Cardiorespiratory responses to hypoxia in the African catfish, <Emphasis Type="Italic">Clarias gariepinus</Emphasis> (Burchell 1822), an air-breathing fish

The African catfish, Clarias gariepinus, possesses a pair of suprabranchial chambers located in the dorsal-posterior part of the branchial cavity having extensions from the upper parts of the second and fourth gill arches, forming the arborescent organs. This structure is an air-breathing organ (ABO) and allows aerial breathing (AB). We evaluated its cardiorespiratory responses to aquatic hypoxia. To determine the mode of air-breathing (obligate or accessory), fish had the respiratory frequency (f _R) monitored and were subjected to normoxic water (PwO₂ = 140 mmHg) without becoming hyperactive for 30 h. During this period, all fish survived without displaying evidences of hyperactivity and maintained unchanged f _R, confirming that this species is a facultative air-breather. Its aquatic O₂ uptake ( [(V)\dot]\textO₂ \dot{V}{\text{O}}_{2} ) was maintained constant down to a critical PO₂ (PcO₂) of 60 mmHg, below which [(V)\dot]\textO₂ \dot{V}{\text{O}}_{2} declined linearly with further reductions of inspired O₂ tension (PiO₂). Just above the PcO₂ the ventilatory tidal volume (V _T) increased significantly along with gill ventilation ( [(V)\dot]_\textG \dot{V}_{\text{G}} ), while f _R changed little. Consequently, the water convection requirement ( [(V)\dot]_\textG /[(V)\dot]\textO₂ ) \left( {\dot{V}_{\text{G}} /\dot{V}{\text{O}}_{2} } \right) increased steeply. This threshold applied to a cardiac response that included reflex bradycardia. AB was initiated at PiO₂ = 140 mmHg (normoxia) and air-breathing episodes increased linearly with more severe hypoxia, being significantly higher at PiO₂ tensions below the PcO₂. Air-breathing episodes were accompanied by bradycardia pre air-breath, to tachycardia post air-breath.     

Environment and feeding change the ability of heart rate to predict metabolism in resting Steller sea lions (<Emphasis Type="Italic">Eumetopias jubatus</Emphasis>)

The ability to use heart rate (fh) to predict oxygen consumption rates ( [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} ) in Steller sea lions and other pinnipeds has been investigated in fasting animals. However, it is unknown whether established fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationships hold under more complex physiological situations, such as when animals are feeding or digesting. We assessed whether fh could accurately predict [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} in trained Steller sea lions while fasting and after being fed. Using linear mixed-effects models, we derived unique equations to describe the fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationship for fasted sea lions resting on land and in water. Feeding did not significantly change the fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationship on land. However, Steller sea lions in water displayed a different fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} relationship after consuming a 4-kg meal compared with the fasting condition. Incorporating comparable published fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} data from Steller sea lions showed a distinct effect of feeding after a 6-kg meal. Ultimately, our study illustrated that both feeding and physical environment are statistically relevant when deriving [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} from telemetered fh, but that only environment affects the practical ability to predict metabolism from fh. Updating current bioenergetic models with data gathered using these predictive fh: [(V)\dot]_\textO2 \dot{V}_{{{\text{O}}_{2} }} equations will yield more accurate estimates of metabolic rates of free-ranging Steller sea lions under a variety of physiological, behavioral, and environmental states.     

Phenol biodegradation by the thermoacidophilic archaeon <Emphasis Type="Italic">Sulfolobus solfataricus</Emphasis> 98/2 in a fed-batch bioreactor

