Protein synthesis and specific dynamic action in crustaceans: effects of temperature

Temperature influences the specific dynamic action (SDA), or rise in oxygen uptake rate after feeding, in eurythermal and stenothermal crustaceans by changing the timing and the magnitude of the response. Intra-specific studies on the eurythermal crab, Carcinus maenas, show that a reduction in acclimation temperature is associated with a decrease in SDA magnitude, resulting from an increase in SDA duration but a decrease in peak factorial scope (the factorial rise in peak SDA over prefeeding values). Inter-specific feeding studies on stenothermal polar isopods revealed marked differences in SDA response between the Antarctic species, Glyptonotus antarcticus and the Arctic species, Saduria entomon. Compared to S. entomon held at 4 and 13 degrees C, the SDA response in G. antarcticus held at 1 degrees C was characterised by a lower absolute oxygen uptake rate at peak SDA and an extended SDA duration. At peak SDA, whole animal rates of protein synthesis increased in proportion to the postprandial increase in oxygen uptake rate in the Antarctic and the Arctic species. Rates of oxygen uptake plotted against whole animal rates of protein synthesis gave similar relationships in both isopod species, indicating similar costs of protein synthesis after a meal, despite their differences in SDA response and thermal habitat.     

Effect of protein synthesis inhibitor cycloheximide on starvation,fasting and feeding oxygen consumption in juvenile spiny lobster Sagmariasus verreauxi

Metabolism in aquatic ectotherms evaluated by oxygen consumption rates reflects energetic costs including those associated with protein synthesis. Metabolism is influenced by nutritional status governed by feeding, nutrient intake and quality, and time without food. However, little is understood about contribution of protein synthesis to crustacean energy metabolism. This study is the first using a protein synthesis inhibitor cycloheximide to research contribution of cycloheximide-sensitive protein synthesis to decapod crustacean metabolism. Juvenile Sagmariasus verreauxi were subject to five treatments: 2-day fasted lobsters sham injected with saline; 2-day fasted lobsters injected with cycloheximide; 10-day starved lobsters injected with cycloheximide; post-prandial lobsters fed with squid Nototodarus sloanii with no further treatment; and post-prandial lobsters injected with cycloheximide. Standard and routine metabolic rates in starved lobsters were reduced by 32% and 41%, respectively, compared to fasted lobsters, demonstrating metabolic downregulation with starvation. Oxygen consumption rates of fasted and starved lobsters following cycloheximide injection were reduced by 29% and 13%, respectively, demonstrating protein synthesis represents only a minor component of energy metabolism in unfed lobsters. Oxygen consumption rate of fed lobsters was reduced by 96% following cycloheximide injection, demonstrating protein synthesis in decapods contributes a major proportion of specific dynamic action (SDA). SDA in decapods is predominantly a post-absorptive process likely related to somatic growth. This work extends previously limited knowledge on contribution of protein synthesis to crustacean metabolism, which is crucial to explore the relationship between nutritional status and diet quality and how this will affect growth potential in aquaculture species.

     

Relationship between specific dynamic action and protein deposition in calanoid copepods

The link between specific dynamic action (SDA) and protein deposition was investigated in copepodites stage V of two calanoid copepod species, the neritic Acartia tonsa and the oceanic Calanus finmarchicus. This was done by measuring respiration before, during, and after a specific feeding period and measuring the incorporation of carbon into proteins. These were also measured on individuals incubated with cycloheximide, an antibiotic that inhibits protein synthesis. The cycloheximide treatment significantly diminished the magnitude of SDA in both A. tonsa and C. finmarchicus, and inhibited carbon incorporation into protein in both species. This provides evidence that the rate at which protein deposition takes place greatly affects the magnitude of SDA. The specific respiration rates of both starving and feeding copepods were generally higher in A. tonsa than in C. finmarchicus. This influenced SDA, the magnitude of SDA normalised to an 8 h feeding period being threefold higher in A. tonsa (78.7+/-25.7 nlO(2) μgC(-1)) than in C. finmarchicus (27.5+/-11.6 nlO(2) μgC(-1)). This difference may arise due to differences in energy allocation in the organisms of the copepodite V stage of the two species. In this stage C. finmarchicus deposits large quantities of storage lipids, predominately wax esters, whereas A. tonsa deposits proteins during somatic growth.     

