ALBERT

Notes: A method of delineating the extent of floods has been devised involving the establishment of flood surfaces and the creation of photogrammetric digital terrain models. These two data sources are combined in a computer aided map production system and comparisons are drawn between the efficacy of different forms of terrain modelling in a floodplain environment.〈section xml:id="abs1-2"〉〈title type="main"〉RésuméOn a mis au point, en vue de déterminer l'étendue d'innondations, une méthode numérique combinant les notions de surface innondable et de modèle numérique de terrain (MNT); on se pose le problème du choix du MNT dans un environnement de plaine innondable.〈section xml:id="abs1-3"〉〈title type="main"〉ZusammenfassungAngabe eines Verfahrens zur Ableitung der Ausdehnung von Überschwemmungen, wobei die Flut-Oberfläche bestimmt und photogrammetrisch digitale Geländemodelle erzeugt werden. Diese beiden Datenquellen werden in einem rechnergestützten Kartiersystem kombiniert, und es werden Vergleiche bezüglich der Wirksamkeit verschiedener Verfahren der Geländemodellierung in Flussniederungen gezogen.

Notes: SYNOPSIS. A chemically defined culture medium has been developed for the soil amoeba Hartmannella rhysodes Singh which contains the minimum essential organic requirements for growth. The medium consists of 7 amino acids, 3 vitamins, a carbon source (e.g. glucose) and inorganic salts.

Notes: SYNOPSIS. A mutant strain of Astasia longa utilized glucose for growth whereas the parent (J) strain did not. The optimal pH for growth of the mutant with glucose (sole carbon source) was near neutrality; the optimal glucose concentration 0.02 M. Cell-free extracts or cell homogenates produced C14O2 when incubated in the presence of C14-labeled glucose. On the other hand, after incubation with C14-labeled glucose, intact parent cells and their respiratory CO2 showed no radioactivity while the mutant-strain cells and CO2 produced were active. Dissimilation of glucose-1-C14 and glucose-6-C14 yielded the same amount of radioactivity in metabolic CO2 in cell-free extracts of both strains. Of five enzymes assayed, hexokinase, phosphoglucomutase, and lactic dehydrogenase were present whereas glucose-6-PO4 dehydrogenase and glucose dehydrogenase were absent in cell homogenates of both strains. Presumably these two strains of A. longa differ in permeability of the plasma membrane. Further tracer and enzyme studies indicated that the Embden-Meyerhof scheme is the principal pathway of glucose catabolism; the hexose mono-phosphate shunt and the direct oxidative pathway were either not operating or quantitatively insignificant.

Notes: SYNOPSIS. By means of electron microscopy, a study has been made of the fine structure of the macrogametocytes, microgametocytes and oocysts of Eimeria perforans from the intestine of the wild rabbit (Oryctolagus cuniculus). The parasites lie in a vacuole within the host cell. The surface of the gametocytes is not plain, but displays irregular protrusions. A large intranuclear body can be detected within the macrogametocytes. Similar structures are also found within the cytoplasm. Within the latter there exists a large spread out reticulum, the channels and vesicles of which concentrate especially close to the nuclear membrane. Tubuli are seen in the numerous mitochondria, which often have a dumb-bell shape.In most of the gametocytes irregular, strongly osmiophilic lipid inclusions are observed, which always are surrounded by the endoplasmic reticulum. Strange folded ovoid bodies are found within the cytoplasm of the oocysts. Nothing can be told with certainty of their nature and function. Probably they represent specific storage bodies.

Notes: SYNOPSIS. Tritrichomonas foetus survived much better on extended storage at -95 than at -28d̀C following slow freezing in the presence of 1.0 M glycerol. There was no significant difference between these temperatures in survival up to 8 days, but thereafter the protozoa continued to die off slowly at -28d̀, whereas their numbers remained essentially constant at -95d̀ for 128 to 256 days. The trichomonads' motility was much better after storage at -95d̀ than after storage at -28d̀, and fresh cultures could be initiated from the former much more readily.Other constituents of the suspending medium besides glycerol affect the survival of the protozoa upon freezing. Survival was much better when the protozoa were frozen in the original Diamond's trypticase-yeast extract-maltose-cysteine-ascorbic acid-serum medium in which they had been grown than when they were frozen in physiological salt solution or in fresh Diamond's medium. There was no significant difference between survival in the latter two suspending media. The speed and time of centrifugation needed to remove the trichomonads from the medium in which they had been grown had no effect on their survival upon subsequent freezing. Presumably some product or products of the trichomonads' metabolism have an additional protective action which supplements that of glycerol.When frozen in the original Diamond's medium in which they had been grown plus 1.0 M glycerol, an average of 15% of the trichomonads were alive after 128 days' storage at -28d̀ and an average of 38% were alive at -95d̀C. When frozen in physiological salt solution plus 1.0 M glycerol, these percentages were 8% and 12% respectively.

Notes: SYNOPSIS. When Spathidium spathula was exposed to X-ray doses ranging from 1–25 kr this animal was found to be more radiosensitive than any ciliate previously reported. A dose of 1 kr is sufficient to increase the time of the first generation from 5 to about 5 1/2 hours. A dose of 4 kr is enough to approximately double the generation time. Bacteria in the medium during irradiation do not protect the ciliate against injury. Animals irradiated as dry cysts are only slightly more resistant than vegetative forms, requiring 10 kr to double the generation time. One day after exposure, irradiated lines are uniformly poor in growth rate (0–2 daily divisions), but later a bimodal response is noticed, some lines remaining poor and others recovering. Within 24 hours after treatment, a number of irradiated animals show structural abnormalities and are greatly increased in size. The experiments have not determined the reason for the high sensitivity of Spathidium but have made certain alternatives unlikely.

Notes: SYNOPSIS. When Tetrahymena suspended in water were given increasing doses of radiation, oxygen consumption decreased with increasing dose, reaching 60–90% at 600,000 r. Cells irradiated in 0.07 M phosphate buffer, pH 7.4, showed no significant decrease in oxygen consumption even at 600,000 r. The decrease in respiration observed on irradiation of Tetrahymena pyriformis W in water with 300,000 r of X-radiation was prevented by addition of pyruvate or acetate during or immediately after irradiation. Pyruvate stimulated the respiration of the X-irradiated cells, particularly at 10 and 60 min post-irradiation.Lactate markedly stimulated the respiration of control suspensions of Tetrahymena cells and oxidation of lactate by cells irradiated with 300,000 r was increased by 20 to 100%, depending on the concentration of lactate and the time after irradiation. Pyruvate was considerably more effective than lactate in increasing O2 uptake of X-irradiated cells, particularly at 10 min post-irradiation. Thioctic acid affected neither the respiration of control or X-irradiated Tetrahymena nor the oxidation of pyruvate.The growth lag of Tetrahymena increased proportionately with increasing radiation dose; no cells survived 600,000 r. The presence of metabolites during irradiation did not affect the lag period or subsequent growth rates. The effects observed were discussed in terms of an alteration of the permeability of Tetrahymena after irradiation.

Notes: SYNOPSIS. A full account of the nuclear changes during binary fission and conjugation in a local race of Blepharisma is presented in this paper. The macronucleus consists of 2 nodes connected by a strand. Number of micronuclei varies from 6 to 18. During binary fission, condensation of macronucleus is followed by elongation and thinning of the middle region which finally breaks. Daughter nuclei later attain the typical vegetative form. Notably, during binary fission some micronuclei appear to complete their mitoses by the time the macronucleus attains the condensed form, while others lag behind and exhibit practically every stage of mitosis.During conjugation, from 6 to 10 micronuclei undergo the first pregamic division, the same number through the second division, and two products of the second division take part in the third division. The rest degenerate. Division products of the nuclei in the paraoral region take part in synkaryon formation. The synkaryon undergoes either 2 or 3 divisions. In the former case, of the 4 products, 2 become the macronuclear anlagen, one the micronucleus and the fourth degenerates. In the latter case, of the 8 products, 3 to 4 become the macronuclear anlagen and the rest become micronuclei. Chromatin elimination has been observed during the division of the macronuclear anlage, followed by an extra metagamic fission of the cell.Comparison with two other races from India and an American race indicates considerable diversity in the structure and behaviour of the nuclear apparatus in different races of Blepharisma undulans.

Notes: SYNOPSIS. Females of Heterakis gallinae were separated on the basis of their capacity to transmit the protozoan parasite Histomonas meleagridis. Sections of worms, capable of transmitting the protozoon, revealed the organism in both males and females as well as in the eggs. Infected male worms contained histomonads in the gut wall and the wall and lumen of the reproductive system. Female worms infected with H. meleagridis showed the organisms throughout the reproductive system. Histomonads, found in uterine eggs possessing shells, had a larger nucleus and reduced cytoplasm.Because of the presence of the protozoon among the sperm in the male reproductive system, it is believed the organism can be transmitted to female worms through copulation. The cycle in the worms also supports the assumption that H. meleagridis was originally a parasite of the worm.

