Effects of anoxia on the activities of pyruvate kinase and phosphoenolpyruvate carboxykinase,and the production of lactate and succinate in the intertidal pulmonate Onchidium tumidium

Onchidium tumidium, an intertidal pulmonate, has evolved to depend mainly on the formation of succinate, rather than lactate and opines, to survive in anoxia. For our study O. tumidum were collected from the mud flats of the mangrove swamp at Mandai, Singapore between 1988 and 1991. After 24 h of anoxic exposure, the lactate and succinate contents of the anoxic individuals were approximately 10 and 150 times, respectively, the corresponding values of the normoxic individuals. Alanine and acetate accumulations also occurred during anoxia, though to a much lesser extent. No propionate or octopine was detected. The depletion in aspartate content in O. tumidium could not account for the amount of succinate accumulated during anoxia. The succinate formed might have originated from glycogen involving the flow of carbon through the phosphoenolpyruvate (PEP) branch point of glycolysis. In support of such a hypothesis, results indicate that there was a decrease in the affinity of pyruvate kinase from O. tumidium exposed to 24h of anoxia to PEP to facilitate succinate formation through phosphoenolpyruvate carboxykinase (PEPCK). In comparison, the affinity of PEPCK from O. tumidium exposed to anoxia to PEP was apparently unaltered.     

Regulation of glycolytic enzymes in the marine invertebrateHalicryptus spinulosus (Priapulida) during environmental anoxia and exposure to hydrogen sulfide

A particularly strong reduction of metabolic activity is a precondition for long-term survival ofHalicryptus spinulosus von Siebold under anoxic habitat conditions because of its relatively low fuel reserves (mainly glycogen). The present study analyses the mechanism of this metabolic slow-down. For this purpose the effects of environmental anoxia and exposure to hydrogen sulfide on the activity and selected kinetic properties of glycolytic enzymes [glycogen phosphorylase (GP), pyruvate kinase (PK)] and the concentrations of fructose-2,6-bisphosphate in the body wall ofH. spinulosus were analysed. Anoxia and hydrogen sulfide exposure stimulated modifications of the properties of the enzymes, in both cases due to probable covalent modification of the enzyme proteins. Under both conditions phosphorylase activity was depressed by about 1/3, the result of changes in the percentage of enzyme in the activea-form as well as the total amount of enzyme activity expressed (a +b). Effects of anoxia on the properties of pyruvate kinase included reducedV _max , decreasedS _0.5 for phospho-enolpyruvate, changes inK _a for fructose-1,6-bisphosphate (an initial decrease was followed by a later increase). TheI ₅₀ forL-alanine of PK was extremely reduced under anoxia and showed an even greater sensitivity to the presence of hydrogen sulfide. Anoxia stimulated a slight reduction in the content of fructose-2,6-bisphosphate, whereas exposure to hydrogen sulfide caused a dramatic decrease of this allosteric activator of phos-phofructokinase. The study gives evidence that mechanisms of glycolytic rate depression are conserved within a wide variety of vertebrate and invertebrate phyla. With two exceptions (fructose-2,6-bisphosphate levels and alanine inhibition of PK) the responses to hydrogen sulfide were the same as those to anoxia, suggesting that at a metabolic level, the consequences of each stress on energy metabolism are similar.     

Biochemical adaptations of the mudskipper Boleophthalmus boddaerti to a lack of oxygen

