Temperature effects on oxygen consumption and breathing pattern in juvenile and adult Chelonoidis carbonarius (Spix, 1824).

The effects of temperature on breathing pattern and oxygen consumption are being investigated in juvenile tortoises and compared to adults, in order to understand physiological adjustments of the respiratory system as related to body size, especially regarding the energetic expenditure associated with growth. We analyzed the breathing pattern and oxygen consumption of juvenile and adult red-footed tortoises (Chelonoidis carbonarius, Cryptodira: Testudinidae). The animals (N = 9; body mass ranging from 0.03 Kg to 2.5 Kg) were exposed to normoxic-normocarbic conditions using open respirometry in order to determine the breathing pattern and oxygen consumption in three different temperatures (15, 25, 35 °C). The obtained results showed intermittent breathing pattern in all tested temperatures in juveniles and adults. Tidal volume was not affected by changes in temperature, while breathing frequency increased significantly with increasing temperature, leading to a significant increment in minute ventilation between 15 and 35 °C. Mass specific oxygen consumption increased significantly with temperature and juveniles showed greater values when compared to adults. The alterations in the ventilatory response to temperature changes occurred in order to maintain the oxygen supply with increased metabolic activity. The differences between juveniles and adults in breathing frequency lead to juveniles needing a lower ventilation rate to perform gas exchange while extracting more oxygen. While these differences might be attributed to a greater metabolic expenditure during development, scaling effects on respiratory variables might be the main contributors to the found differences.

Baroreflex responses to activity at different temperatures in the South American rattlesnake, Crotalus durissus.

In humans, physical exercise imposes narrower limits for the heart rate (fH) response of the baroreflex, and vascular modulation becomes largely responsible for arterial pressure regulation. In undisturbed reptiles, the baroreflex-related fH alterations at the operating point (Gop) decreases at elevated body temperatures (Tb) and the vascular regulation changes accordingly. We investigated how the baroreflex of rattlesnakes, Crotalus durissus, is regulated during an activity at different Tb, expecting that activity would reduce the capacity of the cardiac baroreflex neural pathway to buffer arterial pressure fluctuations while being compensated by the vascular neural pathway regulation. Snakes were catheterized for blood pressure assessment at three different Tb: 15, 20 and 30 °C. Data were collected before and after activity at each Tb. Baroreflex gain (Gop) was assessed with the sequence method; the vascular limb, with the time constant of pressure decay (τ), using the two-element Windkessel equation. Both Gop and τ reduced when Tb increased. Activity also reduced Gop and τ in all Tb. The relationship between τ and pulse interval (τ/PI) was unaffected by the temperature at resting snakes, albeit it reduced after activity at 20 °C and 30 °C. The unchanged τ/PI and normalized Gop at different Tb indicated those variables are actively adjusted to work at different fH and pressure conditions at rest. Our data suggest that during activity, the baroreflex-related fH response is attenuated and hypertension is buffered by a disproportional increase in the rate which pressure decays during diastole. This compensation seems especially important at higher Tb where Gop is already low.

Effects of different levels of hypoxia and hypercarbia on ventilation and gas exchange in Boa constrictor amaralis and Crotalus durissus (Squamata: Serpentes).

Ventilation and gas exchange have been studied in relatively few species of snakes, especially regarding their response to environmental hypoxia or hypercarbia. We exposed Crotalus durissus (N = 6) and Boa constrictor (N = 6) to decreasing levels of oxygen (12, 9, 6, 3 % O2) and increasing levels of carbon dioxide (1.5, 3.0, 4.5, 6.0 % CO2) and analyzed the effect of the different gas mixtures on ventilation and gas exchange using open-flow respirometry. Neither hypoxia nor hypercarbia significantly altered the duration of expiration or inspiration, nor their proportions. Both hypoxia and hypercarbia increased minute ventilation, but the decrease in oxygen had a less pronounced effect on ventilation. Gas exchange under normoxic conditions was low and was not significantly affected by hypoxia, but hypercarbia decreased gas exchange significantly in both species. While B. constrictor maintained its respiratory exchange ratio (RER) under hypercarbia between 0.5 and 1.0, C. durissus showed a RER above 1.0 during hypercarbia, due to a significantly greater CO2 excretion. The overall responses of both species to hypercarbia and especially to hypoxia were very similar, which could be associated to similar lifestyles as ambush hunting sit-and-wait predators that are able to ingest large prey items. The observed differences in gas exchange could be related to respiratory systems with macroscopically different structures, possessing only a tracheal lung in C. durissus, but two functional lungs in B. constrictor.