Notes: SYNOPSIS. Speculations regarding the mode of transmission of monocystid parasites of earthworms over a period of more than 100 years have never been tested experimentally under controlled conditions. In order to do so a stock of infectionfree Eisenia foetida (Sav.) was raised from cocoons and experimental infections were induced in this host using sporocysts of the gregarine parasites Apolocystis elongata Phillips & Mackinnon 1946, and Nematocystis elmassiani (Hesse, 1909). Experimental infections were obtained by feeding to uninfected worms sporocysts obtained directly from infected host worms. This proved that the intervention of a vector is not a necessary condition of infection. Infections could not be induced by injecting sporocysts through the body wall into the body cavity. Infections are thus probably acquired in nature by the ingestion of sporocysts. Sporocysts do not leave the body of the host by being passed from coelom to lumen of the gut, nor do they pass directly to the exterior through apertures of the body wall. There was no evidence of parasitic autotomy. It is therefore concluded that death and decay of the host is the normal method of dissemination of sporocysts. Sporocysts were not infective after drying in air for three weeks. Other sporocysts lost potency after storage in moist conditions for several months. Infections involving the organisms specified were sporadic and unpredictable; modifying factors, such as variations in host susceptibility and latency in infection, appeared to be operating.

Notes: SYNOPSIS. By an automatic electronic technique—the Flying Spot Particle Resolver—the effect of a wide range of concentrations of vitamin B12 on the size and growth of the B12-dependent Euglena gracilis was studied. Rate of cell growth was directly proportional, and cell size inversely proportional, to B12 concentration. Gross B12 depletion resulted in gigantism and prolongation of generation times.

Notes: SYNOPSIS. Previously, the reproduction-inhibiting effects of ablastin could only be shown in vivo. The present report describes techniques for the in vitro demonstration and titration of this antibody. With a medium composed of Hanks' balanced salt solution, rat serum, lactalbumin hydrolysate, yeast extract and rat blood lysate, blood stream forms of Trypanosoma lewisi can be grown for approximately 24 hours at 37d̀C. Starting with the medium containing normal rat serum and inoculated with adult (inhibited) trypanosomes from infected rat blood, 50% or more of the parasites are in various stages of division after incubation overnight. Under similar conditions with ablastic rat serum, the parasites do not reproduce but remain as adults. If the medium is inoculated with reproducing trypanosomes from the blood, parasites in the presence of normal serum continue to reproduce, whereas those exposed to ablastin are almost completely converted to adult, non-reproducing forms. Similar results are obtained when the immune sera used are first adsorbed with living parasites to remove all trypanocidal antibodies. Ablastic serum inactivated at 56d̀C for 20 minutes does not lose its inhibitory activity indicating that ablastin is not complement dependent, and parasites grown on media at room temperature are not affected by the antibody suggesting that basic antigenic differences exist between blood stream forms at 37d̀C and culture forms at room temperature. Studies of the conversion of blood stream forms to culture forms indicate that the critical temperature range for the conversion lies between 28d̀ and 30d̀C. The significance of these results is discussed, and possible applications of the techniques described to studies of the mechanism of ablastic action are considered.

Notes: SYNOPSIS. Normal swimming behavior of Paramecium multimicronucleatum in an essential mineral element solution with 0.0002 M calcium changed into continuous avoidance reactions upon replacement by equimolar strontium; equimolar barium produced a less pronounced similar effect. In equivalent pure SrCl2 and BaCl2 solutions, avoidance reactions were less frequent than in the balanced solutions. P. multimicronucleatum inoculated into autoclaved calcium- or strontium-containing cultures of the alga Protosiphon botryoides (ultimate food source) multiplied greatly and essentially equally, but died in barium. Accelerated avoidance reaction rates were observed in strontium up to and at 32 days after inoculation.

Notes: SYNOPSIS. A previous study of parasites from relapsed P. berghei infections of mice indicated that the behavior of the relapse parasites differed from that of their parent strain. Experiments have been performed comparing the behavior of relapse and parent parasite infections in mice under two sets of conditions. In one group of experiments the behavior after antimalarial treatments, designed to result in chronic or latent states of infection, was tested. In all, 8 relapse strains were tested against their parent strains in six different experiments which employed 536 mice. It was observed that the mice infected with the relapse strains had a statistically significant greater mortality after the treatments than did mice infected with parent strains. This difference was observed regardless of how the relapse strain had been previously treated, or of what treatment was used in the experiments. In a second group of experiments, the behavior of infections with relapse and parent parasites was compared in normal mice. Five relapse strains were compared to their parent strains in a total of 5 experiments using 356 mice. It was observed that the mean survival time of mice infected with relapse strains was significantly greater than that of mice infected with the parent strains. It is not known whether this apparent difference in the behavior of relapse and parent parasites is related to the mechanism for relapse of P. berghei infections, or is merely a characteristic of parasites that had survived in an immune host.

Notes: SYNOPSIS. Thymidylate synthetase (E.C.2.1.1.45) has been demonstrated in unsporulated oocysts of Eimeria tenella. The properties of this enzyme have also been investigated in Tetrahymena pyriformis, as a protozoan model, and 7-day-old chick embryo, as a host model. The enzymes from E. tenella and chick embryo were inhibited by all concentrations of MnCl2 and MgCl2 tested. Tetrahymena pyriformis thymidylate synthetase was stimulated by low concentrations of both these cations but was inhibited by high concentrations. Subsequent data refer to chick embryo, E. tenella and T. pyriformis respectively: the apparent Km was 5.89 μM, 5.94 μM, and 0.53 M for the substrate dUMP: and 5.13 μM, 1.10 μM and 4.65 μM, respectively for the cofactor N5N10-methylenetetrahydrofolate. The pH optimum for the enzyme from both chick embryo and T. pyriformis was 8.0, with Tris-HCl buffer; activity of E. tenella thymidylate synthetase was still increasing at pH 8.2. The E. tenella enzyme was found to have a molecular weight of 4.6–4.9 × 105 daltons. The effects of nucleotides, inhibitors, and the omission of assay components on each enzyme are presented. Thymidylate synthetase from E. tenella is not greatly different from that of chick embryo, but does not resemble the enzyme from T. pyriformis. A case for using thymidylate synthetase as a chemotherapeutic target in the treatment of Eimeria infections remains. Indeed Eimeria may be considered as a model for infections caused by other protozoan parasites, such as Toxoplasma and Plasmodium, provided that suitable inhibitors can be found that are not toxic to the host.

Notes: SYNOPSIS. A protocol based on density differences between starved and fed cells and employing density gradient centrifugation has been devised to facilitate the isolation of auxotrophic mutants of cell lines derived from Tetrahymena thermophila strain B1868. First, a mass phenotype screening procedure was established whereby true auxotrophic mutants and slow-growing wild-type cells such as strain C* could readily be distinguished. Second, simulation experiments were performed in which wild-type cells starved first in non-nutritive buffer, then suspended in a defined medium lacking a single essential amino acid became significantly denser than the same cells when starved, then suspended in a complete defined medium. Finally, using the same protocol, a reconstruction experiment was carried out which resulted in effective separation of wild-type cells from cells of a tyrosine auxotroph. The overall procedure resulted in a 9-fold increase in the relative frequency of auxotrophic cells, while the density gradient centrifugation alone provided a 400-fold enrichment.

Notes: SYNOPSIS. Infectivity of Plasmodium gallinaceum (Brumpt) sporozoites isolated from midguts and salivary glands of experimentally infected Aedes fluviatilis (Lutz) was studied. The 2 populations were compared at 7, 8, and 9 days postisolation from mosquitoes, which were maintained at 27 C ± 1C and ∼75% relative humidity. Infectivity of the parasites was evaluated by the length of the prepatent period of the infection in 2-week-old chicks inoculated intramuscularly. Infection was caused by 7-day-old sporozoites from salivary glands, but not from midguts. Older sporozoites induced infection in all the inoculated chicks. The results suggested a somewhat higher infectivity of the 8- and 9-day salivary-gland parasites than of the oocyst sporozoites. However, unlike sporozoites from mammalian malaria, oocyst sporozoites from avian malaria were highly infective at this age.

Notes: SYNOPSIS. Sorogena stoianovitchae Bradbury & Olive, an epiphytic ciliate found in various parts of the world, has a trophic stage that feeds on members of the ciliate genus Colpoda. When grown in the presence of the food ciliate, it multiplies rapidly. When the cells become abundant they aggregate at the water surface on inserted plant fragments or floating pollen grains, the sides of culture dishes, or on floating films such as those deposited by bacteria or pollen grains. an aggregate mounds up and becomes ensheathed above the water level, after which the mass of cells called a sorogen rises aerially at the apex of a stalk deposited at its base. the tapering, noncellular stalk consists of a conspicuously furrowed sheath that encloses a mucilaginous matrix. At completion of stalk development the cells of the sorogen become encysted. the sorocysts are commonly discharged by fracturing of the drying sorus. Alternating light and dark conditions are required for sorocarp development.

Notes: SYNOPSIS. the antigenic types in populations of metacyclic trypanosomes of Trypanosoma brucei isolated from Glossina morsitans head-salivary gland trypanosome cultures and bloodstream forms in the early parasitemias produced from whole culture supernatant fluids containing metacyclic forms, were analyzed by the indirect fluorescent antibody test using clone-specific antisera. Metacyclic trypanosomes in cultures initiated with cloned bloodstream forms were heterogeneous with respect to their variable antigenic type (VAT). Trypanosomes comprising early parasitemias in immunosuppressed mice infected with metacyclics produced in cultures also had a range of VATs. Three of the VATs detected in the early parasitemias in mice have also been identified by other investigators in tsetse fly-transmitted populations of the same stock.

