SUMMARY Although the retina has one of the highest metabolic rates among tissues,certain teleost fishes lack any vascular supply to this organ which, in combination with the overall thickness of the organ, results in extremely long diffusion distances. As the only way to compensate for these obstacles, oxygen partial pressure ( P O 2 ) in the eyes of such fish is elevated far above atmospheric values. Although not supported by any direct evidence, the enhancement of P O 2 is considered to be related to the Root effect, the release upon acidification of Hb-bound O 2 into physical dissolution, possibly supported by counter-current multiplication similar to the loop of Henle. The present study evaluates the magnitude of intraocular P O 2 enhancement under tightly controlled physiological conditions, to directly confirm the involvement of the Root effect on intraocular P O 2 in the retina of rainbow trout Oncorhynchus mykiss . Intraocular P O 2 was determined with special polarographic microelectrodes inserted into the eye. P O 2 profiles established in vivo by driving electrodes through the entire retina yielded average P O 2 values between 10 mmHg (1.3 kPa) at the inner retinal surface and 382 mmHg (50.9 kPa) close to the outer retinal limit (Bruch's membrane). According to estimates on the basis of the diffusion distances determined from sections of the retina(∼436 μm at the site of P O 2 measurement)and literature data on specific oxygen consumption, the in vivo determined values would be sufficient to cover the oxygen demand of the retina with some safety margin. For a clear and direct in-tissue-test as to the involvement of the Root effect, an isolated in vitro eye preparation was established in order to avoid the problem of indirect blood supply to the eye from the dorsal aorta only via the pseudobranch, a hemibranch thought to modulate blood composition before entry of the eye. Any humoral effects (e.g. catecholamines)were eliminated by perfusing isolated eyes successively with standardized red blood cell (RBC) suspensions in Ringer, using trout (with Root) and human(lacking any Root effect) RBC suspension. To optimize perfusate conditions for maximal Root effect, the Root effect of trout RBCs was determined in vitro via graded acidification of individual samples equilibrated with standardized gas mixtures. During perfusion with trout RBC, P O 2 at the outer retinal limit was 99 mmHg(13.2 kPa), but fell by a factor of 3.3 upon perfusion with human RBC in spite of higher total oxygen content ( T O 2 2.8 for trout vs 3.9 mmol l -1 for human RBC). Upon reperfusion with trout RBC, P O 2 was restored immediately to the original value. This regularly observed pattern indicated a highly significant difference ( P =0.003) between perfusion with trout (with Root effect;high retinal P O 2 ) and perfusion with human (no Root effect; low retinal P O 2 ) RBC suspension,thus clearly demonstrating that the Root effect is directly involved and a crucial prerequisite for the enhancement of P O 2 in the retina of the teleost eye.

Changes in the rates of oxygen consumption and ammonium excretion, in intra- and extracellular acid-base status and in the rate of H+-equivalent ion transfer between animals and ambient water were measured during environmental hypercapnia in the peanut worm Sipunculus nudus. During exposure to 1 % CO2 in air, intracellular and coelomic plasma PCO2 values rose to levels above those expected from the increase in ambient CO2 tension. Simultaneously, coelomic plasma PO2 was reduced below control values. The rise in PCO2 also induced a fall in intra- and extracellular pH, but intracellular pH was rapidly and completely restored. This was achieved during the early period of hypercapnia at the expense of a non-respiratory increase in the extracellular acidosis. The pH of the extracellular space was only partially compensated (by 37 %) during long-term hypercapnia. The net release of basic equivalents under control conditions turned to a net release of protons to the ambient water before a net, albeit reduced, rate of base release was re-established after a new steady state had been achieved with respect to acid-base parameters. Hypercapnia also affected the mode and rate of metabolism. It caused the rate of oxygen consumption to fall, whereas the rate of ammonium excretion remained constant or even increased, reflecting a reduction of the O/N ratio in both cases. The transient intracellular acidosis preceded a depletion of the phosphagen phospho-l-arginine, an accumulation of free ADP and a decrease in the level of Gibbs free energy change of ATP hydrolysis, before replenishment of phosphagen and restoration of pHi and energy status occurred in parallel. In conclusion, long-term hypercapnia in vivo causes metabolic depression, a parallel shift in acid-base status and increased gas partial pressure gradients, which are related to a reduction in ventilatory activity. The steady-state rise in H+-equivalent ion transfer to the environment reflects an increased rate of production of protons by metabolism. This observation and the reduction of the O/N ratio suggest that a shift to protein/amino acid catabolism has taken place. Metabolic depression prevails, with completely compensated intracellular acidosis during long-term hypercapnia eliminating intracellular pH as a significant factor in the regulation of metabolic rate in vivo. Fluctuating levels of the phosphagen, of free ADP and in the ATP free energy change values independent of pH are interpreted as being correlated with oscillating ATP turnover rates during early hypercapnia and as reflecting a tight coupling of ATP turnover and energy status via the level of free ADP.