The effect of meal composition on specific dynamic action in burmese pythons (Python molurus)

We quantified the specific dynamic action (SDA) resulting from the ingestion of various meal types in Burmese pythons (Python molurus) at 30 degrees C. Each snake was fed a series of experimental meals consisting of amino acid mixtures, simple proteins, simple or complex carbohydrates, or lipids as well as meals of whole animal tissue (chicken breast, beef suet, and mouse). Rates of oxygen consumption were measured for approximately 4 d after feeding, and the increment above standard metabolic rate was determined and compared to energy content of the meals. While food type (protein, carbohydrate, and lipid) had a general influence, SDA was highly dependent on meal composition (i.e., amino acid composition and carbohydrate structure). For chicken breast and simple carbohydrates, the SDA coefficient was approximately one-third the energetic content of the meal. Lard, suet, cellulose, and starch were not digested and did not produce measurable SDA. We conclude that the cost of de novo protein synthesis is an important component of SDA after ingestion of protein meals because (1) simple proteins, such as gelatin and collagen, did not stimulate levels of SDA attained after consumption of complete protein, (2) incomplete mixtures of amino acids failed to elicit the SDA of a complete mixture, and (3) the inhibition of de novo protein synthesis with the drug cycloheximide caused a more than 70% decrease in SDA. Stomach distension and mechanical digestion of intact prey did not cause measurable SDA.     

Specific dynamic action and carbon incorporation in Calanus finmarchicus copepodites and females

Specific dynamic action (SDA) and incorporation of carbon into protein, lipids, and polysaccharides were measured in copepodite CV and adult female Calanus finmarchicus during the spring/summer growth season in Raunefjorden, Norway. Organic carbon from the food (Rhodomonas baltica) was allocated differently in the two developmental stages. Copepodites incorporated 50-80% of the carbon into lipids and only 7-22% into protein. Carbon incorporation into protein was higher in females, constituting 23% in May at 7 °C and 34% in June at 11 °C. This resulted in significant differences in the magnitude of SDA, measured as the increase in oxygen consumption during and after an 8-h feeding event. On average, the rate of carbon incorporation into protein was 2 times higher and the magnitude of SDA was 2.5 times higher in females than in copepodites. There was a significant correlation between the magnitude of SDA and carbon incorporation into protein suggesting that SDA is linked to protein synthesis. When comparing ATP equivalents of the magnitude of SDA with ATP equivalents of the total amount of carbon incorporated, more energy was produced than consumed. This supports speculations of an energy demand associated with a rapid turnover of newly synthesised protein.     

Measles Virus Ribonucleic Acid and Protein Synthesis: Effects of 6-Azauridine and Cycloheximide on Viral Replication

Cycloheximide and 6-azauridine were employed to study the time course of measles virus protein and nucleic acid syntheses in AV3 cells. Synthesis of ribonucleic acid (RNA) essential for infectivity was first detected at 6 hr and increased concurrently with the formation of essential protein. Maximum levels of virus-specific RNA and protein were present by 18 hr, a time when only 5% of progeny virus was detected. Essential RNA and protein syntheses preceded the formation of infectious virus by at least 10 to 12 hr. The time course of RNA and protein syntheses essential for the formation of complement-fixing (CF) antigen and salt-dependent agglutinin (SDA) was also determined. RNA synthesis essential for the formation of SDA was first detected at 2 hr and was present maximally by 6 hr, whereas SDA-protein increased concurrently with the protein essential for infectivity. This suggested that the last protein essential for infectivity may be SDA. RNA synthesis essential for the formation of CF antigen was first detected at 4 hr, while CF-protein increased at 5 hr and preceded SDA-protein and protein essential for infectivity by approximately 3 hr. Reversal of inhibition of protein synthesis by cycloheximide indicated that early protein synthesis (1 to 3 hr) was required for the formation of infectious virus. The data suggest that the relatively long eclipse period observed with measles virus is related to a long maturation period rather than to late formation of early proteins, viral RNA, or structural proteins.     