Notes: SYNOPSIS. A simple method is described for plating and cloning ciliates and other protozoa, based on a principle differing from that traditionally used for plating and cloning bacteria and other microorganisms. This procedure, referred to as the silicone-oil-plating-procedure (SOPP), involves vortexing small volumes of culture medium containing protozoa with larger volumes of a non-toxic silicone oil and plating the resulting unstable emulsion in small plastic petri plates. Discrete microdroplets of culture medium form containing protozoa entrapped and immobilized between the hydrophobic surfaces of the plastic petri dish and the oil. Protozoa, isolated by this method grow, divide, and multiply to form clones. the procedure may be used for plating and cloning protozoa in bacterized and axenic culture. Variations of the basic method may be applied to isolating protozoa from the wild, washing protozoa to remove microorganisms, screening for potential mutants, and for replica plating.

Notes: SYNOPSIS. Cyclic nucleotide phosphodiesterase [EC 3.1.4.17] was examined in Tetrahymena pyriformis strain NT-1. Enzymic activity was associated with the soluble and the particulate fractions, whereas most of the cyclic GMP phosphodiesterase activity was localized in the soluble fraction: the activities were optimal at pH 8.0–9.0. Although very low activities were detected in the absence of divalent cations, they were significantly increased by the addition of either Mg2+ or Mn2-. A kinetic analysis of the properties of the enzymes yielded 2 apparent KIII values ranging in concentration from 0.5 to 50 μM and from 0.1 to 62 μ M for cyclic AMP and GMP. respectively. A Ca2+-dependent activating factor for cyclic nucleotide phosphodiesterase was extracted from Tetrahymena cells, but this factor did not stimulate guanylate cyclase [EC 4.6.1.2] activity in this organism. On the other hand, Tetrahymena also contained a protein activator which stimulated guanylate cyclase in the presence of Ca2+, although this activator did not stimulate the phosphodiesterase. the results suggested that Tetrahymena might contain 2 types of Ca2+-dependent activators, one specific for phosphodiesterase and the other for guanylate cyclase.

Notes: SYNOPSIS. the cell size of Didinium nasutum was found to be dependent on the size of the Paramecium species available as prey. Didinium feeding on P. tetraurelia averaged 5.6 × 105μm3. the cell volume of Didinium increased with increasing prey size for the 5 prey species tested, to 9.1 × 105μm3 for Didinium feeding on P. caudatum. Didinium nearing a cell division ranged in size from 8.6 × 105μm3 on P. tetraurelia to 12.9 × 105μm3 on P. caudatum. the range in cell volume is such that Didinium feeding on P. caudatum are larger than the size at which Didinium divide when feeding on P. tetraurelia. This morphologic plasticity in cell volume allows Didinium to exploit a wide size range of Paramecium species as prey. It is proposed that the size of a Didinium may have profound effects on its ability to encounter and capture prey of different sizes.

Notes: RESUME. Chacun des 45–80 organelles adoraux de Bursaria truncatella O. F. Müller est constitué de 3 rangées de cinétosomes et l'aire buccale droite est couverte de nombreuses doubles rangées de cinétosomes. La stomatogenèse débute par la désorganisation et la résorption des organelles buccaux postérieurs. Puis, il y a désorganisation des rangées parorales de cinétosomes et multiplication des cinétosomes sur l'aire orale droite, en měme temps que sont rompues, selon une ligne oblique, un certain nombre de cinéties somatiques. La prolifération des cinétosomes aux extrémités des cinéties. de part et d'autre de la ligne de rupture, aboutit, d'une part, à la formation d'un champ anarchique qui est le primordium oral droit de l'opisthe, d'autre part, à la formation de nombreux doublets qui constituent chacun le primordium de chaque organelle adoral. Après la séparation des tomites, les cinétosomes de l'aire droite s'ordonnent en doubles rangées et les organelles adoraux se complètent par addition d'une 3ème rangée de cinétosomes. Les cinétosomes somatiques sont jumelés, reliés par 2 desmoses. Les fibres transverses postérieures et les fibres postciliaires forment de longs rubans de microtubules dirigés vers l'arrière et juxtaposés dans les crětes intercinétiennes. Les doubles rangées droites de cinétosomes buccaux sont assimilables à des stichodyades. Les organelles des cinétosomes adoraux portent des rideaux de fibres postciliaires convergents ou divergents. La rangée postérieure de chaque organelle est non ciliée. Par son type de stomatogenèse, par sa structure corticale, par l'ultrastructure des organelles adoraux, Bursaria appartient aux Colpodidea, ce qui suggère des remarques de plusieurs types.SYNOPSIS. In Bursaria truncatella O. F. Müller, each of the 45–80 adoral organelles is composed of 3 rows of kinetosomes, and the right buccal area is covered by many double rows of kinetosomes. Stomatogenesis begins by disorganization and disappearance of the posterior buccal organelles. Next, there is disorganization of the paroral rows of kinetosomes and multiplication of kinetosomes in the right oral area; at the same time, some somatic kineties are disrupted along an oblique line. Multiplication of kinetosomes at the extremities of the kineties, on both sides of the disruption, leads to the formation of an anarchic field which is the right oral primordium of the opisthe and the formation of doublets each of which constitutes an adoral organelle. After the separation of the tomites. the kinetosomes in the right buccal area position themselves, and the adoral organelles are completed by the addition of a 3rd row of kinetosomes. Somatic kineties are formed by successive pairs of ciliated kinetosomes united by 2 desmoses. the long posterior transverse ribbons and the postciliary ribbons extend posteriad, overlapping in the pellicular ridges. Oral rows of kinetosomes on the right can be compared with stichodyads. the adoral kinetosomes have convergent or divergent postciliary ribbons. the posterior row of kinetosomes in each organelle is not ciliated. By the type of stomatogenesis, the cortical ultrastructure, the ultrastructure adoral of its organelles, Bursaria belongs to the Colpodidea.

Notes: SYNOPSIS Free-living marine ciliates occur in the interstitial spaces of a wide vareity of filamentous and particulate substrata, on the surfaces of planar substrata, and in the plankton. In addition, they are found in association with a wide variety of plant and animal hosts. In this paper I review the progress during the past decade in understanding the distribution of marine ciliates, with particular emphasis on the relationship between ciliate biogeography and the species problem. It is concluded that as a general rule among marine ciliates, genera and species complexes are cosmopolitan. Specific locales may support a confusing array of sibling species or subspecific morphologic variants. Because the distributional processes and breeding biology of marine ciliates are only beginning to be understood, conventional ideas that marine ciliate species are cosmopolitan may require modification.

Notes: SYNOPSIS The subkingdom Protozoa now includes over 65,000 named species, of which over half are fossil and ∼ 10,000 are parasitic. Among living species, this includes ∼ 250 parasitic and 11,300 free-living sarcodines (of which ∼ 4,600 are foraminiferids); 1.800 parasitic and 5,100 free-living flagellates: ∼ 5,600 parasitic “Sporozoa” (including Apicomplexa, Microspora, Myxospora, and Aseetospora); and ∼ 2,500 parasitic and 4,700 free-living ciliates. There are undoubtedly thousands more still unmamed. Seven phyla of PROTOZOA are accepted in this classification—SARCOMASTIGOPHORA. LABYRINTHOMORPHA, APICOMPLEXA, MICROSPORA, ASCETOSPORA, MYXOSPORA, and CILIOPHORA. Diagnoses are given for these and for all higher taxa through suborders, and representative genera of each are named. the present scheme is a considerable revision of the Society's 1964 classification, which was prepared at a time when perhaps 48,000 species had been named. It has been necessitated by the acquisition of a great deal of new taxonomic information, much of it through electron microscopy. It is hoped that the present classification incorporates most of the major changes that will be made for some time. and that it will be used for many years by both protozoologists and non-protozoologists.

Notes: SYNOPSIS Leishmania donovani amastigote-to-promastigote transformation is inhibited by homogenates of infected hamster liver and spleen. This inhibitory activity is localized in the 100,000 g pellet fraction. Tests with lysates of adherent (macrophyages) and nonadherent (lymphocytes) spleen cells indicated that the inhibitory activity resided in the lymphocytes, specifically in the 100,000 g pellet fraction.

Notes: SYNOPSIS Antibodies induced in rabbits against Paramecium multimicronucleatum syngen 2 prevent sexually reactive cells from clumping, pairing, and forming cytoplasmic fusions. A biologic assay for the detection of these antibodies (designated blocking antibodies) is described. the blocking antibodies, unlike the immobilization antibodies, are produced against breis of sexually reactive cells and nonreactive cells of 2 types, nonstarved and immature. Isolated cilia from reactive cells of either mating type are weak immunogens for blocking antibodies. No correlation between the mating type specificity (III or IV) and these antibodies has been detected. Blocking antibodies can be absorbed with living cells, of which sexually reactive ones are the most effective absorbers, while immature ones are the least effective.

Notes: SYNOPSIS A method is described for the axenic mass cultivation of Paramecium tetraurelia strains 51s and 299s. the ciliate is grown in an enriched axenic medium developed by Soldo, Godoy & van Wagtendonk. Under continuous shaking on a rotary shaker, cultures were grown in one-liter Erlenmeyer flasks with 330 ml medium yield cell densities of 32,000 cell/ml and 20,000 cells/ml for strains 299s and 51s respectively. Doubling time is considerably shorter under these conditions than in the conventional static cultures. A 20-liter airlift bioreactor is described in detail which can be used successfully to otain up to 100 g wet weight of Paramecium in a single run; in this reactor the cell density reaches 38,000 cells/ml for strain 299s. and 23,000 cells/ml for 51s. This technic should facilitate the study of minor protein components of the ciliate.

