Obesity prolongs induction times in reptiles.

Obesity is common in captive reptiles, and reptiles are increasingly popular as companion animals and in physiological research. Obesity may present a challenge during surgical procedures using inhalation anaesthesia, as the long induction time due to the low reptilian metabolism may increase anaesthetic accumulation in the adipose tissues. This study investigated the impact of obesity on induction and recovery times from inhaled anaesthesia. The temporal change in the partial pressure of isoflurane in different tissues was predicted using a multi-compartment model. Furthermore, as right-to-left shunting can delay anaesthetic uptake and washout, we included an assessment of the combination of cardiac shunting and obesity. The model predictions indicate a clear increase in time to reach 90% equilibration of administered anaesthetic in the brain (T90) of obese non-shunting (lean 47 min, obese >100 min) and shunting (lean 81 min, obese >100 min) reptiles. The combination of obesity and shunting doubled the time to acquisition of mean anaesthetic concentration (a measure used to plan anaesthesia) from 8 min to 19 min. Adipose blood flow highly affected whether the body type had an impact on induction time, with low adipose blood flow abolishing the effect of body type. As T90 was never reached within 100 min with both the obese reptiles, it was not possible to conclude on the effect of obesity on recovery times within this study. Care should therefore be taken when anaesthetising obese reptiles for surgical purposes, to ensure adequate anaesthetic depth is attained, and recovery monitored closely.

Ectothermy and cardiac shunts profoundly slow the equilibration of inhaled anaesthetics in a multi-compartment model.

The use of inhalational anaesthesia is ubiquitous in terrestrial vertebrates. Given the dependence of these agents on delivery by the cardiorespiratory system, we developed a new computational model predicting equilibration of inhaled anaesthetics in mammalian and ectotherm conditions including the ability of reptiles to maintain vascular shunts. A multi-compartment model was constructed from simultaneously-solved equations, verified by comparison to the literature for endo and ectotherm physiology. The time to 90% equilibration of anaesthetic in arterial blood (t90) is predicted and used to compare anaesthetics and physiologies. The five to tenfold lower cardiac output and minute ventilation of ectothermic vertebrates is predicted to slow equilibration times by five to ten times leading to 90% equilibration in ectotherm arterial blood of over 200 min, compounded by reduction in body temperature, and the extent of right-to-left vascular shunts. The impact of these findings is also influenced by the solubility coefficient of the anaesthetic, such that at net right-to-left shunt fractions of over 0.8, sevoflurane loses the advantage of faster equilibration, in comparison with isoflurane. We explore clinical strategies to regulate anaesthetic uptake in ectotherms by managing convectional flow especially by supportive ventilation and reduction of the right-to-left shunt.

The effects of morphine on gas exchange, ventilation pattern and ventilatory responses to hypercapnia and hypoxia in dwarf caiman (Paleosuchus palpebrosus).

Morphine and other opioids cause respiratory depression in high doses and lower the ventilatory responses to hypoxia and hypercapnia in mammals. Recent studies indicate that turtles respond similarly, but although they are used routinely for post-surgical analgesia, little is known about the physiological effects of opioids in reptiles. We therefore investigated the effects of morphine (10 and 20 mg kg-1) on gas exchange and ventilation in six dwarf caiman (Paleosuchus palpebrosus) using pneumotachography in a crossover design. Intraperitoneal injections of morphine changed the ventilation pattern from a typical intermittent/periodic pattern with a few or several breaths in ventilatory bouts to single breaths and prolonged the apnoea, such that respiratory frequency was depressed, while tidal volume was elevated. Furthermore, the duration of inspiration and especially expiration was prolonged. The resulting decrease in minute ventilation was attended by a lowering of the respiratory exchange ratio (RER) (especially for 20 mg kg-1 dose) indicating CO2 retention with a long time constant for approaching the new steady state. The changes in ventilation pattern and gas exchange reached a new stable level approximately 3 h after the morphine injection and did not significantly affect steady state O2 uptake, i.e. O2 consumption. As expected, the ventilatory response to 5% O2 was lower in morphine-treated caimans, but minute ventilation upon exposure to 2% CO2 did not differ significantly different from control animals.