Heart rate variability in the tegu lizard, Salvator merianae, its neuroanatomical basis and role in the assessment of recovery from experimental manipulation.

Using long-term, remote recordings of heart rate (fH) on fully recovered, undisturbed lizards, we identified several components of heart rate variability (HRV) associated with respiratory sinus arrhythmia (RSA): 1.) A peak in the spectral representation of HRV at the frequency range of ventilation. 2.) These cardiorespiratory interactions were shown to be dependent on the parasympathetic arm of the autonomic nervous system. 3.) Vagal preganglionic neurons are located in discrete groups located in the dorsal motor nucleus of the vagus and also, in a ventro-lateral group, homologous to the nucleus ambiguus of mammals. 4.) Myelinated nerve fibers in the cardiac vagus enabling rapid communication between the central nervous system and the heart. Furthermore, the study of the progressive recovery of fH in tegu following anesthesia and instrumentation revealed that 'resting' levels of mean fH and reestablishment of HRV occurred over different time courses. Accordingly, we suggest that, when an experiment is designed to study a physiological variable reliant on autonomic modulation at its normal, resting level, then postsurgical reestablishment of HRV should be considered as the index of full recovery, rather than mean fH.

Evaluation of the sequence method as a tool to assess spontaneous baroreflex in reptiles.

The sequence method is an alternative to the traditional pharmacological approach (i.e., the Oxford technique) used to calculate baroreflex gain (G) in mammals. Although the sequence method assesses baroreflex by measuring spontaneous events of blood pressure regulation, the pharmacological method relies on the injection of vasoactive drugs that impact the baroreflex mechanism itself. The sequence method might be relevant for dynamic measurement of baroreflex modulation but it was never validated for vertebrates with low heart rate. Hence, we tested the sequence method in three species of reptiles and compared the results with those provided by the classic pharmacological method. G was similar between both methods and values correlated when parameters for the sequence method were set at delay 0 or 1 (i.e., the baroreflex system responds immediately to blood pressure changes or after 1 heartbeat). Calculation of the baroreflex effectiveness index was adequate at a minimum of 300 cycles and a delay of 1 for the three species. Therefore, the sequence method has been validated to investigate baroreflex regulation in reptiles, enabling studies during dynamic alterations in homeostasis.

Convective oxygen transport during development in embryos of the snapping turtle Chelydra serpentina.

This study investigated the maturation of convective oxygen transport in embryos of the snapping turtle (Chelydra serpentina). Measurements included: mass, oxygen consumption (V̇O2 ), heart rate, blood oxygen content and affinity and blood flow distribution at 50%, 70% and 90% of the incubation period. Body mass increased exponentially, paralleled by increased cardiac mass and metabolic rate. Heart rate was constant from 50% to 70% incubation but was significantly reduced at 90% incubation. Hematocrit and hemoglobin concentration were constant at the three points of development studied but arteriovenous difference doubled from 50% to 90% incubation. Oxygen affinity was lower for the early 50% incubation group (stage 19) compared with all other age groups. Blood flow was directed predominantly to the embryo but was highest to the chorioallantoic membrane (CAM) at 70% incubation and was directed away from the yolk as it was depleted at 90% incubation. The findings indicate that the plateau or reduction in egg V̇O2  characteristic of the late incubation period of turtle embryos may be related to an overall reduction in mass-specific V̇O2  that is correlated with decreasing relative heart mass and plateaued CAM blood flow. Importantly, if the blood properties remain unchanged prior to hatching, as they did during the incubation period studied in the current investigation, this could account for the pattern of V̇O2 previously reported for embryonic snapping turtles prior to hatching.

