Seasonal changes in energetics and torpor patterns in the subtropical blossom-bat Syconycteris australis (Megachiroptera)

Little is known about how animals from tropical and subtropical climates adjust their energy expenditure to cope with seasonal changes of climate and food availability. To provide such information, we studied the thermal physiology, torpor patterns and energetics of the nocturnal blossom-bat (Syconycteris australis 18 g) from a subtropical habitat in both summer and winter. In both seasons, S. australis frequently entered daily torpor at ambient temperatures between 12 and 25°C when food and water were withheld. Unlike patterns observed in temperate animals, mean minimum metabolic rates during torpor were lower in summer (0.47 ± 0.07 ml O₂ g⁻¹ h⁻¹) than in winter (0.75 ± 0.11 ml O₂ g⁻¹ h⁻¹). Body temperatures during torpor were regulated at 19.3 ± 1.0°C in summer and at 23.4 ± 2.0°C in winter. Torpor bout duration was significantly longer in summer (7.3 ± 0.6 h) than in winter (5.5 ± 0.3 h), but in both seasons, bout duration was not affected by ambient temperature. Consequently, average daily metabolic rates were also significantly lower in summer than in winter. Body temperatures and metabolic rates in normothermic bats did not change with season. Our findings on seasonal changes of torpor in this bat from the subtropics are opposite to those made for many species from cold climates which generally show deeper and longer torpor in winter and are often entirely homeothermic in summer. More pronounced torpor in subtropical S. australis in summer may be due to low or unpredictable nectar availability, short nights which limit the time available for foraging, and long days without access to food. Thus, the reversed seasonal response of this subtropical bat in comparison to temperate species may be an appropriate response to ecological constraints. Received: 6 May 1997 / Accepted: 19 October 1997     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000     

Accurate determination of the oxidative phosphorylation affinity for ADP in isolated mitochondria

Background

Mitochondrial dysfunctions appear strongly implicated in a wide range of pathologies. Therefore, there is a growing need in the determination of the normal and pathological integrated response of oxidative phosphorylation to cellular ATP demand. The present study intends to address this issue by providing a method to investigate mitochondrial oxidative phosphorylation affinity for ADP in isolated mitochondria.

Methodology/Principal Findings

The proposed method is based on the simultaneous monitoring of substrate oxidation (determined polarographically) and phosphorylation (determined using the glucose - hexokinase - glucose-6-phosphate dehydrogenase - NADP⁺ enzymatic system) rates, coupled to the determination of actual ADP and ATP concentrations by bioluminescent assay. This enzymatic system allows the study of oxidative phosphorylation during true steady states in a wide range of ADP concentrations. We demonstrate how the application of this method allows an accurate determination of mitochondrial affinity for ADP from both oxidation (K_mVox) and phosphorylation (K_mVp) rates. We also demonstrate that determination of K_mVox leads to an important overestimation of the mitochondrial affinity for ADP, indicating that mitochondrial affinity for ADP should be determined using phosphorylation rate. Finally, we show how this method allows the direct and precise determination of the mitochondrial coupling efficiency. Data obtained from rat skeletal muscle and liver mitochondria illustrate the discriminating capabilities of this method.

Conclusions/Significance

Because the proposed method allows the accurate determination of mitochondrial oxidative phosphorylation affinity for ADP in isolated mitochondria, it also opens the route to a better understanding of functional consequences of mitochondrial adaptations/dysfunctions arising in various physiological/pathophysiological conditions.     

Purification and Biochemical Characterization of Digestive Lipase in Whiteleg Shrimp

