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Development of the respiratory system can be affected by the use of drugs during pregnancy, as the prenatal phase is highly sensitive to pharmacological interventions, resulting in long-term consequences. The deleterious effects of external cannabinoids during gestation may be related to negative interference in central nervous system formation, cardiorespiratory system function, and behavioral disorders. Nevertheless, the impact of external cannabinoids on cardiorespiratory network development, chemosensitivity, and its future consequences in adulthood is still unclear. We evaluated the effects of prenatal exposure to a synthetic cannabinoid (WIN 55,212-2, 0.5 mg·kg-1·day-1) on the cardiorespiratory control and panic-like behavior of male and female rats in adulthood. Exogenous cannabinoid exposure during pregnancy resulted in a sex-dependent difference in breathing control. Specifically, males showed increased chemosensitivity to CO2 and O2, whereas females exhibited decreased sensitivity. Altered cardiovascular control was evident, with prenatally treated males and females being more susceptible to hypertension and tachycardia under adverse environmental conditions. Moreover, WIN-treated males exhibited higher fragmentation of sleep episodes, whereas females displayed anxiolytic and panicolytic behavioral responses to CO2. However, no changes were observed in the mechanical component of the respiratory system, and there were no neuroanatomical alterations, such as changes in the expression of CB1 receptors in the brainstem or in the quantification of catecholaminergic and serotonergic neurons. These findings highlight that external interference in cannabinoid signaling during fetal development causes sex-specific, long-lasting effects for the cardiorespiratory system and behavioral responses in adulthood.NEW & NOTEWORTHY The surge in recreational cannabis use and cannabinoid-based medication prescription among pregnant women has been notable in recent years, fueled by the misconception that natural products are inherently safe. Significant gaps persist regarding the potential risks of maternal consumption of cannabinoids and the long-term effects on the cardiorespiratory system of their offspring, which may be determined by sex. Accordingly, this research aims to diminish this lack of information and raise a note of caution.

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Teiid lizards possess an incomplete post-hepatic septum (PHS) separating the lungs and liver from the remaining viscera, and within this group, Salvator merianae has the most complete PHS. In this study, we explored the combined effects of the presence of the PHS and alterations in abdominal volume on the mechanics of the respiratory system. The PHS is believed to act as a mechanical barrier, mitigating the impact of the viscera on the lungs. Using established protocols, we determined static (Cstat) and dynamic (Cdyn) compliance, lung volume and work of breathing for the respiratory system in tegu lizards with intact (PHS+) or removed (PHS-) PHS, combined with (balloon+) or without (balloon-) increased abdominal volume. The removal of the PHS significantly reduced resting lung volume and Cdyn, as well as significantly increasing the work of breathing. An increase in abdominal volume significantly reduced Cstat, Cdyn, and resting and maximum lung volume. However, the work of breathing increased less in the PHS+/balloon+ treatment than in the PHS- treatments. These results highlight the barrier function of the PHS within the tegu lizard's body cavity. The septum effectively reduces the impact of the viscera on the respiratory system, enabling the lungs to be ventilated at a low work level, even when abdominal volume is increased. The presence of the PHS in teiid lizards underscores how extrapulmonary structures, such as septal divisions of the body cavity, can profoundly affect pulmonary breathing mechanics.

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Water is of fundamental importance to living organisms, not only as a universal solvent to maintain metabolic activity but also due to the effects the physical properties of water have on different organismal structures. In this review, we explore some examples of how living organisms deal with surfaces covered with or in contact with water. While we do not intend to describe all possible forms of interactions in every minute detail, we would like to draw attention to this intriguing interdisciplinary subject and discuss the positive and negative effects of the interaction forces between water molecules and organisms. Topics explored include locomotion on water, wettability of surfaces, benefits of retaining a film of air while submerged (Salvinia effect), surface tension of water inhibiting air-breathing, accumulation of water in small tubes, surface tension in non-mammalian and mammalian respiratory systems. In each topic, we address the importance of interactions with water and the adaptations seen in an organism to solve the surface-related challenges, trying to explore the different selective pressures acting onto different organisms allowing exploring or compensating these surface-related interactions.

