bluuebunny/crossref_metadata_2025 huggingface dataset:
Demonstration and Preliminary Characterization of an Enzyme Capable of the Further Metabolism of the Thymidine Catabolite, β‐Aminoisobutyric Acid, in Tetrahymena pyriformis Strain GL1
Authors: NIEMANW MARILYN A. , BERECH JOHN
Published: May 2007

ABSTRACT. The enzyme that catalyzes the critical conversion of the proposed reductive thymidine catabolic end product, P‐aminoisobutyric acid, to the initial anabolic reutilization substrate, probably methylmalonic semialdehyde, was investigated to un‐ ambiguously substantiate the operation of a reductive pyrimidine catabolic and reutilization pathway in Tetrahymena pyriformis strain GL. Although most of the studies on the further metabolism of P‐aminoisobutyric acid in other organisms suggested transamination of p‐aminoisobutyric acid to methylmalonic semialdehyde followed by its further oxidation to methylmalonic acid, such a transaminating system could not be demonstrated. By means of a sensitive fluorometric assay system, however, a low, but significant amount of P‐aminoisobutyric acid oxidase activity was detected. This enzymatic activity exhibited the following characteristics in homogenates: good activity in alkaline 0.2 M Tris‐HCI buffer with a rather broad pH optimum ranging from 7.8 to 9.0; optimum activity at a temperature of 37°C; stimulation on the addition of exogenous FAD; inhibition on the addition of divalent cations, EDTA, or PCMB; little stimulation on the addition of detergents; and no increase in activity on repeated freezing and thawing. In addition, crude preparations of this oxidase were found to be relatively stable when stored up to one week either refrigerated or frozen, to have a specific activity of 2.8 nmoles/min/mg of protein, and to have a K, of 3.6 × 10− M for d,l‐β‐aminoisobutyric acid. Whether this high Km, is physiologically significant or not, however, will have to await further investigation. Preliminary (NH4)2SO4 fractionation and affinity chromatography studies also indicated that this enzyme appears to be a unique and specific oxidase whose activity is separable from other marker enzymes, including other oxidases.
Ionic Resistance in Strains of the Tetrahymena pyriformis Complex1
Authors: RICCI NICOLA
Published: May 2007

ABSTRACT. Strains of Tetrahymena thermophila were examined in an attempt to establish what role certain ions (Na+, K+, Li+, Ba++, Ca++, Mg++, Mn++, Al+++, Fe+++) play in influencing cell survival time in a culture medium. In short‐term experiments (20–30 min), cell survival time in a 1% peptone medium is directly related to the valence of the ion employed. Long‐term observations (lasting up to five days) in a 1% peptone medium containing lower ion concentrations revealed that the effects on cell‐cycle time are not correlated with the valence state of the ion. Comparisons were made among the ionic resistances of strains of T. thermophila, of T. pyriformis sensu stricto, and of two subspecies of T. pigmentosa. Strains within a species are highly correlated in their patterns of ionic response, while marked differences between species occur. The most distinctive group of strains examined came from one of the subspecies (syngen 6) of T. pigmentosa.
Occurrence of Amoebae In and Around the Mushroom Laccaria trullisata
Authors: NAPOLITANO JOSEPH J. , FLANAGAN VIRGINIA D.
Published: May 2007

ABSTRACT. Studies performed with the basidiomycete Laccaria trullisata collected from the sandy beach at the Hempstead Lake State Park, Long Island, New York, during the growing seasons of 1979 and 1980, have demonstrated a carposphere (equivalent to rhizosphere) effect. This region exerts a positive influence on the population density of amoebae when numbers are compared with those obtained in the bare sand 5 cm away. Moreover, amoebae have been shown to exist in, and have been recovered from, internal tissue of the cap (72%) and stalk (91%) of these mushrooms. A partial characterization of three strains of amoebae isolated from the internal tissue of L. trullisata and established in clonal culture is presented.
Oral Apparatus Structure in the Carnivorous Macrostomal Form of Tetrahymena vorax
Authors: SMITH HARRIETT E.
Published: May 2007

The structure of the oral apparatus in the carnivorous macrostomal form of Tetrahymena vorax has been investigated using serial thin sections and preparations of isolated oral apparatuses. The cilia of the oral apparatus are organized into an undulating membrane that borders the right and part of the posterior margin of the buccal cavity and three membranelles that project from plateaus on the anterior surface. Each membranelle consists of one short row and two longer rows of hexagonally packed kinetosomes. The organization of the microtubules of the oral ribs is identical to that in the T. vorax microstomal cell type. However, the first oral rib originates near the first kinetosome at the anterior end of the undulating membrane. The fine filamentous reticulum that underlies part of the oral ribs in the macrostomal cell type is not striated, unlike the reticulum in the microstomal form. A band of filaments similar to the fine filamentous reticulum extends around the anterior margin of the large cytostomal opening that occupies most of the posterior part of the oral cavity. The single row of microtubules along the left side of the oral cavity and cytostome also has filaments associated with it. A major difference between the microstomal and macrostomal forms in the structure of the oral apparatus is in the oral connectives. The macrostomal cell type contains only a single cross‐connective that joins the three membranelles and the anterior portion of the undulating membrane. The posterior or peripheral connective between the posterior ends of membranelles one and two and the posterior end of the undulating membrane is absent.
The Formation and Fate of Tetrahymena patula Cryptostomes
Authors: GABE PHILIP R. , WILLIAMS NORMAN E.
Published: May 2007