Effects of environmental hypoxia and hypercarbia on ventilation and gas exchange in Testudines.

BACKGROUND: Ventilatory parameters have been investigated in several species of Testudines, but few species have had their ventilatory pattern fully characterized by presenting all variables necessary to understand changes in breathing pattern seen under varying environmental conditions. METHODS: We measured ventilation and gas exchange at 25 °C in the semi-aquatic turtle Trachemys scripta and the terrestrial tortoise Chelonoidis carbonarius under normoxia, hypoxia, and hypercarbia and furthermore compiled respiratory data of testudine species from the literature to analyze the relative changes in each variable. RESULTS: During normoxia both species studied showed an episodic breathing pattern with two to three breaths per episode, but the non-ventilatory periods (TNVP) were three to four times longer in T. scripta than in C. carbonarius. Hypoxia and hypercarbia significantly increased ventilation in both species and decreased TNVP and oxygen consumption in T. scripta but not in C. carbonarius. DISCUSSION: Contrary to expectations, the breathing pattern in C. carbonarius did show considerable non-ventilatory periods with more than one breath per breathing episode, and the breathing pattern in T. scripta was found to diverge significantly from predictions based on mechanical analyses of the respiratory system. A quantitative analysis of the literature showed that relative changes in the ventilatory patterns of chelonians in response to hypoxia and hyperbarbia were qualitatively similar among species, although there were variations in the magnitude of change.

The autonomic control of upright posture tachycardia in the arboreal lizard Iguana iguana.

In terrestrial environments, upright spatial orientation can dramatically influence animals' hemodynamics. Generally, large and elongated species are particularly sensitive to such influence due to the greater extent of their vascular beds being verticalized, favoring the establishment of blood columns in their bodies along with caudal blood pooling, and thus jeopardizing blood circulation through a cascade effect of reductions in venous return, cardiac filling, stroke volume, cardiac output, and arterial blood pressure. This hypotension triggers an orthostatic-(baroreflex)-tachycardia to normalize arterial pressure, and despite the extensive observation of this heart rate (fH ) adjustment in experiments on orthostasis, little is known about its mediation and importance in ectothermic vertebrates. In addition, most of the knowledge on this subject comes from studies on snakes. Thus, our objective was to expand the knowledge on this issue by investigating it in an arboreal lizard (Iguana iguana). To do so, we analyzed fH , cardiac autonomic tones, and fH variability in horizontalized and tilted iguanas (0°, 30°. and 60°) before and after muscarinic blockade with atropine and double autonomic blockade with atropine and propranolol. The results revealed that I. Iguana exhibits significant orthostatic-tachycardia only at 60o inclinations-a condition that is primarily elicited by a withdrawal of vagal drive. Also, as in humans, increases in low-frequency fH oscillations and decreases in high-frequency fH oscillations were observed along with orthostatic-tachycardia, suggesting that the mediation of this fH adjustment may be evolutionarily conserved in vertebrates.

The consequences of seasonal fasting during the dormancy of tegu lizards (Salvator merianae) on their postprandial metabolic response.

Tegu lizards (Salvator merianae) aestivate for up to 5 months during Brazil's winter, when they retreat to burrows and halt most activities. Dormant tegus reduce their gastrointestinal (GI) mass, which allows a substantial energy economy. This strategy, however, implies that the first post-dormancy digestion would be more costly than subsequent feeding episodes as a result of GI atrophy. To address this, we determined the postprandial metabolic response (SDA) of the first (M1), second (M2) and several (RM) feeding episodes after tegus' dormancy. Another group of tegus (PF) was subjected to an extra 50 day fasting period after arousal. Glucose, triglycerides and uric acid levels were checked before and after feeding. M1 digestion lasted twice as long and cost twofold more when compared with M2 or RM, in agreement with the idea that GI atrophy inflates digestion cost at the first post-dormancy meal. The SDA response was similar in M2 and RM, suggesting that the GI tract was fully reorganized after the first feeding. The SDA cost was equal in PF and RM, implying that the change in state per se (dormant to arousal) triggers the regrowth of GI, independently of feeding. Fasting tegus at M1 presented higher triglyceride and lower uric acid levels than fed tegus, indicating that fasting is mainly sustained by fat storage. Our results show that seasonal fasting imposes an extra digestion cost to tegus following their next feeding, which is fully paid during their first digestion. This surplus cost, however, is negligible compared with the overall energetic savings from GI tract atrophy during the dormancy period.

