SUMMARY European eels were exposed for 6 weeks to water CO2 partial pressures (PCO2) from ambient (approx. 0.8 mmHg), through 15±1 mmHg and 30±1 mmHg to 45±1 mmHg in water with a total hardness of 240 mg l–1 as CaCO3, pH 8.2, at 23±1°C. Arterial plasma PCO2 equilibrated at approximately 2 mmHg above water PCO2 in all groups, and plasma bicarbonate accumulated up to 72 mmol l–1 in the group at a water PCO2 of 45 mmHg. This was associated with an equimolar loss of plasma Cl–, which declined to 71 mmol l–1 at the highest water PCO2. Despite this, extracellular acid–base compensation was incomplete; all hypercapnic groups tolerated chronic extracellular acidoses and reductions in arterial blood O2 content (CaO2), of progressive severity with increasing PCO2. All hypercapnic eels, however, regulated the intracellular pH of heart and white muscle to the same levels as normocapnic animals. Hypercapnia had no effect on such indicators of stress as plasma catecholamine or cortisol levels, plasma osmolality or standard metabolic rate. Furthermore, although CaO2 was reduced by approximately 50% at the highest PCO2, there was no effect of hypercapnia on the eels' tolerance of hypoxia, aerobic metabolic scope or sustained swimming performance. The results indicate that, at the levels tested, chronic hypercapnia was not a physiological stress for the eel, which can tolerate extracellular acidosis and extremely low Cl– levels while compensating tissue intracellular pH, and which can meet the O2 requirements of routine and active metabolism despite profound hypoxaemia.

Fish represent the oldest and most diverse classes of vertebrates, comprising around the 48% of the known member species in the subphylum Vertebrata. There are many scientific fields that use fish as models in research, including respiratory and cardiovascular research, cell culture, ecotoxicology, ageing, pharmacological and genetic studies.

European eels (Anguilla anguilla, L.) were fed on a commercial diet supplemented either with 15% by dry feed weight of menhaden oil (MO), an oil rich in highly unsaturated fatty acids of the n-3 series (n-3 HUFA), or with 15% by dry feed weight of coconut oil (CO), an oil composed primarily of saturated fatty acids (SFA). Following 90 days of feeding, the mean final masses of eels fed the two different oil supplements were similar, and higher than the mean final mass of a group fed the commercial diet alone. The diets created two distinct phenotypes of eels, distinguished by the fatty acid (FA) composition of their tissue lipids. Eels fed MO had significantly more total n-3 FA and n-3 HUFA in muscle and liver lipids than did eels fed CO, leading to higher n-3/n-6 and eicosapentaenoic acid/arachidonic acid ratios in the MO group. Measurements of O2 uptake (MO2) revealed that the MO group had a significantly lower routine metabolic rate (RMR) than the CO group. When exposed to progressive hypoxia, both groups regulated MO2 at routine normoxic levels until critical water O2 partial pressures that were statistically similar (9.62±1.08 kPa in MO versus 7.57±1.07 kPa in CO), beyond which they showed a reduction in MO2 below RMR. The MO group exhibited a significantly lower MO2 than the CO group throughout hypoxic exposure, but the percentage reductions in MO2 below their relative RMR were equal in both groups. During recovery to normoxia, both groups exhibited an increase in MO2 to rates significantly higher than their RMR. Throughout recovery, MO2 was significantly lower in the MO group compared with the CO group, but the percentage increases in MO2 relative to RMR were equal in both. During progressive hypoxia, neither group exhibited a marked ventilatory reflex response, both showed similar reductions in blood O2 partial pressure and content, and similar increases in plasma lactate. The results indicate that, although the n-3 HUFA-enriched MO group had a significantly lower routine metabolic rate than the CO group, the difference in aerobic metabolism did not influence the European eel's homeostatic regulation of MO2 in hypoxia.

