Aquatic Respiration: An Unusual Strategy in the Hellbender Cryptobranchus alleganiensis alleganiensis (Daudin).

Separate and simultaneous determinations of aerial and aquatic gas exchange in the giant salamander, Cryptobranchus alleganiensis alleganiensis (Daudin) were made at 5 degrees , 15 degrees , and 25 degrees C. The aquatic respiration of this animal accounts for over 90 percent of the total volume of oxygen consumed and 97 percent of the total volume of carbon dioxide released at all temperatures. The lungs of these individuals are large transparent sacs which are poor respiratory organs; the lungs probably function more as hydrostatic structures than as gas exchangers. This animal is the largest aquatic vertebrate that lacks gills and yet utilizes almost exclusively an aquatic mode of respiration. Specialized cutaneous modifications, a unique body form, and a peculiar behavioral mechanism are of considerable adaptative significance, and confer to the skin the effectiveness of a veritable "gill."

Thermoregulation in a brooding female Indian python, Python molurus bivittatus.

At varying environmental temperatures, measurements of body temperatures and gas exchange of a female Indian python (Python molurus bivittatus) show that during the brooding period this animal can regulate its body temperature by physiological means analogous to those in endotherms. Ambient temperatures below 33 degrees C result in spasmodic contractions of the body musculature with a consequent increase in metabolism and body temperature.

Photoperiod acclimation and 24-hour variations in the critical thermal maxima of a tropical and a temperate frog.

The effect of photoperiod on the upper thermal tolerance of two species of frogs was studied by using the critical thermal maximum (CTM) as the end point. Both species are heliotropic and from temperate climates, but Hyla labialis lives under a near constant tropical photoperiod while Rana pipiens lives under a varying temperatezone photoperiod. The CTM of both species was studied over a 24-hour period to determine if a rhythm of temperature tolerance exists. In all but one of the acclimatization conditions used, the CTM of R. pipiens was higher than that of H. labialis. This agrees with what is known of their thermal ecology. Photoperiod significantly affects the CTM of both species. For Rana pipiens long (LD 16:8) photoperiods result in significantly higher thermal tolerance than short (LD 8:16) or moderate (LD 12:12) photoperiods at both 15 and 25° C. H. labialis shows a different pattern, having highest CTM at 25°C, LD 12:12 and lowest at 15°C, LD 12:12. When acclimated to a short (LD 8:16) photoperiod certain aspects of the frogs' tolerance of high temperatures are altered. At the same acclimatization the CTM of R. pipiens is higher than that of H. labialis, except under a combination short light regime and low temperature, and H. labialis at LD 8:16 shows no thermal acclimation between 15 and 25°C. Significant variation in the CTM over a 24-hour period occurred in H. labialis acclimatized at 25°C, LD 12:12 and R. pipiens at 25°C, LD 8:16 and 15°C, LD 12:12. For both species the 24-hour rhythm of temperature tolerance, when it occurs at LD 12:12, might be of adaptive value. Times of highest thermal tolerance are in the late morning or early afternoon and lowest tolerance is during the dark period. For R. pipiens under the unnatural combination of 25°C, LD 8:16, the pattern is reversed. When all three significant cycles are phase shifted so that the times of highest tolerance coincide, the pattern of the curves is very similar.

Histamine and histamine receptors: behavioral thermoregulation in the salamander Necturus maculosus.

Low doses (0.01, 0.1 mg/kg, i.p.) of histamine (HA) caused selection of significantly lower temperatures, and higher doses (0.5, 1.0 mg/kg) increased temperatures by mudpuppies in linear thermal gradients. Injection of the HA precursor, L-histidine (500 mg/kg) produced an increase in the temperatures selected. Results from injections of HA H1-receptor agonist (2-pyridylethylamine) and antagonist (pyrilamine), and H2-receptor agonist (dimaprit) and antagonist (cimetidine) had significant effects on thermoregulation; H1-receptors may mediate behavioral hyperthermia and H2-receptors behavioral hypothermia. Responses to these histaminic compounds are significantly influenced by the time of day at which the responses are measured and by season and acclimation temperature. The equivalent behavioral responses in both endotherms and ectotherms to agents which produce physiological hyperthermia and hypothermia are probably behavioral hypothermia ("cold seeking") and behavioral hyperthermia ("heat seeking"), respectively.

Erythrocytic organic phosphates: diel and seasonal cycles in the frog, Rana berlandieri.

Hemoglobin (Hb) concentration, hematocrit (Hct), total organic phosphates (Ptot), nucleoside triphosphates (NTP), 2,3-diphosphoglycerate (DPG), and "inositol polyphosphates" (IPP) were measured in the erythrocytes of adult frogs at different times of day in winter and summer after acclimatization to 15 degrees C and an LD 12:12 photoperiod. The same measurements were also made on animals acclimated to LD 16:8 in summer. All of the measured parameters varied significantly at different times of the day and between seasons in animals acclimated to an LD 12:12 photoperiod. Summer animals acclimated to LD 16:8 showed significant diel cycles only in Hct and Hb.

