Comparison of stress induced by manual restraint and immobilisation in the estuarine crocodile, Crocodylus porosus.

This study compared the stress induced in captive estuarine crocodiles, Crocodylus porosus, by two different handling methods: manual restraint (noosing with ropes) and immobilization by electro-stunning. To stun, a short charge (approx. 6 s) at 110 V was delivered to the back of the necks of C. porosus using a custom-built device, which immobilized the animals for 5-10 min. Immobilized and restrained animals were measured and sexed, and the condition of the skin assessed. Blood samples were taken from some animals immediately after restraint or immobilization. Other animals were returned to their pens to recover for periods of 30 min, 1, 4, 12, 24 or 48 hours after which they were stunned and blood samples taken. Individual animals (mean body length 1.96 m, N=99) were bled only once. Haematocrit and haemoglobin concentrations were measured and plasma samples were analysed for corticosterone, glucose and lactate levels. Following restraint, there were significant increases in haematocrit, haemoglobin, glucose, lactate and corticosterone concentrations in C. porosus. For restrained animals, recovery to baseline levels occurred after approximately 8 hours. The stress response of stunned animals was significantly reduced compared to manually captured and restrained crocodiles. Both groups showed a significant increase in haematocrit, haemoglobin concentration and lactate levels, however the magnitude of change was significantly reduced, and recovery was faster in stunned animals. No increase in either glucose or corticosterone levels occurred with immobilisation. The results imply that immobilization by electro-stunning is much less stressful.

Changes in heart rate are important for thermoregulation in the varanid lizard Varanus varius.

Laboratory studies and a single field study have shown that heart rate in some reptiles is faster during heating than during cooling at any given body temperature. This phenomenon, which has been shown to reflect changes in peripheral blood flow, is shown here to occur in the lizard Varanus varius (lace monitor) in the wild. On a typical clear day, lizards emerged from their shelters in the morning to warm in the sun. Following this, animals were active, moving until they again entered a shelter in the evening. During their period of activity, body temperature was 34-36 degrees C in all six study animals (4.0-5.6 kg), but the animals rarely shuttled between sun and shade exposure. Heart rate during the morning heating period was significantly faster than during the evening cooling period. However, the ratio of heating to cooling heart rate decreased with increasing body temperature, being close to 2 at body temperatures of 22-24 degrees C and decreasing to 1.2-1.3 at body temperatures of 34-36 degrees C. There was a significant decrease in thermal time constants with increasing heart rate during heating and cooling confirming that changes in heart rate are linked to rates of heat exchange.

Electrical activation of the ventricular myocardium of the crocodile Crocodylus johnstoni: a combined microscopic and electrophysiological study.

We mapped the sequence of ventricular depolarization in the crocodile Crocodylus johnstoni. We also attempted to find specialized conduction tissue within the ventricular myocardium. Electrical recordings with miniature multi-point electrodes revealed two strands of rapidly conducting tissue (channels) within the interventricular septum, suggestive of conductive tissue pathways. From these septal channels, wavefronts of excitation swept around each ventricle. Electrical recordings did not indicate that there was conductive tissue in the wall of either ventricle. Similarly, microscopic studies of the septal channels provided no indication of specialized conductive tissue. We suggest that the channels of early septal depolarization provide the crocodile heart with a high speed depolarization pathway functionally analogous to a rudimentary conductive system.

Field test of a paradigm: hysteresis of heart rate in thermoregulation by a free-ranging lizard (Pogona barbata).

