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.     

The functions of octopine dehydrogenase and D-lactate dehydrogenase in the pedal retractor muscle of thedog whelk Nassarius coronatus (Gastropoda: Nassariidae)

The pedal retractor muscle of Nassarius coronatus, unlike most mollusc muscles, contains high activities of both octopine dehydrogenase (159 IU g^-1 wet wt muscle) and D-lactate dehydrogenase (40 IU g^-1 wet wt muscle). Pedal retractor muscles show accumulation of octopine (increase of 4.7 mol g^-1 wet wt muscle), and a fall in arginine phosphate concentration (decrease of 4.1 mol g^-1 wet wt musscle) after exercise. During recovery from exercise, octopine returns to resting levels after about 120 min, D-lactate accumulates (5.1 mol g^-1 wet wt muscle), and arginine phosphate returns to resting levels within 30 min. D-lactate (2.0 mol g^-1 wet wt muscle) accumulates during long-term exposure to air. The results show that octopine dehydrogenase and D-lactate dehydrogenase can function to catalyze the terminal step of glycolysis during muscle anoxia associated with these different physiological states.     

Anaerobic metabolism in Arenicola marina and Nereis diversicolor during low tide

In order to observe the metabolic response of two marine polychaetes, Arenicola marina L. and Nereis diversicolor Müller, at various times during low tide, the concentrations of adenine nucleotides, phosphagens, D- and L-alanine, succinate, acetate, propionate and lactate were measured. During 8h of exposure ATP does not change significantly in the body-wall musculature of the lug-worm A. marina, whereas ADP and AMP concentrations double the first hour of exposure. Correspondingly, the energy charge falls from 0.91 to about 0.84. The concentration of the phosphagen phosphotaurocyamine decreases by about 20% during the first 2 h and after 6 h only about 50% of the initial concentration was measured. During exposure, aspartate decreases by approximately 6 mol g^-1 fresh weight within 6 h (from 23 to 17 mol g^-1 fresh weight). During the first 4 h of exposure succinate concentration doubles and within the next 4 h it reaches values about 10-fold the initial value. The concentrations of acetate and propionate only increase during the last 2 h (6–8) of exposure. After several hours of exposure typical changes in the concentrations of all these metabolites could also be demonstrated in N. diversicolor. The concentrations of the two phosphagens phosphoglycocyamine and phosphocreatine and of aspartate decrease by about 25% during 9 h of exposure, on the other hand succinate and especially D-lactate accumulate. In both species, the partial anaerobic energy production during exposure does not have a lasting detrimental effect. After one high tide the worms have fully recovered.     

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.     

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.     

Photosynthetic carbon dioxide fixation in zooxanthellae

The carbon-fixation patterns of freshly isolated zooxanthellae from the hermatypic coral Acropora formosa were examined during a 15 min exposure to sodium mosa were examined during a 15 min exposure to sodium [¹⁴C]bicarbonate. The labelling pattern during the first 60 s exposure showed that the C₃ carbon-fixation pathway is the major route for photosynthetic carbon fixation in Symbiodinium sp. 3-Phosphoglyceric acid, which constituted >50% of the label after 5 s, steadily decreased over the first 60 s. Hexose phosphates, aspartate, malate and glucose were the other main products during the first 60 s. Over longer periods, significant amounts of the organic acids succinate, aspartate and glutamate were found in the extract along with glucose; but no glycerol.     

