The length of discontinuous gas exchange cycles in lepidopteran pupae may serve as a mechanism for natural selection

Gas exchange is studied in diapausing pupae of Mamestra brassicae L., whose larvae are reared under identical conditions. The release of CO₂ gas is recorded with infrared gaseous analyzers. Oxygen convective uptake into the tracheae and oxygen consumption rates are recorded by means of a constant‐volume coulometric respirometer. Outputs from both of these respirometry systems are combined with infrared actographs. All 3‐month‐old pupae of M. brassicae display a pattern of discontinuous gas exchange (DGE) cycles of CO₂ gas release by bursts, although the lengths of these cycles varies between individuals. Some pupae exhibit long DGE cycles of at least 20 h in duration, with negligible CO₂ gas release during interburst periods, and there is presumed to be a convective gas exchange at this time. As a result of a partial vacuum inside the tracheae, a large oxygen convective uptake always occurs at the start of the spiracular opening phase. Other pupae have short DGE cycles of less than 3 h in duration, with elevated CO₂ gas release during the interburst period, when gas exchange is predominantly diffusive. The spiracular open phase in these pupae consists of frequent separate convective bursts of CO₂ gas release, with the opening–closing rhythms of the spiracles, which are considered as O phase fluttering. The pupae with long DGE cycles exhibit extremely low metabolic rates and very low total water loss rates, whereas those with short DGE cycles have higher metabolic and total water loss rates. The pupae with long DGE cycles live approximately twice as long as those with short cycles; thus, the present study demonstrates that long DGE cycles confer a fitness benefit on pupae as a result of a lower metabolic rate associated with water economy, conferring on them a longer life.     

Discontinuous gas exchange patterns of beet armyworm pupae, Spodoptera exigua (Lepidoptera: Noctuidae): effects of Bacillus thuringiensis Cry1C toxin, pupal age and temperature

Abstract. Changes in the discontinuous gas exchange cycle of pupal beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), exposed or not to Cry1C Bacillus thuringiensis toxin, are examined against developmental age (1–7 days) and at different temperatures (10–25 °C) using flow through respirometry. Both exposed and nonexposed pupae exhibit discontinuous gas exchange, but only at 10 °C; the frequency of cyclic release of CO₂ increases with increasing temperatures. The three phases of the discontinuous gas exchange cycle are distinct for both treatment groups. However, the duration of each phase is significantly greater for pupae exposed previously to toxin. The closed phase is 40 ± 14% longer, the flutter phase 23 ± 19% longer, and the open phase is 28 ± 12% longer when pupae were exposed to toxin. Respiratory water loss is 4.5 ± 1.3% for toxin exposed pupae and 2.1 ± 2.4% for unexposed pupae. Furthermore, the exposed pupae have significantly greater cuticular permeability (26.01 ± 1.9 µg cm⁻² h⁻¹ mmHg⁻¹) than the nonexposed pupae (9.64 ± 0.9 µg cm⁻² h⁻¹ mmHg⁻¹). However, in both strains, cuticular transpiration (>93%) far exceeds respiratory transpiration. Overall, total water loss is significantly greater in pupae whose larvae are exposed to toxin compared with pupae from nontreated larvae. Toxin exposed pupae have a mean cycle duration of 60 ± 2.5 min whereas that of nonexposed pupae is 42 ± 1.8 min.(ml g⁻¹ h⁻¹) of the open phase is greater earlier in pupal life followed by a minimum at mid-pupal stage and an increase at late-pupal development in both treatment groups. Combining all 7 days, closed, flutter and open phase (ml g⁻¹ h⁻¹), pupae exposed to toxin produce significantly more CO₂ during each phase. On average, toxin exposed pupae produce 52 ± 12, 43 ± 10 and 15 ± 37% more CO₂ than the untreated pupae during the closed, flutter and open phases, respectively. Therefore, the present study reinforces the need to use insects of similar developmental age in studies of insect respiration patterns and energy metabolism.     