Toxic at low concentrations, phenol is one of the most common organic pollutants in air and water. In this work, phenol biodegradation was studied in extreme conditions (80°C, pH = 3.2) in a 2.7 l bioreactor with the thermoacidophilic archaeon Sulfolobus solfataricus 98/2. The strain was first acclimatized to phenol on a mixture of glucose (2000 mg l⁻¹) and phenol (94 mg l⁻¹) at a constant dissolved oxygen concentration of 1.5 mg l⁻¹. After a short lag-phase, only glucose was consumed. Phenol degradation then began while glucose was still present in the reactor. When glucose was exhausted, phenol was used for respiration and then for biomass build-up. After several batch runs (phenol < 365 mg l⁻¹), specific growth rate (μ_X) was 0.034 ± 0.001 h⁻¹, specific phenol degradation rate (q_P) was 57.5 ± 2 mg g⁻¹ h⁻¹, biomass yield (Y_X/P) was 52.2 ± 1.1 g mol⁻¹, and oxygen yield factor ( \textY_{\textX/\textO 2} ) \left( {{\text{Y}}_{{{\text{X}}/{\text{O}}_{ 2} }} } \right) was 9.2 ± 0.2 g mol⁻¹. A carbon recovery close to 100% suggested that phenol was exclusively transformed into biomass (35%) and CO₂ (65%). Molar phenol oxidation constant ( \textY_{\textO 2 /\textP} ) \left( {{\text{Y}}_{{{\text{O}}_{ 2} /{\text{P}}}} } \right) was calculated from stoichiometry of phenol oxidation and introducing experimental biomass and CO₂ conversion yields on phenol, leading to values varying between 4.78 and 5.22 mol mol⁻¹. Respiratory quotient was about 0.84 mol mol⁻¹, very close to theoretical value (0.87 mol mol⁻¹). Carbon dioxide production, oxygen demand and redox potential, monitored on-line, were good indicators of growth, substrate consumption and exhaustion, and can therefore be usefully employed for industrial phenol bioremediation in extreme environments.     

AMP-activated protein kinase plays a role in initiating metabolic rate suppression in goldfish hepatocytes

In the present study, we test the hypothesis that AMP-activated protein kinase (AMPK) initiates metabolic rate suppression in isolated goldfish hepatocytes. To accomplish this, we attempted to pharmacologically activate AMPK in goldfish hepatocytes with 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) and the thienopyridone, A769662, to examine the effects of AMPK activation on eukaryotic elongation factor-2 (eEF2), protein synthesis, and cellular oxygen consumption rate ( [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} ). Goldfish hepatocytes treated with 1 mM AICAR under normoxic conditions (>200 μM O₂) showed a modest but significant 1.1-fold increase in AMPK phosphorylation, a 7.5-fold increase in AMPK activity, a 1.4-fold increase in eEF2 phosphorylation, and a 24% decrease in [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} . At physiologically relevant [O₂] (<40 μM O₂), the addition of 1 mM AICAR resulted in only a 13% decrease in cellular [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} with no change in sensitivity to [O₂] as assessed by estimates of cellular P₅₀ and P₉₀ values. The addition of compound C, a general protein kinase inhibitor, after AICAR incubation did not reverse the effects of AICAR on [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} in normoxia. Treatment of hepatocytes with ≤200 μM A769662 did not affect AMPK activity, AMPK phosphorylation, eEF2 phosphorylation, or cellular [(M)\dot]_{\textO 2} \dot{M}_{{{\text{O}}_{ 2} }} . These data suggest that A769662 is not an activator of AMPK in goldfish hepatocytes. Although our study provides support for the hypothesis that AMPK plays a role in initiating metabolic rate suppression in goldfish hepatocytes, this support must be viewed cautiously because of the known off-target effects of the pharmacological agents used.     

