytoplasm of trophic amoebae, and a cyst wall containing cellulose. The degree of spindle convergence, employed by Volkonsky as a generic criterion, was unusable. Differential diagnoses based principally on cyst structure are offered for A. castellanii, A. astronyxis, and A. polyphaga. The strain previously called Mayorella palestinensis Reich, 1933 is a distinct species of Acanthamoeba.

Differentiation of Trypanosoma cruzi in culture.

Abstract: SYNOPSIS. A medium containing sodium and potassium chloride, phosphate buffer, glucose, calf serum, hemoglobin and lactalbumin hydrolysate permits some growth and differentiation of crithidiae into the metacyclic forms of Trypanosoma cruzi. The addition of ox liver infusion greatly enhances the growth promoting properties of this medium while making it very poor as far as differentiation is concerned. With dog heart infusion instead of ox liver, the medium is still good for growth but not much better for differentiation. If, however, the pH of the dog heart infusion medium is lowered from 7.2 to 6.7, it continues to be good for growth and becomes excellent for differentiation. Other factors were studied that stimulate or repress the rate of differentiation, such as preincubation at certain levels of temperature and the size and age of inocula.

Electron Microscopic and Histochemical Studies of Sporozoite Formation in Plasmodium berghei

Abstract: SYNOPSIS. Observations were made on the differentiation of fine structure during sporogonic development of Plasmodium berghei. The oocyst in the process of sporozoite formation is an encapsulated structure 30-40 μ in diameter. It typically develops while in an extracellular position, attached to the basement membrane of the mosquito midgut and projecting into the mosquito hemocoel. Occasionally, however, ookinetes passing thru the midgut epithelial cells may become impacted within a cell so that the resulting oocyst develops intracellularly. Each oocyst has a large differentiating region, the sporoblastoid body. This body contains large dividing nuclei which are Feulgenpositive, and a cytoplasm which includes mitochondria, dense rodlike structures, cytoplasmic membranes, cisternae and vacuolar structures, Golgi material, and ribosomes which are both free and membrane-associated. Sporozoite budding takes place along the surface of the sporoblastoid body. Bits of a new membrane condense under the plasma membrane which bounds the sporoblastoid body. These 2-membraned sites then bulge out, continue to elongate, and eventually become sporozoites. The various nuclear and cytoplasmic components of the sporoblastoid body are passed into the sporozoites during their elongation. In addition, the sporozoite develops a system of elogate, subpellicular microtubules, possibly contractile in function. The pellicle of the sporozoite is broken by an opening, the cytostome (micropyle). The anterior end is truncate.

Development of first-generation schizonts of Eimeria bovis in cultured bovine cells.

Abstract: SYNOPSIS. Monolayer primary and secondary cultures of embryonic bovine kidney, spleen, intestinal and testicle cells, and secondary cultures of embryonic bovine thymus, maintained in lactalbumin hydrolysate, Earle's balanced salt solution and ovine serum were observed for a maximum of 21 days after inoculation of E. bovis sporozoites. The sporozoites entered the cells in all of these cultures but underwent development only in primary cultures of kidney and intestinal cells and in secondary cultures of kidney, spleen, thymus, intestinal, and testicle cells. In acellular media, the sporozoites retained motility no longer than 21 hr. In the cell cultures, free motile sporozoites were seen for as long as 18 days after inoculation. Sporozoites entered cells anterior end first; the process of penetration required a few seconds to about a minute. Sporozoites were also observed leaving host cells. Intracellular sporozoites were first seen 3 min after inoculation; they were observed at various intervals up to 18 days after inoculation. In transformation of sporozoites into trophozoites a marked change in size and appearance of the nucleus took place before the change in shape of the body occurred. Trophozoites were first found 7 days after inoculation, multinucleate schizonts after 8 days, and schizonts with merozoites after 14 days. Schizonts containing merozoites were seen only in kidney, spleen, and thymus cells. The mature schizonts were smaller and represented a much lower proportion of the total number than in comparable stages of infections in calves. Schizonts with many nuclei occurred in intestinal cells; the most advanced stage seen in testicle cells was the binucleate schizont. Nuclear and cytoplasmic changes were observed in the infected cells.

