Metabolic properties of Northern krill, Meganyctiphanes norvegica, from different climatic zones. II. Enzyme characteristics and activities

Activities and characteristics of two metabolic key enzymes, citrate synthase (CS) and pyruvate kinase (PK), were studied in the Northern krill, Meganyctiphanes norvegica, with respect to adaptive properties under different thermal conditions. Krill were sampled during late winter/spring and summer from the constantly warm Ligurian Sea (12-13°C below the thermocline), the colder but also comparatively constant Clyde Sea (7-8°C), and the variable Kattegat (2-16°C). Both enzymes showed distinct tissue- and organ-specific activities, which were highest in the pleopods - the principal locomotive organs. The fourth and fifth abdominal segments, however, were used for routine investigation due to lowest variability. Specific activity of CS and PK did not differ between seasons in krill from the Kattegat or the Clyde Sea. In the Ligurian Sea, in contrast, specific CS activities were significantly lower during summer. Analysis of individual data illustrated a decrease of CS activity with size and an increase of PK activity with size. Taking these allometric effects into account, as emphasized by calculating the ratio between both enzymes, variation of CS and PK activities turned out to be solely dependent on body size, which differed between locations and seasons. Ligurian krill from the summer, however, were unique in that they showed a lower CS/PK ratio than would be predicted by the scaling effect. Thermal characteristics of each enzyme were similar between locations and seasons. During the winter, in Kattegat and Clyde Sea krill, K_m values (Michaelis-Menten constant) of CS towards acetyl-coenzyme A exhibited an almost constant level over the experimental temperature range of 4-16°C. During summer, however, K_m values were lower at 8°C in the Clyde Sea and at 12°C in the Kattegat. In Ligurian krill from the summer, K_m values were consistently lower than those of winter krill over the entire experimental temperature range. In conclusion, Kattegat and Clyde Sea krill show only minor adaptations to their respective thermal environments in terms of CS and PK characteristics. Ligurian krill, in contrast, exhibited decreased specific CS activity during summer, which might be compensated by elevated enzyme-substrate affinity as indicated by lower K_m values. Since temperature was constant during both seasons, this effect cannot be explained as a reaction to thermal conditions. Consequently, oligotrophic conditions in the Ligurian Sea during summer may entail a reduction in the somatic performance of krill, which is reflected by lower CS activity.     

Ovarian development and spawning in relation to the moult cycle in Northern krill, Meganyctiphanes norvegica (Crustacea: Euphausiacea), along a climatic gradient

Adaptive processes linked to reproduction were studied comparatively for three populations of Northern krill, Meganyctiphanes norvegica (M. Sars, 1857), sampled during winter and summer cruises in the Clyde Sea (W Scotland), the Kattegat (E Denmark), and the Ligurian Sea (NW Mediterranean). The aim was to investigate the functional relationship between egg production and moulting under contrasted climatic and environmental conditions. A staging system for female sexual development established for live krill was complemented by a histological study of the ovary at various developmental steps. During the reproductive season, all adult female krill were engaged in cyclical egg production. During experiments, female krill released one batch of mature oocytes in one or two spawning events. The ovary of postspawn female krill still contained developing oocytes for another egg batch. In the non-reproductive period, all female krill had a resting ovary. Ovarian structure and pattern of egg production were identical in the three populations, but seasonal timing of egg production was different. The model proposed for the Ligurian population of the annual cycle of ovarian development can be extended to the other two populations, taking into account the seasonal characteristics of each site. Random field samples were staged simultaneously for moult cycle and for sexual development. Moult stages and the seasonal variation of the intermoult period were studied for the Kattegat population using multi-year data and compared to data obtained during summer/winter cruises in the Clyde and the Ligurian Sea. At the three sites, intermoult period was shorter and temperature-dependent during the reproductive period, concurrent with the season of greatest food availability. During most of the year and the period of sexual rest, moulting activity was reduced. The relationship between spawning and the moult cycle was studied comparatively for the three populations. Eggs were released during the premoult phase of a “spawning moult cycle”, in one or two spawnings associated with apolysis and Moult Stage D1, respectively. Yolk accumulation for the next egg batch was completed during an alternating “vitellogenic moult cycle”. A model for the timing of cyclical egg production in relation to moulting, as proposed for the Kattegat, can be extended to the other populations, taking into account intermoult period variation with temperature. Temperature appeared to be the principal environmental factor controlling growth (through moulting) and egg production during the reproductive season, in connection with favourable trophic conditions. Received: 22 December 1997 / Accepted: 29 August 1998     