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BACKGROUND AND PURPOSE: Cannabis legalization has risen in many countries, and its use during pregnancy has increased. The endocannabinoid system is present in the CNS at early stages of embryonic development, and regulates functional brain maturation including areas responsible for respiratory control, data on the influence of external cannabinoids on the development of the respiratory system and possible consequences during postnatal life are limited. EXPERIMENTAL APPROACH: We evaluated the effects of prenatal exposure to synthetic cannabinoid (WIN 55,212-2 [WIN], 0.5 mg·kg-1 ·day-1 ) on the respiratory control system in neonatal (P0, P6-7 and P12-13) and juvenile (P27-28) male and female rats. KEY RESULTS: WIN administration to pregnant rats interfered sex-specifically with breathing regulation of offspring, promoting a greater sensitivity to CO2 at all ages in males (except P6-7) and in juvenile females. An altered hypoxic chemoreflex was observed in P0 (hyperventilation) and P6-7 (hypoventilation) males, which was absent in females. Along with breathing alterations, brainstem analysis showed an increase in the number of catecholaminergic neurons and cannabinoid receptor type 1 (CB1 ) and changes in tissue respiration in the early males. A reduction in pulmonary compliance was observed in juvenile male rats. Preexposure to WIN enhanced spontaneous apnoea and reduced the number of serotoninergic (5-HT) neurons in the raphe magnus nucleus of P0 females. CONCLUSIONS AND IMPLICATIONS: These data demonstrate that excess stimulation of the endocannabinoid system during gestation has prolonged and sex-specific consequences for the respiratory control system.

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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.

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ABSTRACT Measuring cardiorespiratory variables can be challenging in developing animals, especially when they use bimodal gas exchange to maintain metabolic activity. In tadpoles, gas exchange may occur through the integument and gills when breathing in the water and through the lungs when breathing air, with varying contributions of each respiratory structure during development. The interaction between aquatic and air breathing results in a complex physiological response that may affect the cardiac cycle. Measuring the heart rate (fH) together with aquatic and aerial gas exchange in anurans during their development can be challenging, since it may involve handling small animals and/or a certain degree of invasiveness (i.e., surgery to implant electrodes). Here, we evaluated concomitantly aquatic and aerial gas exchange, lung ventilation, and fH in three stages of development of the bullfrog Lithobates catesbeianus (Shaw, 1802). We built a novel, noninvasive, closed respirometry system capable of measuring fH, aerial and aquatic gas exchange simultaneously in animals of different sizes. Our integrative analysis revealed a decrease in the heart rate and an increase in oxygen consumption during the developmental stages of the bullfrog, but there was no adjustment of heart rate after or during air breathing. Moreover, tadpoles in metamorphosis showed higher oxygen consumption in air than in water, while aquatic breathing was responsible for releasing CO2. Our results are consistent with those found in the literature, yet our study represents the first non-invasive investigation to evaluate bimodal gas exchange and heart rate simultaneously. Moreover, our setup holds potential for further advancements that would allow for controlled water and air composition. This tool could greatly facilitate the investigation of how cardiorespiratory physiology responds to varying environmental conditions.

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All known species of the Triassic archosauromorph genus Tanystropheus are known to have had the longest neck in proportion to their torso. This feature is related to a series of ventilatory challenges since an increase in neck length also increases airway length and, therefore, the volume of stagnant air that does not reach the lungs, the dead space volume. Based on this challenge, the objective of the present study was to model the type of respiratory system of Tanystropheus able to meet its metabolic demands during the early Triassic period. The modeling was based on allometric relations for morphological and physiological ventilatory and metabolic variables, and to do so, the mean body mass of Tanystropheus was estimated based on three different methods. In addition, the tracheal airflow was also estimated based on the proportions of Tanystropheus elongated neck, the results of allometric modeling, and fundamental equations of fluid mechanics. The estimation of the body mass indicated that an animal of 3.6 m would possess a body mass of 50.6 ± 21.6 kg. Allometric modeling suggested that the respiratory system best suited to Tanystropheus' oxygen demands, especially during activity, would be a generic reptilian-like respiratory system composed of multicameral lungs. The best respiratory pattern to maintain adequate tracheal flow rates and effective pulmonary ventilation would be one ventilating the relatively narrower trachea at lower frequencies to deal with tracheal dead space volume.