Nile tilapia (Oreochromis niloticus) were infused with ammonium salts, acid, and base to investigate the effects of changes in arterial plasma total ammonia content (Tamm) and pH (pHa) on plasma urea-nitrogen (urea-N) levels and urea-N excretory fluxes (Jurea-N). The tilapia did not possess a functional hepatic ornithine urea-cycle (no significant carbamyl phosphate synthetase III activity). Infused substances were dissolved in a saline vehicle and injected twice (5 mL kg-1), the first infusion to "prime" the animal and promote a more marked response to the second infusion, given 2.5 h later. The results reported are those of the second infusion. Infusion of 200 mM NH4Cl increased Tamm, reduced pHa, and increased plasma urea-N and Jurea-N. Two hundred mM NH4HCO3 increased Tamm and arterial plasma total CO2 content (TaCO2), reduced pHa, and increased Jurea-N. Fifty mM HCl reduced pHa but had no effects on urea dynamics. Fifty mM NaOH increased pHa, plasma urea-N levels, and Jurea-N. Two hundred mM NaHCO3 increased pHa, TaCO2, plasma urea-N levels, and Jurea-N. Infusion of the saline vehicle was without effect. The results indicate that ammonia loading and plasma alkalosis both stimulate urea excretion in uricolytic fish. The responses to hyperammonemia or alkalosis were not modified when combined with elevated plasma bicarbonate levels.

Summary This paper reviews evidence that the fatty acid composition of dietary lipids influences the respiratory and cardiovascular physiology of Adriatic sturgeon (Acipenser naccarii) and, thereby, their tolerance of the stress of hypoxia. Sturgeon fed a commercial diet enriched in fish oil (menhaden oil as 15% of dry feed weight), with an elevated content of highly unsaturated fatty acids of the ω3 series (ω3 HUFA), had a significantly lower standard metabolic rate (SMR) and routine oxygen consumption (Mo2) than those fed a diet enriched with the same quantity of hydrogenated coconut oil, with an elevated content of saturated fatty acids (SFA). Both groups grew equally well. As a result of this difference in aerobic metabolism, sturgeon fed the ω3 HUFA and SFA responded differently when exposed to hypoxic challenges, those fed ω3 HUFA appearing more hypoxia-tolerant. Sturgeon fed ω3 HUFA exhibited no significant reflex hyperventilation when exposed to mild, moderate or deep hypoxia (30 min at water O2 partial pressures of 10.8, 6.6 and 4.6 kPa, respectively), no hypoxic depression of spontaneous activity during 3h in mild hypoxia, and no depression of Mo2 during 3h in moderate hypoxia, unlike sturgeon fed SFA. The diets also influenced the performance of isolated hearts in vitro. Hearts from fish fed ω3 HUFA maintained maximum in vitro cardiac power output unchanged when oxygen supply was reduced (O2 content from 2.3 to 0.7 vol.%), unlike hearts from sturgeon fed SFA. Overall, the results indicate that dietary fatty acid composition can influence tolerance of hypoxia in sturgeon, through effects on SMR. When compared to sturgeon fed SFA, those fed ω3 HUFA had lower SMR and were more tolerant of hypoxia, with effects both on the whole animal and on the isolated heart.

The effects of exhaustive exercise on O2 consumption (ṀO2), waste nitrogen (ammonia and urea) excretion, and on lactate, ammonia and water content of white muscle and liver, were determined in tilapia fed a diet enriched either in long chain polyunsaturated fatty acids of the ω3 series (ω3 LCPUFA) as menhaden oil (menhaden oil diet; MOD) or in saturated fatty acids (SFA) as coconut oil (coconut oil diet; COD), and acclimated to three different temperatures (16°, 23° and 33°C). At all temperatures, exhaustive exercise elicited an increase in postexercise ṀO2 and ammonia excretion rates, and in white muscle lactate and ammonia levels. There were no differences between the two dietary groups in the total amount of O2 consumed and muscle lactate accumulated, but at 23°C and 33°C exhaustive exercise stimulated a significantly greater increase in ammonia excretion in COD as compared with MOD tilapia. The magnitude of the difference in postexercise ammonia excretion between COD and MOD animals increased with increases in environmental temperature, being greater at 33°C than at 23°C. The increased ammonia excretion observed in the COD group was not a result of differences in white muscle ammonia or water content following exhaustive exercise. The data indicate that diets enriched in ω3 LCPUFA are beneficial in that tilapia fed these diets exhibited reduced ammonia excretion following exhaustive exercise, compared with tilapia fed diets enriched in SFA.