A reliable method for producing reproductive cysts in Tetrahymena patula is described. The procedure involves the isolation of macrostomes without cytopharyngeal pouches in microdrops of distilled water under oil. The study of silver‐impregnated specimens has shown that a complex pattern of oral resorption and reformation occurs within the cyst that leads to the formation of a group of small cells with recessed oral apparatuses. These cells, called “cryptostomes,” swim very rapidly on excystment and are incapable of either feeding or reproducing. They are presumably dispersal forms. Oral morphogenesis during the transformation of excysted cryptostomes into microstomes and macrostomes is also described.
Effects of Malaria (Plasmodium berghei) on the Maternal‐Fetal Relationship in Mice
Authors: ODUOLA A. M. J. , HOLBROOK T. W. , GALBRAITH R. M. , BANK H. , SPICER S. S.
Published: May 2007

Plasmodium berghei infection was more severe in pregnant than in nonpregnant mice. Infection initiated on gestation day 7 resulted in rapidly increasing parasitemia and deaths of all pregnant mice within 12 days, while some nonpregnant mice survived until day 21 postinfection. When mice were infected on gestation day 12 or 14, a proportion of mice died before parturition; but some animals survived to deliver living pups. Reduced birthweights and increased spleen weight to body weight ratios were seen in pups from infected mice as compared with pups from uninfected animals. Histopathological abnormalities of placentae from infected animals included degeneration of the normal labyrinthine architecture and thickening of the trophobast separating maternal and fetal blood vessels.
Phosphonoacetic Acid: Effect on and Disposition by Tetrahymena1
Authors: SMITH JOSEPH DONALD , KING JEFFERY C. , HOLLAND ERIC C.
Published: May 2007

The antiviral agent phosphonoacetic acid inhibits growth of Tetrahymena thermophila at concentrations comparable to those inhibiting growth of other eukaryotic cells, with 50% inhibition at 0.5 mM phosphonoacetic acid. The compound is cytotoxk to Tetrahymena at concentrations greater than 2.0 mM. When a culture of Tetrahymena the growth of which was totally inhibited by 2.0 mM phosphonoacetic acid was diluted with fresh medium, growth resumed in an exponential, rather than synchronous, fashion. [2–14C]phosphonoacetic acid is not metabolized by Tetrahymena.
Sensitivity and Specificity of the Indirect Fluorescent Antibody Test in the Study of Four Murine Coccidia1
Authors: TILAHUN G. , STOCKDALE P. H. G.
Published: May 2007

The sensitivity and specificity of the indirect fluorescent antibody (IFA) test for the detection of serum antibodies were examined in mice that were infected with Eimeria falciformis, E. ferrisi, E. papillata, or E. vermiformis. For the study of each species, five groups of mice were given graded inoculation doses of 10, 102, 103, 104, or 105 sporulated oocysts in a primary infection. The sixth group was infected with three sequential doses of 1.5 times 103, 1.5 times 104, and 1.5 times 105 sporulated oocysts per mouse at two‐ to three‐week intervals. All groups of infected mice developed serum antibodies. Sera were titrated by the IFA test with purified sporozoites. Strong fluorescence and high IFA titers were observed with homologous reactions mainly with the sera from mice infected with the higher inoculation dose levels in primary infections and from those given three sequential inoculation doses. Immunological cross reaction among the four species of Eimeria occurred at dilutions of 1:10 to 1:160. Very weak or no fluorescence of free sporozoites was observed with sera from noninfected mice, and there was no fluorescence of sporozoites contained in intact sporocysts.
A Light and Electron Microscopical Study of Trichoduboscqia epeori Léger (Microspora: Duboscqiidae)1
Authors: BATSON B. S.
Published: May 2007

Trichoduboscqia epeori Léger was found to parasitize nymphs of the mayfly Rhithrogena iridina Kolenati in southwest Germany for a new host record. It was studied by light and electron microscopy. The pansporoblast membrane is evaginated at several points, usually four, to produce long needle‐like appendages >20 μm in length with a resilient inner core superficially resembling collagen, which is thought to maintain their orientation. It is suggested that the pansporoblast appendages may play a role in host infection. The structure and ultrastructure of developmental stages are recorded for the first time. Apart from the pansporoblast appendages, the ultrastructure of T. epeori conforms to the general pattern seen in many other pansporoblastic Microspora. Typically 16 spores are produced per pansporoblast but 32‐spore pansporoblasts were also found, and the taxonomic significance of this is discussed.
Obtention de Polyplastron multivesiculatum (Cilié Entodiniomorphe du Rumen) en Condition Axénique1
Authors: BONHOMME ANNIE , FONTY GERARD , SENAUD JEAN
Published: May 2007

RESUME

Une méthode d'obtention et de maintien des Ciliés du rumen (genre Polyplastron) en culture axénique, est décrite. Elle comporte deux périodes d'incubation en anaérobiose avec différentes associations d'antibiotiques, séparées par un lavage et un renouvellement du milieu nutritif. Dans ces conditions. Polyplastron multivesiculatum est obtenu à l'état axénique et maintenu en survie pendant cinq jours.