Temperature effects on the metabolism of larvae of the Antarctic starfish Odontaster validus, using a novel micro-respirometry method

Oxygen consumption of individual larvae of the Antarctic sea-star Odontaster validus was measured during the 50-day period following fertilisation. Values ranged from 0.76 pmol O₂ h^-1 for one specimen at the coeloblastula stage to 77.6 pmol O₂ h^-1 for one bipinnaria larva. At 0°C the mean oxygen consumption rate of an individual larva increased from 10.9 pmol O₂ h^-1 (standard error of the mean, SEM, 0.13) for a gastrula larva, 13 days post-fertilisation, to 25.4 pmol O₂ h^-1 (SEM 3.5) at the bipinnaria stage (50 days post-fertilisation). Gastrulae reared at -0.5°C did not have significantly different oxygen consumption rates between days 13 and 45 post-fertilisation (mean=11.4 pmol O₂ h^-1). Individual metabolic rates were highly variable, covering more than a 40-fold range. At 2°C gastrula oxygen consumption was on average 45% higher (17.35 pmol O₂ h^-1), giving a Q₁₀ temperature effect of 4.4. For bipinnaria, mean oxygen consumption in 2°C larvae (31.4 pmol O₂ h^-1) was not significantly different from that in larvae at -0.5°C, suggesting bipinnaria metabolism may be less sensitive to temperature change than earlier stages. At 2°C the bipinnaria stage was reached at 30-35 days compared with 45-50 days at 0°C, giving a Q₁₀ of 4.5 for temperature effects on development. The method here used a new, highly sensitive micro-respirometry method that is inexpensive and straightforward in design. Individual larvae of O. validus were held in 35- to 50-µl respirometers. These larvae have very low metabolic rates, and published work on such organisms have utilised at least 25 individuals per chamber. The oxygen content of the respirometers was measured using a 25-µl sample injected into a couloximeter. Oxygen consumption rates down to -1 pmol h^-1 can be detected. Under optimum conditions oxygen consumption of a single larva of -4 pmol O₂ h^-1 was measured with an accuracy of ᆨ%. Values of ~15 pmol h^-1 could routinely be measured with this accuracy. This method would allow oxygen consumption to be evaluated in individual field-caught larvae of most marine ectotherms.     

Metabolic rates of juvenile scalloped hammerhead sharks (Sphyrna lewini)

Oxygen consumption of juvenile scalloped hammerhead sharks, Sphyrna lewini, was measured in a Brett-type flume (volume=635 l) to quantify metabolic rates over a range of aerobic swimming speeds and water temperatures. Oxygen consumption (log transformed) increased at a linear rate with increases in tailbeat frequency and swimming speed. Estimates of standard metabolic rate ranged between 161 mg O₂ kg^-1 h^-1 at 21°C and 203 mg O₂ kg^-1 h^-1 at 29°C (mean-SD: 189ᆣ mg O₂ kg^-1 h^-1 at 26°C). Total metabolic rates ranged from 275 mg O₂ kg^-1 h^-1 at swimming speeds of 0.5 body lengths per second (L s^-1) to a maximum aerobic metabolic rate of 501 mg O₂ kg^-1 h^-1 at 1.4 L s^-1. Net cost of transport was highest at slower swimming speeds (0.5-0.6 L s^-1) and was lowest between 0.75 and 0.9 L s^-1. Therefore, these sharks are most energy efficient at swimming speeds between 0.75 and 0.9 L s^-1. These data indicate that tailbeat frequency and swimming speed can be used as predictors of metabolic rate of free-swimming juvenile hammerhead sharks.     

