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.     

Effects of feeding and induction strategy on the production of BmR1 antigen in recombinant E. coli

Aim:  To investigate the effects of feeding and induction strategies on the production of Bm R1 recombinant antigen.
Methods and Results:  Fed-batch fermentation was studied with respect to the specific growth rate and mode of induction to assess the growth potential of the bacteria in a bioreactor and to produce high yield of Bm R1 recombinant antigen. Cells were grown at a controlled specific growth rate (μ_set) during pre-induction, followed by constant feeding postinduction. The highest biomass (24·3 g l⁻¹) was obtained during fed-batch process operated at μ_set of 0·15 h⁻¹, whereby lower μ_set (0·075 h⁻¹) gave the highest protein production (9·82 mg l⁻¹). The yield of Bm R1 was increased by 1·2-fold upon induction with 1 mmol l⁻¹ IPTG (isopropyl-β- d -thiogalactoside) compared to using 5 mmol l⁻¹ and showed a further 3·5-fold increase when the culture was induced twice at the late log phase.
Conclusions:  Combination of feeding at a lower μ_set and twice induction with 1 mmol l⁻¹ IPTG yielded the best result of all variables tested, promising an improved method for Bm R1 production .
Significance and Impact of the Study:  This method can be used to increase the production scale of the Bm R1 recombinant antigen to meet the increasing demand for Brugia Rapid^™, a commercial diagnostic test for detection of brugian filariasis.     

The effects of body mass and feeding on metabolic rate in small juvenile Atlantic cod

Apparent specific dynamic action (SDA) amplitude in young juvenile Atlantic cod Gadus morhua (1 to 8 g wet mass), fed a standardized meal and then exercised in a circular swimming respirometer at a constant swimming speed of 0·5 ± 0·3 body lengths s^-1, occurred within l h after feeding in all juveniles. SDA amplitude were 1·4 to 1·8 times higher in fed fish compared to unfed fish, and rates of oxygen consumption decreased as body mass increased. SDA duration had a tendency to decrease with increasing body mass and was shortest, at 6 h, in the smallest fish (1–1·5 g), but increased to 10–11 h in the largest fish. Apparent SDA in fed fish ( R _r) scaled with a mass exponent of 0·89, while maximum metabolic rate ( R _max) determined by chasing fish to exhaustion and then measuring oxygen consumption for 12 h, and unfed routine metabolic rate (R_r) scaled with a mass exponent of 0·79 and 0·76 respectively. Relative aerobic scope ( R _max– unfed R _r) did not appear to vary over the 1 to 8 g increase in wet mass. These results show that as body mass increased in young juvenile Atlantic cod: (1) apparent SDA ( R _f) increased more rapidly than R _max, and (2) apparent SDA took up >98% of the relative aerobic scope and that young Atlantic cod allocated most of the energy to growth, and left little for other metabolic activities.     

Effect of ration size on growth and gross conversion efficiency of young lemon sharks, Negaprion brevirostris

Young lemon sharks, Negaprion brevirostris , were kept under controlled conditions in an aquarium and fed blue runner, Caranx crysos , at different ration levels. The relationship between feeding rate and growth rate was best described by a von Bertalanffy growth curve, which predicted a maximum growth rate of 140 kJ kg⁻¹ day⁻¹ (0·66% b.w. day⁻¹), a maintenance ration of 199 kJ kg⁻¹ day⁻¹ (1·06% b.w. day⁻¹), and losses due to starvation of -236kJ kg⁻¹ day⁻¹ (1·11% b.w. day⁻¹). The relationship between gross conversion efficiency ( K ₁) and feeding rate was also examined. K₁ ranged from - 64 to 25% and did not drop at high ration levels. Activity levels of both starved sharks and sharks fed at maintenance were not significantly different (0·2 body lengths s⁻¹). K ₁ values generated from both laboratory and field data suggest that young lemon sharks can convert food to new tissue as efficiently as teleosts.     

