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.     

Opercular differential pressure as a predictor of metabolic oxygen demand in the starry flounder

The feasibility of using a differential pressure sensor connected to an acoustic telemetry device to monitor opercular activity as a correlate of oxygen consumption was investigated. Four starry flounders Platichthys stellatus were fitted with a miniature differential pressure sensor mounted close to the operculum. A cannula was connected to the sensor and inserted under the operculum, inside the branchial cavity. Measurements of oxygen consumption and opercular activity were carried out over a broad range of metabolic activity, from the post‐surgery stress (high metabolic rate) to routine metabolic rate the following day. Relationships between differential pressure changes (rate and amplitude) were highly correlated with oxygen consumption ( r ² = 0·74 and 0·60 respectively). The results indicate that monitoring opercular activity offers an alternative method for measuring aerobic metabolism in free‐swimming fishes in nature.     

Changes in growth performance and proximate composition in Japanese flounder during metamorphosis

Morphological changes are described in Japanese flounder Paralichthys olivaceus larvae and juveniles with emphasis on growth during the period of metamorphosis. Ontogenetic changes in condition factor and lipid, glycogen and protein levels were also analysed to determine the utilization of stored energy. Fish grew from 6·6 to 20·3 mm L _T(1·15 to 84·4 mg in mass, M ) during the period from 11 to 40 days after hatching (DAH) at 19·3° C. Per cent specific growth rate per day (% G day⁻¹) for wet mass was lowest during the metamorphic phase (21–30 DAH) compared to pre‐(11–20 DAH) and post‐metamorphic (31–40 DAH) phases. When L _T and M were expressed as     , the b value was highest during the pre‐metamorphic phase and lowest during the metamorphic phase. These findings indicate that the developmental changes that occur during metamorphosis of Japanese flounder are closely related to the growth pattern. Moisture, lipid and glycogen contents were also at the lowest level during metamorphosis compared to pre‐ and post‐metamorphosis, which suggest that Japanese flounder use their energy reserves to accomplish metamorphosis due to an apparent decline in feeding during this period.     

Direct measurements of metabolism, activity and feeding behaviour of pike, Esox Zucius L., in the wild, by the use of heart rate telemetry

Heart rate telemetry records of up to 5 days duration were obtained from pike living in Lochs Kinord and Davan, Scotland. Applying metabolic rate correlations it was found that mean metabolic rate (R) was 1.5 times standard metabolic rate (R_a), The fish rarely worked near their metabolic limits. Activity metabolism (R_a) was much higher than estimates based on mean swimming speed and comprised up to 10% of R. Most activity metabolism was the result of localized bursts of activity. Less than 10% of activity showed evidence of oxygen debt. Specific Dynamic Action or feeding metabolism (R_f) comprised 15–25% of R. Food intake estimated from heart rate was 1.5% wet body weight day⁻¹, consumed in the form of small items captured during the day and digested during the afternoon and night.     

Geographical differences in marine feeding of Atlantic salmon post‐smolts in Norwegian fjords

Stomach content analyses were conducted on Atlantic salmon Salmo salar post‐smolt (average size, 119–154 mm fork length, L _F) caught in eight large Norwegian fjord systems along a north–south geographical axis during 1998–2001. In general, post‐smolts from southern Norway showed low feeding intensity in the fjords, whereas extensive feeding was observed in fjords in the northern and middle parts of Norway. The marine diet mainly included different crustaceans and in particular marine pelagic fish larvae (sand-eels Ammodytes spp., herring Clupea harengus and gadoids), but with a substantial spatial and annual variation in prey diversity and feeding intensity. Insects were most frequently taken in the estuary, although fishes often made a large contribution in mass. In contrast, fishes, and to some extent various crustaceans (particularly Hyperiidae, Gammaridae, Euphausiacea and Copepoda) dominated the diet in the middle and outer parts of the fjords, where post‐smolts also fed more extensively than in the inner part. The results indicate that extensive feeding immediately after sea entrance may be more common for post‐smolts in the northern and middle parts of Norway, than in the southern fjords. The observed differences in post‐smolt feeding may be due to spatial and temporal differences in prey availability within and between the different types of fjord systems, and this might influence post‐smolt growth and survival.     