Trichodina (Ciliata: Urceolariidae) of freshwater fishes of the Southeastern United States.

Abstract: SYNOPSIS. Eight new species of Trichodina are described from freshwater fishes of the Southeastern U.S.: T. davisi from Roccus saxatilus (Walbaum), T. funduli from Fundulus notti (Agassiz), T. globosa from Etheostoma radiosum (Hubbs and Black), T. hoffmani from Etheostoma edwini (Hubbs and Cannon), T. hypsilepis from Notropis hypsilepis Suttkus and Raney, T. microdenticula from Dorosoma petenense (Gunther). T. noturi from Noturus leptacanthus Jordan, and T. salmincola from Salmo gairdneri Richardson. Nine species of Trichodina are redescribed: T. californica, T. discoidea, T. fultoni, T. pediculus, T. platyformis, T. reticulata, T. sp. (?T. nigra), T. tumefaciens, and T. vallata. The name T. pediculus is re-erected for the form that occurs on largemouth bass. Host fish were collected from 36 geographic locations in 10 states (Alabama, Arkansas, Florida, Georgia, Kentucky, Louisiana, Mississippi, North Carolina, South Carolina, and Tennessee). A total of 939 fish of 46 species was examined for trichodinid parasites. Two methods of impregnating formalin preserved specimens of Trichodina with silver are discussed. A key to the species of Trichodina reported from North American freshwater fishes is given.

Ultrastructural Observations on the Development of the Microsporidian Protozoon Plistophora hyphessobryconis Schaperclaus

Abstract: SYNOPSIS. A sequence of developmental stages of Plistophora hyphessobryconis Schaperclaus, a microsporidian protozoan parasite of the muscular tissue of several species of freshwater fishes, was studied with the electron microscope. The youngest stages observed, ca. 4 × 2 μ, have a single nucleus and their plasm contains only ergastoplasmic lamellae and ribosomes. They are surrounded by a halo of lysed host tissue. They increase in volume to become large sporonts with a great number of nuclei and a thick, 2-layered membrane. Thru schizogony, a corresponding number of sporoblasts is produced within this pansporoblast membrane. Sporoblasts start to develop a thick spore membrane, and a number of smooth-membraned vesicles appear in the plasm. These vesicles fuse to make the outer membrane of the filament. Later, its inner structures originate—the axial electron-dense substance, filling the hollow lumen of the filament, and a middle, electron-transparent layer. The structure of the filament is discussed in relation to its function and with regard to the findings of other authors. The polaroplast is a laminated structure, originating possibly by transformation of endoplasmic reticulum; the polar cap forms its apical part. The cap is also lamellar; its substance reaches into the lumen of the filament for a certain distance. No micropyle was discovered in the shell; the filament is fastened to the polar cap. These observations on microsporidian development and on the structure of their spores are compared with similar data on myxosporidian species. Such a comparison speaks clearly in favor of the complete taxonomic separation of the Microsporidea from the Myxosporidea, the latter being quite different also from other sporozoa sensu lato.

Crithidia mellificae n. sp. an acidophilic trypanosomatid of the honey bee Apis mellifera.

Abstract: SYNOPSIS. Crithidia mellificae n. sp. is described from the honey bee Apis mellifera in apiaries of Victoria, Australia. Organisms were isolated and cultured in a modified NNN medium with Cowper-thwaite's medium as an overlay. In contrast to other isolates of Crithidia, C. mellificae grows best at a pH between 4.5 and 5.5 with optimum near pH 5. In Cowperthwaite's medium alone, buffered at pH 4.5–5.5, the organism will survive apparently indefinite passages. The organism has so far as known, no pathologic effects in the hymenopteran host.

