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1. Relative to human HbA, opossum (Didelphis virginiana) hemoglobin was found to be more susceptible to autoxidation. While the initial rate of autoxidation of spot (Leiostomus xanthurus) hemoglobin is close to that of HbA, complete oxidation occurs in 50 hr. 2. Direct addition of hydrogen peroxide (H2O2) induced oxidation of hemoglobins in a definite order: spot Hb > HbA > opossum Hb. Excess H2O2 led to heme degradation and precipitation that occurred much faster for spot Hb than the case with other proteins. 3. Exposure of hemoglobins to a continuous flux of H2O2, generated by the glucose/glucose oxidase system, induced the formation of heterogeneous protein-associated oxidation products. 4. Differential reactivity among these hemoglobins under the same or different oxidative conditions, with respect to methemoglobin formation and stability of the ferric form, may reflect the differences in the local heme environment of these proteins.

1. Diet, time from last feeding, temperature, season and sexual stage are some of the factors influencing the lipoprotein pattern. 2. Keeping these factors constant species-specific differences observed among lipoprotein patterns of Sparus aurata, Puntazzo puntazzo, Diplodus sargus, Diplodus vulgaris and Dicentrarchus labrax are discussed. 3. Feeding habits and therefore lipid absorption and the rate of lipoprotein maturation process are the factors determining the observed differences.

1. The blue-green coloration of the blood plasma in some marine fishes, which is attributed to a protein bound tetrapyrrol (biliverdin), is an anomaly in vertebrates. 2. Recent studies have shown that biliverdin not only occurs in many fish, but is also present in the blood of tobacco hornworm, the wings of moth and butterfly, the shell of bird eggs, the serum and egg of frog, the placenta of dog and in the blood of humans suffering from hepatic diseases. 3. In this review, we begin with a historical account of the description of the presence of blue-green blood plasma in fish, and then consider the biochemistry, metabolism, physiology, and the ecological implications of biliverdin in fish. 4. A comparative description of the occurrence of biliverdin in fish and other animals is presented. 5. The mechanism of accumulation of biliverdin in fish blood and its evolutionary significance are also considered. It is suggested that this process may serve as a useful model for further research on bile pigment metabolism in other animals.

1. Notothenia coriiceps neglecta is a cold-adapted notothenioid teleost, widely distributed in the Antarctic waters. 2. In comparison with fishes from temperate waters, the blood of this teleost contains a reduced number of erythrocytes and concentration of hemoglobin; the erythrocytes contain two hemoglobins, Hb1 and Hb2, respectively accounting for approximately 90, and 5% of the total. 3. The two components differ by the alpha chain; the amino acid sequence of the beta chain in common to the two hemoglobins has been established, thus completing the elucidation of the primary structure of the major component Hb 1.

1. Serum proteins of upstream migrants of Petromyzon marinus were separated electrophoretically on polyacrylamide and agarose as well as by crossed immunoelectrophoresis using anti-lamprey serum antiserum. 2. Some constituents of four of the five major bands separated by electrophoresis on polyacrylamide were located in the agarose and crossed-immunoelectrophoretic patterns. 3. Two serum proteins, SDS-1 and CB-III, were isolated. SDS-1 was found to be a glycoprotein; CB-III may be a lipoprotein. 4. Monospecific antisera were produced against SDS-1 and CB-III and were used in crossed immunoelectrophoresis to analyze lamprey serum. This method revealed that electrophoretically different forms of these two proteins exist in serum. 5. Immunoelectrophoretic methods demonstrated that three of the major bands produced by polyacrylamide gel electrophoresis of whole serum contained more than a single protein.

1. The half-saturation value, P50, for 'stripped' hemoglobins from Trematomus spp. (fam. Nototheniidae) at pH 8.27 and -1.8 degrees C varied from 12.1 mmHg in the pelagic species T. borchgrevinki to 1.3 mmHg in the sedentary benthic species T. centronotus. 2. The nototheniid hemoglobins showed appreciable Bohr effects (phi = delta log P50/delta pH = -0.87 to -0.48) while the hemoglobin from a bathydraconid species, Gymnodraco acuticeps, was insensitive to pH; phi = -0.02 No Root shifts were detected. 3. Cooperative oxygen binding was present in all hemoglobins with Hill's coefficient, n, ranging from 1.21 +/- 0.14 in Dissostichus mawsoni to 2.28 +/- 0.88 in T. borchgrevinki among the Nototheniidae, and n = 1.50 +/- 0.14 in G. acuticeps. 4. A small increase in P50- was promoted by ATP for hemoglobins from T. borchgrevinki, T. bernacchii and D. mawsoni but significant temperature effects on oxygen binding were manifested, with apparent heats of oxygenation, delta H, = -56.13, 62.16, and -23.98 kJ mol-1, respectively.