Effects of foraging mode and season on the energetics of the Marine Iguana, Amblyrhynchus cristatus

1. Marine Iguanas ( Amblyrhynchus cristatus ) inhabiting the rocky shores of the Galápagos Islands apply two foraging strategies, intertidal and subtidal foraging, in a seasonal climate. Effects of both foraging strategy and seasonality on the daily energy expenditure (DEE) were measured using doubly labelled water.
2. Difference in foraging mode did not result in significant differences in DEE.
3. On Santa Fé the DEE in the warm season was significantly higher than in the cool season (67·8 ± 21·8 kJ kg^–0·8 day^–1 vs 38·0 kJ kg^–0·8 day^–1). This difference can be explained by body temperature. A model estimate of the body temperature was used to predict monthly DEE figures, giving a year round budget. On average a 1-kg iguana would need only 47 kJ day^–1, or 17 mJ year ^–1. This is lower than previous estimates in which body temperatures were not taken into account.
4. The water flux of the Marine Iguana increases with increasing foraging time. The linear rise per minute foraging is roughly two times as high for subtidally foraging animals as for intertidal foragers.     

Effect of feeding level and feeding frequency on specific dynamic action in Silurus meridionalis

The effect of feeding level ( F _L; 0·5 to 4% dry diet mass per wet fish body mass) and feeding frequency (once every 4 days to twice per day) on postprandial metabolic response was investigated in southern catfish Silurus meridionalis at 27·5° C. The results showed that there was no significant difference in the specific dynamic action (SDA) coefficient among the groups of different feeding levels ( P  > 0·05). The duration increased from 26·0 to 40·0 h and the peak metabolic rate increased from 207·8 to 378·8 mg O₂ kg⁻¹ h⁻¹ when the feeding level was increased from 0·5 to 4%. The relationship between the peak metabolic rate ( R _P, mg O₂ kg⁻¹ h⁻¹) and F _L could be described as: R _P = 175·4 + 47·3 F _L( r ² = 0·943, n  = 40, P  < 0·001). The relationship between the SDA duration ( D , h) and F _L could be described as D =30·97 F _L^0·248 ( r ²=0·729, n =40, P  < 0·001).     

Plasma non-esterified fatty acid profiles in wild Atlantic salmon during their freshwater migration and spawning

Total plasma non-esterified fatty acid (NEFA) levels increased significantly in adult Atlantic salmon during the first months of their upstream migration and spawning in the Exploits River, Newfoundland, Canada. The highest levels occurred in May and were 5467±270·43 nmol ml⁻¹ for females and 4617±334·70 nmol ml⁻¹ for males. Significantly higher levels were maintained by females compared with males for most of the upstream migration. Between August and October, total plasma NEFA levels declined by 61% in females but only 23% in males. The decline in plasma monounsaturated and polyunsaturated fatty acid levels accounted for 74% of the loss of NEFAs in females. Specific plasma NEFAs such as 16: 0 (palmitic), 16: 1 (palmitoleic), 18: 1n9 (oleic) and 20: 5n3 (eicosapentaenoic acid) differed significantly between males and females during migration and spawning. The mean gonadosomatic index ( I _G) values of females in May and just prior to spawning were 0·37±0·01 and 10·25±0·32, respectively. The rapid decline in the plasma NEFA content of females coincided with the largest increase in their I _G (1·85±0·02–10·25±0·32). Corresponding I _G values for males were 0·34±0·01 in May and 3·33±0·78 prior to spawning. Plasma NEFA levels of spent salmon did not differ between sexes and were significantly lower than those of salmon preparing to spawn.     

Morphological and histological changes in the swim bladder during maturation of the Japanese eel

Silver eels Anguilla japonica ( I _G 1·5–3·5) had more developed rete mirabile, gas gland and submucosa than yellow eels ( I _G 0·4–1·5) and the development of swim bladder components increased with sexual maturity only in the early maturation process ( I _G 3·5). These observations indicate that the Japanese eel develops its swim bladder in either the river or shallow sea water and leaves for the open sea after the swim bladder has become adapted to a deeper-sea environment.     

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.     

