The effects of supraoptimal temperatures on population growth and cortical patterning in Tetrahymena pyriformis and Tetrahymena thermophila: a comparison

In this investigation, we compare the multiplication rates and morphogenetic responses of the two most studied Tetrahymena species, T. pyriformis and T. thermophila, at supraoptimal temperatures. Although the upper temperature limits differ greatly in the two species, the pattern of growth responses to high temperature is for the most part similar, with some differences in detail. The transient recovery of cell division at the highest temperature that allows cell division, characteristic of T. pyriformis, is observed in a less distinct form in T. thermophila. Moreover, there is a remarkable difference in developmental response, with drastic abnormalities in patterning of oral structures during the transient recovery of cell division in T. pyriformis, and far more limited abnormalities under similar conditions in T. thermophila. The abnormalities result from spatial disorder in the alignment and orientation of basal body pairs within the early oral primordium, followed by failures in the realignment that normally occurs as oral structures (membranelles and undulating membrane) mature. Both the initial spatial disorder and the failures in realignment are far more severe in T. pyriformis than in T. thermophila.     

Extrusion of DNA from the macronucleus of Tetrahymena pyriformis GL.

Administration of the thymidine analog 5-bromodeoxyuridine to exponentially growing cultures of Tetrahymena pyriformis GL in chemically defined medium results in inhibition of cell multiplication by at least one generation before DNA synthesis stops. Cell multiplication can be restored in these cultures, if they are transferred to fresh growth medium, but although most of the cells in the culture contain close to a G2-amount of DNA, a full DNA replication round is a prerequisite for renewed cell multiplication. Large extrusion bodies are found at the first division after transfer to fresh growth medium. Autoradiographic analysis has revealed that the DNA in the extrusion body is a representative of the DNA in the macronucleus indicating a random distribution of DNA between daughter nuclei and extrusion body.     

Extrusion of DNA from the Macronucleus of Tetrahymena pyriformis GL

SYNOPSIS Administration of the thymidine analog 5-bromodeoxyuridine to exponentially growing cultures of Tetrahymena pyriformis GL in chemically defined medium results in inhibition of cell multiplication by at least one generation before DNA synthesis stops. Cell multiplication can be restored in these cultures, if they are transferred to fresh growth medium, but although most of the cells in the culture contain close to a G2-amount of DNA, a full DNA replication round is a prerequisite for renewed cell multiplication. Large extrusion bodies are found at the first division after transfer to fresh growth medium. Autoradiographic analysis has revealed that the DNA in the extrusion body is a representative of the DNA in the macronucleus indicating a random distribution of DNA between daughter nuclei and extrusion body.     

Effects of temperature on the growth and survival of the European eel, Anguilla anguilla L.

Experiments to determine the growth rate of eels ( Anguilla anguilla L.) at different temperatures are described and show the optimum temperature for growth to be 22–23° C. The ultimate upper lethal temperature was found to be 38° C and the critical thermal maximum varied from 33 to 39° C for fish acclimated at 14 to 29° C. An attempt was also made to determine lower lethal temperatures. Eels enter a state of torpor at temperatures varying from 3° C for fish acclimated at 29° C to less than 1° C for fish acclimated at 23° C or below. The results have been used to estimate the growth rates expected from eels cultured in power station cooling water using different types of temperature control.     

Synergistic effects of p-fluorophenylalanine and supra-optimal temperatures on cell division in Tetrahymena pyriformis.

The combined effects of sub-optimal (20 degrees) or supra-optimal (31 degrees C and 34 degrees C) temperatures and the amino acid analogue p-fluorophenylalanine on preparations for cell division in Tetrahymena pyriformis were analysed. A supra-optimal temperature of 31 degrees produced a marked exaggeration of the division-delaying capacity of the analogue. The analogue also sensitises cells when combined with the normal heat shock temperature of 34 degrees, which indicates that division proteins continue to be synthesized at this temperature but cannot be effectively utilized by the cell. These results support the model that preparations involve interactions between and formation of assemblies of proteins on a strict time schedule shortly before division.     