Red blood cell soluble phosphates from Semaprochilodus taeniurus, S. insignis and a possible hybrid jaraqui-açu, from Amazon basin, were studied by column chromatography. The Semaprochilodus school and the jaraqui-açu shows an intermediate percentage of total phosphorus content as GTP but not as ATP. GTP/ATP ratio for jaraqui-açu and S. taeniurus were 1.5 and for S. insignis, 3.0.

The ligand binding properties of the Root effect haemoglobin of the marlin have been investigated in the temperature range 12-35 degrees C. An essentially symmetric displacement of the binding isotherms to higher concentration is observed on raising the temperature. Thermodynamic parameters have been obtained for the equilibrium binding constants, in terms of the two state model for co-operativity. Kinetic measurements indicate chain heterogeneity in both the T and R states. Activation parameters have been obtained for both chains in both quaternary states.

1. The purine nucleotides were determined in the whole blood of 9 fresh water teleosts and 2 marine selachians. 2. GTP and ATP accounted for 88-99% of the total erythrocytes purines. 3. The ATP/ADP ratio ranged from 11 to 60 in the erythrocytes of the fish examined. 4. GTP is widely distributed in fish erythrocytes but its level ranged from 1 to 33 nmol/mg Hb (0.4 to 9 mumol/ml erythrocyte). 5. Lepomis and Esox exhibited a GTP/ATP ratio as elevated as in Anguilla; moreover the concentration of GTP per mol of Hb (physiologically most indicative) is higher in Lepomis, Esox, Ictalurus and Silurus than in Anguilla.

1. The myogen protein patterns of lateral muscle of Liza ramada (Risso) and Chelon labrosus (Risso), revealed by isoelectric focusing, are reported. Specific differences are noted both in the white and in the red muscle (where they are more evident). 2. Red muscle shows the presence of a chromoprotein, found to be myoglobin, with a pI characteristic for the species. 3. Blood haemoglobins were examined with the same technique and also found to be species specific.

1. The initial rate of cholesterol esterification and the concentration of triacylglycerols in plasma were generally lower in prespawning than in immature chars. No typical pattern of variation of plasma cholesterol could be observed. 2. The observed variations in plasma cholesterol esterification and lipids may be caused by reduced feeding in the prespawning period. 3. The initial rate of cholesterol esterification in plasma was positively correlated with the concentrations of triacylglycerols and unesterified cholesterol in plasma, respectively.

Vitellogenin was isolated by affinity chromatography and gel filtration from landlocked Atlantic salmon plasma. Vitellogenin was labelled with iodine-131 using iodogen and an homologous radioimmunoassay was developed. There was poor immunological cross-reactivity with vitellogenin or plasma from other teleosts. Parallelism of the vitellogenin standard to the displacement by plasma of vitellogenic salmon allowed the assay to be used to evaluate the seasonal concentration profile of vitellogenin in female adult salmon. Extracts of liver or ovary from female Atlantic salmon also yielded displacements parallel to the vitellogenin standard in the assay.

Hemoglobin electrophoretic pattern, whole blood Bohr effect, intraerythrocytic phosphates and some hematological parameters were determined and compared between lotic and lentic Hypostomus regani specimens. The electrophoretic patterns of specimens from both kinds of environments showed four anodic fractions. The relative concentration of the fraction I (the most anodal) is significantly different between specimens from lotic and lentic ambients. Both lentic and lotic specimens presented a low Bohr effect: phi = -0.012 to lentic habitat and phi = -0.187 to lotic one. ATP/GTP ratio are higher than 1.0 for lotic environment specimens and lower than 1.0 for lentic ones.