Penaeus vannamei lipase was purified from midgut gland of whiteleg shrimp. Pure lipase (E.C. 3.1.1.3) was obtained after Superdex 200 gel filtration and Resource Q anionic exchange. The pure lipase, which is a glycosylated molecule, is a monomer having a molecular mass of about 44.8 kDa, as determined by SDS-PAGE analysis. The lipase hydrolyses short and long-chain triacylglycerols and naphthol derivates at comparable rates. A specific activity of 1787 U mg⁻¹ and 475 U mg⁻¹ was measured with triolein and tributyrin as substrates, respectively, at pH 8.0 and 30°C in the absence of colipase. The lipase showed a K _{m, app} of 3.22 mM and k _{cat, app}/K _{m, app} of 0.303 × 10³ mM⁻¹ s⁻¹ using triolein as substrate. Natural detergents, such as sodium deoxycholate, act as potent inhibitors of the lipase. This inhibition can be reversed by adding fresh oil emulsion. Result with tetrahydrolipstatin, an irreversible inhibitor, suggests that the lipase is a serine enzyme. Peptide sequences of the lipase were determined and compared with the full-length sequence of lipase which was obtained by the rapid amplification of cDNA ends method. The full cDNA of the pvl was 1,186 bp, with a deduced protein of 362 amino acids that includes a consensus sequence (GXSXG) of the lipase superfamily of α/β-hydrolase. The gene exhibits features of conserved catalytic residues and high homology with various mammalian and insect lipase genes. A potential lid sequence is suggested for pvl.     

Actual and potential rates of hydrogen photoproduction by continuous culture of the purple non-sulphur bacterium Rhodobacter capsulatus

The influence of (NH₄)₂SO₄ concentration and dilution rate (D) on actual and potential H₂ photoproduction has been studied in ammonium-limited chemostat cultures of Rhodobacter capsulatus B10. The actual H₂ production in a photobioreactor was maximal (approx. 80 ml h⁻¹l⁻¹) at D = 0.06 h⁻¹ and 4 mM (NH₄)₂SO₄. However, it was lower than the potential H₂ evolution (calculated from hydrogen evolution rates in incubation vials), which amounted to 100–120 ml h⁻¹ l⁻¹ at D = 0.03–0.08 h⁻¹. Taking into account the fact that H₂ production in the photobioreactor under these conditions was not limited by light or lactate, another limiting (inhibiting) factor should be sought. One possibility is an inhibition of H₂ production by the H₂ accumulated in the gas phase. This is apparent from the non-linear kinetics of H₂ evolution in the vials or from its inhibition by the addition of H₂; initial rates were restored in both cases after the vials had been refilled with argon. The actual H₂ production in the photobioreactor at D = 0.06 h⁻¹ was shown to increase from approximately 80 ml h⁻¹ l⁻¹ to approximately 100 ml h⁻¹ l⁻¹ under an argon flow at 100 ml min⁻¹. Under maximal H₂ production rates in the photobioreactor, up to 30% of the lactate feedstock was utilised for H₂ production and 50% for biomass synthesis. Received: 22 April 1997 / Received revision: 14 July 1997 / Accepted: 27 July 1997     

Effects of 3 days of carbohydrate supplementation on muscle glycogen content and utilisation during a 1-h cycling performance

This study compared the effects of supplementing the normal diets of six trained cyclists [maximal oxygen uptake O_2max) 4.5 (0.36)l · min⁻¹; values are mean (SD)] with additional carbohydrate (CHO) on muscle glycogen utilisation during a 1-h cycle time-trial (TT). Using a randomised crossover design, subjects consumed either their normal diet (NORM) for 3 days, which consisted of 426 (137) g · day⁻¹ CHO [5.9 (1.4) g · kg⁻¹ body mass (BM)], or additional CHO (SUPP) to increase their intake to 661 (76) g · day⁻¹ [9.3 (0.7) g · kg⁻¹ BM]. The SUPP diet elevated muscle glycogen content from 459 (83) to 565 (62) mmol · kg⁻¹ dry weight (d.w.) (P < 0.05). However, despite the increased pre-exercise muscle glycogen stores, there was no difference in the distance cycled during the TT [40.41 (1.44) vs 40.18 (1.76) km for NORM and SUPP, respectively]. With NORM, muscle glycogen declined from 459 (83) to 175 (64) mmol · kg⁻¹ d.w., whereas with SUPP the corresponding values were 565 (62) and 292 (113) mmol · kg⁻¹ d.w. Accordingly, both muscle glycogen utilisation [277 (64) vs 273 (114) mmol · kg⁻¹ d.w.] and total CHO oxidation [169 (20) vs 165 (30) g · h⁻¹ for NORM and SUPP, respectively] were similar. Neither were there any differences in plasma glucose or lactate concentrations during the two experimental trials. Plasma glucose concentration averaged 5.5 (0.5) and 5.6 (0.6) mmol · l⁻¹, while plasma lactate concentration averaged 4.4 (1.9) and 4.4 (2.3) mmol · l⁻¹ for NORM and SUPP, respectively. The results of this study show that when well-trained subjects increase the CHO content of their diet for 3 days from 6 to 9 g · kg⁻¹ BM there is only a modest increase in muscle glycogen content. Since supplementary CHO did not improve TT performance, we conclude that additional CHO provides no benefit to performance for athletes who compete in intense, continuous events lasting 1 h. Furthermore, the substantial muscle CHO reserves observed at the termination of exercise indicate that whole-muscle glycogen depletion does not determine fatigue at this exercise intensity and duration. Accepted: 25 November 1996     