The calpastatin/calpain system in trout Salmo trutta trutta muscle

Many recent reports suggest that the calpastatin/calpain system plays a role in cellular growth and differentiation. Defects of the calpastatin/calpain system have been linked to cellular dysfunctions, apoptosis, myocardial infarct, and dystrophies. The calpastatin/calpain system has also been implicated in post‐mortem tenderization of skeletal muscle through degradation of key myofibrillar and associated proteins, a process of key importance to meat quality. In the present study we investigate the presence and activity of the calpastatin/calpain system in trout muscle samples, collected at 0, 3, 18 and 28 h post‐mortem, by immunohistochemistry method. Calpastatin is a specific endogenous enzyme of cytosol, modulating the ubiquitous calpains. Calpastatin was found in samples obtained in vivo and immediately post‐mortem, but its concentration declined rapidly in samples obtained 3, 18 and 28 h post‐mortem. The ubiquitous m e m‐calpains, which are localized on Z line proteins and activated by intracellular Ca²⁺ increase, showed a rapid decline within 3 h post‐mortem. By contrast p94 calpain, which is specific to skeletal muscle, showed a slow decrease post‐mortem which was independent of intracellular Ca²⁺ increase. Our results suggest that the mechanism of activation and activity of the calpastatin/calpain system in trout is similar to that described in mammals.     

Effects of temperature, body size and feeding on rates of metabolism in young‐of‐the‐year haddock

The mean rate of oxygen consumption (routine respiration rate, R _R, mg O₂ fish⁻¹ h⁻¹), measured for individual or small groups of haddock Melanogrammus aeglefinus (3–12 cm standard length, L _S) maintained for 5 days within flow‐through respiratory chambers at four different temperatures, increased with increasing dry mass ( M _D). The relationship between R _R and M _D was allometric ( R _R = α  M ^b ) with b values of 0·631, 0·606, 0·655 and 0·650 at 5·0, 8·0, 12·0 and 15·0° C, respectively. The effect of temperature ( T ) and M _D on mean R _R was described by     indicating a Q ₁₀ of 2·27 between 5 and 15° C. Juvenile haddock routine metabolic scope, calculated as the ratio of the mean of highest and lowest deciles of R _R measured in each chamber, significantly decreased with temperature such that the routine scope at 15° C was half that at 5° C. The cost of feeding ( R _SDA) was c . 3% of consumed food energy, a value half that found for larger gadoid juveniles and adults.     

Temperature responses are a window to the physiology of dark respiration: differences between CO2 release and O2 reduction shed light on energy conservation

We showed that temperature responses of dark respiration for foliage of Pinus radiata could be approximated by Arrhenius kinetics, whereby E ₀ determines shape of the exponential response and denotes overall activation energy of respiratory metabolism. Reproducible and predictable deviation from strict Arrhenius kinetics depended on foliage age, and differed between R _CO2 and R _O2. Inhibition of oxygen reduction ( R _O2) by cyanide (inhibiting COX) or SHAM (inhibiting AOX) resulted in reproducible changes of the temperature sensitivity for R _O2, but did not affect R _CO2. Enthalpic growth – preservation of electrons in anabolic products – could be approximated with knowledge of four variables: activation energies ( E ₀) for both R _CO2 and R _O2, and basal rates of respiration at a low reference temperature ( R _REF). Rates of enthalpic growth by P. radiata needles were large in spring due to differences between R _REF of oxidative decarboxylation and that of oxygen reduction, while overall activation energies for the two processes were similar. Later during needle development, enthalpic growth was dependent on differences between E ₀ for R _CO2 as compared with R _O2, and increased E ₀( R _O2) indicated greater contributions of cytochrome oxidase to accompany the switch from carbohydrate sink to source. Temperature-dependent increments in stored energy can be calculated as the difference between R _CO2▵ H _CO2 and R _O2▵ H _O2.     

Biochemical, histological and behavioural aspects of visual function during early development of rainbow trout

Retinal structure and concentration of retinoids involved in phototransduction changed during early development of rainbow trout Oncorhynchus mykiss , correlating with improvements in visual function. A test chamber was used to evaluate the presence of optokinetic or optomotor responses and to assess the functionality of the integrated cellular, physiological and biochemical components of the visual system. The results indicated that in rainbow trout optomotor responses start at 10 days post‐hatch, and demonstrated for the first time that increases in acuity, sensitivity to low light as well as in motion detection abilities occur from this stage until exogenous feeding starts. The structure of retinal cells such as cone ellipsoids increased in length as photopic visual acuity improved, and rod densities increased concurrently with improvements in scotopic thresholds (2·2 log₁₀ units). An increase in the concentrations of the chromophore all‐trans‐retinal correlated with improvements of all behavioural measures of visual function during the same developmental phase.     

