Effects of temperature and hypercapnia on ventilation and breathing pattern in the lizard Uromastyx aegyptius microlepis

In most reptiles, the ventilatory response to hypercapnia consists of large increases in tidal volume (V(T)), whereas the effects on breathing frequency (f(R)) are more variable. The increased V(T) seems to arise from direct inhibition of pulmonary stretch receptors. Most reptiles also exhibit a transitory increase in ventilation upon removal of CO(2) and this post-hypercapnic hyperpnea may consist of changes in both V(T) and f(R). While it is well established that increased body temperature augments the ventilatory response to hypercapnia, the effects of temperature on the post-hypercapnic hyperpnea is less described. In the present study, we characterise the ventilatory response of the agamid lizard Uromastyx aegyptius to hypercapnia and upon the return to air at 25 and 35 degrees C. At both temperatures, hypercapnia caused large increases in V(T) and small reductions in f(R), that were most pronounced at the higher temperature. The post-hypercapnic hyperpnea, which mainly consisted of increased f(R), was numerically larger at 35 compared to 25 degrees C. However, when expressed as a proportion of the levels of ventilation reached during steady-state hypercapnia, the post-hypercapnic hyperpnea was largest at 25 degrees C. Some individuals exhibited buccal pumping where each expiratory thoracic breath was followed by numerous small forced inhalations caused by contractions of the buccal cavity. This breathing pattern was most pronounced during severe hypercapnia and particularly evident during the post-hypercapnic hyperpnea.     

The ventilatory response to environmental hypercarbia in the South American rattlesnake,<Emphasis Type="Italic"> Crotalus durissus</Emphasis>

To study the effects of environmental hypercarbia on ventilation in snakes, particularly the anomalous hyperpnea that is seen when CO₂ is removed from inspired gas mixtures (post-hypercapnic hyperpnea), gas mixtures of varying concentrations of CO₂ were administered to South American rattlesnakes, Crotalus durissus, breathing through an intact respiratory system or via a tracheal cannula by-passing the upper airways. Exposure to environmental hypercarbia at increasing levels, up to 7% CO₂, produced a progressive decrease in breathing frequency and increase in tidal volume. The net result was that total ventilation increased modestly, up to 5% CO₂ and then declined slightly on 7% CO₂. On return to breathing air there was an immediate but transient increase in breathing frequency and a further increase in tidal volume that produced a marked overshoot in ventilation. The magnitude of this post-hypercapnic hyperpnea was proportional to the level of previously inspired CO₂. Administration of CO₂ to the lungs alone produced effects that were identical to administration to both lungs and upper airways and this effect was removed by vagotomy. Administration of CO₂ to the upper airways alone was without effect. Systemic injection of boluses of CO₂-rich blood produced an immediate increase in both breathing frequency and tidal volume. These data indicate that the post-hypercapnic hyperpnea resulted from the removal of inhibitory inputs from pulmonary receptors and suggest that while the ventilatory response to environmental hypercarbia in this species is a result of conflicting inputs from different receptor groups, this does not include input from upper airway receptors.Communicated by G. Heldmaier     

Patterns of intermittent ventilation and responses to perfusing gas mixtures in quiescent Blaberus craniifer

ABSTRACT. Patterns of intermittent ventilation were recorded by means of long electromyogram wires from quiescent Blaberus craniifer (Burmeister) buried in vermiculite. While buried, cockroaches were subjected to perfusion with various mixtures of CO₂ in air and of oxygen in nitrogen. Quiescent cockroaches in air ventilated for mean periods of 138 s in cycles of 720s duration, but much variability occurred within and between cockroaches. Mild hypercapnia or hypoxia shortened the overall cycle time while more severe treatment caused the cycle to be replaced by continual pumping. Intermittent ventilation persisted in decapitated insects but the threshold of the response to hypoxia or hypercapnia was elevated. Prevailing gas tensions normally determine the frequency and duration of each phase.     

Oxygen and acid-base status related drives to gill ventilation in carp

Gill ventilation in carp Cyprinus carpio increased 50% with a reduction of [O²]^a to 60% of the control value. Small decreases of inspired PO² also increased ventilation, which occurred without any significant changes of the blood gas picture. Ventilation increased with hypercapnia but only if [O²]^a became reduced by concomitant Bohr and Root shifts. Consistently, hypercapnia failed to increase ventilation when combined with hyperoxia. Therefore, it was not possible to ascribe any ventilatory response to specific CO²/pH receptors.     