Kinetics of styrene biodegradation by <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. E-93486

The research into kinetics of styrene biodegradation by bacterial strain Pseudomonas sp. E-93486 coming from VTT Culture Collection (Finland) was presented in this work. Microbial growth tests in the presence of styrene as the sole carbon and energy source were performed both in batch and continuous cultures. Batch experiments were conducted for initial concentration of styrene in the liquid phase changed in the range of 5–90 g m⁻³. The Haldane model was found to be the best to fit the kinetic data, and the estimated constants of the equation were: μ _m = 0.1188 h⁻¹, K _S = 5.984 mg l⁻¹, and K _i = 156.6 mg l⁻¹. The yield coefficient mean value Y_\textxs^\textapp Y_{\text{xs}}^{\text{app}} for the batch culture was 0.72 g_{dry cells weight} (g_substrate)⁻¹. The experiments conducted in a chemostat at various dilution rates (D = 0.035–0.1 h⁻¹) made it possible to determine the value of the coefficient for maintenance metabolism m _d = 0.0165 h⁻¹ and the maximum yield coefficient value Y_\textxs^\textM = 0.913 Y_{\text{xs}}^{\text{M}} = 0.913 . Chemostat experiments confirmed the high value of yield coefficient Y_\textxs^\textapp Y_{\text{xs}}^{\text{app}} observed in the batch culture. The conducted experiments showed high activity of the examined strain in the styrene biodegradation process and a relatively low sensitivity to inhibition of its growth at higher concentrations of styrene in the solution. Such exceptional features of Pseudomonas sp. E-93486 make this bacterial strain the perfect candidate for technical applications.     

Determination of the kinetic parameters during continuous cultivation of the lipase-producing thermophile Bacillus sp. IHI-91 on olive oil

A thermostable lipase was produced in continuous cultivation of a newly isolated thermophilic Bacillus sp. strain IHI-91 growing optimally at 65 °C. Lipase activity decreased with increasing dilution rate while lipase productivity showed a maximum of 340 U l⁻¹ h⁻¹ at a dilution rate of 0.4 h⁻¹. Lipase productivity was increased by 50% compared to data from batch fermentations. Up to 70% of the total lipase activity measured was associated to cells and by-products or residual substrate. Kinetic and stoichiometric parameters for the utilisation of olive oil were determined. The maximal biomass output method led to a saturation constant K _S of 0.88 g/l. Both batch growth data and a washout experiment yielded a maximal specific growth rate, μ_max, of 1.0 h⁻¹. Oxygen uptake rates of up to 2.9 g l⁻¹h⁻¹ were calculated and the yield coefficient, Y _X/O, was determined to be 0.29 g dry cell weight/g O₂. From an overall material balance the yield coefficient, Y _X/S, was estimated to be 0.60 g dry cell weight/g olive oil. Received: 8 January 1997 / Received revision: 30 April 1997 / Accepted: 4 May 1997     

Influence of inorganic phosphate and energy state on force in skinned cardiac muscle from freshwater turtle and rainbow trout