Inter‐individual differences in rates of routine energy loss and growth in young‐of‐the‐year juvenile Atlantic cod

Inter‐individual differences in rates of routine (non‐feeding) metabolism and growth were evaluated in young‐of‐the‐year (YOY) juvenile Atlantic cod Gadus morhua . Rates of O₂ consumption, CO₂ production and ammonia (TAN) excretion were measured in 64, 25–43 mm standard length ( L _S) YOY growing at different rates (0·27–0·47 mm day⁻¹) in a common rearing tank. Parameter rates ( y ) increased allometrically ( y = a·M^b ) with increasing body mass ( M ) with b ‐values for O₂ production, CO₂ consumption and TAN excretion equal to 0·81, 0·89 and 0·56, respectively. In some cases, residuals from these regressions were significantly negatively correlated to fish growth rate. In no cases did residuals of parameter rates increase with increasing growth rate. These data suggest that, during unfed periods, relatively fast‐growing fish were more metabolically efficient than slower‐growing fish from the same cohort. The fish condition factor, derived from     , also significantly decreased with increasing growth rate. Results indicated differences in both the rates of routine energy loss and the patterns of growth allocation among YOY Atlantic cod. Since these physiological attributes were positively correlated with growth rate, they may be indicative of 'survivors' in field populations.     

Reproductive biology of the milk shark Rhizoprionodon acutus and the bigeye houndshark Iago omanensis in the coastal waters of Oman

Female Rhizoprionodon acutus were found to mature between 62 and 74 cm total length ( L _T) whereas males matured between 63 and 71 cm L _T. The L _T at 50% maturity was 64·3 cm for females and 64·7 cm for males. Litter size varied from one to six embryos, and was positively correlated with maternal L _T. Female embryos outnumbered males by a ratio of 2·3:1. The size at birth was c . 37 cm L _T. Full‐term embryos and post‐partum females were observed during all seasons although their occurrence was highest in spring. Spermatozoa were rarely recorded in the oviducal gland, indicating that this species does not store sperm. It was not possible to generate maturity curves for Iago omanensis but it was evident that females matured by the time they reached 35 cm and males were mature by 31 cm L _T. This species displayed a clearly defined reproductive cycle with parturition occurring primarily in spring, after a gestation period of c . 1 year. Maximal embryo size was 19 cm L _T while maximal litter size was 24 embryos. The oviducal gland appeared to act as a seminal receptacle and it appeared that females may utilize these stores by not mating every year.     

Modelling temperature effects on growth–respiration relations of maize

The temperature dependence of plant growth rate is related to the temperature dependence of respiratory metabolism. To determine how the effects of temperature on respiration rate and efficiency are transmitted to growth, this study measured the dark metabolic heat rate ( q ) and CO₂ production rate ( R _CO2) in excised shoots of seedlings of 14 maize cultivars ( Zea mays L.) at several temperatures. The temperature coefficients of q and R _CO2 differ within a given cultivar and also differ among the cultivars. Both q and R _CO2 exhibit an isokinetic temperature of 20 ± 3 °C. The measured temperature dependences of q and R _CO2 were used to model the temperature dependences of both growth and substrate carbon conversion efficiency. This procedure may be useful in determining the suitability of cultivars for growth in a given climate and in understanding metabolic adaptation to climate.     

Effect of body size on the standard metabolism of horse mackerel

The routine metabolic rate R _R and standard metabolic rate R _S were measured in horse mackerel Trachurus trachurus at 13°C over weight range of 1·4–390 g. A data extraction method rather than the more commonly used method of extrapolating the swimming speed-metabolic rate curves back to zero swimming speed was developed to measure the R _S. The relation between R _R and R _S and weight was expressed as a linear regression with the log transformed data. The mean slope of the regression was 0·752 for R _S and 0·725 for R _S.     