Dietary polyunsaturated fatty acids (PUFA) of the n-3 series that have beneficial effects on mammalian heart function are typically found at high levels in fish tissues. The effects of dietary fatty acid composition on cardiac function were investigated in the sturgeon. When compared with sturgeon maintained for 1 yr on a diet enriched with saturated fatty acids (SFA) (the coconut oil-supplemented diet, COD), sturgeon maintained on a diet enriched with n-3 PUFA (the fish oil-supplemented diet, FOD) had higher myocardial 20:5(n-3) and lower 20:4(n-6) content with a consequent decrease in the n-6-to-n-3 ratio (from 0.86 to 0.25) and a lower intrinsic in vitro heart rate (22.0 +/- 1.5 vs. 29.9 +/- 1.0 beats/min) and cardiac power output (PO) (0.33 +/- 0.08 vs. 0.48 +/- 0.03 mW/g), but had a greater in vitro scope for cardiac work (almost twice the maximal-to-basal PO ratio). Reducing the oxygen supply to the hearts significantly decreased, by approximately 40%, the maximal in vitro PO in the COD group of animals but had no effect in the FOD group. These differences in performance were not reflected in heart rate or blood pressure in vivo, either in normoxia or hypoxia. Addition of vitamin E as an antioxidant to the diets reduced intrinsic heart rate by approximately 25% but did not influence the effects (dietary fatty acid composition on in vitro cardiac performance. The results indicate that dietary n-3 PUFA can have beneficial effects on the resistance of the fish heart to environmental stressors such as hypoxia.

Cardioventilatory responses to hypoxia, the O2 chemoreceptor stimulant sodium cyanide (NaCN), and intra-arterial injection of atropine, noradrenaline and DL-propranolol were investigated in the adriatic sturgeon. Hypoxia elicited a bradycardia and hyperventilation. 1 mg NaCN added to water entering the buccal cavity stimulated a transient bradycardia but intra-arterial infusion of 150 μg NaCN did not, indicating that hypoxic bradycardia is controlled by chemoreceptors sensitive only to water O2 levels. NaCN stimulated hyperventilation both when added to the water and when infused intra-arterially, indicating that hypoxic hyperventilation is controlled by chemoreceptors sensitive to both internal and external milieux. Atropine abolished the hypoxic bradycardia and returned heart rate to normoxic values indicating that this species has no inhibitory vagal tone in normoxia. Noradrenaline stimulated ventilation, an effect abolished by DL-propranolol. Propranolol blocked ventilatory responses to intra-arterial infusion of NaCN whereas responses to NaCN added to the water remained unaffected, indicating that propranolol may inhibit internally-oriented O2-chemoreceptor activity or that ventilatory responses to intra-arterial NaCN are stimulated by a release of circulating catecholamines. Cardioventilatory control systems in sturgeon are similar to those of other actinopterygians but also show some characteristics of the system described for elasmobranchs.

Adriatic sturgeon (Acipenser naccarii) were maintained on a commercial diet enriched either in long chain polyunsaturated fatty acids of the ω3 series (ω3 LCPUFA) or in saturated fatty acids (SFA). The effects of dietary fatty acid composition on spontaneous locomotor activity in normoxia and hypoxia (O2 tension = 10.5 ± 0.8 kPa), and on oxygen consumption (MO2) in normoxia, in hypoxia (O2 tension = 6.6 ± 0.8 kPa) and during recovery were then investigated. The effects of adding supplementary vitamin E to the fat-enriched diets were also studied.