Reproductive investment reduces recuperation from exhaustive escape activity in the tropical scallop Euvola zizac

In scallops, mobilization of reserves from the adductor muscle to support maintenance and reproductive activity may impinge upon a major role of the adductor muscle, the movement of the valves during swimming and escape responses. The tropical scallop Euvola ziczac (Linnaeus 1758) invests energy reserves to different degrees during its two periods of reproduction each year. We evaluated the impact of reproductive investment on the escape responses (clapping capacity and recovery after exhaustive exercise) of E. ziczac sampled at different reproductive stages (immature, mature, spawned) during the two reproductive periods in 1997. We compared the escape response capacities with the levels of muscle energetic reserves (glycogen, proteins, and arginine phosphate) and muscle metabolic capacities [activity of the glycolytic enzymes: glycogen phosphorylase (GP), pyruvate kinase (PK), phosphofructokinase (PFK), octopine dehydrogenase (ODH), arginine kinase (AK), and the mitochondrial enzyme, citrate synthase (CS)]. Gonad size, gamete volume fraction, and levels of gonadal protein and lipid were greater for mature scallops during the first than during the second reproduction. Numbers of claps during escape responses (49–57) and levels of muscle arginine phosphate remained similar throughout the different reproductive stages in both reproductive periods. In contrast, recovery was slowed during gonadal maturation in both reproductive periods and during spawning in the first reproduction. Scallops generally took more time to regain their initial clapping capacity during the first (25–40 min) than during the second reproduction (20–30 min). Muscle glycogen decreased markedly during both gonadal maturation and spawning in both reproductions; whereas, muscle proteins decreased only in the first reproduction. The levels of most enzymes decreased during gonadal maturation in both reproductions, and also after spawning, particularly during the first reproduction. We concluded that gonadal maturation and spawning did not decrease clapping capacity of E. ziczac, but decreased its capacity to recover from exhaustive exercise most likely due to decreased levels of energetic reserves and a reduced metabolic capacity of the adductor muscle. Moreover, these effects were probably stronger during the first cycle because of the greater reproductive investment coincident with decreased food availability. Received: 28 April 2000 / Accepted: 21 September 2000     

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.     

Quantification of arginine requirements of juvenile marine shrimp,Penaeus monodon,using microencapsulated arginine

Using microencapsulated L-arginine, the quantitative requirement for amino acid has been determined for the first time for a shrimp species, which can not effectively utilize crystalline amino acids. In an 8 wk feeding trial (1990), juvenile Penaeus monodon were fed casein-based purified diets containing one of six levels (13.1, 17.7, 22.3, 26.9, 31.5 and 36.1 g/kg diet) of arginine. In addition to the protein-bound arginine already present in the casein of the test diets, pure arginine was supplemented by L-arginine microencapsulated in cellulose acetate phthalate, which is easily assimilated. The arginine level required for optimal growth was determined by brokenline model analysis of weight gain data to be 25.0 g/kg diet (=54.7 g/kg protein). Examination of the hemolymph 3 h after feeding revealed that the free arginine level in the hemolymph had not increased as a result of increasing levels of the dietary arginine. There was an abrupt increase of urea in the hemolymph when the arginine requirement of the shrimp had been met.     

Effect of freezing on photosynthesis of intertidal macroalgae: relative tolerance of Chondrus crispus and Mastocarpus stellatus (Rhodophyta)

The effect of freezing on photosynthetic metabolism was studied in the red algae, Chondrus crispus and Mastocarpus stellatus. Plants of both species were collected from the intertidal at Chamberlain or Kresge Point, Maine, USA (43°56N, 69°54W) between February and March 1987. Photosynthetic rates were measured immediately after freezing at-20°C and following recovery periods in seawater. Photosynthesis in C. crispus declined rapidly following freezing, falling to 70% of control values within 1 h and 30% after 3 h exposure. Minimum photosynthetic rates (7 to 9% of controls) occurred following freezing exposures of 12 h or more. Full photosynthetic recovery in C. crispus after 3 h at-20°C required 48 h. Photosynthesis in C. crispus did not fully recover in plants frozen for 6 h or more. In contrast, photosynthesis in M. stellatus was relatively unaffected by freezing exposures of <12 h. Twelve hours or more at-20°C reduced photosynthesis to 55% of controls. Photosynthesis in M. stellatus fully recovered from 24 h at-20°C within 24 h. In both species the reduction of photosynthesis by freezing was associated with damage to the plasma membrane and reduced efficiency of energy transfer from phycobilisomes to chlorophyll a, but did not appear to involve ribulose-1,5-bisphosphate carboxylase oxygenase activity. The freezing tolerance of C. crispus and M. stellatus positively correlates with their respective intertidal distributions, suggesting that freezing may be involved in controlling the distributions of these species on the shore.     