Finite element–based injury metrics for pulmonary contusion via concurrent model optimization

This study explores the relationship between impact severity and resulting pulmonary contusion (PC) for four impact conditions using a rat model of the injury. The force–deflection response from a Finite Element (FE) model of the lung was simultaneously matched to experimental data from distinct impacts via a genetic algorithm optimization. Sprague-Dawley rats underwent right-side thoracotomy prior to impact. Insults were applied directly to the lung via an instrumented piston. Five cohorts were tested: a sham group and four groups experiencing lung insults of varying degrees of severity. The values for impact velocity (V) and penetration depth (D) of the cohorts were Group 1, (V = 6.0 m · s⁻¹, D = 5.0 mm), Group 2, (V = 1.5 m · s⁻¹,D = 5.0 mm), Group 3, (V = 6 m · s⁻¹, D = 2.0 mm), and Group 4, (V = 1.5 m · s⁻¹, D = 2.0 mm). CT scans were acquired at 24 h, 48 h, and 1 week post-insult. Contusion volume was determined through segmentation. FE-based injury metrics for PC were determined at 24 h and 1 week post-impact, based on the observed volume of contusion and first principal strain. At 24 h post-impact, the volume of high radiopacity lung (HRL) was greatest for the severe impact group (mean HRL = 9.21 ± 4.89) and was significantly greater than all other cohorts but Group 3. The concurrent optimization matched simulated and observed impact energy within one standard deviation for Group 1 (energy = 3.88 ± 0.883 mJ, observed vs. 4.47 mJ, simulated) and Group 2 (energy = 1.46 ± 0.403 mJ, observed vs. 1.50 mJ, simulated) impacts. Statistically significant relationships between HRL and impact energy are presented. The FEA-based injury metrics at 24 h post-contusion are e_max·[(e)\dot]_max{\varepsilon_{\max}\cdot \dot {\varepsilon}_{\max}} exceeding 94.5 s⁻¹, ε _max exceeding 0.284 and [(e)\dot]_max{\dot{\varepsilon}_{\max}} exceeding 470 s⁻¹. Thresholds for injury to the lung still present at 1 week post-impact were also determined. They are e_max·[(e)\dot]_max{\varepsilon_{\max}\cdot \dot {\varepsilon}_{\max}} exceeding 149 s⁻¹, ε _max exceeding 0.343 and [(e)\dot]_max{\dot{\varepsilon}_{\max}} exceeding 573 s⁻¹. A mesh sensitivity study found that thresholds based on strain rate were more sensitive to changes to mesh density than the threshold based on strain only.     

CO<Subscript>2</Subscript>-enriched microenvironment affects sucrose and macronutrients absorption and promotes autotrophy in the <Emphasis Type="Italic">in vitro</Emphasis> culture of kiwi (<Emphasis Type="Italic">Actinidia deliciosa</Emphasis> Chev. Liang and Ferguson)

In traditional in vitro culture, the low CO₂ concentration inside the vessels restricts photosynthesis and necessitates the addition of sucrose to the culture medium as the main energy source, thus bringing about changes in the absorption of mineral elements from the culture medium. In this study, we investigated macronutrient absorption and sugar consumption in Actinidia deliciosa Chevalier Liang and Ferguson cv. Hayward (kiwi), cultured on medium supplemented with varying amounts of sucrose (0, 10, and 20 g l⁻¹) under both heterotrophy and autotrophy, flushed with different concentrations of CO₂ (non-ventilation, 300, 600, and 2,000 μl l⁻¹). In ventilated systems with 20 g l⁻¹ of sucrose, sucrose absorption was less than under non-ventilation. The lowest rate of sucrose absorption was recorded when the explants were cultured on medium supplemented with 20 g l⁻¹ of sucrose and flushed with 600 μl l⁻¹ CO₂. Absorption of NO₃ ⁻, PO₄ ³⁻, and Mg²⁺ were high (maximum) at the end of the culture period (40 d) in explants flushed with 600 μl l⁻¹ CO₂ that have been cultured 20 d in the presence of sucrose and then transferred to a sucrose-free medium. These autotrophic conditions promoted maximum plant growth in terms of both fresh and dry mass as well as the length and number of shoots and leaves. The study shows that to maintain an optimum regime of mineral nutrition for prolonged culture of kiwi in vitro, an increased amount of these three ions should be supplemented in Murashige and Skoog’s medium.     