Evaluation of rapid methods for the determination of okadaic acid in mussels

L.CROCI, A.STACCHINI, L.COZZI, G.CICCAGLIONI, F.MAZZEI, F.BOTRÈ AND L.TOTI. 2001 .
Aims: Two different screening methods, a Buffalo Green Monkey cytotoxicity test and a biosensor test, have been considered to replace the official mouse bioassay in monitoring for okadaic acid (OA) levels in mussels.
Methods and Results: Diarrhoetic shellfish poison-contaminated mussels from the Adriatic Sea were assayed in parallel by means of the mouse bioassay and both alternative methods. Both the cytotoxicity test and the biosensor test showed high sensitivity (OA 0·01 mg g^–1 hepatopancreas and 0·002 mg g^–1 hepatopancreas, respectively) and a high correlation with the mouse bioassay ( r =0·932, P  < 0·001 and r =− 0·850, P  < 0·001, respectively).
Conclusions: Both methods are efficacious, quick, inexpensive and provide data on the amount of toxin present in mussels.
Significance and Impact of the Study: Both methods, besides allowing the simultaneous assay of a great number of samples, comply with the ethical need to reduce the use of animals in the laboratory.     

Growth, diet and metabolism of common wolf–fish in the North Sea, a fast–growing population

The von Bertalanffy growth parameters for common wolf–fish Anarhichas lupus in the North Sea were: male: L ∞=111·2 cm, t ₀=–0·43 and K =0·12; and female: L ∞=115·1 cm, t ₀=–0·39 and K =0·11, making this the fastest growing stock reported. Resting metabolic rates (RMR±S.E.) and maximum metabolic rates (MMR±S.E.) for six adult common wolf–fish (mean weight, 1·39 kg) at 5° C were 12·18±1·6 mg O₂ kg^–1 h^–1 and 70·65±7·63 mg O₂ kg^–1 h^–1 respectively, and at 10° C were 25·43±1·31 mg O₂ kg^–1 h^–1 and 113·84±16·26 mg O₂ kg^–1 h^–1. Absolute metabolic scope was 53% greater at 10° C than at 5° C. The diet was dominated by Decapoda (39% overall by relative occurrence), Bivalvia (20%) and Gastropoda (12%). Sea urchins, typically of low energy value, occupied only 7% of the diet. The fast growth probably resulted from summer temperatures approximating to the optimum for food processing and growth, but may have been influenced by diet, and reduced competition following high fishing intensity.     

Lipid content and fatty acid composition of target tissues in wild Perca fluviatilis females in relation to hepatic status and gonad maturation

Variation in liver ultra‐structure and composition in relation to energy mobilization was investigated in female perch Perca fluviatilis from the Meuse River between August 2001 and June 2002. In April, just before spawning, the lipo‐somatic index ( I _F) was 0·3%, the gonado‐somatic index ( I _G) was 28% and the total lipid content of the liver was 2·53%. The average areas of lipid droplets and mitochondria were 0·05 and 0·06 μm², respectively. Glycogen supply reached 7·9% of the total area of the hepatocyte. During the sexual resting period, females accumulated energy in perivisceral fat and in the liver to reach 1·6% I _F and 4·85% of liver lipid content in August with lipid droplets average size of 0·09 μm² and glycogen average area of 15%. Liver cells contained a weakly developed rough endoplasmic reticulum (RER) and a great number of small mitochondria (average size 0·02 μm²). The I _G was 0·6% at this time. During the whole annual cycle, the average lipid content of female liver never exceeded 3·9 ± 1·9%. The concentration of docosahexaenoic (DHA), linolenic and linoleic acids increased in mature gonads while linolenic and linoleic acids decreased in the liver during the same period. Fatty acid composition of muscles of perch was characterized by a high content of DHA.     

Seasonal patterns of nitrogen fixation in termites

1. Termite nitrogenase activity was highest in autumn and spring (≈ 3 μg N₂ fixed termite fresh mass (g)^–1 day^–1) and lowest in winter and summer (≈ 0·8 μg N₂ fixed termite fresh mass (g)^–1 day^–1).
2. The nitrogenase activity of worker termites was significantly higher than all other castes (1·58 ± 0·27 μg N₂ fixed termite fresh mass (g)^–1 day^–1).
3. Worker termites constituted the largest proportion of all the castes throughout the study period (≈ 90%).
4. The localized input of fixed nitrogen by termites may reach 15·3 mg N log^–1 day^–1 and 5·6 g N log^–1 year^–1.     