The role of the spiracles in gas exchange during development of Samia cynthia (Lepidoptera, Saturniidae).

Spiracles and the tracheal system of insects allow effective delivery of respiratory gases. During development, holometabolous insects encounter large changes in the functional morphology of gas exchange structures. To investigate changes in respiratory patterns during development, CO2-release was measured in larvae, pre-pupae and pupae of Samia cynthia (Lepidoptera, Saturniidae). Gas exchange patterns showed great variability. Caterpillars had high metabolic rates and released carbon dioxide continuously. Pre-pupae and pupae showed typical discontinuous gas exchange cycles (DGC) at reduced metabolic rates. Changes in gas exchange patterns can partly be explained with low metabolic rates during pupation. Sequential blocking of spiracles in pre-pupae and pupae reduced spiracle conductance with tracheal conductance remaining unaffected. Analysis of gas exchange patterns indicates that caterpillars and pre-pupae use more than 14 spiracles simultaneously while pupae only use 8 to 10 spiracles. Total conductance is not a simple multiple of single spiracles, but may be gradually adaptable to gas exchange demands. Surprisingly, moth pupae showed a DGC if all except one spiracle were blocked. The huge conductance of single spiracles is discussed as a pre-adaptation to high metabolic demands at the beginning and the end of the pupal as well as in the adult stage.     

The role of the spiracles in gas exchange during development of Samia cynthia (Lepidoptera, Saturniidae)

Spiracles and the tracheal system of insects allow effective delivery of respiratory gases. During development, holometabolous insects encounter large changes in the functional morphology of gas exchange structures. To investigate changes in respiratory patterns during development, CO₂-release was measured in larvae, pre-pupae and pupae of Samia cynthia (Lepidoptera, Saturniidae). Gas exchange patterns showed great variability. Caterpillars had high metabolic rates and released carbon dioxide continuously. Pre-pupae and pupae showed typical discontinuous gas exchange cycles (DGC) at reduced metabolic rates. Changes in gas exchange patterns can partly be explained with low metabolic rates during pupation. Sequential blocking of spiracles in pre-pupae and pupae reduced spiracle conductance with tracheal conductance remaining unaffected. Analysis of gas exchange patterns indicates that caterpillars and pre-pupae use more than 14 spiracles simultaneously while pupae only use 8 to 10 spiracles. Total conductance is not a simple multiple of single spiracles, but may be gradually adaptable to gas exchange demands. Surprisingly, moth pupae showed a DGC if all except one spiracle were blocked. The huge conductance of single spiracles is discussed as a pre-adaptation to high metabolic demands at the beginning and the end of the pupal as well as in the adult stage.     

Discontinuous gas exchange, water loss, and metabolism in Protaetia cretica (Cetoniinae, Scarabaeidae)

Insects are at high risk of desiccation because of their small size, high surface-area-to-volume ratio, and air-filled tracheal system that ramifies throughout their bodies to transport O(2) and CO(2) to and from respiring cells. Although the tracheal system offers a high-conductance pathway for the movement of respiratory gases, it has the unintended consequence of allowing respiratory transpiration to the atmosphere. When resting, many species exchange respiratory gases discontinuously, and an early hypothesis for the origin of these discontinuous gas exchange cycles (DGCs) is that they serve to reduce respiratory water loss. In this study, we test this "hygric" hypothesis by comparing rates of CO(2) exchange and water loss among flower beetles Protaetia cretica (Cetoniinae, Scarabaeidae) breathing either continuously or discontinuously. We show that, consistent with the expectations of the hygric hypothesis, rates of total water loss are higher during continuous gas exchange than during discontinuous gas exchange and that the ratio of respiratory water loss to CO(2) exchange is lower during discontinuous gas exchange. This conclusion is in agreement with other studies of beetles and cockroaches that also support the hygric hypothesis. However, this result does not exclude other adaptive hypotheses supported by work on ants and moth pupae. This ambiguity may arise because there are multiple independent evolutionary origins of DGCs and no single adaptive function underlying their genesis. Alternatively, the observed reduction in water loss during DGCs may be a side effect of a nonadaptive gas exchange pattern that is elicited during periods of inactivity.     