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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.

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The respiratory system of chelonians needs to function within a mostly solid carapace, with ventilation depending on movements of the flanks. When submerged, inspiration has to work against hydrostatic pressure. We examined breathing mechanics in Trachemys scripta while underwater. Additionally, as the respiratory system of T. scripta possesses a well-developed post-pulmonary septum (PPS), we investigated its role by analyzing the breathing mechanics of lungs with and without their PPS attached. Static compliance was significantly increased in submerged animals and in animals with and without their PPS, while removal of the PPS did not result in a significantly different static compliance. Dynamic compliance was significantly affected by changes in volume and frequency in every treatment, with submergence significantly decreasing dynamic compliance. The presence of the PPS significantly increased dynamic compliance. Submersion did not significantly alter work per ventilation, but caused minute work of breathing to be much greater at any frequency and ventilation level analyzed. Lungs with or without their PPS did not show significantly different work per ventilation when compared with the intact animal. Our results demonstrate that submersion results in significantly altered breathing mechanics, increasing minute work of breathing greatly. The PPS was shown to maintain a constant volume within the animal's body cavity, wherein the lungs can be ventilated more easily, highlighting the importance of this coelomic subdivision in the chelonian body cavity.

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Biological variables are frequently described by analyzing scaling relationships of the variable against body mass (MB). Respiratory variables are no exception and allometric relations for oxygen consumption, pulmonary ventilation, tidal volume, breathing frequency, and lung volume have been described in the literature. While the allometric relations of respiratory variables given for mammals and birds are very consistent among different studies, scaling relationships for non-avian reptiles have only been scarcely described and show considerable variation between studies. Since no comprehensive study of allometric relations of respiratory variables has been carried out comparing the different groups of non-avian reptiles, we analyzed morphological and physiological variables of the respiratory system of crocodilians, chelonians, lizards, snakes, birds, and mammals, regarding the allometric relations of each variable from a phylogenetic perspective as well as related to lung morphology. Our results indicated that few respiratory variables possess significant phylogenetic signals and that tidal volume, breathing frequency (except mammals), and air convection requirement were independent of phylogeny. Contrary to the literature, lung volume of amniotes scaled isometrically to MB, with the exception of lizards (MB0.78). Air convection requirement scaled isometrically in mammals and birds, but was more variable among non-avian reptiles, from a taxonomic perspective and in regard to different lung structures. In conclusion, respiratory variables among non-avian reptiles scaled more variably than previously expected, both according to phylogeny and to lung type, warranting future studies to explore structure-function relations of the reptilian respiratory system, especially regarding snakes and crocodilians, since these groups had very few data available for analysis.

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Carbon dioxide (CO2) and oxygen (O2) influence the breathing pattern of reptiles, especially when CO2 is in excess or O2 at low concentrations and the effects of these gases on the respiratory response varies according to the species. In addition to respiratory gases, seasonal changes can also modulate breathing pattern and ventilatory responses to hypoxia and hypercarbia. Therefore, the present study investigated the breathing pattern and ventilatory responses to hypercarbia (5% CO2) and hypoxia (5% O2) of the Neotropical lizard Tropidurus torquatus over a period of one year, covering all seasons (summer, autumn, winter and spring). Our data suggest that like other ectothermic sauropsids, Tropidurus torquatus possesses distinct ventilatory responses to hypoxia and hypercarbia, being more sensitive to changes in CO2 than in O2. Additionally, the ventilatory responses to hypoxia were more pronounced during summer and hypercanic and pos-hypercapnic ventilatory response was reduced during spring, suggesting that seasonality modulates the control of ventilation in this species.