Notes: SYNOPSIS The cadmium ion (Cd2+) was accumulated by Amoeba proteus in all cellular fractions, the highest level being associated with the cytosol fraction. On gel separation of the cytosol fraction, Cd-binding protein appeared in 2 peaks: one 〉45,000 MW (peak I) and the other 12,000 MW (peak II). Added cysteine increased the total Cd2+ taken up by the cells and resulted in disproportionate increase of Cd incorporated into the Cd-binding protein of peak II. the Cd-binding protein of peak II is analogous to the low-MW, Cdbinding proteins in Anacystis nidulans, Mytilus edulis, and to the metalloprotein of some vertebrates.

Notes: SYNOPSIS. Schizonts were found in the middle and lower third of the small intestine of two calves killed 12 and 14 days after they had been inoculated with pure cultures of oocysts of Eimeria auburnensis. The schizonts ranged from 78 to 250 μ long by 78 to 150 μ wide (mean 92 by 139.9 μ). They were usually located deep in the lamina propria near the muscularis mucosae instead of in the villi where most schizonts of Eimeria bovis are found. The schizonts of E. auburnensis resembled the previously described large microgametocytes of this species but were distinguishable morphologically and by histochemical stains. The microgametocytes were much larger than previously reported; one measured 91 by 287.5 μ.

Notes: SYNOPSIS. Three new species of Blepharisma are presented: B. seshachari sp. nov., B. intermedium sp. nov., and B. tropicum sp. nov. Two sub-species of Suzuki are elevated to specific rank, B. undulans americanum to B. americanum and B. undulans japonicum Suzuki to B. japonicum Suzuki. Stein's taxa for the species Blepharisma undulans are considered to be diagnostic for that species.Important features of morphology and life-cycle of the above mentioned species are given. Discussion and position of the species Blepharisma undulans are presented and a proposal for the reorganization of the species is made. The possibility of a new key derived from a hypothesis of the phylogeny on an evolutionary basis of the genus is presented. This hypothesis has been extended to include ciliates like Stentor and Spirostomum and its implications discussed.

Notes: SYNOPSIS. A laboratory colony of Oncopeltus fasciatus was found to be infected with Leptomonas oncopelti. Inasmuch as the parasite is transmitted from parent to offspring an opportunity presented itself to study the biology and transmission of this parasite under controlled laboratory conditions. An apparatus for observing individual bugs was designed and the presence or absence of flagellates in the feces determined. Flagellates were not shed until the bugs became adults after which they appeared in every defecation. Dissection of infected bugs revealed that flagellates were not present in the rectum until adulthood. Further studies indicated that in the midgut of the insect there is a departure from binary fission to budding. The nucleus divides and one of the newly formed nuclei migrates toward a newly formed kinetoplast. Rarely there is still another kinetoplast/nucleus division. In the event the new axoneme grows within the cytoplasmic sheath of the parent flagellum, smaller organisms produced by unequal cytokinesis remain attached. If the axoneme grows free, the smaller daughter organisms become free-swimming. Passage into the rectum of the adult bugs causes a rounding up of all parasites although the leishmaniform organisms continue to divide. It is presumed that infection of clean bugs is accomplished by the ingestion of leishmaniform organisms through a common water source. The reason for the presence of flagellates in the rectum of the adult but not in the nymphal insect and the mechanism responsible for the change from binary to unequal fission are not known.

Notes: SYNOPSIS. Examination of intestinal contents and feces of Wisconsin red squirrels (Tamiasciurus hudsonicus) revealed two coccidia, Eimeria tamiasciuri and E. toddi n.sp. In the Eastern chipmunk (Tamias striatus), two species, E. vilasi n.sp. and E. wisconsinensis n.sp. occurred. In the northern flying squirrel (Glaucomys sabrinus), a coccidium resembling E. sciurorum was found.

Notes: SYNOPSIS. A large heterotrich ciliate (Family: Bursariidae) found in a papyrus swamp in Uganda was used for oxygen tension experiments by Beadle & Nilsson, 1959, under the name of Bursaria sp. This organism has now been identified as Neobursaridium gigas Balech. The morphology of the organism was studied in living and stained specimens, especially with the silver impregnation technique, and the present findings are compared to those of Balech.

Notes: SYNOPSIS. An electron microscope study of thin sections of Babesia rodhaini parasitizing mouse erythrocytes showed that this intracellular parasite feeds on its host cell in the same way as do malarial parasites. Large portions of erythrocyte cytoplasm are engulfed by invaginations of the plasma membrane leading to the formation of food vacuoles. The digestion of hemoglobin seems to be complete in Babesia since no pigment (hemozoin) could be detected either in the food vacuoles or in the cytoplasm.The fine structure of Babesia rodhaini is very similar to that of Plasmodium berghei. In both, typical mitochondria are lacking. Instead a structure was found composed of concentric membranes which it is assumed might perform mitochondrial functions. As in Plasmodium berghei a double-membraned vacuole with a matrix of low density is present in almost all sections. The endoplasmic reticulum is represented by small vesicles; the ground-substance is filled with Palade's small particles. The nucleus is large and surrounded by two membranes. Babesia reproduces by budding and binary fission. Reproduction is not easy to identify since the parasite forms large pseudopods resembling stages in division.

Notes: SYNOPSIS. The effect of 10 detergents on the inhibitory activity of neomycin was tested against Ochromonas danica. All 4 anionic detergents used were synergistic with neomycin. Evidence is presented that the synergisms were due to the effect of the detergents on the cell membrane. In the presence of Tergitol 7, O. danica is more susceptible to changes in osmotic pressure by increasing concentrations of KCl.

Notes: SYNOPSIS. All zoologists are affected by provisions in the very recently published International Code of Zoological Nomenclature, the first revised edition of these important rules to appear in over 50 years. Common nomenclatural practices, often malpractices, of protozoologists and parasitologists who work primarily in taxonomic fields are revealed and discussed in light of recommendations and mandatory regulations to be found in the new Code. Some errors have been due solely to carelessness; others have involved misinterpretations of various directives; still others have involved cases not adequately covered by the old Règles. Certain mistakes of the past cannot be changed; but others are to be rectified upon discovery, according to mandates in articles of the new Code. Practical applications of the rules of nomenclature are stressed, and examples are taken from actual situations found to exist throughout all major taxa of the phylum Protozoa.Because of the value of such discussion in both new and revisory work in protozoan systematics, the following major topics are given special consideration: matters of orthography, the original spelling of names and their justified or unjustified emendation; authorships and dates of names, who is responsible and when, and how such data are properly cited; mandatory dates in the new Code, and their effect on both already established names and names not yet proposed; the principles of priority and conservation or continuity, and how the rules attempt to satisfy proponents of both of these diametrically opposed “laws”; the concepts of synonymy and homonymy, and proper methods of treating names which have become involved in such situations; family-group names, and the several special nomenclatural problems they present to protozoan taxonomists; the major problem of types, and the peculiar position of protozoologists with regard to the type concept, especially type-specimens for categories in the species-group; miscellaneous considerations, several unrelated but significant topics not appropriate for inclusion in preceding sections of the paper.

Notes: SYNOPSIS. In a chemically-defined medium Tetrahymena setifera HZ-1 required a sterol, an alcohol, eleven amino acids, a purine, a pyrimidine, and six B-complex vitamins. The sterol requirement was met by a variety of 3β-OH, C27-C29 sterols including cholesterol and stigmasterol, but not by precursors of cholesterol which precede desmosterol or Δ7-cholestenol. Some combinations of long-chain fatty acids with a synthetic dipalmitoyl phosphorylethanolamine partly substituted for sterol. Ethyl and methyl alcohols (but not a variety of other alcohols and organic acids) satisfied the alcohol requirement.

Notes: SYNOPSIS. Trichomonas vaginalis grown aerobically slowly metabolizes glucose-U-C14 and succinate-2,3-C14 to CO2 and to amino acids which are then incorporated into protein. Analysis of protein hydrolysates from cells grown on glucose-U-C14 reveals radioactivity in 15 amino acids. Although the data do not permit a direct interpretation and analysis of the sequence of amino acid biosynthesis, it seems likely that a tricarboxylic acid cycle operates. Attempts to prepare cell-free systems carrying out cycle reactions have proved unsuccessful.

Notes: SYNOPSIS. One hundred eighty-eight fresh-water samples from France (30), Italy (42), Austria (12), Germany (1), Switzerland (35), Holland (20), England (26), and Sweden (22) were examined for T. pyriformis. The habitats included rivers, mountain streams, lakes, ponds, irrigation ditches, roadside ditches, running and dead water canals, and fountains. The temperature ranged from 10° to 30°C, locations from 42° to 68° North latitude, with elevations from sea level to approximately 3,000 feet. Of the samples taken 28 contained T. pyriformis from which 411 clones were established in axenic media. Thirty-three additional samples contained ciliates other than T. pyriformis. All clones grew well in 1% proteose peptone; when screened for their nutritional requirements a few failed to grow on the completely defined medium and several others survived eight sub-inoculations without thiamine.The nuclear and sexual activity pattern followed that of previous collections. In distribution, variety 6 was found in Italy, variety 3 in Austria, and variety 4 in England. A new variety (variety 10) with two mating types was isolated from four different habitats in England. Several strains from Italy, France, Holland, and England constitute one group which mated among themselves, but only a few of which reacted with mating type I of variety 6 from America. This demonstrates the close affinities of the European and American strains, yet shows the possible evolution of a new variety (species).