Blood distribution within the ventricle was analysed in acutely prepared turtles Trachemys scripta by measuring the oxygen concentration and flow rates of blood in the central vessels. Pulmonary (Qp) and systemic (Qs) blood flow rates were similar when total cardiac output (Qtot) was below 40 ml min-1 kg-1. Above this value, increments of Qtot were directed to the pulmonary circuit, with Qs levelling off at approximately 20 ml min-1 kg-1. When Qtot was larger than 40 ml min-1 kg-1, the systemic circuit was almost exclusively perfused by left atrial blood and systemic venous return was almost all directed towards the lungs. Blood oxygen levels and flow rates were consistently higher in the right aorta than in the left aorta. Blood movement within the ventricle, coupled with differences in ejection timing, is probably the decisive factor determining this pattern of blood distribution in the turtle heart.

The mechanism of cardiac shunting in reptiles is controversial. Recent evidence suggests that a right-to-left shunt in turtles results primarily from a washout mechanism. The mechanism that accounts for left-to-right (L-R) shunting is unresolved. This study used haemodynamic analysis and digital subtraction angiography to determine the mechanism of L-R cardiac shunting in the turtle Trachemys (Pseudemys) scripta. Animals were instrumented with ultrasonic blood flow probes (Transonic Systems, Inc.) for the measurement of total pulmonary blood flow and total systemic blood flow. In addition, catheters were inserted into the common pulmonary artery (PA), the systemic arteries, the left atrium and right atrium. These catheters were used for the measurement of blood pressure or for the infusion of radio-opaque material. Haemodynamic conditions were altered by electrical stimulation of the afferent (VAF) or efferent vagal nerves or by infusion of vasoactive drugs. Under control conditions, the peak systolic pressure in the systemic arteries was slightly higher than that in the PA (30.6 versus 28.3 mmHg; 4.08 versus 3.77 kPa), whereas diastolic pressure in the PA was significantly less than that in the systemic arteries (9.8 versus 24.4 mmHg; 1.31 versus 3.25 kPa). During VAF stimulation, the peak systolic pressures in the PA and aortae almost doubled. Diastolic pressure in the systemic arteries also doubled, but it increased by only 45% in the PA. Ejection of blood into the PA preceded that into the left aorta by 53 ms under control conditions. This difference increased (by as much as 200 ms) as the difference in the diastolic pressures between the two circulations increased during VAF stimulation. This resulted in the development of a large net L-R shunt. Under these conditions, digital subtraction angiography showed that the L-R shunt resulted from a combination of both washout and pressure mechanisms.

Previous studies of reptiles and amphibians have shown that changing the body temperature consistently produces transient changes in the respiratory exchange ratio (RE) and, hence, changes in whole-body CO2 stores, and that the extracellular fluid compartment contributes to the temperature-related changes in CO2 stores. The purpose of this study was to determine whether the intracellular fluid compartment contributes to the changes in CO2 stores in undisturbed resting cane toads. Increasing body temperature from 10 to 30 degrees C temporarily elevated RE, and returning body temperature to 10 degrees C temporarily lowered RE. The estimated average change in whole-body CO2 stores associated with the transient changes in RE was 1.0 +/- 0.8 mmol kg-1 (+/- S.D., N = 6). Plasma [CO2] and, thus, extracellular fluid [CO2], were unaffected by the temperature change. Plasma calcium levels were also unaffected, so that bone CO2 stores did not contribute to changes in whole-body CO2 stores. Intracellular [CO2] was determined for the lung, oesophagus, stomach, small intestine, liver, ventricle, red blood cells, skin and 14 skeletal muscles. [CO2] was significantly lower (P < 0.05) at higher temperature in 10 of these, and seven others, although not statistically significant (P > 0.05), had mean values at least 0.5 mmol kg-1 lower at the higher temperature. The average change in intracellular [CO2] for all tissues examined was -0.165 mmol kg-1 degrees C-1. We conclude that, in cane toads, the temperature-related transients in RE result from intracellular CO2 adjustments, that different tissues have unique intracellular CO2/temperature relationships, and that a combination of respiratory and ion-exchange mechanisms is used to adjust pH as temperature changes.

