Biomedical subjects

A F Bennett

Publications and source records attributed to A F Bennett.

At least 19 recordsLinked to original sources

PubMed2006

Recovery from an activity-induced metabolic acidosis in the American alligator, Alligator mississippiensis.

The metabolic acidosis resulting from an intense exercise bout is large in crocodilians. Here we studied recovery from this pH perturbation in the American alligator. Metabolic rate, minute ventilation, arterial pH and gases, and strong ion concentration were measured for 10 h after exhaustion to elucidate the mechanisms and time course of recovery. Exhaustion resulted in a significant increase in lactate, metabolic rate, and ventilation, and a decrease in arterial PCO2), pH and bicarbonate. By 15 min after exhaustion, oxygen consumption returned to rest though carbon dioxide excretion remained elevated for 30 min. Arterial PO2), [Na+], and [K+], increased following exhaustion and recovered by 30 min post-exercise. Minute ventilation, tidal volume, [Cl-], and respiratory exchange ratio returned to resting values by 1 h. The air convection requirement for oxygen was elevated between 15 and 60 min of recovery. Breathing frequency and pH returned to resting values by 2 h of recovery. Lactate levels remained elevated until 6 h post-exercise. Arterial PCO2) and [HCO3-] were depressed until 8 h post-exercise. Compensation during recovery of acid-base balance was achieved by altering ventilation: following the initial metabolic acidosis and titration of bicarbonate, a relative hyperventilation prevented a further decrease in pH.

Acidosis↗

PubMed2006

Metabolic and blood gas dependence on digestive state in the Savannah monitor lizard Varanus exanthematicus: an assessment of the alkaline tide.

A large alkaline tide (up to 20 mmol l(-1) increase in bicarbonate concentration [HCO3-] with an accompanied increase in blood pH) has previously been reported for some carnivorous reptiles within 24 h after ingesting a large meal. This phenomenon has been attributed to the secretion of large amounts of H+ ions into the stomach, which is required for digestion of large prey items. To test the generality of this phenomenon in carnivorous reptiles, this study quantified the metabolic and acid-base status of the Savannah monitor lizard, Varanus exanthematicus, during digestion at 35 degrees C. Following a meal of approximately 10% of body mass, V(O2) and V(CO2) were measured continuously and arterial pH, blood gases and strong ions were measured every 8 h for 5 days. During peak digestion (24 h post feeding), V(O2) and V(CO2) increased to approximately threefold fasting values (V(O2), 0.95-2.57 ml min(-1) kg(-1); V(CO2) 0.53-1.63 ml min(-1) kg(-1)) while respiratory exchange ratio (R) remained constant (0.62-0.73). During digestion, arterial P(CO2) increased (from 4.6 kPa to 5.8 kPa), and [HCO3-] also increased (from 24.1 mmol l(-1) to 40.3 mmol l(-1)). In contrast to early studies on crocodilians, arterial pH in V. exanthematicus remained relatively stable during digestion (7.43-7.56). Strong ions contributed little to the acid-base compensation during the alkalosis. Collectively the data indicate that the metabolic alkalosis associated with H+ secretion (as indicated by increased plasma bicarbonate) is partially compensated by a respiratory acidosis.

Animal Feed↗

PubMed2006

The energetic consequences of dietary specialization in populations of the garter snake, Thamnophis elegans.

We investigated the intraspecific variation in digestive energetics between dietary specialist and generalist populations of the Western Terrestrial garter snake (Thamnophis elegans) in northern California. Coastal populations have a specialized diet of slugs and inland populations have a generalized diet of fish, anurans, mice and leeches. The difference in prey preference between the two populations is congenital, heritable and ontogenetically stable. To examine energetic specializations and trade-offs in these populations, we measured the net assimilation efficiency of each snake population on both slug (Ariolimax columbianus) and fish (Rhinichthys osculus) diets. The net assimilation efficiency was measured during digestion of a meal and continued until metabolic rate re-attained prefeeding levels. Coastal snakes were able to utilize 62% more of the ingested energy towards production from slug diets through both increased assimilation of nutrients and reduced digestive costs. For fish, assimilation and digestive costs were the same in both coastal and inland populations. These results support the hypothesis that snakes with specialized diets can evolve physiological traits to extract nutrients more efficiently. However, there was no apparent trade-off on the more generalized diet that was associated with this specialization.