Response of zooxanthellae in symbiosis with the Mediterranean corals Cladocora caespitosa and Oculina patagonica to elevated temperatures

Scleractinian symbiotic corals living in the Ligurian Sea (NW Mediterranean Sea) have experienced warm summers during the last decade, with temperatures rapidly increasing, within a few days, to 3–4°C above the mean value of 24°C. The effect of elevated temperatures on the photosynthetic efficiency of zooxanthellae in symbiosis with temperate corals has not been well investigated. In this study, the corals, Cladocora caespitosa and Oculina patagonica were collected in the Ligurian Sea (44°N, 9°E), maintained during 2 weeks at the mean summer temperature of 24°C and then exposed during 48 h to temperatures of 24 (control), 27, 29 and 32°C. Chlorophyll (chl) fluorescence parameters [F _v/F _m, electron transport rate (ETR), non-photochemical quenching (NPQ)] were measured using pulse amplitude modulated (PAM) fluorimetry before, during the thermal increase, and after 1 and 7 days of recovery (corals maintained at 24°C). Zooxanthellae showed a broad tolerance to temperature increase, since their density remained unchanged and there was no significant reduction in their maximum quantum yield (F _v/F _m) or ETR up to 29°C. This temperature corresponded to a 5°C increase compared to the mean summer temperature (24°C) in the Ligurian Sea. At 32°C, there was a significant decrease in chl contents for both corals. This decrease was due to a reduction in the chl/zooxanthellae content. For C. caespitosa, there was also a decrease in ETR_max, not associated with a change in F _v/F _m or in the non-photochemical quenching (NPQ); for O. patagonica, both ETR_max and F _v/F _m significantly decreased, and NPQ_max showed a significant increase. Damages to the photosystem II appeared to be reversible in both corals, since F _v/F _m values returned to normal after 1 day at 24°C. Zooxanthellae in symbiosis with the Mediterranean corals investigated can therefore be considered as resistant to short-term increases in temperature, even well above the maximum temperatures experienced by these corals in summer.     

Biomass, photosynthesis and growth of Laminaria saccharina in a high-arctic fjord, NE Greenland

Biomass, photosynthesis and growth of the large, perennial brown alga Laminaria saccharina (L.) Lamour. were examined along a depth gradient in a high-arctic fjord, Young Sound, NE Greenland (74°18'N; 20°14'W), in order to evaluate how well the species is adapted to the extreme climatic conditions. The area is covered by up to 1.6-m-thick ice during 10 months of the year, and bottom water temperature is <0°C all year round. L. saccharina occurred from 2.5 m depth to a lower depth limit of about 20 m receiving 0.7% of surface irradiance. Specimen density and biomass were low, likely, because of heavy ice scouring in shallow water and intensive feeding activity from walruses in deeper areas. The largest specimens were >4 m long and older than 4 years. In contrast to temperate stands of L. saccharina, old leaf blades (2-3 years old) remained attached to the new blades. The old tissues maintained their photosynthetic capacity thereby contributing importantly to algal carbon balance. The photosynthetic characteristics of new tissues reflected a high capacity for adaptation to different light regimes. At low light under ice, or in deep water, the chlorophyll a content and photosynthetic efficiency (!) were high, while light compensation (E_c) and saturation (E_k) points were low. An E_c of 2.0 µmol photons m^-2 s^-1 under ice allowed photosynthesis to almost balance, and sometimes exceed, respiratory costs during the period with thick ice cover but high surface irradiance, from April through July. Rates of respiration were lower than usually found for macroalgae. Annual elongation rates of leaf blades (70-90 cm) were only slightly lower than for temperate L. saccharina, but specific growth rates (0.48-0.58 year^-1) were substantially lower, because the old blades remained attached. L. saccharina comprised between 5% and 10% of total macroalgal biomass in the area, and the annual contribution to primary production was only between 0.1 and 1.6 g C m^-2 year^-1.     

On the timing of moulting processes in reproductively active Northern krill Meganyctiphanes norvegica

The interactions between moult phasing, growth and environmental cues in Northern krill (Meganyctiphanes norvegica) were examined through analysing populations at seasonal, weekly, and daily timescales. The analyses were carried out on resident populations of krill found in three different neritic locations that experience similar environmental signals (the Clyde Sea, Scotland; the Kattegat, Denmark; Gullmarsfjord, Sweden). Seasonal analyses were carried out on the Clyde Sea population and showed that moulting frequency increased significantly moving from winter to summer. The proportion of moulting females in summer samples was often more than double the proportion of moulting males, suggesting that females had a comparatively shorter intermoult period (IMP). Weekly samples taken from the Kattegat showed a similar pattern. However, although the difference between the proportion of female and male moulters was significant in one week, it was not another, mainly because of the variability in the proportion of female moulters. Such variability in females was equally evident in the daily samples taken at Gullmarsfjord. It suggests that females have a shorter IMP (12.5 days) than males (18.4 days) and are more likely to moult in synchrony. Nevertheless, the daily samples revealed that males are also capable of moult synchronisation, although less frequently than females. Shortened IMPs in females were not a result of the abbreviation of specific moult stages. Accordingly, reproductive activity did not alter the course of the normal moult cycle. There was no significant difference between the total body lengths of males and females indicating that females achieve the same levels of growth despite moulting more frequently and having to provision the energy-rich ovaries. This is in contrast to most other crustaceans where the energy costs of reproduction reduce female growth. The fact that females were less abundant than males, probably by suffering a greater level of mortality, suggests that different behavioural strategies, particularly vertical migration regimes, were adopted by each sex to maximise growth and reproduction.     