Specimens of the mudskipper Boleophthalmus boddaerti, collected along the estuarine canal at Pasir Ris, Singapore between April 1987 and December 1989, were examined in the laboratory. After being exposed to environmental hypoxia for 6 h, no accumulation of lactate, alanine or succinate was observed in the muscle tissue of B. boddaerti. In addition, the blood lactate content and muscle creatine phosphate (CrP), adenosine triphosphate (ATP) and glycogen contents were not significantly different from those of the control. During normoxic recovery, oxygen debt repayment was only a small fraction of the oxygen deficit incurred during the 6 h of hypoxic exposure. Therefore, it would appear that B. boddaerti coped with environmental hypoxia by undergoing metabolic rate reduction. After anoxic exposure, the CrP, ATP and adenosine diphosphate (ADP) contents in the muscle tissue of B. boddaerti were significantly lower than the corresponding control values. Lactate accumulated in the blood and muscle tissue of the anoxic fish though the muscle energy charge and glycogen content remained constant. Hence, B. boddaerti was able to cope with such a period of anoxia without increasing its glycolytic rate. Succinate, which was not detected in the muscle tissue of the control and hypoxia-exposed fish, accumulated in significant quantities in the muscle tissue of B. boddaerti exposed to environmental anoxia and functional hypoxia. In the case of functional hypoxia, there was a significant decrease in the muscle glycogen content, and the muscle lactate content increased 17.3-fold, indicating that glycolysis was activated to provide the source of energy during muscular exercise.     

Effect of fructose 2,6-bisphosphate and ammonium ions on AMP-mediated activation of phosphofructokinase from the posterior adductor muscle of the sea musselMytilus galloprovincialis

The effects of different modulators on the phosphofructokinase (PFK) activity from the posterior adductor muscle of the sea musselMytilus galloprovincialis Lmk. were studied in mussels collected from N.W. Spain in spring/summer, 1988. Adenosine monophosphate (AMP), fructose 2,6-bisphosphate (Fru 2,6-P₂) and ammonium ions individually activated PFK. AMP and Fru 2,6-P₂ reduced ATP-mediated inhibition and the value ofS _0.5 [concentration of fructose 6-phosphate (Fru 6-P) for half-maximum velocity], whereas ammonium ions increasedV _max (the rate at the saturating concentrations of Fru 6-P). Fru 2,6-P₂ and ammonium ions both enhanced the effect of AMP, but in different ways: Fru 2,6-P₂ reduced apparentK _m for AMP (concentration of AMP for half-maximum activation) without affectingK _m , whereas ammonium ions only altered the latter. The sharp increases in the values ofS _0.5 for Fru 6-P and of apparentK _m for AMP that are caused by a drop in pH were reduced by the presence of Fru 2,6-P₂, but not by ammonium ions. The results shows that Fru 2,6-P₂ may contribute to maintain the PFK activity during hypoxia periods, whereas ammonium ions (in the presence of high AMP concentrations) may activate the enzyme during the first few hours of post-hypoxia re-immersion.     

Ammonia production and kinetic properties of glutamate dehydrogenase in the sipinculid Phascolosoma arcuatum exposed to anoxia

The amounts of total NH ₄ ⁺ detected in the external media in which Phascolosoma arcuatum had been exposed to various periods of anoxia were significantly greater than those in which the worms were exposed to normoxia for a similar period. The increased NH ₄ ⁺ production by P. arcuatum during anoxic exposure was unlikely to be due to an increased catabolism of adenine nucleotides or urea. In contrast, there were significant decreases in the concentrations of several free amino acids in the coelomic plasma and body tissues of individuals during the 48 h of anoxic exposure. The amount of NH ₄ ⁺ produced by the anoxic P. arcuatum could be accounted for by the decreases in the concentrations of aspartate or glycine. Increases in the catabolism of free amino acids (FAA), leading to the increased production of NH ₄ ⁺ , in P. arcuatum during anoxia were supported by the detection of significant changes in the kinetic properties of glutamate dehydrogenase (GDH), in the deaminating direction, from worms exposed to anoxia for 48 h. The apparent increase in the affinity of GDH from the anoxic worm to glutamate would bring about a greater deaminating activity at physiological concentrations of ths substrate. P. arcuatum used in these experiments were collected from the mangrove swamp at Mandai, Singapore between 1990 and 1993.     