Thyroid-induced changes in the growth of the Liver,Kidney, and Diaphragm of Neonatal Rats

Eu-, hypo- and hyper-thyroid rats were studied 12 days postpartum. Hypothyroidism was induced by administering propylthiouracil (PTU) via the mother's drinking water beteen late gestation and throughout lactation. This procedure effectively blocked the normal early postnatal surge of T₃ and T₄. In contrast, hyperthyroidism was induced in the young pups by daily injections of T₄ from day 3 postpartum. The effects of these experimental manipulations of thyroid status on the rates of protein turnover and growth of the liver, kidney, and diaphragm were studied and compared with measurements made on appropriate euthyroid control tissues. Tissue rates of protein synthesis were decreased in response to hypothyroidism with consequent growth retardation of all three tissues and the whole animal. In contrast, the three body tissues responded very differently to the induction of hyperthyroidism. Hepatic rates of protein synthesis and growth were completely unaffected by thyroid excess. The response of the diaphragm was essentially the reverse of that seen with hypothyroidism, i.e., the enhanced rates of protein synthesis and protein degradation leading to muscle hypertrophy. The rates of protein turnover in the kidney were also increased, but unlike the diaphragm the net result was renal atrophy. Clearly, thyroid hormones influence the normal rapid growth of the neonate and its individual tissues. However, beyond a certain concentration the threshold of responsiveness to these hormones seems to vary between individual tissues. © 1994 Wiley-Liss, Inc.     

Influence of two different green algal diets on specific dynamic action and incorporation of carbon into biochemical fractions in the copepod Acartia tonsa

Previous studies have shown that the two green algae Tetraselmissp. (Prasinophyceae) and Dunaliella tertiolecta (Chlorophyceae)induce high and low egg production rates in Acartia tonsa. Theprimary goal of the present study was to investigate if thisis attributable to differences in the specific dynamic action(SDA) of the two diets. Secondly, we wanted to investigate ifany qualitative differences in the incorporation of nutritionalconstituents from the two diets are influencing SDA. The functionalresponse of ingestion was very different with the two diets.Ingestion of T. impellucida was relatively high even at lowfood concentrations with a maximum of 19 µg C ind^-1 day^-1.The functional response was more clearly sigmoidal on D. tertiolectawith a maximum of 7.3 µg C ind^-1 day^-1. The higher ingestionrate of T. impellucida also induced higher respiration rates.Maximum respiration rates were 3.0 nl O₂ ind^-1 min^-1 on T. impellucidaand 1.5 nl O₂ ind^-1 min^-1 onD. tertiolecta. This created significantlydifferent SDA coefficients: 0.19 on T. impellucida and 0.06on D. tertiolecta, which implies that the magnitude of SDA isstrongly influenced by the composition of the diet. The incorporationof carbon into lipids was significantly higher on D. tertiolecta.However, because of lack of longer chain fatty acids in D. tertiolectathe copepods did not benefit from this. Thus, the proportionof carbon allocated to egg lipids was much lower than when feedingon T. impellucida. Acartia tonsa incorporated relatively morecarbon into proteins when feeding on T. impellucida than onD. tertiolecta. Since protein synthesis is energetically verydemanding this is probably the reason for the higher SDA coefficientin those feeding on T. impellucida.     