Respiratory and acid-base disturbances in rainbow trout blood during exposure to chloramine-T under hypoxia and hyperoxia

Rainbow trout Oncorhynchus mykiss were exposed acutely to chloramine-T at a therapeutic concentration (9mg l⁻¹) under moderately hypoxic (water P o₂: l00 mmHg) or hyperoxic (water P o₂: 430mmHg) conditions and arterial blood gas tensions ( P ao₂ and P aco₂) and pH were monitored using an extracorporeal circulation. Hypoxia, alone, resulted in an increased ventilation frequency, a decrease in both arterial P co₂ and P o₂ and an increase in arterial pH. There was no effect of chloramine-T exposure on the measured variables as compared with pre-exposure (hypoxia baseline) values. Hyperoxia, alone, resulted in a decrease in ventilation frequency, an increase in arterial P co₂ and P o₂, and a decrease in arterial pH. Chloramine-T exposure under these conditions caused a significant increase in ventilation frequency, but no significant effect on arterial blood gases or pH as compared with the hyperoxia baseline values. Despite the increase in ventilation caused by chloramine-T during hyperoxia, there was no reduction in P co₂ or increase in P o₂. Although these results were of little pathological significance, this study suggests that chloramine-T, although stimulating ventilation, was impairing the diffusion of co₂ across the gill probably by the secretion of branchial mucus and enhancing the gill boundary layer.     

Measurements of photosynthesis and respiration in plants

Methods for measuring the rates of photosynthesis and respiration in plants are reviewed. Closed systems that involve manometric techniques, ¹⁴CO₂ fixation, O₂ electrodes and other methods for measuring dissolved and gas phase O₂ are described. These methods typically provide time-integrated rate measurements, and limitations to their use are discussed. Open gas exchange systems that use infra-red CO₂ gas analysers and differential O₂ analysers for measuring instantaneous rates of CO₂ and O₂ exchange are described. Important features of the analysers, design features of gas exchange systems, and sources of potential error are considered. The analysis of chlorophyll fluorescence parameters for estimating the quantum yield for O₂ evolution and CO₂ fixation is described in relation to new fluorescence imaging systems for large scale screening of photosynthetic phenotypes, and the microimaging of individual chloroplasts.     

Rhythms of passive and active ventilation, and circulation recorded in diapausing pupae of Mamestra brassicae using constant volume respirometry

Abstract.  The periodically occurring convective inflow of air into the tracheal system, or passive suction ventilation, together with the cyclic bursts of release of CO₂ and active ventilation, is recorded in diapausing pupae of Mamestra brassicae . A constant volume respirometer combined with an opto-cardiograph-actograph is used. In all pupae with a metabolic rate of 0.025–0.054 mL g⁻¹ h⁻¹, the bouts of almost imperceptible abdominal contractions are recorded during the bursts of carbon dioxide release and this mode of active ventilation is qualified as extracardiac haemocoelic pulsations. The pupae whose metabolic rate is 0.052–0.075 mL⁻¹ g⁻¹ h⁻¹ show more vigorous abdominal contractions. The results demonstrate that, in diapausing pupae, characterized with low metabolic rates, both passive suction ventilation, referred to also as passive suction inspiration, and active ventilation occurs. In approximately 50% of the pupae, each gas exchange microcycle during the interburst periods begins with a miniature PSI followed by a microburst of CO₂ release; in approximately 30% of the individuals, passive suction inspirations occur separately from CO₂ microbursts; in the remaining pupae, miniature ones without microbursts of CO₂ are recorded. A typical event is heartbeat reversion: in longer periods, the heart peristalses are directed forward (anterograde of heartbeat) and, in shorter periods, the heart peristalses are directed backward (retrograde of heartbeat). At 0 °C, the cyclic release of CO₂ and miniature passive suction inspirations during the interburst periods are preserved at lower frequencies but active ventilation is lost.     

