Swelling Of Red Cells Research Articles

Acute cerebrovascular effects in juvenile coho salmon exposed to roadway runoff

Urban stormwater is responsible for recurrent mortality events of coho salmon (Oncorhynchus kisutch) documented in the US Pacific Northwest. Currently, the toxic mode of action is unknown; however, mortality is often associated with large increases in hematocrit (Hct). We found that hemoconcentration and red cell swelling may offer a partial explanation for increased Hct, based on increased total hemoglobin and reduced mean cell hemoglobin concentration of the blood in runoff-exposed juvenile coho salmon, relative to controls. No significant changes were seen in plasma total thiols or antioxidant power, discounting osmoregulatory dysfunction and plasma dehydration as a driver of hemoconcentration. To test for plasma leakage, we injected Evans blue dye complexed with bovine serum albumin (EBD–BSA) intracardially and circulated for 5 min, followed by a 15 min washout with saline. Coho salmon exposed to roadway runoff showed significant accumulation of EBD–BSA in brain and olfactory rosette regions compared with controls, demonstrating plasma leakage from the cerebrovasculature. Our results suggest blood–brain barrier disruption underlies symptom development in coho salmon exposed to roadway runoff.

Effects of anaerobic exercise accompanying catch-and-release fishing on blood-oxygen affinity of the sandbar shark ( Carcharhinus plumbeus, Nardo)

Recovery from anaerobic exercise is thought to be prolonged in elasmobranchs because they lack several mechanisms for maintaining or increasing oxygen delivery that are present in teleosts. For example, teleosts increase hematocrit and maximal blood-oxygen carrying capacity through red cell ejection from the spleen. Teleosts also counteract the reduction in hemoglobin oxygen affinity resulting from metabolic acidosis through an adrenergic-mediated increase in red cell Na +–H + exchanger activity. To begin to assess the consequences of anaerobic exercise accompanying catch-and-release fishing occurring within the estuarine nursery habitats of juvenile sandbar sharks ( Carcharhinus plumbeus, Nardo), we constructed blood-oxygen equilibrium curves using samples from individuals 1 h after capture by hook and line (exercise-stressed) and samples from fully-recovered animals maintained in a shore-side tank (control sharks). We also compared exercise-stressed and control sharks for hemoglobin concentration, hematocrit, red cell count, intracellular pH, and nucleoside triphosphate concentration ([NTP]). In contrast to results from previous studies on elasmobranchs, we found an elevation in both hematocrit (≈ 21%) and blood hemoglobin concentration (≈ 10%) in exercise-stressed sharks. There was also clear evidence of red cell swelling. Mean red cell volume was ≈ 28% higher and mean cell hemoglobin concentration was ≈ 10% lower in exercise-stressed sharks. Most important, in spite of significant metabolic acidosis (0.3 pH units), blood from exercise-stressed sharks had an oxygen affinity equivalent to that of blood from control sharks. This was a direct consequence of intracellular pH being alkalinized by approximately 0.15 pH units relative to plasma pH in exercise-stressed sharks. Our results using isolated hemoglobin solutions showed that the observed reduction (≈ 15%) in intracellular [NTP] also contributed to the leftward shift in the oxygen equilibrium curves. As expected, we found sandbar shark red cells to be unresponsive to exogenous catecholamines. Regardless, sandbar sharks appear able to prevent the decrease in blood-oxygen affinity resulting from anaerobic exercise (and the concomitant decreases in plasma pH), as has been well-documented in teleosts. Our results suggest, therefore, that oxygen delivery following exhaustive exercise is not necessarily compromised in juvenile sandbar sharks, and that hook and line capture and subsequent release do not increase rates of mortality, although both are yet to be directly confirmed.

