Ole Brix

Climate induced temperature effects on growth performance, fecundity and recruitment in marine fish: developing a hypothesis for cause and effect relationships in Atlantic cod (Gadus morhua) and common eelpout (Zoarces viviparus)

Effects of global warming on animal distribution and performance become visible in many marine ecosystems. The present study was designed to develop a concept for a cause and effect understanding with respect to temperature changes and to explain ecological findings based on physiological processes. The concept is based on a wide comparison of invertebrate and fish species with a special focus on recent data obtained in two model species of fish. These fish species are both characterized by northern and southern distribution limits in the North Atlantic: eelpout (Zoarces viviparus), as a typical non-migrating inhabitant of the coastal zone and the cod (Gadus morhua), as a typical inhabitant of the continental shelf with a high importance for fisheries. Mathematical modelling demonstrates a clear significant correlation between climate induced temperature fluctuations and the recruitment of cod stocks. Growth performance in cod is optimal at temperatures close to 10°C, regardless of the population investigated in a latitudinal cline. However, temperature specific growth rates decrease at higher latitudes. Also, fecundity is less in White Sea than in North and Baltic Sea cod or eelpout populations. These findings suggest that a cold-induced shift in energy budget occurs which is unfavorable for growth performance and fecundity. Thermal tolerance limits shift depending on latitude and are characterized by oxygen limitation at both low or high temperatures. Oxygen supply to tissues is optimized at low temperature by a shift in hemoglobin isoforms and oxygen binding properties to lower affinities and higher unloading potential. Protective stimulation of heat shock protein synthesis was not observed. According to a recent model of thermal tolerance the downward shift of tolerance limits during cold adaptation is associated with rising mitochondrial densities and, thus, aerobic capacity and performance in the cold, especially in eurythermal species. At the same time the costs of mitochondrial maintenance reflected by mitochondrial proton leakage should rise leaving a lower energy fraction for growth and reproduction. The preliminary conclusion can be drawn that warming will cause a northern shift of distribution limits for both species with a rise in growth performance and fecundity larger than expected from the Q10 effect in the north and lower growth or even extinction of the species in the south. Such a shift may heavily affect fishing activities in the North Sea.

Haemoglobin polymorphisms affect the oxygen- binding properties in Atlantic cod populations

A major challenge in evolutionary biology is to identify the genes underlying adaptation. The oxygen-transporting haemoglobins directly link external conditions with metabolic needs and therefore represent a unique system for studying environmental effects on molecular evolution. We have discovered two haemoglobin polymorphisms in Atlantic cod populations inhabiting varying temperature and oxygen regimes in the North Atlantic. Three-dimensional modelling of the tetrameric haemoglobin structure demonstrated that the two amino acid replacements Met55β1Val and Lys62β1Ala are located at crucial positions of the α1β1 subunit interface and haem pocket, respectively. The replacements are proposed to affect the oxygen-binding properties by modifying the haemoglobin quaternary structure and electrostatic feature. Intriguingly, the same molecular mechanism for facilitating oxygen binding is found in avian species adapted to high altitudes, illustrating convergent evolution in water- and air-breathing vertebrates to reduction in environmental oxygen availability. Cod populations inhabiting the cold Arctic waters and the low-oxygen Baltic Sea seem well adapted to these conditions by possessing the high oxygen affinity Val55–Ala62 haplotype, while the temperature-insensitive Met55–Lys62 haplotype predominates in the southern populations. The distinct distributions of the functionally different haemoglobin variants indicate that the present biogeography of this ecologically and economically important species might be seriously affected by global warming.

Genetic variation and functional properties of Atlantic cod hemoglobins: introducing a modified tonometric method for studying fragile hemoglobins.