Control of ventilatory movements in the aquatic insect Corydalus comutus: central effect of hypoxia

ABSTRACT. The influence of hypoxia and hypercapnia on the ventilatory rhythm of the hellgrammite Corydalus cornutus L. (Megaloptera) was studied. In intact animals the frequency of rhythmic retractions and protractions of abdominal gills is increased by hypoxia (10% O₂, 90% N₂) but no ventilatory response is elicited by hypercapnia (1–5% CO₂, 20% O₂, 75–79% N₂).
The ventilatory motor pattern was examined by recording extracellularly from the gill retractor muscle or its efferent nerve. In response to hypoxia (8% 0₂, 92% N₂), there are decreases in the cycle-time, the interspike interval, and the burst length of the gill retractor motorneuron. In addition, previously quiescent motorneurons associated with gill protraction are recruited.
Individual ganglia or small groups of abdominal ganglia can be isolated both from the central ganglionic chain and from the periphery by selective cutting of roots and connectives. When exposed to hypoxia, preparations that include the first abdominal ganglion show characteristic changes in the ventilatory motor pattern similar to those in intact animals. Thus sensitivity to oxygen appears to be located centrally and not peripherally. In small animals (head width < 7 mm), abdominal ganglia 2–3 and 2–7 respond characteristically to hypoxia, but in larger animals (head width > 9 mm), chains of ganglia lacking abdominal ganglion 1 fail to respond. In larger animals oxygen sensitivity may thus be concentrated in abdominal ganglion 1, whereas in smaller animals the ability to initiate a ventilatory response to hypoxia is distributed among the abdominal ganglia.     

Influence of freshwater sediment on the survival of Escherichia coli and Salmonella sp. as measured by three methods of enumeration

The thermophilic hydrogen-oxidizing eubacterium, Hydrogenobacter thermophilus was grown with a semi-continuous gas feed in a 150 1 fermentor, on different ratios of H₂, O₂ (air) and CO₂ and on media of variable mineral salts composition. The best gas ratio during the exponential phase was approximately 0.5:1.0:0.03 atm of H₂: air: CO₂. Cell densities were increased by increasing the concentration of ammonium and phosphate in the medium. The maximum cell density obtained under the most favourable conditions was 2.62 g wet wt 1^-1, corresponding to an absorption of 1.6 at 600 nm. The maximum specific growth rate ( u ) was 0.44 h^-1.     

A Chromatographic Method for Analysis of the Gaseous Phase in Shake Flask Cultures and its Application to the Study of Methane Utilization

A simple gas chromatograph with a katharometer detector is described to determine O₂, N₂, methane and CO₂ in gas samples of 0·01–2·0 ml. The apparatus is inexpensive, and can be modified to determine other gases. The sensitivity to oxygen is 3 × 10⁻⁶ g. The use of the instrument is illustrated by a study of the growth kinetics of Methylococcus capsulatus grown on methane in shake flask experiments. The ratio of O₂ uptake to methane uptake is much lower in the stationary phase than in the growth phase of the culture.     

Acid-base regulation in the South American lungfish Lepidosiren paradoxa: effects of prolonged hypercarbia on blood gases and pulmonary ventilation

The South American lungfish (Lepidosiren paradoxa) has well-developed lungs and highly reduced gills. To evaluate acid-base regulation, we applied hypercarbia while blood gases and pulmonary ventilation were measured for up to 48 h. Dorsal aortic blood was analyzed, and pulmonary ventilation was measured by pneumotachography. Two protocols were used: (1) normocarbia (control) followed by aquatic hypercarbia (7% CO2 approximately 49 mmHg), gas phase normocarbic; and (2) normocarbia (control) followed by combined aquatic/gas phase hypercarbia (7% CO2). Normocarbic values were pHa~7.5, Paco2 approximately 17 mmHg, and [HCO-3]pl approximately 22 mM. For protocol 1, the first hour of exposure increased Paco2 from 17.0 to 37.4 mmHg, and pHa fell to 7.21 and remained there for the rest of the experiment. At 3 h, pulmonary ventilation reached sixfold the normocarbic value but then decreased. For protocol 2, combined gas phase/water hypercarbia had a large effect on acid-base status. Thus, Paco2 increased gradually to 74 mmHg (pHa=7.15) at 48 h. At 3 h, ventilation reached a sixfold increase relative to normocarbic control but then rose further to a 60-fold peak at 6 h, followed by a gradual decline. As in some salamanders and air-breathing teleosts, there was no evidence of active extracellular modulation bicarbonate.     

Detection and identification of gibberellins in extracts of Begonia leaves by bioassay, radioimmunoassay and gas chromatography – mass spectrometry

Gibberellins A₁ (GA₁), GA₄, GA₉, GA₁₉, and GA₂₀ were identified in extracts of leaves of Begonia x cheimantha Everett cv. Nova (Christmas or Lorraine Begonia). GA-like substances were purified by reverse phase and normal phase high performance liquid chromatography (HPLC) and detected by Tan-ginbozu dwarf rice bioassay and binding to antibodies raised against GA₁, GA₄ and GA₉. The final identifications were made by gas chromatography—mass spectrometry (GC-MS).     

