Effect of Acetate on Esterase C Activity During the Growth Cycle of Paramecium*

SYNOPSIS Enhanced esterase C activity could be demonstrated by starch gel electrophoresis in various stocks of Paramecium spp. (P. primaurelia stocks 90 and 540, P. biaurelia stock 93, P. tetraurelia stock 29. P. pentaurelia stock 87, P. octaurelia stocks 31 and 300, and P. multimicronucleatum species 3, stock 8 MO) grown in Adaptation Medium. This esterase, however, was barely detectable when they were cultivated in Axenic Medium. Addition of trypticase to Adaptation Medium resulted in reduction of esterase C in the ciliates. This effect is ascribable to Na acetate present in trypticase. Since esterase C increased with the decrease in acetate concentration (as estimated by gas-liquid chromatography) during growth of Paramecium, acetate appears to be utilized by the cells. Sensitivity of esterase C to acetate occurs in all 6 species of Paramecium examined. Different stocks within a species may have different levels of sensitivity; in one case this is genetically determined. The results emphasize the importance of controlling and manipulating growth conditions for the assessment of inter- and intraspecies variations in the isozymes of Paramecium.     

Interspecies Relationships in the Paramecium aurelia Complex: Acid Phosphatase Variation1

ABSTRACT. Up to five zones of acid phosphatase activity appear in gels after electrophoresis of detergent-treated extracts from 13 of the 14 species of the Paramecium aurelia complex. The overall pattern is somewhat similar for all species: differences in intensity and mobility of individual zones permit the grouping of these sibling species into eight groups. All 14 species can be identified using the procedure of enzyme electrophoresis, although two of them are more similar than is usually the case. Problems of misclassification are discussed in terms of the nature and frequency of variants. With the judicious choice of enzymes used to screen new stocks, these problems can be circumvented. Species relationships are updated using 11 enzymes. A dendrogram constructed from the matrix of genetic distances shows four clusters of species: (i) P. biaurelia, P. triaurelia; (ii) P. primaurelia, P. pentaurelia, P. sexaurelia, P. novaurelia; (iii) P. septaurelia, P. undecaurelia, P. tredecaurelia, P. quadecaurelia; and (iv) P. tetraurelia, P. octaurelia, P. decaurelia, P. dodecaurelia. Distances between the species are large, on the order of the differences between Drosophila species. The species are characterized by an extraordinary lack of geographical differentiation and great morphological similarity, which contrasts strongly with the molecular differentiation.     

Leaf‐litter breakdown in pasture and deciduous woodland streams: a comparison among three European regions

1. Human land‐use has altered catchments on a large scale in most parts of the world, with one of the most profound changes relevant for streams and rivers being the widespread clearance of woody riparian vegetation to make way for livestock grazing pasture. Increasingly, environmental legislation, such as the EU Water Framework Directive (EU WFD), calls for bioassessment tools that can detect such anthropogenic impacts on ecosystem functioning. 2. We conducted a large‐scale field experiment in 30 European streams to quantify leaf‐litter breakdown, a key ecosystem process, in streams whose riparian zones and catchments had been cleared for pasture compared with those in native deciduous woodland. The study encompassed a west–east gradient, from Ireland to Switzerland to Romania, with each of the three countries representing a distinct region. We used coarse‐mesh and fine‐mesh litter bags (10 and 0.5 mm, respectively) to assess total, microbial and, by difference, macroinvertebrate‐mediated breakdown. 3. Overall, total breakdown rates did not differ between land‐use categories, but in some regions macroinvertebrate‐mediated breakdown was higher in deciduous woodland streams, whereas microbial breakdown was higher in pasture streams. This result suggests that overall ecosystem functioning is maintained by compensatory increases in microbial activity in pasture streams. 4. We suggest that simple coefficients of breakdown rates on their own often might not be powerful enough as a bioassessment tool for detecting differences related to land‐use such as riparian vegetation removal. However, shifts in the relative contributions to breakdown by microbial decomposers versus invertebrate detritivores, as revealed by the ratios of their associated breakdown rate coefficients, showed clear responses to land‐use.     