Proton-equivalent ion transfer processes between animals and ambient water were determined under normoxic control conditions during anaerobiosis and the subsequent recovery period in the marine worm Sipunculus nudus L. During anaerobiosis and recovery, transepithelial H + -equivalent ion transfer was generally correlated with changes in extracellular pH, with some disparities in ‘spring’ animals. The typical initial alkalosis induced by phosphagen cleavage during early anaerobiosis was reflected by a loss of basic equivalents. The acidosis, which developed later, reflecting production of acidic metabolic intermediates, resulted in a relatively small net extrusion of protons into the water. The coelomic acidosis during recovery was greatly exaggerated by the release of protons during phosphagen repletion and by the considerable elevation of P co 2 after normoxia had been reattained. The acidosis stimulated the net release of H + to the water at a rate several times higher than that during anaerobiosis. The efficient transfer of protons from the body fluids to the environmental water during recovery facilitated normalization of coelomic pH, long before protons dissociated from the large amounts of organic acids produced as anaerobic intermediates could be removed from the body fluids by metabolism. Although the transfer of net H + equivalents to the water coincided with coelomic acidosis, the rates of transfer during different periods of the experiment were primarily correlated with overall metabolic rate. Low net proton transfer rates associated with anaerobiosis were not sufficient to maintain acid-base parameters typical for normoxia, whereas re-establishment of aerobic conditions facilitated a greatly increased transepithelial H + transfer rate. These data suggest that the transfer capacity of the energy-consuming translocation mechanism may primarily be determined by the rate of metabolic turnover and, accordingly, by theamount of available energy.

The effects of changes in environmental temperature and oxygen tension on gill ventilation, arterial P O 2 , P CO 2 pH and [HCO 3 − ] were evaluated in carp ( Cyprinus carpio L.). Gill ventilation was measured continuously in specimens acclimated to 10 or 20°C, combining the method of electromagnetic flow determination with the application of a rubber mask technique. After establishing control values in airequilibrated water the environmental water P O 2 (Pw O 2 ) was reduced from about 150 mmHg (20 kPa) during control conditions to 110 or 75 mm Hg (14.7 or 10 kPa), respectively. Measurements of blood gases and acid-base parameters were performed repeatedly before, and 1 and 4 h after, initiation of hypoxia. Regardless of temperature, these moderately hypoxic conditions caused considerable and lasting increases in gill ventilation of about 70% (PW O 2 =110 mm Hg/l4.7kPa) or 180% (Pw O 2 =75 mm Hg/10kPa), relative to the respective normoxic control values of about 50 ml kg −1 min −1 at 10°C and 230 ml kg −1 min −1 at 20°C. These increases in ventilation reduced P CO 2 substantially, resulting in a rise in pHa by about 0.1 units at Pw O 2 of 110 mmHg (14.7 kPa) and by about 0.2 units at Pw CO 2 of 75 mmHg (lOkPa). Arterial P O 2 was low under normoxic conditions at both temperatures (≈15 mmHg, ≈2kPa). During hypoxia, Pa O 2 was marginally reduced, whereas the arterial O 2 content and saturation remained at normoxic levels, mainly because of the increase in the blood O 2 -affinity induced by respiratory alkalosis. Thislack of any clear relationship between arterial O2 content and ventilatory response to moderate hypoxia contrasts with previously reported data for trout, and supports the hypothesis that a change in P O 2 is an adequate stimulus for the adjustment of ventilation in carp.

