Ben Speers-Roesch

The unusual energy metabolism of elasmobranch fishes

The unusual energy metabolism of elasmobranchs is characterized by limited or absent fatty acid oxidation in cardiac and skeletal muscle and a great reliance on ketone bodies and amino acids as oxidative fuels in these tissues. Other extrahepatic tissues in elasmobranchs rely on ketone bodies and amino acids for aerobic energy production but, unlike muscle, also appear to possess a significant capacity to oxidize fatty acids. This organization of energy metabolism is reflected by relatively low plasma levels of non-esterified fatty acids (NEFA) and by plasma levels of the ketone body ss-hydroxybutyrate that are as high as those seen in fasted mammals. The preference for ketone body oxidation rather than fatty acid oxidation in muscle of elasmobranchs under routine conditions is opposite to the situation in teleosts and mammals. Carbohydrates appear to be utilized as a fuel source in elasmobranchs, similar to other vertebrates. Amino acid- and lipid-fueled ketogenesis in the liver, the lipid storage site in elasmobranchs, sustains the demand for ketone bodies as oxidative fuels. The liver also appears to export NEFA and serves a buoyancy role. The regulation of energy metabolism in elasmobranchs and the effects of environmental factors remain poorly understood. The metabolic organization of elasmobranchs was likely present in the common ancestor of the Chondrichthyes ca. 400million years ago and, speculatively, it may reflect the ancestral metabolism of jawed vertebrates. We assess hypotheses for the evolution of the unusual energy metabolism of elasmobranchs and propose that the need to synthesize urea has influenced the utilization of ketone bodies and amino acids as oxidative fuels.

Effects of environmental hypoxia on cardiac energy metabolism and performance in tilapia

The ability of an animal to depress ATP turnover while maintaining metabolic energy balance is important for survival during hypoxia. In the present study, we investigated the responses of cardiac energy metabolism and performance in the hypoxia-tolerant tilapia (Oreochromis hybrid sp.) during exposure to environmental hypoxia. Exposure to graded hypoxia (> or =92% to 2.5% air saturation over 3.6 +/- 0.2 h) followed by exposure to 5% air saturation for 8 h caused a depression of whole animal oxygen consumption rate that was accompanied by parallel decreases in heart rate, cardiac output, and cardiac power output (CPO, analogous to ATP demand of the heart). These cardiac parameters remained depressed by 50-60% compared with normoxic values throughout the 8-h exposure. During a 24-h exposure to 5% air saturation, cardiac ATP concentration was unchanged compared with normoxia and anaerobic glycolysis contributed to ATP supply as evidenced by considerable accumulation of lactate in the heart and plasma. Reductions in the provision of aerobic substrates were apparent from a large and rapid (in <1 h) decrease in plasma nonesterified fatty acids concentration and a modest decrease in activity of pyruvate dehydrogenase. Depression of cardiac ATP demand via bradycardia and an associated decrease in CPO appears to be an integral component of hypoxia-induced metabolic rate depression in tilapia and likely contributes to hypoxic survival.

Hypoxia Tolerance in Sculpins Is Associated with High Anaerobic Enzyme Activity in Brain but Not in Liver or Muscle

