Cytotoxicity of synthetic fuel products on Tetrahymena pyriformis. I. Phenol.

Phenol is the major organic constituent of coal-conversion scrub water and is therfore a potential environmental contaminant. After Tetrahymena pyriformis strain GL-C, syngen 1 was exposed to phenol, its behavior, cytology, respiration, and growth rates were examined. Concentrations larger than or equal to 75 mg/liter alter cell motility, shape, and contractile vacuole activity. O2 uptake was abruptly reduced within 3 min of exposure to phenol in concentrations as low as 10 mg/liter. Concomitantly there was an increase in the electron density of the mitochondrial matrix. Recovery to normal rates of O2 consumption was paralleled by a return to normal matrix density. Alterations of mucocysts, pellicle, and glycogen were also observed. The length of lag phase growth curves increased generally in proportion to concentration of toxicant. Phenol, however, did not affect the rate of cell multiplication during the exponential growth phase. The potential use of this system to examine the effects of other possible organic pollutants is discussed.     

Cytotoxicity of Synthetic Fuel Products on Tetrahymena pyriformis. I. Phenol*

SYNOPSIS. Phenol is the major organic constituent of coal-conversion scrub water and is therefore a potential environmental contaminant. After Tetrahymena pyriformis strain GL-C, syngen 1 was exposed to phenol, its behavior, cytology, respiration, and growth rates were examined. Concentrations ≥ 75 mg/liter alter cell motility, shape, and contractile vacuole activity. O₂ uptake was abruptly reduced within 3 min of exposure to phenol in concentrations as low as 10 mg/liter. Concomitantly there was an increase in the electron density of the mitochondrial matrix. Recovery to normal rates of O₂ consumption was paralleled by a return to normal matrix density. Alterations of mucocysts, pellicle, and glycogen were also observed. The length of lag phase growth curves increased generally in proportion to concentration of toxicant. Phenol, however, did not affect the rate of cell multiplication during the exponential growth phase. The potential use of this system to examine the effects of other possible organic pollutants is discussed.     

Proliferation of Legionella pneumophila as an intracellular parasite of the ciliated protozoan Tetrahymena pyriformis

In a series of experiments, we have determined that Legionella pneumophila will proliferate as an intracellular parasite of the ciliated holotrich Tetrahymena pyriformis in sterile tap water at 35 degrees C. After 7 days of incubation, serpentine chains of approximately 10(3) L. pneumophila cells were observed throughout the cytoplasm of the protozoan infected initially with 1 to 30 L. pneumophila cells. The overall L. pneumophila population increased from ca. 1.0 X 10(2) to ca. 5.0 X 10(4) cells per ml in the coculture within this time frame. The interactions between the protozoan and the bacterium appear to depend upon their concentrations as well as temperature of incubation. L. pneumophila did not multiply in sterile tap water alone, in suspensions of lysed T. pyriformis, or in cell-free filtrates of a T. pyriformis culture. In addition to establishing an ecological model, we found that addition of T. pyriformis to environmental specimens served as an enrichment method that improved isolation of legionella from the specimens.     

Proliferation of Legionella pneumophila as an intracellular parasite of the ciliated protozoan Tetrahymena pyriformis.

In a series of experiments, we have determined that Legionella pneumophila will proliferate as an intracellular parasite of the ciliated holotrich Tetrahymena pyriformis in sterile tap water at 35 degrees C. After 7 days of incubation, serpentine chains of approximately 10(3) L. pneumophila cells were observed throughout the cytoplasm of the protozoan infected initially with 1 to 30 L. pneumophila cells. The overall L. pneumophila population increased from ca. 1.0 X 10(2) to ca. 5.0 X 10(4) cells per ml in the coculture within this time frame. The interactions between the protozoan and the bacterium appear to depend upon their concentrations as well as temperature of incubation. L. pneumophila did not multiply in sterile tap water alone, in suspensions of lysed T. pyriformis, or in cell-free filtrates of a T. pyriformis culture. In addition to establishing an ecological model, we found that addition of T. pyriformis to environmental specimens served as an enrichment method that improved isolation of legionella from the specimens.     

Isocitrate dehydrogenase of Tetrahymena pyriformis

Summary We have studied the isocitrate dehydrogenase ofTetrahymena pyriformis. This enzyme is able to utilize both NAD and NADP, but kinetic studies suggest that the enzymatic activity with NAD is not of physiological significance.Some of the factors that might regulate the NADP-dependent isocitrate dehydrogenase were also studied. This enzyme has an absolute requirement for divalent cations; Mg²⁺ and Mn²⁺ will serve as cofactors but the latter is more effective than the former.It is known that this enzyme is subject to a concerted inhibition by oxaloacetate and glyoxylate. Either glyoxylate or oxaloacetate alone also are capable of inhibiting the enzyme although higher concentrations are required. We have found concerted inhibition also for the NAD-dependent isocitrate dehydrogenase from rat liver and yeast. The activity of theTetrahymena pyriformis enzyme is inhibited by NADPH. This inhibition is competitive with NADP. The Ki and Km values are, respectively, 23µ m and 18µ m.     

Cytotoxicity of synthetic fuel products on Tetrahymena pyriformis. II. Shale oil retort water.