A method to obtain and keep rumen ciliates (genus Polyplastron) in an axenic condition is described. It consists of two incubating periods with different antibiotic mixtures, separated by washing and renewing of nutrient medium. Under these conditions, Polyplastron multivesiculatum is established in an axenic state and can survive thusly for five days.
Electrophoretic Analysis of Endonuclease‐Generated Fragments of k‐DNA, of Esterase Isoenzymes, and of Surface Proteins as Aids for Species Identification of Insect Trypanosomatids1
Authors: CAMARGO E. PLESSMANN , MATTEI D. M. , BARBIERI CLARA L. , MOREL C. M.
Published: May 2007

In order to verify the applicability of biochemical methods for species identification of Trypanosomatidae, 13 species of monoxenic trypanosomatids plus the heteroxenous Trypanosoma cruzi were comparatively analyzed by three different biochemical methods. Insect trypanosomatids examined were: Crithidia acanthocephali, C. fasciculata (three varieties), C. luciliae luciliae, C. luciliae thermophila, C. deanei, C. oncopelti, Herpetomonas muscarum muscarum, H. megaseliae, H. samuelpessoai, H. mariadeanei, Leptomonas seymouri, L. collosoma, L. samueli, and Blastocrithidia culicis. Also included in the survey were aposymbiotic strains of C. deanei and C. oncopelti. Methods used were: electrophoretic profiling of endonuclease‐generated fragments of k‐DNA, esterase isoenzymes profiling, and polyacrylamide‐gel electrophoresis (SDS‐PAGE) of radioiodinated cell surface proteins. Interspecific but not intraspecific differences were detected by all three methods among the 13 monoxenic species examined. Thus, it is concluded that these methods can be successfully used, in addition to classical criteria, for species identification of insect trypanosomatids.
The Behavior and Fine Structure of the Dorsal Bristles of Euplotes minuta, E. aediculatus, and Stylonychia mytilus (Ciliata, Hypotrichida)1
Authors: GÖRTZ HANS‐DIETER
Published: May 2007

ABSTRACT. Studies of the bristle (dorsal) cilia of Euplotes minuta. E. aediculatus, and Stylonychia mytilus by light and electron microscopy indicate that these cilia do not beat metachronously in any of the species. The bristle cilia in Stylonychia may beat actively, but those in Euplotes stand erect or are bent in different directions with the flow of water. The duration and degree of bending appear correlated with the duration and velocity of the water current. The fine structure of the bristle complex is similar in both Euplotes species and like other reports of Euplotes in the literature. The complex consists of paired kinetosomes, the anterior bearing a short cilium containing four to six rows of fibrous balls (lasiosomes) oriented along the anterior surface of the axoneme, the posterior lacking a cilium but with a small cap. Microtubular ribbons are associated with the paired kinetosomes, and a collar with a pronounced alveolar ring underneath the pellicular membrane tightly surrounds the cilium at the opening of the bristle pit. The bristle complex in S. mytilus differs from that of Euplotes and other hypotrichs in that it has a single kinetosome in interphase cells and, attached to the kinetosome, a prominent fibrous structure (parakinetosomal body). Microtubules are attached to the parakinetosomal body. As in Euplotes, the bristle unit is surrounded by mucocyst‐like organelles (ampules). Observations of behavior and fine structure suggest that the dorsal bristles may be sensory, perhaps responding to stimuli from water currents, although other functions are possible, too.
Physiological Studies on the Cytopharyngeal Pouch, a Prey Receptacle in the Carnivorous Macrostomal Form of Tetrahymena vorax1
Authors: SHERMAN GARY B. , BUHSE HOWARD E. , SMITH HARRIETT E.
Published: May 2007

ABSTRACT. Various ions and treatments known to alter the availability of free Ca2+ were examined with respect to their effects on the cytopharyngeal pouch, a large prey receptacle found in the potentially carnivorous macrostomal form of Tetrahymena vorax. Addition of Ca2+, Ba2+, or Sr2+ induced the pouch to separate from the region of the cytostome, forming a large empty vacuole. Na+, alone, had no effect, but lowered the concentration of Ca2+ required to produce maximum vacuolar formation in populations of cells. Vacuolar induction was also initiated by the cation ionophore A23187 or by subjecting macrostomal cells to an electric current. In the presence of divalent cation chelators EDTA and EGTA, the cytopharyngeal pouch collapsed and was resorbed. Taken together, these results suggest that Ca2+ plays an important role during phagocytosis in this cell type.
ATPase‐Associated Oligomycin Resistance in Acanthamoeba castellanii1
Authors: SEILHAMER JEFFREY J. , BYERS THOMAS J.
Published: May 2007

ABSTRACT. The multiplication rate of “wild‐type” (WT) populations of Acanthamoeba castellanii was inhibited 50% by ∼3 μg otigomycin/ml; OliR2 an oligomycin resistant cell line, required ∼27 μg/ml for the same inhibition. ATPase solubilized from OliR2 mitochondrial fractions required 3–10‐fold higher concentrations of oligomycin than did identical WT fractions to achieve 50% inhibition of activity. Resistance was correlated with altered mitochondrial ATPase sensitivity to oligomycin.
Comparison of Six Isoenzymes from 10 Species of Crithidia1
Authors: DE LIMA VILNEYDE M. Q. GONCALVES , ROITMAN ISAAC , ALVES CYRENE DOS SANTOS
Published: May 2007

ABSTRACT. The electrophoretic mobility of six isoenzymes from 10 species of Crithidia from insects were compared. Five zymogram patterns emerged. Pattern I was presented by C. acanthocephali and Crithidia sp. from Euryophthalmus davisi; pattern II by C. hutneri and C. luciliae thermophila; pattern III by C. arili; pattern IV by C. luciliae luciliae, Crithidia sp. from Aedes solicitans, C. harmosa, and C. fasciculata; pattern V by C. oncopelti.
Surface Changes Induced by Immune Serum on Eimeria tenella Sporozoites and Merozoites
Authors: WITLOCK DALE R. , DANFORTH HARRY D.
Published: May 2007