Decreased thermal tolerance in Mus musculus with melatonin and chlorpromazine.

Adult Mus musculus, previously acclimatized for two weeks to 25 degrees C and an LD 12:12 photoperiod, were injected (i.p.) with daily doses of melatonin (4.0 mg) or chlorpromazine (20.0 mg kg-1) for 2 days. Both drugs significantly reduced the critical thermal maximum (CTM) and the elevated defended temperature (EDT). Melatonin significantly increased the length of EDT but chlorpromazine had no effect on EDT. Both drugs reduce thermal tolerance in ectotherms, but this is apparently the first demonstration that melatonin and chlorpromazine increase sensitivity to high temperature in an endotherm.

Influence of temperature and photoperiod on plasma melatonin in the mudpuppy, Necturus maculosus.

A melatonin (MEL) radioimmunoassay employed previously only in mammals was used to estimate plasma MEL in a salamander, Necturus maculosus. Validation procedures included thin-layer chromatography of plasma extracts, parallel inhibition curves of authentic MEL and serially diluted plasma and plasma extracts and quantitative recovery of authentic MEL added to pooled Necturus plasma. A diel cycle of plasma MEL was demonstrated in mudpuppies acclimatized for a minimum of 3 weeks under a 12L:12D photoperiod and 15 +/- 1 degrees and sampled in late March. The MEL cycle persisted under a reversed photoperiod (lights on, 1800 hr), but the amplitude of the MEL peak was diminished, and the peak was more than 180 degrees out of phase with the corresponding peak under a normal photoperiod. In animals acclimated to 5 degrees in mid-June under a 12L:12D photoperiod, the diel cycle of plasma MEL continued, but both midphotophase and midscotophase concentrations were reduced compared with 15 degrees controls. The diel cycle was also present in animals acclimated to 25 degrees, but the decrease in MEL concentrations was less marked than that in 5 degrees animals. Photoperiod is apparently the primary cue for cycles in plasma MEL in Necturus, but the cycle can be influenced by temperature.

Parietalectomy and thermal selection in the lizard Sceloporus magister.

Lizards were acclimatized to various regimes of temperature (15 degrees, 35 degrees C) and photoperiod (LD 08:16, 16:8). Parietalectomized, sham operated, and control animals were placed in thermal gradients and their body temperatures monitored for periods of up to 15 days. Daily rhythms of thermal selection were evident with higher temperatures selected in late photophase and lower temperatures selected in scotophase. Thermal selection was more variable in scotophase than in photophase. Parietalectomized lizards chose significantly higher body temperatures than did shams or controls. Acclimatization to temperature and photoperiod had little or no effect on the thermal preferendum. The preferred body temperature of controls was 33.2 degrees C +/- 3.59. Length of time in the gradient tended to change thermal preference, especially in lizards acclimated to 15 degrees C. Through interactions with the pineal gland the parietal eye is probably important in synchronizing many bodily functions with photoperiod.

Aerobic and anaerobic contributions to sustained activity in Xenopus laevis.

In conditions of declining water PO2, Xenopus obtains the majority of resting oxygen needs from lung breathing at 15 degrees and 25 degrees C. The critical oxygen tension was 120 +/- 9 mm Hg at 15 degrees C, and 90 +/- 10 mm Hg at 25 degrees C. During 30 min stimulation of activity to complete exhaustion at 15 degrees C, frogs exhibited an aerobic capacity of 1.7 microliter O2.g-1.h-1 and accumulated 2.22 mg lactate . g-1. Following activity these animals exhibited an oxygen debt of 49.2 microliter O2.g-1. At 25 degrees C, Xenopus had an aerobic capacity of 16.1 microliter O2.g-1 and accumulated 1.94 mg lactate . g-1. Following activity, 25 degrees C frogs exhibited an oxygen debt of 261.9 microliter O2.g-1. Thus at 15 degrees C, Xenopus acquires 85% of the ATP used during a bout of sustained exhaustive activity from anaerobic sources, and 98% at 25 degrees C. Recovery from exhaustive activity was complete after 5 h at 25 degrees C and 9 h at 15 degrees C. Comparison of these data with those of other amphibians stimulated to sustained activity suggests that aquatic amphibians can tolerate a larger lactate load than can terrestrial forms.

Blood changes in Bufo cognatus following acute heat stress.