Shale oil retort water is obtained by centrifuging the oil/water emulsion produced by oil shale retorting. The ciliate Tetrahymena pyriformis was exposed to retort water; 2, 1, and 0.5% initially increased motility; longer exposures decreased motility. Three, 4, and 5% all decreased motility. Cell lysis was directly related to concentration; after 24 h, population densities were 0, 10, and 25% of controls for 2, 1, and 0.5% retort water, respectively. Oxygen consumption paralleled the motility pattern: at lower concentrations it increased initially but decreased with extended exposures while at higher concentrations it decreased rapidly. The most striking cytologic alteration of cells exposed to the toxicant occurred in the membranes; alterations of mucocysts and glycogen content were also observed, but mitochondrial changes were not. Population growth was affected at much lower concentrations than the other test indices. The growth of test populations reached a plateau at values inversely related to concentration: concentrations less than 0.4% had no effect on growth rate.     

Characteristics of the interaction of legionellae and Tetrahymena pyriformis

L. pneumophila avirulent strains have been shown to lose their capacity for multiplication in T. pyriformis, while the concentration of the virulent strain increases 1000-fold. The loss of the hemolytic activity of L. pneumophila virulent strain leads to the loss of its capacity for multiplication in infusorians.     

Intracellular phospholipase activities of Tetrahymena pyriformis

Phospholipase activity was studied in the protozoan Tetrahymena pyriformis NT-1 by using exogenous phosphatidylethanolamine and phosphatidylcholine. Several phospholipase activities were found in Tetrahymena homogenates. They were distinguished with respect to pH optimum, activity dependence on Ca2+, substrate specificity and positional specificity. Ca2+-Dependent phospholipase activity had an optimal pH around 9 and gave rise to free fatty acid and lysophospholipid. This enzyme hydrolyzes phosphatidylethanolamine but not phosphatidylcholine. The alkaline phospholipase with A1 activity was located mainly in the surface membrane (pellicle fraction). The enzyme activity had a pH optimum ranging from 8 to 9, and required 2 mM CaCl2 for the maximal activity. All detergents tested inhibited the enzyme activity. Ca2+-Independent phospholipase activity had an optimal pH from 4 to 5 and gave rise to free fatty acid, lysophospholipid, diacylglycerol, and monoacylglycerol. We concluded that there are at least three phospholipase in Tetrahymena homogenates, i.e., alkaline phospholipase A and acidic phospholipases A and C.     

The Distribution of Tetrahymena pyriformis1

SYNOPSIS. This paper is a brief account of both amicronucleate and sexually active strains of Tetrahymena pyriformis and their distribution with some comments on their possible evolution.     

Serotype Expression and Transformation in Tetrahymena pyriformis

SYNOPSIS. Of the 4 serotypes of Tetrahymena pyriformis, syngen 1, the H (expressed at 20–35 C) and T (expressed at 36–40 C) serotypes and their genetic bases have received the most attention. The present study describes the L and I serotypes, considers their transformations to and from the H serotype, and compares inter-locus repression with allelic repression. The L serotype occurs below 20 C. No strain differences have been found between L antigens. However, the rate of transformation between H and L varies in the families, which can be ranked according to their preference for the H or L serotype. A3 is strongly L; D1 is strongly H; the other families form a continuum between these extremes. Preference for the H or L serotype is not correlated with the H allele present. Serotype differences may be hereditary, by the “dilution” test for hereditary differences. Altho the environment largely determines which serotype will be expressed, sublines can maintain established H and L serotypes for more than 40 fissions in a common environment. Crosses between H and L cells suggest macronuclear control of serotype determination and a cytoplasmic influence on macronuclear differentiation. Evidence for the cytoplasmic influence comes from both conjugants and nonconjugants. The I serotype is induced from the H serotype at room temperature by treatment with specific H antibodies, which are also necessary for I maintenance. The I serotype consists of a series of unstable serotypes which transform among themselves. Clones go thru an initial I type one day after induction and then diversify to express secondary types. The initial type and the secondary types are characteristic of the family but are not related to the H genotype. Family C1 lacks 2 secondary types found in other families studied. The differences between inter-locus repression and allelic repression are marked. Altho inter-locus repression is labile, all efforts to reverse allelic differentiations have failed. H heterozygotes maintained for long periods of time in the I or L state showed no reversal of H allelic repression.     

Phospholipase D Activity in the Tetrahymena pyriformis GL

Phospholipase D (PLD) is an enzyme which participates in the signalling mechanism cleaving phosphatidylcholine (PC) to choline and phosphatidic acid (PA). In Tetrahymena pyriformis GL this enzyme activity is enhanced by different kinds of agonists (sodium orthovanadate, sodium fluoride and phorbol 12-myristate 13-acetate), and its activity can be inhibited by inhibitors such as pertussis toxin, calphostin C, genistein, trifluoperazine. These results suggest that the PLD signalling pathway is connected with the tyrosine kinase, phospholipase C, phosphatidylinositol and G-protein coupled signalling pathways. By demonstrating the PLD activity in Tetrahymena our knowledge on the signalling mechanisms at a unicellular level has been extended. The results support our view that most transducing mechanisms that are characteristic of mammalian cells are also in the protozoan Tetrahymena. © 1997 John Wiley & Sons, Ltd.