The discovery that changes in heart rate and blood flow allow some reptiles to heat faster than they cool has become a central paradigm in our understanding of reptilian thermoregulation. However, this hysteresis in heart rate has been demonstrated only in simplistic laboratory heating and cooling trials, leaving its functional significance in free-ranging animals unproven. To test the validity of this paradigm, we measured heart rate and body temperature (Tb) in undisturbed, free-ranging bearded dragons (Pogona barbata), the species in which this phenomenon was first described. Our field data confirmed the paradigm and we found that heart rate during heating usually exceeded heart rate during cooling at any Tb. Importantly, however, we discovered that heart rate was proportionally faster in cool lizards whose Tb was still well below the 'preferred Tb range' compared to lizards whose Tb was already close to it. Similarly, heart rate during cooling was proportionally slower the warmer the lizard and the greater its cooling potential compared to lizards whose Tb was already near minimum operative temperature. Further, we predicted that, if heart rate hysteresis has functional significance, a 'reverse hysteresis' pattern should be observable when lizards risked overheating. This was indeed the case and, during heating on those occasions when Tb reached very high levels (> 40 degrees C), heart rate was significantly lower than heart rate during the immediately following cooling phase. These results demonstrate that physiological control of thermoregulation in reptiles is more complex than has been previously recognized.

Body temperatures and activity patterns of Tasmanian devils (Sarcophilus harrisii) and eastern quolls (Dasyurus viverrinus) through a subalpine winter.

During a field study of carnivorous dasyurid marsupials in subalpine Tasmania, the trapping success for Tasmanian devils (Sarcophilus harrisii), but not for spotted-tailed quolls (Dasyurus maculatus) or eastern quolls (Dasyurus viverrinus), was significantly lower when winter weather conditions turned to sleet or snow or when deep snow lay on the ground. This field study was instigated to determine if devils and eastern quolls spend more time in burrows in severe weather conditions and if they enter torpor. Torpor is known to occur in eutherian mammals as large as devils and in a similar-sized congeneric marsupial, the western quoll (Dasyurus geoffroyi). Using radiotelemetry, body temperatures of Tasmanian devils and eastern quolls ranging freely in their natural habitat were monitored throughout winter. Neither species was observed in torpor, even under prolonged severe weather conditions, and the number of hours spent active not did differ between summer and winter or between moderate and severe winter weather conditions. Body temperatures averaged 36.5 degrees C (SD = 0.079, range of 33.5 degrees-38.6 degrees C) for the three male eastern quolls and 35.7 degrees C (SD = 0.575, range of 31.3 degrees-37.5 degrees C) for the four (male and female) devils. A diel cycle in body temperature occurred in both species; temperatures rose each evening when animals became active, remained high throughout the night despite ambient temperatures falling to the diel minima, and were lower during the day when the individuals were inactive in dens. The amplitude of this cycle was greater in eastern quolls (1.1 degrees C, SD = 0.142) than in devils (0.6 degree C, SD = 0.252).

Neuropeptide immunoreactivity and co-existence in cardiovascular nerves and autonomic ganglia of the estuarine crocodile, Crocodylus porosus, and cardiovascular effects of neuropeptides.

The two aortas of the crocodile are in open connection at two sites, the foramen of Panizzae immediately outside the ventricles, and the arterial anastomosis at the level of the gut. The present study was performed to elucidate the innervation of the cardiovascular structures of the crocodile, in part to provide a further basis for the assumption that the apertures of the foramen and the anastomosis may be altered, possibly leading to changes in the flow profiles of the central vessels. The presence of smooth muscle arranged at the circumference of the foramen and in the walls of the anastomosis was demonstrated. The cardiovascular structures were innervated by nerves containing co-existing tyrosine hydroxylase, NPY and somatostatin immunoreactivities, which also occurred in neurons of the sympathetic ganglia. CGRP and substance P immunoreactive material co-existed in cardiovascular nerves, and in the nodose ganglion. In addition, bombesin, VIP and galanin immunoreactive nerves were found. Effects of neuropeptides on blood flows and blood pressures were studied in vivo. Substance P increased all blood flows measured, NPY increased the flow through the arterial anastomosis while neurotensin caused an initial decrease in the flow through the arterial anastomosis. In conclusion, there is a rich innervation of the heart and major vessels of the estuarine crocodile, including the foramen of Panizza and the arterial anastomosis. These nerves possibly regulate the distribution of blood in the cardiovascular system, which is further suggested by the results of the injection of neuropeptides.