Physiological responses of the Asian sea bass, Lates calcarifer to water quality deterioration during simulated live transport: acidosis, red-cell swelling, and levels of ions and ammonia in the plasma

Water is a major expense when air-freighting live fish. However, reducing the volume of water relative to the amount of product usually results in more rapid water quality deterioration and higher mortality due to the higher concentration of excretory products in closed transport systems. The major water quality effects experienced by fish during transport are: low dissolved oxygen levels due to oxygen consumption by respiration; accumulation of carbon dioxide from respiration; depression of pH caused by carbon dioxide accumulation; and increased ammonia levels resulting from ammonia excretion. The physiological responses of seawater-adapted barramundi, Lates calcarifer, were studied during simulated live transport and transport under circumstances of elevated CO 2 or NH 3. Blood samples were removed from fish exposed to these treatments and compared to samples from control fish, (free in the tank) and fish confined in a box but with free-flowing seawater. Analysing the blood samples showed that simulated transport caused the plasma pH of the fish to fall, threatening the blood's ability to transport oxygen, but the red blood cells apparently defended their internal pH and oxygen transport capacity, and swelled measurably as a result. Exposing fish to unusually high carbon dioxide or ammonia levels caused plasma pH to fall to near lethal levels. The effects of both carbon dioxide and ammonia need to be considered when studying the responses of barramundi to live transport. Water quality parameters during fish transport do not act in isolation. Attempts to reduce carbon dioxide accumulation, for example, by using a buffer to control the water pH, may influence the fish's ability to excrete ammonia.

The effect of strenuous exercise and beta-adrenergic blockade on the visual performance of juvenile rainbow trout, Oncorhynchus mykiss.

It is hypothesised that the visual performance of rainbow trout, Oncorhynchus mykiss, will be impaired by strenuous exercise as a result of metabolic stress (blood lactacidosis) that activates the Root effect and limits the oxygen-carrying capacity of blood flowing to the eye. The ability to resolve high contrast objects on a moving background, as a measure of visual performance, was quantified pre- and post-exercise using the optomotor response. Strenuous exercise induced a metabolic acidosis (8.0 mmol l(-1) blood lactate) and a significant red cell swelling response but no change in the optomotor response threshold (120 min of arc) was observed. Beta-adrenergic blockade (propranolol) abolished post-exercise red cell swelling but optomotor response thresholds were still maintained at 120 min of arc despite a significant blood lactate load (7.8 mmol l(-1)). The choroid rete mirabile of the trout is extremely well developed (rete area:eye area = 0.39) and may maintain visual performance by ensuring a relatively direct supply of oxygen to the central regions of the avascular retina. Exercised fish under beta-adrenergic blockade exhibited an enhanced optomotor response at 240-300 min of arc. Assuming that these responses reflect "tunnel vision", adrenergic regulation of red cell function may preserve a high ocular PO(2) gradient that satisfies the oxygen demand of peripheral retinal cells.

Correlates of choroid rete development with the metabolic potential of various tropical reef fish and the effect of strenuous exercise on visual performance

It is hypothesised that the transport of oxygen to the retinal cells of fish with Root effect haemoglobins (Hb) is impaired by strenuous exercise due to a proton load that drastically reduces arterial haemoglobin–oxygen affinity. Routinely active reef fishes have enhanced oxygen transport and anaerobic (i.e. blood lactate loading) potentials relative to inactive species. Surprisingly, the development of the choroid rete mirabile (employed as an oxygen concentrating apparatus in the eye) is directly correlated with post-exercise lactate loads rather than with the magnitude of the Root effect and suggests that an increased development is adaptive for fish with high anaerobic potentials. The hypothesis that visual performance is reduced by strenuous exercise was tested in Lutjanus carponotatus using the optomotor response. Moderate blood lactate loads (2 mmol l −1 blood lactate) and red cell swelling responses were induced by exercise, but the optomotor response threshold (180 min of arc) was maintained. A moderate metabolic disturbance does not therefore appear to be a liability for the visual performance of a tropical fish in possession of Root effect haemoglobins.

