Effects of temperature and food on the free amino acids in tissues of roach (Rutilus rutilus L.) and rudd (Scardinius erythrophthalmus L.)

• 1.1. In liver and muscle the concentrations of free amino acids (FAA) are highest in fish maintained at low temperature and fed mealworms. These effects are more pronounced in roach than in rudd.
• 2.2. In the liver alanine, glycine and glutamate are the dominant FAA but proline increases in mealworm-fed animals.
• 3.3. In muscle, histidine and glycine dominate, except that a mealworm diet leads to an increase in the concentration of proline and to a concomitant decrease in the concentration of glycine.
• 4.4. Starvation leads to a reduction of total FAA content but to relative increases of lysine and histidine. These two FAA can serve as indicators of the general state of nutrition of roach and rudd.
• 5.5. The molar ratio [gly]/[his] is strongly correlated with temperature, decreasing with an increase in the temperature to which the animals had been exposed prior to capture.
• 6.6. The patterns of free and bound amino acids diverge more widely in these species than in mammals which reflects the greater dependence of the FAA pools of fish on intrinsic and extrinsic factors.
• 7.7. The concentrations of histidine in the FAA pools of muscle and in food proteins are strongly correlated.

     

Effect of metabolic inhibitors on hyperosmotically induced free amino acid accumulation in the isolated foot muscle of the brackish-water bivalve Corbicula japonica

• 1.1. Accumulation of free amino acids (FAA) in the isolated foot muscle of the brackish-water bivalve Corbicula japonica during the initial stage of hyperosmotic stress was quantitatively and qualitatively similar to that in the foot of the intact animal.
• 2.2. Aminooxyacetate (AOA), a transaminase inhibitor, markedly inhibited alanine accumulation and promoted ornithine accumulation in the isolated foot. Iodoacetate (IAA), a glycolytic inhibitor, caused no significant alteration in the alanine level and the TLC pattern of FAA. Both the inhibitors scarcely influenced the pool size of total ninhydrin positive substances (NPS).
• 3.3. A major part of the carbon of accumulated FAA during the initial stage of hyperosmotic stess did not seem to arise from glycolysis.
• 4.4. Free d-alanine as well as l-alanine accumulated in isolated foot muscle exposed to hyperosmotic stress.

     

Time related changes in the free amino acid pool of the sea anemone,Bunodosoma cavernata,during salinity stress

• 1.1. The sea anemone, Bunodosoma cavernata, is a relatively eurybaline cnidarian tolerating salinities from 12 to 40%.
• 2.2. Taurine, glutamic acid and aspartic acid all showed some increases with increased salinity.
• 3.3. The amino acid showing the greatest accumulation under high salinity conditions was β-alanine which increased 28-fold from 1.5 to 41.9 μmol/g dry weight when salinity was raised from 26 to 40%.
• 4.4. When B. cavernata was subjected to increased salinity, β-alanine was rapidly accumulated and reached maximum levels within 4 days.
• 5.5. When salinity was dropped from 36 to 26%0, β-alanine concentrations dropped from 15 to 2 μmol/g dry weight in 2 days.

     

Amino acid synthesis during hyperosmotic stress in penaeus aztecus postlarvae

• 1.1. Glycine, proline, and taurine are the quantitatively most important amino acid osmolytes in Penaeus aztecus postlarvae.
• 2.2. Taurine dominates the amino acid pool in low salinity, while proline dominates the amino acid pool at higher salinities.
• 3.3. Although not major contributors to the pool, glutamate and alanine are constitutively synthesized from [¹⁴C]glucose and [¹⁴C]glutamate under constant salinity and under hyperosmotic stress treatments.
• 4.4. Proline synthesis from [¹⁴C]-precursors is apparent under constant high (but not low) salinity and is significantly induced by hyperosmotic stress.
• 5.5. No appreciable glycine synthesis was observed from [¹⁴C]glucose or [¹⁴C]glutamate under any experimental conditions.