Energy metabolism in the foot of the marine gastropod Nassa mutabilis during environmental and functional anaerobiosis

This study concerns the effects of oxygen deprivation due to incubation in oxygen free sea water (environmental anoxia) or exercise (functional anoxia) and of exposure to air on the mode of energy production in the foot of the whelk Nassa mutabilis. Additionally, energy metabolism of the foot muscle was investigated during exercise after different anoxia periods and during the subsequent recovery period. During environmental anoxia, phosphoarginine, glycogen and aspartate are broken down as substrates and alanine and succinate are formed as products. There was no production of D-lactate or octopine. The energy charge value fell after 24-h anoxia. Exposure to air resulted in only small changes in phosphoarginine and alanine levels, suggesting that oxygen uptake was impaired in the first phase of air exposure but that, later, aerial respiration kept pace with the energy demand. Exercise caused a dramatic decrease of phosphoarginine concentration, coupled with glycolytic ATP production via octopine formation. In the recovery period (after exercise), the level of phosphoarginine was rapidly restored. An anaerobic component was evident during recovery as shown by the accumulation of D-lactate. Thus, both terminal dehydrogenases, octopine- and lactate dehydrogenase, are active in the muscle, but under different physiological conditions. Octopine formation also took place when the whelks were subjected to exercise after 4 or 24 h of anoxia. In this case, glycolysis provided between 70 and 90% of the energy required since the phosphagen store had already been depleted during the anoxic period. When the work load was increased (greater number of leaps), it became evident that the action of arginine kinase and octopine dehydrogenase are not closely linked. First there was an increase of arginine and then later a condensation of arginine with pyruvate to form octopine.     

Cyclic AMP-dependent protein kinase: role in anoxia and freezing tolerance of the marine periwinkle Littorina littorea

A key regulatory mechanism underlying the switch between aerobic and anaerobic metabolism amongst anoxia-tolerant marine molluscs is reversible protein phosphorylation. To assess the role of cAMP-dependent protein kinase (PKA) in aerobic–anaerobic transitions, the effects of anoxia on the activity and subcellular distribution of PKA were assessed in foot and hepatopancreas of the marine periwinkle, Littorina littorea. Exposure to N₂ gas at 5 °C caused a rapid decline in the percentage of total enzyme present as the free catalytic subunit (PKAc) in both tissues; the percentage of PKAc fell from ∼30% in controls to 3% after 1 h anoxia and remained low over 72 h. Total PKA also fell by 30% after 72 h anoxia in hepatopancreas but rebounded during aerobic recovery. Freezing at −8 °C elicited parallel results for both percentage of PKAc and total PKA, suggesting that PKA responses to freezing were stimulated by the ischemia that develops when hemolymph freezes. Anoxia also led to a shift in PKA subcellular distribution in hepatopancreas (but not in foot), the percentage of total PKA activity associated with the nuclear fraction dropping from 25% in controls to 8% in 12 h anoxic snails with opposite changes in the cytosolic fraction. The catalytic subunit (PKAc) of foot PKA was purified to a final specific activity of 63.5 nmol phosphate transferred per minute per milligram protein. Enzyme properties included a molecular weight of 33 to 35 kDa, an activation energy from Arrhenius plots of 65.1 ± 4.8 kJ mol⁻¹, and substrate affinity constants of 151 ± 6 μM for the phosphate acceptor, Kemptide, and 72 ± 9 μM for Mg.ATP. Activity was strongly reduced by mammalian PKA inhibitors (H-89, PKA-I), by neutral chloride salts (I₅₀ values 165 to 210 mM) and by NaF (I₅₀ 62 mM). Reduced PKA activity under anoxic or freezing conditions would facilitate the observed suppression of the activities of numerous enzymes that are typically PKA-activated and thereby contribute to the overall anoxia-induced metabolic rate depression. Received: 19 November 1997 / Accepted: 30 September 1998     

Lactate formation in Callianassa californiensis and Upogebia pugettensis (Crustacea: Thalassinidea)

The ability to accumulate lactate as a result of laboratory exposure to anoxia was examined in 2 species of mud-dwelling shrimp, the ghost shrimp Callianassa californiensis and the mud shrimp Upogebia pugettensis. Hemolymph lactate accumulated to a much greater degree in the mud shrimp, even though the overall levels of lactate dehydrogenase activity were similar in tissues of the two species. Carbohydrate reserves, estimated as glycogen, were not significantly depleted in muscle or midgut gland of either species as a result of anoxic stress. The results are discussed in relation to possible metabolic strategies employed to cope with the hypoxic environment of these crustaceans.     