The patterns of blood flow through the complex circulation of the tiger salamander, Ambystoma tigrinum, were investigated during aquatic normoxia and hypoxia by application of the microsphere technique. The distribution of differently labelled microspheres injected into the bloodstream towards the left and right atria, respectively, was used to evaluate the role of the ductus arteriosus in lung perfusion before, during and after metamorphosis, as well as the general contribution of right and left atrial outputs to the blood flow in gills and lungs in neotenic and postmetamorphic animals. The distribution patterns of radioactive microspheres among pulmonary, branchial and systemic tissues indicated that the ductus arteriosus is the major pulmonary perfusion pathway in neotenic and metamorphosing animals, whereas after metamorphosis the main perfusion pathway is down the entire length of the pulmonary artery. In neotenes, the ductus arteriosus becomes even more important during aquatic hypoxia. The anterior branchial arches receive blood richer in pulmonary venous blood than the posterior arches. Approximately 26% of left atrial output and 36% of right atrial output perfuses the brachial respiratory lamellae during normoxia in neotenes. Severe aquatic hypoxia appears to increase the fraction of cardiac output flowing to the lung and decrease the fraction flowing into the first branchial arch in neotenes. This decrease into the first arch may facilitate lung perfusion and also reduce branchial O2 loss. In postmetamorphic animals, approximately 55% of right atrial output and 32% of left atrial output is directed to the lungs. The flow patterns in postmetamorphic animals remain unaffected by aquatic hypoxia.

Freshwater-acclimated rainbow trout were chronically and non-occlusively cannulated in the dorsal aorta (DA), sinus venosus (SV) and branchial vein (BV), the latter returning the blood perfusing the central venous sinus (CVS) of the gill after being shunted away from the systemic circuit. After recovery, blood samples from these three sites were analysed for haematocrit (Hct) and [Hb]. Branchial venous blood was found to have considerably lower Hct and [Hb] (Hct = 3.5 +/− 3.1%; [Hb] = 1.04 +/− 0.75 g 100 ml-1) than systemic blood (DA: Hct = 24.3 +/− 8.9%, [Hb] = 6.54 +/− 2.90 g 100 ml-1; SV: Hct = 23.1 +/− 8.8%, [Hb] = 6.15 +/− 2.55 g 100 ml-1; means +/− S.D. N = 8), which strongly suggests that plasma skimming occurred at arteriovenous anastomoses and arterioles draining into the CVS. The partitioning of cardiac output, calculated on the basis of the [Hb] data, showed that the systemic flow accounted for 93 +/− 4.6% (N = 7) of the total cardiac output with only 7 +/− 4.7% of cardiac output being diverted into the CVS, thus bypassing the systemic vasculature. Previous results using in vitro experiments are compared with the present data in an evaluation of the usefulness of the isolated perfused gill and head preparations in the experimental analysis of circulatory physiology in fish gills.

Specimens of Bufo marinus were exposed to aerial and aquatic hypercapnia (5% CO2) in a closed, water recirculation system to evaluate mechanisms involved in the compensation of a respiratory acidosis in these animals. Arterial PCO2 was elevated from about 9 mmHg (1 mmHg = 133.3 Pa) to 35 (1 h) and 37 mmHg (2 h), and gradually approached about 40 mmHg (24 h of hypercapnia). The typical hypercapnia-induced reduction in plasma pH from about 7.9 to below 7.4 was partially offset, at least during the first hours of hypercapnia, by a reduction in the inspired/arterial PCO2 difference, presumably brought about by pulmonary hyperventilation. The predominant contributor to extracellular pH compensation, however, was a net gain of bicarbonate from the environment, mainly facilitated by ammonia excretion. Bicarbonate originating from the environment was accumulated in the body fluids, increasing the plasma concentration from the control of about 9 to 36 mmol l-1 after 24 h. Extracellular pH was compensated to only about 30% of the shift expected at constant bicarbonate level and, according to the steady reduction of pH, non-bicarbonate buffering of CO2 also contributed significantly to the elevation of bicarbonate. This relatively poor pH compensation (compared with fishes) could not be improved either by direct administration of bicarbonate into the bloodstream or by increased environmental ion concentrations. It is concluded that the availability of bicarbonate is not a limiting factor for pH compensation during hypercapnia, and that the inability of Bufo to accumulate bicarbonate to concentrations sufficient for better hypercapnia compensation is based on a constitutional ‘bicarbonate threshold’ of the resorbing and retaining structures for acid-base-relevant ions.