Comparison of the immunochemical responses by anti-NADPH-cytochrome c reductase of Tetrahymena pyriformis (strain NT-1) in protozoan cells and mammalian liver microsomes

Comparison of the microsomal NADPH-cytochrome c reductase activities in the four Tetrahymena cells (pyriformis, strain GL and NT-1; thermophilia; ISO) and rat liver was studied. The reductase activity in strain NT-1 was lowest among four Tetrahymena cells grown at 24 degrees C. Rabbit antibody was prepared against the purified NADPH-cytochrome c reductase from Tetrahymena pyriformis (strain NT-1) microsomes. Microsomal NADPH-cytochrome c reductase activities in various Tetrahymena cells were inhibited in proportion to the amount of antibody added, in the order of GL greater than NT-1 greater than thermophilia greater than ISO. No inhibition of reductase activity by antibody was observed in rat liver microsomes.     

The Effects of Temperature and Heavy Water on Cell Division in Heat-Synchronized Cells of Tetrahymena*

SYNOPSIS. A study was undertaken of the rates of cell division of heat-synchronized cells of Tetrahymena pyriformis GL at various temperatures in water and 20, 30 and 40% heavy water. The results suggest that division rate is limited by a protein which undergoes both high and low temperature denaturation and that this protein is partially stabilized against heat-denaturation while becoming more susceptible to cold denaturation in the presence of heavy water. Thus, the optimum temperature for division shifts upward as the heavy water concentration is increased, with a maximum shift of 1 C occurring in 40% heavy water. In addition, division activity occurs in heavy water at 34 C, a temperature at which cells kept in water are blocked. Furthermore, the sharp increase in slope seen in the low temperature portion of Arrhenius plots of the data, occurs at higher temperatures when heavy water is present. Finally, at virtually all temperatures, except the highest, heavy water has a depressive effect on division rate indicating a general inhibitory influence of deuterium-substituted water on rate processes within the cell.     

A mathematical model of the growth of a population of Legionella pneumophila in the presence of Tetrahymena pyriformis protozoa]

The mathematical model describing the dynamics of the growth of L. pneumophila in aqueous environment in the presence of protozoa has been worked out. The model has demonstrated considerable heterogeneity of the initial population of virulent L. pneumophila strains. The number of bacteria capable of multiplication in Infusoria is no more than 0.1% of the initial population. The time of the generation of the infective agent inside Tetrahymena pyriformis is 2.8 hours.     

Growth, respiration and survival of Legionella pneumophila at high temperatures

The optimum temperature for multiplication of legionella strains in culture media is around 37°C. The effect of high temperatures on the growth of strains isolated from various environments is poorly known. We studied the growth (cell multiplication, respiration) of clinical and environmental Legionella pneumophila strains in liquid media at intervals of 0.5°C in the temperature range from 41.6 to 51.6°C using a temperature gradient incubator. Cell multiplication and CO₂ production decreased markedly with all the strains at temperatures above 44–45°C. CO₂ continued to be produced up to 51.6C even if cell multiplication generally stopped at around 48.4–50.0C. Thus, legionella retained its metabolic activity beyond the maximum temperature for cell multiplication. The CO₂ production per bacterial cell (metabolic quotient, qCO₂) increased with increasing temperature up to 45°C, whereafter it decreased, the turning point being almost at the same at which the rate of cell multiplication decreased. The difference in qCO₂ between the strains may reflect their different physiological capacities for tolerating high temperatures.     

The study of Salmonellaenteritidis growth kinetics using Rapid Automated Bacterial Impedance Technique

In order to study strain-specific differences in their growth behaviour at different, and particularly lower, temperatures, generation times for 45 strains of Salmonella enteritidis isolated from food were determined impedimetrically over a temperature range from 7 to 42 °C. In practical terms, 7 °C is the minimum requirement for Salm. enteritidis growth, and generation time variability increases markedly as this temperature is reached. Reports in the literature describing psychrotrophic behaviour and multiplication at lower temperatures cannot be confirmed. Generation time variability increased as temperature moved away from the optimal range with variation coefficients tending to rise as temperature fell. The great variability of multiplication parameters near the growth limit found in Salm. enteritidis may also be a characteristic of other bacterial species. It is therefore imperative to commence studies on larger numbers of strains to allow prediction of their behaviour at lower temperatures.     