The Effects of Feeding on Cardiac Control of the Broad-Nosed Caiman (Caiman latirostris): The Role of the Autonomic Nervous System and NANC Factors.

The metabolic increment that occurs after feeding demands cardiovascular adjustments to be maintained, as increased heart rate (fH ) and cardiac output. In mammals, postprandial tachycardia seems to be triggered by an increase in adrenergic activity and by nonadrenergic noncholinergic (NANC) factors, while in ectothermic vertebrates, this adjustment seems to be linked to a withdrawal of vagal drive as well as to NANC factors. Because the factors behind postprandial tachycardia have not yet been investigated in crocodilians, the present study sought to evaluate the postprandial tachycardia mediators in the broad-nosed caiman. To this end, fasting and digesting animals were instrumented with intraperitoneal cannula and subcutaneous electrocardiogram electrodes (for the measurement of fH , cardiac autonomic tones, and total fH variability, as well as for a power spectral analysis of fH ). Data were then collected with the animals in an untreated state, as well as after muscarinic cholinergic blockade with atropine (2.5 mg kg-1 ) and after double autonomic blockade with atropine and propranolol (5.0 mg kg-1 ). Fasting animals' fH was ∼18 bpm, a value which increased to ∼30 bpm during digestion. After the double autonomic blockade, fasting animals exhibited an fH of ∼15 bpm, while digesting animals' fH was ∼23 bpm. This result is evidence of the presence of NANC factors with positive chronotropic effects acting during digestion. The calculated autonomic tones showed that, after feeding, the adrenergic tone increased while the cholinergic tone remained unchanged. Finally, fH variability analyses revealed that this adrenergic increase is primarily derived from circulating catecholamines.

Vagal tone regulates cardiac shunts during activity and at low temperatures in the South American rattlesnake, Crotalus durissus.

The undivided ventricle of non-crocodilian reptiles allows for intracardiac admixture of oxygen-poor and oxygen-rich blood returning via the atria from the systemic circuit and the lungs. The distribution of blood flow between the systemic and pulmonary circuits may vary, based on differences between systemic and pulmonary vascular conductances. The South American rattlesnake, Crotalus durissus, has a single pulmonary artery, innervated by the left vagus. Activity in this nerve controls pulmonary conductance so that left vagotomy abolishes this control. Experimental left vagotomy to abolish cardiac shunting had no effect on long-term survival and failed to identify a functional role in determining metabolic rate, growth or resistance to food deprivation. Accordingly, the present investigation sought to evaluate the extent to which cardiac shunt patterns are actively controlled during changes in body temperature and activity levels. We compared hemodynamic parameters between intact and left-vagotomized rattlesnakes held at different temperatures and subjected to enforced physical activity. Increased temperature and enforced activity raised heart rate, cardiac output, pulmonary and systemic blood flow in both groups, but net cardiac shunt was reversed in the vagotomized group at lower temperatures. We conclude that vagal control of pulmonary conductance is an active mechanism regulating cardiac shunts in C. durissus.

Beyond body size: muscle biochemistry and body shape explain ontogenetic variation of anti-predatory behaviour in the lizard Salvator merianae.