Inorganic phosphate, which increases in the hypoxic cardiac cell, depresses force development. The cardiac muscle of freshwater turtle maintains a remarkably high contractility during hypoxia; this may involve a low sensitivity to phosphate. Therefore, freshwater turtle and rainbow trout were compared with regard to Ca²⁺-activated force in skinned atrial trabeculae in a bath containing 3 mM ATP buffered by 15 mM creatine phosphate in the presence of creatine kinase. For turtle, an increase in phosphate from 0 mM to either 6 mM or 12 mM reduced maximal force by 50% and 80% respectively, whereas the Ca²⁺ activity eliciting half maximal force (Ca_0.5) was increased by 70% in 6 mM and could not be reliably recorded in 12 mM. For trout, the effects of phosphate were less pronounced. An increase from 0 mM to 12 mM did not affect maximal force significantly, but elevated Ca_0.5 by 70%. Hypoxia increases ADP as creatine phosphate is shifted to creatine, therefore, creatine phosphate was changed from 15 mM to 3 mM and creatine from 0 mM to 12 mM. After these changes, the elevation of phosphate from 0 mM to 12 mM had no significant effects for either turtle or trout. In conclusion, the high performance of turtle cardiac muscle during hypoxia does not involve a low sensitivity of the contractile system to phosphate. In addition, the effect of increased phosphate seems to be offset by a concomitant increase in ADP. Accepted: 28 June 1999     

Respiratory and metabolic responses of the Australian Yabby Cherax destructor to progressive and sustained environmental hypoxia

The Australian Yabby, Cherax destructor, inhabits occasionally hypoxic water. The respiratory gas, acid-base, metabolite and energetic status of this crayfish was assessed during progressive hypoxia and during 3 h at a water PO₂ of 1.33 kPa. The O₂ affinity of haemocyanin from C. destructor was increased by lactate (Δlog P ₅₀/Δlog[lactate] = −0.111) and by Ca (Δlog P ₅₀/Δlog[Ca] = −0.62) but not by urate. While the non-bicarbonate buffering capacity was low (Δ[HCO₃ ⁻]/ ΔpH=−4.89) the haemocyanin had a low sensitivity to pH changes (ϕ = −0.33). The crayfish showed a compensatory hyperventilation, which induced a respiratory alkalosis, until the water O₂ partial pressure declined below 2.67 kPa, after which the O₂ uptake rate was approximately 10% of normoxic rates. The high haemocyanin-O₂ affinity maintained haemolymph O₂ content during progressive hypoxia despite the normally low arterial O₂ partial pressure of C. destructor. During severe hypoxia, pH decreased but increased lactate aided in maintaining haemocyanin-O₂ saturation. The importance of regulated haemocyanin-O₂ affinity in hypoxic C. destructor was reduced by lowered metabolism, including reduced cardiac output, and the consequent reduction in O₂ requirement. Anaerobiosis became important only at very low PO₂ but thereafter proceeded rapidly, supported by a marked hyperglycaemia. There was no depletion of adenylates, even after 3 h of severe hypoxia. The tail muscle of C. destructor held small amounts of glycogen which would sustain anaerobiosis for a only a few hours. Hypometabolism seems an important hypoxic response but severe hypoxia may encourage the crayfish to breathe air. Accepted: 26 February 1998     

Exchanges of oxygen, carbon dioxide, nitrogen and water in the caecilian Dermophis mexicanus

Oxygen consumption was measured in five Dermophis mexicanus and averaged (±SEM) 0.047 ± 0.004 ml O₂ g⁻¹ h⁻¹. Carbon dioxide production averaged 0.053 ± 0.005 ml CO₂ g⁻¹ h⁻¹ in the same five animals 1 week later. This metabolic rate is similar to metabolic rates of other Gymnophionans but lower than metabolic rates reported for Anurans and Urodeles. Total nitrogen excretion averaged 1.37 μmol N g⁻¹ h⁻¹ which is higher than that found for other amphibians. Of this, 82.5% (1.13 μmol N g⁻¹ h⁻¹) was in the form of urea while 17.5% (0.24 μmol N g⁻¹ h⁻¹) was in the form of NH₃ + NH⁺ ₄. Such ureotelism is typical of terrestrial amphibians like D. mexicanus. Osmotic water flux averaged 0.0193 ml g⁻¹ h⁻¹ in control (sham injected) animals and was not significantly altered by injection of either arginine vasotocin or mesotocin. This osmotic flux is similar to osmotic fluxes found for other terrestrial amphibians. The combined data suggest that metabolism in D. mexicanus is, like most other Gymnophionans, lower than other amphibians. The high rates of nitrogen (especially urea) excretion suggests that this fossorial animal accumulates urea like other burrowing amphibians. Accepted: 27 June 2000     