ABSTRACT. The surface of merozoites and sporozoites of Eimeria tenella was affected by incubation with E. tenella‐immune chicken serum (ICS). Normal chicken serum (NCS) and heat‐inactivated ICS had no effect on the pellicular surface of either developmental stage. Sporozoites formed surface bulges or swellings after 10 min of incubation with ICS, and by 15 min postincubation, the morphology of the sporozoites was distorted by a surface coating of fibrinous material. Merozoites exposed to ICS were similarly coated, but surface swelling was not as severe. The coating formed rapidly and was seen as early as 5 min postincubation. Sporozoites incubated with heat‐inactivated ICS supplemented with normal chicken serum were coated with a fibrinous material and in some cases lysed. These data indicated that complement must be present for the surface interaction to occur.
The Fine Structure of Acanthamoeba astronyxis, with Special Emphasis on Encystment1
Authors: LASMAN M.
Published: May 2007

ABSTRACT. The fine structure of the trophozoite, encysting cells, and the cyst of Acanthamoeba astronyxis has been examined. In the trophic form a microtubule organizing center was associated with a well developed Golgi complex. During encystment the organelles of the amoeba changed considerably. The profiles of rough endoplasmic reticulum elongated and were often arranged in circles of multilayered concentric systems, enclosing mitochondria, the nucleus, or other inclusions. The mitochondria showed a tendency toward elongation and constriction. One or two nucleolus‐like bodies appeared in the nucleus. Lipid droplets increased considerably in amount and were distributed individually or as aggregates. The mature cyst was star‐shaped and surrounded by an almost circular exocyst and an endocyst that was closely apposed to the cell membrane. Both walls differed in their thickness and granulation. The exocyst was continuous over the entire cyst, while the endocyst was interrupted by gaps, ostioles. in the region of the rays. Within the ostioles was a bell‐shaped structure, the operculum. The latter was composed of a granular material comparable in electron density to that of the endocyst.
The Cysts of Blastocrithidia triatomae Cerisola et al., 19711
Authors: PENG PHILLY LIN‐MOO , WALLACE F. G.
Published: May 2007

ABSTRACT. Flagellar cysts of Blastocrithidia triatomae form from active flagellates by diminution in size. The pellicular microtubules disappear. The inner layer of the cell membrane thickens progressively as the organism shrinks. The fully formed cyst has an electrondense layer that corresponds to the outer layer of the unit membrane. An electron‐lucent layer is approximately twice the thickness of the middle layer of the unit membrane. Inside that is a 92 nm layer that may represent the cytoplasm. The nuclear content is in the form of whorled bundles of 10–15 nm fibrils. The kinetoplast was not seen in electron micrographs of cysts.
The Lipid Requirement of the Ciliated Protozoon Tetrahymena patula
Authors: LUKSAS ANTHONY J. , ERWIN J. A.
Published: May 2007

ABSTRACT. The ciliated protozoon Tetrahymena patula can be grown in a chemically defined medium supplemented with a suitable lipid. High purity natural phosphoiipids; mono‐, di‐, and triglycerides: and free fatty acids are suitable lipids. The more complete lipids appear to serve simply as nutritionally convenient sources of fatty acids. T. patula can also be grown in the synthetic medium supplemented with cholesterol or other sterols in lieu of fatty acid containing lipids. Supplementation with either ethanolamine or choline permits suboptimal growth of the ciliate in a lipid‐free synthetic medium. No other water soluble compound, of a variety that were tested, permitted growth of the ciliate in the lipid‐free medium.
An Inhibition and Kinetic Study of Acid Phosphatase in Paramecium caudatum and Paramecium tetraurelia1
Authors: FOK AGNES K.
Published: May 2007

ABSTRACT. Inhibition, inactivation, pH, and kinetic studies using both homogenates and purified lysosomal fractions of Paramecium caudalum and of P. tetraurelia were carried out to examine the lysosomal acid phosphatase (AcPase) and its relationship to p‐nitrophenylphosphatase (pNPPase), glucose‐6‐phosphatase (G6Pase), and 5′‐nucleotidase (AMPase). The results generally support the idea that Paramecium cells contain a distinct lysosomal AcPase with a broad substrate specificity. The hydrolysis of glucose‐6‐phosphate (G6P) and adenosine 5′‐monophosphate (AMP) was shown to be due to this enzyme, suggesting that true G6Pase and AMPase may be lacking in these two species; however, some hydrolysis of AMP at pH 7.5 catalyzed by an unknown soluble enzyme distinct from alkaline phosphatase and Na+‐K+‐ATPase was observed. Since the hydrolysis of p‐nitrophenylphosphate (pNPP) at acid pH was also shown to be due to AcPase alone, pNPPase could be used as a rapid assay for Paramecium AcPase. At an alkaline pH, however, this activity was catalyzed by an alkaline phosphatase located in the cytosol fraction. P. caudatum AcPase was shown to have kinetic properties similar to those of purified rat liver and human prostatic AcPase and to have relative substrate affinities in the order of G6P < β‐glycerophosphate < pNPP < AMP. These different substrate affinities might account for the observed differences in the inhibition of the four lysosomal activities by NaF, L(+)‐tartrate, and molybdate, all of which inhibited the hydrolysis of G6P, β‐glycerophosphate, and pNPP competitively, but which exhibited a noncompetitive inhibition of a mixed type with the hydrolysis of AMP.
Cyclic Adenosine 3′,5′‐Monophosphate Binding Protein in Tetrahymena: Properties and Subcellular Distribution1
Authors: KUDO SHUZO , NOZAWA YOSHINORI
Published: May 2007