Volume regulation by red blood cells from brown trout

Regulatory volume decrease, following physical swelling of red cells from brown trout Salmo trutta, was almost complete in oxygenated cells but much less in deoxygenated cells. There was a small, insignificant regulatory volume increase, following physical shrinkage. Amiloride had no effect on this response, indicating that hypertonic shrinkage did not activate the Na+/H+ exchanger. However, cell volume was increased markedly in shrunken cells by addition of noradrenaline, with deoxygenated cells showing complete recovery. These data show that the previously reported differences in volume regulation between the red cells of brown trout and rainbow trout Oncorhynchus mykiss are not present and that both species appear to have lost volume sensitivity of the Na+/H+ exchanger.

Volume regulation by red blood cells from brown trout

Regulatory volume decrease, following physical swelling of red cells from brown trout Salmo trutta, was almost complete in oxygenated cells but much less in deoxygenated cells. There was a small, insignificant regulatory volume increase, following physical shrinkage. Amiloride had no effect on this response, indicating that hypertonic shrinkage did not activate the Na+/H+ exchanger. However, cell volume was increased markedly in shrunken cells by addition of noradrenaline, with deoxygenated cells showing complete recovery. These data show that the previously reported differences in volume regulation between the red cells of brown trout and rainbow trout Oncorhynchus mykiss are not present and that both species appear to have lost volume sensitivity of the Na+/H+ exchanger.

Influence of D- and L-glucose on erythrocytes and blood viscosity.

Elevated blood glucose levels are associated with substantial morbidity and mortality. The pathomechanism behind it is not well understood. The aim of the present study was to investigate the effect of glucose on blood rheology. Blood from healthy volunteers was incubated with various concentrations of D- and L-glucose for 1 h at 37 degrees C. Whole blood viscosity at haematocrit 45% was measured at high and low shear rate (94.5 and 0.1 s(-1)). Erythrocyte shape and volume were assessed. Haemoglobin solutions were incubated with D-glucose for up to 96 h and the viscosity was measured. D-glucose dissolved in H2O and diluted with isotonic NaCl, added to whole blood (additional D-glucose concentrations 0-80 mM), led to a red cell swelling and an increase in blood viscosity at low shear rate (0.1 s(-1)). This process was reversible upon removal of D-glucose. L-glucose, which is not transported into the red cell by the D-glucose-specific transport protein GLUT-1, had no effect. When D-glucose was dissolved and diluted in autologous plasma, haematocrit and viscosity remained unaffected, but L-glucose decreased both values. Incubation of a haemoglobin solution with D-glucose at 37 degrees C led to a time-dependent increase in glycosylated haemoglobin (HbA1C) up to 8%, but left the viscosity unchanged. Blood glucose tested in a wide range of concentrations did not affect blood viscosity and morphological or biophysical properties of erythrocytes.

Integrated responses to exhaustive exercise and recovery in rainbow trout white muscle: acid-base, phosphogen, carbohydrate, lipid, ammonia, fluid volume and electrolyte metabolism.