Some Blood Sporozoans from East African Reptiles

Abstract: SYNOPSIS. Six hundred and forty-one lizards belonging to 28 species, and 29 snakes belonging to 12 species from East Africa were examined for blood sporozoans. Eight species of lizards and 4 of snakes were positive. Tissue phases were seen in some infections; no proven arthropod vectors were found. Parasites are described from previously unreported hosts; new host records, and new stages in the life histories of previously named parasites are reported. The sporozoans belong to the genera Plasmodium, Haemoproteus, Haemogregarina or Hepatozoon, and probably Pirhemocyton. Reasons are given for the inadvisability of establishing new species of some of these on the basis of blood stages alone.

Ultrastructure of the Giant Amoeba Pelomyxa palustris

Abstract: SYNOPSIS. The ultrastructure of the herbivorous amoeba Pelomyxapalustris was studied. Nuclear division is not understood in this amoeba, and evidence for the method of nuclear division was sought. This species typically has many spheroidal nuclei which are similar within a given cell. However, some amoebae from our collections differed from this common type in both the number and structure of their nuclei. This suggested stages associated with nuclear division. One current hypothesis of nuclear division in this organism is that of nuclear budding. Our evidence is more in accord with this method than with mitosis. The cytoplasm contained no mitochondria, Golgi bodies, contractile vacuoles or crystals. Most amoebae had 2 types of bacteria (bacteroids or endosymbionts) in their cytoplasm; a separate vesicle enclosed each of these. Characteristically, only 1 type of bacterium (Bn) surrounded the nucleus. Another type (B) was found elsewhere in the cytoplasm. Also in the cytoplasm were the following: food vacuoles enclosing various algae, relatively clear vacuoles and vesicles, glycogen, various electron-opaque particles, and occasional microtubules. The plasmalemma was smooth, lacking the external fringe which characterizes other large fresh-water amoebae.

The Mastigont System of Trichomonas gallinae (Rivolta) as Revealed by Electron Microscopy

Abstract: SYNOPSIS. The fine structure of Trichomonas gallinae has been examined by electron microscopy and correlated with previous light microscope observations. A composite diagram of the flagellate, derived from both types of examination, is presented. Details of relationships of various mastigont organelles are documented by electron micrographs. The extent of the pelta and its connection to the capitulum of the axostyle have been determined. Four types of kinetosome rootlets have been described. One consists of superficial “filaments” radiating from each of the 9 triplet microtubules of kinetosomes #1, #2 and #3. A 2nd type of rootlet structure is represented by single comma-shaped filaments emerging clockwise from kinetosomes #1 and #3. The filament from kinetosome #1 has a periodic structure similar to that of the marginal lamella with which it is believed to connect. A 3rd type of rootlet emerges from kinetosome #2 as a sheet of about 9 filaments which traverse a sigmoid course and terminate on the inner surface of the microtubules of the pelta near the peltar-axostylar junction. The 4th set of structures consists of the costa and parabasal filaments. These structures have major periodicities of similar dimension but have readily differentiable repeating units. The costa appears to originate at the kinetosome of the recurrent flagellum, but its origin is also contiguous with that of parabasal filament 2 which has some continuity with kinetosomes #2 and #3. Parabasal filament 1, on the other hand, arises solely from or near kinetosome #2. Occasional observations of a costa and a parabasal filament in juxtaposition over a great part of their length has led to the suggestion that the parabasal filament may play a role in the development of the costa. Periodic and filamentous structures have been observed in paraxostylar and paracostal granules and in nearby cytoplasm. Their possible role in providing substance for the developing axostyle and the costa is discussed. The results are discussed in the light of available information pertaining to structure of various trichomonad species as revealed by light and electron microscopy.