THE EFFECT OF MODERATE AND MARKED HYPERCAPNIA UPON THE ENERGY STATE AND UPON THE CYTOPLASMIC NADH/NAD+ RATIO OF THE RAT BRAIN

Abstract— The energy state of brain tissue was evaluated from the tissue concentrations of ATP, ADP and AMP and the cytoplasmic NADH/NAD⁺ ratio from the tissue, CSF and blood concentrations of lactate and pyruvate, and from the intracellular pH', in rats exposed to carbon dioxide concentrations of 640 per cent. The hypercapnia had no significant effect on the energy state of the tissue. Hypercapnia of increasing severity gave rise to a progressive decrease in the pyruvate concentration; the lactate concentration fell at low CO₂ concentrations, but no further decrease was observed at CO₂ concentrations greater than 20 per cent. There was a progressive rise in the intracellular lactate/pyruvate ratio at increasing CO₂ concentrations, corresponding to the fall in intracellular pH, i.e. the calculated NADH/NAD⁺ ratios remained normal. It is therefore concluded that hypercapnia does not affect the cytoplasmic redox state.     

Acetogenesis from H2 and CO2 by methane- and non-methane-producing human colonic bacterial communities

Abstract: The purpose of the study was to define the potential for reductive acetogenesis of colonic microflora from six non-methane- and four methane-excreting human subjects in relation to numbers of the different H₂-utilizing microorganisms. Faecal bacterial suspensions were incubated in the presence of NaH¹³CO₃ and under a gas phase composed of either 100% N₂ (control) or 80% H₂–20% N₂. The effects of a specific methanogenesis inhibitor or of sulfate supplementation were also determined. Quantitative nuclear magnetic resonance showed the presence of both single- and double-labelled acetate in all incubations under hydrogen. H₂/CO₂-acetogenesis appears to be a quantitatively important activity only in the presence of very low numbers of methanogens. Inhibition of methanogenesis induced a large increase in ¹³CO₂ incorporation into acetate in CH₄-producing samples. These results showed that methanogens can efficiently outcompete acetogens in human colonic contents. In contrast, no clear-cut competition for H₂ between acetogenesis and dissimilatory sulfate-reduction could be demonstrated. A slight reduction of the acetogenic activity was only observed at the highest sulfate addition (100 mM).     

The effects of acute hypoxia and hypercapnia on oxygen consumption of the freshwater European eel

When exposed to hypoxia, eels Anguilla anguilla were able to regulate and maintain Vo₂ down to a water oxygen tension ( Pwo₂ ) of about 25 mmHg, a value far below those reported in other studies. When exposed to hypercapnia, eels showed a depression in Vo₂ as water carbon dioxide tension ( Pwco₂ ) increased. Faced with combined hypoxia-hypercapnia, eels showed an increase in their sensitivity to hypoxia, and the critical oxygen tension increased to 40–45 mmHg. The possible mechanisms underlying these responses were discussed, and the implications of such findings for extensive culture of eels were highlighted.     

Effects of hypercapnia on the buffer capacity and haematological values in Salmo salar (L.)

The effects of high environmental carbon dioxide tension (PCO₂) on buffering status, pH, haematocrit, red blood cell (RBC) count, mean corpuscle volume (MCV), and chloride (Cl^–) distribution in the blood of young salmon were investigated after 30 min and 24 h. An increase in blood buffering capacity was observed after 24 h. As compared to the preacclimation situation, haematocrit, RBC, MCV and Cl^– content of the erythrocytes increased after 30 min but decreased if hypercapnia was sustained for 24 h. However the plasma bicarbonate (HCO₃) content increased over the whole experimental period. Plasma Cl^– content was decreased after 24 h, but the ratio of Cl^– between erythrocytes and plasma increased at the onset of hypercapnia, then remained unchanged during the rest of the investigation period.     