Cyclic CO(2) emissions during the high temperature pulse of fluctuating thermal regime in eye-pigmented pupae of Megachile rotundata

Megachile rotundata (Hymenoptera: Megachilidae), the primary pollinator used in alfalfa seed production, may need to be exposed to low-temperature storage to slow the insects' development to better match spring emergence with the alfalfa bloom. It has been demonstrated that using a fluctuating thermal regime (FTR) improves the tolerance of pupae to low temperatures. Carbon dioxide emission rates were compared between four different FTRs, all with a base temperature of 6 °C and a daily high-temperature pulse. Four different high-temperature pulses were examined, 15 or 25 °C for 2 h and 20 °C for 1 or 2 h. A subset of pupae at the FTR base temperature of 6 °C exhibited continuous gas exchange and, once ramped to 20 or 25 °C, shifted to cyclic gas exchange. As temperatures were ramped down from the high-temperature pulse to 6 °C, the pupae reverted to continuous gas exchange. The following conclusions about the effect of FTR on the CO₂ emissions of M. rotundata pupae exposed to low-temperature storage during the spring incubation were reached: 1) the high temperature component of the FTR was the best predictor of respiratory pattern; 2) neither pupal body mass nor days in FTR significantly affected which respiratory pattern was expressed during FTRs; 3) cyclic gas exchange was induced only in pupae exposed to temperatures greater than 15 °C during the FTR high temperature pulse; and 4) a two hour pulse at 25 °C doubled the number of CO₂ peaks observed during the FTR pulse as compared to a two hour pulse at 20 °C.     

Effect of Cyclic Variations in Gas Exchange under Constant Environmental Conditions on the Ratio of Transpiration to Net Photosynthesis

Simultaneous cyclic variation in rates of both net photosynthesis and transpiration were induced in attached leaves of cotton and pepper plants under constant environmental conditions. The cyclic variations in photosynthesis and transpiration were found to be in phase, and the ratio net photosynthetic rate/transpiration rate remained constant over a wide range of gas exchange rates. A similar constancy of this ratio was also found as gas exchange rates declined following excision of a sunflower leaf, which was not initially cycling, in air. These results suggested that change in stomatal aperture was the only controlling factor involved and that it was affecting both processes proportionately. Visible loss of leaf turgur and measurable water stress developed in both pepper and cotton at peak exchange rates, but the gas exchange ratio remained constant. The failure of water stress and increased stomatal aperture to lower the gas exchange ratio suggested an absence of any significant leaf mesophyll resistance (r′_m) to inward diffusion of CO₂. The possibility that r′_m was low is discussed generally, and in relation to the use of chemical antitranspirants to raise the gas exchange ratio. Within the limits of the experiments, water stress apparently had no direct adverse effect on rates of net photosynthesis. The gas exchange ratio did not rise as exchange rates declined. Ultimately, at very low exchange rates, the ratio fell, declining to zero in cotton, but not in pepper. This decline was attributed to the onset of significant gas exchange through the cuticle, which was apparently less permeable to CO₂ than to water vapour. Positive net cuticular photosynthesis therefore probably does not occur in cotton. Except at very low exchange rates, the gas exchange ratio was higher in cotton than in pepper; it was similar in sunflower and cotton.     