1. The purine nucleotides were determined in the whole blood of 13 species of seawater teleosts and 3 species of selachians. 2. The ATP/ADP ratio ranged from 4 to 50 in the erythrocytes of the fish examined. 3. GTP is widely distributed in fish erythrocytes but its level ranged from 2 to 27 nmol/mg Hb, the highest value being observed in Conger. 4. Conger, Melanogrammus and Merlangus exhibited a GTP/ATP ratio higher than 1. The concentration of GTP per mol of Hb (physiologically most indicative) is higher than 1 in Conger, Melanogrammus, Merlangus and Pollachus, and close to 1 in Molva and Gadus.

1. Prostaglandin (PG) syntheses from labelled highly unsaturated fatty acids were investigated in washed thrombocyte suspensions of four species of marine fish, flounder (Paralichthys olivaceus), black seabream (Acanthopagrus schlegeli), black rockfish (Sebastes schlegeli), and red seabream (Pagrus major). 2. Synthesized PGs were analyzed by thin-layer radiochromatogram scanner and high-performance liquid chromatography. 3. When [1-(14)C]arachidonic acid (AA) was incubated with washer thrombocyte suspension, AA was mainly converted to PGF(2alpha), PGE2, and PGD2 in all species. 4. It was suggested that [1-(14)C]eicosapentaenoic acid was mainly converted in PGs only in black rockfish thrombocytes, and the metabolites were mainly PGE3 and PGD3. 5 However, [1-(14)C]docosahexaenoic acid was not cyclized into PGs in all the species.

1. The hemoglobin of the sting-ray, Dasyatis sabina, is both polymorphic and heterogeneous; three components predominate. 2. One major component has two kinds of polypeptide chain, of which one, presumably an alpha-chain, has a blocked NH2-terminus and an arginyl COOH-terminus, whereas carboxypeptidases A and B release tyrosine and histidine from the COOH-terminus of the beta-chain. 3. The amino acid sequence of the beginning NH2-terminal segment of the beta-chain of the major component has been determined. 4. The hemoglobin of the sting-ray, Dasyatis sabina, is highly resistant to urea and does not dissociate readily into subunits. 5. Oxygen binding by the hemoglobin is not affected by organic phosphates or high concentrations of either NaCl or urea. 6. The hemoglobin does not polymerize beyond tetramers. 7. Cooperativity, as monitored by n in the Hill equation, is pH-dependent and maximal between pH 8.5 and 9.0. 8. The hemoglobin has a large Bohr effect; the oxygen affinity is 16 times higher at pH 10 than at pH 6.5.

The physiological adaptations of skipjack tuna lead to 10 degrees C changes in blood temperature every few seconds. The change in reaction rate of red blood cell LDH, MDH and GAPD to change in temperature are large under saturating substrate conditions but under intra-cellular substrate conditions reaction rate is almost independent of temperature. It seems likely that the biochemical effects observed are mediated through changes in Km with temperature. Three species related to the skipjack, but lacking its extreme physiological adaptations, had many of the biochemical adaptations seen in the skipjack. The primitive ancestor of the skipjack probably had the biochemical preadaptations needed to allow the physiological evolution seen in this lineage.

Electrophoretic patterns of hemoglobins, red blood cell phosphates, Bohr effect of blood and stripped hemoglobin, ATP and GTP modulation of Hb-O2 affinity of two species belonging to the genus Semaprochilodus (S. insignis and S. taeniurus--jaraquis), captured at two different locations in Rio Negro, near Manaus (Amazonas), are described. High Bohr effect values were found in these two species and in jaraqui-açu (a possible hybrid between them). In S. insignis a higher Bohr effect for migrating individuals was observed, while S. taeniurus showed the reverse. The main erythrocytic phosphates found in these two species are ATP and GTP. They have a large influence on the O2-Hb binding properties in both species. The GTP/ATP ratio showed large differences between catching places. The two species have different electrophoretic patterns. Semaprochilodus insignis showed four anodal components which showed quantitative variation between catching places; S. taeniurus had three anodal components and a similar quantitative variation.

Basic composition and properties of isolated transferrins of Silurus glanis and Esox lucius have been compared. In transferrin of S. glanis carbohydrate is absent, but it is present in transferrin of E. lucius (2.5%). The N-terminal amino acid is alanine in both species. Mol. wts are 68,400 (S. glanis) and 86,800 (E. lucius). Transferrins of the two species are heterogeneous, but genetic polymorphism was not observed.