The Process of Chamber Formation in the Foraminifer Rosalina floridana (Cushman)

Abstract: SYNOPSIS. The foraminifer Rosalina floridana builds a chambered, calcareous test which is periodically enlarged by the addition of new chambers. R. floridana begins to form a chamber by constructing an algal growth cyst which covers the dorsal side of the animal and is cemented to the substrate. The pseudopods that build the cyst coalesce to form a cytoplasmic template or anlage on which the chamber walls will be secreted. Electron micrographs reveal the anlage cytoplasm to be a “froth” of nearly empty vesicles which contain mitochondria, fibrillar material, and electron dense granules. The organic lining of the new chamber is secreted on the anlage by pseudopods extending thru it. After the organic lining is completed, cytoplasm from within the test flows into the forming chamber and forces the anlage cytoplasm out thru the new aperture. This “frothy” anlage cytoplasm forms a sheath over the dorsal surface of the test; while it is in place, a layer of calcite is deposited on the walls of the new chamber and over the rest of the test. When calcification is completed, the sheath breaks up and is incorporated into newly formed pseudopods as the foraminifer gradually moves out of the growth cyst and begins normal feeding. The production of vesiculated cytoplasm in normal pseudopods and in the anlage is viewed as a method of greatly increasing cytoplasmic volume with a resultant very small loss in cytoplasmic mass. In the anlage during the production of the organic lining the vesiculated cytoplasm apparently acts only as a support for the membranes being secreted by the pseudopods and presumably does not take part in the secretory process. This same cytoplasm forms the sheath that is present during calcification. The view is advanced that, altho the sheath could be active during calcification with the mitochondria within the vesicles actively transporting calcium to sites of crystal growth, its more probable function is to form a partition between the parts to be calcified and the environment. The foraminifer could then secrete CaCO3 from pooled reserves in the cytoplasm into the area between the sheath and the chamber membranes. The basal membrane is considered the nucleating agent during calcification and is responsible for the ordering of the crystals so their C axes are perpendicular to the surface of the chamber.

Ultrastructure of Pellicular and Ciliary Structures of Euplotes eurystomus

Abstract: SYNOPSIS. Many of the sub-pellicular and infraciliary structures in protozoa have proved difficult to study with standard thin-sectioning technics. When these structures are viewed in isolated and fragmented form, many of the thin-sectioning difficulties are circumvented. Langmuir-trough isolation followed by critical-point drying, as well as thin sectioning, were used in this study to determine the patterns of sub-pellicular microtubules and fibrils interconnecting kinetosomes of membranelles and cirri of Euplotes eurystomus. The fibrillar network in the bases of these ciliary organelles is presented in some detail and apparent variations in pattern are noted. Functional aspects of some of the structures are discussed. With special preparation nearly whole Euplotes may be obtained for study in the electron microscope. Fused cilia were frequently obtained and their ultrastructure was studied.

Microstome-macrostome transformation in Tetrahymena vorax strain V2 type S induced by a transforming principle, stomatin.

Abstract: SYNOPSIS. Microstome macrostome transformation in Tetrahymena vorax was induced by suspending microstomes in a transforming principle, stomatin, released by a potential prey, T. pyriformis. It was found that 70–90% of the microstomes formed macrostomes within 7 hours following suspension in this transforming principle. Macrostome formation occurred by the process of oral replacement. This process involved resorption of the microstome oral apparatus and its replacement with a larger (macrostome) one, which arose from an anarchic field that formed behind the resorbing oral area. Ninety-five percent of those microstomes which were destined to form macrostomes were in some stage of oral replacement 195 minutes after their suspension in stomatin. Several commercially produced products were tested over a wide range of concentrations to determine their ability to act as an inducer of macrostomes. Only 2, Trypticase and Bactocasitone, had any activity, and it was too small to be considered really effective. An attempt was also made to destroy the activity of stomatin by using enzymes. RNAse was effective but only in very high concentrations, so it was suggested that this activity might be related to the destruction of RNA within the transforming cell and not related to hydrolysis of stomatin. None of the other enzymes tested had any effect in reducing the activity of stomatin.