Patterns of oocyte growth, vitellogenin and gonadal steroid concentrations in greenback flounder

The pattern of oocyte development in association with changes of plasma concentrations of vitellogenin (Vtg), 17β‐oestradiol (E₂) and testosterone (T) was investigated in maturing female greenback flounder Rhombosolea tapirina over the first part of a reproductive season (February to June). Examination of oocyte size‐frequency distributions showed that the oocyte developmental pattern in R. tapirina is multiple group synchrony, and that reproductively mature fish were present at all sampling times. There were no significant temporal variations in the gonado‐somatic index ( I _G), hepato‐somatic index ( I _H), or plasma concentrations of Vtg, E₂ and T during the sampling period, which indicates that reproductive development is not synchronized within the population. Significant increases in I _G, I _H and plasma concentrations of Vtg, E₂ and T, however, were observed in vitellogenic fish, and in fish undergoing final maturation. A positive relationship was also found between the growth of oocytes and plasma concentrations of Vtg, E₂ and T, although the patterns of increase were different for each variable. Plasma concentrations of Vtg and E₂ rose steadily across oocyte sizes from 100 to 450 μm, but the rate of increase of plasma E₂ was slower than that of Vtg, and both reached a saturated concentration at oocyte sizes of c . 450 μm. In contrast, plasma concentrations of T showed no marked increase until oocytes grew beyond 400 μm.     

Gas exchange and metabolite fluctuations in green and yellow bands of variegated leaves of the monocotyledonous CAM species Agave americana

The variegated leaves of the Crassulacean acid metabolism (CAM) species Agave americana have a large central longitudinal green band with narrow yellow bands on either side. The yellow bands had 97% less pigment content, 84% lower ribulose‐1,5‐bisphosphate carboxylase/oxygenase activity, but only 20% lower phosphoenolpyruvate carboxylase activity than the green band. The green bands exhibited gas exchange typical of CAM plants, with most CO₂ uptake occurring at night, leading to a daily net CO₂ uptake of 127 mmol m⁻² day⁻¹. The yellow bands had some nighttime net CO₂ uptake but a larger loss during the daytime, indicating that they were sink tissues. Nocturnal citrate and malate accumulations for the yellow bands were 65 and 75%, respectively, of those of the green bands; sucrose supported 64‐83% of their nocturnal acid accumulation. This is the first evidence that agaves, which are malic‐enzyme‐type CAM plants, use sucrose as the carbon source for nocturnal acid accumulation. About 44% of the carbon demand of the yellow bands can be supplied by sucrose diffusing via the symplast from the adjacent green band, about 25% from fructose and glucose diffusion, and some via the apoplast.     

Interaction of gibberellins and phytochrome in the control of cowpea epicotyl elongation

The physiological response of cowpea ( Vigna sinensis L.) epicotyl explants to far‐red light (FR) and its interaction with gibberellins (GAs) have been investigated. The effect of FR and GA₁ varied with the age of the seedlings from which the explants were made: for FR, it decreased progressively with age (though the sensitivity of the epicotyls to FR did not change significantly until at least day 11), whereas it remained essentially constant for applied GA₁ between days 5 and 9 after sowing. This indicates that the loss of response to FR may be due to a decrease in endogenous GA levels in the epicotyl. For a range of GA₁ and GA₂₀ (0.01–1 µg explant⁻¹), both hormones were more active in FR than in R irradiated epicotyls, suggesting that phytochrome may affect GA sensitivity besides GA metabolism. The location of the epicotyl region most sensitive to FR (between 5 and 20 mm below the apex) was different from that to GAs (the upper 10 mm). Nevertheless, FR extended the region responsive to applied GAs, even in paclobutrazol‐treated epicotyls where elongation was due entirely to exogenous GAs. This means that modulation of epicotyl elongation by phytochrome, that occurs in a zone different from though overlapping with the GA‐sensitive subapical zone, is also mediated by GAs. Growth in the most FR‐sensitive region of the epicotyl stimulated by FR or GA₁ was due to cell elongation, and in the most GA‐sensitive region to both cell division and elongation. The effect of FR and GA₁ was negated by colchicine, indicating that microtubules may be involved in the response to both factors.     

