Anaemia adjusts the aerobic physiology of snapper (Pagrus auratus) and modulates hypoxia avoidance behaviour during oxygen choice presentations.

The effect of altered oxygen transport potential on behavioural responses to environmental hypoxia was tested experimentally in snapper, Pagrus auratus, treated with a haemolytic agent (phenylhydrazine) or a sham protocol. Standard metabolic rate was not different between anaemic and normocythaemic snapper (Hct=6.7 and 25.7 g dl(-1), respectively), whereas maximum metabolic rate, and hence aerobic scope (AS), was consistently reduced in anaemic groups at all levels of water P(O(2)) investigated (P<0.01). This reduction of AS conferred a higher critical oxygen limit (P(crit)) to anaemic fish (8.6±0.6 kPa) compared with normocythaemic fish (5.3±0.4 kPa), thus demonstrating reduced hypoxic tolerance in anaemic groups. In behavioural choice experiments, the critical avoidance P(O(2)) in anaemic fish was 6.6±2.5 kPa compared with 2.9±0.5 kPa for controls (P<0.01). Behavioural avoidance was not associated with modulation of swimming speed. Despite differences in physiological and behavioural parameters, both groups avoided low P(O(2)) just below their P(crit), indicating that avoidance was triggered consistently when AS limits were reached and anaerobic metabolism was unavoidable. This was confirmed by high levels of plasma lactate in both treatments at the point of avoidance. This is the first experimental demonstration of avoidance behaviour being modulated by internal physiological state. From an ecological perspective, fish with disturbed oxygen delivery potential arising from anaemia, pollution or stress are likely to avoid environmental hypoxia at a higher P(O(2)) than normal fish.

Characterization of the hemoglobins of the Australian lungfish Neoceratodus forsteri (Krefft).

We examined for the first time the hemoglobin components of the blood of the Australian lungfish, Neoceratodus forsteri and their functional responses to pH and the allosteric modulators adenosine triphosphate (ATP), guanosine triphosphate (GTP), 2,3-bisphosphoglyceric acid (BPG) and inositol hexaphosphate (IHP) at 25 degrees C. Lysates prepared from stripped, unfractionated hemolysate produced sigmoidal oxygen equilibrium curves with high oxygen affinity (oxygen partial pressure required for 50% hemoglobin saturation, p(50)=5.3 mmHg) and a Hill coefficient of 1.9 at pH 7.5. p(50) was 8.3 and 4.5 mmHg at pH 6 and 8, respectively, which corresponded to a modest Bohr coefficient (Delta log p(50)/Delta pH) of -0.13. GTP increased the pH sensitivity of oxygen binding more than ATP, such that the Bohr coefficient was -0.77 in the presence of 2 mmol L(-1) GTP. GTP was the most potent regulator of hemoglobin affinity, with concentrations of 5 mmol L(-1) causing an increase in p(50) from 5 to 19 mm Hg at pH 7.5, while the order of potency of the other phosphates was IHP>ATP>BPG. Three hemoglobin isoforms were present and each contained both alpha and beta chains with distinct molecular weights. Oxygen affinity and pH-dependence of isoforms I and II were essentially identical, while isoform III had a lower affinity and increased pH-dependence. The functional properties of the hemoglobin system of Neoceratodus appeared consistent with an active aquatic breather adapted for periodic hypoxic episodes.

Characterization of the hemoglobins of the neonatal brushtailed possum Trichosurus vulpecula (Kerr): evidence for a highly cooperative, aggregated isoform of hemoglobin.

The red blood cells of the neonatal brushtailed possum exhibit unusually strong cooperativity at high levels of oxygen saturation (n=5.4) which appear to arise from a concentration dependent aggregation of one of the neonatal hemoglobin isoforms. Red blood cells from neonatal pouched young exhibit a Bohr factor of -0.36. Stripped hemolysate is sensitive to added 2,3-bisphosphoglycerate (BPG) (apparent binding constant K=35 micromol L(-1)) and ATP (K=180 micromol L(-1)), but is largely insensitive towards chloride ions. Five isoforms of non-adult hemoglobin were identified using isoelectric focusing. Mass spectrometry indicated that two early isoforms contain alpha chains identical to the adult alpha chain. The remaining three isoforms are composed of identical alpha type and beta type gene products, but differ in their isoelectric points due to differential post-translational modification.

Ontogenetic scaling of fish metabolism in the mouse-to-elephant mass magnitude range.