Adaptation, Physiological↗

PubMed2005

The effect of meal composition on specific dynamic action in burmese pythons (Python molurus).

We quantified the specific dynamic action (SDA) resulting from the ingestion of various meal types in Burmese pythons (Python molurus) at 30 degrees C. Each snake was fed a series of experimental meals consisting of amino acid mixtures, simple proteins, simple or complex carbohydrates, or lipids as well as meals of whole animal tissue (chicken breast, beef suet, and mouse). Rates of oxygen consumption were measured for approximately 4 d after feeding, and the increment above standard metabolic rate was determined and compared to energy content of the meals. While food type (protein, carbohydrate, and lipid) had a general influence, SDA was highly dependent on meal composition (i.e., amino acid composition and carbohydrate structure). For chicken breast and simple carbohydrates, the SDA coefficient was approximately one-third the energetic content of the meal. Lard, suet, cellulose, and starch were not digested and did not produce measurable SDA. We conclude that the cost of de novo protein synthesis is an important component of SDA after ingestion of protein meals because (1) simple proteins, such as gelatin and collagen, did not stimulate levels of SDA attained after consumption of complete protein, (2) incomplete mixtures of amino acids failed to elicit the SDA of a complete mixture, and (3) the inhibition of de novo protein synthesis with the drug cycloheximide caused a more than 70% decrease in SDA. Stomach distension and mechanical digestion of intact prey did not cause measurable SDA.

Amino Acids↗

PubMed2001

Energetics of side chain packing in staphylococcal nuclease assessed by exchange of valines, isoleucines, and leucines.

To examine the importance of side chain packing to protein stability, each of the 11 leucines in staphylococcal nuclease was substituted with isoleucine and valine. The nine valines were substituted with leucine and isoleucine, while the five isoleucines, previously substituted with valine, were substituted with leucine and methionine. These substitutions conserve the hydrophobic character of these side chains but alter side chain geometry and, in some cases, size. In addition, eight threonine residues, previously substituted with valine, were substituted with isoleucine to test the importance of packing at sites normally not occupied by a hydrophobic residue. The stabilities of these 58 mutant proteins were measured by guanidine hydrochloride denaturation. To the best of our knowledge, this is the largest library of single packing mutants yet characterized. As expected, repacking stability effects are tied to the degree of side chain burial. The average energetic cost of moving a single buried methyl group was 0.9 kcal/mol, albeit with a standard deviation of 0.8 kcal/mol. This average is actually slightly greater than the value of 0.7-0.8 kcal/mol estimated for the hydrophobic transfer energy of a methylene from octanol to water. These results appear to indicate that van der Waals interactions gained from optimal packing are at least as important in stabilizing the native state of proteins as hydrophobic transfer effects.

Amino Acid Substitution↗

PubMed2001

Evolutionary adaptation to temperature. IX. Preadaptation to novel stressful environments of Escherichia coli adapted to high temperature.

Stressful environments may be considered as those that reduce fitness, sometimes due in part to the increased metabolic expenditure required to sustain life. Direct adaptation to a stressor is expected to increase fitness and reduce maintenance metabolism, with the latter leading to increased biomass production. In this study, we test the general hypothesis that such adaptation to one stressor can preadapt organisms to novel stressful environments. Six lines of Escherichia coli propagated for 2,000 generations at 41-42 degrees C (42 group), a stressful temperature, were compared to six control lines propagated for 2,000 generations at 37degrees C (37 group) and to the common ancestor of both groups. We assayed biovolume yield (a measure of growth efficiency) and competitive fitness in the 42 group's selective high temperature environment as well as five novel stressful environments-acid, alkali, ethanol, high osmolarity and peroxide. As previously reported, at high temperature the 42 group had both higher yield and fitness than the 37 group and ancestor. In the novel environments, the 42 group generally produced yields higher than the 37 group (and marginally higher than the ancestor), but we found no differences in competitive fitness among the 37 and 42 groups and the ancestor. We also found that the performance of lines within groups was not correlated across stressful environments for either yield or relative fitness. Because previous adaptation to one stressor did not improve our measure of Darwinian fitness in novel stressful environments, we conclude that the 42 group shows no useful pre-adaptation, or cross-tolerance, to these types of environments.