Ventilatory and metabolic responses to hypoxia in the echidna, Tachyglossus aculeatus.

Oxygen consumption (VO2), CO2 production (VCO2), and minute ventilation (VE) together with breathing pattern were measured in echidnas during normoxia and hypoxia. In normoxia, VO2, VCO2, and VE were all found to be approximately 30% of the allometric prediction for a eutherian. As a consequence VE/VO2 and VE/VCO2 are as predicted for a mammal. This is in contrast to previous reports on the echidna in which the VE was shown to be low and the echidna, subsequently, to be in a state of hypoventilation. It is possible that the difference between this and previous studies is related to the resting state of the echidna; echidnas in this study adopted a curled-up "sleeping" posture, and measurements were made without tactile disturbance. Breathing pattern was typical of a semifossorial species in that inspiration time to total breath time was short when compared with the normal eutherian value. In graded hypoxia VE increased [threshold fractional concentration of inspired O2 (FIO2) = 0.125], predominantly the result of changes in frequency achieved through a shortening in expiration time. In acute hypoxia (FIO2 = 0.10) VE/metabolic rate showed a tendency to increase, mainly because of the increase in VE. Approximately 50% of the increase in VE could be attributed to the 25% increase in VO2 and VCO2 that occurred in acute hypoxia. Given that the general mammalian response to hypoxia is a drop in metabolic rate, possible reasons as to why the echidna does not decrease metabolic rate in hypoxia are discussed.

Blood pressure and heart rate during tonic immobility in the black tipped reef shark,Carcharhinus melanoptera.

Tonic immobility was induced in black tipped reef sharks (Carcharhinus melanoptera) and heart rate and ventral aortic blood pressure recorded. Without branchial irrigation, tonic immobility was correlated with a significant depression in blood pressure and heart rate irrespective of the sharks being in air or in water. Tonic immobility with branchial irrigation resulted in a significant increase in blood pressure in sharks in air, but not in water. Heart rate was unchanged when the gills were irrigated. Intra-arterial injections of atropine abolished the bradycardia and blood pressure rise associated with tonic immobility. We conclude that, during tonic immobility, sharks are able to receive afferent information from the ventilatory system and make appropriate responses via the vagus nerve.

Redistribution of air within the lungs may potentiate "fright" bradycardia in submerged crocodiles (Crocodylus porosus).

1. Voluntary undisturbed dives by Crocodylus porosus were short in duration (3.08 +/- 1.87 min, mean +/- SD) and accompanied by a small but significant bradycardia (14.3 +/- 5.9% drop). 2. When crocodiles were disturbed underwater there was a rapid onset of "fright" bradycardia, to 65 +/- 6.0% of surface heart rates and dive durations were prolonged to 19.6 +/- 1.8 min. 3. The development of "fright" bradycardia was not accompanied by any increase in intratracheal pressure or expulsion of lung gas. However, sustained contraction of the abdomen and expansion of the thorax revealed a redistribution of air anteriorly within the lungs. 4. We propose that the redistribution of air within the lungs may generate an afferent signal which potentiates the initiation of a severe, dive-prolonging bradycardia.

Hibernation in a monotreme, the echidna (Tachyglossus aculeatus).

The body temperatures of five echidnas in Australia's Southern Alps were monitored by radio telemetry from February to December 1987. All five hibernated throughout the winter, showing very low body temperatures (4-9 degrees C, close to ambient) when torpid, compared with 28-33 degrees C in a typical day during the active season. Spontaneous arousals from hibernation occurred every 2-3 weeks, during which body temperatures rose rapidly to over 30 degrees C for several hours before dropping to be close to ambient again. The identification of "classical" hibernation in a monotreme, with a similar pattern to that seen in Eutheria and in an animal as large as the largest eutherian hibernator, has important implications for current ideas about the evolution of endothermy.