ABSTRACT. Cyclic AMP binding activity was determined in the ciliate Tetrahymena pyriformis NT‐1 strain. The fractions having the binding activity were eluted in a single peak coincident with a protein kinase activity. Although metal ions were not essential for activity, the binding was slightly activated by Mg2+ or Ca2+. The binding activity was sensitive to temperature, ionic strength, and pH of the reaction mixture and was decreased by treatment of the cytosol protein with trypsin or by heating at 100°C. The binding was specific for cyclic AMP, with an estimated apparent Kd of 40 nM. When the cyclic AMP binding activity in subcellular fractions was measured, an increase in the activity of ciliary, mitochondrial, and microsomal fractions was observed during the transition from the exponential to the stationary phase of cell growth, whereas no significant change occurred in the binding activity of the whole cell homogenate. These results suggest that the redistribution of cyclic AMP binding proteins may be implicated in the regulation of cyclic AMP concentration in the cell.
Enzymes of de novo Pyrimidine Biosynthesis in Babesia rodhaini1
Authors: HOLLAND JOHN W. , GERO ANNETTE M. , O'SULLIVAN WILLIAM J.
Published: May 2007

ABSTRACT. The pathway of de novo pyrimidine biosynthesis in the rodent parasitic protozoa Babesia rodhaini has been investigated. Specific activities of five of the six enzymes of the pathway were determined: aspartate transcarbamylase (ATCase: E.C. 2.1.3.2): dihydroorotase (DHOase: E.C. 3.5.2.3): dihydroorotate dehydrogenase (DHO‐DHase: E.C. 1.3.3.1); orotate phosphoribosyltransferase (OPRTase: E.C. 2.4.2.10); and orotidine‐5′‐phosphate decarboxylase (ODCase: E.C. 4.1.1.23). Michaelis constants for ATCase, DHO‐DHasc. OPRTase, and ODCase were determined in whole homogenates. Several substrate analogs were also investigated as inhibitors and inhibitor constants determined. N‐(phosphonacetyl)‐L‐aspartate was shown to be an inhibitor of the ATCase with an apparent K, of 7μM. Dihydro‐5‐azaorotate inhibited the DHO‐DHase (K, 16 μM) and 5‐azaorotate (Ki, 21 μM) was an inhibitor of the OPRTase. The UMP analog, 6‐aza‐UMP (Ki, 0.3 μM) was a potent inhibitor of ODCase, while lower levels of inhibition were found with the product. UMP (Ki, 120 μM) and the purine nucleotide, XMP (K1, 95 μM). Additionally, menoctone, a ubiquinone analog, was shown to inhibit DHO‐DHase.
A Note on the Possible Ecological Significance of Chemotaxis in Certain Ciliated Protozoa1
Authors: ANTIPA GREGORY A. , MARTIN KATHLEEN , RINTZ MICHAEL T.
Published: May 2007

ABSTRACT. A heat‐stable chemoattractant has been isolated from bacterial cultures. This component has a molecular weight in the range of 500–1000 daltons, is produced by both Gram‐positive and Gram‐negative bacteria, and serves equally well as an attractant for both the bacterial feeding Paramecium and for its natural predator, Didinium. Aspects of the ecological relationship between bacterial feeding ciliates and their ciliate predators are briefly discussed with respect to responses of both predator and prey to such a common chemotactic bacterial factor.
Herpetomonas megaseliae, Crithidia fasciculata, and Leptomonas collosoma (Kinetoplastida, Trypanosomatidae) in Feces of Lizards Fed Culture Forms of the Flagellates
Authors: DOLLAHON NORMAN R. , HAGER RICHARD , HUA TA CHIH
Published: May 2007

ABSTRACT. Herpetomonas megaseliae, Crithidia fasciculata, and Leptomonas collosoma from culture survived gut passage in Anolis carolinensis following their ingestion by this lizard. Maximum persistence of H. megaseliae in lizards, as detected by fecal culture, was seven days. No invasion of tissues by H. megaseliae could be detected by means of sectioned material, stained impression slides, or cultures inoculated with material from organs. Crithidia fasciculata was evident in cloacal fluid for up to three days in wet mount preparations. Leptomonas collosoma was observed in feces 24 h after the organisms were fed to lizards. Both C. fasciculata and L. collosoma were cultured from feces of lizards fed the parasites 24 h earlier. Herpetomonas megaseliae was differentiated in lizard feces, with greater than 40% of the forms observed being paramastigotes or opisthomastigotes. Truncate, semispherical forms resembling choanomastigotes were seen, but the kinetoplast was posterior to the nucleus in some of these. Many forms showed extensive coiling of the axoneme within the body of the flagellate. Choanomastigotes and spheromastigotes of C. fasciculata and promastigotes, sphero‐mastigotes and amastigotes of L. collosoma were also observed in the feces.
Methods for the Large‐Scale Cultivation of an Oxytricha (Ciliophora: Hypotrichida)1
Authors: LAUGHLIN TOMMIE J. , HENRY JOHN MICHAEL , PHARES EDWIN F , LONG MARY V. , OLINS DONALD E.
Published: May 2007

ABSTRACT. Oxytricha strains used in biochemical studies have traditionally been grown in unaerated, unagitated culture tubes or Fernbach flasks. These cultures are limited in volume to about one liter and have a very nonuniform distribution of cells, with the majority of the cells at the very top or bottom of the medium. We have found conditions in which Oxytricha can be grown in 50‐liter fermentation vats. The cultures grow to a uniform density of about 6000 cells/ml.
Morphologie et Ultrastructure de Spirochona gemmipara Stein, 1852 (Ciliophora, Chonotrichida). II. Appareil Cytoproctal et Système Excréteur de I'Adulte1
Authors: FAHRNI JOSE F.
Published: May 2007