We assessed hypoxia tolerance in 11 species of fish from the superfamily Cottoidea (commonly called sculpins) that are known to differ in their critical O2 tensions (Pcrit) and examined whether hypoxia tolerance correlated with larger substrate stores and higher maximal activity of enzymes associated with anaerobic adenosine triphosphate production (especially glycolysis). Among the sculpins studied, there was large variation in time to loss of equilibrium (LOE50) at torr, with values ranging between 25 and 538 min, and the variation in LOE50 was correlated with Pcrit. Our measures of time to LOE50 and Pcrit were regressed against maximal enzyme activities of lactate dehydrogenase (LDH), pyruvate kinase (PK), creatine phosphokinase (CPK), and citrate synthase (CS) as well as the concentrations of glycogen, glucose, and creatine phosphate in the brain, liver, and white muscle. In the brain, there was a phylogenetically independent relationship between Pcrit and tissue LDH, PK, CPK, and CS activities expressed relative to tissue mass. Hypoxia-tolerant sculpins (those with low Pcrit values) had higher levels of brain LDH, PK, CPK, and CS than did hypoxia-sensitive sculpins. Similarly, LOE50 regressed against brain LDH, PK, and CPK activities expressed relative to tissue mass, with the more hypoxia-tolerant species (i.e., those with higher LOE50) having higher enzyme activities. However, when the phylogenetic relationship among our sculpins was taken into account, only the relationship between hypoxia tolerance and LDH activity remained significant. When enzyme activities were expressed relative to total soluble protein in the tissue, the only relationships that remained were between brain LDH activity and Pcrit and LOE50. In liver and white muscle, there were no relationships between the measures of hypoxia tolerance and enzyme activity or metabolite content. Overall, our analysis suggests that hypoxia-tolerant sculpins maintain higher maximal activities of some of the enzymes involved in anaerobic metabolism in the brain, and this may be an adaptation to hypoxia.

Exceptional cardiac anoxia tolerance in tilapia (Oreochromis hybrid).

SUMMARY Anoxic survival requires the matching of cardiac ATP supply (i.e. maximum glycolytic potential, MGP) and demand (i.e. cardiac power output, PO). We examined the idea that the previously observed in vivo downregulation of cardiac function during exposure to severe hypoxia in tilapia (Oreochromis hybrid) represents a physiological strategy to reduce routine PO to within the heart’s MGP. The MGP of the ectothermic vertebrate heart has previously been suggested to be ∼70 nmol ATP s–1 g–1, sustaining a PO of ∼0.7 mW g–1 at 15°C. We developed an in situ perfused heart preparation for tilapia (Oreochromis hybrid) and characterized the routine and maximum cardiac performance under both normoxic (>20 kPa O2) and severely hypoxic perfusion conditions (<0.20 kPa O2) at pH 7.75 and 22°C. The additive effects of acidosis (pH 7.25) and chemical anoxia (1 mmol l–1 NaCN) on cardiac performance in severe hypoxia were also examined. Under normoxic conditions, cardiac performance and myocardial oxygen consumption rate were comparable to those of other teleosts. The tilapia heart maintained a routine normoxic cardiac output (Q) and PO under all hypoxic conditions, a result that contrasts with the hypoxic cardiac downregulation previously observed in vivo under less severe conditions. Thus, we conclude that the in vivo downregulation of routine cardiac performance in hypoxia is not needed in tilapia to balance cardiac energy supply and demand. Indeed, the MGP of the tilapia heart proved to be quite exceptional. Measurements of myocardial lactate efflux during severe hypoxia were used to calculate the MGP of the tilapia heart. The MGP was estimated to be 172 nmol ATP s–1 g–1 at 22°C, and allowed the heart to generate a POmax of at least ∼3.1 mW g–1, which is only 30% lower than the POmax observed with normoxia. Even with this MGP, the additional challenge of acidosis during severe hypoxia decreased maximum ATP turnover rate and POmax by 30% compared with severe hypoxia alone, suggesting that there are probably direct effects of acidosis on cardiac contractility. We conclude that the high maximum glycolytic ATP turnover rate and levels of PO, which exceed those measured in other ectothermic vertebrate hearts, probably convey a previously unreported anoxia tolerance of the tilapia heart, but a tolerance that may be tempered in vivo by the accumulation of acidotic waste during anoxia.