White muscle and arterial blood plasma were sampled at rest and during 4 h of recovery from exhaustive exercise in rainbow trout. A compound respiratory and metabolic acidosis in the blood was accompanied by increases in plasma lactate (in excess of the metabolic acid load), pyruvate, glucose, ammonia and inorganic phosphate levels, large elevations in haemoglobin concentration and haematocrit, red cell swelling, increases in the levels of most plasma electrolytes, but no shift of fluid out of the extracellular fluid (ECF) into the intracellular fluid (ICF) of white muscle. The decrease in white muscle pHi was comparable to that in pHe; both recovered by 4 h. Creatine phosphate and ATP levels were both reduced by 40% after exercise, the former recovering within 0.25 h, whereas the latter remained depressed until 4 h. Changes in creatine concentration mirrored those in creatine phosphate, whereas changes in IMP and ammonia concentration mirrored those in ATP. White muscle glycogen concentration was reduced 90% primarily by conversion to lactate; recovery was slow, to only 40% of resting glycogen levels by 4 h. During this period, most of the lactate and metabolic acid were retained in white muscle and there was excellent conservation of carbohydrate, suggesting that in situ glycogenesis rather than oxidation was the major fate of lactate. The redox state ([NAD+]/[NADH]) of the muscle cytoplasm, estimated from ICF lactate and pyruvate levels and pHi, remained unchanged from resting levels, challenging the traditional view of the 'anaerobic' production of lactate. Furthermore, the membrane potential, estimated from levels of ICF and ECF electrolytes using the Goldman equation, remained unchanged throughout, challenging the view that white muscle becomes depolarized after exhaustive exercise. Indeed, ICF K+ concentration was elevated. Lactate was distributed well out of electrochemical equilibrium with either the membrane potential (Em) or the pHe-pHi difference, supporting the view that lactate is actively retained in white muscle. In contrast, H+ was actively extruded. Ammonia was distributed passively according to Em rather than pHe-pHi throughout recovery, providing a mechanism for retaining high ICF ammonia levels for adenylate resynthesis in situ. Although lipid is not traditionally considered to be a fuel for burst exercise, substantial decreases in free carnitine and elevations in acyl-carnitines and acetyl-CoA concentrations indicated an important contribution of fatty acid oxidation by white muscle during both exercise and recovery.

CARBONIC ANHYDRASE INJECTION PROVIDES EVIDENCE FOR THE ROLE OF BLOOD ACID-BASE STATUS IN STIMULATING VENTILATION AFTER EXHAUSTIVE EXERCISE IN RAINBOW TROUT

This study tests the hypothesis that the increase in blood PCO2 and associated respiratory acidosis after exhaustive exercise play an important role in stimulating ventilation during post-exercise recovery in fish. Injection of bovine carbonic anhydrase (10 mg kg-1) into the bloodstream of rainbow trout caused a persistent 40 % increase in the HCO3- dehydration capacity of the blood. The treatment was designed to increase CO2 excretion and therefore to reduce PCO2 build-up and acidosis after exercise. Aerobic and anaerobic swimming performance were not affected by carbonic anhydrase, and there were only very minor effects on arterial blood acid­base status in resting fish. However, carbonic anhydrase attenuated post-exercise increases in PaCO2 and decreases in pHa by about 50 % without altering arterial O2 variables, red cell swelling or the intracellular pH of the brain or muscle tissues. The effects on arterial pH (pHa) resulted largely from alleviation of the increase in PaCO2. In accordance with the original hypothesis, normal post-exercise hyperventilation was greatly attenuated, through reductions in both ventilatory stroke volume and frequency, and excess post-exercise O2 consumption was reduced. Post-exercise increases in plasma levels of adrenaline and noradrenaline were also reduced by the carbonic anhydrase treatment. Overall, there was a strong correlation between increases in relative ventilation and decreases in pHa after exhaustive exercise. The results provide functional significance for the phenomenon of PaCO2 elevation and associated respiratory acidosis after exercise and are consistent with other recent studies indicating an important secondary drive to ventilation in fish based on arterial acid­base status, in addition to the primary drive based on arterial O2 levels.

How does environmental pollution affect red cell function in fish?

The present article reviews the effects of environmental pollution on ion transport, metabolism and shape of teleost red cells. Catecholamines liberated into the blood stream in acute stresses activate sodium/proton exchange across the red cell membrane causing red cell swelling. The exchanger is inhibited by alkyltin compounds and by methaemoglobinaemia. Amphiphiles increase the potassium permeability of the red cell membrane. Exposure to lead inhibits and exposure to tetrachloro-1,2-benzoquinone activates red cell δ-aminolevulinic acid dehydratase. Resin acids, cadmium, lead and oxidizing agents such as monochloramine induce the appearance of abnormal cell shapes. In addition, environmental contamination is often accompanied by an increased frequency of micronuclei in the red cells. Specifically, this effect has been observed during exposure to benzo(a)pyrene. These, and other, examples show that many red cell parameters respond to environmental pollution. Thus, determinations of red cell ion concentrations, microscopic examination of red cells and assays of red cell enzyme activities could be a useful addition to conventional pollution monitoring studies.