Blood gases, and parameters of the extracellular and intracellular acid-base status, were measured in the anuran amphibians Bufo marinus and Xenopus laevis acclimated to temperatures of 10, 20 and 30°C for 12 days. Arterial P O O2 rose with temperature so that approximately constant oxygen saturation of the blood was maintained, a phenomenon explained on the basis of models for O 2 transport in animals with central vascular shunts and temperature-dependent shifts in O 2 equilibrium characteristics. Arterial plasma pH of both species varied inversely with temperature, the pH/temperature coefficient being not significantly different from that required for constant relative alkalinity or dissociation of imidazole. The change in plasma pH was brought about mainly by changes in P CO CO2 although plasma bicarbonate concentration also changed significantly. Intracellular pH/temperature relationships were found to be non-linear in most of the tissues. There was considerable variability among body tissue compartments and between the two species. These data confirm that the various tissue compartments in ectotherms maintain unique ΔpH/Δt relationships, and indicate that measurement of extracellular pH as a function of temperature is not a good indicator for alphastat-type, temperature-dependent, acid-base regulation.

The distribution of pulmocutaneous heart output to lungs and skin was determined in non-anaesthetized, fully recovered bullfrogs (Rana catesbeiana) by application of the microsphere method in order to study the modulation of blood flow to different gas exchange sites in amphibians during environmental air and water hypoxia. The relative perfusion of various skin areas was found to be rather heterogeneously distributed with an over-proportionately high blood flow to the ventral body surface. This distribution of flow among different skin areas remained unaffected by any type of environmental hypoxia. The relative perfusion of lungs and skin, however, was significantly affected by the pattern of environmental oxygen partial pressure. The relative lung perfusion (approximately equal to 80% of pulmocutaneous flow in normoxic control conditions) was increased during water hypoxia, and reduced with lowered inspired PO2. This mechanism could be interpreted as a readjustment of blood flow towards the gas exchange site with higher oxygen partial pressure, but may also represent a mechanism to prevent oxygen loss from the body stores at gas exchange sites of low oxygen tension.

To study both temporal and quantitative effects of hypercapnia on the extent of pH compensation in the arterial blood, specimens of carp (Cyprinus carpio) were exposed to a PCO2 of about 7.5 mmHg (1 mmHg = 133.3 Pa) (1% CO2) in the environmental water for several weeks, and a second group of animals was subjected to an environmental PCO2 of about 37 mmHg (5% CO2) for up to 96 h. A third series of experiments was designed to test the possibility that infusion of bicarbonate would increase the extent of plasma pH compensation. Dorsal aortic plasma pH, PCO2 and [HCO3-], as well as net transfer of HCO3- -equivalent ions, NH4+, Cl- and Na+, between fish and ambient water, were monitored throughout the experiments. Exposure to environmental PCO2 of 7.5 mmHg resulted in the expected respiratory acidosis with the associated drop in plasma pH, and subsequent compensatory plasma [HCO3-] increase. The compensatory increase of plasma bicarbonate during long-term hypercapnia continued during 19 days of exposure with plasma bicarbonate finally elevated from 13.0 mmoll-1 during control conditions to 25.9 mmoll-1 in hypercapnia, an increase equivalent to 80% plasma pH compensation. Exposure to 5% hypercapnia elicited much larger acid-base effects, which were compensated to a much lesser extent. Plasma pH recovered to only about 45% of the pH depression expected at constant bicarbonate concentration. At the end of the 96-h exposure period, plasma [HCO3-] was elevated by a factor of 2.5 to about 28.2 mmoll-1. The observed increase in plasma bicarbonate concentration during 5% hypercapnic exposure was attributable to net gain of bicarbonate equivalent ions from (or release of H+-equivalent ions to) the environmental water. Quantitatively, the gain of 15.6 mmol kg-1 was considerably larger than the amount required for compensation of the extracellular space, suggesting that acid-base relevant ions were transferred for compensation of the intracellular body compartments. The uptake of bicarbonate-equivalent ions from the water was accompanied by a net release of Cl-and, to a smaller extent, by a net uptake of Na+, suggesting a 75% contribution of the Cl-/HCO-3 exchange mechanism. Infusion of bicarbonate after 48 h of exposure to 7.5 mmHg PCo2 had only a transient effect on further pH compensation. The infused bicarbonate was lost to the ambient water, and pre-infusion levels of bicarbonate were reattained within 24 h. Repetition of the infusion did not result in a notable improvement of the acid-base status.(ABSTRACT TRUNCATED AT 400 WORDS)