昆虫不连续气体交换

许多昆虫呼吸时气体交换是不连续的循环式进行的。根据气门开闭,一个典型的不连续气体交换循环（discontinuous gas exchangecycle, DGC）可以明显分为3个阶段: 关闭阶段,极少或没有气体交换;颤动阶段,气门迅速微开和关闭,O₂进入气管,少量CO₂释放;最后是开放阶段,大量的CO₂释放。该文综述了DGC特征及昆虫活动、温度、体重对DGC的影响,并讨论了DGC与呼吸失水、缺氧或高CO₂浓度环境有关的进化适应意义。     

Effects of Neem EC on gas exchange, tracheal ventilation, and water loss in diapausing pupae of Pieris brassicae

Effects of Neem EC (The Indian Neem Tree Company^TM, 1% azadirachtin) on gas exchange cycles, tracheal ventilation, and water loss in diapausing pupae of the large white butterfly, Pieris brassicae L. (Lepidoptera: Pieridae), were studied using a constant volume respirometer combined with an infrared probe actograph. The non‐treated pupae displayed discontinuous gas exchange cycles (DGC) with a trend coinciding with the bursts of carbon dioxide (CO₂) release, active tracheal ventilation, and the heartbeat periods. Two independent forms of tracheal ventilation were observed, relatively vigorous abdominal shaking movements and weak abdominal pulsations. The ability to respond to mechanical excitation with abdominal movements was entirely lost on the 2nd day after treatments with Neem EC, and also a reduced tendency to use a DGC was observed. During 2–3 days after treatments, the DGCs and gas exchange microcycles were entirely lost, as was active ventilation. Before treatments, body mass loss, that is, water loss, was 0.6–0.9 mg g^?1 day^?1. After the treatments, water loss increased to 3–5 mg g^?1 day^?1. The pupae remained alive for 10–15 days after the treatments and died after having lost about 50% of their initial body mass. The absence of heartbeats measured during at least 4–5 h was the main criterion for ascertaining death of pupae. The results suggested that respiratory failures, that is, the loss of cyclic gas exchange, evoked by Neem EC were the primary cause of lethal desiccation. Thus, the hypothesis that the cyclic gas exchange is an adaptation for restricting water losses in insects was supported.     

Ozone concentration in leaf intercellular air spaces is close to zero

Transpiration and ozone uptake rates were measured simultaneously in sunflower leaves at different stomatal openings and various ozone concentrations. Ozone uptake rates were proportional to the ozone concentration up to 1500 nanoliters per liter. The leaf gas phase diffusion resistance (stomatal plus boundary layer) to water vapor was calculated and converted to the resistance to ozone multiplying it by the theoretical ratio of diffusion coefficients for water vapor and ozone in air (1.67). The ozone concentration in intercellular air spaces calculated from the ozone uptake rate and diffusion resistance to ozone scattered around zero. The ozone concentration in intercellular air spaces was measured directly by supplying ozone to the leaf from one side and measuring the equilibrium concentration above the other side, and it was found to be zero. The total leaf resistance to ozone was proportional to the gas phase resistance to water vapor with a coefficient of 1.68. It is concluded that ozone enters the leaf by diffusion through the stomata, and is rapidly decomposed in cell walls and plasmalemma.     

Calculation of CO2 gas phase diffusion in leaves and its relation to stomatal resistance

A new theory and experimental method was developed to measure the diffusion resistance to CO₂ in the gas phase of mesophyll leaf tissue. Excised leaves were placed in a chamber and their net evaporation and CO₂ assimilation rates measured at two different ambient pressures. These data were used to calculate CO₂ gas phase diffusion resistances. A variety of field grown leaves were tested and the effects of various experimental errors considered. Increasing the gas phase diffusion resistance decreased transpiration more than it decreased CO₂ assimilation. It was concluded that gas phase diffusion resistance associated with CO₂ assimilation may sometimes be 100 or 200 s·m^-1 greater than the resistance implied by transpiration rates. This may be due to longer path lengths for the CO₂ diffusion, constricted in places by the shape and arrangement of mesophyll cells.     

Role of nitrogen in transmucosal gas exchange rate in the rat middle ear.