Comparing anesthesia with isoflurane and fentanyl/fluanisone/midazolam in a rat model of cardiac arrest.

Only one in ten patients survives cardiac arrest (CA), underscoring the need to improve CA management. Isoflurane has shown cardio- and neuroprotective effects in animal models of ischemia-reperfusion injury. Therefore, the beneficial effect of isoflurane should be tested in an experimental CA model. We hypothesize that isoflurane anesthesia improves short-term outcome following resuscitation from CA compared with a subcutaneous fentanyl/fluanisone/midazolam anesthesia. Male Sprague-Dawley rats were randomized to anesthesia with isoflurane (n = 11) or fentanyl/fluanisone/midazolam (n = 11). After 10 min of asphyxial CA, animals were resuscitated by mechanical chest compressions, ventilations, and epinephrine and observed for 30 min. Hemodynamics, including coronary perfusion pressure, systemic O2 consumption, and arterial blood gases, were recorded throughout the study. Plasma samples for endothelin-1 and cathecolamines were drawn before and after CA. Compared with fentanyl/fluanisone/midazolam anesthesia, isoflurane resulted in a shorter time to return of spontaneous circulation (ROSC), less use of epinephrine, increased coronary perfusion pressure during cardiopulmonary resusitation, higher mean arterial pressure post-ROSC, increased plasma levels of endothelin-1, and decreased levels of epinephrine. The choice of anesthesia did not affect ROSC rate or systemic O2 consumption. Isoflurane reduces time to ROSC, increases coronary perfusion pressure, and improves hemodynamic function, all of which are important parameters in CA models.NEW & NOTEWORTHY The preconditioning effect of volatile anesthetics in studies of ischemia-reperfusion injury has been demonstrated in several studies. This study shows the importance of anesthesia in experimental cardiac arrest studies as isoflurane raised coronary perfusion pressure during resuscitation, reduced time to return of spontaneous circulation, and increased arterial blood pressure in the post-cardiac arrest period. These effects on key outcome measures in cardiac arrest research are important in the interpretation of results from animal studies.

Right-to-left shunt has modest effects on CO2 delivery to the gut during digestion, but compromises oxygen delivery.

By virtue of their cardiovascular anatomy, reptiles and amphibians can shunt blood away from the pulmonary or systemic circuits, but the functional role of this characteristic trait remains unclear. It has been suggested that right-to-left (R-L) shunt (recirculation of systemic blood within the body) fuels the gastric mucosa with acidified and CO2-rich blood to facilitate gastric acid secretion during digestion. However, in addition to elevating PCO2 , R-L shunt also reduces arterial O2 levels and would compromise O2 delivery during the increased metabolic state of digestion. Conversely, arterial PCO2  can also be elevated by lowering ventilation relative to metabolism (i.e. reducing the air convection requirement, ACR). Based on a mathematical analysis of the relative roles of ACR and R-L shunt on O2 and CO2 levels, we predict that ventilatory modifications are much more effective for gastric CO2 supply with only modest effects on O2 delivery. Conversely, elevating CO2 levels by means of R-L shunt would come at a cost of significant reductions in O2 levels. The different effects of altering ACR and R-L shunt on O2 and CO2 levels are explained by the differences in the effective blood capacitance coefficients.

The long road to steady state in gas exchange: metabolic and ventilatory responses to hypercapnia and hypoxia in Cuvier's dwarf caiman.

