Growth and product formation of actinomycetes cultivated at increased total pressure and oxygen partial pressure

Summary Influence of pressure (P) and oxygen partial pressure ( ) on cultivation of various Streptomyces spp. and Micromonospora purpurea was examined in pressurized air-lift and stirred tank fermenters. The maximum was 2100 mbar. Growth and product formation of all cultures tested were markedly influenced by higher than 1000 mbar. There is evidence that wild strains are more oxygen tolerant than production strains. At a certain the metabolic activities of all cultures were inhibited. However, results obtained with S. aureofaciens and S. rimosus indicated an increase in specific product formation rate at elevated pressure. With increase in oxygen tension incorporation of oxygen into tetracycline molecules was enhanced. Since elevated oxygen tension can either show inhibiting effects or may be used for regulation of product formation and selectivity, the influence of should be determined in an appropriate experimental set-up for each process.Offprint requests to: U. Onken     

Control of acid-base status in active and dormant land snails,Otala lactea (Pulmonata,Helicidae)

Summary Control of extracellular acid-base status was examined during activity and dormancy inOtala lactea (Pulmonata, Helicidae). Active snails showed little variation in hemolymph pH and at constant temperature. With increase of temperature, hemolymph increased from about 6 Torr at 5°C to 13 Torr at 24°C and pH decreased by about 0.017 pH units/°C, a pattern consistent with alphastat regulation of pH via ventilatory control of .During dormancy, mean hemolymph increased to about 50 Torr. Venous pH declined by about 0.4 units due to hypercapnia and fluctuated more widely than in active snails due to variability of . Hemolymph pH declined further in prolonged dormancy due to progressive metabolic acidosis; after one year of dormancy the mean hemolymph pH was about 0.8 units lower than that of active snails at similar temperature.Active snails exposed experimentally to high showed a large increase in hemolymph [HCO ₃ ^– ]. However, [HCO ₃ ^– ] declined by up to 50% during dormancy, despite the naturally occurring hypercapnia. Hemolymph osmolality and the concentrations of solutes other than [HCO ₃ ^– ] increased with increasing duration of dormancy. Concentrations of magnesium and calcium increased about 2.5 times more rapidly than those of sodium and chloride, indicating that acidosis is partially offset by the dissolution of carbonates from the shell or tissues.     

Reversed bohr and root shifts in hemocyanin of the marine prosobranch,Buccinum undatum: Adaptations to a periodically hypoxic habitat

Summary Oxygen binding properties of the hemocyanin-containing blood ofBuccinum undatum were examined in vitro and in vivo under normoxic ( 150 mmHg) and hypoxic ( 50 mmHg) conditions at 10°C. Blood pH and showed a decrease in vivo under hypoxic conditions. Oxygen uptake at high water , was about 18 ml O₂/kg·h (wet weight) and the critical oxygen tension between 25 and 50 mm Hg. In vitro the O₂ binding to hemocyanin showedn-values independent of pH, while both O₂ affinity and oxygen carrying capacity were strongly pH dependent. Oxygen affinity increased below pH=8.1 and thus showed a pronounced reversed Bohr shift in the physiological pH range (7.5<pH<8.1). The oxygen carrying capacity similarly increased markedly with falling pH in the physiological pH range (reversed Root shift). Astrup titration curves showed a metabolic and respiratory acidosis under hypoxic conditions ( 50 mm Hg). The role of hemocyanin in the transport of oxygen in relation to ambient O₂ availability is discussed.     

Estimation of surface charges in some biological membranes

Summary The resting membrane potential data existing in the literature for the giant axon of the squid, frog muscle and barnacle muscle have been analyzed from the standpoint of the theory of membrane potential due to Kobatake and co-workers. The average values derived for the effective charge density (where is a constant, , and represents the fraction of counterions that are free, and is the stoichiometric charge density in the membrane) present on the different biomembranes existing in their normal ionic environment are 0.3, 0.325 and 0.17 M for the squid axon, frog and barnacle muscles, respectively. On the assumption that the values of are 0.4 and 0.2 for nerve and muscle membranes, respectively, values of 0.75, 1.62 and 0.85 M have been derived for the stoichiometric charge density present in the respective biological membranes. These correspond to 1 negative charge per 222, 103 and 195 Å of the membrane area of the squid axon, frog and barnacle muscles, respectively.     