Adaptation, Physiological↗

PubMed2001

Genetic architecture of thermal adaptation in Escherichia coli.

Elucidating the genetic basis of adaptation on a genomewide scale has evaded biologists, but complete genome sequences and DNA high-density array technology make genomewide surveys more tractable. Six lines of Escherichia coli adapted for 2,000 generations to a stressful high temperature of 41.5 degrees C were examined on a genomewide scale for duplication/deletion events by using DNA high-density arrays. A total of five duplication and deletion events were detected. These five events occurred in three of the six lines, whereas the remaining three lines contained no detectable events. Three of the duplications were at 2.85 Mb of the E. coli chromosome, providing evidence for the replicability of the adaptation to high temperature. Four candidate genes previously shown to play roles in stress and starvation survival were identified in the region of common duplication. Expression of the two candidate genes examined is elevated over expression levels in the ancestral lines or the lines without the duplication. In the two cases where the duplication at 2.85 Mb has been further characterized, the timing of the genome reorganization is coincident with significant increases in relative fitness. In both of these cases, the model for the origin of the duplication is a complex recombination event involving insertion sequences and repeat sequences. These results provide additional evidence for the idea that gene duplication plays an integral role in adaptation, specifically as a means for gene amplification.

Adaptation, Physiological↗

PubMed2001

Postprandial exercise: prioritization or additivity of the metabolic responses?

Monitor lizards (Varanus exanthematicus) were used to examine the prioritization or additivity of the metabolic responses associated with exercise and digestion, either of which can elevate metabolic rate independently. Rates of oxygen consumption (V(O2)) and ventilation (V(E)) were measured in lizards during fasting exercise, postprandial rest and postprandial exercise. In fasting animals, V(O2) increased with walking speed to a maximal value of 15.9 ml O(2)kg(-1)min(-1) at 1.25 km h(-1). Postprandial resting metabolic rate was elevated significantly above fasting levels (4.1 versus 2.0 ml O(2)kg(-1)min(-1)). During postprandial exercise, V(O2) increased to a maximal value of 18.8 ml O(2)kg(-1)min(-1) at 1.25 km h(-1). At every level of exercise, V(O2) was significantly higher in postprandial animals by a similar increment; the maximal rate of oxygen consumption was significantly increased by 18% in postprandial individuals. Maximal V(E) did not differ in fasting and postprandial animals and, therefore, the greater V(O2)(max) of postprandial animals cannot be attributed to a higher ventilation rate. Air convection requirement (V(E)/V(O2)) is significantly lower in postprandial animals at rest and at all levels of exercise, indicating a relative hypoventilation and increased pulmonary oxygen extraction efficiency. We suggest that this increased oxygen extraction may be due to decreased cardiopulmonary shunts and/or to lower mixed venous oxygen content. The data unequivocally support an additivity model rather than prioritization models for the allocation of elevated metabolic rate: the postprandial metabolic increment is not suspended during exercise, but rather is added onto the cost of exercise. It is clear that fasting exercise did not elicit truly maximal levels of cardiopulmonary oxygen transport in these animals, indicating problems for design models that make this assumption.

Animals↗

PubMed2001

Evolutionary adaptation to temperature. VIII. Effects of temperature on growth rate in natural isolates of Escherichia coli and Salmonella enterica from different thermal environments.

Are enteric bacteria specifically adapted to the thermal environment of their hosts? In particular, do the optimal temperatures and thermal niches of the bacterial flora reflect seasonal, geographic, or phylogenetic differences in their hosts' temperatures? We examined these questions by measuring the relationship between the temperature-dependent growth rates of enteric bacteria in a free-living ectothermic host. We sampled two species of enteric bacteria (Escherichia coli and Salmonella enterica) from three natural populations of slider turtles (Trachemys scripta elegans) seasonally over two years. Despite pronounced differences in turtle body temperatures at different seasons and in different locations, we found no evidence that the thermal growth profiles of these bacteria mirrored this variation. Optimal temperatures and maximal growth rates in rich medium were nearly the same for both bacterial species (35-36 degrees C, 2.5 doublings per hour). The thermal niche (defined as the range of temperatures over which 75% of maximal growth rate occurred) was slightly higher for E. coli (28.5-41.0 degrees C) than for S. enterica (27.7-39.8 degrees C), but the niche breadth was about the same for both. We also measured the thermal dependence of growth rate in these same bacterial species isolated from mammalian hosts. Both bacterial species had temperatures of maximal growth and thermal niches that were about 2 degrees C higher than those of their respective conspecifics sampled from turtles; niche breadths were not different. These data suggest that these bacterial species are thermal generalists that do not track fine-scale changes in their thermal environments. Even major differences in body temperatures, as great as those between ectothermic and endothermic hosts, may result in the evolution of rather modest changes in thermal properties.