Specific dynamic action in the shore crab,Carcinus maenas (L.), in relation to acclimation temperature and to the onset of the emersion response

The rate of oxygen uptake (MO(2)) of shore crabs following a period of fasting varied directly with acclimation temperature, with a Q(10) of 2.96 between 7 degrees and 15 degrees C and a Q(10) of 2.11 between 15 degrees and 22 degrees C. The factorial rise in MO(2) following a meal (specific dynamic action [SDA]) ranged between 1.9 and 3.1 and varied with temperature, being highest at 15 degrees C and significantly lower at both 7 degrees and 22 degrees C, despite similar ration sizes in all groups. At 7 degrees C, the SDA coefficient and magnitude were significantly lower than at 15 degrees C, possibly due in part to the inhibition of protein synthesis. Both the time to peak and the duration of the SDA response were inversely related to temperature. SDA coefficients were inversely related to the amount of food consumed. The critical oxygen tension of inspired water (P(I)O(2)), which evoked the emersion response in fasted animals, increased with temperature and further increased at each temperature when the animals were fed. Thus, the threshold P(I)O(2) evoking the emersion response is directly related to relative metabolic oxygen demand in Carcinus.     

The relationship between specific dynamic action and growth in grass carp, Ctenophavyngodon idella (Val.)

Individual grass carp, Ctenopharyngodon idellu , were maintained in a respirometer for a month and fed pelleted diets containing various proportions of carbohydrate, fat and protein at different ration levels. Oxygen consumption was measured continuously, allowing the effects of consecutive daily feeding on respiration to be studied. The relationships established between daily food intake and oxygen consumption showed that, on average, 23.3% (high protein diet), 15.3% (high carbohydrate diet), 20.7% (high lipid diet) and 7.0% ( Lemnu diet ) of the absorbed energy was partitioned into specific dynamic action (SDA). (Here the term SDA is used to describe the oxygen consumption of a feeding fish in excess of the routine metabolic rate.) In terms of the overall energy budgets of growing fish, SDA represented between 12 and 58% of the total heat lost over the experimental period and was equivalent to between 14 and 33% of the consumed energy. Ration was positively correlated with heat loss due to total respiration ( r = 0.881) and with heat loss due to SDA ( r = 0.762). As ration increased, the size of SDA relative to total respiration increased. Significant positive correlations were found between oxygen consumption (total or due to SDA) and specific growth rate, and between oxygen consumption and the deposition of protein and energy. However, growth rate had a minimal influence on daily oxygen consumption when compared with food intake.     

Effects of dietary soybean protein levels on metabolic response of the southern catfish, Silurus meridionalis

A closed respirometer was used to measure oxygen consumption of the southern catfish Silurus meridionalis fed with six isonitrogenous (48% crude protein) diets replacing 0%, 13%, 26%, 39%, 52% and 65% fish meal (FM) protein by soybean meal (SBM) protein, in order to investigate the effects of dietary soybean protein level (SPL) (replacing FM) on metabolic rates of the southern catfish. The results showed that there were no significant differences in routine metabolism among dietary treatments. Either the total metabolic rate or specific dynamic action (SDA) was positively correlated with assimilated food energy at each diet, respectively (P<0.05). The SDA coefficient (means the energy spent in metabolism per unit of assimilated dietary energy) significantly increased with increasing dietary SPL (P<0.05). Fish fed the diet with 13% SPL had a significantly lower SDA coefficient (0.1528) than fish fed the diet with 52% or 65% SPL (0.1826 or 0.1932) (P<0.05). However, there were no significant differences in SDA coefficient among fish fed the diets with 13%, 26% and 39% SPL (P>0.05). Results of the present study suggested that an imbalance of essential amino acids at higher dietary SPL resulted in more energy channeled into metabolism, and subsequently increased the SDA coefficient.     