Failure to obtain a unique threshold on the blood lactate concentration curve during exercise

The purpose of this study was to compare various methods and criteria used to identify the anaerobic threshold (AT), and to correlate the AT obtained with each other and with running performance. Furthermore, a number of additional points throughout the entire range of lactate concentrations [La⁻] were obtained and correlated with performance. A group of 19 runners [mean age 33.7 (SD 9.6) years, height 173 (SD 6.3) cm, body mass 68.3 (SD 5.4) kg, maximal O₂ uptake (V˙O_{2 max} ) 55.2 (SD 5.9) ml · kg⁻¹ · min⁻¹] performed a maximal multistage treadmill test (1 km · h⁻¹ every 3.5 min) with blood sampling at the end of each stage while running. All AT points selected (visual [La⁻], 4 mmol · l⁻¹ [La⁻], 1 mmol · l⁻¹ above baseline, log-log breakpoint, and 45° tangent to the exponential regression) were highly correlated one with another and with performance (r > 0.90) even when there were many differences among the AT (P < 0.05). The additional points (ranging from 3 to 8 mmol · l⁻¹ [La⁻], 1 to 6 mmol · l⁻¹ [La⁻] above the baseline, and 30 to 70° tangent to the exponential curve of [La⁻]) were also highly correlated with performance (r > 0.90). These results failed to demonstrate a distinct AT because many points of the curve provided similar information. Intercorrelations and correlations between AT and performance were, however, reduced when AT were expressed as the percentage of maximal treadmill speed obtained at AT or percentage of V˙O_{2 max} . This would indicate that different attributes of aerobic performance (i.e. maximal aerobic power, running economy and endurance) are measured when manipulating units. Thus, coaches should be aware of these results when they prescribe an intensity for training and concentrate more on the physiological consequences of a chosen [La⁻] rather than on a “threshold”. Accepted: 22 October 1997     

CO2 exchange of the endolithic lichen Verrucaria baldensis from karst habitats in northern Italy

CO₂ exchange of the endolithic lichen Verrucaria baldensis was measured in the laboratory under different conditions of water content, temperature, light, and CO₂ concentration. The species had low CO₂ exchange rates (maximum net photosynthesis: c. 0.45 μmol CO₂ m⁻² s⁻¹; maximum dark respiration: c. 0.3 μmol CO₂ m⁻² s⁻¹) and a very low light compensation point (7 μmol photons m⁻² s⁻¹ at 8°C). The net photosynthesis/respiration quotient reached a maximum at 9–15°C. Photosynthetic activity was affected only after very severe desiccation, when high resaturation respiratory rates were measured. Microclimatic data were recorded under different weather conditions in an abyss of the Trieste Karst (northeast Italy), where the species was particularly abundant. Low photosynthetically active radiation (normally below 40 μmol photons m⁻² s⁻¹), very high humidities (over 80%), and low, constant temperatures were measured. Thallus water contents sufficient for CO₂ assimilation were often measured in the absence of condensation phenomena. Received: 22 September 1996 / Accepted: 26 April 1997     

Hydrogen production with high yield and high evolution rate by self-flocculated cells of Enterobacter aerogenes in a packed-bed reactor

Continuous hydrogen gas evolution by self-flocculated cells of Enterobacter aerogenes, a natural isolate HU-101 and its mutant AY-2, was performed in a packed-bed reactor under glucose-limiting conditions in a minimal medium. The flocs that formed during the continuous culture were retained even when the dilution rate was increased to 0.9 h⁻¹. The H₂ production rate increased linearly with increases in the dilution rate up to 0.67 h⁻¹, giving maximum H₂ production rates of 31 and 58 mmol l⁻¹ h⁻¹ in HU-101 and AY-2 respectively, at a dilution rate of more than 0.67 h⁻¹. The molar H₂ yield from glucose in AY-2 was maintained at about 1.1 at dilution rates between 0.08 h⁻¹ and 0.67 h⁻¹, but it decreased rapidly at dilution rates more than 0.8 h⁻¹. Received: 27 August 1997 / Received revision: 11 November 1997 / Accepted: 14 December 1997     