Notes: SYNOPSIS. The steroid requirement of a Trypticase-requiring strain, SLT, of Trichomonas gallinae on the Shorb-Lund synthetic basal medium minus ribonucleic acid was fulfilled by cholesterol, 7-dehydrocholesterol(provitamin D3), cholestanol (dihydrocholesterol), and to a lesser degree, by ergosterol (provitamin D2), β-sitosterol and zymosterol acetate. Cholestanol was inhibitory at certain concentrations. All steroids were subsequently checked by gas chromatography. Only cholesterol and zymosterol acetate showed one peak, the others containing 2 to 4 peaks. Repurified cholestanol was active and showed no inhibition of growth, but repurified β-sitosterol was inactive. The following were also inactive: diethylstilbestrol, estrone, dehydroepiandrosterone, progesterone, testosterone, desoxycorticosterone acetate, cortisone acetate, compound A, 5,6-dihydro-ergosterol, dihydroergosterol acetate, stigmasterol, sodium taurocholate, cholic acid, methyl desoxycholate, desoxycholic acid and methyl cholate. Precursors in cholesterol synthesis, acetate, mevalonic acid, mevalonic acid plus a liver extract and squalene were inactive in replacing cholesterol, although acetate is required for growth in the presence of cholesterol. Crystalline fat soluble vitamins, carotene, calciferol (vitamin D2), activated 7-dehydrocholesterol (vitamin D3), menadione (vitamin K3) and tocopheryl acetate (vitamin E acetate) were inactive in replacing cholesterol. Trichomonas species PN from the pig's nose utilized cholesterol, 7-dehydrocholesterol and the impure β-sitosterol sample in a modified SL medium with acetate omitted. A reinvestigation of steroid requirements seems to be indicated when a completely synthetic medium and pure steroids are available.

Notes: SYNOPSIS. Protoplasm transferred to a multinucleated amoeba is well tolerated if the donor and recipient portions are from the same amoeba, from different amoebae of a single clone, from different amoebae of the same species found in a given location, or from different amoebae of the same species found in widely separated geographic areas. On the other hand, protoplasm is not tolerated and death invariably follows interspecific or intergeneric microtransfers between any two of the three species of amoebae used in this study.Attempts to overcome the tolerance block in heterologous transfers by the use of x-radiation failed although a significant extension of the mean survival time was obtained. The latter can also be interpreted as a limited therapeutic effect of nonirradiated, heterologous donor protoplasm on supralethally irradiated recipients.

Notes: SYNOPSIS. Cultures of Prymnesium parvum subjected to constant illumination failed to produce ichthyotoxin. On the other hand cultures subjected to alternate periods of light and darkness showed a gradually rising ichthyotoxic activity during the dark period reaching a maximum after about 7 hours.

Notes: SYNOPSIS. A new species, Crithidia rileyi (Trypanosomatidae), is described from Tabanus epistates. The most characteristic feature of the flagellate is the frequent occurrence of a broad sucker-like posterior end. In contrast to other species of Crithidia it multiplied very slowly in the first few weeks after isolation. A review of the literature shows that trypanosomatids of two generic types, Crithidia and Blastocrithidia, have been described from flies of the family Tabanidae, although they have nearly always been regarded as stages of the same organism. Some or all of those of the Blastocrithidia type are stages of Trypanosoma theileri. The Crithidia are probably not stages in the development of any trypanosome.

Notes: SYNOPSIS. Gromia oviformis from New Zealand has been examined in thin sections after fixing in osmium tetroxide and embedding in methacrylate. The structure of the protein and acid mucopolysaccharide shell and the canals which pass radially through it are described. Multiple honeycomb membranes located between the shell and the cytoplasm, as well as mitochondria and “Golgi bodies,” none of which has hitherto been seen in light microscope studies, are discussed and illustrated. The previously known stercomata and xanthosomes are considered. Though still regarded as enigmatic cytoplasmic inclusions, observations suggest that they are waste products retained within the animal throughout life.

Notes: SYNOPSIS. An agar overlay technique developed to investigate the effect of various substances on growth of a selected group of protozoa is described. Test systems employing Crithidia fasciculata (culex), Ochromonas malhamensis, Tetrahymena pyriformis W and Trichomonas foetus were devised to evaluate the effect of known compounds and to assay the activity of antibiotic fermentation beers. This disc-plate method can be summarized as follows: preparation of a foundation layer; overlaying of a mixture of agar, nutrients, protective antibiotics, and inoculum; placing of paper discs with the test material on the overlay followed by incubation and subsequent observation of the plates for zones of inhibition of growth. The concentration of agar in the overlay sufficient to restrict movement of the protozoa without inhibiting their growth, nutrients required for adequate growth and continued viability of the inoculum, and the concentration of penicillin and streptomycin which will not inhibit the assay protozoa but beeffective against the microorganisms in the fermentation beers are discussed. This technique offers a method not only for primary screening for cytotoxic substances but also one which may be adaptable to other studies of protozoa.

Notes: SYNOPSIS. Acid phosphatase activity was studied in total mounts and sections of agnotobiotic Paramecium multimicronucleatum by the alpha-naphthyl phosphate-hexazotized rosanilin method. Timing was achieved by India ink marking of food vacuoles. Enzyme activity is present in small endoplasmic granules and in the greatest part of food vacuoles. Following an inactive stage (stage I) of an average length of 5 min the activity appears at the periphery of the vacuole, in most cases in the form of granules (stage II). A high activity level (stage III) is attained within 1 1/2 min and maintained for the most part of the vacuolar cycle. The activity disappears only in the latest vacuoles before egestion (stage IV). The appearance of activity is not concurrent with but succeeding to the maximum of vacuolar acidity as ascertained by feeding Congo red stained killed yeast cells. On the basis of these results the food vacuoles may be looked upon as belonging to the lysosomes sensu lato.

Notes: SYNOPSIS. In Oxytricha fallax, extrusion of all the excretory crystals and some refractile bodies occurs when food is with held. The precystic animals thus resulting become transparent and undergo dedifferentiation of organelles. First the undulating membrane and cytostome are entirely resorbed, producing astomial forms. Then the adoral membranelles vanish gradually, accompanying which the somatic ciliature such as the cirri comes to be resorbed. Occasionally, early precystic animals retaining the intact feeding organelles reorganize spontaneously (physiological regeneration), but they become irreversibly encystable by high-temperature treatment; for hastening resorption of feeding organelles accelerates encystment.When transparent animals at various precystic stages are cut in half the posterior fragment (PF) always encysts faster than the anterior one (AF) from the same parent, except that some of the AFs from early stages reorganize into normal vegetative forms. The time difference for completion of encystment between an AF and PF pair is at early stages larger than at later ones. If the anterior part of AF is further excised, the time difference for encystment between AF and PF from the same source becomes smaller or indistinguishable, with the exception that some AFs from early stages regenerate instead. Further, when the anterior part of whole precystic animals is removed, the time for their encystment is reduced. Also in this case, the fragments from early stages normally regenerate.From these results, it is presumed that the adoral membranelles tend to inhibit the encystment process as they do the initiation of an oral primordium in dividers and regenerants of Stentor and other ciliates. Accordingly, when resorption of membranelles becomes extensive as precystic processes progress, their dominance over and inhibition of cyst formation may be diminishing. From the fact that regeneration of fragments from early stages into normal forms may be due to retention of the undulating membrane, its resorption is considered to be an essential prerequisite for the initiation of irreversible encystment.

Notes: SYNOPSIS. A partially purified hexokinase extracted from Euglena had the following properties: 1) pH optimum 8.0–8.5; 2) an absolute requirement for Mg++ and adenosine triphosphate with Mn++, Co++, Zn++, and Ca++ less active or inactive under the test conditions. Inosine triphosphate could not be substituted for adenosine triphosphate; 3) an apparent Km (glucose) of 5 × 10−4 M; 4) the enzyme was not inhibited by the usual sulfhydryl agents, but was inhibited 24% by adenosine diphosphate. The extract phosphorylated fructose and mannose. It was not clear whether a separate enzyme phosphorylated fructose. Relative rates of phosphorylation, as determined by labile phosphate analyses, were: glucose, 1.00; fructose, 1.19; mannose, 0.56. The partially purified extract also contained phosphohexose isomerase activity; 6-phosphogluconic acid dehydrogenase was absent. Euglena hexokinase is apparently localized in the cytoplasmic or small-particle fraction of the cell. In most respects the enzyme seems to resemble the well-characterized yeast hexokinase rather than the muscle enzyme.

Notes: SYNOPSIS. Lankesterella bufonis sp. nov., from toads, Bufo regularis Reuss, in Egypt, described herein, appears to be the first species of this genus to be found in Bufo. Sporozoites of the parasite in circulating blood are usually intracorpuscular, rarely free. Free forms, abundant in smears of internal organs, particularly from lungs, are small, slender, slightly curved bodies with the anterior end more pointed than the other. The small, oval, delicate nucleus is always situated in the posterior third of the parasite, either terminal or more usually subterminal. A single cytoplasmic vacuole is located just anterior of the nucleus and more-or-less centrally. Few darkly stained granules are sometimes seen around the vacuole. Though intracorpuscular forms retain morphological characteristics of the free forms, they frequently undergo peculiar changes: apparently flattening of host-cells during smearing and fixation results in shorter and wider, and sometimes greater width of the parasite. Multiple infection with two and rarely three Lankesterella sporozoites is not uncommon. In liver and spleen smears, certain large cells, apparently of the macrophage or endothelial types, contain forms similar to those described in the peripheral blood, but mostly shorter and broader. Various developmental stages of schizonts were also seen in the liver and lung. In liver, lung, spleen, and kidney sections, numerous young forms, possibly merozoites, were scattered among tissue cells. The present parasite differs morphologically from Lankesterella minima (Chaussat, 1850), L. monilis Labbé, (1894), and L. canadensis Fantham et al., 1942.