Adaptation, Physiological↗

PubMed2001

Evolution of thermal dependence of growth rate of Escherichia coli populations during 20,000 generations in a constant environment.

Twelve experimental populations of the bacterium Escherichia coli evolved for 20,000 generations in a defined medium at 37 degrees C. We measured their maximum growth rates across a broad range of temperatures and at several evolutionary time points to quantify the extent to which they became thermal specialists with diminished performance at other temperatures. We also sought to determine whether antagonistic pleiotropy (genetic trade-offs) or mutation accumulation (drift decay) was primarily responsible for any thermal specialization. Populations showed consistent improvement in growth rate at moderate temperatures (27-39 degrees C), but tended to have decreased growth rate at both low (20 degrees C) and high (41-42 degrees C) temperatures. Most loss occurred early in the experiment, when adaptation was most rapid. This dynamic is predicted by antagonistic pleiotropy but not by mutation accumulation. Several populations evolved high mutation rates due to defects in their DNA repair, but they did not subsequently undergo a greater decrease in growth rate at thermal extremes than populations that retained low mutation rates, contrary to the acceleration of decay predicted by mutation accumulation. Antagonistic pleiotropy therefore is more likely to be responsible for the evolution of thermal specialization observed in maximum growth rate.

Adaptation, Physiological↗

PubMed2000

An experimental test of the thermoregulatory hypothesis for the evolution of endothermy.

The thermoregulatory hypothesis proposes that endothermy in mammals and birds evolved as a thermoregulatory mechanism per se and that natural selection operated directly to increase body temperature and thermal stability through increments in resting metabolic rate. We experimentally tested this hypothesis by measuring the thermoregulatory consequences of increased metabolic rate in resting lizards (Varanus exanthematicus). A large metabolic increment was induced by feeding the animals and consequent changes in metabolic rate and body temperature were monitored. Although metabolic rate tripled at 32 degrees C and quadrupled at 35 degrees C, body temperature rose only about 0.5 degrees C. The rate of decline of body temperature in a colder environment did not decrease as metabolic rate increased. Thus, increasing the visceral metabolic rate of this ectothermic lizard established neither consequential endothermy nor homeothermy. These results are inconsistent with a thermoregulatory explanation for the evolution of endothermy.

Air↗

PubMed2000

Patterns of cardiovascular and ventilatory response to elevated metabolic states in the lizard Varanus exanthematicus.

The principal function of the cardiopulmonary system is the precise matching of O(2) and CO(2) transport to the metabolic requirements of different tissues. In some ecothermic vertebrates (amphibians and reptiles), vdot (O2) increases dramatically following feeding. Factorial increments in vdot (O2) range from 1.7 to 44 times above resting rates, and in some cases vdot (O2) approaches or even exceeds values measured during physical activity. There is virtually no information on the cardiopulmonary response during the postprandial period in these animals or how the pattern of cardiopulmonary support compares with that during activity. In our experiments, pulmonary ventilation ( vdot e), heart rate (fh), systemic blood flow ( qdot (sys)), rate of oxygen consumption ( vdot (O2)) and rate of carbon dioxide production ( vdot (CO2)) were measured at 35 degrees C in the lizard Varanus exanthematicus for 24 h prior to the ingestion of meals of various sizes and measured continuously for up to 72 h during the postprandial period. The results of this study were compared with previously published values for treadmill exercise in the same experimental animals. The change in fh and stroke volume (V(S)) for a given increment in vdot (O2) did not differ during exercise and digestion. In contrast, the ventilatory response was very dependent on the nature of the elevated metabolic state. During digestion, an increase in vdot (O2) resulted in a relative hypoventilation in comparison with resting values, whereas hyperventilation characterized the response during activity. During exercise, breathing frequency (f) increased 10- to 40-fold above resting values accompanied by large reductions in tidal volume (V(T)). In contrast, postprandial increases in vdot (O2) resulted in relatively minor changes in f and V(T) almost doubled. These results indicate that, in these lizards, the cardiac response to elevated vdot (O2) is stereotyped, the response being predictable irrespective of the source of the metabolic increment. In contrast, the ventilatory response is flexible and state-dependent, not only in pattern but also in its frequency and volume components.