Effects of prey type on specific dynamic action, growth, and mass conversion efficiencies in the horned frog, Ceratophrys cranwelli

To be most energetically profitable, predators should ingest prey with the maximal nutritional benefit while minimizing the cost of processing. Therefore, when determining the quality of prey items, both the cost of processing and nutritional content must be considered. Specific dynamic action (SDA), the increase in metabolic rate associated with feeding in animals, is a significant processing cost that represents the total cost of digestion and assimilation of nutrients from prey. We examined the effects of an invertebrate diet (earthworms) and a vertebrate diet (newborn mice) on mass conversion efficiencies, growth, and SDA in the Chacoan horned frog, Ceratophrys cranwelli. We found the earthworm diet to be significantly lower in lipid, protein, and energy content when compared to the diet of newborn mice. Growth and mass conversion efficiencies were significantly higher in frogs fed newborn mice. However, mean SDA did not differ between frogs fed the two diets, a finding that contradicts many studies that indicate SDA increases with the protein content of the meal. Together, our results indicate that future studies evaluating the effect of meal type on bioenergetics of herpetofauna are warranted and may provide significant insight into the underlying factors driving SDA.     

Specific dynamic action and feeding metabolism in common carp,Cyprinus carpio L.

• 1.1. Oxygen uptake attributable to Specific Dynamic Action (SDA) was measured in common carp, Cyprinus carpio L. (63.6–84.0 g) fed on 20, 35 and 50% dietary protein at 0.40 to 1.00% ration levels at 28°C.
• 2.2. After feeding both SDA magnitude and mean peak oxygen consumption increased directly with dietary protein and ration levels. SDA duration was not significantly related to dietary protein but significantly increased with ration levels.
• 3.3. SDA coefficients were 8.99, 13.51 and 15.94% with 20, 35 and 50% dietary protein showing a direction relationship to the protein content. The SDA coefficient did not change with ration size.
• 4.4. SDA models resulting from this work are of great interest for the aquaculturist, as post-feeding oxygen requirements in an intensive fish culture can be predicted where dietary protein and ration levels are known.

     

Liver protein synthesis. Molecular weight distribution of pulse-labeled polypeptide chains in normal and thyroidectomized rats.

Equations are presented for determination of elongation rate in vivo for a heterogenous population of polypeptide chain molecular weights. The distribution of pulse-labeled polypeptide chains in rat liver deoxycholate-soluble protein has been obtained by sodium dodecyl sulfate-gel electrophoresis and used to compute a theoretical curve for determination of synthesis time of a 50000 mol. wt. polypeptide chain (tc50). Values of tc50 for normal and thyro-parathyroidectomized Long-Evans male rats were 1.2 and 1.75 min, respectively, representing protein synthetic rates of about 7.5 and 5.1 mg protein/g liver/h. No difference in the molecular weight profile of liver polypeptide chains on the basis of labeling or Amido-black staining was observed between the two groups. The distributions of radioactivity before and after secretion of labeled plasma protein are compared. The role of protein-synthetic rate in the changing enzyme levels associated with thyroid hormone is discussed.     

Metabolism and functional effects of plant-derived omega-3 fatty acids in humans

Alpha-linolenic acid (ALA) is an essential fatty acid and the substrate for the synthesis of longer-chain, more unsaturated ω-3 fatty acids, eicosapentaenoic acid (EPA), docosapentaenoic acid and docosahexaenoic acid (DHA). EPA and DHA are associated with human health benefits. The primary source of EPA and DHA is seafood. There is a need for sustainable sources of biologically active ω-3 fatty acids. Certain plants contain high concentrations of ALA and stearidonic acid (SDA). Here we review the literature on the metabolism of ALA and SDA in humans, the impact of increased ALA and SDA consumption on concentrations of EPA and DHA in blood and cell lipid pools, and the extent to which ALA and SDA might have health benefits. Although it is generally considered that humans have limited capacity for conversion of ALA to EPA and DHA, sex differences in conversion to DHA have been identified. If conversion of ALA to EPA and DHA is limited, then ALA may have a smaller health benefit than EPA and DHA. SDA is more readily converted to EPA and appears to offer better potential for health improvement than ALA. However, conversion of both ALA and SDA to DHA is limited in most humans.     