Activity and oxygen consumption during hypoxic exposure in high altitude and lowland sceloporine lizards

Summary Resting rates of O₂ consumption against , exercise endurance times and during recovery from vigorous exercise were measured inSceloporus occidentalis captured near sea level and inS. graciosus captured above 2850 m. Oxygen consumption against was also measured inS. occidentalis captured above 2850 m. When was recorded continuously, as ambient was slowly reduced from 155 Torr, it became directly dependent upon ambient between 110 and 120 Torr. The critical for the high altitude lizards was lower than that for the lowland lizards, which enabled the former to maintain relatively higher 's when ambient was reduced below 120 Torr. The high altitude lizards also had significantly greater endurance when stimulated to exercise at 1600 m ( 130 Torr). Both the higher under hypoxia and the greater endurance roughly parallel a significantly greater maximum in the high altitude lizards. At a simulated altitude of 3600 m ( 100 Torr), maximum and rate of recovery of the O₂ debt calculated from post active were significantly reduced in the lowland but not the high altitude lizards. The effects of simulated altitude conditions on the lowland but not the mountaine animals indicate adaptations to altitude in these sceloporine lizards. We did not find any consistent relationship between organ/body weight ratios or hematocrit and our measures of endurance or the altitude at which the lizards were captured.     

Anaerobic metabolism in aquatic insect larvae: studies onChironomus thummi andCulex pipiens

Summary The metabolic effect of hypoxia and anoxia on the larvae ofChironomus thummi andCulex pipiens was investigated. InC. thummi anoxia resulted in a characteristic decrease of ATP and P-arginine concentrations and in an accumulation of alanine and lactate within 60 minutes. These changes continued during prolonged incubation but at lower rates. Ethanol, the major product during long-term anoxia, was largely excreted into the ambient water.A significant accumulation of these metabolites occurred only at a of 7 Torr. However, the proportion of anaerobic energy production even at this low amounted to less than 5% of the total energy consumption measured during experimental anoxia. Thus the chironomid larvae exhibited a remarkable capacity for utilizing very low levels of oxygen to maintain an aerobic metabolism. Complete anaerobiosis was observed only under anoxic conditions.Recovery from prior anoxia began with the reestablishment of normal ATP, P-arginine and succinate concentrations, whereas removal of the accumulated alanine and lactate and replenishment of the normally high level of malate required several hours. Culex larvae were shown to have a very low anaerobic capacity and a high rate of lactate accumulation.The significance of the results is discussed with particular emphasis on comparative aspects.     

Anaerobic energy metabolism of the European eel,Anguilla anguilla L.

Summary Eels, acclimated the 15°C and aerated water (P _{O 2} 130 mm Hg) were exposed to hypoxia (P _{O 2} lowered from 130 to 8 mm Hg in 4 h) and to complete anoxia until loss of equilibrium. Experiments were carried out at night. The mean survival time (LT₅₀) during anoxic conditions proved to be 5.7 h. ATP, ADP, AMP, IMP, CrP, glycogen, lactate, pyruvate, -ketoglutarate, malate, succinate, alanine, aspartate, glutamate and ammonia levels were determined in skeletal muscle and liver of control, hypoxic and anoxic fish. Some of the mentioned parameters were also measured in heart muscle and blood. Hypoxia causes declines of aspartate (muscle), CrP (muscle) and glycogen (liver, heart), and increases of alanine (blood, liver) and lactate (blood, liver, heart). During anoxia, muscle CrP stores are almost completely exhausted and adenylates are partially broken down to IMP. A decrease of glycogen and an accumulation of lactate were observed in all tissues examined. The energy charge of muscle and heart did not drop below 0.79, but in liver tissue it decreased from 0.65 to 0.17. Liver cytoplasm became significantly reduced during anoxia, but such a change of redox state did not occur in muscle. Eels seem to lack the capacity for anaerobic fermentation of glycogen to ethanol, as observed in goldfish. Lactate glycolysis and creatine phosphate breakdown appear to be the main energy producing pathways during anaerobiosis.Abbreviations ALA alanine - ASP aspartate - CrP creatine phosphate - EC (adenylate) energy charge - GLU glutamate - GLC glucose - GLY glycogen - IMP inosine-5-monophosphate - KG ketoglutarate - LAC lactate - MAL malate - PYR pyruvate - SUC succinate - TAN total pool of adenine nucleotides     