Role of the left aortic arch and blood flows in embryonic American alligator (<Emphasis Type="Italic">Alligator mississippiensis</Emphasis>)

All embryonic and fetal amniotes possess a ductus(i) arteriosus(i) that allows blood to bypass the pulmonary circulation and the non-functional lungs. The central hemodynamic of embryonic reptiles are unique, given the additional systemic aorta that allows pulmonary circulatory bypass, the left aorta (LAo). The LAo exits in the right ventricle or ‘pulmonary side’ of reptilian hearts in both embryos and adults, but its functional significance in ovo is unknown. This study investigated the role of the LAo in embryonic American alligators by surgically occluding the LAo and measuring oxygen consumption and, in addition, measured hemodynamic responses to hypoxia in embryonic alligators. We measured systemic cardiac output and primary chorioallantoic membrane (CAM) artery blood flow for normoxic and hypoxic-incubated (10% O₂) American alligator embryos (Alligator mississippiensis). Chronic blood flow (1–124 h) in the primary CAM artery for hypoxic-incubated embryos (92 ± 26 ml min⁻¹ kg⁻¹) was elevated when compared with normoxic-incubated embryos (29 ± 14 ml min⁻¹ kg⁻¹, N = 6; P = 0.039). For hypoxic-incubated embryos, acute LAo blood flow (49.6 ± 24.4 ml min⁻¹ kg⁻¹) was equivalent to the combined flow of the three systemic great vessels that arise from the left ventricle, the right aorta, common carotid and subclavian arteries (43.6 ± 21.5 ml min⁻¹ kg⁻¹, N = 5). Similarly, for normoxic-incubated embryos, LAo blood flow (27.3 ± 6.6 ml min⁻¹ kg⁻¹) did not statistically differ from the other three vessels (18.4 ± 4.9 ml min⁻¹ kg⁻¹, N = 5). This study contains the first direct test of LAo function and the first measurements of blood flow in an embryonic reptile. These data support the hypotheses that embryonic alligators utilize the LAo to divert a significant amount of right ventricular blood into the systemic circulation, and that CAM blood flow increases following chronic hypoxic conditions. However, surgical occlusion of the LAo did not affect egg [(V)\dot]_\textO2, \dot{V}_{{\text{O}}_{2}}, supporting the hypothesis that the LAo of reptiles is not critical to maintain in ovo oxygen consumption.     

Nocturnal lizards from a cool-temperate environment have high metabolic rates at low temperatures

Ectotherms from low-temperature environments have higher metabolic rates at low temperatures than those from warm-temperature environments. We predicted that nocturnal lizards, which are active at much lower environmental temperatures than diurnal lizards, would also have higher metabolic rates at low temperatures, and by association a lower thermal sensitivity (Q ₁₀) than diurnal and crepuscular lizards. We measured the rate of oxygen consumption ( [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} ) of eight cool-temperate species of lizard (four nocturnal, three diurnal, and one crepuscular) at 13 and 26°C and analyzed log transformations of these data using log mass as a covariate. As expected, [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} was positively correlated with temperature in all eight species, with [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} being two to four times higher at 26°C than at 13°C. As predicted, at 13°C (but not 26°C) the [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} was significantly higher in nocturnal than diurnal lizards. Species-specific differences and mass scaling factors explain the patterns of thermal sensitivity seen among these eight lizard species. Thermal sensitivity is strongly influenced by mass, with smaller species generally having higher thermal sensitivity of their metabolic rate, and this result deserves further exploration among other ectotherms. We conclude that, along with the previously reported lower cost of locomotion found in nocturnal lizards, they also partially offset the thermal handicap of activity at low body temperatures by having an elevated [(V)\dot]\textO ₂ \dot{V}{\text{O}}_{ 2} at lower temperatures.     

Oxygen consumption rate and Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase activity in early developmental stages of the sea urchin <Emphasis Type="Italic">Paracentrotus lividus</Emphasis> Lam.