Dose response of protein turnover in rat skeletal muscle to triiodothyronine treatment

Skeletal muscle protein turnover has been examined in thyroidectomized rats treated with 0, 0.3, 0.75, 2, 20 and 100 micrograms triidothyronine/day for 7 days by implanted osmotic minipump. Protein synthesis in gastrocnemius, plantaris and soleus muscle were measured in vivo by the constant infusion method and protein degradation estimated as the difference between gross and net rates of synthesis. Serum levels of triidothyronine (T3) and insulin were also measured in addition to oxygen consumption rates in some cases. Compared with untreated intact rats muscle growth rates were unchanged at 0.3, 0.75 and 2 micrograms T3/day and, judging by plasma T3 levels, 0.75 microgram T3/day was a replacement dose. Slowing of growth was evident in the untreated thyroidectomized rats mid-way through the 7 day experimental period (6-7 days after throidectomy). High doses of T3 (20 and 100 micrograms/day) promptly supressed growth but there was subsequent recovery. Protein synthesis and degradation were generally lower in the hypothyroid state and normal or elevated in the hyperthyroid state. The changes in protein synthesis were mediated by changes in both RNA concentration and RNA activity (protein synthesis per unit RNA). Gastrocnemius and plantaris muscles were most responsive in the hypothyroid range. Since protein synthesis is particularly depressed in these muscles in malnutrition, the fall in protein degradation induced by the lowered thyroid status in this condition will be an important adaptive response to conserve protein. The increased protein turnover in the hyperthyroid rats was most marked in the soleus muscle and it is argued that this is necessary to allow the changes in protein composition and metabolic character which occur in response to hyperthyroidism in this muscle.     

Effects of acclimation and acute temperature change on specific dynamic action and gastric processing in the green shore crab, Carcinus maenas

The effects of temperature acclimation and acute temperature change were investigated in postprandial green shore crabs, Carcinus maenas. Oxygen uptake, gut contractions and transit rates and digestive efficiencies were measured for crabs acclimated to either 10 °C or 20 °C and subsequently exposed to treatment temperatures of 5, 15, or 25 °C. Temperature acclimation resulted in a partial metabolic compensation in unfed crabs, with higher oxygen uptake rates measured for the 10 °C acclimated group exposed to acute test temperatures. The Q₁₀ values were higher than normal, probably because the acute temperature change prevented crabs from fully adjusting to the new temperature. Both the acclimation and treatment temperature altered the characteristics of the specific dynamic action (SDA). The duration of the response was longer for 20 °C acclimated crabs and was inversely related to the treatment temperature. The scope (peak oxygen consumption) was also higher for 20 °C acclimated crabs with a trend towards an inverse relationship with treatment temperature. Since the overall SDA (energy expenditure) is a function of both duration and scope, it was also higher for 20 °C acclimated crabs, with the highest value measured at the treatment temperature of 15 °C. The decline in total SDA after acute exposure to 5 and 25 °C suggests that both cold stress and limitations to oxygen supply at the temperature extremes could be affecting the SDA response. The contractions of the pyloric sac of the foregut region function to propel digesta through the gut, and contraction rates increased with increasing treatment temperature. This translated into faster transit rates with increasing treatment temperatures. Although pyloric sac contractions were higher for 20 °C acclimated crabs, temperature acclimation had no effect on transit rates. This suggests that a threshold level in pyloric sac contraction rates needs to be reached before it manifests itself on transit rates. Although there was a correlation between faster transit times and the shorter duration of the SDA response with increasing treatment temperature, transit rates do not make a good proxy for calculating the SDA characteristics. The digestive efficiency showed a trend towards a decreasing efficiency with increasing treatment temperature; the slower transit rates at the lower treatment temperatures allowing for more efficient nutrient absorption. Even though metabolic rates of 10 °C acclimated crabs were higher, there was no effect of acclimation temperature on digestive efficiency. This probably occurred because intracellular enzymes and digestive enzymes are modulated through different control pathways. These results give an insight into the metabolic and digestive physiology of Carcinus maenas as it makes feeding excursions between the subtidal and intertidal zones.