Intracellular pH of crab and crayfish muscle fibre types during GABA application and inhibitory nerve stimulation

The inhibitory motoneurons of crustaceans form synapses both with the sarcolemma of muscle fibres and with the very distal branchings of the excitatory motoneurons. The transmitter of these synapses is GABA (γ-aminobutyric acid) which is known to open Cl⁻ channels. Studies on the dactyl opener muscle of crayfish suggest that application of GABA not only leads to an increase in the Cl⁻ permeability but also to a considerable HCO⁻ ₃ conductance that causes an intracellular acidification. To investigate possible physiological implications, we measured the intracellular pH of various muscle fibre types of crayfish and crab using pH-sensitive microelectrodes. Independent of the presence or absence of inhibitory innervation, bath application of 10⁻⁵ mol l⁻¹ GABA led to acidification in all fibre types (pH change: 0.14 ± 0.08, n=11). In no preparation was a change in intracellular pH observed upon stimulation of specific or common inhibitory motoneurons with 10–40 pulses s⁻¹ for 2–5 min. The results suggest that HCO⁻ ₃ conductance cannot be activated through synaptic GABA receptors. However, all crustacean muscle fibre types seem to possess extrasynaptic GABA-sensitive channels that exhibit a considerable HCO⁻ ₃ conductance. The physiological importance of these channels remains to be elucidated. Accepted: 13 July 2000     

Relationship between maximal oxygen uptake on different ergometers, lean arm volume and strength in paraplegic subjects

The present experiment was designed to study the importance of strength and muscle mass as factors limiting maximal oxygen uptake (V˙O_{2 max} ) in wheelchair subjects. Thirteen paraplegic subjects [mean age 29.8 (8.7) years] were studied during continuous incremental exercises until exhaustion on an arm-cranking ergometer (AC), a wheelchair ergometer (WE) and motor-driven treadmill (TM). Lean arm volume (LAV) was estimated using an anthropometric method based upon the measurement of various circumferences of the arm and forearm. Maximal strength (MVF) was measured while pushing on the rim of the wheelchair for three positions of the hand on the rim (−30°, 0° and +30°). The results indicate that paraplegic subjects reached a similar V˙O_{2 max} [1.23 (0.34) l · min⁻¹, 1.25 (0.38) l · min⁻¹, 1.22 (0.18) l · min⁻¹ for AC, TM and WE, respectively] and V˙O_{2 max} /body mass [19.7 (5.2) ml · min⁻¹ · kg⁻¹, 19.5 (6.14) ml · min⁻¹ · kg⁻¹, 19.18 (4.27) ml · min⁻¹ · kg⁻¹ for AC, TM and WE, respectively on the three ergometers. Maximal heart rate f _{c max} during the last minute of AC (173 (17) beats · min⁻¹], TM [168 (14) beats · min⁻¹], and WE [165 (16) beats · min⁻¹], were correlated, but f _{c max} was significantly higher for AC than for TM (P<0.03). There were significant correlations between MVF and LAV (P<0.001) and between the MVF data obtained at different angles of the hand on the rim [311.9 (90.1) N, 313.2 (81.2) N, 257.1 (71) N, at −30°, 0° and +30°, respectively]. There was no correlation between V˙O_{2 max} and LAV or MVF. The relatively low values of f _{c max} suggest that V˙O_{2 max} was, at least in part, limited by local aerobic factors instead of central cardiovascular factors. On the other hand, the lack of a significant correlation between V˙O_{2 max} and MVF or muscle mass was not in favour of muscle strength being the main factor limiting V˙O_{2 max} in our subjects. Accepted: 31 January 1997     

Photosynthetic characteristics of dipterocarp seedlings in three tropical rain forest light environments: a basis for niche partitioning?