Species differences in the adrenergic responses of fish red cells: studies on whitefish, pikeperch, trout and carp

The occurrence and pH dependence (pHe 7-8) of the adrenergic red cell responses of two salmonids, trout and whitefish, and a percinid, pikeperch were studied. These are all species that live in well-oxygenated waters. The responses were compared to those of carp, which tolerates oxygen-deficient waters.The adrenergic responses of trout and whitefish red cells were pronounced. In these species red cell swelling, the accumulation of sodium and chloride in the cell, and the increase in red cell oxygen content at atmospheric oxygen tension were maximal at pH 7.3. In contrast, pikeperch red cells responded to β-adrenergic stimulation only at extracellular pH 7.1. In carp, the adrenergic response, occurring below extracellular pH 7.5, was small as compared to the two salmonids. In each case the onset of the adrenergic response coincided with the onset of the Root effect.The differences in the adrenergic responses between the two salmonids and pikeperch suggest that the occurrence of the adrenergic response is not directly related to the environmental oxygen requirements of the species, but may be linked to the activity pattern.

Physiological disturbances in rainbow trout (Salmo gairdneri) during acid and aluminum exposures in soft water of two calcium concentrations

Rainbow trout (Salmo gairdneri) fitted with dorsal aortic cannulae were exposed in a flow-through soft water system to three acidities (pH 5.2, 4.8, or 4.4) and two concentrations of Ca (45 or 410 μequiv.∙L−1), in the presence (105 μg∙L−1) or absence of Al. Blood was sampled for respiratory gases, ions, metabolites, and hematology before and at 4, 18, 28, 42, and 66 h exposure. Two toxic mechanisms of Al and acidity were seen: (i) ionoregulatory toxicity, which was caused by Al at pH 5.2 and 4.8 and by acidity at pH 4.4, and (ii) respiratory toxicity, which was caused solely by Al, and was greatest at higher pH. Ionoregulatory toxicity involved decreases in plasma Na+ and Cl−, red cell swelling, and hemoconcentration. Respiratory toxicity involved reduced blood oxygen tension, elevated blood carbon dioxide tension, and increases in blood lactate. Blood acidosis was a combination of respiratory acidosis (due to CO2 accumulation in the blood; higher pH exposures) and metabolic acidosis (probably due to differential Na+ and Cl− loss into the external, acidic environment; lower pH exposures). Higher water Ca reduced ionoregulatory disturbances due to acidity alone but not those due to Al at higher pH. Higher water Ca also reduced respiratory disturbances at lower pH but not at higher pH. The results are discussed with reference to the chemistry of Al and changes in the gill epithelium associated with acid and Al exposure.

Use of filtration methods in evaluation of the condition of fish red blood cells.

Use of filtration methods in evaluation of the condition of fish red blood cells.

Volume control of erythrocytes during storage. The role of mannitol.

When red cells are stored as suspensions in artificial preservative solutions such as those consisting of saline, adenine, and glucose, increased hemolysis of the red cells occurs in vitro, a phenomenon that is readily prevented by the addition of mannitol. The mechanism by which mannitol prevents hemolysis is unknown. The authors have examined the possibility that mannitol prevents hemolysis by prevention of osmotic swelling of red cells that might otherwise increase their volume beyond their critical hemolytic volume. Storage in artificial media does, indeed, result in swelling of the red cells, which is prevented by mannitol and is increased by the addition of nystatin, an agent that greatly increases the sodium-potassium permeability of the cells. Swelling and hemolysis are correlated at different concentrations of mannitol, but the correlation breaks down when the case of nystatin-treated cells is considered; even with marked swelling, there is relatively little hemolysis. Moreover, increasing the volume of the stored red cells acutely by osmotic means results in relatively little additional hemolysis. These findings make it appear unlikely that simple osmotic lysis is the cause of hemolysis in stored cells. Some other stabilizing effect of mannitol that may or may not be related to its osmotic effect must play a role.