Photoinhibition and recovery of the kelp Laminaria saccharina at optimal and superoptimal temperatures

Effects of high irradiance on photosynthetic characteristics were examined in sporophytes of the kelp Laminaria saccharina Lamour. from 1992 to 1994. Exposure to high irradiance (700 mol photons m^-2s^-1) for 1 h at optimal temperature (12°C) caused a 40 to 60% decline in photosynthetic efficiency (alpha), quantum yield, and the ratio of variable to maximum chlorophyll fluorescence (F_v/F_m), an indicator of Photosystem II efficiency. Although the photoinhibition effects were partly attributable to protective mechanisms, a concurrent increase in minimal fluorescence (F_o) indicated damage to Photosystem II reaction centers. The magnitude of photoinhibition was proportional to irradiance and duration; however, F_v/F_m was significantly reduced after exposure to irradiances as low as 40 to 50 mol photons m^-2s^-1 for 1 h, or to 700 mol photons m^-2s^-1 for only 5 min. In contrast, photosynthetic capacity (P_max) was affected only at much higher irradiance. Superoptimal temperatures up to 24°C did not exacerbate high-light effects. At 25°C, however, alpha and P_max were more susceptible to photoinhibition than at lower temperatures. Recovery from photoinhibition was examined by following F_v/F_m and F_o for 24 h after exposure to high light. Recovery of F_v/F_m was fastest during the first 1 to 3 h, and slowed or ceased after 6 to 12 h, while recovery of F_o was relatively constant over 12 h. Dithiothreitol, which blocks formation of energy-dissipating xanthophylls, reduced both the initial rate and extent of recovery. Chloramphenicol, which blocks chloroplast-encoded protein synthesis, had little effect on initial rates of recovery, but stopped recovery after 3 h. Thus, L. saccharina appears to rely on the xanthophyll cycle to protect the photosynthetic apparatus, and reversal of this protective mechanism causes the rapid initial recovery in F_v/F_m. Longterm recovery depends on repair of damaged reaction centers. Both the rate and extent of recovery were temperature-dependent. The initial rate was higher at 18 to 22°C than at 12°C, but the extent of recovery over 24 h declined with increasing temperature. High temperatures, therefore, appear to enhance protective mechanisms, but disrupt repair processes. L. saccharina from Long Island Sound, an ecotype adapted to low light and high temperature, showed slightly but consistently greater effects of photoinhibition than plants from the Atlantic coast of Maine, but exhibited faster recovery at superoptimal temperatures.     

Survival of the intertidal pulmonate Onchidium tumidium during short term and long term anoxic stress

Onchidium tumidium showed a triphasic response to anoxia. Twelve hours of anoxic exposure had no effect on the glycogen content in O. tumidium. However, there were significant increases in the alanine, lactate and succinate contents in the anoxic individuals. These were accompanied by a significant decrease in the ATP content. These results suggest that O. tumidium survived the first 12 h of anoxic exposure without increasing the glycolytic flux to compensate for the lower efficiency of ATP production through anaerobic pathways. Indeed, the fructose-2,6-bisphosphate (F-2,6-P₂) content and the percentage of phosphofruc-tokinase (PFK) associated with the subcellular particles remain unchanged in O. tumidium exposed to 12 h of anoxia. Hence, a reduction in the metabolic rate of these individuals might have occurred during such a period of anoxia. In contrast, in between 12 and 24 h of anoxic exposure, the glycogen content O. tumidium decreased significantly, and levelled off thereafter. A significant increase in the percentage of PFK associated with the subcellular particles was observed in individuals exposed to 24 h of anoxia. In addition, the F-2,6-P₂ content of these anoxic individuals increased significantly. Taken together, these two mechanisms could activate PFK and lead to a greater glycolytic flux. Beyond 24 h of anoxic exposure, survival of O. tumidium must have required considerable suppression of metabolism as accumulation of end products and depletions of glycogen and ATP had reached constant levels.     