Oxygen controlled batch cultivations of Schizophyllum commune for enhanced production of branched β-1,3-glucans

Oxygen and shear stress are the key factors for enhanced glucan production with Schizophyllum commune. During batch cultivation control of or (specific oxygen uptake rate) was achieved by variation of the impeller speed. Biomass was modelled by using the carbon and oxygen balance derived from exhaust data. At mycel growth a of 0.042 h^–1 presents just the border before oxygen limitation arises and is simultaneously the optimum operation condition for maximum glucan formation. Related to an overall cultivation time of 72 h a maximum of both productivity (4.3 kg m^–3 d^–1) and yield (13 kg m^–3) were obtained.List of Symbols C kg m^–3 concentration - k _L a h ^–1 volume related oxygen transfer coefficient - K _s mol m^–3 substrate saturation constant - N rpm impeller speed - % oxygen partial pressure of the liquid phase - kg m^–3h^–1 oxygen uptake rate - h^–1 specific oxygen uptake rate, kg O₂ (kg biomass h)^–1 - t h time - yield coefficient (biomass formed/oxygen consumed) Greek Symbols h^–1 specific growth rate Indices O ₂ oxygen - X biomass - L liquid phase - * gas/liquid interface - S substrate (glucose) Dedicated to the 65th birthday of Professor Fritz Wagner.This work was kindly supported in parts by B. Braun Biotech International. The authors are grateful to Prof. Dr. Fritz Wagner for scientific support and appreciate the technical assistance of Detlev Rasch     

Water economy in bird nests

Summary Nest humidity ( ) under an incubating bantam hen was measured at ambient ranging from 1.3 to 25.0 mmHg. Weight loss of eggs was recorded in clutches of varying size. Nest and ambient were also measured in wild incubating willow ptarmigan nests in dry and wet habitats.Nest increased linearly with ambient in a way predictable on the assumption that the water vapour conductance ( ) of brood patch skin, plumage and eggs were constant and independent of ambient . Nest was also dependent of clutch size. Egg dehydration was quantitatively predicted from measured values and the laws of diffusion.Our findings confirm earlier conclusions that the adult bird does not actively regulate nest at varying ambient . Birds can presumably achieve appropriate egg dehydration by a strategy combining nest site, nest construction, egg shell conductance and incubation behaviour which meets the requirements of their breeding climate.Abbreviations water vapur pressure - water vapur conductance - water flux     

Gas exchange in the incubation mounds of megapode birds

Summary The brush turkey (Alectura lathami) and mallee fowl (Leipoa ocellata) are megapode birds that incubate their eggs by burying them in mounds. Respiratory gas exchange between the buried eggs and the atmosphere occurs mainly by diffusion through about 60 cm of decomposing forest litter (brush turkey) or sand (mallee fowl).Gas fluxes in the brush turkey mound are greatly influenced by the respiration of thermophilic microorganisms which consume O₂ at rates over eight times that of all of the eggs. The respiratory exchange ratio ( ) of the microorganisms is 0.75 and theQ ₁₀ for metabolism is 2.56. Fermentation and nitrogen fixation do not occur in the mounds.If the mound becomes too wet, gas tensions near the eggs can become critical because water increases rates of microbial respiration and impedes gas diffusion. However, field mounds are relatively dry, possibly because the adult bird modifies the shape of the mound and affects the entry of rain water. At egg level in field mounds, and are about 132 and 21 Torr, respectively, in both species. Embryonic respiration decreases and increases about 5 Torr in the immediate environment of individual eggs in late development. Due to a high eggshell gas conductance, which increases during incubation, the gas tensions within the shell of late embryos ( ca. 108 Torr, ca. 47 Torr) are not far from the mean values found in species that nest above ground.     