In three tropical rain forest light environments in Sabah, Malaysia, we compared photosynthesis in seedlings of ten climax tree species with putatively differing shade tolerances. The objectives of the study were (a) to characterise the range of photosynthetic responses in ten species of the Dipterocarpaceae and (b) to elucidate those photosynthetic characteristics that might provide a basis for niche partitioning. Seedlings were acclimated (c. 7 months) in three light environments; understorey, partial shade and a gap (140 m²). The light environments represented a gradation in median diurnal (0630–1830 hours) photon flux density (PFD) ranging from understorey (4.7 μmol m⁻² s⁻¹), through partial shade (21.2 μmol m⁻² s⁻¹) to gap (113.7 μmol m⁻² s⁻¹). Integrated diurnal PFD were in the sequence gap > partial shade > understorey (15.2, 4.7, 1.3 mol m⁻² day⁻¹, respectively). In gap-acclimated plants, species differed in the photosynthetic light-response variables apparent quantum yield, dark respiration rate, light compensation point, net saturated leaf assimilation rate (A _sat), and in stomatal conductance (g _s sat) when assimilation rate (A) was saturated. A light-demanding pioneer species (Macaranga hypoleuca) and a shade-demanding understorey species (Begonia sp.) had, respectively, higher and lower A _sat and g _s sat than the dipterocarp species. In high-light conditions A _sat and g _s sat were strongly positively correlated in dipterocarp species. Differing photosynthetic characteristics of gap-acclimated plants suggest that, in these dipterocarp species, different rates of carbon fixation may be an important factor contributing towards niche partitioning. Mean integrated diurnal A (A _diurnal) in the gap, partial shade and understory were, respectively, 122.9, 52.7, 20.5 mmol m⁻² day⁻¹. Differences occurred in A _diurnal of dipterocarp species between light environments. When Macaranga was included, differences in A _diurnal were evident in the gap and partial shade, and in both cases were attributed to the pioneer. For the variable A _diurnal, there was of a shift in the rank position of Macaranga among light environments, but a shift did not occur among the dipterocarp species. Results from this study are consistent with the idea that rates of carbon fixation per unit leaf area may contribute towards niche differentiation between the climax and single pioneer species, but not within the group of climax species. Other physiological and/or carbon allocation factors may be involved in any niche partitioning; dipterocarp species often have inherently different growth rates and susceptibility to herbivory. As an alternative to niche partitioning, dipterocarp species may co-exist in natural light environments as a result of habitat disequilibrium or purely stochastic processes. Received: 2 April 1997 / Accepted: 13 July 1997     

Effects of temperature on photosynthesis of two morphologically contrasting plant species along an altitudinal gradient in the tropical high Andes

The effects of temperature on photosynthesis of a rosette plant growing at ground level, Acaena cylindrostachya R. et P., and an herb that grows 20–50 cm above ground level, Senecio formosus H.B.K., were studied along an altitudinal gradient in the Venezuelan Andes. These species were chosen in order to determine – in the field and in the laboratory – how differences in leaf temperature, determined by plant form and microenvironmental conditions, affect their photosynthetic capacity. CO₂ assimilation rates (A) for both species decreased with increasing altitude. For Acaena leaves at 2900 m, A reached maximum values above 9 μmol m⁻² s⁻¹, nearly twice as high as maximum A found at 3550 m (5.2) or at 4200 m (3.9). For Senecio leaves, maximum rates of CO₂ uptake were 7.5, 5.8 and 3.6 μmol m⁻² s⁻¹ for plants at 2900, 3550 and 4200 m, respectively. Net photosynthesis-leaf temperature relations showed differences in optimum temperature for photosynthesis (A _o.t.) for both species along the altitudinal gradient. Acaena showed similar A _o.t. for the two lower altitudes, with 19.1°C at 2900 m and 19.6°C at 3550 m, while it increased to 21.7°C at 4200 m. Maximum A for this species at each altitude was similar, between 5.5 and 6.0 μmol m⁻² s⁻¹. For the taller Senecio, A _o.t. was more closely related to air temperatures and decreased from 21.7°C at 2900 m, to 19.7°C at 3550 m and 15.5°C at 4200 m. In this species, maximum A was lower with increasing altitude (from 6.0 at 2900 m to 3.5 μmol m⁻² s⁻¹ at 4200 m). High temperature compensation points for Acaena were similar at the three altitudes, c. 35°C, but varied in Senecio from 37°C at 2900 m, to 39°C at 3550 m and 28°C at 4200 m. Our results show how photosynthetic characteristics change along the altitudinal gradient for two morphologically contrasting species influenced by soil or air temperatures. Received: 5 July 1997 / Accepted: 25 October 1997     