Effect of water-borne zinc on osmoregulation in the freshwater amphipod Gammarus pulex (L.) from populations that differ in their sensitivity to metal stress

1. Although there is some evidence that exposure to heavy metals can disrupt osmoregulation in crustaceans, most studies have been carried out on relatively pollution-tolerant, marine or estuarine species. Consequently the effects of water-borne zinc (Zn) on osmoregulation by the freshwater amphipod, Gammarus pulex (L.), from two populations that differ in their heavy metal sensitivity, have been compared.
2. 'Clean' site animals (Clowne, Derbyshire) exhibited a marked haemoconcentration (after 4 days at 37·0 μmol Zn l^–1, 5 days at 18·2 μmol Zn l^–1) shown by an increase in haemolymph osmotic pressure (OP_h) and [Na⁺] and [K⁺]. However, after 5 days at 37·0 μmol Zn l^–1, haemolymph of survivors exhibited an OP_h significantly less than controls. 'Contaminated' site animals showed a reduction in OP_h (but not ions) only after 5 days at 76·2 μmol Zn l^–1.
3. There were differences in the threshold and nature of osmoregulatory response to Zn between animals from 'clean' and 'contaminated' sites, but only at concentrations in excess of those (a) known to affect growth and reproduction in 'clean' site animals and (b) occurring at the 'contaminated' site. Clearly population differences in physiological capacity and tolerance do exist but their ecological significance is unclear.     

Body temperature and insulation in diving Great Cormorants and European Shags

1. Cormorants are typically considered as wettable diving birds with high thermoregulatory costs and are presumed to exert substantial predatory pressure on fish stocks.
2. The stomach temperatures of seven Great Cormorants and three European Shags were recorded during a total of 108 foraging trips undertaken near the Chausey Islands breeding colony (France).
3. Both species kept a constant body temperature during the dive series which lasted up to 158 min and were conducted in 12°C water. Consequently, assuming that heat loss to the water is equal to heat production in diving Great Cormorants, the minimal insulating plumage air volume was calculated to be 0·371 × 10^–3 m³ (corresponding to a 1·62-mm air layer) in males and 0·347 × 10^–3 m³ (corresponding to a 1·90-mm air layer) in females.
4. Furthermore, it is shown that plumage air volume and dive depth are the major factors influencing heat flux to the water and that the energetics of diving Great Cormorants may also vary substantially according to fat layer thickness, water temperature and body temperature. Swim speed plays only a minor role.
5. Considering these results, it is postulated that Great Cormorants may have optimized plumage air volume so as to minimize both mechanical costs (upthrust) and thermoregulatory costs of swimming in cold, shallow water.
6. Finally, body temperature patterns recorded in different cormorant species while diving are compared.     

Sibling cannibalism among juvenile vundu under controlled conditions. II. Effect of body weight and environmental variables on the periodicity and intensity of type II cannibalism

Cannibalism among starved groups of juvenile (19–48 days old) vundu catfish Heterobranchus longifilis was 66·5% nocturnal, and its impact under modified day length was proportional to the duration of the dark phase. Shallow depth and high population density decreased the intensity of cannibalism, whereas low density and deeper environments had an opposite effect. The presence of refuges had no significant effect on cannibalism. The maintenance ( R _maint) and maximum ( R _max) daily food rations (% day⁻¹) of cannibals feeding on live prey were modelled as R _maint=3·899 W _C^0·327 ( r ²=0·684; d.f.=31), and R _max=49.545 W _C^0·321 ( r ²=0·999; d.f.=5), where W _C was the body weight of the cannibal (g). The latter model indicated that the impact of a cannibal on a population decreased by a 20% margin each time the cannibal doubled its body weight, and suggested that cannibalism among vundu would become insignificant for cannibals heavier than 30 g. The significance of these findings is discussed within the contexts of vundu aquaculture and of general, conceptual models of the dynamics of cannibalism among fishes.     

Cardiorespiratory status of triploid brown trout during swimming at two acclimation temperatures

At 14° C, standard metabolic rate (75·1 mg O₂ h⁻¹ kg⁻¹), routine metabolic rate (108.8 mg O₂ h⁻¹ kg⁻¹), active metabolic rate ( c . 380 mg O₂ h⁻¹ kg⁻¹), critical swimming speed (U_crit 1·7 BL s⁻¹), heart rate 47 min⁻¹), dorsal aortic pressure (3·2 kPa) and ventilation frequency (63 min⁻¹) for triploid brown trout Salmo trutta were within the ranges reported for diploid brown trout and other salmonids at the same temperature. During prolonged swimming ( c . 80% U _crit), cardiac output increased by 2·3-fold due to increases in heart rate (1·8-fold) and stroke volume (1·2-fold). At 18° C, although standard and routine metabolic rates, as well as resting heart rate and ventilation frequency increased significantly, active metabolic rate and certain cardiorespiratory variables during exercise did not differ from those values for fish acclimated to 14° C. As a result, factorial metabolic scope was reduced (2·93-fold at 18° C v . 5·13-fold at 14° C). Therefore, it is concluded that cardiorespiratory performance in triploid brown trout was not unusual at 18° C, but that reduced factorial metabolic scope may be a contributing factor to the mortality observed in triploid brown trout at temperatures near 18° C.     