Metabolic responses of the prawns Palaemon elegans and P. serratus (Crustacea: Decapoda) to acute hypoxia and anoxia

Using prawns, Palaemon elegans (Rathke) from intertidal pools on the Isle of Cumbrae, Scotland, and P. serratus (Pennant) from the subtidal at Plymouth, England, some metabolic responses to hypoxia and anoxia have been studied. P. elegans was found to have a greater tolerance of severe hypoxia than P. serratus. Tolerance of totally anoxic conditions, however, was limited to only 4 h in P. elegans and to approximately 1 h in P. serratus. exposure to moderate hypoxia (30 torr) resulted in little change in the concentration of L-lactate in the blood or in the tissues of either P. elegans or P. serratus. When exposed to extreme hypoxia (10 or 5 torr for P. elegans), however, there was a progressive increase in the concentration of L-lactate in the blood and in the tissues of both species. After normoxic conditions had been restored, the concentration of L-lactate in the blood and in the tissues returned to normal resting levels more rapidly in P. elegans than in P. serratus. Under hypoxic conditions, both P. elegans and P. serratus showed an increase in the concentration of blood glucose and a slight reduction in the glycogen content of the tissues. The concentrations of blood glucose and of tissue glycogen returned to normal levels within 6 h of the prawns being returned to normoxic conditions. The results of an in situ study in April and August 1986 to examine the metabolic responses of P. elegans to the hypoxic conditions normally experienced in high-shore rock pools are also presented. The ecological significance of the differing abilities of these species to survive hypoxic exposure is discussed.     

Effects of arsenic on survival and metabolism of Crangon crangon

Metabolic changes and toxic effects in Crangon crangon (collected during summer 1989 near Helsingør, Denmark and at Kulhuse in the Isefjord) were observed during exposure to different arsenate concentrations. Survival, respiration, and concentrations of hemolymph glucose, muscle glycogen andadenine nucleotides were measured. C. crangon was very tolerant of arsenate; survival was only affected at arsenate concentrations >25 ppm. Small individuals (0.2 to 0.29 g wet weight) were less tolerant than medium-sized and large individuals (0.5 to 0.59 g and 0.8 to 0.89 g wet weight), e.g. at 50 ppm the LT₅₀ for small individuals was 180 h compared to 343 h and 360 h in middle-sized and large individuals. The relation between size and toxicity is probably due to the change in surface/volume ratio. Under clean conditions the respiration of C. crangon showed a clear relation between routine MO₂ (O₂ in mlg^-1 h^-1) and wet weight (r²=0.89; P<0.001). During shortterm exposure to arsenate (10 and 50 ppm), the respiratory rates (MO₂) seemed to decrease in a size-dependent manner. The concentrations of blood glucose and muscle glycogen did not change in response to arsenate exposure. The concentrations of adenylate phosphates were high and constant throughout the experiments. C. crangon had maximal adenylate energy charge (AEC) values of ca. 0.8. No decrease in AEC due to the exposure to arsenate was observed.     