     

Changes in chemical composition of muscle in young hybrids between russian sturgeon Acipenser guldenstadti brandt × beluga Huso huso l. (Pisces; acipenseriformes) under different levels of salinity

• 1.1. The salinity tolerance in young RS × B hybrids increases as the fingerlings grow. The specimens weighing about 7 g are able to tolerate the direct transfer to the water salinity 18%..
• 2.2. Under hypo- and iso-osmotic water ion concentration in the hybrid muscle free amino acids, the exchange of taurine for β-alanine and glycine takes place.
• 3.3. Under hyperosmotic conditions within the first 2 days in the hybrid muscle the water quantity declines, the protein quantity also slightly decreases, the urea and free amino acids concentration (mostly alanine, aspartic and glutamic acids, leucine), and a portion of reserved lipids increase.
• 4.4. During the next 4 days the muscle moisture, protein quantity, and the concentration of urea and free amino acids return to control values, but the portion of reserved lipids declines below the original level.

     

Effects of anoxia on the haemolymph physiology and lactate concentrations in the freshwater crab Potamon warreni calman

• 1.1. Haemolymph lactate levels rose rapidly from 0.54 ( ± 0.39) mmol to 34.78 ( ± 4.9) mmol during 6 hr of anoxia in a N₂ atmosphere.
• 2.2. A sharp decrease in the pH from 7.478 (± 0.04) to 7.197 ( ± 0.04) and the total carbon dioxide content of the haemolymph from 13.97 (± 2.0) mmol to 6.25 (± 1.2) mmol during anoxia indicates a gross disturbance in the acid-base balance in Potamon warreni.
• 3.3. The low concentrations of succinate (98 ± 30 μmol) and alanine (5.8 ± 1.0 mmol) in the haemolymph suggest that they do not play a role as an energy source during anoxia.
• 4.4. Probably only l-( + )-lactate is produced during lactate production when P. warreni is exposed to anoxic conditions.

     

The influence of seasonal changes on energy metabolism in Mytilus edulise (L.).—III. Anaerobic energy metabolism

• 1.1. Seasonal changes in the accumulation of end products after 48 hr of exposure to air and in the composition of the free amino acid pool were studied in Mytilus edulis.
• 2.2. The accumulation levels of succinate and acetate showed only weak seasonal changes.
• 3.3. Conversion of succinate to propionate was high in summer and virtually zero in winter
• 4.4. Alanine and most other free amino acids were present in relatively high concentrations in summer and early autumn and reached minimal values in winter and early spring.
• 5.5. Exceptions were glutamate, aspartate and taurine, which showed hardly an season related changes and glycine, which changed inversely to the majority of the free amino acids.
• 6.6. The anaerobic formation of alanine was inversely proportional to the endogenous concentration.
• 7.7. The only other free amino acids affected by anaerobiosis were glutamate and aspartate, which respectively increased and decreased under these conditions.

     

Aerobic glucose disposal in the oyster: An in vivo kinetic analysis

• 1.1. Aerobic glucose disposal in starved oysters exposed to 1 mM external glucose was 2.29 μg C/g wet wt/min.
• 2.2. It was hypothesized that the maximum disposal rate is limited by the maximum rate of transepithelial glucose transport.
• 3.3. The major recipients of glucose-carbon were glycogen and amino acids. 4. The rate of glucose-carbon disposal to these two pools was 0.80 and 0.42 μg C/g/min, respectively.
• 4.5. The internal energy state determines the pathways of glucose disposal.
• 5.6. Disposal of glucose-carbon in “glucose-primed” oysters is primarily into glycogen.
• 6.7. In fasted bivalves the disposal is primarily into amino acids and carboxylic acids.
• 7.8. The uptake of dissolved glucose has the potential of contributing significantly to growth under conditions where the external glucose concentration is kept artificially high.