Fine Structure of Microgametocytes and Macrogametes of Eimeria nieschulzi

Abstract: SYNOPSIS. An electron microscope study of microgametocytes and macrogametes of Eimeria nieschulzi Dieben, 1924 revealed that they lie within vacuoles bounded by a host unit membrane. The vacuole surrounding the microgametocyte contains granular material. The vacuole around the macrogamete is narrower and contains vesicles and membranes. Micropores were seen on the surface of the plasma membrane of microgametocytes and macrogametes. Microtubules were seen in macrogametes. Young microgametocytes and macrogametes have a similar cytoplasmic matrix, mitochondria and nuclei. Glycogen granules apparently develop around vacuoles in both microgametocytes and macrogametes. Glycogen granules were also seen along the margins of parallel bundles of fibers in microgametocytes. As nuclei of the microgametocyte divide, they move to the periphery of the parasite. Three basal bodies, each with 9 fibers in triplet form, develop in association with each nucleus. Microgametes have 2 free flagella and a central short, attached flagellum. Basal granules lie along the outer fibers of the central flagellum. Each microgamete has an elongate mitochondrion in close contact with the nucleus. In macrogametes wall-forming bodies develop in lacunae in the cytoplasm. Smaller dark bodies with areas of low density were also seen. Wall-forming bodies and dark bodies move to the periphery of mature macrogametes.

A Proposed Life Cycle of Minchinia nelsoni (Haplosporida, Haplosporidiidae) in the American Oyster Crassostrea virginica

Abstract: SYNOPSIS. A developmental sequence is proposed for the haplosporidan Minchinia nelsoni Haskin, Stauber and Mackin, 1966, based on study of oyster infections over the past 5 years in Chesapeake Bay. Uninucleate stages develop by nuclear division into multinucleate plasmodia which proliferate in the tissues by plasmotomy. Relatively small plasmodia containing what are considered to be gametic nuclei originate by unequal plasmotomy of large plasmodia. These have been interpreted to aggregate and fuse to form large plasmodia which contain prozygotes. Pairing and fusion of nuclei occur within each plasmodium to produce zygote nuclei (synkaryons) which undergo division, possibly meiotic, to form sporonts. Sporoblasts differentiate into spores with the development of spore walls and opercula. Cystoid plasmodia develop during times of unfavorable conditions. An anomalous but common sequence involving sexuality and mitosis is described, and the occurrence of various life cycle stages within the host thruout the year is discussed.

Schizogony and Gametogony of Leucocytozoon simondi and Associated Reactions in the Avian Host

Abstract: SYNOPSIS. Stages of development of Leucocytozoon simondi in White Pekin ducklings and their reactions to the parasite were studied on successive days after infecting them artificially with sporozoites from Simulium rugglesi. The minimum prepatent period was 5 days. The first asexual cycle occurred exclusively in the parenchymal cells of the liver. Progeny of these hepatic schizonts followed one of 3 courses: (a) invaded parenchymal liver cells to give rise to another hepatic cycle, (b) penetrated blood cells to form round gametocytes, and (c) were phagocytized by macrophages and grew into megaloschizonts thruout the body. The appearance of elongating gametocytes coincided with the period of maturation and release of merozoites from the megaloschizonts. Experimental evidence supports the hypothesis that the round gametocytes arise from the hepatic schizonts and the elongate forms from the megaloschizonts. Mature megaloschizonts released millions of merozoites, but a high 2nd peak in parasitemia did not develop because of retention of developing gametocytes in the deep circulation, particularly the liver and spleen, and a pronounced host reaction.

Infection of macrophages in culture by leptomonads of Leishmania donovani.

