Conditions for Maximum Enflagellation in Naegleria fowleri1

ABSTRACT. Ameba to flagellate transformation in Naegleria fowleri (Lovell strain) was affected by growth temperature, phase of growth, strain of ameba, culture agitation, enflagellation temperature, enflagellation diluent, and cell concentration. Amebae transformed best when they were grown without agitation and enflagellated with agitation. Regardless of growth temperature (23°, 30°, 37°, and 42°C were tested), amebae transformed best at 37°C. Enflagellation was greatest for cells harvested between 24 h (mid-exponential) and 84 h (late stationary) of growth.     

Effects of temperature and growth phase on lipid and biochemical composition of Isochrysis galbana TK1

The lipid and biochemical composition of the haptophyte Isochrysis galbana TK1 was examined. Cultures were grown at 15 °C and 30 °C, and harvested in the exponential and early stationary growth phases. Carbohydrate and protein content varied at the two culture temperatures and growth phases. The highest protein content was found at the exponential growth phase at 15 °C, and the highest carbohydrate content was found at the stationary phase at the same culture temperature. Lipid accumulated in the stationary growth phase and its content was higher at 30 °C than at 15 °C regardless of the growth phase. The neutral lipids were the major class of lipid found in all the cultures. The stationary phase culture had a higher proportion of neutral lipids than the exponential phase culture and the proportion decreased slightly when culture temperature was increased from 15 °C to 30 °C. Phospholipid levels remained constant at the two temperatures, but slightly decreased in the stationary phase. Glycolipids in the exponentially growing cells were higher than those from stationary growth phase and increased with temperature. Polyunsaturated fatty acids (PUFAs) predominated in glycolipids and phospholipids. Cells grown at 15 °C contained higher proportion of 18:3 (n–3) and 22:6 (n–3) with a corresponding decrease in 18:2 (n–6), monounsaturated and saturated fatty acids. This revised version was published online in August 2006 with corrections to the Cover Date.     

Fatty acids profile and temperature in the cultured marine diatom Odontella aurita

The diatom Odontella aurita has now been industrially cultured and commercialized as a dietary supplement rich in omega-3 fatty acids for several years. In this study, we investigated the effect of three temperatures (8, 16, and 24 °C) on the growth and fatty acid composition of cells harvested during the exponential and stationary growth phases. These temperatures were selected on the basis of photosynthesis responses previously obtained at different temperatures using a modulated fluorometer. Our results confirm that both growth and lipid composition were sensitive to culture temperature. Growth was reduced when O. aurita was cultured at low temperature (8 °C) compared to when it was cultured at high temperatures (16 and 24 °C), but the proportion of polyunsaturated fatty acids (PUFAs, 20:5 n-3 and 22:6 n-3) increased while the level of saturated fatty acids (SFAs, 14:0 and 16:0) decreased in the cells harvested during both the exponential and stationary growth phases. On the other hand, the cells grown at 24 °C displayed a marked decrease in PUFA and an increase in SFA levels. Harvesting time is also a critical parameter in achieving optimum n-3 PUFA productivity during batch cultivation. Indeed, changes in fatty acid composition with growth phase seem to be dependent on the culture temperature, with the most marked effects being observed at 24 °C. PUFA levels (i.e., levels of 20:5 n-3 and 22:6 n-3) increased during the stationary growth phase, while the proportion of SFAs and monounsaturated fatty acids (MUFAs) fell with time. As this species is currently grown in outdoor ponds with seasonal temperature variations (minimal and maximal average temperatures in winter and summer, from 3 to 9 °C and from 13 to 26 °C, respectively), this factor can be expected to have a strong influence on the fatty acid content and composition of the algal biomass harvested and commercialized.     