     

Anaerobic metabolism in sublittoral living Mytilus galloprovincialis in the mediterranean—IV. Role of amino acids in adaptation to low salinities during anaerobiosis and aerobiosis

• 1.1. When Mytilus galloprovincialis were transferred from 38 to 19%. sea water (S), the metabolism became anaerobic for at least 8 hr. After 24 hr the animals were entirely aerobic again.
• 2.2. Upon transfer to 19%. S, the total free amino acid concentration in haemolymph doubled within 4 hr, remaining nearly constant thereafter, up to 48 hr.
• 3.3. In the posterior adductor muscle a strong decrease of alanine and glycine occurred at 48 hr exposure to 19%. S, and a smaller decrease of glutamate; taurine remained relatively constant. When transferred again to 38%. S after 14 days, a strong overcompensation occurred in the concentrations of alanine and proline, and a smaller overcompensation in those of threonine and serine.
• 4.4. In the gill no distinct change in the amino acid pool occurred during 14 days of exposure, with the exception of a decrease in serine. When transferred again to 38%. S, a strong overcompensation occurred in alanine, proline, glycine and serine, and a smaller in glutamate and threonine.
• 5.5. No evidence for anaerobic metabolism in the decrease of the amino acid pool was found.
• 6.6. M. galloprovincialis is less able to adapt to low salinities than the more euryhaline M. edulis.

     

A comparison of aspartate transcarbamylase activity in juvenile pacific oysters (Crassostrea gigas) when fed various diets

• 1.1. The optimum pH for measurement of aspartate transcarbamylase activity in oyster tissue was determined to be 9.35 while the optimum temperature was 39.5°C.
• 2.2. Aspartate transcarbamylase activity varied significantly over short periods of time (hr) possibly due to fluctuations in the amount of food digested.
• 3.3. The composition of the oyster's diet also affected the levels of aspartate transcarbamylase activity in oyster tissues.
• 4.4. Those oysters fed an egg yolk-starch diet contained significantly lower aspartate transcarbamylase activity than oysters fed an egg yolk-starch-salmon oil diet or a casein-starch-salmon oil diet.
• 5.5. The aspartate transcarbamylase activities in oysters fed Phacedactylum tricornutum or a starch diet were not significantly different from the activities in oysters fed the egg yolk-starch diet.

     

Changes in free amino acid composition in haemolymph of larvae of the wax moth,Galleria mellon ella L., during cold acclimation

• 1.1. Free amino acids were analysed in the haemolymph of Galleria mellonella larvae by HPLC chromatography with o-phthaldialdehyde (OPA)-l-thio-β-d-glucose as derivatization agent.
• 2.2. Fourteen primary amino acids were detected among which glutamine, alanine, γ-aminobutyric acid (GABA) and glycine predominated and constituted 67.7% of the amino acids found.
• 3.3. The concentration of GABA increased significantly with the age of larvae entering the wandering phase and reached a maximum during metamorphosis.
• 4.4. Analysis of cold-acclimated larvae revealed a net increase of free primary amino acids from 96 to 151.8 μmol/ml during consecutive acclimation to 0°C within 20 days and to 205.4μmol/ml during cold shock injury at 0°C (3 hr).
• 5.5. The bulk of this increase was accounted for by alanine, glycine, phenylalanine and lysine.

     

The effect of changes in external salinity on the free amino acids and two aminotransferases of white muscle from fasted Salmo gairdneri Richardson

• 1.1. Aspartic acid. glutamic acid and serine concentrations in the white muscle of starved rainbow trout kept in diluted sea water (600 mOsm/l) for 8 days were significantly higher than in control animals kept in fresh water.
• 2.2. After 24 days the levels of all amino acids investigated (aspartic acid, glutamic acid, serine, glycine. alanine, threonine and lysine) in the white muscle of starved rainbow trout kept in diluted sea water were higher than in the white muscle of animals kept in fresh water without food.
• 3.3. Alanine aminotransferase activity in starved rainbow trout kept in diluted sea water for 24 days was higher than in the control animals kept in fresh water.
• 4.4. There is a significant correlation between alanine concentration and alanine aminotransferase activity in the white muscle of rainbow trout.