Replicon size and rate of DNA replication in the macronucleus of Tetrahymena pyriformis]

The size of replication units (or replicons) measured in Tetrahymena pyriformis GL macronuclear DNA reaches 20--30 microns, according to the two independent methods: DNA fiber autoradiography, and alkaline isokinetic sucrose gradient centrifugation. The synthesis of new DNA fragments--replicons and their subsequent assembly are separated by time intervals (30 min). The rate of DNA synthesis for one fork averaged 0.6--0.7 microns/min. These data were obtained for cells of cultures being both in the expotential phase of growth, and those synchronized by starvation-refeeding. The generation time of T. pyriformis cells, calculated by the increase of the part of labeled nuclei, is almost 2 hours; the synthesis lasts 1 hour. Total amount of replication units in polyploid (polygenomic) Tetrahymena macronucleus is about 3000. Their initiation during S-period is presumably asynchronous.     

Biological parameters of Thripobius semiluteus Bouček (Hym., Eulophidae), a larval endoparasitoid of Heliothrips haemorrhoidalis (Bouché) (Thysan., Thripidae)

Abstract:  The endoparasitoid Thripobius semiluteus Bouček was recently introduced to Italy from Israel for the biological control of Heliothrips haemorrhoidalis (Bouché). In this study, some aspects of the biology of T. semiluteus were determined in the laboratory. Developmental time (egg to adult), potential fecundity, realized fecundity, progeny, daily rate of deposition of eggs and several demographic growth parameters were evaluated. Studies of the longevity of fed and starved adults at seven temperatures (3, 10, 15, 20, 23, 25 and 30 ± 1°C) showed that fed T. semiluteus lived longest at 15°C (36.45 days) and shortest at 3°C (1.45 days). Longevity was reduced significantly at higher and lower temperatures than 15°C and when honey was not provided. Development time was measured at six temperatures. Pupae did not complete development at 10°C. The lower theoretical temperature threshold was 9.0°C; the optimum developmental temperature and the upper lethal threshold were 28.0°C and 34.1°C respectively. The most rapid development was found at 30°C, but waSPS suffered significantly higher mortality at this temperature than at other temperatures. Ovigeny index showed that T. semiluteus is synovigenic, with a mean realized fecundity of 78.8 eggs per female. The mean progeny was 68.2 adults per female on fed adults.     

Monstrous Tetrahymena with intraclonal variation in structure produced by hereditary modification of normal cells.

Monstrous Tetrahymena pyriformis strain GL may be isolated after exposure of normal cells to numerous heat shocks, to flattening on agar or gelatin plates, or to viscous solutions of methyl cellouse. It is shown that in some cases the abnormalities are inherited and that this results in clones where the cells are different from each other and have various abnormalities with respect to cortical pattern, swimming and feeding behavior, and generation time. Furthermore, it is shown that these cells are produced rather than selected by the experimental treatments. Evidence is presented that growth without division is important for production of the abnormal organisms. The basis of the inheritance of the abnormalities is discussed.     

Monstrous Tetrahymena with Intraclonal Variation in Structure Produced by Hereditary Modification of Normal Cells

SYNOPSIS Monstrous Tetrahymena pyriformis strain GL may be isolated after exposure of normal cells to numerous heat shocks, to flattening on agar or gelatin plates, or to viscous solutions of methyl cellulose. It is shown that in some cases the abnormalities are inherited and that this results in clones where the cells are different from each other and have various abnormalities with respect to cortical pattern, swimming and feeding behavior, and generation time. Furthermore, it is shown that these cells are produced rather than selected by the experimental treatments. Evidence is presented that growth without division is important for production of the abnormal organisms. The basis of the inheritance of the abnormalities is discussed.     