Hemoglobin polymorphism in Atlantic cod has been investigated with respect to physiological performance at 10, 15 and 20 degrees C applying a modified tonometric method for O2 equilibrium analysis with full control of the equilibrating gas mixture. The results did not indicate any dissociation of the hemoglobins by a reduction in cooperativity and a parallel increase in affinity during the analytical procedure in contrast to the original tonometric method. With the applied preparation technique, we could store the hemolysate for 70 days at -25 degrees C without any significant changes in the O2 binding properties (P < 0.05) demonstrating the high quality of this procedure for analysing fragile fish hemoglobins. The present investigation demonstrates that the oxygen affinity of the hemoglobins varied between the genotypes. At all temperatures, except 20 degrees C and pH 8.0, Hb-I(2/2) had a higher O2 affinity than Hb-I(1/1). These results conform with previous results (16), suggesting Hb-I(2/2), the genotype which is the dominant allele in northern areas, to be the most efficient O2 carrier at low temperatures. The highest O2 affinity, however, was found for Hb-I(2/2b), supporting the results of Fyhn et al. (9), that this genotype is more restricted to coastal and warmer water and thus a better marker of the coastal population. Our results further suggest a correlation between genotype specific growth rates and oxygen affinities at all temperatures studied, with the highest growth rates observed in those genotypes having the highest O2 affinities. In conclusion, the hemoglobin polymorphism of cod seems to be correlated with physiological performance.

Arctic adaptation in reindeer The energy saving of a hemoglobin

Previous results [(1988) Arct. Med. Res. 47, 83–88] have shown that hemoglobin from reindeer is characterized by a low overall heat of oxygenation. This particular aspect has been investigated further in a series of precise oxygen equilibrium experiments. The results obtained show a peculiar dependence of the temperature effect on the fractional saturation of hemoglobin with oxygen, which could be regarded as a very interesting case of molecular adaptation to extreme environmental conditions.

HEMOGLOBIN GENOTYPES IN TURBOT (SCOPHTHALMUS MAXIMUS RAFINESQUE), THEIR OXYGEN AFFINITY PROPERTIES AND RELATION WITH GROWTH

Abstract A growth trial lasting 15 months was conducted with 80 individually tagged turbot at four temperatures (10, 13, 16, and 19°C). Haemoglobins of the individually tagged fish were analysed by agar gel electrophoresis and the fish segregated into three genotypes named; Hb-I(1/1), Hb-I(1/2) and Hb-I(2/2). The genotype specific growth rates of the fish were analysed. In addition, the oxygen affinity properties of the hemoglobin genotypes were studied at 10 and 19°C. The genotype Hb-I(2/2) displayed the overall best growth at 10, 13 and 16°C, but no differences in growth among genotypes were seen at 19°C. Further, Hb-I(2/2) had the highest oxygen affinity at 10 and 19°C, whereas Hb-I(1/1) had the lowest oxygen affinity at both temperatures. In conclusion, the hemoglobin polymorphism in turbot seems to be correlated with physiological performance, and thus useful in future selection programs for turbot as a correlated trait with production characteristics.

Haemoglobin components and oxygen transport in relation to habitat distribution in triplefin fishes (Tripterygiidae)

Abstract Haemoglobin components were analysed for nine species of New Zealand triplefins and their isoelectric points (pI) ranged from 5.1 to 7.0. The number of well-expressed isohaemoglobins was larger in shallow-water and tidal pool species, ranging from four in Grahamina signata to eight in Grahamina capito, and were relatively cathodal. Two strongly anodal isohaemoglobins were expressed in the mid-depth species Ruanoho decemdigitatus and Ruanoho whero, and one in the deeper water species Karalepis stewarti and Forsterygion malcolmi. The red blood cell oxygen-binding properties were determined at 15 °C and 25 °C in the pH range 6.7–7.9 for the shallow-water species G. capito, the shallow to mid-depth species Forsterygion varium, and the deep-water species F. malcolmi. Oxygen affinity was highest for G. capito and the magnitude of the Bohr effect lower (Δlog P50/ΔpH = −0.37 at 25 °C, where P50 is the half-saturation coefficient) compared to the two Forsterygion species (Δlog P50/ΔpH = −0.52 to −0.59). Further, the cooperativity factor, n50, was lower in G. capito thus maintaining oxygen transport over a wide range of environmental oxygen pressures. Oxygen binding was similarly influenced by temperature in both G. capito and F. malcolmi (maximum heat of oxygenation ΔHmax = −27 kJ mol−1 and −37 kJ mol−1, respectively). Thus, triplefin fishes living in shallow, thermally unstable habitats possess a greater number of cathodally migrating isohaemoglobins, and their red blood cells have a higher oxygen affinity and reduced cooperativity which is less sensitive to changes in pH than do species occurring in more stable, deeper water habitats. Our analysis of an assemblage of closely related species circumvents some of the difficulties inherent in studies where interpretation of experimental results is confounded by phylogeny.