Protozoan growth rates in Antarctic lakes

The growth rates of heterotrophic nanoflagellates (HNAN), mixotrophic cryptophytes, dinoflagellates and ciliates in field assemblages from Ace Lake in the Vestfold Hills (eastern Antarctica) and Lakes Fryxell and Hoare (McMurdo Dry Valleys, western Antarctica), were determined during the austral summers of 1996/1997 and 1997/1998. The response of the nanoflagellates to temperature differed between lakes in eastern and western Antarctica. In Ace Lake the available bacterial food resources had little impact on growth rate, while temperature imposed an impact, whereas in Lake Hoare increased bacterial food resources elicited an increase in growth rate. However, the incorporation of published data from across Antarctica showed that temperature had the greater effect, but that growth is probably controlled by a suite of factors not solely related to bacterial food resources and temperature. Dinoflagellates had relatively high specific growth rates (0.0057–0.384 h⁻¹), which were comparable to Antarctic lake ciliates and to dinoflagellates from warmer, lower latitude locations. Temperature did not appear to impose any significant impact on growth rates. Mixotrophic cryptophytes in Lake Hoare had lower specific growth rates than HNAN (0.0029–0.0059 h⁻¹ and 0.0056–0.0127 h⁻¹, respectively). They showed a marked seasonal variation in growth rate, which was probably related to photosynthetically active radiation under the ice at different depths in the water column. Ciliates' growth rates showed no relationship between food supply and mean cell volume, but did show a response to temperature. Specific growth rates ranged between 0.0033 and 0.150 h⁻¹ for heterotrophic ciliates, 0.0143 h⁻¹ for a mixotrophic Plagiocampa species and 0.0075 h⁻¹ for the entirely autotrophic ciliate, Mesodinium rubrum. The data indicated that the scope for growth among planktonic Protozoa living in oligotrophic, cold extreme lake ecosystems is limited. These organisms are likely to suffer prolonged physiological stress, which may account for the highly variable growth rates seen within and between Antarctic lakes. Accepted: 7 December 1999     

Response of methanotrophs and methane oxidation on ammonium application in landfill soils

To test the dose effect of ammonium (NH₄ ⁺) fertilization on soil methane (CH₄) oxidation by methanotrophic communities, batch incubations were conducted at a wide scale of NH₄ ⁺ amendments: 0, 100, 250, 500, and 1,000 mg N kg_dry soil ⁻¹. Denaturing gradient gel electrophoresis and real-time quantitative PCR analysis were conducted to investigate the correlation between the CH₄ oxidation capacity and methanotrophic communities. Immediately after the addition of NH₄ ⁺, temporal inhibition of CH₄ oxidation occurred, and this might have been due to the non-specific salt effect (osmotic stress). After a lag phase, the CH₄ oxidation rates of the soils with NH₄ ⁺ fertilization were promoted to levels higher than those of the controls. More than 100 mg N kg_dry soil ⁻¹ of NH₄ ⁺ addition resulted in the reduction of type II/type I MOB ratios and an obvious evolution of type II MOB communities, while less than 100 mg N kg_dry soil ⁻¹ of NH₄ ⁺ addition induced nearly no change of methanotrophic community compositions. The NH₄ ⁺-derived stimulation after the lag phase was attributed to the improvement of N availability for type I MOB. Compared with the controls, 100 mg N kg_dry soil ⁻¹ of NH₄ ⁺ addition doubled the CH₄ oxidation peak value to more than 20 mg CH₄ kg_dry soil ⁻¹ h⁻¹. Therefore, an appropriate amount of leachate irrigation on the landfill cover layer might efficiently mitigate the CH₄ emissions.