RESUME. Dans la région antérieure de Spirochona gemmipara, l'emploi combiné de la microscopie à contraste interférentiel (No‐marski), des colorations au protargol et de la microscopie électronique à transmission révèle l'existence d'un appareil cytoproctal et d'un système excréteur (“contractile vacuole complex”), lesquels ont souvent été confondus entre eux ou avec le cytopharynx. L'appareil cytoproctal comprend un orifice externe situéà la base de la collerette, un canal cytoproctal long d'environ 20 μm limité par une pellicule alvéolaire, et le cytoprocte proprement dit. Le système excréteur est formé de 6 à 8 canaux sinueux pouvant mesurer jusquà 20 μm de long et s'ouvrant chacun par un pore situé sur la face interne de la collerette, parmi la ciliature. L'ostium, c'est‐à‐dire l'orifice profond de chaque canal, est en relation avec une vacuole dans laquelle aboutit un spongiome tubulaire bien développé. Bien que profondément enfoncés dans le cytoplasme, le cytoprocte et les ostia du spirochone ne semblent pas fondamentalement diffêrents des organites correspondants décrits chez Paramecium et chez Tetrahymena.
Ultrastructural Studies of a Vesicle System Associated with Endoplasmic Reticulum in Exo‐Erythrocytic Forms of Plasmodium berghei1
Authors: MEIS J. F. G. M. , JAP P. H. K. , VERHAVE J. P. , MEUWISSEN J. H. E. T.
Published: May 2007

ABSTRACT. Fine structural studies of a specialized vesicle system associated with the endoplasmic reticulum (ER) of exo‐erythrocytic Plasmodium berghei suggest that this system may be the equivalent of a Golgi apparatus. Patches of ER, randomly distributed in the cytoplasm of developing parasites, are formed of smooth and ribosome‐studded cisternae intermingled with each other. The vesicle systems are located between as well as at the edges of ER aggregates and appear to be in different stages of budding from the cisternae. Prolonged osmication reveals distinct staining of the nuclear envelope and ER of the parasites as well as part of the Golgi apparatus of the hepatocytes. However, the small vesicles associated with the parasite's ER are unstained, as are the coated vesicles in the Golgi region of the liver cell. These sites in the parasite cytoplasm seem comparable to the concave surface of the Golgi apparatus in liver cells. The pinched‐off vesicles fuse with others to form the prominent peripheral vacuolization characteristic of the nearly mature exo‐erythrocytic form. The formation of these peripheral vacuoles and their subsequent fusion with the parasite membrane may be an exocytosis mechanism supplying the rapidly expanding parasite with new plasma membrane material.
Paramecium sonneborni n. sp., a New Member of the Paramecium aurelia Species‐Complex1
Authors: AUFDERHEIDE KARL J. , DAGGETT PIERRE‐MARC , NERAD THOMAS A.
Published: May 2007

ABSTRACT. From an intermittent stream in College Station, Texas, a Paramecium was isolated that did not appear to belong to any recognized species. On the basis of nuclear and whole‐body morphology, it can be assigned to the Paramecium aurelia species‐complex, and it can be distinguished from other members of that complex on the basis of mating‐type reactivity and isoenzyme patterns. These characteristics are felt sufficient to justify a new species assignment. The new species has been named Paramecium sonneborni n. sp. in honor of the late Dr. Tracy M. Sonneborn of Indiana University.
Comparison of the Esterases and Acid Phosphatases in Paramecium multimicronucleatum, Syngens 1–5, P. jenningsi, P. caudatum, and the P. aurelia Complex1
Authors: ALLEN SALLY LYMAN , NERAD THOMAS A. , RUSHFORD CAROLINE L.
Published: May 2007

ABSTRACT. Forty‐eight stocks in Paramecium jenningsi, syngens 1–5 of P. multimicronucleatum, P. caudatum, P. primaurelia, P. biaurelia, and P. tetraurelia were grown axenically and tested for their esterases and acid phosphatases using starch gel electrophoresis. The five esterases and the acid phosphatases previously characterized in species of the P. aurelia complex were also found in P. jenningsi, and three to four of the esterases and the acid phosphatases were found in the P. multimicronucleatum species complex and in P. caudatum. Additional subtypes were observed for each of the enzyme phenotypes in these new (though here unnamed) species of Paramecium. Two of the new acid phosphatase subtypes, which depart radically in mobility and in pattern, were found in syngen 3 of P. multimicronucleatum and in P. caudatum. Except for syngens 1 and 5 in P. multimicronucleatum, the degree of similarity between syngens 1, 5 and 2, 3, and 4 appears to be very low—perhaps even lower than that seen for species in the aurelia complex. More realistically, the syngens of P. multimicronucleatum should be considered as separate species although they are not here given separate taxonomic names. Limited sharing of subtypes occurred between species in different species complexes. This observation suggests that the molecular distances between species complexes may be even greater than between species within a complex.
Scanning Electron Microscopy of the Adoral Ciliary Zone of Entodinium Stein (Ciliophora, Entodiniomorphida)1
Authors: IMAI SOICHI , TASHIRO HIDETOSHI , ISHII TOSHIO
Published: May 2007

The adoral ciliary zone of the rumen ciliates, Entodinium spp., was observed topographically in the SEM. The upper side of the anterior end of the body was indented by the vestibulum, which had cilia arranged on its right wall and ribs along the left wall. The adoral ciliary zone could be divided into at least two arrangements. The outer ciliary zone had many membranelle‐like structures, which consisted of cilia arranged radially from the body axis. Each membranelle‐like structure consisted of two rows of about eight cilia each lining up in a single file. It was, however, different from a typical membranelle, because its cilia were connected with the vestibular cilia and were arranged not spirally but on a plane. These cilia extended toward the outside because of the projecting cytoplasm from which they originated. In contrast, the cilia of the inner ciliary zone were aggregated to form relatively unsystematic bundles. Since the vestibular opening was slanted on the upper side of the body, the ciliary bundles were thick on the lower side and sparse on the upper side of the body. Neither outer nor inner ciliary zones completely surrounded the vestibular opening. The ciliature started from the left side of the vestibular opening, encircled the lower side of the body, and entered the vestibulum from its right side. The functions of these two types of ciliary arrangements are discussed.
Ultrastructural Aspects of the Somatic Cortex and Contractile Vacuole of the Ciliate, Ichthyophthirius multifiliis Fouquet1
Authors: CHAPMAN GEORGE B. , KERN ROBERT C.
Published: May 2007