The effects of hypercapnia on the arterial acid-base status of the Tegu lizard, Tupinambis nigropunctatus (Spix), were studied at 25°C. Arterial P CO CO2 increased over the first 2 h of hypercapnia causing a fall in arterial plasma pH (pH a ) of about 0.17 units with ≈4% CO 2 and about 0.3 units with ≈7% CO 2 . In both conditions, plasma pH increased slightly (≈0.02units) between 2 and 72 h. Plasma [HCO 3 − ] rose during the initial increase of Pa CO CO2 (by approx. 5.5 mmol with ≈7% CO 2 ; approx. 1.9 mmoll −1 with ≈4% CO 2 ) during the first 2h, and further increased by 4 mmoll −1 between 2 and 72 h of hypercapnia, while Pa CO CO2 did not change. The increases of plasma [HCO 3 − ] resulted in a recovery of pH, by 38 or 32 % (≈4 and ≈7% CO 2 , respectively) relative to the fall of pH, that would occur at constant [HCO 3 − ]. The limited and incomplete compensation of pH a during environmental hypercapnia is consistent with data for other air-breathing ectothermic vertebrates, and contrasts with the typical response of water-breathing fish, in which compensation is usually complete. Note:

Freshwater turtles ( Chrysemys picta bellii Gray) were acclimated to temperatures of 5, 10, 20 and 30°C for at least 12 days, and pulmonary ventilation, oxygen uptake and arterial pH, P CO CO2 and P O O2 were determined in completely unrestrained specimens. Oxygen uptake (V · O O2 ) increased overproportionately (6.7-fold) as compared to pulmonary ventilation (V · 1 , 4.4-fold) when the temperature increased from 10 to 30°C. The observed rise in arterial P CO CO2 from 13 (5°C) to 32mmHg (30°C) was the result of a decrease in V · 1 /V · O O2 , whereas an increase of arterial P O O2 from 12Torr at 5°C to about 60Torr at 20 and 30°C mainly resulted from the effects of intracardiac blood shunting combined with temperature-dependent shifts of the oxygen dissociation curve. Arterial pH fell with rising temperature significantly less (ΔpH/Δt =−0.010U/°C) than required for constant relative alkalinity and for constant dissociation of imidazole. The changes of cerebrospinal fluid pH with temperature, calculated from the mean arterial P CO CO2 values, were even smaller [ΔpH/Δt CSF = −0.008). It is concluded that the observed temperature dependence of the acid-base status is not in agreement with the alphastat hypothesis.

Specimens of Conger conger (L.) were exposed to environmental hypercapnia in a closed recirculating seawater system. Arterial plasma pH, PCO2 and bicarbonate concentration, as well as the net transfer of bicarbonate and ammonia between fish and ambient seawater, were monitored for 30 h of hypercapnia. The initial hypercapnia-induced reduction of arterial pH by about 0.4 pH units was restored to near control values within 10 h of hypercapnia by compensatory elevation of plasma bicarbonate concentration. The continuous rise in extracellular bicarbonate from about 5 to 22 mM during this time was the result of two different mechanisms. Initially, there was a net bicarbonate transfer from the intracellular space to the extracellular compartment until the net uptake of bicarbonate from the seawater started. The amount of bicarbonate originally transferred to the extracellular space was then returned to the intracellular compartment and finally the changes in both extracellular and intracellular pH were compensated by bicarbonate taken up from the environmental seawater. Since the ammonia excretion was not increased during hypercapnia and the pattern of plasma electrolyte concentrations does not favour the H+/Na+ ion exchange mechanism, it is concluded that the additional bicarbonate is gained by active HCO3-/Cl- ion exchange against the electrochemical gradient between fish and seawater.

