Stock cultures of free-living amebas: effect of temperature on viability and pathogenicity

The effect of temperature on the viability and pathogenicity of free-living amebas during long-term storage were carried out for 34 mo at room temperature, 40 mo at +4 C, and 15 mo at -15 C. During long-term storage, pathogenic amebas tend to lose their virulence. Acanthamoeba showed better survival potential than Naegleria.     

Comparison of the response of in hospite and ex hospite Symbiodinium to elevated temperature

Corals exhibit different responses to increasing temperature but it remains unclear whether this is determined exclusively by symbiont type or by intrinsic properties of the host. Here, we investigated the response to elevated temperature of symbionts of the same ITS2 type from three acroporid species from Bolinao, northwestern Philippines. Maximum quantum yield (F_v/F_m) of PSII was measured in symbionts subjected to 26°C (average winter temperature) and 31°C (average summer temperature) for up to 48 hrs. Greater reduction in F_v/F_m was observed for ex hospite than for in hospite symbionts. However, no significant differences in response could be discerned for symbionts associated with or originating from the different acroporids. Thus, while these findings confirm that the coral host can protect in hospite symbionts from temperature perturbations, for the acroporids in this study, there is still no evidence that host type confers differential thermal susceptibility to the symbionts or to the coral holobiont.     

The effect of growth temperature on the bioenergetics of photosynthetic algal cultures

Two photosynthetic algal cultures, one Chlorella vulgaris, and the other a Chlorogonium sp., were cultured under light limitations in chemostats. The effects of growth temperature on their energy yield and maintenance energy requirement were studied. It was observed that a lowering in temperature resulted in a lower maximum growth yield from the light energy, Y(G). This was attributed to two reasons. First, at low temperatures there was a change in the algal cell composition with more energy being expended to synthesize a higher biomass protein content. Secondly, at low temperatures, a cyanide-resistant respiratory pathway became operative which led to a decrease in the number of ATP being generated. The maintenance energy coefficient was a function of temperature increasing with decreasing temperature. This might reflect energy wastage by the cell at low temperatures. The maximum specific growth rate dropped with decreasing temperature, and can be described by an Arrhenius type rate-temperature model up to the optimal temperature for growth; i.e., activation energy remained constant.     

Genetic diversity of free-living Symbiodinium in surface water and sediment of Hawai‘i and Florida

Marine dinoflagellates in the genus Symbiodinium are primarily known for their symbiotic associations with invertebrates and protists, although they are also found free-living in nanoplankton and microphytobenthic communities. Free-living Symbiodinium are necessary for hosts that must acquire their symbionts anew each generation and for the possible reestablishment of endosymbiosis in bleached adults. The diversity and ecology of free-living Symbiodinium are not well studied by comparison with their endosymbiotic counterparts, and as a result, our understanding of the linkages between free-living and endosymbiotic Symbiodinium is poor. Here, we begin to address this knowledge gap by describing the genetic diversity of Symbiodinium in the surface water and reef sediments of Hawai‘i and Florida using Symbiodinium-specific primers for the hypervariable region of the chloroplast 23S domain V (cp23S-HVR). In total, 29 Symbiodinium sequence types were detected, 16 of which were novel. The majority of Symbiodinium sequence types in free-living environments belonged to clades A and B, but smaller numbers of sequence types belonging to clades C, D, and G were also detected. The majority of sequences recovered from Hawai‘i belonged to clades A and C and those from Florida to clade B. Such distribution patterns are consistent with the endosymbiotic diversity previously reported for these two regions. The ancestral sequence types in each clade were typically recovered from surface water and sediments both in Hawai‘i and Florida and have been previously reported as endosymbionts of a range of invertebrates, suggesting that these types have the capacity to exploit a range of very different habitats. More derived sequence types in clades A, B, C, and G were not recovered here, suggesting they are potentially restricted to endosymbiotic environments.     