The critical thermal limits for juvenile Arctic charr Salvelinus alpinus

The chief objective was to determine the critical thermal limits for alevins, fry and parr of Arctic charr, Salvelinus alpinus , (L.) from four races living in Windermere (northwest England). The experimental fish were reared in a hatchery but were the progeny of wild parents. As comparisons between tethal temperatures at four acclimation temperatures (5, 10, 15, 20° C) revealed few significant racial differences, the data were pooled to estimate the lethal values for survival over 7 days (incipient lethal temperature) and over only 10 min (ultimate lethal temperature) for each life stage. Upper lethal values increased with acclimation temperatures for alevins but this effect was negligible for fry and parr, Alevins were generally less tolerant than fry and parr at lower, but not higher, acclimation temperatures; e.g. after acclimation at 5° C, mean upper ultimate values were 23·3, 25·1 and 25·7° C and mean upper incipient values were 18·7, 21·5 and 21·5° C for alevins, fry and parr respectively; after acclimation at 20° C, mean upper ultimate and incipient values were 26·2, 26·1 and 26·6° C and 20·8, 20·8 and 21·6° C for alevins, fry and parr respectively. The area of the temperature tolerance polygon (expressed as ° C²) for juvenile Arctic charr is amongst the lowest recorded for salmonids; being 409, 439 and 461° C² for alevins, fry and parr respectively. These low values are due to lower upper tolerance limits, not high lower tolerance limits; the latter being close to 0° C (<1°C for parr and fry, <0·3° C for alevins) at all acclimation temperatures. Arctic charr are therefore amongst the least resistant of salmonids to high temperatures but probably the most resistant to low temperatures.     

Thermal resistance and upper lethal temperatures of underyearling Lake Inari Arctic charr

Underyearling Arctic charr were acclimated to six temperatures between 6 and 21·5°C and thermal tolerance and resistance were tested after an acclimation period of at least 2 weeks. Resistance times were influenced by acclimation temperature and the highest upper incipient lethal temperature was 23–24°C. An upper limit for cultivation of Lake Inari charr is suggested to be 21°C which is the intercept of the function which represents the upper limit of the thermal tolerance zone.     

Induction of division synchrony in Tetrahymena pyriformis by a single hypoxic shock. Its use in elucidating control of the cell cycle by adenosine 3':5'-monophosphate.

A single hypoxic shock was used to induce division synchrony in Tetrahymena pyriformis. Hypoxia results in accumulation of the cells in the G2 phase of the cell cycle. Cyclic AMP and adenyl cyclase activity were measured in this system. Cell-cycle blockade was associated with extraordinarily high levels of intracellular cyclic AMP. After release from hypoxia, the cells retain a characteristic pattern of modulation of cyclic AMP associated with division that is found in selection-synchronized cells. The results are discussed with reference to other methods of induction synchrony and related studies on the natural cell cycle in this organism.     

Characteristics of alternating temperatures which stimulate loss of dormancy in seeds of Rumex obtusifolius L. and Rumex crispus L.

Abstract. Alternating temperatures stimulate the germination of Rumex crispus L. and Rumex obtusifolius L. The optimum period spent at the lower temperature in a diurnal cycle is greater than that spent at the higher temperature. Under most conditions the optimum period at the upper temperatures is about 8 h but, as the upper temperature of a cycle is increased, the optimum period at the upper temperature becomes shorter and more critical. Thus when it is 35°C the optimum period is 2.5–4 h in the light, or about 1 h in the dark. The effect of alternating temperatures is much less in the dark than in the light and in general only extreme alternations with short periods at the higher temperature are effective in the dark. In the light any temperature alternation within the range 1–35°C is effective to at least some extent, providing the temperature difference is 5°C or more and providing the alternation includes one temperature which is above approximately 15°C and one which is below approximately 25°C. The optimum temperature difference is about 15°C. In the light, 4 to 10 cycles saturate the response, but in the dark, where the effect is much less, the response may not be saturated even by 16 cycles. KNO₃ at 10⁻³ M has little effect on the response to alternating temperatures either in the light or the dark. The response to alternating temperature regimes does not appear to vary in quality, i.e., in terms of which particular treatments are best, but it varies in magnitude with site and year of seed collection; and it increases slowly during dry storage, even when stored at a temperature as low as 1.5°C.     

Temperature Effects on Maturity Periods in Tetrahymena pyriformis Syngen 1*

Cells of Tetrahymena pyriformis syngen 1 grown at 30 C after conjugation achieve sexual maturity more quickly than do cells grown at 19 C, whether time is measured in numbers of cell divisions or in terms of absolute time. This result is achieved regardless of the temperature at which conjugation and nuclear reorganization occur. These observations differ from those of other workers investigating Paramecium, and suggest that the long term “chronometer” is more tightly coupled to cell division in Paramecium multimicronucleatum and Paramecium caudatum than in Tetrahymena pyriformis.