Intraspecific or ontogenetic analyses of mass-metabolism relationships do not often conform to the same allometric correlations as those seen in interspecific analyses. A commonly cited reason for this discrepancy is that ontogenetic studies examine smaller mass ranges than interspecific studies, and are therefore not statistically comparable. In this study the metabolic rate of yellowtail kingfish was measured from 0.6 mg-2.2 kg, a mass range comparable to that between a mouse and an elephant. Linear regression of the log transformed data resulted in a scaling exponent of 0.90 and high correlation coefficient. Statistical and information theory comparisons of three other models showed that a segmented linear regression and curvilinear quadratic function were an improvement over a simple linear regression. This confirmed previous observations that the metabolic scaling exponent of fish changes during ontogeny. Ammonia excretion rates were also measured and scaled linearly with an exponent of 0.87. The data showed that the metabolism of yellowtail kingfish during ontogeny did not scale with the commonly cited 2/3 or 3/4 mass exponent. This demonstrates that differences between interspecific and ontogenetic allometries are not necessarily statistical artefacts.

Characterization of the hemoglobins of the adult brushtailed possum, Trichosurus vulpecula (Kerr) reveals non-genetic heterogeneity.

The hemoglobins contained within the red blood cells of the adult brushtail possum exhibited cooperative (n=2.6) oxygen binding curves with an associated p50 of 38 mm Hg at pH 7.4 and a large Bohr effect (-0.60). Stripped hemolysate showed a Bohr effect of -0.27, and was sensitive to added DPG (K=56 micromol L(-1)), ATP (K=130 micromol L(-1)), and chloride ions. Four isoforms of hemoglobin were identified using isoelectric focusing. Mass spectrometry indicated that all four isoforms most likely represent the same gene products which have differentially undergone post-translational deamidation and glutathionylation. The oxygen binding characteristics of three isolated isohemoglobins have been determined.

Continuous measurement of oxygen tensions in the air-breathing organ of Pacific tarpon (Megalops cyprinoides) in relation to aquatic hypoxia and exercise.

The Pacific tarpon is an elopomorph teleost fish with an air-breathing organ (ABO) derived from a physostomous gas bladder. Oxygen partial pressure (PO(2)) in the ABO was measured on juveniles (238 g) with fiber-optic sensors during exposure to selected aquatic PO(2) and swimming speeds. At slow speed (0.65 BL s(-1)), progressive aquatic hypoxia triggered the first breath at a mean PO(2) of 8.3 kPa. Below this, opercular movements declined sharply and visibly ceased in most fish below 6 kPa. At aquatic PO(2) of 6.1 kPa and swimming slowly, mean air-breathing frequency was 0.73 min(-1), ABO PO(2) was 10.9 kPa, breath volume was 23.8 ml kg(-1), rate of oxygen uptake from the ABO was 1.19 ml kg(-1) min(-1), and oxygen uptake per breath was 2.32 ml kg(-1). At the fastest experimental speed (2.4 BL s(-1)) at 6.1 kPa, ABO oxygen uptake increased to about 1.90 ml kg(-1) min(-1), through a variable combination of breathing frequency and oxygen uptake per breath. In normoxic water, tarpon rarely breathed air and apparently closed down ABO perfusion, indicated by a drop in ABO oxygen uptake rate to about 1% of that in hypoxic water. This occurred at a wide range of ABO PO(2) (1.7-26.4 kPa), suggesting that oxygen level in the ABO was not regulated by intrinsic receptors.

Whole blood-oxygen binding properties of four cold-temperate marine fishes: blood affinity is independent of pH-dependent binding, routine swimming performance, and environmental hypoxia.

The relationship between whole blood-oxygen affinity (P(50)) and pH-dependent binding (i.e., cooperativity and the Bohr [ Phi ] and Root effects) was examined statistically under standardized conditions (10.0 degrees Celsius) in four unrelated cold-temperate marine fishes that differ widely in their swimming performance and their expected responses to hypoxia: cod (Gadus morhua), herring (Clupea harengus), mackerel (Scomber scombrus), and plaice (Pleuronectes platessa). An unexpected difference in blood-oxygen affinity was found (herring>plaice>mackerel>cod), and this was independent of both swimming performance and the predicted low O(2) response of each species. The ecotype of the four marine species was also unrelated to pH-dependent binding because no difference in the Bohr effect was apparent ( Phi varied insignificantly from -0.90 to -1.06), and differences in the magnitude of the cooperative binding reaction were associated only with the presence of the Root effect. Although several reviews propose a generalized link between blood-oxygen affinity and pH-dependent binding, our results advise against overestimating the adaptive functional properties of hemoglobin across unrelated species.