Sub-lethal effect of chlorpyrifos on protein metabolism of the food fish Clarias batrachus and monitoring of recovery

The effect of sub-lethal concentrations of chlorpyrifos on protein metabolism in physiological important tissues, namely gills, kidney, liver, and muscle of the freshwater fish, Clarias batrachus, was studied. Fish were exposed to 1/20th and 1/10th of 96?h LC₅₀ concentrations for 7, 14, 21, and 28 days. After 28 days of exposure, fish were released into fresh water and kept in the same for 21 days in order to study the recovery. Fish were sacrificed at the stipulated periods and gills, kidney, liver, and muscle tissues were used for the estimation of total protein, amino acids, ammonia, urea, glutamine, protease, transaminases, and phosphatases. Total protein, amino acid, and ammonia contents were decreased in all tissues for 28 days and recovery was observed during the recovery period. Urea and glutamine levels were elevated, except in kidneys, and recovered at the end of the recovery period. The activities of protease, alanine, and aspartate aminotransferases, and acid and alkaline phosphatases were elevated in the tissues for 28 days exposure at both concentrations. Recovery of these enzymes activities was noticed during depuration.     

Nutrient stimulation of spore settlement in the kelpsPterygophora californica andMacrocystis pyrifera

FertilePterygophora californica Rupr. andMacrocystis pyrifera (L.) C.Ag. were collected in California, USA, from 1987 to 1989. Settlement activity of the spores was stimulated by nutrients, but this was not constant over time. Nutrients had no effect on the settlement ofP. californica spores between 2 and 18 h after sunrise, but settlement activity was stimulated by a nutrient mixture between 20 and 24 h after sunrise (14 to 18 h after release). Settlement activity inM. pyrifera spores was unaffected at 2 to 3 h after release, but settlement activity was significantly stimulated from 5 to 12 h after release. A variety of individual nutrients significantly stimulated settlement in 8 to 9 h oldM. pyrifera spores: ammonium, nitrate, glycine, phosphate, manganese-EDTA (18µM), borate, ferrous iron-EDTA, and ferric iron-EDTA. Spores also settled readily in unenriched artificial seawater, and nutrient-stimulated settlement rates were usually 150% of the unenriched control levels. Neither EDTA alone, cobalt-EDTA, nor manganese-EDTA (2µM) had significant effects onM. pyrifera spore settlement. The effects of time and of several individual nutrients on spore settlement activity are different from previously reported chemotactic effects of nutrients onP. californica andM. pyrifera spores. It is suggested that nutrient settlement-stimulation is mechanistically different from nutrient chemotaxis. However, like chemotaxis, settlement stimulation is probably an adaptation which increases the likelihood of spore settlement in microhabitats suitable for subsequent growth and reproduction of gametophytes.     

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     

The enzymes involved in synthesis and utilization of carbamylphosphate in the deep-sea tube worm Riftia pachyptila