Carbohydrate metabolism during anoxia and post-anoxia recovery in Chasmagnathus granulata crabs maintained on high-protein or carbohydrate-rich diets

We assessed the effects of anoxia exposure and recovery on glycogen synthesis and mobilization, glucose uptake, and on the enzymes that control carbohydrate metabolism in the hepatopancreas of Chasmagnathus granulata crabs receiving either a carbohydrate-rich (HC) or a high-protein diet (HP). In both dietary groups, anoxia led to a reduction in glucose uptake and in glycogen synthesis, and to an increase in hepatopancreas glycogen mobilization and in hemolymph glucose concentration. During the first 4 h of exposure to anoxia, total glycogen phosphorylase (GPT) and a form activity increased in HP and HC crabs, leading to a decrease in hepatopancreas glycogen concentration. During recovery, HP and HC crabs rapidly restored the hemolymph glucose levels to pre-anoxia concentrations. In HC crabs, incorporation of ¹⁴C from glucose into glycogen increased gradually after 12 h in normoxia, leading to restoration of glycogen concentration. Also during recovery, the ratio of glycogen synthase I (GSI) to glycogen phosphorylase a (GPa) increased in the HC group. In turn, recovering HP crabs had two peaks of glycogen synthesis, related with two peaks in the ratio of GSI to GPa. Consequently, no mobilization of ¹⁴C-glycogen occurred in recovering HP animals. Anoxia in C. granulata induces a marked decrease in the synthesis of carbohydrate reserves that is accompanied by an increase in glycogen mobilization and in circulating glucose levels. During the recovery period, there is an activation of endergonic processes which cause a decrease in hemolymph glucose levels. In C. granulata, glycogen metabolism seems to be controlled by the ratio of the GSI form to the GPa form. In field conditions, theses changes in the metabolic pattern may result from environmental PO₂ availability. In the winter, C. granulata stays in its holes, where environmental PO₂ falls to zero. The carbohydrate or protein content of the diets administrated to the crabs seem to induce different metabolic adjustments during anoxia and recovery.     

L-Lactate dehydrogenase in the longitudinal muscle of the sea cucumber Sclerodactyla briareus (Echinodermata: Holothuroidea)

The infaunal holothurian Sclerodactyla briareus (Thyone briareus) is able to tolerate exposure to hypoxic conditions for over 2 days. Since the in vitro anaerobic degradation of glucose-U-C¹⁴ by longitudinal muscle preparations leads to an accumulation of labeled lactic acid, it is apparent that lactate dehydrogenase plays a key role during anoxia. Disc electrophoresis resolved one major band of lactate dehydrogenase (LDH) activity in longitudinal muscle extracts. Muscle extracts of s. briareus oxidized L-lactate only, as is the case with most other echinoderms. The apparent Michaelis constants for lactate oxidation and pyruvate reduction are dissimilar from values reported for other marine invertebrates. The LDH activity in this holothurian is higher than the activities reported for other marine invertebrates with the exception of some active, large arthropods. Substrate inhibition of LDH maximal activity occurs at substrate concentrations (pyruvate) exceeding 10 mM, and then to only a minor extent. The characteristics of lactate dehydrogenase from the longitudinal muscle of S. briareus support the hypothesis that it functions to maintain glycolytic flux during prolonged periods of exposure to anoxic or hypoxic conditions.     

Anaerobic and aerobic energy metabolism in ovaries of the sea urchin Strongylocentrotus droebachiensis

The gonads of sea urchins undergo large changes in mass during their gametogenic cycle. In addition, they have relatively low aerobic capacities and are poorly perfused by the circulatory system and thus are continually hypoxic or anoxic. The present study of Strongylocentrotus droebachiensis investigates seasonal changes in the relationships among mass of the ovaries, pH and P_O2 of the perivisceral coelomic fluid which bathes the ovaries, and partitioning of ovary energy metabolism into its anaerobic and aerobic components. S. droebachiensis were collected at Blue Hill Falls, Maine, USA, from August 1982 to March 1984. We found that from 76 to 92% of the heat dissipated by isolated ovaries of the sea urchin S. droebachiensis derives from anaerobic energy metabolism at partial pressures of oxygen prevailing in vivo. Ovaries from S. droebachiensis have the capacity to produce large amounts of lactate under imposed anoxia, but lactate accounts for only 37% of the total anoxic heat dissipation, which suggests that other end products of anaerobiosis are present. Seasonal changes in pH and P_O2 of the perivisceral coelomic fluid can be explained by a complex relationship among changes in temperature, reproductive condition, and anaerobic and aerobic metabolism in the ovaries, gut and body wall. Seasonal changes in the buffering capacity of the perivisceral coelomic fluid must be determined before the effects of respiratory and metabolic acid production on the acid-base status of the coelomic fluid can be fully understood.     