Notes: SYNOPSIS. Ophryoglena mugardi is an Ophryoglena of medium size characterized by its spindle-shaped and bimicronuclear theront, by its rather bulky trophont and by its physiological reproductive cycle. During starvation its physiological evolution causes a secondary and temporary encystment of the theront lasting 6 or 7 days (at 18°C). The ultimate re-encystment leads to cytolysis. Feeding induces the physiological reproductive cycle; if food is given soon after the release of the tomites, it determines a primary cycle, usually allowing eight tomites; if it is dispensed only after the secondary encystment of the hungry theront and after its excystment it causes the appearance of secondary cycles, allowing two or four tomites.When the starved theront, becoming smaller and smaller, is fed it produces trophonts which encyst. These no longer divide; each one delivers one theront only. These theronts, smaller and smaller, are able to produce the original major forms, but only after several successive alimental cycles and without any secondary encystment of the theront.

Notes: SYNOPSIS. The plastid system of “normal”Euglena gracilis strain Z was found to be extremely unstable. Under normal culture conditions in the light about 1 or 2% of the cells were found to have lost spontaneously the capacity to green on further culturing. Cells treated with streptomycin, heat or U.V. all lost the capacity to green on further culturing. Bleached cells whether appearing spontaneously or by induction with streptomycin, heat, or U.V. light, were all found to possess within them organelles which were identified as proplastids.The proplastids of some “bleached” strains were capable of synthesizing porphyrins when grown in a standard culture medium. Others synthesized porphyrins only after the addition of delta-aminolevulinic acid to their medium, while proplastids of still other strains could not synthesize porphyrins from this precursor. Normal cells when grown in total darkness were found to possess proplastids morphologically identical with those of the bleached strains. Upon exposure to light the proplastids enlarged and greened. In cells which grew under continuous light the plastid system appeared as an interconnected system of tubules. Cells maintained on a schedule of 12-hour light and 12-hour dark had plastids which were detached from each other.

Notes: SYNOPSIS. Euglena and Chlamydomonas were cultured in an organic medium in the dark and at several light intensities (15, 60 and 150 ft-c) at temperatures from 20d̀ to 35d̀C. Below 32.5d̀, growth of Euglena was independent of light. Chlamydomonas was light dependent at all temperatures where growth occurred; there was no growth in the dark, at 15 ft-c, or at temperatures above 32.5d̀. At 35d̀, growth of Euglena became inversely light dependent; the higher the illumination, the poorer the growth. Multinucleated, giant euglenas were found at 35d̀, a greater percent of abnormal cells appearing at the higher light intensities. Monsters were not observed in Chlamydomonas.To explain the growth-inhibiting, monster-inducing effect of elevated temperatures on Euglena, it is postulated that a dark-formed thermosensitive protein, essential for normal cell division, is denatured. Light may increase the effect of heat on chlorophyll and the chloroplast, possibly by being converted to intraplastidic heat through the plastid carotenoids, thus having some indirect synergistic role in the phenomenon.

Notes: SYNOPSIS. Erythromycin bleaches Euglena gracilis in a manner resembling that of streptomycin. Erythromycin-bleached substrains have been cultivated 16 months in light on erythro-mycin-free media without greening. Bleached substrains were obtained only if erythromycin was added to actively growing cultures: erythromycin did not bleach if added during the stationary phase of growth of green cultures.

Notes: SYNOPSIS. Conchophthirus curtus is a thigmotrich that occurs on the gills of Unionid clams (Elliptio complanatus in this study). The anterior third of the endoplasm, unlike the remainder, is relatively firm and without gastrioles; it contains an extensive aggregation of specialized endoplasmic granules and is therefore called granuloplasm. On the surface ventral to the granuloplasm is a thigmotactic area that bears closely set, strongly adherent cilia. There is no evidence that the granules are intracellular microorganisms; they are Feulgen-negative and do not divide, nor do they stain like bacteria. Cytochemical tests show that the granules contain neutral fat, fatty acid, phospholipid, glycogen, and mucin. The evidence indicates that the principal function of the granules and granuloplasm is the production of mucin, which is supplied to the underlying thigmotactic cilia, thereby conferring on them their adhesive properties. Thus, the granules and granuloplasm constitute a mucous organelle, and to the extent that they are osmiophilic and secretory they qualify as Golgi bodies and Golgi material, respectively. Since endoplasmic granule is a general term for any of the granules of protozoan endoplasm, it is recommended that the granules of the present study be called muciferous granules.

Notes: SYNOPSIS. This is a description of structures seen in EM preparations of the macronucleus of Paramecium aurelia, using both starved and growing organisms, the latter at timed stages after fission. The effects of treatment with RNA-ase, DNA-ase and of the “silver-Feulgen” reaction are described, and comparisons made between structures in the macronucleus and the micronucleus. The appearance of macronuclei in thin (2 μ) paraffin sections, stained by the Feulgen, azure A and pyroninmethyl green methods, and upon examination by ultra-violet micrography, is also considered. From these observations it is concluded that the macronucleus contains a large number of structures, 0.5 μ in diameter, consisting of an outer RNA-containing region and central elements containing DNA. These bodies, which are usually thought to be “nucleoli” by other workers, are now considered to be the most likely candidates for the genetic “sub-nuclei” which have been postulated from genetic work.

Notes: SYNOPSIS. The longitudinal kineties of the holotrich ciliate Trimyema marinum each bear only three widely spaced cilia; but these cilia are located at identical horizontal levels so that they comprise three helicoidal ciliary bands running around the anterior pole of the organism. The two kineties whose cilia are found most anteriorad, located in a right dorso-lateral position on the body, are exceptional, however: they are prolonged anteriorly, bearing numerous short additional cilia.These two “polykineties” wind clockwise around a kind of apical disc; their distal extremities continue to curve, dipping into a “prebuccal invagination” in which they may be said to constitute a vestibular ciliature.In spite of its rather unique characteristics, the curious anatomy of this ciliate does conform to the organizational type recognized for holotrichs of the order Trichostomatida.

Notes: SYNOPSIS. A number of small molecules including CO2, certain Krebs cycle acids and certain amino acids stimulated growth and glucose utilization of a colorless strain of Euglena gracilis in a phthalate-buffered mineral salts medium. Their major effect was to shorten markedly the lag period; some compounds also gave small increases in maximum growth rate and total cell yield. The stimulatory substances were effective in small (sparking) amounts. It is suggested that all the stimulatory substances acted similarly, by keeping the internal concentration of oxaloacetate at a sufficiently high level to allow effective operation of the terminal respiratory cycle.Glucose utilization was initiated within a pH range of 3.0 to 5.0. This range could be extended by the addition of glycine to the growth medium. Cells cultured at pH 4.5 could be adapted to growth at pH 7.0 in a phosphate-buffered mineral salts medium containing a small amount of citrate. Neither adaptation nor growth occurred at pH 7.0 in the absence of citrate. Since no evidence of citrate utilization was obtained and since EDTA replaced citrate, it is concluded that citrate was acting as a chelator.The major enzymes of the Embden-Meyerhof-Parnas and pentose phosphate pathways were demonstrated in cell-free extracts. The data presented indicate that glucose metabolism in Euglena is similar to that found in a wide variety of other cells. Indirect evidence was also obtained for the operation of a uridine linked pathway in hexose metabolism.With the exception of hexokinase, for which results were inconsistent, all the enzymes demonstrated were routinely found in the supernatant fraction of the cell-free extracts. The pH optima of a number of the enzymes were found to be between pH 7.5 and 8.6, although the cells from which the enzymes were obtained were grown in acid medium.

Notes: SYNOPSIS. Cannibal giant Blepharisma undulans americanus, NYU strain, grown in mass culture, showed increased numbers of macronuclear lobes and greater variability in their number, compared to the bacteria-fed form. Feulgen and methyl greenpyronin preparations stained both the DNA and RNA of the cannibals and of the ingested forms within the food vacuoles.During the first 3 to 6 hours of digestion, macronucleus and deoxyribonucleic acid disappear from the food vacuole; during the last 6 hours the ribonucleic acid disappears from the food vacuole; DNA and RNA are therefore presumed to be digested and absorbed.

Notes: SYNOPSIS. Production of a “vitamin B22-like compound” by Tetrahymena pyriformis mating type II, variety 1, and 3 other tetrahymenids was demonstrated. The compound produced by T. pyriformis was nutritionally active for Escherichia coli and Lactobacillus leichmannii and inert for Ochromonas malhamensis and Euglena gracilis, despite use of a variety of the standard methods for liberating bound B12. By dry-weight it is present in 1 part in 30 million. Its chromatographic and electrophoretic properties are described.

Notes: SYNOPSIS. Turnix suscitator (Galliformes), Phaenicophaeus javanicus (Cuculiformes), and two passerine birds, Dicaeum cruentatum and Pycnonotus dispar, harbor Plasmodium vaughani Novy and MacNeal in the States of Pahang, Perak and Trengganu. P. rouxi Sergent, Sergent and Catanei occurs in the same localities and in Kelantan too, in the following eight passerines: Chloropsis cyanopogon, Copsychus malabaricus, Geokichla sibirica, Malacocincla rostrata, Pitta megar-hyncha, Pomatorhinus hypoleucos, Pteruthius erythropterus and Pycnonotus goiavier. This is the first report of either parasite from Malaya, and from the hosts listed. Taxonomic criteria are discussed, and it is pointed out that Dr. A. H. Helmy Mohammed's “bow-tie” schizonts are a useful guide to the recognition of P. rouxi.