The Measurement of Spinal Cord Tissue pH Using a Diffusible, Lipid-Soluble, pH-Sensitive Fluorescent Indicator

Abstract: Spinal cord tissue pH was measured in cats at normocapnia, hypocapnia, hypercapnia and death from anoxia using a pH-sensitive fluorescent indicator (umbelliferone) with both molecular and ionic fluorophors. A ratio analysis of the indicator's calibrated 450 nm fluorescent tissue clearance curves from 340 and 370 nm excitation permitted direct in vivo tissue pH determinations. Fifteen animals were divided into three equal groups according to different arterial carbon dioxide tensions (P_a co₂):five animals at P_a co₂ 20, five animals P_a co₂ 40 and five animals P_a co₂ 60 torr. Spinal cord tissue pH varied linearly with arterial pH, but within narrower limits. These values (arterial versus cord pH) were: 7.46 versus 7.15; 7.21, 7.09; and 7.04, 7.00. At death from hypoxemia the arterial pH fell to 6.99 and spinal cord to 6.67. The clearance curves of umbelliferone in spinal cord varied according to P_a co₂ and appeared to reflect spinal cord blood flow.     

Measurement of denitrification in estuarine sediment using membrane inlet mass spectrometry

Abstract Denitrification was measured in intact sediment cores and in homogenised slurries using membrane inlet mass spectrometry. Dissolved concentrations of O₂, N₂, N₂O and CO₂ were simultaneously monitored. Using a 0.8 mm diameter needle probe, a comparison was made of the gas profiles of intact cores obtained under different conditions, i.e. with air or argon as the headspace gas and after the addition of nitrate and/or a carbon source to the sediment surface. O₂ was detectable to a depth of 1 cm under a headspace of air and the depth at which the maxima of denitrification products occurred was 1.5–2 cm. Denitrification products (N₂O, N₂) occurred in the surface layers where O₂ was above the minimum level of detectability (> 0.25 μM): diffusion of N₂ and N₂O upwards from the anoxic zone, local anaerobic microenvironments or aerobic denitrification are alternative explanations for this observation. The addition of nitrate and/or acetate increased the concentrations of N₂, N₂O and CO₂ in the sediment core. In sediment slurries, the pH, nitrate concentration, carbon source and the depth from which the sample was taken affected the rate of denitrification. Nitrogen was the sole detectable end product. Maximum denitrification occurred at pH 7.5 and at 20 mM nitrate. Denitrification was at a maximum in those slurries prepared from sections of core at 1–2 cm depth.     

Respiratory responses to hypoxia in relation to mode of life of two erythrinid species (Hoplias malabaricus and Hoplias lacerdae)

Predominantly, Hoplias malabaricus inhabits stagnant O₂ poor environments, whereas Hoplias lacerdae occurs in well-aerated streams. The present study evaluates the influence of mode of life on O₂ uptake and gill ventilation in equally-sized (300 g) specimens of this genus at 25° C. Comparing the species, H. lacerdae was characterized by the highest O₂ uptake and gill ventilation combined with a relatively higher cost of breathing and a lower O₂ extraction. Both species substantially increased ventilation in response to hypoxia with the difference that H. malabaricus exclusively augmented tidal volume, whereas H. lacerdae also increased breathing frequency.     

The influence of leaf thickness on the CO2 transfer conductance and leaf stable carbon isotope ratio for some evergreen tree species in Japanese warm-temperate forests

1. The influence of leaf thickness on internal conductance for CO₂ transfer from substomatal cavity to chloroplast stroma ( g _i) and carbon isotope ratio (δ¹³C) of leaf dry matter was investigated for some evergreen tree species from Japanese temperate forests. g _i was estimated based on the combined measurements of gas exchange and concurrent carbon isotope discrimination.
2. Leaves with thicker mesophyll tended to have larger leaf dry mass per area (LMA), larger surface area of mesophyll cells exposed to intercellular air spaces per unit leaf area ( S _mes) and smaller volume ratio of intercellular spaces to the whole mesophyll (mesophyll porosity).
3. g _i of these leaves was correlated positively to S _mes but negatively to mesophyll porosity. The variation in g _i among these species would be therefore primarily determined by variation of the conductance in liquid phase rather than that in gas phase.
4. δ¹³C was positively correlated to mesophyll thickness and leaf nitrogen content on an area basis. However, g _i values did not correlate to δ¹³C. These results suggest that difference in δ¹³C among the species was not caused by the variation in g _i, but mainly by the difference in long-term photosynthetic capacity.
5. Comparison of our results with those of previous studies showed that the correlation between leaf thickness and g _i differed depending on leaf functional types (evergreen, deciduous or annual). Differences in leaf properties among these functional types were discussed.