Abstract: SYNOPSIS. Peritoneal macrophages from hamsters were monolayered on coverslips in Leighton tubes. Twenty-four hours later these were transferred to a perfusion chamber. Leptomonads were added with fresh medium and the infection process observed with the aid of phase contrast. In the perfusion chamber free-swimming leptomonads attached to the macrophage by the tip of their flagella. Shortly after this initial attachment the macrophage extended a narrow pseudopodium around the flagellum which eventually reached and enveloped the body of the parasite. Upon complete envelopment the pseudopod containing the leptomonad was retracted into the central body of the macrophage. When first seen in the granular endoplasm of the macrophage, most of the leptomonads appeared to be surrounded by vacuoles. In most cases these vacuoles disappeared in a few minutes making it difficult to distinguish the parasite from the host cell cytoplasm. Leptomonads also were added directly to Leighton tube cultures, and the coverslips with the adherent macrophages and parasites were removed, fixed and stained periodically during the infection process. In these preparations most of the parasites were in clumps in the vicinity of macrophages. Details of the ingestion of the clumps could not be seen, but occasionally a single organism was seen with its flagellum and part of its body enclosed by an extended pseudopod. Most of the intracellular leptomonads were in large vacuoles. Forms intermediate between elongate leptomonads and LD bodies were surrounded by smaller vacuole-like spaces. The halo-like vacuoles most frequently seen around LD bodies may have been fixation artifacts. Under favorable conditions the leptomonads transformed to LD bodies in 1–4 hours, but it was 48 hours before a population increase could be found.

Characterization and Localization of Acid Phosphatase Activity of Trypanosoma gambiense

Abstract: SYNOPSIS. Properties and cellular location of acid phosphatase in Trypanosoma gambiense were studied. Activity was found in both the sediment (32,000 ×g) and the supernatant of homogenates. Cenrifugation in 0.3 M sucrose showed activity principally in the lowspeed fraction (4,000 ×g). One min of sonication released most of this activity. Several phosphomonoesters were hydrolyzed at acid H's. Enzymatic activity was relatively specific for pyrophosphate and p-nitrophenylphosphate at pH 3.6. At pH 5.2, purine and pyimidine nucleotide 5′-triphosphates as well as adenosine di- and ono-5′-phosphates were hydrolyzed nonspecifically. Activity with yrophosphate at pH 3.6 had a temperature optimum of 60-70 C while that for adenosine 5′-triphosphate (pH 5.2) was 50 C. These ctivities of the sediment required no metal co-factors and were inibited by Fe++, inhibition at the lower pH being greater. Glucose 6-phosphate was hydrolyzed by the supernatant with maximum activity between pH 6.0 and 7.2 and a temperature optimum of 50 C. This pH range showed a broad plateau with 2 or 3 minor peaks. The hydrolysis of p-nitrophenylphosphate showed a similar pH curve. In glucose 6-phosphate hydrolysis, Mg++ was a required co-factor but could be replaced by Ni++ or Co++. Ammonium sulfate fractionation precipitated most of the supernatant activity between 50 and 75% saturation. A modified Gomori technic produced spherical deposits of PbS thruout the cytoplasm of the intact cell. With the electron microscope, Pb phosphate deposition was observed in membrane-bound vesicles (i.e., lysosomes) approximately 100-150 mμ in diameter. These organelles were common in the region of the reservoir at the base of the flagellum. Acid phosphatase activity specific for glucose 6-phosphate as substrate was localized within this basal pocket.

The ultrastructure of Crithidia fasciculata and morphological changes induced by growth in acriflavin.

Abstract: SYNOPSIS. Crithidia fasciculata is similar to other trypanosomatids in ultrastructure. Of considerable interest is the finding of a lamellar membrane formation attached to the mitochondrion. Some evidence is provided for the tentative hypothesis that this membrane formation is the precursor of mitochondrial membranes. The cellular origin of this structure is unknown. Growth in acriflavin results in a marked alteration of the mitochondrion and kinetoplast. Both structures are deficient in cristae. In addition, the kinetoplast loses its normal fibrillar appearance and becomes a smaller, more electron-dense organelle. These effects are discussed in relation to the proposed involvement of the kinetoplast in the elaboration of functional mitochondria.

Ultrastructure of the Eyespot and its Possible Significance in Phototaxis of Tetracystis excentrica

Abstract: SYNOPSIS. The eyespot of the zoospore of Tetracystis excentrica (a green alga) has been studied by light and electron microscopy. In Tetracystis the eyespot consists of about 110 osmiophilic granules which form a plate in the anterior third of the cell. The granules are about 80 A in diameter and are found in the outermost portion of the chloroplast; they commonly show hexagonal close packing and a hexagonal shape. The granules are confined positionally by the chloroplast envelope and an inner thylakoid. The plasmalemma over the eyespot is thickened and is separated from the chloroplast envelope by a 50 mμ space. The eyespot of Tetracystis is compared with others reported in the literature and the possible functional significance of these studies is discussed. The possibility that the eyespot plate in Tetracystis serves as a shading device rather than the primary photoreceptor is considered.