Total hemolysates of Synbranchus marmoratus Bloch, 1795 captured at four different sites in the State of São Paulo, Brazil, showed two different hemoglobin phenotypes when submitted to agar-starch gel electrophoresis on glass slides in basic buffer. Phenotype I was characterized by 3 hemoglobin bands. When the total hemolysate was submitted to cellulose acetate electrophoresis in basic buffer containing 6 M urea and beta-mercaptoethanol, Phenotype I showed four globins of the alpha 1, alpha 2, beta and gamma types, with 11.9 +/- 1.9 g% total hemoglobin, 45.3 +/- 3.6% globular volume, and 26.8 +/- 4.4% mean corpuscular hemoglobin concentration (MCHC). Phenotype II showed three groups of hemoglobins, with a total of up to 12 hemoglobin bands. When the total hemolysate was submitted to cellulose acetate electrophoresis in basic buffer containing 6 M urea and beta-mercaptoethanol, phenotype II showed five types of globins, denoted types alpha 1, alpha 2, gamma 1, gamma 2 and beta, having electrophoretic positions different from those of Phenotype I globins, with 18.1 +/- 3.3% total hemoglobin, 47.9 +/- 6.4% globular volume, and 37.8 +/- 4.4% MCHC. The distribution of the specimens having the two hemoglobin phenotypes is associated with the different geomorphological provinces of the State of São Paulo, suggesting the existence of at least two populational groups of Synbranchus marmoratus.

Total hemolysates of Synbranchus marmoratus Bloch, 1795, captured in Vitoriana, district of Botucatu, State of São Paulo, Brazil, were submitted to agar-starch gel electrophoresis on glass slides using 42 mM-Tris 1.7 mM EDTA-6.1 mM borate buffer, pH 8.8, for the gel and 10 mM borate-1.7 mM NaOH buffer, pH 8.6, for the cuvette. Three distinct hemoglobin bands were detected, with Hb I being of the cathodic type. Cellulose acetate electrophoresis in 800 mM Tris-2.1 mM EDTA buffer, pH 8.9, containing 6 M urea and 2.25 mM beta-mercaptoethanol indicated the presence of four globin chains denoted alpha 1, alpha 2, beta and gamma. It is suggested that the probable tetrameric constitution of the hemoglobin of Synbranchus marmoratus Bloch, 1795 is Hb I (alpha (2)2 gamma 2), Hb II (alpha (1)2 gamma 2) and Hb III (alpha (1)2 beta 2).

Plasma corticosteroids of eighteen vertebrate species were separated by reversed-phase high-performance liquid chromatography (HPLC). The major corticosteroids identified in these species by HPLC agreed with previous reports which used vigorous criteria for identification. The HPLC technique was validated for the assay of cortisol in mullet plasma and corticosterone in the rat. Qualitative as well as quantitative changes in corticosteroid secretion were detected by HPLC in several vertebrates during exposure to stress.

Five isohemoglobins were characterized from the marine teleost fish, Girella tricuspidata, using polyacrylamide gel electrophoresis and isoelectric focusing. O2-equilibrium (OEC) studies of the five components separated by column chromatography showed functional heterogeneity with respect to Hill's coefficient n, the O2 affinity, P50, and the Bohr coefficient phi, = delta logp50/delta pH. Recombination and examination of the isolates gave an OEC similar to that of "stripped" hemolysate and similar to a curve synthesized from a model incorporating the OEC parameters of the isoHbs.

1. Vitellogenin was purified from plasma of estrogen-treated spotted seatrout (Cynoscion nebulosus) and was shown to be a protein of approximately 200,000 mol. wt. 2. Antisera were raised against the isolated protein and were used to develop a radioimmunoassay for spotted seatrout vitellogenin. 3. The substance measured was shown to be female-specific, hepatically-derived, estrogen-inducible and immunologically related to oocyte yolk proteins. 4. Vitellogenin levels in the plasma of female spotted seatrout were shown to rise dramatically during the period of ovarian recrudescence.

1. Mean monthly serum levels of total protein, C-reactive protein (CRP) and serum amyloid P component (SAP) in plaice, showed no significant difference between the sexes. 2. Highest values for CRP and protein were found between June and September, with no significant seasonal variation in SAP. 3. There was no change in CRP concentration in plaice maintained for 7 days at a higher temperature of 18.5 degrees C. 4. Injection of lipopolysaccharide caused the highest value for CRP on day 1 and for the spleen index on day 5 after injection. 5. Phagocytic stimulation with carbon had no significant effect on the CRP response to endotoxin.