The linamarase of Mucor circinelloides LU M40 and its detoxifying activity on cassava

Mucor circinelloides LU M40 produced 12·2 mU ml⁻¹ of linamarase activity when grown in a 3 l fermenter in the following optimized medium (g l⁻¹ deionized water): pectin, 10·0; (NH₄)₂SO₄,
1·0; KH₂PO₄, 2·0; Na₂HPO₄, 0·7; MgSO₄.7H₂O, 0·5; yeast extract, 1·0; Tween-80,
1·0, added after 48 h of fermentation. The purified linamarase was a dimeric protein with a molecular mass of 210 kDa; the enzyme showed optimum catalytic activity at pH 5·5 and 40 °C and had a wide range (3·0–7·0) of pH stability. The enzyme substrate specificity on plant cyanogenic glycosides was wide; the K_m value for linamarin was 2·93 mmol l⁻¹. The addition, before processing, of the fungal crude enzyme to cassava roots facilitated and shortened detoxification; after 24 h of fermentation, all cyanogenic glycosides were hydrolysed.     

Modelling the growth of Weissella cibaria as a function of fermentation conditions

Aims:  To investigate the effect of pH, water activity ( a _w) and temperature on the growth of Weissella cibaria DBPZ1006, a lactic acid bacterium isolated from sourdoughs.
Methods and Results:  The kinetics of growth of W. cibaria DBPZ1006 was investigated during batch fermentations as a function of pH (4·0–8·0), a _w (0·935–0·994) and temperature (10–45°C) in a rich medium. The growth curve parameters (lag time, growth rate and asymptote) were estimated using the dynamic model of Baranyi and Roberts (1994. A dynamic approach to predicting bacterial growth in food. Int J Food Microbiol 23, 277–294). The effect of pH, a _w and temperature on maximum specific growth rate (μ_max) were estimated by fitting a cardinal model. μ_max under optimal conditions (pH = 6·6, a _w = 0·994, T  = 36·3°C) was estimated to be 0·93 h⁻¹. Minimum and maximum estimated pH and temperature for growth were 3·6 and 8·15, and 9·0°C and 47·8°C, respectively, while minimum a _w was 0·918 (equivalent to 12·2% w/v NaCl).
Conclusions:  Weissella cibaria DBPZ1006 is a fast-growing heterofermentative strain, which could be used in a mixed starter culture for making bread.
Significance and Impact of the Study:  This is the first study reporting the modelling of the growth of W. cibaria , a species that is increasingly being used as a starter in sourdough and vegetable fermentations.     

Do plant species with high relative growth rates have poorer chemical defences?

1. Most theories of plant strategies assume the presence of certain 'trade-offs'. One such evolutionary trade-off assumes a decrease in growth rate with increasing investment in chemical defences in species adapted to different levels of habitat fertility.
2. To test this hypothesis, we grew 31 herbaceous species of Asteraceae under controlled conditions of temperature (25 °C), humidity (80%), light (500 μmol m^–2 s^–1) and photoperiod (16 h day^–1) in a modified Hoagland hydroponic solution. The plants grew from seed for 35 days post-germination and were harvested at 14, 21, 28 and 35 days. Relative growth rate (RGR) was calculated as well as a general measure of potential phytochemical toxicity (LC₅₀) using an alcohol extraction of secondary compounds followed by Brine Shrimp bioassay and an assay of total phenolics.
3. The interspecific correlation between RGR and the potential phytochemical toxicity was weak and non-significant ( r _S = 0·12, P = 0·53). The correlation between RGR and total phenolics was weak, positive but significant ( r _S = 0·40, P = 0·03).
4. These results suggest that such an evolutionary trade-off does not exist in this group of Asteraceae.