 The obligate symbiosis of the deep-sea tube worm Riftia pachyptila with a sulphur-oxidizing bacterium raises important questions concerning its metabolism and metabolic exchanges. In this study, the presence and properties of the enzymes synthesizing and utilizing carbamylphosphate in the arginine and pyrimidine nucleotide pathways were investigated in this worm. The results show that the ammonium-dependent carbamylphosphate synthetase and ornithine transcarbamylase, enzymes involved in the arginine pathway, are present in all body parts of the worm. In contrast, the glutamine-dependent carbamylphosphate synthetase and aspartate transcarbamylase, enzymes involved in the de novo pathway for pyrimidine nucleotides biosynthesis, are present only in the trophosome, the symbiont-harbouring tissue. Although the bacterial nature of these enzymes is not unambigously established, these results strongly suggest that the de novo biosynthesis of pyrimidine nucleotides is limited to the trophosome, the organ where the production of metabolic energy takes place, while the other parts of the worm's body rely on the salvage pathway for the production of the pyrimidine triphosphate nucleotides. Received: 22 June 1998 / Accepted: 26 October 1999     

Metabolic responses in zebrafish (Danio rerio) exposed to zinc and cadmium by nuclear magnetic resonance -based metabolomics

Heavy metals are common marine and soil pollutants that are mainly the result of industrial activity, and are a threat to the environment and human health. In this study, ¹H nuclear magnetic resonance (NMR)-based metabolomics was applied to adult Danio rerio to monitor the metabolic change as a response to ZnCl₂ and CdCl₂ exposure at different concentrations for 72?h. NMR spectroscopy was used to identify and quantify the metabolites extracted from D. rerio. The metabolite profiles of the control and heavy metal exposed group were classified by partial least squares – discriminant analysis (PLS-DA) analysis, and potential contaminant-specific biomarkers were suggested. For the ZnCl₂-exposed zebrafish, the levels of ATP, aspartate and NAD⁺ were increased, whereas the levels of formate, inosine, hypoxanthine and succinate decreased. In addition, the CdCl₂-exposed zebrafish showed an increase in the levels of ATP and formate and a decrease in the levels of glutamate, inosine and glutathione. Overall, Zn and Cd may lead to neurotoxicity, disturbances in the energy metabolism and oxidative stress. Our finding demonstrated that the application of NMR-based metabolomics might be useful for detecting the toxicity caused by sub-lethal concentrations of heavy metal contaminants in the environment.     

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.     

Drought stress,rain and recovery of the intertidal seaweed <Emphasis Type="Italic">Fucus spiralis</Emphasis>

Non-motile organisms of intertidal shores such as seaweeds have to cope with a great variability of environmental factors. In this survey, we studied whether different morphotypes of the intertidal seaweed Fucus spiralis L. are also reflected in a characteristic performance. Desiccation and recovery of this Phaeophyceae were investigated in field experiments near Aljezur, Portugal. Fucus spiralis is exposed to serious desiccation during periods of falling tide, resulting in a tissue water loss of about 90%. Due to large semidiurnal tidal ranges in this area, two morphotypes can be distinguished: F. spiralis growing in the lower intertidal (LZ) is thicker and fleshier compared with plants in the upper intertidal (HZ), and this is reflected in a significant difference in fresh and dry mass. During sunny days and at low tide, effective quantum yields (Φ_PSII) decreased significantly after 2 h desiccation. This continued until re-submersion. The photosynthetic performances of HZ and LZ plants also differed significantly after LZ plants were already submerged and photosynthetisizing, but the HZ specimens still exposed to air. Recovery experiments after desiccation treatments showed fast recovery within 6 min after re-submersion in both morphotypes. HZ specimens showed a slower recovery, which indicates a protection measure to the adverse conditions in the upper intertidal. In 24 h desiccation treatments, however, HZ specimens expressed a significantly higher maximum fluorescence yield F _v /F _m recovery. Simulated rainfalls during low tides caused photosynthetic activity to drop to 50% of initial F _v /F _m , independent of the length of the rain period. Treated plants also fully recovered after 6 min re-submersion in seawater. A comparison of single fronds and tufts clearly indicated advantages of the tuft growth strategy: tufts showed higher Φ_PSII at prolonged emersion times. Our study indicated a clear relationship between size and drought resistance, which was primarily due to the smaller and hardy HZ plants that withstand longer desiccation times without damage.