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The "jet stream" model predicts an expired flow within the dorsal part of the buccal cavity with small air mixing during buccal pump ventilation, and has been suggested for some anuran amphibians but no other species of air breathing animal using a buccal force pump has been investigated. The presence of a two-stroke buccal pump in lungfish, i.e. expiration followed by inspiration, was described previously, but no quantitative data are available for the dead-space of their respiratory system and neither a detailed description of airflow throughout a breathing cycle. The present study aimed to assess the degree of mixing of fresh air and expired gas during the breathing cycle of Lepidosiren paradoxa and to verify the possible presence of a jet stream during expiration in this species. To do so, simultaneous measurements of buccal pressure and ventilatory airflows were carried out. Buccal and lung gases (PCO2 and PO2) were also measured. The effective ventilation was calculated and the dead space estimated using Bohr equations. The results confirmed that the two-stroke buccal pump is present in lungfish, as it is in anuran amphibians. The present approaches were coherent with a small dead space, with a very small buccal-lung PCO2 difference. In the South American lungfish the dead space (VD) as a percentage of tidal volume (VT) (VD / VT) ranged from 4.1 to 12.5%. Our data support the presence of a jet stream and indicate a small degree of air mixing in the buccal cavity. Comparisons with the literature indicate that these data are similar to previous data reported for the toad Rhinella schneideri.

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The embryonic development of parabronchi occurs mainly during the second half of incubation in precocious birds, which makes this phase sensitive to possible morphological modifications induced by O2 supply limitation. Thus, we hypothesized that hypoxia during the embryonic phase of parabronchial development induces morphological changes that remain after hatching. To test this hypothesis, chicken embryos were incubated entirely (21 days) under normoxia or partially under hypoxia (15% O2 during days 12 to 18). Lung structures, including air capillaries, blood capillaries, infundibula, atria, parabronchial lumen, bronchi, blood vessels larger than capillaries and interparabronchial tissue, in 1- and 10-day-old chicks were analyzed using light microscopy-assisted stereology. Tissue barrier and surface area of air capillaries were measured using electron microscopy-assisted stereology, allowing for calculation of the anatomical diffusion factor. Hypoxia increased the relative volumes of air and blood capillaries, structures directly involved in gas exchange, but decreased the relative volumes of atria in both groups of chicks, and the parabronchial lumen in older chicks. Accordingly, the surface area of the air capillaries and the anatomical diffusion factor were increased under hypoxic incubation. Treatment did not alter total lung volume, relative volumes of infundibula, bronchi, blood vessels larger than capillaries, interparabronchial tissue or the tissue barrier of any group. We conclude that hypoxia during the embryonic phase of parabronchial development leads to a morphological remodeling, characterized by increased volume density and respiratory surface area of structures involved in gas exchange at the expense of structures responsible for air conduction in chicks up to 10 days old.

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Many frog species reproduce in temporary ponds maintained exclusively by rainfall, thereby being exposed to drought and possibly mortality of eggs and tadpoles. Some tadpoles, however, can survive for up to 5 days out of water but few data are available regarding the effects of dehydration on their development. The aim of this study was to evaluate whether hydric stress affects the locomotor capacity and the morphology in tadpoles of two leptodactylid frog species showing different reproductive modes (Leptodactylus fuscus and Physalaemus nattereri), examining specifically: (a) difference in survival rate and body mass between tadpoles at different hydration levels, (b) the hydric stress effect on locomotor performance, (c) difference in external morphology, and (d) visceral volume among tadpoles suffering hydric stress. Tadpoles for both species were divided into two groups, one staying in 100 ml of water and the other maintained on absorbent paper with 4 ml of water for 12, 24, and 72 hr (n = 20 each). Significant differences in weight loss were found between the groups of both species, the treatment losing more weight in all stress levels. Almost half of P. nattereri tadpoles died within 36 hr of hydric stress. We found no difference in locomotor performance between groups of L. fuscus tadpoles, but significant differences in locomotor performance, tail morphometry, and visceral volume between groups of P. nattereri tadpoles. Our results suggest that hydric stress has a significant effect on locomotion and morphology of P. nattereri tadpoles but not in L. fuscus.