A simple and inexpensive tonometry system for use with microlitre blood samples.

The construction of a blood tonometering system based on the vibrating action of a firebell is described. Its performance has been assessed with human blood at 37 degrees C and in use with Antarctic fish blood (Pagothenia borchgrevinki) at -1.5 degrees C.

Osmoregulation of the cane toad, Bufo marinus, in salt water.

Adult cane toads, B. marinus, survived in salinities up to 40% sea-water (SW). Pre-exposure to 30, then 40% SW, increased the survival time of toads in 50% SW. Plasma from toads acclimated to salt water is hyperosmotic to the environment--a result of increased plasma sodium, chloride and urea concentrations. When toads were placed in tap-water and 20% SW, all significant changes to plasma sodium, chloride, urea and osmotic pressure occurred within the first 2 days of exposure. When toads were placed in 30 and 40% SW environments, the increases in plasma sodium and chloride concentrations occurred within the first 2 days of exposure while urea and total osmotic pressure continued to rise until some time between 2 and 7 days exposure.

Salt Glands in the Tongue of the Estuarine Crocodile Crocodylus porosus.

The apparent absence of salt glands in marine and estuarine Crocodilia has long been a puzzle. However, we have identified glands in the tongue of Crocodylus porosus which exude a concentrated secretion of sodium chloride. The glands are similar in ultrastructure to other reptilian salt glands and undoubtedly play a major role in electrolyte regulation.

Respiratory properties of the blood of Crocodylus porosus.

The blood of Crocodylus porosus has a high oxygen capacity (5.5 mmol . 1-1 at hematocrit = 28%). The shape of the oxygen equilibrium curve of the blood is described by 'n' = 2.7 in the physiological range of PCO2, and its oxygen affinity is described by the equation: log10P50 = 0.4163 + 0.0200 T degrees C + 0.3763 log10PCO2. Thus, the blood has a low oxygen affinity which is strongly sensitive to both temperature and PCO2. There is a high buffering capacity, 37 mmol . (1 . pH)-1, and a large Haldane effect, 0.93 mmol CO2 (mmol Hb)-1. The fixed-acid Bohr effect seems to be much reduced in comparison to the CO2-specific Bohr effect. We discuss the possibility that low levels of red cell organic phosphate may be an adaptive strategy to desensitise P50 to changes in plasma pH. The significance of the blood respiratory properties is discussed in terms of the life style of C. porosus, particularly in relation to ectothermy and diving.

Methemoglobin reduction in crocodile blood: are high levels of MetHb typical of healthy reptiles?

In 82 wild-caught Crocodylus porosus, levels of NADH-MetHb reductase and GSH seem adequate to maintain hemoglobin in its reduced functional state. Studies of C. porosus erythrocytes in vitro show reduction of metHb in the presence of lactate, glucose and plasma, but not pyruvate. These findings, together with recent data which show low metHb in a variety of reptiles, cast doubt on the accepted view that high levels of MetHb are typical of healthy reptiles. One explanation for the sharp contrast between earlier and more recent data could be technical. We found low metHb in Crocodylus johnstoni, Chelodina longicollis and Sphenomorphus quoyi. However, high and variable vales reminiscent of many of the earlier data were obtained by omitting final centrifugation prior to spectrophotometry. Interestingly, this step is not part of the standard clinical method but is necessary in analyses of blood with nucleated red cells. These observations suggest that high metHb may not be typical of reptiles after all.

Survival and growth of hatchling Crocodylus porosus in saltwater without access to fresh drinking water.

It has been suggested that C. porosus select nest sites which provide a source of freshwater for hatchlings during the dry season. From a mark-recapture study, we conclude that hatchling C. porosus can survive and grow in hyperosmotic saltwater without drinking freshwater. Hence, the siting of nests is unlikely to be the consequence of a requirement by hatchlings for freshwater. Considered along with other information, our observations imply that hatchling C. porosus have functional salt glands.