Effects of Temperature and Hydrostatic Pressure on Routine Oxygen Uptake of the Bloater (Coregonus hoyi)

We present the first measurements of routine oxygen uptake (VO2) of the bloater (Coregonus hoyi), including the effects of temperature and hydrostatic pressure. For temperature experiments, ten 24 hour trials were conducted each at 10.5°C and 7°C, using flow-through respirometry. Average routine VO2 was approximately 130 mg O2/(kg*hr) at 10.5°C and approximately 120 mg O2/(kg*hr) at 7°C. These values did not differ significantly, probably because we used temperatures spanning the thermal range of bloater, within which this species may conserve routine metabolism. Derivations of daily food ration for bloater were calculated and compared with a previous bioenergetics model. For pressure experiments, the diel vertical migrations bloater undertake in the wild were simulated using a flow-through respirometer that could be pressurized. Ten 3-day trials were conducted, consisting of an overnight acclimation to the respirometer, a 12-hour pressure regime, and a day of recovery. The pressure regime involved a compression from 1 atm (atmospheric pressure) to 4 atm over 6 hours and a subsequent decompression of the same magnitude and duration. Increases in hydrostatic pressure elicited a rise in bloater VO2 and motor activity; conversely, the subsequent decrease in hydrostatic pressure caused a return of oxygen uptake and motor activity to baseline values at 1 atm. We hypothesize that pressure-induced compression of the gas bladder explain the changes in VO2, because increased swimming (causing increased VO2) is needed to maintain station when the swimbladder is compressed.

Plasma non-esterified fatty acids of elasmobranchs : Comparisons of temperate and tropical species and effects of environmental salinity

We investigated the influence of environments with different average temperatures and different salinities on plasma NEFA in elasmobranchs by comparing species from tropical vs. cold temperate marine waters, and tropical freshwater vs. tropical marine waters. The influence of the environment on plasma NEFA is significant, especially with regard to essential fatty acids (EFA) and the n-3/n-6 ratio. n-3/n-6 ratios in tropical marine elasmobranchs were lower by two-fold or more compared with temperate marine elasmobranchs, because of higher levels of arachidonic acid (AA, 20:4n-6) and docosatetraenoic acid (22:4n-6), and less docosahexaenoic acid (DHA, 22:6n-3), in the tropical species. These results are similar to those in earlier studies on lipids in teleosts. n-3/n-6 ratios and levels of EFA were similar between tropical freshwater and tropical marine elasmobranchs. This suggests that the observation in temperate waters that marine fishes have higher levels of n-3 fatty acids and n-3/n-6 ratios than freshwater fishes may not hold true in tropical waters, at least in elasmobranchs. It also suggests that plasma NEFA are little affected by freshwater vs. seawater adaptation in elasmobranchs. Likewise, we found that plasma NEFA composition and levels were not markedly affected by salinity acclimation (2 weeks) in the euryhaline stingray Himantura signifer. However, in contrast to our comparisons of freshwater-adapted vs. marine species, the level of n-3 fatty acids and the n-3/n-6 ratio were observed to significantly decrease, indicating a potential role of n-3 fatty acids in salinity acclimation in H. signifer.

Does the physiology of chondrichthyan fishes constrain their distribution in the deep sea

ABSTRACT The deep sea is the largest ecosystem on Earth but organisms living there must contend with high pressure, low temperature, darkness and scarce food. Chondrichthyan fishes (sharks and their relatives) are important consumers in most marine ecosystems but are uncommon deeper than 3000 m and exceedingly rare, or quite possibly absent, from the vast abyss (depths >4000 m). By contrast, teleost (bony) fishes are commonly found to depths of ∼8400 m. Why chondrichthyans are scarce at abyssal depths is a major biogeographical puzzle. Here, after outlining the depth-related physiological trends among chondrichthyans, we discuss several existing and new hypotheses that implicate unique physiological and biochemical characteristics of chondrichthyans as potential constraints on their depth distribution. We highlight three major, and not mutually exclusive, working hypotheses: (1) the urea-based osmoregulatory strategy of chondrichthyans might conflict with the interactive effects of low temperature and high pressure on protein and membrane function at great depth; (2) the reliance on lipid accumulation for buoyancy in chondrichthyans has a unique energetic cost, which might increasingly limit growth and reproductive output as food availability decreases with depth; (3) their osmoregulatory strategy may make chondrichthyans unusually nitrogen limited, a potential liability in the food-poor abyss. These hypotheses acting in concert could help to explain the scarcity of chondrichthyans at great depths: the mechanisms of the first hypothesis may place an absolute, pressure-related depth limit on physiological function, while the mechanisms of the second and third hypotheses may limit depth distribution by constraining performance in the oligotrophic abyss, in ways that preclude the establishment of viable populations or lead to competitive exclusion by teleosts. Summary: Chondrichthyan fishes (sharks, rays and chimaeras) are exceedingly rare or possibly absent at abyssal depths (>4000 m), unlike bony fishes. This Commentary discusses hypotheses implicating the unusual physiology of chondrichthyans as an explanation for their scarcity at great depths.