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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.

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Basal metabolic rate (BMR) represents the lowest level of metabolic activity capable to sustain homeostasis in an endotherm and is an important tool to compare metabolic rates of different species. Echimyidae is the most specious family within caviomorph rodents, however, little is known about the biology of its species, such as Trinomys setosus (Desmarest, 1817) and Clyomys bishopi (Ávila-Pires & Wutke, 1981), a ground and an underground dwelling echimyid, respectively. The ambient temperature and circadian effects on metabolic rate were evaluated through closed-system respirometry for these two species, as well as the circadian effects on CO2 production and respiratory exchange ratio (RER). Trinomys setosus and C. bishopi showed the lowest metabolic rates (0.56 ± 0.02 mLO2.h-1.g-1 and 0.53 ± 0.03 mLO2.h-1.g-1, respectively) at 32 °C and during the light phase. Under laboratory conditions, T. setosus showed metabolic rate variation compatible with nocturnal activity, whereas C. bishopi activity cycle remains unclear. Both species showed BMR lower than expected by allometric regressions for rodents.(AU)

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Basal metabolic rate (BMR) represents the lowest level of metabolic activity capable to sustain homeostasis in an endotherm and is an important tool to compare metabolic rates of different species. Echimyidae is the most specious family within caviomorph rodents, however, little is known about the biology of its species, such as Trinomys setosus (Desmarest, 1817) and Clyomys bishopi (Ávila-Pires & Wutke, 1981), a ground and an underground dwelling echimyid, respectively. The ambient temperature and circadian effects on metabolic rate were evaluated through closed-system respirometry for these two species, as well as the circadian effects on CO2 production and respiratory exchange ratio (RER). Trinomys setosus and C. bishopi showed the lowest metabolic rates (0.56 ± 0.02 mLO2.h-1.g-1 and 0.53 ± 0.03 mLO2.h-1.g-1, respectively) at 32 °C and during the light phase. Under laboratory conditions, T. setosus showed metabolic rate variation compatible with nocturnal activity, whereas C. bishopi activity cycle remains unclear. Both species showed BMR lower than expected by allometric regressions for rodents.

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The South American lungfish Lepidosiren paradoxa is an obligatory air-breathing fish possessing well-developed bilateral lungs, and undergoing seasonal changes in its habitat, including temperature changes. In the present study we aimed to evaluate gas exchange and pulmonary breathing pattern in L. paradoxa at different temperatures (25 and 30°C) and different inspired O2 levels (21, 12, 10, and 7%). Normoxic breathing pattern consisted of isolated ventilatory cycles composed of an expiration followed by 2.4±0.2 buccal inspirations. Both expiratory and inspiratory tidal volumes reached a maximum of about 35mlkg-1, indicating that L. paradoxa is able to exchange nearly all of its lung air in a single ventilatory cycle. At both temperatures, hypoxia caused a significant increase in pulmonary ventilation (V̇E), mainly due to an increase in respiratory frequency. Durations of the ventilatory cycle and expiratory and inspiratory tidal volumes were not significantly affected by hypoxia. Expiratory time (but not inspiratory) was significantly shorter at 30°C and at all O2 levels. While a small change in oxygen consumption (V̇O2) could be noticed, the carbon dioxide release (V̇CO2, P=0.0003) and air convection requirement (V̇E/V̇O2, P=0.0001) were significantly affected by hypoxia (7% O2) at both temperatures, when compared to normoxia, and pulmonary diffusion capacity increased about four-fold due to hypoxic exposure. These data highlight important features of the respiratory system of L. paradoxa, capable of matching O2 demand and supply under different environmental change, as well as help to understand the evolution of air breathing in lungfish.

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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.

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