Early development of milkfish: effects of salinity on embryonic and larval metabolism, yolk absorption and growth

During embryogenesis of Chanos chanos , more than half of the yolk was consumed and the majority of it was converted into larval tissue. Salinity affected both yolk absorption and embryonic and larval growth. Larvae hatched in 20% had larger yolk reserves but were smaller and grew more slowly than larvae in 35 and 50%. Larvae hatched in 35 and 50% had equal amounts of yolk but those from 35% were larger. Oxygen consumption rates increased during development (from 0.06 ± 0.01 μl O₂ egg^–1 h^–1 by blastulae to 0.37 ± 0-01 μl O₂ egg^–1 h^–1 by prehatch embryos and 0–43 ± 0–03 μl O₂ larva ^–1 h ^–1 by newly-hatched larvae) and were significantly affected by salinity. Eggs and yolk-sac larvae incubated in 35% consumed more oxygen than those in the low and high salinities. Salinity affected both the rate and pattern of yolk utilization but salinity-related differences in metabolism, yolk absorption, and growth were not related directly to the osmotic gradient. Low salinity retarded yolk absorption while high salinity reduced yolk utilization efficiencies. Differences in oxygen consumption rates were probably related to variations in the relative amounts of metabolically active embryonic and larval tissue and/or higher activity levels rather than differential osmoregulatory costs. 35% is probably the most suitable salinity for incubation and larval rearing of milkfish.     

Oxygen uptake in developing eggs and larvae of the cod, Gadus morhua L.

Unfertilised cod eggs showed a mean oxygen uptake rate at 5°C of 0.089 μl O₂, dry wt.⁻¹ h⁻¹; this gradually rose to 0.768 μl O₂ mg dry wt.⁻¹ h⁻¹ in eggs about to hatch. From hatching to complete yolk absorption larvae respired at 1.6 μl O₂, mg dry wt.⁻¹ h⁻¹. During starvation following yolk absorption, uptake fell significantly to 1.1 μl O₂, mg dry ⁻¹ h⁻¹. Much of this decrease in oxygen consumption was shown to be caused by reduction in activity. Loss of weight during the embryo and larval phases could not easily be reconciled with total oxygen consumption; it is suggested that cod embryos and larvae may not rely solely upon endogenous energy reserves during development.     

Oxygen consumption by eggs and larvae of striped mullet, Mugil cephalus, in relation to development, salinity and temperature

Oxygen consumption rates during embryonic and the first 38 days of larval development of the striped mullet were measured at 24° C by differential respirometry. Measurements were obtained at the blastula, gastrula and four embryonic stages, and at the yolk-sac, preflexion, flexion and post-flexion larval stages.
Oxygen uptake rates of eggs increased linearly from 0.024 μl O₂ per egg h^-1 (0·323 μl O₂ mg^-1 dry wt h^-1) by blastulae to 0·177 μlO₂ per egg h^-1 (2·516 μlO₂mg ¹dry wth^-1) by embryos prior to hatching. Respiration rates did not vary significantly among four salinities (20,25, 30, 35%₀).
Larval oxygen consumption increased in a curvilinear manner from 0·243 μl O₂ per larva h^-1 shortly after hatching to 18·880 μl O₂ per larva h^-1 on day 38. Oxygen consumption varied in direct proportion to dry weight. Mass-specific oxygen consumption rates of preflexion, flexion, and postflexion larvae did not change with age (10·838 μl O₂ mg ¹dry wt h^-1).
Larval oxygen consumption rates did not vary significantly among salinities 10–35%. Acute temperature increases elicited significant increases in oxygen consumption, these being relatively greater in yolk-sac larvae ( Q₁₀ = 2·75) than in postflexion larvae ( Q₁₀ = 1·40).     