Anti-predatory behaviour evolves under the strong action of natural selection because the success of individuals avoiding predation essentially defines their fitness. Choice of anti-predatory strategies is defined by prey characteristics as well as environmental temperature. An additional dimension often relegated in this multilevel equation is the ontogenetic component. In the tegu Salvator merianae, adults run away from predators at high temperatures but prefer fighting when it is cold, whereas juveniles exhibit the same flight strategy within a wide thermal range. Here, we integrate physiology and morphology to understand ontogenetic variation in the temperature-dependent shift of anti-predatory behaviour in these lizards. We compiled data for body shape and size, and quantified enzyme activity in hindlimb and head muscles, testing the hypothesis that morphophysiological models explain ontogenetic variation in behavioural associations. Our prediction is that juveniles exhibit body shape and muscle biochemistry that enhance flight strategies. We identified biochemical differences between muscles mainly in the LDH:CS ratio, whereby hindlimb muscles were more glycolytic than the jaw musculature. Juveniles, which often use evasive strategies to avoid predation, have more glycolytic hindlimb muscles and are much smaller when compared with adults 1-2 years old. Ontogenetic differences in body shape were identified but marginally contributed to behavioural variation between juvenile and adult tegus, and variation in anti-predatory behaviour in these lizards resides mainly in associations between body size and muscle biochemistry. Our results are discussed in the ecological context of predator avoidance by individuals differing in body size living at temperature-variable environments, where restrictions imposed by the cold could be compensated by specific phenotypes.

Seasonal reproductive endothermy in tegu lizards.

With some notable exceptions, small ectothermic vertebrates are incapable of endogenously sustaining a body temperature substantially above ambient temperature. This view was challenged by our observations of nighttime body temperatures sustained well above ambient (up to 10°C) during the reproductive season in tegu lizards (~2 kg). This led us to hypothesize that tegus have an enhanced capacity to augment heat production and heat conservation. Increased metabolic rates and decreased thermal conductance are the same mechanisms involved in body temperature regulation in those vertebrates traditionally acknowledged as "true endotherms": the birds and mammals. The appreciation that a modern ectotherm the size of the earliest mammals can sustain an elevated body temperature through metabolic rates approaching that of endotherms enlightens the debate over endothermy origins, providing support for the parental care model of endothermy, but not for the assimilation capacity model of endothermy. It also indicates that, contrary to prevailing notions, ectotherms can engage in facultative endothermy, providing a physiological analog in the evolutionary transition to true endothermy.

Ventilation and gas exchange in two turtles: Podocnemis unifilis and Phrynops geoffroanus (Testudines: Pleurodira).

Turtles (Testudines) have two major taxa, Pleurodira and Cryptodira. To date, only limited data are available regarding the respiratory physiology of pleurodirans. To begin to address this, we studied ventilation and gas exchange in Podocnemis unifilis and Phrynops geoffroanus. Breathing pattern in both species could be described as episodic with breathing episodes separated by large non-ventilatory periods. We measured duration of inspiration and expiration, breathing frequency, duration of the non-ventilatory period (time between episodes), tidal volume, and oxygen consumption when breathing normoxia, hypoxia and hypercarbia at 25°C. In both species hypercarbia caused a greater increase in ventilation compared to hypoxia, increasing both breathing frequency and tidal volume. Minute ventilation and oxygen consumption in P. geoffroanus were the lowest described so far in testudines, indicating either extra-pulmonary gas exchange or a significantly lower metabolism. Oxidative costs of breathing, estimated using the regression method, was the highest described so far for any reptile. Further studies are necessary to better understand respiratory physiology in Phrynops and Podocnemis species.

Autonomic regulation of heart rate during specific dynamic action associated with digestion in the bullfrog Lithobates catesbeianus