Sporulation and synthesis of extracellular proteinases inBacillus subtilis are more temperature-sensitive than growth

Bacillus subtilis 115 grew in a medium with amino acids and glucose with the maximum specific growth rates μ of 1.20-1.10/h in the temperature range of 45–48°C. Activity of the extracellular neutral proteinase excreted by 1.3 mg/mL dry mass during 8 h of the postexponential and stationary growth phases decreased from its maximum value of 0.23 TU/mL at 40°C to 0.13 and 0.06 TU/mL at 45 and 48°C, respectively. Formation of the extracellular serine proteinase decreased even more—from 0.18 TU/mL at 40°C to 0.06 and 0.03 TU/mL at 45 at 48°C, respectively. Sporulation, expressed as the portion of sporangia rith refractile spores at the 6th h of the stationary phase decreased from 46% at 40°C to 17 and 3% at 45 and 48°C, respectively.     

Temperature-induced modifications of glycosphingolipids in plasma membranes of Neurospora crassa

Plasma membranes isolated from a cell-wall-less mutant of Neurospora crassa grown at 37 and 15°C display large differences in lipid compositions. A free sterol-to-phospholipid ratio of 0.8 was found in 37°C membranes, while 15°C plasma membranes exhibited a ratio of nearly 2.0. Membranes formed under both growth conditions were found to contain glycosphingolipids. Cultures grown at the low temperature, however, were found to contain 6-fold higher levels of glycosphingolipids and a corresponding 2-fold reduction of phospholipid levels. The high glycosphingolipid content at 15°C compensates for the reduced levels of phospholipids in such a way that sterol/polar lipid ratios are almost the same in plasma membranes under the two growth conditions. Temperature-dependent changes in plasma-membrane phospholipid and glycosphingolipid species were also observed. Phosphatidylethanolamine levels were sharply reduced at 15°C, in addition to a moderate increase in levels of unsaturated phospholipid fatty acids. Glycosphingolipids contained high levels of long-chain hydroxy fatty acids, which constituted 75% of the total fraction at 37°C, but only 50% at 15°C. Compositional changes were also observed in the long-chain base component of glycosphingolipids with respect to growth temperature. Fluorescence polarization studies indicate that the observed lipid modifications in 15°C plasma membranes act to modulate bulk fluidity of the plasma-membrane lipids with respect to growth temperature. These studies suggest that coordinate modulation of glycosphingolipid, phospholipid and sterol content may be involved in regulation of plasma-membrane fluid properties during temperature acclimation.     

利用原生质体紫外诱变技术选育耐高温香菇菌株

【目的】运用原生质体紫外诱变技术选育香菇耐高温新菌株。【方法】以香菇菌株18为出发菌株,紫外诱变处理其原生质体,通过47°C热激3 h后菌丝恢复生长的情况来筛选获得耐高温诱变株,测定18及其所有诱变株在木屑培养基中的恒温长速、高温长速以及恢复长速,并进行高温出菇试验。【结果】筛选得到57株耐高温诱变株,其中诱变株N6、N44和N24的综合性状较好。恒温长速、高温长速以及恢复长速与出菇性状具有相关性,恢复长速与出菇产量、单菇性状、耐高温能力呈正相关,可初步作为预测耐高温菌株综合性状的指标。【结论】利用原生质体紫外诱变技术,可初步选育出耐高温香菇新菌株。     

The involvement of the lipid phase transition in the plasma-induced dissolution of multilamellar phosphatidylcholine vesicles