Gill function in an elasmobranch

Summary Highly efficient oxygen uptake in elasmobranchs, as indicated by frequent excess of over has previously been ascribed to the operation of multicapillary rather than counter-current gas exchange by the gills. Analysis of models shows that, at maximum efficiency, a multicapillary system cannot account for values of greater than . In Port Jackson sharks Heterodontus portusjacksoni) commonly exceeds , which indicates the operation of a functional counter-current at the respiratory surface. The anatomical basis of this counter-current is provided by the demonstration that a continuous flow of water passes between the secondary lamellae into septal canals and thence via the parabranchial cavities to the exterior.Queen Elizabeth II Fellow.     

Respiratory responses to long-term temperature exposure in the box turtle,Terrapene ornata

Summary In late February, seven box turtles were collected with body temperatures between 7 and 9°C. Ventilation, gas exchange and end-tidal and were recorded at 5, 10, 15 and 25°C, the animals being at each temperature for 2 to 3 weeks. There was a pronounced diurnal rhythm of breathing frequency at all temperatures. At 5°C the mean 24-h frequency was only 3.7 breaths h^–1. At 15°C the frequency was 16 times higher with a 17-fold increase of ventilation. Oxygen uptake only changed from 3.4 to 12.7 ml·kg^–1·h^–1. Consequently, the ratio (ventilation, ml BTPS/O₂ uptake, ml STPD) increased from 12.5 at 5°C to 48 at 15°C, but decreased to 24 at 25°C. The decrease of this ratio during cold exposure contrasts with an increase of the ratio during cooling earlier reported for fresh water turtles,Pseudemys. Cutaneous CO₂ elimination was important at low temperature. This caused a decrease of the pulmonary gas exchange ratio so that end-tidal remained low at 5°C in spite of an end-tidal of only 54 Torr.     

Altitudinal and seasonal effects on aerobic metabolism of deer mice

Summary I compared the maximal aerobic metabolic rates ( ), field metabolic rates (FMR), aerobic reserves ( -FMR), and basal metabolic rates (BMR) of wild and recently captured deer mice from low (440 m) and high (3800 m) altitudes. To separate the effects of the thermal environment from other altitudinal effects, I examined mice from different altitudes, but similar thermal environments (i.e., summer mice from high altitude and winter mice from low altitude). When the thermal environment was similar, , FMR, and aerobic reserve of low and high altitude mice did not differ, but BMR was significantly higher at high altitude. Thus, in the absence of thermal differences, altitude had only minor effects on the aerobic metabolism of wild or recently captured deer mice.At low altitude, there was significant seasonal variation in , FMR, and aerobic reserve, but not BMR. BMR was correlated with , but not with FMR. The significant positive correlation of BMR with indicates a cost of high , because higher BMR increases food requirements and energy use during periods of thermoneutral conditions.Abbreviations BMR basal metabolic rate - FMR field metabolic rate - partial pressure of oxygen - T _a ambient temperature - T _b body temperature - T _e operative temperature - maximal aerobic metabolic rate     

Influences of exercise-stress and adrenaline upon intra- and extracellular acid-base status,electrolyte composition and respiratory properties of blood in tench (Tinca tinca) at different seasons

Summary Exercise-stress in tench resulted in severe acidoses in both the red cells and the extracellular fluid in vivo. These coincident pH decreases conformed to the in vitro pH_i-pH_e relationship for tench blood in the oxygenated state. The extracellular acidosis was primarily respiratory in winter and metabolic in spring and summer. This was due to more effective buffering of metabolic protons in winter by an elevation in [HCO ₃ ^– ] levels, rather than to differences in the lactic- and carbonic acid loads. A good correspondence was found between buffered metabolic protons and increases in [lactate].There was no evidence for -adrenergic red cell swelling and associated red cell pH changes in tench both after exercise and adrenaline infusion. Arterial O₂ transport was, however, improved in exercise by pronounced increases in .Large increases in plasma potassium concentration and small elevations of chloride and calcium levels occurred in exercise. Hematocrit and blood [Hb] also increased, probably due to an adrenergic release of erythrocytes from the spleen, but these increases were small and appeared unimportant for blood O₂ transport.Seasonal differences were found in exercise-induced changes in [lactate], in the magnitude of electrolyte and changes, as well as in resting values for pH_e, pH_i, [HCO ₃ ^– ], [Cl^–] and [Ca⁺⁺]. The origin and importance of these are discussed.     