Animals with intermittent lung ventilation and those exposed to hypoxia and hypercapnia will experience fluctuations in the bodily O2 and CO2 stores, but the magnitude and duration of these changes are not well understood amongst ectotherms. Using the changes in the respiratory exchange ratio (RER; CO2 excretion divided by O2 uptake) as a proxy for changes in bodily gas stores, we quantified time constants in response to hypoxia and hypercapnia in Cuvier's dwarf caiman. We confirm distinct and prolonged changes in RER during and after exposure to hypoxia or hypercapnia. Gas exchange transients were evaluated in reference to predictions from a two-compartment model of CO2 exchange to quantify the effects of the levels of hypoxia and hypercapnia, duration of hypercapnia (30-300 min) and body temperature (23 versus 33°C). For hypercapnia, the transients could be adequately fitted by two-phase exponential functions, and slow time constants (after 300 min hypercapnia) concurred reasonably well with modelling predictions. The slow time constants for the decays after hypercapnia were not affected by the level of hypercapnia, but they increased (especially at 23°C) with exposure time, possibly indicating a temporal and slow recruitment of tissues for CO2 storage. In contrast to modelling predictions, elevated body temperature did not reduce the time constants, probably reflecting similar ventilation rates in transients at 23 and 33°C. Our study reveals that attainment of steady state for gas exchange requires considerable time and this has important implications for designing experimental protocols when studying ventilatory control and conducting respirometry.

Periodic ventilation: Consequences for the bodily CO2 stores and gas exchange efficiency.

Using a mathematical model of CO2 transport, we investigated the underlying cause of why and to what extent periodic ventilation is less efficient for CO2 excretion/elimination compared to continuous/tidal ventilation leading to elevated CO2 stores unless mean alveolar minute ventilation () is elevated. The model predicts that the reduced efficiency of periodic ventilation is intrinsic to the sequential arrangement and differences in the relative storage capacities (product of size and CO2 capacitance coefficient) of the lungs, blood and tissues that leads to predominant blood and tissue storage during apnoeic periods. Consequently, overall CO2 transport becomes more prone to perfusion and diffusion limitation during periodic ventilation. At constant cardiac output (Q.) inefficiency will increase with the apnoeic duration (tap) concomitant with increasing blood and tissues CO2 storage and with the relative time spent apnoeic (tap/tcyc) due to increasing V.A/Q. mismatch. Conversely, temporal variation of Q. to better match V.A can reduce inefficiency radically. Thus such adjustment in blood flow is necessary for efficient CO2 elimination in periodic ventilation.

Low cost of gastric acid secretion during digestion in ball pythons.

Due to their large metabolic responses to digestion (specific dynamic action, SDA), snakes represent an interesting animal group to identify the underlying mechanisms for the postprandial rise in metabolism. The SDA response results from the energetic costs of many different processes ranging over prey handling, secretions by the digestive system, synthesis of enzymes, plasticity of most visceral organs, as well as protein synthesis and nitrogen excretion. The contribution of the individual mechanisms, however, remains elusive. Gastric acid secretion has been proposed to account for more than half of the SDA response, while other studies report much lower contributions of the gastric processes. To investigate the energetic cost of gastric acid secretion, ball pythons (Python regius) were fed meals with added amounts of bone meal (up to 25 g bone meal kg(-1) snake) to achieve a five-fold rise in the buffer capacity of the meals. Direct measurements within the stomach lumen showed similar reduction in gastric pH when buffer capacity was increased, but we found no effects on the rise in oxygen consumption over the first three days of digestion. There was, however, a slower return of oxygen consumption to resting baseline. We conclude that gastric acid secretion only contributes modestly to the SDA response and propose that post-absorptive processes, such as increased protein synthesis, are likely to underlie the SDA response.

Closed system respirometry may underestimate tissue gas exchange and bias the respiratory exchange ratio (RER).