Notes: SYNOPSIS. A leptomonad flagellate found in large numbers in the latex of the asclepiad plant Pergularia extensa in East Africa has been identified as Phytomonas elmassiani (Migone). The flagellate is believed to undergo cyclical development in the lygaeid bug Oncopeltus famelicus which feeds on Pergularia. Following the ingestion of infected latex by the insect, the flagellates on reaching the midgut enter a growth phase and increase in size to form giant leptomonads. These probably then migrate to the salivary glands of the bug, possibly by boring through the gut wall, haemocoel and walls of the salivary glands. The salivary gland forms represent a multiplication phase in the life-cycle, and numerous small metacyclic forms are produced which are then injected into a plant host to initiate a new infection.Also infecting the midgut of Oncopeltus famelicus is another trypanosomatid, Blastocrithidia familiaris (Gibbs). It is characterised by the possession of an undulating membrane and the habit of budding off leishmania bodies from its anterior end.These bodies are resistant and serve in the transmission of the parasite from bug to bug.The two flagellates of Oncopeltus famelicus are readily distinguished from one another, not only on morphological grounds but also on their behaviour in vitro. B. familiaris can be cultivated on a monophasic glucose-peptone broth. It then loses its undulating membrane but retains its budding habit. Phytomonas elmassiani will not grow on this medium but when inoculated into a diphasic peptone blood-agar medium the latex forms will undergo development comparable to their insect midgut phase.The plant trypanosomatids appear to form a well-defined group bearing little similarity to the monogenetic insect flagellates of the “crithidia” type. It is suggested that the culture form Strigomonas (=Crithidia) oncopelti, allegedly parasitic in asclepiads and Oncopeltus fasciatus, might be simply an insect parasite, its original plant association being quite fortuitous.

Notes: SYNOPSIS. Ten- or 12-day cultures of Trypanosoma ranarum in Diamond's SNB-9 medium contain large numbers of slender crithidia and considerably smaller numbers of pear-shaped crithidia, but in fresh media the pear-shaped bodies constitute about 93% of the population after 5 days whereas slender forms make up about 4–5%; the remainder are leishman bodies. Succeeding days show a different cycle in that the slender forms increase to some 93% at 10–13 days and the pear bodies decrease to some 3% of the total at the same time.When washed in Krebs-Ringer phosphate buffer alone, neither morphological type shows endogenous respiration; in glucose containing buffer, O2 consumption becomes evident; pear-shaped crithidia use, on an average, 12.6 μl O2/hr/108 whereas slender forms utilize 17.7 μl O2/hr/108 parasites. Malonate at 0.01 m inhibits O2 consumption of slender forms about 61% as compared to 27% with the pear-shaped bodies. A speculative discussion of the relation between morphological types, O2 consumption in the presence of glucose, and malonate inhibition is presented.

Notes: SYNOPSIS. Short sections of deoxyribonucleic acid (DNA) in the elongated macronucleus of Euplotes eurystomus were labeled by means of a short exposure to tritiated (H3-) thymidine to follow by autoradiography the fate of the labeled DNA during amitotic reorganization of the macronucleus. During amitosis the radioactive DNA that was restricted during interphase to short sections of the nucleus is dispersed and becomes evenly distributed throughout each daughter macronucleus.Although the reorganization bands normally originate only at the ends of the macronucleus, additional bands can start at other places on the macronuclear surface. Bands of the same macronucleus originate with a high degree of synchrony. DNA synthesis is a constant feature of the rear zone of every reorganization band.

Notes: SYNOPSIS. Thin sections of Lankesterella garnhami were examined by electron microscope and the morphology of trophozoites and sporozoites is described. The envelope of the organism consists of a double membrane, which (in the presumed sporozoite) is broken at one point posteriorly by the microphyle. The anterior end possesses characteristic organelles, which include an open apical ring leading to a collar or conoid and to 30 peripheral fibrils, a paired organelle, and numerous “lankesterellonemes.” Typical mitochondria and a Golgi apparatus are found in the cytoplasm and a nucleus of heterogeneous nature.

Notes: Books reviewed in this article:Larwood, G. & Rosen, B. R., eds. 1979. Biology and Systematics of Colonial Organisms.Hellebust, Johan A. & Craigie, J. S. eds. 1978. Handbook of Phycological Methods. Physiological and Biochemical Methods.

Notes: SYNOPSIS. Surface saccharides in 2 Trichomonas vaginalis strains, the moderately pathogenic, JH34A, and the mild, JH162A, were analyzed with the aid of plant lectins. Concanavalin A (Con A), wheat germ agglutinin (WGA), soybean agglutinin (SBA), castor bean agglutinin (CBA), and lectin from the garden pea (GPA) were employed in agglutination tests and in treatment of ultrathin sections for electron microscopy according to the horseradish peroxidase-3,3′-diaminobenzidine method. With Con A and WGA, small quantitative differences were noted between the 2 strains in the results of agglutination and in the reaction-product deposits observed by electron microscopy. Distribution of the binding sites for the 2 lectins was also somewhat different in the JH34A and JH162A trichomonads. In general, the reactions with the more pathogenic strain were slightly stronger. Although the reactions with SBA and CBA lectins were weaker than those with Con A or WGA, they provided the means for qualitative differentiation between the 2 trichomonad strains. SBA alone agglutinated the JH34A strain and formed demonstrable deposits on the cell surfaces. On the other hand, only CBA reacted with JH162A flagellates. The garden pea lectin failed to bind to the surface of either strain. On the basis of results obtained with the control preparations incubated in the presence of specific inhibitors, it was concluded that both strains had α-methyl-D-mannoside and/or α-methyl-D-mannoside-like as well as N-acetyl-D-glucosamine residues on their surfaces. In addition, JH34A strain had D-lactose-containing residues while JH162A trichomonads had residues with D-galactose. Neither strain appeared to possess residues containing N-acetyl-D-galactosamine.

Notes: SYNOPSIS. Centrifugation for 30–40 seconds at 8,000 g has been used to render monopodial specimens of the large free-living ameba. Chaos carolinensis. These monopodial amebae exhibit obvious torsional movements in the tail. In many cases the posterior ectoplasm assumes the form of a screw with helical ridges forming in place of the more common straight dorsal fins. This finding prompted a re-examination of normal polypodial C. carolinensis, and a majority of these were found also to exhibit torsional movement in the tail and in retracting pseudopodia. These movements suggest that the cytoskeleton of Chaos may have a helical component in its organization.

Notes: Books reviewed in this article:Levandowsky, M. & Hutner, S. H., eds. Biochemistry and Physiology of Protozoa.Raymont, John E. G. with J. D. Burton & K. R. Dyer. 1980. Plankton and Productivity in the Oceans. Vol. I. Phytoplankton.

Notes: Book reviewed in this article:Maramorosch, Karl & Hirumi, Hiroyuki, eds. 1979. Practical Tissue Culture Applications.

Notes: SYNOPSIS. Cysteine and ascorbic acid were previously shown to be required by Entamoeba histolytica trophozoites for attachment to glass, elongation, and ameboid movement as well as for short-term (12–24 h) survival in a balanced salt solution containing bovine serum albumin and a vitamin solution (Maintenance Medium 1). If the only function of cysteine and ascorbate was to decrease the redox potential, other reducing agents should be effective. However, the requirement for cysteine in the presence of ascorbic acid was highly specific. Equally effective were D- and L-cysteine; however, of many other compounds tested, only thioglycolic acid, ascorbic acid, or L-cystine (in decreasing order) were somewhat active. Under N2 atmosphere, cysteine and ascorbic acid were still required, although their concentrations could be halved. The ability to attach in the maintenance medium was irreversibly lost after only 5 min of cysteine-ascorbic acid deprivation; however, there was no decrease in viability when the amebae were transferred to growth medium within 30 min. Cysteine thiol groups in the medium were oxidized rapidly regardless of the concentration of ascorbic acid or the presence of amebae; however, ascorbic acid prolonged attachment of amebae.

Notes: SYNOPSIS. Low concentrations of chlorpromazine (∼0.01 mM) inhibit growth and nucleic acid synthesis in the ciliate Tetrahymena pyriformis. Brief exposure of the cells to, e.g. 0.018 mM chlorpromazine, had very little effect on 14CO2 production or on label incorporation into glycogen from [1-14C]glucetate, [6–14C]glucose, or [1-14C]leucine, but 17-h exposure of stationary phase cultures to this drug caused marked alterations in metabolism, including an almost complete loss of ability to decarboxylate L-[1-14C]leucine and L-[1-14C]tyrosine. It was shown that loss of ability to decarboxylate these amino acids results from loss of ability to transport them.

Notes: SYNOPSIS. A survey of 41 herbivorous mole-rats, Spalax ehrenbergi Nehring, in Urfa, Adiyaman, and Maras provinces of Turkey revealed 7 new species of Eimeria in addition to previously described Eimeriidae. The shape, average dimensions (in μm) of their oocysts, and the numbers of hosts from which the new species were isolated were as follows: Eimeria urfensis sp. n., ellipsoidal (33 × 21), from 8 hosts; Eimeria adiyamanensis sp. n., ovoid to ellipsoidal (33 × 18), from 6 hosts; Eimeria haranica sp. n., elongate ovoid (37 × 20), from 22 rats; Eimeria marasensis sp. n., ellipsoidal (36 × 18), from 2 rats; Eimeria oytuni sp. n., pear-shaped (24 × 17), from 2 hosts; Eimeria celebii sp. n., ellipsoidal (16 × 9), from 1 rat; and Eimeria torosicum sp. n., spherical to subspherical (11 × 10), from 2 animals.