An investigation on the anaerobic metabolism of Nephtys hombergii (Annelida: Polychaeta)

In Nephtys hombergii Savigny (Annelida: Polychaeta: Errantia), succinate and less pronounced L-alanine are accumulated as endproducts during an initial phase of anaerobiosis. In this phase aspartate is utilized as substrate for anaerobic energy production in addition to glycogen. Prolonged anaerobiosis results in the formation of propionate and acetate which, to a large extent, are excreted into the water. The concentrations of aspartate and succinate were found to remain unchanged down to a Pw_{O 2}(oxygen partial pressure in incubation water) of approximately 20 torr, indicating a fully aerobic metabolism. At a Pw_{O 2}of 15 torr characteristic changes can be observed: energy production becomes partially anaerobic. At a Pw_{O 2}of 7 torr the level of aspartate is largely reduced, but the accumulation of succinate is still significantly less than at anoxia. [In the habitat of N. hombergii, the intertidal mud flats, a Pw_{O 2}in the range from 0 torr (Pamatmat, 1968) to 12 torr (Wells and Warren, 1975) was measured.] The activities of some important enzymes involved in anaerobic energy production (among them pyruvate kinase, phosphoenolpyruvate carboxykinase and lactate dehydrogenase) were measured. Although N. hombergii has a high glycolytic capacity, no lactate dehydrogenase was detected. Instead strombine and alanopine dehydrogenases were present in comparatively high activities.     

Functional and environmental anaerobiosis in the razor-clam Ensis directus (Mollusca: Bivalvia)

In Ensis directus (Conrad) metabolic changes accompanying muscular activity and experimental anoxia were investigated. Octopine dehydrogenase, strombine dehydrogenase, alanopine dehydrogenase and lactic dehydrogenase were shown to be present in different parts of the body, but the activity of ODH always exceeded that of the other enzymes by far. Muscular activity connected with escape digging or evoked by electrical stimulation resulted in a decrease of stored phosphagen and in an accumulation of octopine. During recovery the normal concentrations of energy-rich phosphates were restored quickly, whereas octopine was removed more slowly. In experimental anoxia only minor quantities of L-and D-alanine, succinate, propionate, and acetate were formed. Quantitatively the most important endproduct was strombine, which rose linearly with the duration of anoxic incubation.     

Metabolic constraints on burst-swimming in the Antarctic teleost Notothenia neglecta

Immediate-energy-supply pathways were analysed during burst-swimming in white muscle of the Antarctic fish Notothenia neglecta, (collected during Austral summer 1984 near Signy Island, South Orkneys). Selected enzyme activities were determined, and tissue metabolites were measured, before and after 3 min of burst-work. The activity of glycolytic enzymes is very low compared to that in other fish, while the activities of creatine phosphokinase, adenylate kinase and AMPdeaminase are relatively high. Concentrations of metabolites associated with cellular energy status declined with exercise. At the same time, there was no change in the concentrations of most of the glycolytic metabolites measured (including glycogen, glucose and lactate), while [creatine phosphate] declined and [inosine monophosphate] increased. These observations were used to propose that the potential for ATP generation via creatine-phosphate hydrolysis is enhanced in N. neglecta, while glycolytic capacity is relatively reduced. This is an unusual pattern of ATP production and places severe constraints upon the behavior of the fish. While the capacity for very short-term, high-power-output swimming is increased, the capacity for medium-duration swimming is reduced. Ecological repercussions are discussed.     