The trophont stage in the life cycle of Ichthyophthirius multifiliis was studied in the electron microscope. Surface ridges contain up to 24 ridge microtubules, disposed as a ribbon. Kinetosomes show the classic morphology of 9 triplets of microtubules. Associated with each kinetosome is a kinetodesmal fibril, originating in proximity to triplets 5, 6, and 7, and having a 30 nm periodicity; 3 to 5 postciliary microtubules, originating between triplets 8 and 9; and up to 3 transverse microtubules, originating at triplet 4, as well as a parasomal sac. Each cell is partially enclosed by a system of 3 “unit” membranes: the outer limiting membrane, and the outer and inner alveolar membranes. The last two membranes define the alveolar sac. Mucocysts, each with a dense core, are present in large numbers. The contractile vacuole system includes the contractile vacuole, associated tubules and vesicles, injection canals, a discharge canal, and a pore. Microtubules abound in the walls of the contractile vacuole, injection and discharge canals, and in the region of the pores, where both ring and radial microtubular arrangements are noted. The ultrastructure suggests that I. multifiliis is more closely related to Tetrahymena pyriformis than to Paramecium aurelia.
Nucleosome Phasing in Tetrahymena Macronuclei1
Authors: PRATT KAREN , HATTMAN STANLEY
Published: May 2007

Core‐protected DNA can drive only 60% of the Tetrahymena thermophila macronuclear genome into duplexes in hybridization experiments. This core‐protected DNA therefore contains only a subset of the genome complexity. We interpret this to mean that a large fraction, if not all, of the genome is phased with respect to nucleosome placement. Among the sequences present in total DNA and absent from core‐protected DNA are most of the sequences containing N6‐methyladenine (MeAde) residues, consistent with our previous demonstration that most of these residues lie in linker DNA. We show that these results are not due to artifacts resulting from the small size of the DNA driver, nor are they due to any sequence preferences exhibited by staphylococcal (staph) nuclease. This is the first evidence that nucleosome phasing may be a bulk genome characteristic.
Cortical Control over the Direction of Macronuclear Elongation in the Heterotrich Ciliate Stentor coeruleus1
Authors: TERRA NOËL DE
Published: May 2007

Earlier experimental work involving macronuclear implants in Stentor coeruleus has shown that the cytoplasmic cortex of the nuclear site 1) attracts the macronucleus and 2) holds it in place during interphase. Now experiments indicate macronuclei transferred with overlying cortex elongate in the direction of the transferred cortical pigment stripes, whether or not the transferred stripes realign in the direction of the host stentor's stripes. Therefore the third function of the cortex is to determine the direction of elongation and thus assure that both daughter cells at division receive part of the macronucleus.
Ultrastructural Study on the Development of Babesia equi (Coccidia: Piroplasmia) in the Salivary Glands of its Vector Ticks1
Authors: MOLTMANN ULRICH G. , MEHLHORN HEINZ , SCHEIN EBERHARD , VOIGT WOLF P. , FRIEDHOFF KARL T.
Published: May 2007

The formation of Babesia equi sporozoites in the salivary glands of three tick species (Hyalomma anatolicum anatolicum, H. a. excavatum, Rhipicephalus turanicus) was studied by electron microscopy. The development was identical in all three vectors. On the 8th day post repletionem kinetes of B. equi had invaded alveoli of the nymphal salivary glands and were transformed to sporonts bounded by a single membrane. The sporonts were polymorphous bodies each with a highly lobed nucleus and numerous mitochondria. These stages persisted during ecdysis of the tick nymph to the adult stage. After attachment of these newly molted adults to a new host the formation of sporozoites was completed within five days. The sporonts occupied most of the infected alveolus and were extensively divided into cytoplasmic portions of various size. On the 4th day after attachment of the tick, sporozoite‐anlagen, into each of which a nucleus and a mitochondrion were incorporated, appeared at the periphery of the sporonts. An apical complex with a polar ring, rhoptries, and micronemes was formed at the tip of each protruding anlage. Finally thousands of pyriform sporozoites (3.0 × 1.2 μm) filled the hypertrophied alveolus. This development is similar to sporogony in the genus Theileria.
The Occurrence of Argentophilic Bands (“Virgulenes”) on the Cortical Surface of Tetrahymena1
Authors: READEY MARIJO A. , MCARDLE EUGENE W. , KOO JOO O.
Published: May 2007

When Tetrahymena furgasoni (W) are stained by the silver impregnation technic. a relatively small percentage of them show cortical argentophilic bands or stripes that are intermeridional in position. The number and arrangement of the bands as well as their distinctive appearance and abrupt daily peaks of occurrence render them distinct from other recognized components of the argyrome. The term “virgulene” is proposed as a name for this feature.
Cross‐reactivity of Vector‐borne Metacyclic Forms of Trypanosoma cruzi with Mammalian and Culture Stages1
Authors: VILLALTA FERNANDO , KIERSZENBAUM FELIPE
Published: May 2007