Unsonicated liposomes prepared from dimyristoyl phosphatidylcholine were nearly completely dissolved during a 3 h incubation with rat plasma at or close to the phase-transition temperature of 24°C. At 37 or 15°C virtually no liposomal disintegration was observed even after 24 h of incubation. The liposomal solubilization, which was monitored by turbidity measurements or by determination of phospholipid sedimentability, was accompanied by the formation of a phospholipid-protein complex similar or identical to the one we previously reported to be formed from sonicated liposomes of egg phosphatidylcholine (Scherphof, G., Roerdink, F., Waite, M. and Parks, J. (1978) Biochim. Biophys. Acta 542, 296–307). Unsonicated multilamellar liposomes made of egg phosphatidylcholine were completely resistant to the dissolving potency of plasma when incubated at 37°C. Liposomes from equimolar mixtures of dimyristoyl and dipalmitoyl phosphatidylcholine were only degraded by plasma in the temperature range between 30 and 35°C at which temperature this cocrystallizing phospholipid mixture undergoes a phase transition. However, even at these temperatures the rate of dissolution of this mixture was significantly lower than of dimyristoyl phosphatidylcholine at 24°C. In the dissolving process of this mixture a slight preference for the lower-melting component was observed.The ability of cholesterol to completely abolish the susceptibility of dimyristoyl phosphatidylcholine liposomes to plasma at a 1:2 molar ratio of cholesterol to phospholipid substantiates the essential role of the phase transition in the process of liposome solubilization.When liposomes of the monotectic mixtures dimyristoyl and distearoyl phosphatidylcholine or dilauroyl and distearoyl phosphatidylcholine were incubated with plasma at temperatures in between those at which the constituent lipids undergo a phase change in the mixture, the liposomes were slowly disolved. Under those conditions a selective removal of the lipids in the liquid-crystalline phase was observed.It is concluded that for the plasma-induced dissolution of unsonicated liposomes, which is most probably achieved by interaction with (apo)lipoproteins, the presence of phase boundaries is required in much the same way as was first reported for phospholipases by Op den Kamp, J.A.F., de Gier, J. and Van Deenen, L.L.M. (1974) Biochim. Biophys. Acta 345, 253–256).     

The impact of temperature on the production and fitness of microsclerotia of the fungal bioherbicide Mycoleptodiscus terrestris

The impact of growth temperature was evaluated for the fungal plant pathogen Mycoleptodiscus terrestris over a range of temperatures (20–36°C). The effect of temperature on biomass accumulation, colony forming units (cfu), and microsclerotia production was determined. Culture temperatures of 24–30°C produced significantly higher biomass accumulations and 20–24°C resulted in a significantly higher cfu. The growth of M. terrestris was greatly reduced at temperatures above 30°C and was absent at 36°C. The highest microsclerotia concentrations were produced over a wide range of temperatures (20–30°C). These data suggest that a growth temperature of 24°C would optimize the parameters evaluated in this study. In addition to growth parameters, we also evaluated the desiccation tolerance and storage stability of air-dried microsclerotial preparations from these cultures during storage at 4°C. During 5 months storage, there was no significant difference in viability for air-dried microsclerotial preparations from cultures grown at 20–30°C (>72% hyphal germination) or in conidia production (sporogenic germination) for air-dried preparations from cultures grown at 20–32°C. When the effect of temperature on germination by air-dried microsclerotial preparations was evaluated, data showed that temperatures of 22–30°C were optimal for hyphal and sporogenic germination. Air-dried microsclerotial preparations did not germinate hyphally at 36°C or sporogenically at 20, 32, 34, or 36°C. These data show that temperature does impact the growth and germination of M. terrestris and suggest that water temperature may be a critical environmental consideration for the application of air-dried M. terrestris preparations for use in controlling hydrilla.     

Lipid phase separations and intramembranous particle movements in the yeast tonoplast

The tonoplast of Saccharomyces cerevisiae contains regions depleted of intramembranous particles as the cells enter stationary phase. Freeze-fracture studies on intact cells from this growth stage show that a dispersed particle distribution predominates if the cell temperature is raised to 40°C but that particle-depleted areas prevail at or below the cell growth temperature of 30°C. Tonoplasts of isolated vacuoles also contain particle-depleted regions. Differential thermal analyses of lipids extracted from isolated vacuoles show an endothermic transition which encompasses the cell growth temperature. These results suggest that the tonoplast at this stage contains patches of gel-phase lipid and that these patches correspond to the intramembranous particle-depleted areas of the freeze-fractured tonoplast.     

The effect of root temperature on growth of curled parsley

Summary Curled parsley was grown at root-zone temperature (RZT) of 18, 21, 24, 27 and 36°C at air temperature (AT) of 18 and 21°C. Maximum growth was obtained at 18°C AT and 24°C RZT, but there were no significant differences between 18 and 27°C RZT. Shoot and root growth were severely inhibited at 36°C constant RZT. The growth was also retarded when RZT rose to 36°C for 30 minutes per day, even when compared to a RZT of constant 27°C. This indicates that a short exposure to RZT above 30°C retards growth. A relatively low daily average RZT did not compensate for the damage caused by a short daily high temperature exposure. Optimum temperature for curled parsley seems to be about 21°C. Report No. 316.     