Changes in oxygen consumption rate and Na⁺/K⁺-ATPase activity during early development were studied in the sea urchin Paracentrotus lividus Lam. The oxygen consumption rate increased from 0.12 μmol O₂ mg protein⁻¹ h⁻¹ in unfertilized eggs to 0.38 μmol O₂ mg protein⁻¹ h⁻¹ 25 min after fertilization. Specific activity of the Na⁺/K⁺-ATPase was significantly stimulated after fertilization, ranging up to 1.07 μmol P_i h⁻¹ mg protein⁻¹ in the late blastula stage and slightly lower values in the early and late pluteus stages.     

HNCA-TOCSY-CANH experiments with alternate <Superscript>13</Superscript>C-<Superscript>12</Superscript>C labeling: a set of 3D experiment with unique supra-sequential information for mainchain resonance assignment

Described here is a set of three-dimensional (3D) NMR experiments that rely on CACA-TOCSY magnetization transfer via the weak ³ \textJ_{\textCa\textCa} ^{ 3} {\text{J}}_{{{\text{C}}\alpha {\text{C}}\alpha }} coupling. These pulse sequences, which resemble recently described ¹³C detected CACA-TOCSY (Takeuchi et al. 2010) experiments, are recorded in ¹H₂O, and use ¹H excitation and detection. These experiments require alternate ¹³C-¹²C labeling together with perdeuteration, which allows utilizing the small ³ \textJ_{\textCa\textCa} ^{ 3} {\text{J}}_{{{\text{C}}\alpha {\text{C}}\alpha }} scalar coupling that is otherwise masked by the stronger ¹J_CC couplings in uniformly ¹³C labeled samples. These new experiments provide a unique assignment ladder-mark that yields bidirectional supra-sequential information and can readily straddle proline residues. Unlike the conventional HNCA experiment, which contains only sequential information to the ^{1 3} \textC^a ^{ 1 3} {\text{C}}^{\alpha } of the preceding residue, the 3D hnCA-TOCSY-caNH experiment can yield sequential correlations to alpha carbons in positions i−1, i + 1 and i−2. Furthermore, the 3D hNca-TOCSY-caNH and Hnca-TOCSY-caNH experiments, which share the same magnetization pathway but use a different chemical shift encoding, directly couple the ¹⁵N-¹H spin pair of residue i to adjacent amide protons and nitrogens at positions i−2, i−1, i + 1 and i + 2, respectively. These new experimental features make protein backbone assignments more robust by reducing the degeneracy problem associated with the conventional 3D NMR experiments.     

Average shear rate in three pneumatic bioreactors

A method proposed in recent literature was applied to evaluate the average shear rate ( [(g)\dot]_av ) \left( {\dot{\gamma }_{\rm av} } \right) in three pneumatic bioreactors of 5-dm³ working volume: bubble column, split airlift, and concentric-tube airlift. The volumetric oxygen transfer coefficient (k _L a) is the appropriate characteristic parameter to assess the average shear rate ( [(g)\dot]_av ) \left( {\dot{\gamma }_{\rm av} } \right) in this methodology. Correlations for [(g)\dot]_av \dot{\gamma }_{\rm av} as a function of superficial gas velocity in the riser region (U _GR) and rheological fluid properties (consistency index, K, and flow index, n) were obtained for each model of pneumatic bioreactor studied. The [(g)\dot]_av \dot{\gamma }_{\rm av} values estimated by the proposed methodology lay within the range of values calculated by classical correlations. The proposed correlations were utilized to predict the [(g)\dot]_av \dot{\gamma }_{\rm av} during the Streptomyces clavuligerus cultivations carried out at the same specific air flow rate (3.5 vvm) in the different types of pneumatic bioreactors. The lowest values of [(g)\dot]_av \dot{\gamma }_{\rm av} related to the highest values of consistency index (K) were found for the bubble column bioreactor, and the highest values of [(g)\dot]_av \dot{\gamma }_{\rm av} related to the lowest values of K were found for the concentric-tube airlift bioreactor. Intermediate values were found for the split airlift bioreactor. The results showed that high [(g)\dot]_av \dot{\gamma }_{\rm av} values affect the structural health of the mycelia by the rupture of the hipha.     