Elevated pyruvate,orthophosphate dikinase (PPDK) activity alters carbon metabolism in C3 transgenic potatoes with a C4 maize PPDK gene

Changes in carbon metabolism and δ¹³C value of transgenic potato plants with a maize pyruvate,orthophosphate dikinase (PPDK; EC 2.7.9.1) gene are reported. PPDK catalyzes the formation of phospho enol pyruvate (PEP), the initial acceptor of CO₂ in the C₄ photosynthetic pathway. PPDK activities in the leases of transgenic potatoes were up to 5.4‐fold higher than those of control potato plants (wild‐type and treated control plants). In the transgenic potato plants, PPDK activity in leaves was negatively correlated with pyruvate content (r²= 0.81), and was positively correlated with malate content (r²= 0.88). A significant increase in the δ¹³C value was observed in the transgenic potato plants, suggesting a certain contribution of PEP carboxylase as the initial acceptor of atmospheric CO₂. These data suggest that elevated PPDK activity may alter carbon metabolism and lead to a partial operation of C₄‐type carbon metabolism. However, since parameters associated with CO₂ gas exchange were not affected, the altered carbon metabolism had only a small effect on the total photosynthetic characteristics of the transgenic plants.     

Peroxidase: Changes in soluble and bound forms during maturation and ripening of apples

Peroxidase (POD) enzymes, both soluble and bound, have been implicated in the ripening process of a number of fruits. However, their roles are poorly understood. The aim of this study was to define clearly the classes of POD activity in apple parenchyma tissues on the basis of their solubility in salt solutions, to determine the subcellular localisation of ionically‐bound POD activity, and to determine whether soluble‐ or ionically‐bound POD activities exhibit any relationships with maturation and ripening. The cortical parenchyma of apples ( Malus domestica Borkh cv. Cox's orange pippin) was investigated for changes in POD activity during maturation and post‐harvest storage at 0, 4, and 8°C at 100% relative humidity (RH), and controlled atmosphere (CA) storage (4°C in 1.25% O₂ and 0.5% CO₂). The parenchyma cells contained both soluble and ionically‐bound POD activity. The ionically‐bound POD was located in the cell wall in the middle lamella, plasmodesmata and cell corners, and could be extracted quantitatively with 0.4 M CaCl₂ at neutral pH to leave a POD‐free residue. During maturation, both forms of activity increased on a 'per apple' basis, but decreased as a function of fresh weight. Under all conditions of storage, soluble POD exhibited a peak of activity at approximately the mid‐point of ripening‐related softening; this event may constitute a biochemical marker. In contrast, ionically‐bound POD generally decreased during ripening, except under CA storage. CA storage inhibited ripening‐related softening after an initial loss of firmness. However, softening resumed after transfer of apples to normal atmosphere storage at 8°C. This may provide a new insight into controlling tissue softening.     

Thermoregulation, energy metabolism, and torpor in blossom-bats, Syconycteris australis (Megachiroptera)

Since little information is available on torpor in bats of the suborder Megachiroptera, we investigated whether the small (18 g) blossom-bat Syconycteris australis displays torpor in the laboratory. Bats entered daily torpor when food and water were withheld for one night and the air temperature (T_a) was below about 26°. Torpor began shortly after lights went on in the morning and lasted for a maximum of 12 hours. During torpor at T_a18°, metabolic rates fell to a minimum of about 15% of that in resting individuals at the same T_a, and to about 40% of the basal metabolic rate. The body temperature (T_b) during torpor was metabolically defended at or above about T_b 18°. Individuals that did not enter torpor in the morning reduced their T_b from about 34.5°, observed in resting individuals that had been fed during the previous night, to values between 30.2 and 32.8°, and the resting metabolic rate fell by about 25%. The ability to undergo short periods of torpor may explain why the distribution range of S. australis extends much further south than that of other small Australian megachiropteran bats.     

Metabolic depression and spleen and liver enlargement in juvenile Arctic charr Salvelinus alpinus exposed to chronic parasite infection

The present study on the connection between standard metabolic rate ( R _S) and chronic Diplostomum spp. infection resulted in a decrease in R _S, and an enlargement in spleen and liver sizes in the infected juvenile Arctic charr Salvelinus alpinus compared to control fish. As splenic enlargement observed in infected fish was not due to condition-related changes in the spleen, it could most probably be explained by increased leucocyte synthesis. The higher liver masses in infected S. alpinus may have been related to disorders in energetic function, which could have had major effects on biochemical regulation by the liver. The proposed metabolic syndrome with a possible reduction in insulin sensitivity in tissues results in ineffective glucose and lipid metabolism and thus it is suggested that chronic Diplostomum infection in S. alpinus might not impose direct energetic costs, but it may weaken the efficiency of energy metabolism and thus lead to lowered R _S.