Notes: Defined media are described that support 14-20 h generation times for Acanthamoeba castellanii and A. rhysodes in monolayer cultures. the media differ in minor ways from previously described media, but the growth rates are greatly improved over previously reported values. Maximum growth rates were observed for A, castellanii in a complex medium containing 21 amino acids, but near-maximum rates could be achieved in relatively simple media containing 9 amino acids. Growth occurred with 6 amino acids, as reported by others, but generation times exceeded 30 h. Amitosis was a common problem during early subcultures in defined media, but became infrequent after repeated transfers. Synchronous encystment resulting in 70-80% cyst formation could be induced in the defined media by glucose and acetate starvation. the rate of encystment varied with cell density at the time of starvation and was optimal at initial densities of 400-800 amebae/mm2.

Notes: Tetrahymena pyriformis strain HSM secretes 4 isozymes of hexosaminidase. Purified isozymes B1 and B2 are eluted from the void volume of a concanavalin A-Sepharose column, suggesting that they are not glycosylated. Purified isozymes A1 and A2 bind to the column and are eluted at ∼0.1 M α-methylmannoside, suggesting that these isozymes are glycoproteins. In agreement with earlier deductions based on a differential kinetic assay for the A and B isozymes, the elution pattern of hexosaminidase activity from material secreted by cells grown to early and late stationary phase was consistent with these secretions containing primarily the B and the A isozymes, respectively.

Notes: The surface proteins and glycoproteins on red cells from normal and Babesia bovis-infected calf blood have been compared. Several radiolabeling probes were used to label specifically external membrane molecules which were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by autoradiography or fluorography. No differences were observed among the Coomassie Blue-stained membrane proteins of erythrocytes from individual uninfected calves. Comparison of red cells from these animals also indicated no qualitative differences in the surface proteins with accessible tyrosyl residues labeled by lactoperoxidase-catalyzed radioiodnation, although some quantitative variation in the uptake of radioactivity into particular proteins was observed. the major radioiodinated bands on normal bovine erythrocytes had Mr of 165, 130, 90, and 45 kiloDaltons. However, labeling of surface glycoproteins by the periodate/[3H]NaBH4 and galactose oxidase (± neuraminidase)/[3H]NaBH4 methods showed significant differences in the surface proteins of red cells from individual uninfected calves. of 14 animals tested, 5 had major labeled glycoproteins of unique Mr. No changes were observed in radioiodinated surface proteins of total red cell samples from infected calves with 0.5-6% parasitemia. Radioiodination of concentrated infected red cells from the same samples (concentrated by selective hypotonic lysis of uninfected erythrocytes in KC1) resulted in the labeling of 3 new surface proteins, with Mr of 118, 115, and 60 kiloDaltons. the same new 125I-labeled bands were identified on infected cells from 3 avirulent strains of B. bovis used in vaccine production. Furthermore, in concentrated infected cells there was very poor radiolabeling of major bands strongly labeled on uninfected cells (Mr 165, 130, and 90 kiloDaltons), suggesting parasite-induced loss of these proteins. Although there were some differences in 3H-labeled surface glycoproteins of red cells from normal and. B. bovis -infected blood, they were restricted to minor labeled bands and were not seen consistently. the labeled surface glycoproteins of concentrated infected cells were very similar to those of the uninfected red blood cells from infected blood.

Notes: SYNOPSIS. Chilomonas paramecium has been studied by light and electron microscopy with special attention to the so-called trichocysts. The electron miscroscope reveals that the anatomy of these bodies is unlike that of the classical trichocysts of ciliates. Since these structures can be extruded from the body of the organism they have been called ejectisomes. The ejectisomes have a regular geometrical shape and a complex internal structure. They consist of two unequal components which are enclosed by a thin membrane. Morphological polarity is established by an anterior smaller unit and a posterior larger unit. When the ejectisomes are found in the peripheral cytoplasm or in a perivestibular position the smaller unit is always oriented toward the surface. A suggestion is made that the ejectisomes are derived from the vesicular component of the Golgi complex. An interpretation is offered concerning a mechanism by which ejectisomes may be extruded from the body.

Notes: SYNOPSIS. Examination of the crop, gizzard, and intestinal contents of chickens fed suspensions of either Eimeria acervulina or E. tenella oocysts and turkeys fed either E. meleagrimitis or E. gallopavonis oocysts indicated that, in all 4 species, (1) oocysts apparently remained unchanged while in the crop, (2) sporocysts were liberated from oocysts while the latter were passing through the gizzard, (3) sporozoites were activated and escaped from liberated sporocysts after they had reached the small intestine, and (4) sporozoites within intact oocysts in the crop, gizzard, and intestines were not activated. In vitro, trypsin 1–300 alone caused a small percentage of sporozoites to excyst from mechanically liberated sporocysts. The percentage of excystation increased greatly when trypsin was added to sodium taurocholate and increased even more when it was combined with chicken or turkey bile.The two duodenal species (E. acervulina and E. meleagrimitis) differed both in vivo and in vitro from the two cecal species (E. tenella and E. gallopavonis). The duodenal species excysted in less time and farther anteriorly in the small intestine than did the cecal species. In addition, sporozoites of the two cecal species survived much longer in media containing trypsin plus bile or sodium taurccholate than did those of the two duodenal species.

Notes: SYNOPSIS. Cysts of the hypotrichous ciliate Pleurotricha lanceolata were placed in dilute aqueous solutions of specific chemicals (carbohydrates, vitamins, L-amino acids, nucleic acids and derivatives, other organic acids and tweens) to assay the effectiveness of these chemicals as excysting agents. It was found that some of the amino acids, deoxyribonucleic acid, ribonucleic acid, and some other organic acids induce excystment at rates equal to or better than the control medium, 0.05% (w/v) aqueous lettuce extract. Preliminary experiments with chelating agents, tri- and tetra-sodium salts of ethylenediaminetetraacetic acid (Versenol 120 and Versene 100), indicate that they successfully initiate excystment, but support it inadequately thereafter so that most of the organisms die within the endocyst.

Notes: SYNOPSIS. A new species of avian malaria parasite is described, for which the name Plasmodium formosanum is proposed. It was observed in the blood of Arboriphila crudigularis, the Formosan hill partridge; other bird species in which it may occur remain unknown, as do the mosquito vectors. It produces very broad, elongate gametocytes, the macrogametocyte usually exhibiting a very large vacuole. The usual number of merozoites is eight. The host cell nucleus typically is not displaced, except laterally by the sexual forms. Only the stages occurring in the blood have yet been seen.

Notes: SYNOPSIS. Three chlorotic Euglena substrains, PBZ-G4, SM-L1, and HB-G, were cultured in light and dark at 25° and 33–35°C. These permanently chlorophyll-less flagellates with different carotenoid distributions were originally derived from a photosynthetic stock culture. Astasia longa, a naturally pigmentless flagellate, and the ciliate Tetrahymena were also employed.None of the organisms were light-dependent at 25°C. At high temperatures only the Euglena substrains showed an effect of light on growth: PBZ-G4 was inhibited, as was the photosynthetic parent strain, whereas SM-L1 and HB-G were stimulated. In all three substrains, monster forms were produced at elevated temperatures; SM-L1 and HB-G were multinucleated, while PBZ-G4 had an enlarged, single nucleus. A small percentage of astasias became multinucleated and almost all the tetrahymenas became giants when cultured at high temperature but neither was affected by light.Light-temperature interactions in the nonphotosynthetic substrains of Euglena are, at present, unexplained. The results are discussed in relation to the carotenoid content and the possible site of interaction. The major factor inducing abnormality is thermal stress.

Notes: SYNOPSIS. Ophryoglena faurei is a large ophryoglena distinguished by a long theront, either cylindrical or pearshaped, by a very bulky, often lengthened trophont and by a voluminous tomont.During fasting, physiological changes cause a secondary and temporary encystment of the theront which lasts for 15–18 days at 18°C. Then the theront becomes globular and cytolyzes. Feeding induces the usual physiological reproductive cycles. Production of 4 and 8 tomites is the result of primary cycles happening before secondary encystment. The secondary cycles induced by feeding of the secondary theront, after its encystment, are characterized by the production of 2 or 4 tomites.If the theront is fed after being starved for about 10 days the resultant trophont encysts and delivers only one theront. The size of this theront is the more reduced as the starvation of the original theront has been longer. The small theronts are able to attain the original major forms but only after several successive alimental cycles and without any secondary encystment of theronts.

Notes: SYNOPSIS. Comparison of dimensional and other morphological details of adult suctorians recovered from the gills of five genera of fish from five widely separated localities in North America indicates that the practice of identifying a distinct suctorian species based upon the species of fish with which it is symbiotic is unfounded. Conversely, identification of suctorian species based on their adult morphology alone under certain conditions is justified. An emended diagnosis of Trichophyra piscium Bütschli, 1889, is presented. The pigment in T. piscium recovered from Oncorhynchus nerka (sockeye salmon) was characterized as melanin and its significance to the suctorians is discussed.