The binding characteristics of biliverdin to its carrier protein in the blue-green blood of Clinocottus analis (woolly sculpin) and Anguilla japonica (freshwater eel) were operationally defined by various chemical probing methods and spectra analysis. The biliverdin in C. analis is a rotatable coiled molecule enveloped in a hydrophobic pocket of its carrier protein. The biliverdin in A. japonica is an open form molecule with external hydrogen bond or weak ester bond interacting with the carrier protein. This, for the first time, shows that the novel firm binding biliprotein complexes in the blood of different species are biochemically different. Their comparative and evolutionary significance is discussed.

Considering the presently available data it is clear that the Root effect represents an exaggerated alkaline Bohr effect which occurs in the absence of a normal acid Bohr effect and is associated with a loss of oxygen binding capacity at low pH. Undoubtedly at the molecular level the presence of a Ser residue at position F9(94) beta in these haemoglobin is of primary importance. No Root effect haemoglobin has yet been identified which lacks this substitution. On the other hand however many haemoglobins are known which possess this Ser residue and at the same time lack a Root effect. Other factors arising from interactions at other sites in the haemoglobin molecule are obviously sufficient to negate the otherwise stabilizing effect of this critical Ser residue. The loss of cooperativity of Root effect systems as the pH is lowered is readily explained as due to stabilization of the low affinity T state to such a degree that the switch to the high affinity R state is suppressed even in the fully liganded molecule. The observation of Hill coefficients of less than unity requires that within the T state chain heterogeneity exists such that the alpha and beta chain haems demonstrate significantly different affinities for ligand. The physiological role of Root effect haemoglobins is demonstrably not inevitably linked to the swim bladder but more probably arose from the need to oxygenate the poorly vascularized retina of many fishes.(ABSTRACT TRUNCATED AT 250 WORDS)

1. The plasma membrane of the flounder erythrocyte contains a Mg2+-dependent ATPase which is insensitive to ouabain. Mg2+ is part of the substrate, Mg-ATP, and Mg2+ also functions as a nonessential activator. 2. Ca2+, Mn2+ and Co2+ can replace Mg2+ as an activator of ATP hydrolysis. Cu2+ and Zn2+ abolish the Mg-dependent activity. It is shown that Ca2+ and Mg2+ activate the same enzyme and that Mg-ATP and Ca-ATP are mutually competitive. 3. The hydrolysis of ATP obeys Michaelis-Menten kinetics whether or not the Mg2+-ATPase is fully activated by Mg2+. The KM values for Mg-ATP were found to be 0.13 and 0.43 mM respectively. 4. Free ATP acts as a competitive inhibitor towards Mg-ATP and the dissociation constant for the enzyme-ATP complex was determined to be about 0.55 mM. 5. The Mg2+ -ATPase has a low specificity and reacts with the common nucleoside triphosphates GTP, ITP, UTP and CTP. 6. The enzyme has a broad pH optimum ranging from 6.5 to 7.2 and an energy of activation of 13.5 kcal/mol between 0 and 30 degrees C. 7. The effect of some activators and inhibitors of membrane-bound ATPases are reported.

NADH-methemoglobin reductase activity of erythrocytes from the coho salmon, Oncorhynchus kisutch, sockeye salmon, Oncorhynchus nerka, and the rainbow trout, Salmo gairdneri exhibited a major band of activity that resembled the human enzyme in electrophoretic mobility. No polymorphism was found in 35 samples from rainbow trout, 4 samples from Dolly Varden, 29 samples from sockeye salmon, and 24 samples from coho salmon. All samples differed from the human enzyme in that they appeared to be membrane-bound and required the presence of a detergent, Triton X-100, for solubilization. Rainbow trout and coho salmon enzymatic activity is greater than the human enzyme activity at 15 degrees C.

Channel catfish were acclimated in vivo to 12, 17, 22 or 27 degrees C and their peripheral blood erythrocytes, thrombocytes, T lymphocytes and B lymphocytes assayed for cellular fatty acid composition. Excepting cells from 12 degrees C acclimated fish, all cells responded to acclimation to lower temperatures by exhibiting increased levels of phospholipid unsaturated fatty acids. Although temperature independent differences were observed between erythrocytes, thrombocytes and lymphocytes, no differences between T lymphocytes and B lymphocytes were seen.