Large-scale oceanic distribution and population structure of Calanus finmarchicus, in relation to physical environment, food and predators

The investigation was carried out from 62°N to 73°N and from 14°E to 11°W in the Norwegian Sea during 19 June-12 July 1997. Regional differences in the phase of the seasonal development of the plankton community were evident, most pronounced across the Arctic front. In the Coastal and eastern Atlantic domains, post-bloom conditions prevailed, characterised by low chlorophyll a (chl a) levels and a phytoplankton assemblage dominated by coccolithophorids and small flagellates. During the study period, egg production rates of Calanus finmarchicus were low (<10 eggs female^-1 day^-1), older copepodite stages dominated, and the seasonal descent to deeper waters had started. In the Arctic domain, bloom conditions were evident by high chl a levels and a high abundance of large diatoms. Egg production rates were higher (a maximum of 29 eggs female^-1 day^-1), but the dominance of stages CI-CIII indicated that considerable spawning had already occurred prior to the spring bloom. The seasonal descent had barely started. Both invertebrate and fish predators were most abundant in the Coastal and eastern Atlantic domains, with abundance strongly decreasing north-westwards. No tight relationship between total abundance of invertebrate or fish predators and that of C. finmarchicus was apparent. However, a weak, but significant, relationship between abundance of young stages of chaetognaths and Euchaeta spp. versus young stages of C. finmarchicus was found, indicating that these invertebrate predators develop parallel to the development of the new cohort of C. finmarchicus. In early summer, C. finmarchicus had reached overwintering stages, and had started to accumulate in deeper waters in areas with the highest abundance of horizontally migratory planktivorous fish.     

Determination of lethal dissolved oxygen levels for selected marine and estuarine fishes, crustaceans, and a bivalve

The objective of this study was to provide a database of the incipient lethal concentrations for reduced dissolved oxygen (DO) for selected marine and estuarine species including 12 species of fish, 9 crustaceans, and 1 bivalve. All species occur in the Virginian Province, USA, which is a cold temperate region. The study period was August 1987 to September 1995. Standard bioassay procedures were employed, with most tests being of 4-day duration. Up to eight lethal concentrations (LCs) between LC₀₅ and LC₉₅ were estimated. The study provides four general conclusions about determining lethal thresholds of low DO for these organisms. First, the concentration response curve of most species did not change greatly beyond day 1 of the exposure with the exception of crustacean larvae, which were usually more sensitive on day 4, possibly due to molting. Second, acute LC₅₀ values (1- to 4-day) for low DO were influenced by life-stage and habitat, with pelagic larvae generally being the most sensitive and benthic juveniles the least. Species mean LC₅₀ values ranged from 1.4 to 3.3 mg l^-1 for larvae, 1.0 to 2.2 mg l^-1 for postlarvae, and 0.5 to 1.6 mg l^-1 for juveniles. No intraspecific differences in LC₅₀ were detected between larval stages in crustaceans or with age in larval fishes. The response range between LC₀₅ and LC₉₅ was narrowest for the least sensitive organisms (0.6 mg l^-1), and broadened with sensitivity. The mean LC₁₀:LC₅₀ ratio for all species was 1.32 for larvae and juveniles, and 1.36 for postlarvae. The ratio for postlarvae represents only four species, and hence is not considered different from the other life stages. Third, variability increased with increased species and life stage sensitivity to low DO, and with endpoints of LC₁₅ and below, which reduces the certainty of some of these results. Lastly, no influence of temperatures between 20°C and 30°C was detected in a small set of tests with thermally acclimated crustacean larvae. This data set has been used to describe protection limits for juvenile and adult survival, and for larval recruitment for the case of persistent (₄ h) low DO for estuarine and coastal waters of the Virginian Province, USA.     