1. Various blood constituents were measured in an attempt to identify the effects of exposure of Great Plains toads to the critical thermal maximum (CTmax) and determine the time course of the onset of and recovery from these effects. 2. Tests for generalized tissue damage including serum glutamic-oxalacetic and glutamic-pyruvic transaminases (SGOT, SGPT), total protein and blood urea nitrogen (BUN) were unaffected by acute thermal stress. 3. Hematocrit, erythrocyte number, mean cell volume and hemoglobin concentration were also unchanged. 4. Blood glucose, lactic acid and creatine phosphokinase (CPK) levels all increased significantly. 5. Blood pH, PO2 and [HCO3-] also increased with acute heat stress while PCO2 decreased. 6. Long-term exposure to temperatures near the CTmax may cause severe tissue damage. Acute thermal stress does not appear to cause damage other than the short-term, reversible effects of strong physical exercise.

Oxygen transfer characteristics of the blood of reedfish, Erpetoichthys calabaricus.

Oxygen transport characteristics and phosphate compounds were measured in the blood of reedfish, Erpetoichthys calabaricus, a bimodal breather. Blood from reedfish possessed the following values (mean +/- SD): hematocrit (21.7 +/- 0.4%), hemoglobin concentration (7.53 +/- 1.75 g%), red blood cell count (0.45 +/- 0.10 X 10(6)/mm3) and oxygen capacity (10.1 +/- 2.3 vol%). Although hematocrit, hemoglobin concentration, red blood cell count and oxygen capacity were all highly intercorrelated (P less than 0.01 in all cases), none of these parameters were significantly correlated with sex, weight or length in our sixteen fish sample. Erythrocyte volumes equalled 480 micrometers3, showed less variation (CV = 10.4%) and did not correlate with any other measured variable. Blood oxygen dissociation curves were sigmoidal and the P50's equalled 17.34 +/- 3.04 at 1% CO2 and 25 degrees C. Mean Bohr shift (delta log P50/delta pH) was -0.274 +/- 0.087. Temperature strongly influenced blood oxygen affinity. At 1% CO2, delta log P50/delta T equalled 0.026 +/- 0.006 (mean +/- SD). These hematological properties indicate that the blood of reedfish is similar to those of other tropical air-breathering species. Concentrations of total phosphate in the erythrocytes and percentage of total phosphate bound as nucleotide triphosphates were high. Surprisingly, 2,3diphosphoglycerate was found which has been reported in the erythrocytes of only two other fish species. Blood characteristics of reedfish exposed to air for 4 hr with one exception (Hill numbers) were not significantly different from water exposed controls. This suggests that the reedfish does not possess blood respiratory mechanisms to facilitate respiration solely by air-breathing.

Aquatic life at high altitude: respiratory adaptations in the Lake Titicaca frog, Telmatobius culeus.

Telmatobius culeus has a combination of behavioral, morphological and physiological adaptations which allows an aquatic life in cool (10 degrees C) O2-saturated (at 100 mm Hg) waters at high altitude (3812 m). The skin surgace area is increased by pronounced folds and the cutaneous capillaries penetrate to the outer layers of the skin. The erythrocyte volume (394 mu3) is the smallest reported for amphibians. The P50 (15.6 at ph 7.65 and 10 degrees C) is the lowest, and the erythrocyte count (729 - 103/mm3) the highest for an anuran. The O2 capacity (11.7 vol%), hemoglobin (8.1 g/100 ml), hemoglobin concentration (0.281 pg/mu3) and hematocrit (27.9%) measured at 18 degrees C and 3800 m are all elevated in comparison with most amphibians. The O2 dissociation curve is sigmoid (n = 2), the Bohr factor is small (deltalog P50/deltapH = -0.30) and the buffering capacity (-8.9 m M HCO3 - 1-1) is typical for an aquatic amphibian. The metabolic rate (14.1 mul -g-1-h-u) is the lowest reported for a frog and among amphibians only the giant salamanders (Amphiuma, Necturus and Siren) have lower values. If prevent from surfacing in hypoxic waters, the frogs ventilate the skin by "bobbing" behavior; if allowed to surface, they will ventilate the small lungs and the metabolic rate increases to 23 mul-g-1-h-1.

Alarm response by a plethodontid salamander (Desmognathus ochrophaeus): Conspecific and heterospecific "Schreckstoff".

The detection of chemical alarm cues plays an important role for predator avoidance in many taxonomic groups, but little is known about the presence of such chemical cues in adult or caudate amphibians. We investigated the response (i.e., aversion or nonaversion) to chemical cues from damaged salamander skin and mealworms (Tenebrio molitor) in the plethodontid salamander,Desmognathus ochrophaeus. Avoidance responses were demonstrated to skin extracts of both conspecific and heterospecific salamanders. However, salamanders (D. ochrophaeus) did not avoid heated conspecific skin, fresh conspecific viscera, fresh mealworm, or freshPlethodon richmondi skin extracts. These results indicate that chemical alarm cues are: (1) present in the skin ofDesmognathus salamanders, (2) not present in mealworm or the viscera ofDesmognathus salamanders, and (3) denatured or deactivated by heating. These results also suggest that an avoidance response to chemical cues from damaged conspecifics has adaptive value in predator avoidance in terrestrial as well as aquatic vertebrates.