This study investigates the role of nitrogen (N2) in transmucosal gas exchange of the middle ear (ME). We used an experimental rat model to measure gas volume variations in the ME cavity at constant pressure. We disturbed the steady-state gas composition with either air or N2 to measure resulting changes in volume at ambient pressure. Changes in gas volume over time could be characterized by three phases: a primary transient increase with time (phase I), followed by a linear decrease (phase II), and then a gradual decrease (phase III). The mean slope of phase II was -0.128 microl/min (SD 0.023) in the air group (n = 10) and -0.105 microl/min (SD 0.032) in the N2 group (n = 10), but the difference was not significant (P = 0.13), which suggests that the rate of gas loss can be attributed mainly to the same steady-state partial pressure gradient of N2 reached in this phase. Furthermore, a mathematical model was developed analyzing the transmucosal N2 exchange in phase II. The model takes gas diffusion into account, predicting that, in the absence of change in mucosal blood flow rate, gas volume in the ME should show a linear decrease with time after steady-state conditions and gas composition are established. In accordance with the experimental results, the mathematical model also suggested that transmucosal gas absorption of the rat ME during steady-state conditions is governed mainly by diffusive N2 exchange between the ME gas and its mucosal blood circulation.     

Spiracle activity in moth pupae—The role of oxygen and carbon dioxide revisited

After decades of intensive research, the actual mechanism behind discontinuous gas exchange in insects has not been fully understood. One open question concerns the actual way (closed, flutter, and open) of how spiracles respond to tracheal gas concentrations. As the results of a classic paper [Burkett, B.N., Schneiderman, H.A., 1974. Roles of oxygen and carbon dioxide in the control of spiracular function in cecropia pupae. Biological Bulletin 147, 274-293] allow ambiguous interpretation, we thus reexamined the behavior of the spiracles in response to fixed, controlled endotracheal gas concentrations.The tracheal system of diapausing pupae of Attacus atlas (Saturniidae, Lepidoptera) was flushed with gas mixtures varying in PO₂ and PCO₂ while the behavior of the spiracles was monitored using changes in the pressure signal. This novel pressure based technique proved to be superior to classic visual observation of single spiracles. A two-dimensional map of the spiracle behavior in response to endotracheal PO₂ and PCO₂ was established. Typically, it contained two distinct regions only, corresponding to “closed” and “open” spiracles. A separate “flutter” region was missing. Because fluttering is commonly observed in moth pupae, we suggest that the intermittent spiracle opening during a flutter phase is an effect of non-steady-state conditions within the tracheal system. For low PCO₂ the minimum PO₂ resulting in open spiracles was linearly dependent upon PCO₂. Above a threshold of 1-1.5 kPa CO₂ the spiracles were open irrespective of PO₂. We propose a hypothetical spiracular control model, which is simple and explains the time course of endotracheal partial pressures during all phases of discontinuous gas exchange.     

Stomata of the CAM plant Tillandsia recurvata respond directly to humidity

Summary Under controlled conditions, CO₂ exchange of Tillandsia recurvata showed all characteristics of CAM. During the phase of nocturnal CO₂ fixation stomata of the plant responded sensitively to changes in ambient air humidity. Dry air resulted in an increase, moist air in a decrease of diffusion resistance. The evaporative demand of the air affected the level of stomatal resistance during the entire night period. Due to stomatal closure, the total nocturnal water loss of T. recurvata was less at low than at high humidity. It is concluded that stomata respond directly to humidity and not via bulk tissue water conditions of the leaves. Such control of transpiration may optimize water use efficiency for this almost rootless, extreme epiphyte.     

Modulation of cyclic CO2 release in response to endogenous changes of metabolism during pupal development of Zophobas rugipes (Coleoptera: Tenebrionidae)

Understanding the mechanisms of gas exchange regulation in insects currently is a hot topic of insect physiology. Endogenous variation of metabolism during pupal development offers a great opportunity to study the regulation of respiratory patterns in insects. Here we show that metabolic rates during pupal development of the tenebrionid beetle Zophobas rugipes reveal a typical U-shaped curve and that, with the exception of 9-day-old pupae, the time between two bursts of CO₂ (interburst phase) was the only parameter of cyclic CO₂ gas exchange patterns that was adjusted to changing metabolic rates. The volume of CO₂ released in a burst was kept constant, suggesting a regulation for accumulation and release of a fixed amount of CO₂ throughout pupal development. We detected a variety of discontinuous and cyclic gas exchange patterns, which were not correlated with any periods of pupal development, suggesting a high among individual variability. An occasional occurrence of continuous CO₂ release patterns at low metabolic rates was very likely caused by single defective non-occluding spiracles.     