Assessment of metabolic modulation in free-living versus endosymbiotic Symbiodinium using synchrotron radiation-based infrared microspectroscopy

The endosymbiotic relationship between coral hosts and dinoflagellates of the genus Symbiodinium is critical for the growth and productivity of coral reef ecosystems. Here, synchrotron radiation-based infrared microspectroscopy was applied to examine metabolite concentration differences between endosymbiotic (within the anemone Aiptasia pulchella) and free-living Symbiodinium over the light-dark cycle. Significant differences in levels of lipids, nitrogenous compounds, polysaccharides and putative cell wall components were documented. Compared with free-living Symbiodinium, total lipids, unsaturated lipids and polysaccharides were relatively enriched in endosymbiotic Symbiodinium during both light and dark photoperiods. Concentrations of cell wall-related metabolites did not vary temporally in endosymbiotic samples; in contrast, the concentrations of these metabolites increased dramatically during the dark photoperiod in free-living samples, possibly reflecting rhythmic cell-wall synthesis related to light-driven cell proliferation. The level of nitrogenous compounds in endosymbiotic cells did not vary greatly across the light-dark cycle and in general was significantly lower than that observed in free-living samples collected during the light. Collectively, these data suggest that nitrogen limitation is a factor that the host cell exploits to induce the biosynthesis of lipids and polysaccharides in endosymbiotic Symbiodinium.     

The effect of elevated temperature on the complexation of am3+ with chloride

The temperature dependence of the formation of the 1:1 Am³⁺-Cl^- solution complex is reported. Stability constants were determined as a function of temperature using a solvent extraction technique. The stability constant for the formation of the AmCl²⁺ complex is measurably higher at 50‡C compared to values determined under ambient conditions. From these studies, we have estimated the enthalpy of complexation in this temperature range. A small, positive enthalpy was observed. The results are discussed in the context of previous studies.     

The effect of elevated temperature on the aggregation of African green monkey kidney cells

The aggregation kinetics of African green monkey kidney cells CV1 and of the SV40 transformed derivative COS1 cells that had been incubated at 37 degrees C or 43.5 degrees C was studied using the shaking flask system. COS1 cells show a three fold decrease in aggregation rate compared to CV1 cells when both cell types were incubated and aggregated at 37 degrees C. When these cell types were incubated at 43.5 degrees C for 5 hours, then aggregated at 37 degrees C showed a faster aggregation kinetics than before. Their aggregation at 43.5 degrees C with prior incubation at 37 degrees C or 43.5 degrees C reached the aggregation kinetics of 43.5 degrees C incubated cells aggregated at 37 degrees C. The addition of serum in the aggregation medium did not influence extensively the aggregation rates of both cell types.     

The effect of elevated temperature on spawning of introduced pumpkinseed Lepomis gibbosus in Europe

In an experimental comparison of pumpkinseed Lepomis gibbosus reproduction under ambient and climate change water temperature regimes, spawning occurred earlier in the season, which is likely to lead to greater young-of-the-year survival with concomitant implications in the U.K. under warmer climatic conditions.     

The effect of elevated CO2 and temperature on the secondary chemistry of Betula pendula seedlings

The effect of elevated atmospheric CO₂ and temperature on resource allocation and secondary chemistry of white birch (Betula pendula Roth) under a non-limiting nutrient and water supply was investigated. Birch seedlings were grown in closed-top chambers exposed to ambient CO₂ and temperature, elevated atmospheric CO₂ (700 ppm), elevated temperature (2°C above ambient) and a combination of elevated CO₂ and temperature for one growing season. Elevated CO₂ significantly increased the total biomass of the seedlings. The combined effect of the elevated CO₂ and temperature treatments further increased the total biomass, but not significantly. The content of nitrogen and water decreased, while some secondary compounds (such as condensed tannins and flavonol glycosides) increased in leaves subjected to CO₂ enrichment. Elevated temperature increased the concentration of total flavone aglycones and decreased that of total HPLC-phenolics in the leaves, due to the decrease in individual flavonol glycosides, cinnamoylquinic acids and (+)-catechin. There were no significant interactive effects between CO₂ and temperature in the phenolic concentrations of the leaves and in the stems, while the number of resin droplets in the top part of the stems showed significant interaction. This clearly implies that carbon allocation into secondary metabolites in the leaves and stems differ under enhanced CO₂ and temperature, and the combined effect of CO₂ and temperature on the herbivore resistance of birches, is lower than that of CO₂ and temperature alone.     