Intracardiac shunting was studied in unanaesthetized and unrestrained specimens of Varanus exanthematicus by simultaneous injection of radioactively labelled microspheres (15 micron) into the right and left atria. Lung ventilation was monitored by intratracheal pneumotachography. It was found that intracardiac shunting was not significantly affected by the spontaneously occurring periods of ventilation and apnoea: the right-to-left shunt averaged 29 and 31%, respectively, and the left-to-right shunt was 11% in both conditions. The observed shunting, although rather constant with time and independent of the ventilatory state, varied in different individuals. Anatomical studies and intracardiac pressure measurements revealed that, in spite of crocodilian-like systolic pressure separation between pulmonary and systemic circulation (based on the muscular ridge, ‘Muskelleiste’, between cavum venosum and cavum pulmonale), the cavum venosum is shared by both the pulmonary and the systemic circulation. Intracardiac shunting appears to be mainly due to wash-out of the cavum venosum: blood remaining in this chamber at the end of systole (oxygenated) or at the end of diastole (deoxygenated) is washed into the respective ‘inadequate’ vascular bed during the next half-cycle of heart action. Thus the extent of intracardiac shunting is expected to depend primarily on the volume and the changes in volume of the cavum venosum during the cardiac cycle.

Specimens of the larger spotted dogfish (Scyliorhinus stellaris) were electrically stimulated to exhaustion in a closed seawater recirculation system. The production of large quantities of lactic acid by anaerobic metabolism and the resultant efflux of the dissociation products, H+ and lactate, from the white musculature resulted in severe acid-base disturbances and in increases in plasma lactate concentration, the two effects having extremely different time courses. Plasma pH and bicarbonate were maximally depressed 15–30 min after exercise, whereas peak lactate concentrations of up to 30 mM were not attained before 4–8 h after exercise. The acid-base status were restored to normal 10–14 h after exercise, long before the aerobic processing of surplus lactic acid was complete 22–30 h after exercise. This behaviour can be explained on the basis of an interaction of transfer rates, buffer values and equilibria between intracellular and extracellular compartments with the transient net transfer of surplus H+ ions to the environmental water. About half of the original quantity of H+ was transferred net to the environment via the branchial epithelium during the first 8–10 h, and it was later taken up again at the rate of aerobic lactic acid processing in the metabolism of the fish, whereas a transfer of lactate was not observed at any time during the experiment. As a result, the distribution patterns of H+ and lactate differed from each other and varied with time elapsed after anaerobic exercise, leading to the apparent ‘H+ ion deficit’ which has been observed in the blood of several fish species during lactacidosis. Net transfer of H+ ions to the environment facilitates rapid normalization of the acid-base status long before the original stress, lactic acid, is removed from the organism and thus represents an effective regulatory mechanism for the defence of the internal milieu in fish.

In the tropical fresh water fish, Synbranchus marmoratus, transition from water breathing to air breathing, induced by reduction of oxygen partial pressure (PO2) in the environmental water below 16 mmHg, causes a considerable rise in the arterial partial pressure of carbon dioxide (PCO2), from 5.6 to 26 mmHg on the average (half time of the rise between 2 and 6.5 h). The associated fall in arterial plasma pH by about 0.6 units is not compensated by an increase in plasma bicarbonate concentration, whereas the intracellular pH of white skeletal muscle and heart muscle is kept almost constant by elevation of the intracellular bicarbonate concentration. The additional bicarbonate is generated by intracellular non-bicarbonate buffering, and by net transfer into the intracellular space of bicarbonate formed by buffering in blood. Only a relatively small quantity of bicarbonate is taken up from environmental water. This type of acid-base regulation, with almost complete intracellular pH compensation and only minor bicarbonate uptake (equivalent H+ release or OH- uptake) from water, is attributed to several factors. Probably the most important of these is the lack of continuous contact of the gills, which are the main site of ion transfer processes, with the environmental water during air breathing. Regardless of the mechanisms involved, this particular strategy of acid-base regulation provides a constant milieu for the intracellular structures and demonstrates the prevalence of intracellular over extracellular acid-base regulation.