Effect of adrenaline and blood gas conditions on red cell volume and intra-erythrocytic electrolytes in the carp, Cyprinus carpio.

1. Carp, when subjected to air stress in vivo, developed a hypoxaemia associated with an acidosis due to elevation of PCO2 and arterial lactate content. Adrenaline and noradrenaline levels rose markedly, and the mean corpuscular haemoglobin concentration (MCHC in mmol Hb l-1 red blood cells) decreased, indicating swelling of the erythrocytes. 2. No effect of adrenaline could be observed in vitro after equilibration at normal pH and oxygenation level. 3. Simulation of air stress in vitro, by using hypercapnic hypoxia together with adrenaline, caused appreciable swelling of the red cells. The effect of adrenaline could be blocked by propranolol and was reversible when the blood gases were readjusted to normal values. Swelling due to hypercapnic hypoxia was not affected by propranolol. 4. At normal PO2, lowering pH by increasing PCO2 or adding HCl to the blood at PCO2 = 3.5 mmHg (1 mmHg = 133.3 Pa) both led to swelling of red cells; further swelling occurred when adrenaline was added. 5. At normal or even elevated intracellular pH, hypoxia led to red cell swelling; further swelling occurred when adrenaline was added. 6. The swelling of red cells under hypoxic and/or acidotic conditions (induced by either added HCl or increased PCO2) was associated with an increase in the intracellular K+, Na+ and Cl- levels, together with an increase in the Donnan distribution ratio of Cl-, rCl. delta K+:delta Na+ was 4:1. The swelling associated with the addition of adrenaline was also accompanied by an increase in K+, Na+, Cl- and rCl. In this case the ratio delta K+:delta Na+ was 1:2.4. 7. There was a correlation between the Donnan ratio, rCl, and the change in MCHC, even in those cases where extracellular and intracellular pH were above normal. In addition to pH and oxygen saturation, rCl should therefore be considered as a possible triggering factor for the action of adrenaline. 8. In addition to the increased catecholamine levels, stress during sampling may be inferred if low values of MCHC and high values of intraerythrocytic chloride and a high chloride distribution ratio, rCl, are found in whole blood.

The effects of acid and acid/aluminum exposure on circulating plasma cortisol levels and other blood parameters in the rainbow trout, Salmo gairdneri

Softwater (Ca2+=50, Na+= 50(μequiv. l−1) acclimated rainbow trout were fitted with chronic arterial catheters to allow for repetitive blood sampling. After 48 h recovery they were then exposed to either control (pH 6.5, Al = 0μg l−1), acid (pH 4.8, Al = 0μg l−1) or acid plus aluminum (pH 4.8, A1 = 112 μg l−1) conditions for 72 h. Parameters measured included blood glucose, lactate, haemoglobin, haematocrit and plasma Na+, Cl−, protein and cortisol.Exposure to pH 4′8 alone caused no mortality, a moderate ionoregulatory disturbance and a transient elevation in plasma cortisol. All other parameters were not significantly different from controls. Addition of aluminum to this exposure caused 100% mortality with a mean survival time of only 27.0 h. There was a marked decrease in plasma ions, hyperglycemia, lactate accumulation, haemoconcentration, red cell swelling, and a sharp rise in plasma cortisol becoming greatly increased as the fish neared death. The mechanism of toxicity of acute acid/aluminum exposure, the role for cortisol under such conditions, and the validity of cortisol and glucose as indicators of stress in fish are discussed.