The effect of heterotrophy on photosynthesis and tissue composition of two scleractinian corals under elevated temperature

Exogenous food can increase protein levels of coral host tissue, zooxanthellae densities, chlorophyll (chl) concentrations and rates of photosynthesis and is thought to play an important role in the resilience of bleached corals. There is however no information about the effect of heterotrophy on the bleaching susceptibility of corals under elevated temperature conditions. This study investigates potential interactions between food availability, basal metabolic functions (photosynthesis and respiration), energy status (lipid concentrations), total protein concentrations and the bleaching susceptibility (loss of chl and/or zooxanthellae) of the scleractinian corals Stylophora pistillata (Esper) and Galaxea fascicularis (Linnaeus) in response to elevated temperature (daily temperature rises of 3-4 °C) over 15 days. Feeding experiments were carried out in which the corals were either fed daily with zooplankton or starved. Compared to fed corals, starvation of both species resulted in a significant decrease in daily photosynthetic oxygen evolution over time. Gross (Pg) and net (Pn) photosynthetic production of starved corals of both species between 10:00-11:00 hrs had declined by ~50% at day 15 while there were no marked changes in Pg and Pn of fed corals. After 15 days, starved S. pistillata contained significantly lower zooxanthellae densities, lipid and protein concentrations than fed corals. Starved G. fascicularis also displayed a decrease in zooxantllae densities which was accompanied by a significant decline in algal chl concentrations. Contrary to S. pistillata, feeding treatment had no effect on the lipid concentrations of G. fascicularis. Total protein concentrations however were significantly lower in straved than in fed G. fascicularis. Furthermore, starvation resulted in a significant decrease in respiration of S. pistillata during the last four days of the experiment while treatment had no effect on the respiration rates of G. fascicularis. Overall the oxygen consumption of S. pistillata of both treatments was about 39-67% higher than the respiration of G. fascicularis indicating that low metabolic rates may have allowed starved G. fascicularis to conserve energy reserves over the course of the experiment. The combined results reveal a strong positive relationship between food availability, sustained photosynthetic activity and reduced loss in pigmentation of both species under elevated temperature conditions.     

The effect of temperature on leaf decomposition and diversity of associated aquatic hyphomycetes depends on the substrate

We examine the relative importance of substrate quality and temperature in the establishment of aquatic hyphomycete assemblages and in their ability to decompose leaves. We used leaves of alder (Alnus glutinosa) and oak (Quercus robur) and we tested four temperatures (5°, 10°, 15° and 20 °C). Differences in decomposition rates and fungal assemblages were higher substrata than across temperatures. In both species, decomposition efficiency measured as the ratio of decay rate to fungal biomass, was greater at higher temperatures. Oak leaves were colonized by fewer aquatic hyphomycete species than was alder. Decomposition rates of oak increased with temperature but that of alder was not affected. We conclude that the substratum is a key driver of aquatic hyphomycete assemblages and can attenuate the effects of temperature differences on litter decomposition.     

The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch

There are reports showing interactive effect of environmental factors with the toxic outcome of chemicals. We studied the interactive effect of elevated temperature as an abiotic stressor on deltamethrin-induced biochemical stress responses in a freshwater fish, Channa punctata Bloch. Heat stress (∼12 °C above ambient temperature for 3 h) and pesticide exposure (deltamethrin 0.75 ppb for 48 h) showed significant induction of heat shock protein-70 (HSP70) in liver, kidney and gills of fishes. Elevated temperature when followed by deltamethrin exposure showed synergistic effect showing a high level of HSP70 in liver and gills whereas response in the kidney was opposite. On the contrary, when deltamethrin exposure followed the heat stress, no significant difference was observed. Protein carbonylation was found to be more pronounced in heat-stressed group compared with control fish group. A significant increase in lipid peroxidation (LPO) was observed in different tissues of fish exposed to either of the stressors. In the kidney of fish exposed to heat stress followed by deltamethrin, LPO was relatively lower as compared to other treatments. Thiols content such as reduced glutathione (GSH), total thiols (T-SH), non-protein thiols (NP-SH) and protein thiols (P-SH) showed no consistent pattern in different tissues. In deltamethrin-exposed group that was subsequently exposed to heat stress, the GSH content was higher in liver and lower in both kidney and gills when compared with other groups. Alteration in the activities of antioxidant enzymes such as catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) was also observed when fish were exposed to heat stress and/or deltamethrin. Our study demonstrated that heat stress modulated biochemical stress responses in fish showing a tissue specific pattern. This implies that fish has the capacity to elicit differential response to exposure to abiotic stressors in order to reduce the systemic magnitude of stress which may otherwise lead to severe dysfunction of vital tissues.     