Feeding causes a substantial rise in the aerobic metabolism of all vertebrates, which is met by a rise in heart rate to secure adequate oxygen delivery, as well as intestinal nutrient absorption and transport between organs. To study the autonomic regulation of this postprandial tachycardia in an amphibian, we placed arterial catheters in individuals of the bullfrog, Lithobates catesbeianus (Shaw, 1802), to measure their blood pressure and heart rate. Heart rate in experimental subjects increased from a fasting value of 42.5 ± 2.5 to 52.5 ± 1.6 and 57.5 ± 2.1 min-1 at 24 and 33 hours, respectively, after meals that corresponded to 5% of their body weight. Using an infusion of atropine and propranolol, we demonstrated that part of the postprandial tachycardia can be ascribed to a progressive decrease in parasympathetic tone (26 ± 5% in fasting frogs to 6 ± 5% by 33 hours into digestion), whereas the sympathetic tone remained unaltered at approximately 20%. In addition to the withdrawal of vagal tone, digestion was also associated with an increase in the heart rate upon double-blockade (i.e., the combination of atropine and propranolol) from a fasting value of 44.5 ± 0.9 to 50.8 ± 1.9 min-1 at 33 hours. This indicates that the postprandial tachycardia in frogs, as previously demonstrated for snakes, is partially governed by excitation of non adrenergic, non cholinergic (NANC) factors.

Autonomic regulation of heart rate during specific dynamic action associated with digestion in the bullfrog Lithobates catesbeianus

Feeding causes a substantial rise in the aerobic metabolism of all vertebrates, which is met by a rise in heart rate to secure adequate oxygen delivery, as well as intestinal nutrient absorption and transport between organs. To study the autonomic regulation of this postprandial tachycardia in an amphibian, we placed arterial catheters in individuals of the bullfrog, Lithobates catesbeianus (Shaw, 1802), to measure their blood pressure and heart rate. Heart rate in experimental subjects increased from a fasting value of 42.5 ± 2.5 to 52.5 ± 1.6 and 57.5 ± 2.1 min-1 at 24 and 33 hours, respectively, after meals that corresponded to 5% of their body weight. Using an infusion of atropine and propranolol, we demonstrated that part of the postprandial tachycardia can be ascribed to a progressive decrease in parasympathetic tone (26 ± 5% in fasting frogs to 6 ± 5% by 33 hours into digestion), whereas the sympathetic tone remained unaltered at approximately 20%. In addition to the withdrawal of vagal tone, digestion was also associated with an increase in the heart rate upon double-blockade (i.e., the combination of atropine and propranolol) from a fasting value of 44.5 ± 0.9 to 50.8 ± 1.9 min-1 at 33 hours. This indicates that the postprandial tachycardia in frogs, as previously demonstrated for snakes, is partially governed by excitation of non adrenergic, non cholinergic (NANC) factors.(AU)

Viabilidade econômica da criação de matrizes e reprodutores de Caiman latirostris no estado de São Paulo / Economic viability of Caiman latirostris in the state of São Paulo (Brazil): sale of matrices and reproducers

The breeding of crocodilians has great economic potential. The present study investigated the economic feasibility of crocodilians in the state of São Paulo in different scenarios (sale 40-100 animals per year), with the goal of producing matrices and reproducers for renewal or formation of breeding in other creations, through a sensitivity and stochastic analysis (Monte Carlo simulation). The results of the sensitivity analysis show favorability of this activity in a 15-year horizon, with a Net Present Value (NPV) of R$ -202,804.18. However, the projection of variables for a risk analysis (stochastic) enabled the survival of the crocodilian farm without major changes and lower costs for farmers, revealing a low-risk activity, indicating a high probability of economic return.(AU)

A criação de crocodilianos possui grande potencial econômico. O presente estudo investigou a viabilidade econômica da criação de crocodilianos no estado de São Paulo em diferentes cenários (venda de 40 a 100 animais ao ano), com o objetivo de produzir matrizes e reprodutores para a renovação/formação de plantel de outras criações, através de análise de sensibilidade e também estocástica (simulação de Monte Carlo). Os resultados da análise de sensibilidade não mostraram favorabilidade desta atividade num horizonte de 15 anos, apresentando Valor Presente Líquido (VPL) de R$ -202.804,18. Contudo, a projeção de variáveis para uma análise de riscos (estocástica) possibilitou a sobrevivência da criação de crocodilianos sem grandes mudanças e baixos custos para o produtor rural, revelando uma atividade de baixo risco e apontando a probabilidade elevada de retorno econômico.(AU)