Effects of development, temperature and salinity on metabolism in eggs and yolk-sac larvae of milkfish, Chanos chanos (Forsskål)

Oxygen uptake rates and yolk-inclusive dry weiGhts were measured during the egg and yolk-sac larval stages of milkfish, Chanos chanos (Forsskal). Oxygen uptake by eggs and yolk-sac larvae was measured to assess the effects of four salinities (20,25,30,35 ppt) at 28°C. The effects of three temperatures (23,28,33°C) on oxygen uptake by yolk-sac larvae were determined at a salinity of 35 ppt. Dry weights were measured throughout embryonic development at 28°C and the yolk-sac stage at 23.28 and 33°C.
Oxygen uptake rates of eggs increased more than fivefold during embryogenesis (0.07±0.03 to 0.40 ± 03 μl O₂ egg ⁻¹ h ⁻¹;blastula to prehatch stage). Larval oxygen uptake did not change with age but was affected by rearing temperature (0.33 ± 0.08, 0.44 ± 0.07 and 0.63 ± 0.13 μl O₂ larva ⁻¹ h⁻¹ at 23, 28 and 33°C, respectively; Q₁₀= 1.93). Acute temperature changes from 28 to 33°C caused significant increases in oxygen uptake by embryos (Q ₁₀= 1.69–3.58) and yolk-sac larvae (Q ₁₀=2.55). Salinity did not affect metabolic rates.
Dry weight of eggs incubated at 28°C decreased 13% from fertilization to hatching. Incubation temperatures from 23–33°C did not affect dry weights at hatching. Rearing temperatures significantly affected the rate of larval yolk absorption (Q ₁₀= 2.25).     

Effect of body size, temperature, and salinity on the routine metabolism of larval and juvenile spotted seatrout

Routine oxygen consumption rates of young spotted seatrout Cynoscion nebulosus (Sciaenidae) were measured over a range of temperatures (24, 28, 30 and 32° C) and salinities (5, 10, 20, 35 and 45). Larvae and juveniles, 4·1–39·5 mm standard length ( L _S), ranging several orders of magnitude in dry body mass were used to estimate the mass–metabolism relationship. Oxygen consumption (μl O₂ larva⁻¹ h⁻¹) scaled isometrically with body mass for larvae <5·8 mm L _S(phase I, slope = 1·04) and allometrically thereafter (phase II, slope = 0·78). The inflection in the mass–metabolism relationship coincided with the formation of the hypural plate and an increase in the relative tail size of larvae. Salinity did not have a significant effect on routine metabolism during phase I. Temperature and salinity significantly affected routine metabolism during phase II of the mass–metabolism relationship. The effect of salinity was temperature dependent, and was significant only at 30° C. Response surfaces describing the environmental influences on routine metabolism were developed to provide a bioenergetic basis for modelling environmental constraints on growth.     

Seasonal differences in routine oxygen consumption rates of the bonnethead shark

Routine oxygen consumption rates of bonnethead sharks, Sphyrna tiburo , increased from 141·3±29·7 mg O₂ kg⁻¹ h⁻¹ during autumn to 218·6±64·2 mg O₂ kg⁻¹ h⁻¹ during spring, and 329·7±38·3 mg O₂ kg⁻¹ h⁻¹ during summer. The rate of routine oxygen consumption increased over the entire seasonal temperature range (20–30° C) at a Q ₁₀=2·34.     

Routine metabolism of juvenile spot, Leiostomus xanthums (Lacépède), as a function of temperature, salinity and weight

Routine oxygen consumption rates of juvenile spot, Leiostomus xanthums , were measured over a range of temperatures, salinities and fish weights. As predicted, Q O₂ increased with temperature and decreased with body weight. However, Q O₂ decreased with decreasing salinity and did not show the expected minimum at isosmotic concentrations. The data are best described by the relationship: log₁₀ Q O₂ (mg O₂ g⁻¹ h⁻¹) = 0.129 log_lo salinity (%0) + 1.604 log₁₀ temperature (°C)-0.1401og₁₀(g)-2.767.     