Growth and Dormancy in Norway Spruce Ecotypes (Picea abies) I. Interaction of Photoperiod and Temperature

Growth and dormancy as affected by photoperiod and temperature have been studied in Norway spruce ecotypes of different latitudinal and altitudinal origin. First-year seedlings were used. In all ecotypes apical growth cessation and terminal bud formation occurred within 2 weeks after exposure to SD at temperatures of 18 to 24°C. At lower temperatures or at near-critical photoperiods the response was delayed. The critical photoperiod for apical growth cessation varied from 21 hours in ecotype Steinkjer, Norway (64°N) to about 15 hours in ecotype Lankowitz, Austria (47°04′N). High-elevation ecotypes also had longer critical pholoperiods than low-elevation ecotypes from the same latitude. A detectable growth depression resulted from as little as 1 or 2 SDs of 10 hours, and with 4 or more SDs apical growth cessation took place. In contrast to the situation in the shoot, root growth was not affected by photoperiod. Accordingly, the top:root ratio is drastically affected by photoperiod. The critical photoperiod for cambial growth was shorter than that for apical growth in all ecotypes and cambial growth cessation was delayed for several weeks compared with cessation of apical growth. A transition to formation of late-wood tracheids with thick walls and narrow lumens took place upon exposure to SD. The photoperiodic effects were significantly modified by temperature, but the critical photoperiods were only slightly changed by temperature in the range of 12 to 24°C. However, a 10-hour “night” at 4°C caused growth cessation in continuous light in four ecotypes tested. Temperature optimum for apical growth under non-limiting photoperiods (24 hours) was 21°C in all ecotypes, but with little difference among 18,21 and 24°C. The Q₁₀ for apical growth was 3.5 in the temperature range 12 to 18°C. The growth potential as determined in 24-hour photoperiods was not significantly different among the various ecotypes except for one northern eco-type which was clearly inferior to the others. However, the growth of ecotype Steinkjer (64°N) was greatly suppressed even by the long midsummer days at 59°40′N, thus demonstrating the misleading impression one gets of the growth potential of northern ecotypes when they are moved southwards.     

The effect of cessation of growth on protein synthesis in a mutant of Chinese hamster ovary cells with a temperature-sensitive leucyl-tRNA synthetase

A temperature-sensitive mutant of Chinese hamster ovary cells with an altered leucyl-tRNA synthetase fails to grow and to incorporate amino acids into protein properly at or near the non-permissive temperature. This mutant was used to determine whether cessation of growth at the elevated temperature affected elongation factor EF-1, since the activity of EF-1 is markedly lower in non-growing cells in stationary phase than in rapidly-growing cells in exponential phase. Cell-free extracts prepared from cells maintained at 39°C for 24 h showed a marked decrease in the ability to translate natural mRNAs, compared to cells incubated at 34°C. However, the ability to translate poly(U), which requires elongation factor EF-1 (and EF-2), was not affected. Analyses of activities involved in the initiation of protein synthesis and in the activation of amino acids revealed that, with the exception of leucyl-tRNA synthetase, the rest of the components required for translation also appeared to be relatively stable even after 24 h at the elevated temperature. The effects of elevated temperature on cell-free extracts were also investigated. The results were similar to those obtained with intact cells; that is, except for leucyl-tRNA synthetase which was rapidly inactivated in vitro at 39°C, other aminoacyl-tRNA synthetases and translational components involved in chain initiation and elongation were relatively stable. Thus, no change in EF-1 activity was detected as a result of arrested cell growth, an inherent lability of the elongation factor, or metabolic degradation as a consequence of a rapid turnover rate in the absence of protein synthesis.     