Animals↗

PubMed2000

Ventilatory and cardiovascular responses of a python (Python molurus) to exercise and digestion.

To investigate the potential limiting steps of peak metabolic rates, we examined gas exchange rates ( vdot (O2), vdot (CO2)), respiratory exchange ratio (RER), breathing frequency, tidal volume, minute ventilation volume (V.e) as well as the heart rate, systemic blood flow and stroke volume of Burmese pythons (Python molurus) while fasting at rest, exercising, digesting and exercising while digesting. All measured variables increased significantly during exercise (crawling at 0.4 km h(-)(1) and at vdot (O2max)), highlighted by a 17-fold increase in vdot (CO2) and a 24-fold increase in V.e. During the digestion of a meal equivalent to 25 % of the snake's body mass, pythons responded with increases in vdot (O2) and heart rate similar to those experienced during exercise, along with a 4.5-fold increase in systemic blood flow. Interestingly, pythons hyperventilated while exercising, whereas they hypoventilated during digestion. The combined demands of exercise and digestion resulted in significantly higher vdot (O2), vdot (CO2), breathing frequency and heart rate than during either exercise or digestion alone. Evidently, the capacities of the ventilatory and cardiovascular systems to transport oxygen to locomotor muscles are not a limiting factor in the attainment of peak metabolic rates during exercise in pythons

Animals↗

PubMed2000

Effects of feeding on arterial blood gases in the American alligator Alligator mississippiensis.

Reptiles habitually ingest large meals at infrequent intervals, leading to changes in acid-base status as the net secretion of acid to the stomach causes a metabolic alkalosis (the alkaline tide). In chronically cannulated and undisturbed amphibians and reptiles, the pH changes in arterial blood are, nevertheless, reduced by a concomitant respiratory acidosis (increased P(CO2) caused by a relative hypoventilation). Alligators (Alligator mississippiensis) have been reported to exhibit exceptionally large increases in plasma [HCO3(-)] following feeding, but these studies were based on blood samples obtained by cardiac puncture, so stress and disturbance may have affected the blood gas levels. Furthermore, crocodilian haemoglobin is characterised by a unique binding of HCO3(-) that act to reduce blood oxygen-affinity, and it has been proposed that this feature safeguards oxygen offloading by counteracting pH effects on blood oxygen-affinity. Therefore, to study acid-base regulation and the interaction between the alkaline tide and oxygen transport in more detail, we describe the arterial blood gas composition of chronically cannulated and undisturbed alligators before and after voluntary feeding (meal size 7.5+/-1% of body mass). Digestion was associated with an approximately fourfold increase in metabolic rate (from 0.63+/-0.04 to 2.32+/-0.24 ml O(2) min(-1)kg(-1)) and was accompanied by a small increase in the respiratory gas exchange ratio. The arterial P(O2) of fasting alligators was 60.3+/-6.8 mmHg (1 mmHg = 0.133 kPa) and reached a maximum of 81.3+/-2.7 mmHg at 96 h following feeding; there was only a small increase in lactate levels, so the increased metabolic rate seems to be entirely aerobic. Plasma [HCO3(-)] increased from 24.4+/-1.1 to 36.9+/-1.7 mmol l(-1) (at 24 h), but since arterial P(CO2) increased from 29.0+/-1.1 to 36.8+/-1.3 mmHg, arterial pH remained virtually unaffected (changing from 7.51+/-0.01 to 7.58+/-0.01 at 24 h). The changes in plasma [HCO3(-)] were mirrored by equimolar reductions in plasma [Cl(-)]. The in vitro blood oxygen-affinity was reduced during the post-prandial period, whereas the estimated in vivo blood oxygen-affinity remained virtually constant. This supports the view that the specific HCO3(-) effect prevents an increased blood oxygen-affinity during digestion in alligators.