The ontogeny of drinking in the rainbow trout, Oncorhynchus mykiss (Walbaum)

Drinking was measured by the rate of uptake of tritiated dextran in fresh water by the alevins and fry of rainbow trout at various stages of development during a 40-day post-hatch period. Drinking increased almost S-fold during the initial 16 day yolk-sac stage. Drinking rate increased most between 16 and 23 days, the transitional period between yolk-sac absorption and first feeding. The maximum weight-specific drinkingrateof 3.24μlg⁻¹ h⁻¹ recorded for 40-day-old fry was higher than previously recorded for adults. Abrupt transfer from an adaptation temperature of 10 to 19° C increased drinking significantly (Q₁₀= 1.2), but sudden transfer to 1 1‰ salinity sea water caused a substantial fall in drinking rate in 23-day fry. A 24-h period of adaptation to 1 1‰ and 15‰ salinity restored drinking to a rate similar to that in fresh water. The water drunk by 31-day fry fed to satiation was initially higher than by unfed fry, but drinking rates subsequently fell below the control level. The results are discussed in terms of osmoregulation and the uptake of dissolved or suspended substances by the intestinal tract for the purpose of immunization.     

On the diel rhythms in metabolism and activity of post-hatching lesser spotted dogfish, Scyliorhinus canicula

The diel rhythms in metabolic rate ( M_R ) and activity level ( A_L ) were measured for single post-hatching dogfish (weight range, 2.76–10.61 g) at 15° C by the indirect calorimetric method of rate of oxygen consumption ( V O₂) and by video-observation respectively, over a period of 72 b. The mean VO ₂ increased from 62.0 (s.e. 2.9) mg O₂ kg⁻¹ h⁻¹ in the daylight hours to 85.5 (s.e. 3.1) mg O₂ kg⁻¹ h⁻¹ during the dark (light regíme, 12 h L: 12 h D). The simultaneous measurement of A _L also showed mean night elevation from 0.6 (s.e. 0.2) min h⁻¹ in the light phase to 14.5 (s.e. 1.6) min h⁻¹ during the darkness. Bimodal nocturnal activity (BNA) was exhibited by the post-hatching dogfish within the 12 h dark period, with V O₂ increasing from 71.4 (s.e. 2.8) mg O₂ kg⁻¹ h⁻¹ before 01.00 hours to 99.5 (s.e. 4.2) mg O₂ kg⁻¹ h⁻¹ after 01.00 hours. Similarly, A _L also increased from 8.9 (s.e. I.7)min h⁻¹ before 01.00 hours to 21.1 (s.e. 2.8) min h⁻¹ after 01.00 hours. The importance of the results presented to the natural behavioural ecology of the hatching dogfish are discussed.     

Effect of inhibitors on ammonium assimilation in Chlorella sorokiniana in light and darkness

In N-sufficient cells of Chlorella sorokiniana Shihira and Krauss strain 211/8K (CCAP of Cambridge University), assimilation of ammonium was strictly dependent on light and CO₂, and was severely inhibited by 100 μ M atrazine or 10 μ M 3-(3,4-dichlorophenyl)-1, l-dimethylurea (DCMU). In N-limited cells, assimilation of NH₄⁺ took place at similar rates in both light and darkness, which were 1.6-fold higher than the rate of light-dependent assimilation by N-sufficient cells. Assimilation by N-limited cells was inhibited by l -methionine- dl -sulfoximine (MSX), but not by atrazine or DCMU.
The rate of photosynthetic O₂ evolution was 2.9±0.9 mmol ml⁻¹ packed cell volume (PCV) h⁻¹ in N-sufficient cells, and 0.64±0.12 mmol ml⁻¹ PCV h⁻¹ in N-limited cells. In the latter resupply of ammonium resulted in a rapid activation by 22%;, followed by a time-dependent increase of the photosynthetic O₂ evolution, which after 12 h reached the same rate as in N-sufficient cells.
Respiratory consumption of oxygen in darkness in N-sufficient and N-limited cells was 0.10±0.03 and 0.11±0.02 mmol ml⁻¹ PCV h⁻¹, respectively. Addition of ammonium was without effect on respiration of N-sufficient cells, but resulted in a 4-fold stimulation of respiration of N-limited cells. Such stimulation took place also in cells treated with DCMU, atrazine, or MSX, and it was also promoted by methylammonium. The stimulation of respiration lasted for several hours.     