Viabilidade econômica da criação de matrizes e reprodutores de Caiman latirostris no estado de São Paulo / Economic viability of Caiman latirostris in the state of São Paulo (Brazil): sale of matrices and reproducers

The breeding of crocodilians has great economic potential. The present study investigated the economic feasibility of crocodilians in the state of São Paulo in different scenarios (sale 40-100 animals per year), with the goal of producing matrices and reproducers for renewal or formation of breeding in other creations, through a sensitivity and stochastic analysis (Monte Carlo simulation). The results of the sensitivity analysis show favorability of this activity in a 15-year horizon, with a Net Present Value (NPV) of R$ -202,804.18. However, the projection of variables for a risk analysis (stochastic) enabled the survival of the crocodilian farm without major changes and lower costs for farmers, revealing a low-risk activity, indicating a high probability of economic return.

A criação de crocodilianos possui grande potencial econômico. O presente estudo investigou a viabilidade econômica da criação de crocodilianos no estado de São Paulo em diferentes cenários (venda de 40 a 100 animais ao ano), com o objetivo de produzir matrizes e reprodutores para a renovação/formação de plantel de outras criações, através de análise de sensibilidade e também estocástica (simulação de Monte Carlo). Os resultados da análise de sensibilidade não mostraram favorabilidade desta atividade num horizonte de 15 anos, apresentando Valor Presente Líquido (VPL) de R$ -202.804,18. Contudo, a projeção de variáveis para uma análise de riscos (estocástica) possibilitou a sobrevivência da criação de crocodilianos sem grandes mudanças e baixos custos para o produtor rural, revelando uma atividade de baixo risco e apontando a probabilidade elevada de retorno econômico.

An appraisal of the use of an infrared digital monitoring system for long-term measurement of heart rate in reptilian embryos.

Measurement of heart rate (fH) in embryonic reptiles has previously imposed some degree of invasive treatment on the developing embryo. Recently a non-invasive technique of fH detection from intact eggs was developed for commercial avian breeders and has since been used in biological research. This device uses infrared light, enabling it to detect heartbeats in very early embryos. However, infrared light is a source of heat and extended enclosure of an egg in the device is likely to affect temperature with consequent effects on physiological processes, including fH. We studied the effect of use of the monitor on the temperature of eggs and on fH in two species of reptiles, the snapping turtle (Chelydra serpentina) and the green iguana (Iguana iguana). Egg temperature increased from a room temperature of 27-28 °C, by 26% in turtles and 14% in iguanas over 1h of enclosure, resulting in an increase in fH of 76-81% in turtles and 35-50% iguanas. These effects on fH can either be avoided by brief enclosure of each egg in the monitor or measured and accounted for during the design of long-term experiments.

The progressive onset of cholinergic and adrenergic control of heart rate during development in the green iguana, Iguana iguana.

The autonomic control of heart rate was studied throughout development in embryos of the green iguana, Iguana iguana by applying receptor agonists and antagonists of the parasympathetic and sympathetic systems. Acetylcholine (Ach) slowed or stopped the heart and atropine antagonized the response to Ach indicating the presence of muscarinic cholinoceptors on the heart of early embryos. However, atropine injections had no impact on heart rate until immediately before hatching, when it increased heart rate by 15%. This cholinergic tonus increased to 34% in hatchlings and dropped to 24% in adult iguanas. Although epinephrine was without effect, injection of propranolol slowed the heart throughout development, indicating the presence of ß-adrenergic receptors on the heart of early embryos, possibly stimulated by high levels of circulating catecholamines. The calculated excitatory tonus varied between 33% and 68% until immediately before hatching when it fell to 25% and 29%, a level retained in hatchlings and adults. Hypoxia caused a bradycardia in early embryos that was unaffected by injection of atropine indicating that hypoxia has a direct effect upon the heart. In later embryos and hatchlings hypoxia caused a tachycardia that was unaffected by injection of atropine. Subsequent injection of propranolol reduced heart rate both uncovering a hypoxic bradycardia in late embryos and abolishing tachycardia in hatchlings. Hypercapnia was without effect on heart rate in late stage embryos and in hatchlings.