Cardiovascular effects of parathyroid hormone in the frog,Rana catesbeiana

Summary Mean arterial blood pressure ( ), heart rate (HR), and cardiac isometric contractile force (CF) were recorded simultaneously in the anesthetized frog,Rana catesbeiana, and the effects of the synthetic 1–34 peptide of bovine parathyroid hormone, bPTH-(1-34), on these cardiovascular parameters were tested. Doses of 10, 20, 40, 80, and 100 U·kg^–1 of PTH were administered i.v. and the effects noted. Isotonic (frog) saline was used as control. Following bPTH-(1-34), decreased in dose-dependent fashion, reaching maximum within 60 s post-injection with a duration of 7–8 min. CF was shown for the first time to decrease in dose-dependent fashion, reaching a maximum within 70 s and exhibiting a duration of approximately 8 min. No effect of bPTH-(1-34) on heart rate was noted.Abbreviations bPTH-(1-34) 1-34 peptide of bovine parathyroid hormone - mean arterial pressure - HR heart rate - CF cardiac contractile force - U international units - SR sarcoplasmic reticulum     

Ventilation and gas exchange during rest and exercise in adult green sea turtles

Summary Ventilation and metabolic rate were measured during exercise in adult female green turtles at Tortuguero, Costa Rica. Six turtles were studied at night on the beach while actively covering their nests. Five turtles, captured after nesting, were studied at rest, during 20 min of spontaneous activity, and during recovery from the activity. Arterial blood samples were obtained from the latter animals and analyzed for pH, , O₂ concentration and lactate concentration. Blood was obtained by heart puncture from 8 turtles immediately after nesting and analyzed for blood lactate. Active metabolism ( ) in both groups was almost 10 times the standard resting value (0.024 l/kg·h). The increase in ventilation during exercise, due exclusively to higher breathing frequency, exceeded the increase in , so that the ratio (the air convection requirement), more than doubled. The respiratory exchange ratio, , that averaged 0.56 in the resting turtles, increased to 1.08 during exercise in the captured turtles and was 0.90 in the nesting animals. Arterial and O₂ saturation remained unchanged during exercise, indicating efficient gas exchange in the lungs. Pre-exercise values of all variables were restored 1 h after the end of exercise. Blood acid-base changes associated with activity in the captive turtles were variable and not statistically significant, but suggested partially compensated metabolic acidosis. Lactate concentrations were significantly elevated in the nesting turtles.     

Effect of chronic acidosis on discharge frequency of intrapulmonary chemoreceptors in the chicken

Summary In order to determine the pH effect on the discharge of intrapulmonary chemoreceptors (IPC) not mediated through , we determined discharge frequency-ln relationships of 28 IPC from ten adult cockerels made acidotic by ingestion of feed containing 6% NH₄Cl for 2 to 5 days. Blood buffer curves — the relationships between pH and — from these cockerels were shifted in parallel towards acidosis by 0.24 pH units (SE=±0.05) compared to control curves. The average slope of the IPC discharge frequenc-ln relationships was less negative than that reported for similar cockerels with normal acid-base balance and the average intercept was decreased. We conclude that pH has an effect on IPC discharge, independent of changes in ; this effect decreases IPC sensitivity to CO₂. We calculate that a shift towards acidosis in the blood buffer curve of one pH unit causes the slope of the IPC discharge frequency-ln relationship to be less negative by 14.7±3.3 imp (s·ln )^–1 and that for any acidotic shift in the blood buffer curve (pH), the average IPC discharge frequency=[–10.7+(14.7·pH)][ln( /25.5)]+3.37. Since IPC discharge is altered by changes in acid-base balance, IPC may contribute to the implementation of respiration responses during metabolic disturbances.Abbreviation IPC intrapulmonary chemoreceptor     

Carbonic anhydrase activity in the blood and the gills of rainbow trout during long-term hypercapnia in hard,bicarbonate-rich freshwater