Influence of environmental factors on burrow irrigation and oxygen consumption in the mudflat invertebrate Urechis caupo

We examined burrow irrigation activity by the mudflat worm Urechis caupo in response to suspended food, ambient hypoxia (down to 3.3 kPa PO₂), hydrogen sulfide exposure (up to 100 µmol l^-1), and short-term temperature change (range 10-22°C). In normoxic, nutrient-free water at 14°C, O₂ consumption ( [(M)\dot]O₂ ) \left( {\dot M{\rm O}_2 } \right) was 45 nmol min^-1 g^-1, water flow rate ( [(V)\dot]_W ) \left( {\dot V_{\rm W} } \right) was 27 ml min^-1 (0.66 ml min^-1 g^-1), frequency of peristaltic waves (F_P) was 2.6 contractions min^-1, stroke volume (SV) was 11 ml, and O₂ extraction coefficient (EO₂) was 0.27. Adding suspended food to the burrow water occasionally elicited stereotypical feeding behavior but had no effect on any measured variables during nonfeeding periods. Hypoxia greatly decreased [(M)\dot]O₂ \dot M{\rm O}_2 (75% reduction at 3.3 kPa PO₂) but did not affect [(V)\dot]_W \dot V_{\rm W} , F_P, SV, or EO₂. Sulfide at 50 µmol l^-1 or less had no effect on burrow irrigation activity, whereas 100 µmol l^-1 sulfide decreased [(V)\dot]_W \dot V_{\rm W} by 58% and F_P by 50% but had no effect on SV. Temperature strongly affected [(V)\dot]_W \dot V_{\rm W} (Q₁₀ of 1.9 from 10°C to 22°C). We propose that U. caupo's ability to live in the hypoxic, sulfidic mud of productive mudflat environments, combined with its very efficient mucous net, allows it to process much less water for feeding than other suspension-feeding invertebrates. This, in turn, necessitates an efficient O₂ extraction mechanism, which is provided by the water lung activity of U. caupo's unique hindgut.     

Oxygen and carbon stable isotopic profiles of the fan mussel, Pinna nobilis, and reconstruction of sea surface temperatures in the Mediterranean

Stable oxygen and carbon isotope profiles ('¹⁸O_skeletal and '¹³C_skeletal), taken along the direction of growth from the umbo to the shell margin in shells of the pinnid Pinna nobilis, were used to reconstruct sea surface temperatures (SST) in the south-east Mediterranean and ontogenetic records of metabolic CO₂ incorporation. Comparison of the seasonal cycle of SST, predicted from the '¹⁸O_skeletal record of a small (young) rapidly growing pinnid and temperature measured with a continuous in situ recorder showed that P. nobilis calcifies under isotopic equilibrium with surrounding seawater, thus indicating that P. nobilis shells can be used as a reliable predictor of SST. A 10-year SST record for the south-east Mediterranean was reconstructed from the shell profiles of four pinnid shells of different sizes and ages collected in 1995 and 1996. Reliable resolution of the seasonal SST could only be achieved during the first 4 years of shell growth. As the pinnids grew older, the temperature record was poorly resolved because the shell growth had diminished with age, resulting in time-averaging of the record. The amplitude of the generated seasonal temperature cycle compared favourably (DŽ°C) with a long-term temperature record from northern Mediterranean waters. Clear seasonal cycles in '¹³C_skeletal were observed with an amplitude of ~1.0‰, similar to the calculated seasonal changes in '¹³C of seawater (0.6‰) overlying seagrass meadows. An ontogenetic trend towards less positive '¹³C_skeletal values was too large to be attributed to any decrease in '¹³C in seawater resulting from the invasion of anthropogenic CO₂. It is suggested that the temporal changes of '¹³C_skeletal are due to incorporation of respiratory CO₂ into the extrapallial fluid and reflect changes in the metabolic activity of the pinnid rather than changes in the isotopic composition of dissolved inorganic carbon within the surrounding seawater.     