Scaling of oxygen consumption of Lake Magadi tilapia, a fish living at 37°C

Rates of oxygen consumption were measured in the geothermal, hot spring fish, Oreochromis alcalicus grahami by stopped flow respirometry. At 37° C, routine oxygen consumption followed the allometric relationship: V o₂=0.738 M ^0.75, where V o₂ is ml O₂ h ⁻¹ and M is body mass (g). This represents a routine metabolic rate for a 10 g fish at 37° C of 0.415 ml O₂ g⁻¹ h ⁻¹ (16.4 μmol O₂ g ⁻¹ h ⁻¹). Acutely increasing the temperature from 37 to 42° C significantly elevated the rate of O₂ consumption from 0.739 to 0.970 ml O₂ g ⁻¹ h ⁻¹ ( Q ₁₀=l.72). In the field, O. a. grahami was observed to be 'gulping' air from the surface of the water especially in hot springs that exceeded 40° C. O. a. grahami may utilize aerial respiration when O₂ requirements are high.     

Individual variation in the rate of oxygen consumption by zebrafish embryos

A sensitive microsensor‐based method was used to measure oxygen consumption of individual zebrafish Danio rerio embryos at 6 h intervals from 24 to 75 h post‐fertilization. An increase in oxygen consumption rates from 4·54 to 8·29 nmol O₂ h⁻¹ was found during this period. At the individual level the differences in oxygen consumption rates caused the total oxygen consumption from 24 to 75 h post‐fertilization to vary between 0·261 and 0·462 μmol O₂ per individual with a mean of 0·379 μmol O₂ per individual. A separate carbon mass balance study corroborated the mean total oxygen consumption obtained by yielding a respiratory quotient of 0·80 for this period. These results suggest that there is significant intraspecific variation in the metabolic rate of developing zebrafish embryos, which may influence other early life‐history traits such as growth and starvation resistance.     

Metabolic utilization of body stores during the early life of whitefish, Coregonus lavaretus L.

Patterns of oxygen consumption, ammonia and urea excretion were monitored during late embryogenesis, i.e. 5 days before mass hatching and 12 days during the free-swimming stage of whitefish larvae, Coregonus lavaretus. Oxygen consumption increased from 1.31 to 2.53 mgO₂ h⁻¹× 10³ eggs⁻¹ at hatching. Fasted, free-swimming larvae showed increasing oxygen consumption to the tenth day after hatching when it reached 5.52 mgO₂h⁻¹× 10³ larvae⁻¹. Ammonia and urea excretion increased during pre-hatching period from 52.1 to 163.2 and 26.8 to 51.4 μgh⁻¹× 10³ eggs⁻¹, respectively. The nitrogen excretion rate increased between the sixth and tenth day of fasting, i.e. for ammonia from 117.7 to 160.9 and for urea from 35.8 to 52.5 μg h⁻¹× 10³ larvae⁻¹. Cumulative data on nitrogen and energy metabolism indicated that during late embryogenesis, and up to the fifth day after hatching, protein dominated in the energy expenditure. During the free swimming stage, the ratio of fat to protein in energy expenditure rose from 0.86 to 1.99. Combined data for several fish species indicated high dependance of oxygen uptake during the hatching period on egg size and temperature.     

A method for long-term measurement of respiration in intertidal fishes during simulated intertidal conditions

Aquatic and aerial respiration of the amphibious fishes Lipophrys pholis and Periophthalmus barbarus were examined using a newly designed flow-through respirometer system. The system allowed long-term measurements of oxygen consumption and carbon dioxide release during periods of aquatic and aerial respiration. The M o ₂ of L. pholis , measured at 15° C, was 2·1 μmol O₂ g^–1 h^–1 during aquatic and 1·99 μmol O₂ g^–1 h^–1 during aerial exposure. The corresponding values of the M co₂ were 1.67 and 1.59 μmol O₂ g^–1 h^–1 respectively, giving an aquatic respiratory exchange ratio (RER) of 0·80 and an aerial RER of 0·79. The M o₂ of P. barbarus , measured at 28°C, was 4·05 μmol O₂ g^–1 h^–1 during aquatic and 3·44 μmol O₂ g^–1 h^–1 during aerial exposure. The corresponding values of the Mco₂ were 3·29 μmol CO₂ g^–1 h^–1 and 2·63 μmol CO₂ g^–1 h^–1 respectively, giving an aquatic RER of 0·81 and an aerial RER of 0·77. While exposed to air for at least 10 h, both species showed no decrease in metabolic rate or carbon dioxide release. The RER of these fishes equalled their respiratory quotient. After re-immersion an increased oxygen consumption, due to the payment of an oxygen debt, could not be detected.     