Biochemical characterization of the carotenoid 1,2-hydratases (CrtC) from <Emphasis Type="Italic">Rubrivivax gelatinosus</Emphasis> and <Emphasis Type="Italic">Thiocapsa roseopersicina</Emphasis>

Two carotenoid 1,2-hydratase (CrtC) genes from the photosynthetic bacteria Rubrivivax gelatinosus and Thiocapsa roseopersicina were cloned and expressed in Escherichia coli in an active form and purified by affinity chromatography. The biochemical properties of the recombinant enzymes and their substrate specificities were studied. The purified CrtCs catalyze cofactor independently the conversion of lycopene to 1-HO- and 1,1′-(HO)₂-lycopene. The optimal pH and temperature for hydratase activity was 8.0 and 30°C, respectively. The apparent K _m and V _max values obtained for the hydration of lycopene were 24 μM and 0.31 nmol h⁻¹ mg⁻¹ for RgCrtC and 9.5 μM and 0.15 nmol h⁻¹ mg⁻¹ for TrCrtC, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed two protein bands of 44 and 38 kDa for TrCrtC, which indicate protein processing. Both hydratases are also able to convert the unnatural substrate geranylgeraniol (C20 substrate), which functionally resembles the natural substrate lycopene.     

Plasma free amino acid kinetics in rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) using a bolus injection of <Superscript>15</Superscript>N-labeled amino acids

To gain insight into the metabolic design of the amino acid carrier systems in fish, we injected a bolus of ¹⁵N amino acids into the dorsal aorta in mature rainbow trout (Oncorhynchus mykiss). The plasma kinetic parameters including concentration, pool size, rate of disappearance (R _d), half-life and turnover rate were determined for 15 amino acids. When corrected for metabolic rate, the R _d values obtained for trout for most amino acids were largely comparable to human values, with the exception of glutamine (which was lower) and threonine (which was higher). R _d values ranged from 0.9 μmol 100 g⁻¹ h⁻¹ (lysine) to 22.1 μmol 100 g⁻¹ h⁻¹ (threonine) with most values falling between 2 and 6 μmol 100 g⁻¹ h⁻¹. There was a significant correlation between R _d and the molar proportion of amino acids in rainbow trout whole body protein hydrolysate. Other kinetic parameters did not correlate significantly with whole body amino acid composition. This indicates that an important design feature of the plasma-free amino acids system involves proportional delivery of amino acids to tissues for protein synthesis.     

Temporal variation in CO<Subscript>2</Subscript> efflux from soil and snow surfaces in a Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) plantation,central Japan

CO₂ efflux from soil and snow surfaces was measured continuously in a Japanese cedar (Cryptomeria japonica D. Don) forest in central Japan using an open dynamic chamber system. The chamber opens and closes automatically and records measurements based on an open-flow dynamic method. Between May and December, mean soil CO₂ efflux ranged from 1,529 mg CO₂ m⁻² h⁻¹ in September to 255 mg CO₂ m⁻² h⁻¹ in December. The seasonal change in CO₂ efflux from the soil paralleled the seasonal pattern of soil temperature. No marked diurnal trends in soil CO₂ efflux were observed on days without rainfall, whereas significant pulses in soil CO₂ efflux were observed on days with rainfall. In this plantation, soil CO₂ efflux frequently responded to rainfall. Measurements of changes from litter-covered soil to snow-covered surfaces revealed that CO₂ efflux decreased from values of ca. 250 mg CO₂ m⁻² h⁻¹ above soil to less than 33 mg CO₂ m⁻² h⁻¹ above snow. Soil temperature alone explained 66% of the overall variation in soil CO₂ efflux, but explained approximately 85% of the variation when data from two anomalous periods were excluded. Moreover, we found a significant correlation between soil CO₂ efflux and soil moisture (which explained 44% of the overall variation) using a second-order polynomial function. Our results suggest that the seasonality of CO₂ efflux is affected not only by soil temperature and moisture, but also by drying and rewetting cycles and by litterfall pulses.     