Metabolic rates of epipelagic marine copepods as a function of body mass and temperature

Metabolic rates (oxygen consumption, ammonia excretion, phosphate excretion) have been calculated as a function of body mass (dry, carbon, nitrogen and phosphorus weights) and habitat temperature, using multiple regression. The metabolic data used for this analysis were species structured, collected from Arctic to Antarctic seas (temperature range: -1.7°C to 29.0°C). The data were further divided into geographical and/or seasonal groups (35 species and 43 data sets for oxygen consumption; 38 species and 58 data sets for ammonia excretion; 22 species and 31 data sets for phosphate excretion). The results revealed that the variance attributed to body mass and temperature was highest (93-96%) for oxygen consumption rates, followed by ammonia excretion rates (74-80%) and phosphate excretion rates (46-56%). Among the various body mass units, the best correlation was provided by the nitrogen unit, followed by the dry weight unit. The calculated Q₁₀ values varied slightly according to the choice of body mass units; overall ranges were 1.8-2.1 for oxygen consumption rates, 1.8-2.0 for ammonia excretion rates and 1.6-1.9 for phosphate excretion rates. The effects of body mass and temperature on the metabolic quotients (O:N, N:P, O:P) were insignificant in most cases. Although the copepod metabolic data used in the present analysis were for adult and pre-adult stages, possible applications of the resultant regression equations to predict the metabolic rates of naupliar and early copepodite stages are discussed. Finally, global patterns of net growth efficiency [growth (growth+metabolism)^-1] of copepods were deduced by combining the present metabolic equation with Hirst and Lampitt's global growth equation for epipelagic marine copepods.     

Temperature acclimation of upper and lower thermal limits and freeze resistance in the nonindigenous brown mussel, Perna perna (L.), from the Gulf of Mexico

Acute and chronic upper and lower thermal limits and freeze resistance were investigated in the nonindigenous brown mussel, Perna perna, from the Texas Gulf of Mexico coast in order to assess its potential distribution in North American coastal waters. This species' long-term, incipient lower and upper thermal limits were 7.5°C and 30°C, congruent with the seasonal ambient water temperature range of 10-30°C reported for other populations worldwide. Effects of temperature acclimation and individual size on survival time were most pronounced on chronic exposures to lethal temperatures approaching incipient lower or upper thermal limits. When exposed to temperature increasing at 0.1°C min^-1, the acute upper lethal limit was 44°C regardless of acclimation temperature or individual size. P. perna had a limited freeze resistance, being intolerant of emersion at -2.5°C. This species' narrow incipient thermal limits, limited capacity for temperature acclimation and poor freeze resistance may account for its restriction to subtidal and lower eulittoral zones of cooler subtropical rocky shores. Near extinction of P. perna from Texas Gulf of Mexico waters occurred in the summer of 1997 when mean surface-water temperatures approached its incipient upper limit of 30°C.     

Characterization of urease activity in three marine phytoplankton species,<Emphasis Type="Italic"> Aureococcus anophagefferens</Emphasis>,<Emphasis Type="Italic"> Prorocentrum minimum</Emphasis>, and<Emphasis Type="Italic"> Thalassiosira weissflogii</Emphasis>

The availability of different forms of nitrogen in coastal and estuarine waters may be important in determining the abundance and productivity of different phytoplankton species. Although urea has been shown to contribute as much as 50% of the nitrogen for phytoplankton nutrition, relatively little is known of the activity and expression of urease in phytoplankton. Using an in vitro enzyme assay, urease activities were examined in laboratory cultures of three species: Aureococcus anophagefferens Hargraves et Sieburth, Prorocentrum minimum (Pavillard) Schiller, and Thalassiosira weissflogii (Grunow) Fryxell et Hasle. Cultures of P. minimum and T. weissflogii were grown on three nitrogen sources (NO₃^m, NH₄⁺, and urea), while A. anophagefferens was grown only on NO₃^m and urea. Urease was found to be constitutive in all cultures, but activity varied with growth rate and assay temperature for the different cultures. For A. anophagefferens, urease activity varied positively with growth rate regardless of the N source, while for P. minimum, urease activity varied positively with growth rate only for cultures grown on urea and NH₄⁺. In contrast, for T. weissflogii, activity did not vary with growth rate for any of the N sources. For all species, urease activity increased with assay temperature, but with different apparent temperature optima. For A. anophagefferens, in vitro activity increased from near 0-30°C, and remained stable to 50°C, while for P. minimum, increased in vitro activity was noted from near 0-20°C, but constant activity was observed between 20°C and 50°C. For T. weissfloggii, while activity also increased from 0°C to 20°C, subsequent decreases were noted when temperature was elevated above 20°C. Urease activity had a half-saturation constant of 120-165 wg atom N l^у in all three species. On both an hourly and daily basis, urease activity in A. anophagefferens exceeded nitrogen demand for growth. In P. minimum, urease activity on an hourly basis matched the nitrogen demand, but was less than the demand on a daily basis. For T. weissflogii, urease activity was always less than the nitrogen demand. These patterns in urease activity in three different species demonstrate that while apparently constitutive, the regulation of activity was substantially different in the diatom. These differences in the physiological regulation of urease activity, as well as other enzymes, may play a role in their ecological success in different environments.     