The use of the anaesthetic, enflurane, for determination of metabolic rates and respiratory parameters in insects, using the ant, Camponotus maculatus (Fabricius) as the model

This study investigated the effects of the anaesthetic, enflurane, on metabolic rates and ventilation patterns in the spotted sugar ant, Camponotus maculatus, using flow-through respirometry. The standard metabolic rate was not affected by the anaesthetic. While the ants were anaesthetised they exhibited a similar discontinuous gas exchange cycle to that observed when they were voluntarily motionless, but their spiracles remained open for a longer time during the open or burst phase even though the amount of CO(2) emitted during this phase remained constant. We discuss this finding in the context of the central nervous system control of the spiracle muscle. For both the determination of standard metabolic rate and ventilation patterns the individual ant has to be motionless. From this study we recommend the use of enflurane to ensure immobility in ants, and other small active insects, during the determination of standard metabolic rates, but the anaesthetic cannot be used to quantify the respiration pattern.     

Effect of a 5-HT1A receptor agonist (8-OH-DPAT) on external urethral sphincter activity in a rat model of pudendal nerve injury

Although serotonergic agents have been used to treat patients with stress urinary incontinence, the characteristics of the external urethral sphincter (EUS) activity activated by 5-HT receptors have not been extensively studied. This study examined the effects of the 5-HT(1A) receptor agonist, 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), on the EUS-electromyography and resistance of the urethra in a rat model with bilateral pudendal nerve injury (BPNI). Two measurements were utilized to assess the effects of the drug on bladder and urethral functions: the simultaneous recordings of transvesical pressure under isovolumetric conditions [isovolumetric intravesical pressure (IVP)] and urethral perfusion pressure, and the simultaneous recordings of IVP during continuously isotonic transvesical infusion with an open urethra (isotonic IVP) and EUS-electromyography. This study also evaluated the urethral continence using leak point pressure testing. The urethral perfusion pressure and leak point pressure measurements of BPNI rats reveal that 8-OH-DPAT significantly increased urethral resistance during the bladder storage phase, yet decreased resistance during the voiding phase. The entire EUS burst period was significantly prolonged, within which the average silent period increased and the frequency of burst discharges decreased. 8-OH-DPAT also improved the voiding efficiency, as evidenced by the detection of decreases in the contraction amplitude and residual volume, with increases in contraction duration and voided volume. These findings suggest that 8-OH-DPAT not only improved continence function, but also elevated the voiding function in a BPNI rat model.     

Analysis of acetylene reduction rates of soybean nodules at low acetylene concentrations

It has been previously proposed that acetylene reduction data at subsaturating acetylene concentrations could be interpreted by use of the Michaelis-Menten equation, based on the acetylene concentration external to the nodules. One difficulty of this view is that the assumption that the system is not diffusion limited is violated when studying intact nodules. The presence of a gas diffusion barrier in the nodule cortex leads to an alternate expression for the gas exchange rates at subsaturating gas concentrations. A theoretical comparison of the `apparent' Michaelis-Menten model and diffusion model illustrated the difficulties observed in the former model of overestimating the Michaelis-Menten coefficient and yielding a correlation between the Michaelis-Menten coefficient and the maximum rate. On the other hand, use of a diffusion model resulted in (a) estimates of the Michaelis-Menten coefficient consistent with enzyme studies, (b) stability of the estimates of the Michaelis-Menten coefficient independent of treatment, and (c) a sensitivity of the diffusion barrier conductance to plant drought stress. It was concluded that all studies of nodule gas exchange need to consider possible effects caused by the presence of a diffusion barrier.     