Genetic and physiological alterations occurring in a yeast population continuously propagated at increasing temperatures with cell recycling

This work investigated the effects of increasing temperature from 30°C to 47°C on the physiological and genetic characteristics of Saccharomyces cerevisiae strain 63M after continuous fermentation with cell recycling in a system of five reactors in series. Steady state was attained at 30°C, and then the temperature of the system was raised so it ranged from 35°C in the last reactor to 43°C in the first reactor or feeding reactor with a 2°C difference between reactors. After 15 days at steady state, the temperature was raised from 37°C to 45°C for 25 days at steady state, then from 39°C to 47°C for 20 days at steady state. Starter strain 63M was a hybrid strain constructed to have a MAT a/α, LYS/lys, URA/ura genotype. This hybrid yeast showed vigorous growth on plates at 40°C, weak growth at 41°C, positive assimilation of melibiose, positive fermentation of galactose, raffinose and sucrose. Of 156 isolates obtained from this system at the end of the fermentation process, only 17.3% showed the same characteristics as starter strain 63M. Alterations in mating type reaction and in utilization of raffinose, melibiose, and sucrose were identified. Only 1.9% of the isolates lost the ability to grow at 40°C. Isolates showing requirements for lysine and uracil were also obtained. In addition, cell survival was observed at 39–47°C, but no isolates showing growth above 41°C were obtained.     

The influence of the thermal environment and other early life events on growth rate of piglets during lactation

The effects of early life events on average daily weight gain from birth to day 21 (ADG) of suckling pigs kept at different room temperatures (15°C, 20°C and 25°C) from birth to weaning were investigated. Data were collected from litters born by 61 sows in a loose housing system. The ADG for piglets with low birth weight (estimated for birth weights below the 10% percentile) was estimated to be 20 to 30 g higher per day at room temperature 20°C to 25°C compared with 15°C. In contrast, the ADG during the lactation period decreased for larger piglets (estimated for birth weights above the 10% percentile) by 28 g/day at room temperature 25°C compared with 15°C. Thus, high ambient temperatures (20°C to 25°C) are favourable for the growth in smaller piglets during lactation. Neither latency to first suckle nor birth-induced hypoxia, measured as concentration of umbilical cord lactate, affected the growth rate of the piglets. Lowest rectal temperature during the first 24 h after birth had a long-term negative effect on ADG (P<0.05), so that piglets with a lowest rectal temperature of 32.8°C (10% percentile) had an ADG which was on average 19 g lower per day than piglets with a rectal temperature of 37.3°C (90% percentile). Our results showed that hypothermia at birth, low birth weight and high number of suckling piglets lead to reduced ADG during the suckling period. The results suggest that keeping the room temperature at 20°C during lactation to some extent could compensate for the otherwise negative effects of low birth weight on ADG in piglets without decreasing the ADG of high birth weight piglets. However, to avoid hypothermia in the smallest piglets it may be beneficial to increase the room temperature above 20°C during the farrowing period of loose housed sows.     

Reproduction and population parameters of the Nearctic predator Geocoris punctipes at constant and varying temperature regimes

The heteropteran predator Geocoris punctipes (Say) has been used in augmentative biological control since 2000 to control Lepidoptera. However, surprisingly, few data are available about the influence of temperature on its population development, which is of key importance to plan the number and moment of releases to obtain sufficient pest reduction. The objective of this study was to evaluate daily and total fecundity, longevity and life table parameters (m_x, l_x, r_m, R, λ, T and TD) of G. punctipes at constant (16.8°C, 21.5°C, 24.5°C and 28.3°C) and corresponding varying (day/night) (21/11°C, 24/18°C, 27/21°C and 30/26°C) temperatures. Pairs of adult predators aged 24 h and originating from nymphs exposed to the same temperature regimes were kept at the above‐mentioned temperature regimes in Petri dishes containing Anagasta kuehniella (Zeller) eggs and an oviposition substrate. Tests were conducted in climatic chambers at the different temperature regimes and a RH 70 ± 10% and a 14L: 10D photoperiod. Reproduction, longevity and life table parameters were significantly affected by temperature, with clear differences between treatments at low (16.8°C, 21/11°C, 21.5°C, 24/18°C) or a high (24.5°C, 27/21°C, 28.3°C, 30/26°C) temperature regimes. Highest reproduction and fastest population growth of G. punctipes took place at average temperatures ranging from 24.5°C to 30°C, and neither reproduction nor population growth was negatively influenced by varying temperatures at any of the temperature regimes.     