Oxygen- and capacity-limited thermal tolerance: blurring ecology and physiology.

The Commentary by Portner, Bock and Mark ([Portner et al., 2017][1]) elaborates on the oxygen- and capacity-limited thermal tolerance (OCLTT) hypothesis. Journal of Experimental Biology Commentaries allow for personal and controversial views, yet the journal also mandates that ‘opinion and fact

Metabolic rate and rates of protein turnover in food-deprived cuttlefish, Sepia officinalis (Linnaeus 1758)

To determine the metabolic response to food deprivation, cuttlefish (Sepia officinalis) juveniles were either fed, fasted (3 to 5 days food deprivation), or starved (12 days food deprivation). Fasting resulted in a decrease in triglyceride levels in the digestive gland, and after 12 days, these lipid reserves were essentially depleted. Oxygen consumption was decreased to 53% and NH4 excretion to 36% of the fed group following 3-5 days of food deprivation. Oxygen consumption remained low in the starved group, but NH4 excretion returned to the level recorded for fed animals during starvation. The fractional rate of protein synthesis of fasting animals decreased to 25% in both mantle and gill compared with fed animals and remained low in the mantle with the onset of starvation. In gill, however, protein synthesis rate increased to a level that was 45% of the fed group during starvation. In mantle, starvation led to an increase in cathepsin A-, B-, H-, and L-like enzyme activity and a 2.3-fold increase in polyubiquitin mRNA that suggested an increase in ubiquitin-proteasome activity. In gill, there was a transient increase in the polyubiquitin transcript levels in the transition from fed through fasted to the starved state and cathepsin A-, B-, H-, and L-like activity was lower in starved compared with fed animals. The response in gill appears more complex, as they better maintain rates of protein synthesis and show no evidence of enhanced protein breakdown through recognized catabolic processes.

Maximum thermal limits of coral reef damselfishes are size dependent and resilient to near-future ocean acidification

ABSTRACT Theoretical models predict that ocean acidification, caused by increased dissolved CO2, will reduce the maximum thermal limits of fishes, thereby increasing their vulnerability to rising ocean temperatures and transient heatwaves. Here, we tested this prediction in three species of damselfishes on the Great Barrier Reef, Australia. Maximum thermal limits were quantified using critical thermal maxima (CTmax) tests following acclimation to either present-day or end-of-century levels of CO2 for coral reef environments (∼500 or ∼1000 µatm, respectively). While species differed significantly in their thermal limits, whereby Dischistodus perspicillatus exhibited greater CTmax (37.88±0.03°C; N=47) than Dascyllus aruanus (37.68±0.02°C; N=85) and Acanthochromis polyacanthus (36.58±0.02°C; N=63), end-of-century CO2 had no effect (D. aruanus) or a slightly positive effect (increase in CTmax of 0.16°C in D. perspicillatus and 0.21°C in A. polyacanthus) on CTmax. Contrary to expectations, early-stage juveniles were equally as resilient to CO2 as larger conspecifics, and CTmax was higher at smaller body sizes in two species. These findings suggest that ocean acidification will not impair the maximum thermal limits of reef fishes, and they highlight the critical role of experimental biology in testing predictions of theoretical models forecasting the consequences of environmental change. Summary: Despite a widespread perception that end-of-century ocean acidification will reduce the thermal limits of fishes, we show that critical thermal maxima of coral reef damselfishes are robust to this stressor.