Cochlodinium heterolobatum n.sp.: Structure and Some Cytophysiological Aspects*

Abstract: SYNOPSIS. Structure and morphogenesis, and cytochemical data on Cochlodinium heterolobatum, a new species of unarmored dinoflagellate, were derived from living and fixed material from culture. C. heterolobatum is characterized by the torsion of the girdle which descends in a left-hand spiral 1.8 turns; the sulcus having a torsion of 0.8 turn; a sulcus loop in the epicone; a tongue-shaped lobe in the right hypocone; nucleus in the epicone; and a stigma in the left epicone. Trichocysts and behavior of the nucleus during typical and atypical divisions are described in cells from cultures of different ages. A small form with the specific characters was found. Intracellular bacteria were seen and their growth followed in individuals from cultures of different ages. A possible relationship between those bacteria and the accumulation of metabolites inside old cells is discussed.

The Test Structure and Composition of the Foraminifer Rosalina floridana

Abstract: SYNOPSIS. The composition of the test of Rosalina floridana (Cushman) was examined histochemically, and its structure was studied with the electron microscope by means of thin sections and carbon replicas. The test is composed of a thick organic lining overlain by one or more calcite layers bounded above and below by thin membranes. The membranes are fused to organic pore processes composed of coarse fibers that penetrate the calcite layers. The ***lining, consisting of coarse fibers matted into a laminated sheet, is considered a strengthening element of the test. The membranes covering each calcite layer are composed of fine, headed fibrils which in aggregate have a striated pattern; they are thought to be the crystal-nucleating agent during calcification and to form a protective covering for the previously deposited calcite layers. The pore processes, which are devoid of an internal entrance for cytoplasm, are considered to be points of attachment for the membranes; they tie the organic test components into a unified whole. The calcite layers and the chambers lack this unity, being separated from each other and from the preceding chambers by membranes so that there are no calcite-to-calcite boundaries between them. An organic, sievelike structure of undetermined function has been found in the foramina of chambers near the prolocular region of the test. Histochemical methods show that the lining contains proteins, polysaccharides, and unidentified substances; the membranes and the pore processes stain as a protein-polysaccharide complex free of other substances.

A comparative study of mitosis in amoebae and plasmodia of the true slime mold Didymium nigripes.

Abstract: SYNOPSIS. Studies comparing mitosis in amoebae and plasmodia of the true slime mold Didymium nigripes reveal that at the time of differentiation pronounced changes occur in the mitotic process. Not only does the amount of time required for division of the 2 stages differ, but plasmodial mitosis is characterized by persistence of the nuclear membrane and the apparent lack of centrioles. The origin of multinucleate plasmodia from uninucleate cells which have already undergone cytoplasmic differentiation is described. Division time in a population of amoebae becomes more uniform after those cells which are destined to form plasmodia have differentiated. The observations and data presented indicated that differences in mitotic behavior also occur between amoebae of 3 stocks with differences in plasmodial structure and behavior. Comparison of mitosis in the plasmodia of these 3 stocks revealed no significant differences.

The Glyoxylate Pathway in Acanthamoeba Sp.

Abstract: SYNOPSIS. Cell-free extracts of encysting Acanthamoeba were assayed for the key enzymes of the glyoxylate pathway, viz., isocitrate lyase and malate synthase. Both enzymes were present at the onset of encystment but their activities changed as cyst-wall formation proceeded to completion. Isocitrate lyase activity decreased during the first 4 hr of encystment to a minimum at 4 hr which was 70% of its initial activity. Activity then increased reaching a maximum at 9 hr which was 144% of its initial activity. After 9 hr a decrease in isocitrate lyase activity began which reached 70% of its initial activity at 35 hr. Malate synthase activity slowly decreased throughout encystment to 50% of its initial activity after 35 hr. From these data and others cited, it is concluded that this small soil amoeba has a functional glyoxylate pathway.