The pH mediated effects of initial glucose concentration on the transitory occurrence of extracellular metabolites, gas exchange and growth yields of aerobic batch cultures of Klebsiella pneumoniae

The changes in growth kinetics in aerobic batch cultures of Klebsiella pneumoniae were followed by measurements of extracellular metabolites, rates of gas exchange, dissolved oxygen tension, pH, and carbon balance at all stages of growth. When the initial growth-limiting glucose concentration in media without pH control was increased from 1.0 g carbon L(-1) to 2.2 g carbon L(-1), the number of different, mainly acidic, extracellular metabolites of glucose at the end of exponential growth increased, while the proportion of acetate decreased. During the postexponential growth phase, the extracellular metabolites were oxidized, resulting in an increasing complexity of changes in pH, gas exchange, and dissolved oxygen tension with increasing initial substrate concentration. All these parameters showed concomitant stepwise changes. This pattern was independent of the dissolved oxygen tension in the range 30-200 muM. When pH was kept constant, the number, slope, and relative magnitude of the steps in gas exchange and dissolved oxygen tension were pH-dependent, being most complex at low pH. Detailed carbon balances showed that 20% of the initial glucose was converted into extracellular metabolites at the end of exponential growth at neutral pH. In the postexponential phase, pyruvate (2%) was reoxidized first followed by acetate (13%). The observed molar growth yield coefficient (Y(ATP)) was 8.4 if the transitory occurrence of pyruvate and acetate was accounted for, and 6.4 if it was neglected. The corrected observed molar growth yield coefficient (Y'(ATP)) was 9.4 and compared well with the true molar growth yield coefficient (Y(Max) (ATP)), which was found to be 11.0. Specific in situ respiration rates of the exponential growth phase of cultures grown at different controlled pH values compared well with in situ values for energy-limited chemostat grown cells at the same growth rates, suggesting that growth in the batch culture was energy-limited throughout the exponential growth phase. This view was supported by low levels of intracellular glycogen and exopolysaccharides of all cultures, by the value of Y'(ATP) of 9.4, and by a constant specific production rate of the extracellular metabolites throughout exponential growth. It was concluded that even under strictly aerobic conditions, control of pH is as important as control of dissolved oxygen tension during growth of enterobacteriaceae in batch cultures.     

Environmental influences on the development of the cardiac system in fish and amphibians

In poikilothermic animals body temperature varies with environmental temperature, and this results in a change in metabolic activity (Q10 of enzymatic reactions typically is around 2-3). Temperature changes also modify gas transport in body fluids. While the diffusion coefficient increases with increasing temperatures, physical solubility and also hemoglobin oxygen affinity decrease. Therefore, an increase in temperature typically requires adjustments in cardiac activity because ventilatory and convectional transport of respiratory gases usually are tightly coupled in adults in order to meet the oxygen demand of body tissues. Hypoxic conditions also provoke adaptations in the central circulatory system, like the hypoxic bradycardia, which has been described for many adult lower vertebrates, combined with an increase in stroke volume and peripheral resistance. In embryos and larvae the situation is much more complicated, because nervous control of the heart is established only late during development, and because the site of gas exchange changes from mainly cutaneous gas exchange during early development to mainly pulmonary or branchial gas exchange in late stages. In addition, recent studies in amphibian and fish embryos and larvae reveal, that at least in very early stages convectional gas transport of the hemoglobin is not essential, which means that in these early stages ventilatory and convectional gas transport are not yet coupled. Accordingly, in early stages of fish and amphibians the central cardiac system often does not respond to hypoxia, although in some species behavioral adaptations indicate that oxygen sensors are functional. If a depression of cardiac activity is observed, it most likely is a direct effect of oxygen deficiency on the cardiac myocytes. Regulated cardiovascular responses to hypoxia appear only in late stages and are similar to those found in adult species.