     

The effect of high salinity on development,mortality and ray number of Echinaster spinulosus (Echinodermata: Asteroidea) at different developmental stages

• 1.1. Embryos and larvae of the asteroid Echinaster spinulosus were exposed to high salinity stress at various stages during early development.
• 2.2. Highest percentages of three-rayed (9.7%) and four-rayed (29%) individuals occurred when individuals which had developed for 48 hr (appearance of pre-oral lobe) at ambient salinity (30%o) were exposed to high salinity (39%o).
• 3.3. The percentage of ray-number aberrancies increased with increasing salinity.
• 4.4. The ontogenetic events associated with the formation of the hydrocoelic rudiments at the pre-oral lobe stage may be sensitive to salinity and influence the development of ray number.
• 5.5. The ability to induce variations of ray number in asteroids with salinity stress may yield an experimental approach for the determination of the adaptive significance of ray number in asteroids.

     

Organ specific changes in energy metabolism due to anaerobiosis in the sea mussel Mytilus edulis (L.)

• 1.1. Anaerobic energy metabolism was investigated in different organs of Mytilus edulis and the whole animal.
• 2.2. Succinate accumulates to high levels in most organs but remains low in the hemolymph.
• 3.3. After 16 hours propionate accumulation is observed in all organs. Experimental evidence is not sufficient yet to point out organs that produce more propionate than others.
• 4.4. Acetate is a minor end product.
• 5.5. Acetate and propionate are found in the hemolymph in amounts equal to those in the organs.
• 6.6. Animals incubated in oxygen-free seawater accumulate more end products than animals exposed to air, in the form of volatile fatty acids that are excreted into the incubation water.
• 7.7. Alanine and glutamine increase in the posterior adductor muscle. Aspartate decreases in the total animal, posterior adductor muscle and gills, while in the hemolymph decrease in alanine, asparagine, serine, threonine and proline are observed.

     

Variations in response to environmental hypoxia of different colour forms of the shore crab,Carcinus maenas

• 1.1. The oxygen consumption of red and green Carcinus in normoxic and hypoxic sea water was determined, using an oxygen electrode in a sealed respirometer.
• 2.2. The red crabs had significantly higher “excited” oxygen uptake rates and a lower ability to compensate for hypoxia than the green crabs.
• 3.3. Red Carcinus display an emersion response to declining oxygen at lower oxygen tensions than the green crabs.
• 4.4. Mortality of red crabs exposed to prolonged anoxia was much greater.
• 5.5. The relationship of these findings to the zonation of the two colour forms on the shore is discussed.

     

Metabolite concentrations in electrically stimulated lugworms Arenicola marina

• 1.1. The concentrations of lactate, succinate, alanine, aspartate, acetate and propionate in the lugworm Arenicola marina were measured after 1, 2.5 and 12 hr of continuous electrical stimulation under aerobic and anaerobic conditions.
• 2.2. A continuous increase of the concentrations of alanine and acetate, and a decrease of aspartate occurred during the first 2.5 hr of electrical stimulation. A marked rise of succinate and propionate was observed only in experiments lasting longer than 2.5 hr.
• 3.3. No changes were detected in the concentrations of lactate. Under anoxia the metabolites accumulated at significantly higher rates than under aerobic conditions.

     

Proteolysis post mortem in North Atlantic krill

• 1.1. The autoproteolytic processes in selected species of North Atlantic krill, Meganyctiphanes norvegica (M. Sars), Thysanoessa inermis (Krøyer) and T. raschii (M. Sars) have been examined at 0°C by following the release of peptides and free amino acids.
• 2.2. The krill contains high levels of peptide hydrolases, and autoproteolysis seems to be due mainly to digestive enzymes localized in the hepatopancreas and the intestinal tract of the animals.
• 3.3. During autoproteolysis the individual amino acids were generally released at rates corresponding to their proportion in the bulk protein of the krill. The major exceptions were alanine which accumulated in amounts larger than was to be expected from the composition of the krill protein, and glutamic acid/glutamine, aspartic acid/asparagine, arginine, and to some extent glycine, proline and serine, which accumulated to a lesser extent than was to be expected.
• 4.4. Storage of krill for 1 week resulted in only minor changes in the total content of amino acids as determined after acid hydrolysis, with the exception of alanine which increased in concentration.
• 5.5. The results suggest that the formation of free alanine is partly due to reactions other than proteolysis.
• 6.6. The release of free amino acids was accompanied by a considerable increase in the amount of small peptides, and glutamic acid/glutamine, aspartic acid/asparagine, glycine and proline tended to accumulate in these peptides.
• 7.7. The autoproteolytic activity of the Thysanoessa species showed seasonal variations, probably in response to food availability. In the case M. norvegica, the results suggest that there are smaller fluctuations in the level of proteolytic enzymes, probably indicating less pronounced variations in the food intake over the year.