Comparative cultivation of poultry coccidia in mammalian kidney cell cultures.

Abstract: SYNOPSIS. Excysted sporozoites of Eimeria meleagrimitis, E. necatrix, E. acervulina, and E. gallopavonis were inoculated into monolayer cell cultures of bovine, ovine, porcine, and human kidney. E. meleagrimitis developed only in bovine embryonic kidney. Mature schizonts were found in the 11th, 16th, and 20th serial passages, but only immature schizonts were in the 4th and 6th passages. E. necatrix developed to mature schizonts in the 3rd, 4th, 6th, 11th, 16th, and 20th passages of bovine kidney and also to immature schizonts in the 175th and 189th passages of PK-15 (cell line porcine kidney). Schizonts, however, did not develop in the 140th and 145th passages of CCI-33 (cloned PK-15). Neither E. meleagrimitis nor E. necatrix developed in the primary, 1st or 2nd passages of bovine embryonic kidney, primary porcine kidney, 45th and 52nd passages of a human embryonic kidney cell line, or in the primary, 5th and 18th passages of ovine kidney. Eimeria acervulina and E. gallopavonis did not develop in any of the cultures. E. meleagrimitis and E. necatrix probably completed only one asexual generation in culture. The structure of mature schizonts of both species differed greatly from those in the natural host. Schizonts of E. meleagrimitis present at 48 hours were small (13–18 by 12–14 μ) and contained only 12–28 merozoites that were 3.2–3.8 μ long. At 48 hours, E. necatrix schizonts were 15–18 μ in diameter or less and contained only 15–20 merozoites (2.0–3.5 μ long); at 96 hours they were 50–70 by 10–35μ and contained either hundreds of small merozoites (2.0–3.5 μ long) or a lesser number of larger merozoites (9–11 μ).

Coccidia (Protozoa: Eimeriidae) of the Alpaca Lama pacos*

Abstract: SYNOPSIS. Three new species of coccidia are described from the alpaca Lama pacos from Peru. These are the first species of coccidia to be named from this host. The oocysts of Eimeria lamae n. sp. are ellipsoidal, occasionally ovoid, 30–40 by 21–30 μ (mean 35.6 by 24.5 μ) with elongate ovoid sporocysts 13–16 by 8–10 μ (mean 15.3 by 8.5 μ). The oocysts of Eimeria alpacae n. sp. are ellipsoidal, rarely ovoid, 22–26 by 18–21 μ (mean 24.1 by 19.6 μ), with ovoid sporocysts 10–13 by 7–8 μ (mean 11.0 by 6.8 μ). The oocysts of Eimeria punoensis n. sp. are ellipsoidal, occasionally ovoid, 17–22 by 14–18 μ (mean 19.9 by 16.4 μ), with ovoid sporocysts 8–11 by 5–7 μ (mean 9.2 by 6.1 μ).

The Effect of a Eugregarine Gregarina polymorpha (Hammerschmidt) on the Mealworm Larva of Tenebrio molitor (L.)

Abstract: SYNOPSIS. The larva of the beetle Tenebrio molitor (Coleoptera, Tenebrionidae), commonly referred to as the mealworm, harbors considerable numbers of gregarines in its midgut. These are not necessary for normal growth, nor do they prolong the life of larvae grown under optimal conditions of temperature, relative humidity and diet. When the larvae were grown under optimal conditions there was no significant difference between the length of larval life or the final pupal weight of mealworms harboring gregarines when compared with mealworms which had been reared free from gregarines. This applied both to infected and non-infected larvae grown singly and in communal cultures. When, however, larvae were grown on a sub-optimal diet, the gregarines had a considerable effect on the final pupal weight and the ability of the larva to complete development.