Oxygen consumption in Caridina nilotica (Decapoda: Atyidae) in relation to temperature and size

SUMMARY. The oxygen consumption of shrimps ranging from 1 to 30 mg dry mass was determined at 18, 24 and 30°C using a continuous flow recording respirometer based upon a Clark-type oxygen electrode. Respiration (ascribed to routine metabolism) is described by the power curve: R = a M^b , ( R =μg O₂ h⁻¹, M = mg dry mass), which gives values of a = 1.632, 2.564 and 4.181, and b = 0.800, 0.898, and 0.793, at 18, 24 and 30°C respectively. The single expression, R = 0.008 T ^1.829 M ^0.830 provides a reasonable prediction of respiration as a combined function of shrimp size ( M ) and temperature (T, °C). Using an energy equivalent of 14.14 J mg O₂⁻¹ estimates of the energy requirements ( E , J h⁻¹ 10⁻³) of routine metabolism are given by the expression: E = 0.115 T ^1.829 M ^0.830.
Variability in oxygen consumption values between individuals is discussed and the observations on C. nilotica are compared with other crustacean studies.     

Effects of rapid changes in oxygen concentration on the respiration of carrot roots

Rates of CO₂ production and O₂ consumption from aged disks of carrot ( Daucus carota L.) root tissues were measured for 4 h after they were transferred from 21% to 0, 1, 2, 4 or 8% O₂ in gas mixtures. A transient peak in the rate of CO₂ production started 5 to 7 min after transfer to 2% or lower O₂ mixtures and peaked at 50 min. After the peaks in CO₂ production from the 0, 1 and 2% O₂ treatments and after the stable production from the 4 and 8% O₂ treatments, the rate of CO₂ production from all low O₂ treatments started to decline at 50 min, reaching stable rates by 160 to 240 min. Concentrations of lactate and ethanol that were significantly higher than the 21% O₂ controls had started to accumulate in disks between 10 and 50 min after exposure to atmospheres containing 2% or less O₂. Production of CO₂ started to increase 5 to 7 min after transfer to 0, 1 and 2% O₂, while the initial decline and then rise in pH and the accumulation of ethanol did not occur until 30 min after the change in atmosphere. Ethanol accumulation paralleled the increase in pH; first at 0.4 μmol g⁻¹ h⁻¹ from 30 to 60 min as the pH shifted from 5.97 to 6.11, and then at 0.08 μmol g⁻¹ h⁻¹ from 60 to 100 min as the pH stablized around 6.12. The peak at 50 min in CO₂ production roughly coincided with the shift from the rapid to the slow change in pH and ethanol accumulation.     

Energy metabolism in eggs and larvae of the Senegal sole

Oxygen consumption in Solea senegalensis increased during the egg stage reaching values close to 4 nmol O₂ ind⁻¹ at hatching. After hatching, larval oxygen consumption continued to increase, reaching a maximum rate of 9.97⁻¹±87 nmol O₂ ind⁻¹ h⁻¹ 2 days after the opening of the mouth. Body nitrogen content decreased mainly after exhaustion of yolk reserves. Carbon content decreased during the whole endogenous feeding phase, although it decreased twice as quickly after yolk-sac absorption. The free amino acid (FAA) depletion rate was higher during egg development and the yolk-sac period. Complete yolk absorption coincided with the consumption of the 90% of initial FAA content in the eggs and the remaining FAA were consumed at a lower rate. Based on stoichiometrical calculations, FAA appears to be the most important energy substrate during the egg stage (86%) in the Senegal sole. During the period from hatching to the mouth opening, contributions of FAA and lipids as metabolic fuels were similar (41 and 47%, respectively). The decrease in larval protein content during starvation indicates that amino acids from body protein are used as energy substrates under food deprivation.     