Normoxic and anoxic energy metabolism of the southern oyster drillThais haemastoma during salinity acclimation

The energetic cost associated with salinity acclimation was determined in the marine gastropodThais haemastoma by direct calorimetry under normoxic and anoxic conditions. Snails were collected from Caminada Pass near Grand Isle, Louisiana (Longitude 90°2W; Latitude 29°2N) in September 1987. Metabolic heat flux of snails acclimated to and measured at 10 or 30 S was similar at 15.06 or 16.39 J g^–1 dry wt h^–1, respectively, (corresponding to 0.76 or 0.83 ml O₂ g^–1 dry flesh wt h^–1) under normoxic conditions, and 2.39 or 2.53 J g^–1 dry wt h^–1 under anoxic conditions. Inter-individual variability was high, obscuring the effect of salinity gradient on heat flux. When standardized to the pre-transfer control level of each individual under anoxic conditions, a significant increase (55%) of energy expenditure was observed for snails transferred to hyperosmotic conditions. In contrast, heat flux varied insignificantly in individuals in the anoxic 30 to 10 S transfer. After transfer of individuals from 10 to 30 S under normoxic conditions, heat flux was depressed initially to 38% of the control rate, but recovered after 14 h to a higher metabolic rate (56%) than the pre-transfer control rate. After transfer of individuals from 30 to 10 S under normoxic conditions, the standardized heat flux decreased to 28% of the control rate, followed by a 20 h period of recovery to the control rate. The energy cost of intracellular hypoosmotic regulation was less than hyperosmotic regulation under anoxic conditions. The retraction of the foot ofT. haemastoma after normoxic salinity transfers did not generally correlate with the time course of metabolic heat flux.     

Adaptation of the polychaete worm Scoloplos armiger to hypoxic conditions

The anaerobic metabolism of the intertidal polychaete Scoloplos armiger, its recovery from anaerobiosis and the importance of anaerobic energy production during low tide in the field were investigated. Under anaerobic conditions S. armiger produces energy in the same manner as Arenicola marina, a prototype of an euryoxic invertebrate from the intertidal. Energy is produced from the phosphagen stores and from the breakdown of glycogen to volatile fatty acids, mainly propionate and to a lesser extend acetate. However, S. armiger cannot reduce its energy demand to the same degree as A. marina. This and the relatively small pool of glycogen may be the reason for its only moderate resistance to anoxia. The recovery from anaerobiosis proceeds in S. armiger significantly slower than in A. marina. S. armiger is able to maintain a fully aerobic metabolism down to a Pw_{O 2}of ca. 20 torr and even at a Pw_{O 2}of 10 torr a partly aerobic metabolism was retained. In the field during low tide S. armiger ascends into the oxidative layer, where it is able to maintain an aerobic metabolism even at parts without remaining puddels on the surface.     

Anaerobic metabolism of the common cockle Cardium edule

Changes in the level of the adenylates (ATP, ADP, AMP), arginine phosphate, arginine, octopine, D-lactate, succinate, and aspartate were determined in the foot of Cardium edule, after 12 h of anoxia and after aerobic recovery for up to 12 h. Control levels of the adenylates, the calculated energy charge and the arginine phosphate content were restored after about 1 h of recovery. On the other hand, the recovery of the D-lactate, succinate, and aspartate pools was a slow process, taking place over about 12h.     

The fluffy sea anemone Metridium senile in periodically oxygen depleted surroundings

In SCUBA-diving monitored field experiments (Flensburg Fjord, 15 m, 1981) and parallel aquarium tests,the behavior of Metridium senile under anoxic conditions and its oxygen-deficiency resistance were studied. When oxygen is lacking the fluffy sea anemone diminishes body surface, and then, successively, shuts off most energy consuming activities. By means of this strategy, 50% of the tested individuals survived 3 weeks of total anoxia.