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.     

Quasi steady state growth of <Emphasis Type="Italic">Lactococcus lactis</Emphasis> in glucose-limited acceleration stat (A-stat) cultures

Quasi steady state growth of Lactococcus lactis IL 1403 was studied in glucose-limited A-stat cultivation experiments with acceleration rates (a) from 0.003 to 0.06 h⁻² after initial stabilization of the cultures in chemostat at D = 0.2–0.3 h⁻¹. It was shown that the high limit of quasi steady state growth rate depended on the acceleration rate used—at an acceleration rate 0.003 h⁻² the quasi steady state growth was observed until μ _crit = 0.59 h⁻¹, which is also the μ _max value for the culture. Lower values of μ _crit were observed at higher acceleration rates. The steady state growth of bacteria stabilized at dilution rate 0.2 h⁻¹ was immediately disrupted after initiating acceleration at the highest acceleration rate studied—0.06 h⁻². Observation was made that differences [Δ(μ − D)] of the specific growth rates from pre-programmed dilution rates were the lowest using an acceleration rate of 0.003 h⁻² (< 4% of preset changing growth rate). The adaptability of cells to follow preprogrammed growth rate was found to decrease with increasing dilution rate—it was shown that lower acceleration rates should be applied at higher growth rates to maintain the culture in the quasi steady state. The critical specific growth rate and the biomass yields based on glucose consumption were higher if the medium contained S ₀ = 5 g L⁻¹ glucose instead of S ₀ = 10 g L⁻¹. It was assumed that this was due to the inhibitory effect of lactate accumulating at higher concentrations in the latter cultures. Parallel A-stat experiments at the same acceleration and dilution rates showed good reproducibility—Δ(μ − D) was less than 5%, standard deviations of biomass yields per ATP produced (Y _ATP), and biomass yields per glucose consumed (Y _XS) were less than 15%.     

Effect of 24-Epibrassinolide on Chlorophyll Fluorescence and Photosynthetic CO<Subscript>2</Subscript> Assimilation in <Emphasis Type="Italic">Vicia faba</Emphasis> Plants Treated with the Photosynthesis-Inhibiting Herbicide Terbutryn

Simultaneous measurements of chlorophyll (Chl) fluorescence and CO₂ assimilation (A) in Vicia faba leaves were taken during the first weeks of growth to evaluate the protective effect of 24-epibrassinolide (EBR) against damage caused by the application of the herbicide terbutryn (Terb) at pre-emergence. V. faba seeds were incubated for 24 h in EBR solutions (2 × 10⁻⁶ or 2 × 10⁻⁵ mM) and immediately sown. Terb was applied at recommended doses (1.47 or 1.96 kg ha⁻¹) at pre-emergence. The highest dose of Terb strongly decreased CO₂ assimilation, the maximum quantum yield of PSII photochemistry in the dark-adapted state (F _V/F _M), the nonphotochemical quenching (NPQ), and the effective quantum yield (ΔF/F′_M) during the first 3–4 weeks after plant emergence. Moreover, Terb increased the basal quantum yield of nonphotochemical processes (F ₀/F _M)_, the degree of reaction center closure (1 − q _p), and the fraction of light absorbed in PSII antennae that was dissipated via thermal energy dissipation in the antennae (1 − F′_V/F′_M). The herbicide also significantly reduced plant growth at the end of the experiment as well as plant length, dry weight, and number of leaves. The application of EBR to V. faba seeds before sowing strongly diminished the effect of Terb on fluorescence parameters and CO₂ assimilation, which recovered 13 days after plant emergence and showed values similar to those of control plants. The protective effect of EBR on CO₂ assimilation was detected at a photosynthetic photon flux density (PFD) of 650 μmol m⁻² s⁻¹ and the effect on ΔF/F′_M and photosynthetic electron transport (J) was detected under actinic lightings up to 1750 μmol m⁻² s⁻¹. The highest dose of EBR also counteracted the decrease in plant growth caused by Terb, and plants registered the same growth values as controls.     