Preparation of Arabidopsis mesophyll protoplasts with high rates of photosynthesis

A simple and quick method is described for rapid isolation of metabolically active mesophyll protoplasts from leaves of Arabidopsis thaliana . The optimal composition of the digestion medium, period of digestion and stability of protoplast preparation were examined. A large number of protoplasts could be prepared within an hour. The isolated protoplasts were intact, stable and metabolically very active, as indicated by their high rates of photosynthetic oxygen evolution. The important factors during the preparation of protoplasts are short time of digestion, composition of medium, use of nylon nets for filtration, centrifugation at low speed and use of pH 7.0 for storage. The highest rate of photosynthesis obtained in these experiments was 130 ± 4 μmol O₂ evolved mg⁻¹ Chl h⁻¹, at 1 m M sodium bicarbonate and at a light intensity of 600 μE m⁻² s⁻¹. The present technique of isolation can be very useful for making Arabidopsis protoplasts for studies on not only metabolic processes, such as photosynthesis, but also metabolomics, proteomics and genomics.     

Simultaneous phototrophic and chemotrophic growth in the purple sulfur bacterium Thiocapsa roseopersicina M1

Abstract The anoxygenic phototrophic purple sulfur bacterium Thiocapsa roseopersicina was grown in illuminated continuous cultures with thiosulfate as growth limiting substrate. Aeration resulted in completely colorless cells growing chemotrophically, whereafter the conditions were changed to a 23 h oxic/1 h anoxic regime. After 11 volume changes at a dilution rate of 0.031 h⁻¹ (35% of μ_max) a time dependent equilibrium was established. During the 23 h oxic periods bacteriochlorophyll a synthesis (BChl a ) was not observed, whereas during the 1 h anoxic periods synthesis was maximal (i.e. 1.1 μg (mg protein)⁻¹ h⁻¹). As a result the BChl a concentration gradually increased from zero to an average value over 24 h of 1.9 μg (mg protein)⁻¹. Concomitantly, the protein concentration increased from 13.9 mg 1⁻¹ during continuous oxic conditions to 28.8 mg 1⁻¹. For comparison, the protein concentration during fully phototrophic growth at an identical thiosulfate concentration in the inflowing medium was 53.7 mg 1⁻¹. The specific respiration rate was 8 μmol O₂ (mg protein)⁻¹ h⁻¹ during full chemotrophic growth and gradually decreased to 3.5 μmol O₂ (mg protein)⁻¹ h⁻¹ after 11 volume changes at the regime employed. These data show that T. rosepersicina is able to simultaneously utilize light and aerobic respiration of thiosulfate as sources of energy. The ecological relevance of the data is discussed.     

Effects of temperature, hypoxia and activity on the metabolism of juvenile Atlantic cod

Standard metabolic rate (SMR), active metabolic rate (AMR) and critical oxygen saturation ( S_crit ) were measured in Atlantic cod Gadus morhua at 5, 10 and 15° C. The SMR was 35.5, 57.0 and 78.2 mg O₂ kg⁻¹ h⁻¹ and S_crit was 16.5, 23.2 and 30.3%, at 5, 10 and 15° C, respectively. Previously reported SMR for Atlantic cod from arctic waters at 4° C was twice that measured at 5° C in the present study. A possible intraspecific latitudinal difference in the SMR is discussed. The AMR was 146.6, 197.9 and 200.4 mg O₂ kg⁻¹ h⁻¹ and the critical swimming speed ( U_crit ) was 1 6, 1.7 and 1.9 at 5, 10 and 15° C, respectively. The maximum oxygen consumption was found to be associated with exercise, rather than recovery from exercise as previously reported in another Study of Cod metabolism.