Metacyclic forms of Trypanosoma cruzi isolated from the hindgut of infected insect vectors (Rhodnius prolixus) were found to be immunologically cross‐reactive with cultured epimastigote, amastigote, and metacyclic stages of the parasite as well as with bloodstream trypomastigote forms by direct agglutination and indirect immunofluorescence techniques. Sera specific for each of these forms of the parasite systematically yielded maximal antibody titers when measured against the homologous antigen, indicating that antigenic determinants are shared by all of the developmental forms used in this work. Supporting this conclusion were the significant reductions in anti‐insect‐derived metacyclic antibody titer caused by absorption with any of the other life stages of T. cruzi. These results are relevant to the potential use of laboratory‐grown forms of T. cruzi in vaccination against a natural infection with this parasite.
Antigenic Differentiation of the Kinetoplastids Leishmania braziliensis and Trypanosoma cruzi by Means of Monoclonal Antibodies1
Authors: CONSTANTINE NIEL T. , ANTHONY RONALD L.
Published: May 2007

BALB/c mice were hyperimmunized with non‐infectious extracts of either Leishmania braziliensis promastigotes or Trypanosoma cruzi epimastigotes. When spleen cells from these mice were fused with P3X63Ag8 plasmacytoma cells, the resultant hybridomas synthesized monoclonal antibodies which displayed specific reactivity by indirect immunofluorescence with distinct subcellular components of the parasites. These studies revealed that antigens associated with the flagellum and with a nongranular component of the cytoplasm would account for much of the serologic cross‐reactivity observed between the two species. Conversely, antigens associated with surface and/or cytoplasmic granules and with an intracellular organelle believed to be the kinetoplast appeared to be species‐specific.
Comparison of Naegleria fowleri and Naegleria gruberi Cultivated in the Same Nutrient Medium1
Authors: CLINE M. , MARCIANO‐CABRAL F. , BRADLEY S. G.
Published: May 2007

The human pathogenic amoeboflagellate Naegleria fowleri and the nonpathogenic species N. gruberi can be cultivated axenically but usually in different media. Naegleria fowleri 6088 has been adapted to grow in Balamuth H‐4 medium, usually used to propagate N. gruberi nB81. and nB81 has been adapted to grow in supplemented Nelson's medium, usually used to propagate N. fowleri. N. gruberi nB81. grown in either medium, enflagellated 135 to 150 min after subculture to non‐nutrient amoeba saline, whereas 6088 required 225 min. Naegleria gruberi nB81 grown in either medium was agglutinated by 100 ug concanavalin A/ml, whereas N. fowleri 6088 was not. Naegleria fowleri and N. gruberi grown in Nelson's medium became rounded to a greater extent upon chilling at 5° C and remained rounded longer than Naegleria grown in Balamuth medium. The specificity of the surface antigens was an inherent characteristic of each species and not dependent upon the propagating medium. but Naegleria grown in Nelson's medium was agglutinated more reproducibly and more effectively by antiserum. N. gruberi was somewhat more resistant to acriflavine, actinomycin D, cycloheximide, or tetracycline than N. fowleri, regardless of the culture medium. Naegleria fowleri 6088 grown in Nelson's medium, however, was more resistant to actinomycin D, daunomycin. mithramycin. sulfamethoxazole, or tyrocidine than 6088 grown in Balamuth medium. There are limitations on the validity of comparisons of N. fowleri and N. gruberi based upon cultures grown in different media.
Ultrastructure of the Amoeboflagellate Tetramitus rostratus1
Authors: BALAMUTH WILLIAM , BRADBURY PHYLLIS CLARKE , SCHUSTER FREDERICK L.
Published: May 2007

The life‐cycle of the amoeboflagellate Tetramitus rostratus includes amoeboid, cyst, and flagellate stages. The ultrastructure of these three stages is illustrated, with particular emphasis on flagellate morphology. Amoeba morphology is typical of that of limax amoebas. Cysts, forming from trophic amoebas, are enclosed by a wall made up of two layers: ectocyst (ca. 70 nm), and endocyst (200 nm). The wall apparently forms from precursor material present in vesicles in the pre‐cyst stage cytoplasm. Flagellate morphology is characterized by a well‐defined top‐shaped profile, maintained by microtubules under the plasma membrane. The flagellar apparatus or mastigont consists of four flagella, their basal bodies, sheaves of microtubules associated with two of the basal bodies, and several rhizoplasts (periodicity 20 nm). A deep, microtubule‐supported, ventral invagination appears to function as a gullet. A small number of mitotic stages observed in amoeboid and flagellate individuals suggests similarity in the division process in both stages: intranuclear mitotic apparatus, nucleolus persisting through mitosis, no centrioles or basal bodies functioning as centrioles, difficulty in resolving chromosomes. The text compares ultrastructures of several amoeboflagellate organisms and evaluates the phylogenetic significance of those features common to different species. On the basis of this study, Tetramitus most closely resembles Naegleria spp.
Tandemly Repeated Sequence Families in Micronuclear DNA of the Ciliate Stylonychia pustulata1
Authors: DAWSON DEAN , STETLER DEBORAH J. , SWANTON MARSHAL T. , HERRICK GLENN
Published: May 2007

One third of a collection of cloned Stylonychia pustulata micronuclear DNA PstI fragments were found to be of a similar size, consistent with their being members of a repetitious sequence family with a repeat size of about 160 base pairs. Cross‐hybridization experiments confirmed that these small cloned fragments are related by sequence homology. Hybridization of the cloned repetitious sequences to PstI digested micronuclear DNA revealed a “ladder” of bands (step size = 160 base pairs), indicating that the repeats are found in tandem arrays. This is the first demonstration of highly repetitious, tandemly repeated sequences in a ciliated protozoan.