Alligators and Crocodiles↗

PubMed1997

Phenotypic and evolutionary adaptation of a model bacterial system to stressful thermal environments.

We studied both phenotypic and evolutionary adaptation to various thermal environments using the bacterium Escherichia coli as an experimental model system. We determined that 42 degrees C was stressful to a bacterial clone adapted to 37 degrees C, based on reductions in both absolute and competitive fitness, as well as induction of a heat stress response. This clone was also used to found replicated populations that were propagated for thousands of generations under several different thermal regimes, including 42 degrees C. Evolutionary adaptation of the populations to 42 degrees C resulted in an increase in both absolute and relative fitness at that temperature, measured respectively as an increase in the number of descendants (and their biovolume) and in competitive ability relative to the ancestral clone. The replicated experimental lineages achieved their evolutionary improvement by several distinct pathways, which produced differential preadaptation to a non-stressful nutrient environment. Adaptation to this stressful temperature entailed neither a change in the ancestral thermal niche nor any pronounced trade-offs in fitness within the thermal niche, contrary to a priori predictions. This study system was several important advantages for evaluating hypotheses concerning the effects of stress on phenotypic and evolutionary adaptation, including the ability to obtain lineages that have evolved in controlled and defined environments, to make direct measurements of fitness and to quantify the degree of stress imposed by different environments.

Adaptation, Physiological↗

PubMed1995

Experimental tests of the roles of adaptation, chance, and history in evolution.

The contributions of adaptation, chance, and history to the evolution of fitness and cell size were measured in two separate experiments using bacteria. In both experiments, populations propagated in identical environments achieved similar fitnesses, regardless of prior history or subsequent chance events. In contrast, the evolution of cell size, a trait weakly correlated with fitness, was more strongly influenced by history and chance.

Adaptation, Physiological↗

PubMed1994

Temperature acclimation and competitive fitness: an experimental test of the beneficial acclimation assumption.

Phenotypic acclimation is generally assumed to confer an advantage in the environment that stimulates the response. To test this beneficial acclimation assumption explicitly, we investigated the consequences of temperature acclimation for the fitness of Escherichia coli at two temperatures, 32 degrees C and 41.5 degrees C. Both temperatures permit growth and long-term persistence of the genotypes in serial culture. We found that prior acclimation to 32 degrees C, relative to acclimation to 41.5 degrees C, enhanced fitness at 32 degrees C, consistent with the assumption. But prior acclimation to 41.5 degrees C actually reduced fitness at 41.5 degrees C, relative to acclimation to 32 degrees C. Hence, the assumption that acclimation always confers an advantage is demonstrated to be false. Acclimation to 41.5 degrees C did, however, improve survival at 50 degrees C, a lethal temperature. This protective response has been shown to be associated with the induction of stress proteins. The reduced competitive fitness caused by acclimation at 41.5 degrees C may reflect a physiological burden associated with expression of stress proteins when they are not needed to prevent lethal damage. Whatever the cause, acclimation to the higher temperature decreased competitive fitness at that temperature.

Acclimatization↗

PubMed1994

Modeling red muscle power output during steady and unsteady swimming in largemouth bass.

We recorded electromyograms of slow-twitch (red) muscle fibers and videotaped swimming in the largemouth bass (Micropterus salmoides) during cruise, burst-and-glide, and C-start maneuvers. By use of in vivo patterns of stimulation and estimates of strain, in vitro power output was measured at 20 degrees C with the oscillatory work loop technique on slow-twitch fiber bundles from the midbody area near the soft dorsal fin. Power output increased slightly with cycle frequency to a plateau of approximately 10 W/kg at 3-5 Hz, encompassing the normal range of tail-beat frequencies for steady swimming (approximately 2-4 Hz). Power output declined at cycle frequencies simulating unsteady swimming (burst-and-glide, 10 Hz; C-start, 15 Hz). However, activating the muscle at 10 Hz did significantly increase the net work done compared with the work produced by the inactive muscle (work done by the viscous and elastic components). Thus this study provides further insight into the apparently paradoxical observation that red muscle can contribute little or no power and yet continues to show some recruitment during unsteady swimming. Comparison with published values of power requirements from oxygen consumption measurements indicates a limit to steady swimming speed imposed by the maximum power available from red muscle.

Animals↗