Stimulation of embryogenesis and haploid production in Brassica campestris anther cultures by elevated temperature treatments

Summary Culture of Brassica campestris anthers at 35°C for one or three days prior to culture at 25°C significantly stimulated the yield of microspore-derived embryos. More than 100 plants were regenerated from cultured embryos and haploids were identified amongst them. The haploid frequency was greater than 70% if all small-flowered sterile plants were considered to be haploid. The yield of microspore-derived plants in B. campestris is approaching the level where anther culture may be utilized as a practical breeding tool.     

Influence of temperature and substrate concentration on bacterial growth yield in Seine River water batch cultures.

The growth of natural bacterial assemblages was monitored in 3-liter reactors under various temperature and substrate concentration conditions. The oxygen concentration was continuously monitored, and subsamples were taken at short time intervals to determine bacterial number and biomass. The rate of bacterial mortality was also determined. Bacterial growth yield was calculated as the ratio of net production (increase in biomass corrected for mortality) to gross production (net production plus oxygen consumption expressed in carbon units). Averaging 33%, the growth yield did not show any trend with temperatures in the range of 8 to 25 degrees C but increased with the concentration of dissolved organic carbon in the range of 2 to 12 mg of C.liter-1.     

The effect of elevated potassium on the time course of neuron survival in cultures of dissociated dorsal root ganglia

The effect of potassium (K⁺) on the time course of neuron survival has been investigated by counting neurons over a 24-day period in live cultures of dissociated dorsal root ganglia from embryonic chick, fetal and newborn mouse, and fetal human material. In both normal K (6 mM) and in elevated K (20 mM mouse and human, 40 mM chick) there was initially a rapid exponential decrease in neuron survival. However, the magnitude of this decrease was less in the elevated K. In normal K neuron number decreased monotonically; the rate of degeneration itself decreased with time so that after 24 days neuron survival became relatively constant. In contrast, in elevated K the neuron number actually increased over a limited time interval before attaining a stable long-term value much greater than that in normal K. Thus, elevated K enhanced long-term survival by causing a lower rate of degeneration and also by causing an increase in neuron number during a limited period of the time in culture. From these observations and other evidence, it is argued that K can substitute to some extent for the trophic action normally exerted by the peripheral field of innervation of the DRG. It is further argued that K acts through its depolarizing effect on the membrane potential and that modification of intracellular ionic concentrations seems less likely to be involved.     

The biosynthesis of nitrogenase MoFe protein polypeptides in free-living cultures of Rhizobium japonicum

The biosynthesis of the constituent polypeptides of nitrogenase component I (Rj 1) in free-living cultures of Rhizobium japonicum (strain 110) was investigated under different growth conditions. Cells were pulse-labelled and the proteins analysed by one and two-dimensional gel electrophoresis. The positions of the constituent Rj 1 polypeptides were identified by co-electrophoresis with purified Rj 1 isolated from bacteroids of soybean nodules, and by comparison with an immunoprecipitate from a culture induced for nitrogenase. The synthesis of the proteins preceded any detectable enzyme activity and increased with time, reaching a maximum after 3 days. At this time, between 6 and 8% of the total sodium dodecyl sulfate-soluble protein synthesized was Rj 1. Exposure to air led to a dramatic decrease in the rate of Rj 1 synthesis, with almost complete regression after 20 min. In the presence of KNO3, there was no nitrogenase activity, but the proteins were present in similar amounts (7%) as the control culture. When mannitol and glycerol were used as the sole carbon sources, the amount of Rj 1 synthesized was extremely low.