Climate‐driven increases in storm frequency simplify kelp forest food webs

Climate models predict a dramatic increase in the annual frequency and severity of extreme weather events during the next century. Here we show that increases in the annual frequency of severe storms lead to a decrease in the diversity and complexity of food webs of giant kelp forests, one of the most productive habitats on Earth. We demonstrate this by linking natural variation in storms with measured changes in kelp forest food web structure in the Santa Barbara Channel using structural equation modeling (SEM). We then match predictions from statistical models to results from a multiyear kelp removal experiment designed to simulate frequent large storms. Both SEM models and experiments agree: if large storms remain at their current annual frequency (roughly one major kelp‐removing storm every 3.5 years), periodic storms help maintain the complexity of kelp forest food webs. However, if large storms increase in annual frequency and begin to occur year after year, kelp forest food webs become less diverse and complex as species go locally extinct. The loss of complexity occurs primarily due to decreases in the diversity and complexity of higher trophic levels. Our findings demonstrate that shifts in climate‐driven disturbances that affect foundation species are likely to have impacts that cascade through entire ecosystems.     

Macro-zooplankter responses to simulated climate warming in experimental freshwater microcosms

1. We report data collected from 48 replicated microcosm communities created to mimic plant‐dominated shallow lake and pond environments. Over a 2‐year period, the microcosms were subjected to warming treatments (continuous 3 °C above ambient and 3 °C above ambient during summer only), a nutrient addition treatment and the presence or absence of fish. We tracked macro‐zooplankter dynamics, censusing cladoceran populations at the species level, copepods at the order level and ostracods as a class. 2. Responses to warming were subtle. Cladoceran diversity and overall abundance were not significantly affected by warming, although measures of community evenness increased. Warming effects on patterns of population trajectories tended to be strongly seasonal and most apparent during periods of pronounced increase. Populations of the prevalent cladocerans, Chydorus sphaericus and Simocephalus vetulus, displayed idiosyncratic patterns, with evidence in the case of S. vetulus for a negative relationship between warming and body‐size at maturity. Copepod populations were reduced in size by warming, but those of ostracods increased. 3. The effects of the nutrient addition and fish treatments were strong and consistent, interacting little with warming effects in statistical models. Zooplankter abundance tended to be the highest in the fish‐free microcosms receiving additional nutrient inputs and lowest when fish were present and no nutrients were added. Both treatments reduced cladoceran diversity and community evenness. 4. We suggest that warming, independently, is unlikely to supplant the effects of changing nutrient loading and fish predation as the major driver of zooplankter dynamics in shallow lakes and ponds. Moreover, in the situations where warming was of significant influence in our experiment, the distinction between summer‐only warming and year‐around warming was blurred. This suggests that warming effects were most pervasive during the summer, at the upper end of the temperature spectrum.     

The Purification of DNA from the Genomes of Paramecium aurelia and Tetrahymena pyriformis1

SYNOPSIS. Methods are described for the recovery of highly purified DNA from Paramecium aurelia and Tetrahymena pyriformis in high yields. Our DNA is only slightly contaminated with RNA and carbohydrate, and little or no protein can be detected. We could not reduce the RNA (orcinol-positive material) by further treatment (sephadex or hydroxyapatite chromatography and preparative CsCl gradients). At the extreme our DNA is contaminated with 15–20% RNA but the real value is most likely considerably lower than this. The DNA we have prepared from Paramecium and Tetrahymena shows all the properties of double-stranded, high molecular weight DNA when characterized by temperature melting, CsCl density gradient centrifugation and hydroxyapatite and sephadex chromatography. When denatured, it absorbs to nitrocellulose filters. The 2 major results of importance from our work reported here are: (1) There is similarity in base composition of DNA from different syngens of Paramecium (28% G+C for syngens 1, 2, 4, 5, and 9 and 29–30% G+C for syngen 8) while there is variation between the syngens of Tetrahymena (24–31% G+C for syngens 1, 4, 7, 10, 11, and 12); (2) the density of any Paramecium DNA varies depending upon whether the cells are grown in the presence of bacteria or in axenic medium. Our results are compatible with observations previously reported for Tetrahymena but contradict those made for Paramecium. The earlier reports of differences in base composition between syngens of Paramecium are probably due in large part to the use of stocks grown on bacteria.