Summary Carbonic anhydrase (CA) activities in gills and venous blood, acid-base balance, and haematological variables were studied during environmental hypercapnia in rainbow trout (Salmo gairdneri). Batches of 8–10 fish were exposed to about 3 or 13 mmHg in flow-through tests of various duration from 4 h to 80 days.After initial acidosis, blood pH rose above pre-experimental values. At 3 mmHg it became normal again within 21 days, while at 13 mmHg the overshoot lasted for 80 days. In fish acclimated for 3 weeks or more to 13 mmHg , blood HCO ₃ ^– increased four to five times while plasma Cl^– levels were lower and K⁺ higher. Na⁺ levels did not show any consistent trend associated with exposure to hypercapnia. After an initial acidaemia, Hct, Hb, and RBC remained relatively constant.Patterns of change in CA activity differed between gills and erythrocytes. Initially, blood CA decreased at both levels. It then began rising after about 3 weeks and tended to reach pre-experimental values by 80 day's hypercapnia. At 13 mmHg , gill CA increased to twice the pre-experimental level. Compared with blood CA, gill CA appeared to be more specifically involved in fish acclimation to hypercapnia, which demands an increase in blood bicarbonate to provide a sufficient buffering capacity. Increased CA indicates that the gill enzyme may play a more important role than blood CA in acid-base regulation in fish during hypercapnia.Abbreviations CA carbonic anhydrase - Hb haemoglobin - Hct haematocrit value - RBC red blood cells     

Effects of temperature and altitude on ventilation and gas exchange in chukars (Alectoris chukar)

Summary The effects of ambient temperature (T _a) on ventilation and gas exchange in chukar partridges (Alectoris chukar) were determined after acclimation to low and high altitute (LA and HA; 340 and 3,800 m, respectively).At both LA and HA, oxygen consumption ( ) increased with decreasingT _a atT _a from 20 to –20°C. AtT _a of 35 to 40°C, increased above thermoneutral values at HA but remained constant and minimal at LA. Water loss rates increased rapidly atT _a>30°C at both altitudes as birds began to pant. Ventilation rates (f) during panting were 5-to 23-fold greater than the minimalf at thermoneutralT _a.Increased atT _a below thermoneutrality was supported by increased minute volume (V i) at both altitudes. The change inV i was primarily a function of changing tidal volume (V t), althoughf increased slightly asT _a declined. Oxygen extraction ( ) remained fairly constant atT _a below 20°C at both altitudes. BothV t and were considerably lower when birds were panting than at lowerT _a.Chukars showed few obvious ventilatory adaptations to HA. The 35% change in between 340 and 3,800 m was accommodated by a corresponding change inV i (btps), most of which was accomplished by increasedf at HA, along with a slight increase in .Abbreviations and symbols HA high altitude - LA low altitude - rate of evaporative water loss - oxygen extraction efficiency - f respiratory frequency - V t tidal volume - V i minute volume - BMR basal metabolic rate - MHP metabolic heat production     

Respiration of the emersed shore crab at variable ambient oxygenation

Summary In the intertidal shore crab,Carcinus maenas, pH and values of the prebranchial venous hemolymph, and , the PO₂ values of the arterialized cardiac hemolymph, , were measured, and the ventilatory activity was assessed by measuring the hydrostatic pressures at the exits of the epibranchial cavities, under four environmental conditions: normoxic water, normoxic air, hypoxic gas, hyperoxic gas.In the crab breathing normoxic air, was lower and higher than in animals breathing normoxic water. With the switch to hypoxic gas, ventilation increased and and decreased. With the switch to hyperoxic gas, ventilation decreased, and increased and decreased.Thus these crabs breathing air were not as well oxygenated as when breathing air-equilibrated water, and because of their low , they were relatively hypervetilating and hypocapnic compared to air-breathing vertebrates. Hyperoxic breathing, increasing and reducing ventilatory drive, led to increased . Conversely, was reduced by hypoxic breathing. These observations suggest that the gas exchanger of intertidal crabs is not as successfully designed for air breathing as that of land-colonizing insects and air-breathing vertebrates.