Temperature dependence of d-glucose transport in reconstituted liposomes

Sodium-dependent d-glucose uptake into proteoliposomes reconstituted from dimyristoylphosphatidylcholine (DMPC) and hog kidney brush border membrane extract is strongly affected by temperature and the physical state of the membranes. This dependence is defined by a nonlinear Arrhenius plot with a break point at 23°C, a temperature not significantly different from the phase transition temperature of the pure lipid (24°C). The transport process is characterized by different activation energies: 35.1 kcal/mol below and 5.5 kcal/mol above the transition temperature. The shift in the break point for the d-glucose transport activity from 15°C, in the brush border membranes, to 23°C in the reconstituted system leads us to conclude that the lipids surrounding the sodium/d-glucose cotransport system can exchange readily with the bulk lipid used for reconstitution. The results thus provide no evidence for the presence of an annulus of specific lipids surrounding the transport system.     

CHARACTERIZATION OF PSYCHROPHILIC OSCILLATORIANS (CYANOBACTERIA) FROM ANTARCTIC MELTWATER PONDS

Oscillatorian cyanobacteria dominate benthic microbial mat communities in many polar freshwater ecosystems. Capable of growth at low temperatures, all benthic polar oscillatorians characterized to date are psychrotolerant (growth optima > 15° C) as opposed to psychrophilic (growth optima ≤ 15° C). Here, psychrophilic oscillatorians isolated from meltwater ponds on Antarctica's McMurdo Ice Shelf are described. Growth and photosynthetic rates were investigated at multiple temperatures, and compared with those of a psychrotolerant isolate from the same region. Two isolates showed a growth maximum at 8° C, with rates of 0.12 and 0.08 doublings·d ^{? 1}, respectively. Neither displayed detectable growth at 24° C. The psychrotolerant isolate showed almost imperceptible growth at 4° C and a rate of 0.9 doublings·d ^{? 1} at its optimal temperature of ～23° C. In both photosynthesis versus irradiance and photosynthesis versus temperature experiments, exponentially growing cultures were acclimated for 14 days at 3, 8, 12, 20, and 24° C under saturating light intensity, and [¹⁴C] photoincorporation rates were measured. Psychrophilic isolates acclimated at 8° C showed greatest photosynthetic rates; those acclimated at 3° C were capable of active photosynthesis, but photoincorporation was not detected in cells acclimated at 20 and 24° C, because these isolates were not viable after 14 days at those temperatures. The psychrotolerant isolate, conversely, displayed maximum photosynthetic rates at 24° C, though photoincorporation was actively occurring at 3° C. Within acclimation temperature treatments, short‐term photosynthetic rates increased with increasing incubation temperature for both psychrophilic and psychrotolerant isolates. These results indicate the importance of temperature acclimation before assays when determining optimal physiological temperatures. All isolates displayed photosynthetic saturation at low light levels (<128 μmol·m ^{? 2}·s ^{? 1}) but were not photoinhibited at the highest light treatment (233 μmol·m ^{? 2}·s ^{? 1}). Field studies examining the impact of temperature on photosynthetic responses of intact benthic mats, under natural solar irradiance, showed the mat communities to be actively photosynthesizing from 2 to 20° C, with maximum photoincorporation at 20° C, as well as capable of a rapid response to an increase in temperature. The rarity of psychrophilic cyanobacteria, relative to psychrotolerant strains, may be due to their extremely slow growth rates and inability to take advantage of occasional excursions to higher temperatures. We suggest an evolutionary scenario in which psychrophilic strains, or their most recent common ancestor, lost the ability to grow at higher temperatures while maintaining a broad tolerance for fluctuations in other physical and chemical parameters that define shallow meltwater Antarctic ecosystems.