Combined effect of temperature and food concentration on the grazing rate of the rotifer Brachionus plicatilis

We developed a predictive relationship to determine the grazing rate of Brachionus plicatilis at given temperatures and food concentrations; this function could be applied to experimental culturing and aquaculture practices. Grazing experiments were conducted at temperatures between 5°C and 40°C and at food concentrations, of the flagellate Isochrysis galbana, ranging between ~0 and 10⁶ ml^-1. In total, 136 grazing rates were determined, using the prey depletion method, for rotifers acclimated to treatments for 0.5 or 4 h. The response of grazing rate to temperature and food concentration was described using a model that combined a rectangular hyperbolic function for food concentration and a sigmoidal function for temperature. Using non-linear curve-fitting methods an equation was obtained: G=(452F)/(159000+F)Ǵ.94/(1+2190002T^-4.35) , where G is the grazing rate (flagellates rotifer^-1 min^-1), F is the food concentration (flagellates ml^-1), and T is temperature (°C). The equation indicates a maximum grazing rate of ~35 prey rotifer^-1 min^-1, above ~4᎒⁵ prey ml^-1 and 25°C.     

Effects of emersion and elevated haemolymph ammonia on haemocyanin–oxygen affinity of Cancer pagurus

The subtidal crab Cancer pagurus (L.) experiences involuntary periods of emersion associated with practices used in their marketing and distribution. During 24 h emersion, impaired gill function caused an increase of circulating total ammonia (TA=NH₃+NH₄⁺) of 0.35 mmol TA l^-1 (167%). The oxygen-binding characteristics of the haemocyanin of C. pagurus were examined at 10°C in the presence of total ammonia (0.2-1.0 mmol TA l^-1). The haemocyanin-oxygen affinity was decreased in the presence of TA ((logP₅₀/(log[TA]=0.16). Emersion induced significant acidosis and elevated circulating levels of haemolymph TA, lactate and urate, but all had returned to normal levels within 24 h of re-immersion. The accumulation of haemocyanin-modulating substances during 24 h emersion compensated partially (40%) for the effect of the acidosis, but the net effect of the emersion period was a significant decrease in oxygen affinity, corresponding to an increase of P₅₀ (10°C ) from 1.24 kPa (immersed) to 1.96 kPa (24 h emersion). The implications of the findings are considered in terms of the effects and adaptations to emersion.     

Burrowing behaviour and sediment reworking in the heart urchin Brissopsis lyrifera Forbes (Spatangoida)

Sediment reworking due to burrowing and feeding was studied in the spatangoid Brissopsis lyrifera, at two different temperatures (7°C and 13°C). Spine activity and burrowing behaviour were recorded with a real-time video camera. Reworked sediment volume was calculated from tracks produced by the heart urchin. Ingestion rates were measured by feeding the heart urchins with luminophores. Temperature had a significant effect on the bioturbation activity of B. lyrifera. At 13°C reworked sediment volume due to burrowing was 22 ml sediment h^-1 and at 7°C 14 ml sediment h^-1. The ingested amount of sediment was 0.08 and 0.02 g dry sediment h^-1 in 13 and 7°C, respectively. Reworked sediment volume due to burrowing was 60-150 times higher than the volume ingested. The large reworked volume is a consequence of B. lyrifera moving with a rocking motion through the sediment. The spines were continuously transporting sediment around the test with 5-min metachronal wave cycles.     