Effects of adrenaline on ionic equilibria in red blood cells of rainbow trout (Salmo gairdneri)

Effects of adrenaline on the equilibrium distributions of Na(+) , K(+) , H(+) , Cl(-) , and H2O across the cell membrane of rainbow trout (Salmo gairdneri) erythrocytes were determinedin vitro, as a function of P CO2 (1.76-7.77 torr). CO2-carrying capacity of the blood was also examined. Plasma catecholamine concentrations inunanaesthetized, unrestrained trout were 3.1 nM adrenaline and 1.2 nM noradrenaline. Elevation of the plasma adrenaline concentrationin vitro to 4.6 × 10(3) nM resulted in net gains of Na(+) , Cl(-) and H2O by red cells, a net loss of H(+) from red cells, and a pronounced red cell swelling. Adrenaline also reduced the CO2-carrying capacity of trout bloodin vitro. The magnitudes of these effects increased with PCO2 and, thus, were sensitive to blood HCO3 (-) concentrations. The distribution of K(+) between red cells and plasma was unaffected by adrenaline. Adrenergic-mediated ion movements and red cell swelling were sensitive to both propranolol and SITS. These results are consistent with the symport NaCl uptake model for adrenergic-mediated swelling of Baroinet al. (1984). The adrenergic response of fish erythrocytes may function to ameliorate the effects of blood acidoses on O2-carrying capacity by maintaining red cell pH in the face of a decrease in plasma pH.

Influences of exercise-stress and adrenaline upon intra- and extracellular acid-base status, electrolyte composition and respiratory properties of blood in tench (Tinca tinca) at different seasons.

Exercise-stress in tench resulted in severe acidosis in both the red cells and the extracellular fluid in vivo. These coincident pH decreases conformed to the in vitro pHi-pHe relationship for tench blood in the oxygenated state. The extracellular acidosis was primarily respiratory in winter and metabolic in spring and summer. This was due to more effective buffering of metabolic protons in winter by an elevation in [HCO3-] levels, rather than to differences in the lactic- and carbonic acid loads. A good correspondence was found between buffered metabolic protons and increases in [lactate]. There was no evidence for beta-adrenergic red cell swelling and associated red cell pH changes in tench both after exercise and adrenaline infusion. Arterial O2 transport was, however, improved in exercise by pronounced increases in PO2. This increase was not of adrenergic origin, as adrenaline alone caused a decrease in arterial PO2. Large increases in plasma potassium concentration and small elevations of chloride and calcium levels occurred in exercise. Hematocrit and blood [Hb] also increased, probably due to an adrenergic release of erythrocytes from the spleen, but these increases were small and appeared unimportant for blood O2 transport. Seasonal differences were found in exercise-induced changes in [lactate], in the magnitude of electrolyte and PO2 changes, as well as in resting values for pHe, pHi, [HCO3-], [Cl-] and [Ca++]. The origin and importance of these are discussed.

Blood oxygen transport and acid-base status of stressed trout ( Salmo gairdnerii): Pre- and postbranchial values in winter fish

1. 1. Rainbow trout, obtained from a temperature below 5°C and kept at 13–14°C for 2 weeks, were cannulated via both dorsal and ventral aorta, and chased for 5 min at 14°C. 2. 2. The stress caused a marked extracellular acidosis which had both respiratory and metabolic components. 3. 3. The dorsal aortic red cell pH also decreased, from 7.33 to 7.12, as a result of the stress. 4. 4. This caused a drop in the arterial oxygen saturation, although arterial oxygen tension remained unchanged. 5. 5. Additionally, no red cell swelling was observed. Also, the carbon dioxide and red cell chloride data suggest that bicarbonate influx into the red cells and, consequently, carbon dioxide excretion increased in stress. 6. 6. These findings strongly suggest that the red cells of the winter fish used lacked beta-adrenergic responses. 7. 7. In the apparent absence of beta-adrenergic responses, the ventral aortic red cell pH was maintained at a nearly constant level, probably by the Haldane proton uptake upon deoxygenation of the blood in the tissues. 8. 8. Consequently, and since the ventral aortic oxygen tension remained unchanged, the oxygen saturation of venous blood was not affected by stress.