     

Anaerobic metabolism of the common cockle,Cardium edule—IV. Time dependent changes of metabolites in the foot and gill tissue induced by anoxia and electrical stimulation

• 1.1. The levels of adenylates, arginine phosphate, arginine and various other metabolites related to anaerobic metabolism were determined in foot and gills of Cardium edule during anoxia (incubation in oxygen-free sea water) up to 48 hr and after electrical stimulation of the foot muscle.
• 2.2. In the foot the phosphagen was depleted quite rapidly in the first hours of anoxia. Free arginine levels rose while there was little octopine accumulation. d-Lactate appeared to be the predominant end product. Alanine and succinate were also produced and the concentration of aspartate decreased, suggesting this metabolite as precursor for succinate. In the gills, where no arginine phosphate was present, succinate production was higher than in the foot and exceeded lactate formation.
• 3.3. Only a small decline in ATP occurred during anoxia and consequently, only moderate changes in the energy charge were observed in both tissues.
• 4.4. Electrical stimulation of the foot also resulted in the breakdown of arginine phosphate and formation of mainly lactate and not octopine.
• 5.5. Total ATP consumption during anoxia has been calculated to be more rapid during the first hours. The ATP consumption rate was reduced by a factor of 7.8 during 48 hr of anoxia.
• 6.6. During the first 4 hr of anoxia about 60% of the total consumed energy (as ATP equivalents) was provided by the phosphagen, while later glycolysis contributed about 80%. During electrical stimulation about 75% of the energy supplied was derived from the phosphagen.
• 7.7. A comparison of the C. edule results with the data during anoxia and exercise of the jumping cockle is made.

     

The free amino acids of muscle in the cephalochordate Branchiostoma,the cyclostome Myxine and some fishes

• 1.1. Branchiostoma and Myxine have the highest concentrations of amino acids (207 and 234 mM) of the five species investigated.
• 2.2. The predominant amino acids are glycine, proline, alanine, taurine, serine and valine, which form 83–98% of the total, except in Latimeria (60%).
• 3.3. Total amino acids are considered from the point of view of osmotic concentration in relation to other nitrogenous compounds of muscle.

     

Chemical composition of North Atlantic krill

• 1.1. The main chemical components of Meganyctiphanes norvegica (M. Sars), Thysanoessa inermis (Krøyer) and T. raschii (M. Sars) have been examined.
• 2.2. Protein accounted for 42–47% of the dry weight of M. norvegica and 32–50% of the dry weight of the Thysanoessa species. On a wet weight basis, the protein content was relatively constant and independent of season.
• 3.3. The dominating amino acids in the bulk protein of the krill were glutamic acid/glutamine, aspartic acid/asparagine, glycine, alanine, lysine and leucine.
• 4.4. Lipids were present in amounts of 13–29% of the dry weight in M. norvegica, 15–50% in T. inermis and 12–44% in T. raschii, and the lipid content varied with season.
• 5.5. The main nitrogen extractives in krill, expressed on a dry weight basis, were free amino acids (5–10%), trimethylamine oxide (about 4%), peptides (about 1%) and nucleotides (0.4–1.3%). Trimethylamine and ammonia were present in very low concentrations in living krill.
• 6.6. The amino acids taurine, glycine, proline, arginine, sarcosine and alanine made up 89–93 mol% of the free amino acid pool.
• 7.7. The ash content of krill was in the order of 10–13% of the dry weight, and fluoride represented 1040 and 3200 ppm in the Thysanoessa species and M. norvegioca, respectively.