Temperature adaptation in eurythermal cod (Gadus <Emphasis Type="Italic">morhua</Emphasis>): a comparison of mitochondrial enzyme capacities in boreal and Arctic populations

Activities of citrate synthase (CS), cytochrome c oxidase (CCO) and the electron transport system (ETS) were investigated in white muscle and liver of laboratory-maintained cod, Gadus morhua, from the North Sea, Norwegian coast and Barents Sea for an analysis of temperature acclimation and adaptation in aerobic metabolism. Cold acclimation within each population led to elevated activities of CS, CCO and ETS in white muscle. In liver, however, only North Sea cod showed cold-compensated CS activities, with CCO and ETS unchanged. In contrast, cold-acclimated Norwegian cod displayed unchanged enzyme activities, and Arctic cod showed elevated activities for CS, but decreased activities for CCO and no change in ETS. Between-population comparisons revealed clear evidence for permanent cold adaptation in white muscle of northern (Norwegian coast and Barents Sea) compared to boreal (North Sea) cod populations, reflected by higher activities of CS, CCO and ETS at the same acclimation temperature. Cold-compensated, mass-specific enzyme activities in liver were found in northern compared to boreal cod for CS and CCO, however, for ETS, after warm acclimation only. When evaluated as capacity in total liver, such activities were only found in northern populations in the case of CS at all temperatures, and for CCO and ETS at 15°C only. Hepatosomatic index (I_H) and liver lipid contents were highest in North Sea cod, with similar I_H but lower lipid contents in cold- versus warm-acclimated animals. An acclimation effect on I_H was found in Norwegian cod only, with higher I_H but unchanged lipid contents in the cold. In conclusion, permanent cold adaptation of muscle aerobic metabolism prevails in cod populations at higher latitudes, which is in line with permanently elevated rates of oxygen consumption observed in a parallel study. These differences reflect higher maintenance costs in cold-adapted versus cold-acclimated cod.     

Atmospheric dinitrogen fixation by benthic communities of Tikehau Lagoon (Tuamotu Archipelago, French Polynesia) and its contribution to benthic primary production

Acetylene reduction rates were measured in lagoonal sediments, cyanobacterial mats and limestone surfaces between 1991 and 1995 at many sites, depths and seasons; all the studied substrata contained cyanobacteria. The acetylene reduction/¹⁵N₂ fixation ratio was measured for the different communities and varied between 1.8 and 4.8, depending on substratum. Fixation rates were 1.7 to 7 times higher during daylight compared to night-time rates. N₂ fixation rates ranged from 0.4 to 3.9 mg N m^-2 day^-1 for the lagoonal sediment/mat communities, and the rate was about 2 mg N m^-2 day^-1 for the lagoonal limestone substrata. Total lagoonal benthic N₂ fixation contributed 24.4% of the total nitrogen requirement for the benthic primary production of benthic communities of the lagoon. The input of N₂ fixation by the microbial planktonic communities (including cyanobacteria) of the lagoon, which are highly productive, is unquantified but is likely to be large.     

Life-history traits of Plesionika martia (Decapoda: Caridea) from the eastern-central Mediterranean Sea

Life-history traits of Plesionika martia (Milne Edwards, 1883) were studied through data collected during six seasonal trawl surveys carried out in the Ionian Sea (eastern-central Mediterranean) between July 1997 and September 1998. P. martia was found at between 304 and 676 m depth, with the highest density in the 400-600 m range. Intraspecific, size-related depth segregation was shown. Recruitment occurred in summer at the shallowest depths. Juveniles moved to the deepest grounds as they grew. The largest female and male were 26 and 25 mm carapace length, respectively. The sex ratio was slightly in favour of females at depths >400 m. Although a seasonal spawning peak was shown, the reproduction appears to be rather prolonged throughout the year. Females with ripe gonads were found from spring to autumn. Ovigerous females with eggs in late maturity stage were found year round. Large females could spawn more than one time within their annual reproductive cycle. The size at first maturity (50% of the ovigerous females) was 15.5 mm CL. Average brood size of eggs with a well-developed embryo was 2,966ǃ,521. Iteroparity, low fecundity and large egg size patterns were observed. Brood size increased according to the carapace length. Two main annual groups were found in the field population of the Ionian Sea. Estimates of the Von Bertalanffy growth parameters are: L_X=30.5 mm, k=0.44 year^-1 in females; L_X=28.0 mm, k=0.50 year^-1 in males. A negative allometry was detected mostly in the ovigerous females. The life cycle of P. martia is discussed in the light of life-history adaptations shown in other deep-water shrimp species.