Thermodynamics of oxygen binding to arctic hemoglobins. The case of reindeer.

The most surprising characteristic of reindeer hemoglobin (Hb) concerns its response to changes in temperature. Thus, the shape of the oxygen-binding curve is strongly temperature dependent due to the difference in the enthalpy of oxygenation between the T and R state of the molecule. In fact, delta H of oxygen binding to the T state is strongly exothermic whereas that of the R state is very close to zero or possibly positive after correction for the heat of oxygen solubilization. Moreover, the allosteric transition T0----R0 has been found to display a negative delta H and a contemporaneous decrease in entropy, a behavior which is precisely the opposite of what has been reported for other hemoglobins. As a whole, reindeer Hb represents a beautiful example of the significance that comparative studies may have in assessing the general validity of the main properties of the hemoglobin molecule.

Fluctuating asymmetry and other parameters of morphological variation of eelpout Zoarces viviparus (Zoarcidae, Teleostei) from different parts of its distributional range

Five populations of eelpout Zoarces viviparus from the White Sea, the Baltic (Gulf of Gdansk and Gulf of Finland) and the North Sea (Wadden Sea and Hafrsfjord) were studied. Using 17 bilateral meristic characters (number of holes in cranial bones) we assessed the following parameters: (i) fluctuating asymmetry (minor deviations from perfect bilateral symmetry), (ii) the factorial component of total phenotypic variance, which is a measure of within-population heterogeneity and (iii) mean values of each character. Morphological similarity among samples was only weakly correlated with geographical distances between sample locations. The magnitude of fluctuating asymmetry was higher in the samples from the White Sea and Hafrsfjord than in samples from the Wadden Sea and the Baltic Sea. The sample from the Gulf of Gdansk was characterized by the lowest fluctuating asymmetry and the highest factorial variation. We propose that among the most important factors determining fluctuating asymmetry in eelpout populations are salinity and temperature conditions and the level of genetic variation.

THE PRIMARY STRUCTURE OF HEMOGLOBIN FROM REINDEER (RANGIFER TARANDUS TARANDUS) AND ITS FUNCTIONAL IMPLICATIONS

The primary structures of alpha- and beta-chains of hemoglobin from reindeer (Rangifer tarandus tarandus) were determined. Comparison of the reindeer hemoglobin sequence with those of human and bovine hemoglobins showed 50 and 29 substitutions per alpha beta dimer, respectively. The influence of replacements on the modulation of hemoglobin oxygen affinity by heterothopic ligands and temperature, as well as their importance on the structure-function relationships in hemoglobin are discussed.

Arctic life adaptation—II. the function of musk ox (Ovibos muschatos) hemoglobin

1. The hemoglobin system from musk ox (Ovibos muschatos) has been characterized from the functional point of view with special regard to the effect of organic phosphates and temperature. 2. The results are similar to those previously obtained in the case of reindeer and confirm that hemoglobins from arctic animals may display very low enthalpy change for the reaction with oxygen. 3. This finding is considered an example of molecular adaptation of respiratory pigments to extreme environmental conditions.