Closed respirometry is a commonly used method to measure gas exchange in animals due to its apparent simplicity. Typically, the rates of O2 uptake and CO2 excretion (VO2 and VCO2, respectively) are assumed to be in steady state, such that the measured rates of gas exchange equal those at tissue level. In other words, the respiratory gas exchange ratio (RER) is assumed to equal the respiratory quotient (RQ). However, because the gas concentrations change progressively during closure, the animal inspires air with a progressively increasing CO2 concentration and decreasing O2 concentration. These changes will eventually affect gas exchange causing the O2 and CO2 stores within the animal to change. Because of the higher solubility/capacitance of CO2 in the tissues of the body, VCO2 will be more affected than VO2, and we hypothesize therefore that RER will become progressively underestimated as closure time is prolonged. This hypothesis was addressed by a combination of experimental studies involving closed respirometry on ball pythons (Python regius) as well as mathematical models of gas exchange. We show that increased closed duration of the respirometer reduces RER by up to 13%, and these findings may explain previous reports of RER values being below 0.7. Our model reveals that the maximally possible reduction in RER is determined by the storage capacity of the body for CO2 (product of size and specific capacitance) relative to the respirometer storage capacity. Furthermore, modeling also shows that pronounced ventilatory and circulatory response to hypercapnia can alleviate the reduction in RER.

A critical evaluation of automated blood gas measurements in comparative respiratory physiology.

Precise measurements of blood gases and pH are of pivotal importance to respiratory physiology. However, the traditional electrodes that could be calibrated and maintained at the same temperature as the experimental animal are increasingly being replaced by new automated blood gas analyzers. These are typically designed for clinical use and automatically heat the blood sample to 37°C for measurements. While most blood gas analyzers allow for temperature corrections of the measurements, the underlying algorithms are based on temperature-effects for human blood, and any discrepancies in the temperature dependency between the blood sample from a given species and human samples will bias measurements. In this study we review the effects of temperature on blood gases and pH and evaluate the performance of an automated blood gas analyzer (GEM Premier 3500). Whole blood obtained from pythons and freshwater turtles was equilibrated in rotating Eschweiler tonometers to a variety of known P(O2)'s and P(CO2)'s in gas mixtures prepared by Wösthoff gas mixing pumps and blood samples were measured immediately on the GEM Premier 3500. The pH measurements were compared to measurements using a Radiometer BMS glass capillary pH electrode kept and calibrated at the experimental temperature. We show that while the blood gas analyzer provides reliable temperature-corrections for P(CO2) and pH, P(O2) measurements were substantially biased. This was in agreement with the theoretical considerations and emphasizes the need for critical calibrations/corrections when using automated blood gas analyzers.

Episodic ventilation lowers the efficiency of pulmonary CO2 excretion.

The ventilation pattern of many ectothermic vertebrates, as well as hibernating and diving endotherms, is episodic where breaths are clustered in bouts interspersed among apneas of varying duration. Using mechanically ventilated, anesthetized freshwater turtles (Trachemys scripta), a species that normally exhibits this episodic ventilation pattern, we investigated whether episodic ventilation affects pulmonary gas exchange compared with evenly spaced breaths. In two separate series of experiments (a noninvasive and an invasive), ventilation pattern was switched from a steady state, with evenly spaced breaths, to episodic ventilation while maintaining overall minute ventilation (30 ml·min(-1)·kg(-1)). On switching to an episodic ventilation pattern of 10 clustered breaths, mean CO2 excretion rate was reduced by 6 ± 5% (noninvasive protocol) or 20 ± 8% (invasive protocol) in the first ventilation pattern cycle, along with a reduction in the respiratory exchange ratio. O2 uptake was either not affected or increased in the first ventilation pattern cycle, while neither heart rate nor overall pulmonary blood flow was significantly affected by the ventilation patterns. The results confirm that, for a given minute ventilation, episodic ventilation is intrinsically less efficient for CO2 excretion, thereby indicating an increase in the total bodily CO2 store in the protocol. Despite the apparent CO2 retention, mean arterial Pco2 only increased 1 Torr during the episodic ventilation pattern, which was concomitant with a possible reduction of respiratory quotient. This would indicate a shift in metabolism such that less CO2 is produced when the efficiency of excretion is reduced.