Soil temperature and biotic factors drive the seasonal variation of soil respiration in a maize (Zea mays L.) agricultural ecosystem

The diurnal and seasonal variation of soil respiration (SR) and their driving environmental factors were studied in a maize ecosystem during the growing season 2005. The diurnal variation of SR showed asymmetric patterns, with the minimum occurring around early morning and the maximum around 13:00 h. SR fluctuated greatly during the growing season. The mean SR rate was 3.16 μmol CO₂ m⁻² s⁻¹, with a maximum of 4.87 μmol CO₂ m⁻² s⁻¹ on July 28 and a minimum of 1.32 μmol CO₂ m⁻² s⁻¹ on May 4. During the diurnal variation of SR, there was a significant exponential relationship between SR and soil temperature (T) at 10 cm depth: . At a seasonal scale, the coefficient α and β fluctuated because the biomass (B) increased α, and the net primary productivity (NPP) of maize markedly increased β of the exponential equation. Based on this, we developed the equation to estimate the magnitude of SR and to simulate its temporal variation during the growth season of maize. Most of the temporal variability (93%) in SR could be explained by the variations in soil temperature, biomass and NPP of maize. This model clearly demonstrated that soil temperature, biomass and NPP of maize combined to drive the seasonal variation of SR during the growing season. However, only taking into account the influence of soil temperature on SR, an exponential equation over- or underestimated the magnitude of SR and resulted in an erroneous representation of the seasonal variation in SR. Our results highlighted the importance of biotic factors for the estimation of SR during the growing season. It is suggested that the models of SR on agricultural sites should not only take into account the influence of soil temperature, but also incorporate biotic factors as they affect SR during the growing season.     

Nitric oxide promotes in vitro organogenesis in <Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.

The effects of nitric oxide (NO) on caulogenesis, shoot organogenesis and rhizogenesis from hypocotyl explants of Linum usitatissimum were investigated. Exogenously supplied NO donors, 5 μM sodium nitroprusside (SNP), 2 μM S-nitroso-N-acetylpenicillamine (SNAP) and 2 μM 3-morpholinosydnonimine (SIN-1), significantly promoted shoot differentiation from the hypocotyl explants of L. usitatissimum excised from its in vitro raised seedlings. Potassium ferrocyanide, a structural analogue of SNP, lacking NO group, did not promote shoot organogenesis. Likewise, products of NO, \textNO₂ ^- {\text{NO}}_{2}^{ - } and \textNO₃ ^- {\text{NO}}_{3}^{ - } supplied as 5 μM NaNO₂ and 5 μM NaNO₃ did not enhance shoot differentiation. Another source of NO, a mixture of sodium nitrite (SN) provided along with ascorbic acid (AsA), also caused significant promotion in the average number of shoots per responding explant. SNP also augmented the rhizogenic response of the microshoots in terms of percentage of responding explants, number of roots per responding explant and average root length. The NO scavengers, 2-(4-carboxy-phenyl)-4, 4, 5, 5-tetramethylimideazoline-1-oxyl-3-oxide (cPTIO) or methylene blue (MB), provided along with SNP, SNAP, SIN-1 or SN + AsA, at concentrations equimolar to the optimum concentration of the donors, reversed the promotory influence, thereby, confirming the role of NO in promotion of in vitro morphogenesis. However, NO scavengers individually did not affect the observed morphogenic processes. Morphological and histological studies of hypocotyl segments cultured on BM or BM + SNP for 4, 8 and 12 days demonstrated that SNP enhanced shoot differentiation by inducing a higher number of shoot primordia, each of which develops into a single shoot.