NADP+-isocitrat-dehydrogenase aus Idus idus (Pisces: Cyprinidae). II. Einfluß der temperatur auf substrat- und cosubstrataffinität

Maximum substrate and cosubstrate affinity, as judged by the Michaelis constant (K _M ), of NADP⁺-dependent isocitrate dehydrogenase of pig heart (purchased from Boehringer, Mannheim, FRG) is attained at 37°C. If K _M -values of substrate (Isocitrate, IC) and cosubstrate (NADP⁺) of NADP⁺-dependent isocitrate dehydrogenase (ICDH) of the white dorsal muscle of Idus idus L. is plotted against the experimental temperature (VT), W-shaped curves result. With increasing adaptation temperature (AT), there is a shift to increasing VT. It is suggested that the W-shaped curves are due either to the simultaneous presence of two multiple forms of the enzyme, or to the reversible temperature-dependent interconversion of one protein species.     

NADP+-isocitrat-dehydrogenase aus Idus idus (Pisces: Cyprinidae). I. Aktivität als funktion der adaptationstemperatur

Activity of NADP⁺-dependent isocitrate dehydrogenase (ICDH) of the white dorsal muscle of Idus idus L. was determined by special analysis, under optimum homogenisation, centrifugation and cuvette conditions. The experimental conditions differed considerably from those used for the investigation of NADP⁺-ICDH activity in mammals by Bücher et al. (1964). Daily measurements of activity over a period of 10 days revealed variations in enzyme activity which are difficult to explain. However, the variations are smaller than those established in earlier tests. In order to obtain statistically valid results, the highest possible number of test fishes (at least 10 individuals for each measurement) must be investigated over a period of 5 to 10 days. NADP⁺-ICDH of individuals acclimated to 10°C showed 28% higher specific and non-specific activity than that of fish maintained at 20°C (experimental temperature 25°C). At 5 different adaptation temperatures, increasing activities were observed with decreasing adaptation temperature (compensation). These measurements were made over a period of 5 successive days.     

Metabolism,enzymic activities and cold adaptation in Antarctic mesopelagic fishes

Several species of Antarctic mesopelagic fishes that have different minimal depths of occurrence but the same environmental temperature were collected in November–December 1983 and in March 1986 between 0 and 1 000 m in the open water near the marginal ice zone in the vicinity of 60°S 40°W (1983) and 65°S 46°W (1986), and oxygen consumption rate (V _{O 2}) and the activity of two metabolic enzymes, lactate dehydrogenase (LDH, an indicator of the anaerobic potential of locomotory muscle) and citrate synthase (CS, an indicator of citric acid cycle activity or aerobic potential), were determined. In four dominant species, whole-individual oxygen-consumption rate (y, ml O₂ individual^–1 h^–1) varied with weight (X, g) according to the equation y=aX ^b, with b values falling between 0.889 and 1.029. The relation of weight-specific LDH activity (y, U g^–1 wet wt) with weight (x, g) was also described by the equation y=aX ^b, with b values varying between 0.229 and 1.025. Weight-specific CS activity declined with weight, with b values from-0.031 to-0.369. V _{2 O}, LDH activity and CS activity all declined markedly with increased species' minimum depth of occurrence (the depth below which 90% of a species' population lives). Comparisons with previous studies on ecologically equivalent species of the California Borderland indicate that depth-related decreases in metabolism are the result of adapted traits of deeper-living species, not declining temperature within the water column. The metabolic rate of Antarctic mesopelagic fishes is approximately twice that of California species at equivalent temperatures; similar rates were found at the normal habitat temperatures of the two groups. Thus, a well-developed compensation for temperature is present in the Antarctic fishes: cold adaptation. Differences in enzymic activity among species, and among different sized individuals of a species are related to differences in metabolic rate and locomotory capacity. Enzymic indices can be used to estimate metabolic rates and evaluate ecological parameters such as predatory strategies and niche separation.     

Der einfluß der temperatur auf thermostabilität,isoenzym-muster und reaktionskinetik der laktat-dehydrogenase aus fischen

Methodological problems complicate investigations on thermostability of lactate dehydrogenase (LDH). It is difficult to demonstrate a correlation between adaptation-temperature (AT) and LDH thermostability. Heat-inactivation characteristics change completely if diluted or undiluted tissue extracts are heated. In purified LDH (purchased from Boehringer, Mannheim, FRG), additions such as casein, bovine-serum albumin, NADH and pyruvate — even in small concentrations — can alter considerably the degree of heat resistance. It LDH activity is measured as a function of experimental temperature (ET) according to the composition of the actual test mixture (e.g. altered pyruvate concentration), a different temperature optimum is found. If tissue extracts containing unpurified enzymes are used, the accompanying substances act on the enzyme and modify its properties. Thus, possible influence of AT on enzyme properties can be concealed (suppressed, over-emphasized). In Idus idus acclimated to 10° or 20°C, brain, gill, gut and white dorsal muscle reveal identical LDH-isoenzyme patterns. However, liver-LDH shows a pattern dependent on the AT. A total of 11 bands with LDH activity were found. In 10°C fishes, the Isoenzymes 1, 3, 6 and 7 are especially active. However, 20°C fishes show marked activity of Isoenzymes 5 and 8, and a reduced activity of Isoenzyme 7. According to their electrophoretic mobility, the particular isoenzymes of LDH of white dorsal muscle of I. idus or Rhodeus amarus can be clearly distinguished. The ATs 10° or 20°C do not influence the dependence of reaction order on ET: this is not true for the velocity constant.     

Effects of nutrition and temperature on metabolic enzyme activities in larval and juvenile red drum,Sciaenops ocellatus,and lane snapper,Lutjanus synagris

The metabolic enzyme activities were determined in larvae of red drum, Sciaenops ocellatus, and lane snapper, Lutjanus synagris, to determine the effect of temperature and nutrition on metabolic enzyme activities and to evaluate if metabolic enzyme activities are useful in assessing the feeding condition of larval fish. During experiments conducted during the spring of 1990, lactate dehydrogenase (LDH) activities in both red drum and lane snapper were approximately an order of magnitude lower than values typical for adult fish; LDH and citrate synthase (CS) activities increased during early developmental stages, but nutritional effects were apparent. Clear differences (up to 4-fold) between well-fed and starving fish were evident in both LDH and CS activity in red drum. Differences between well-fed and poorly fed larvae were evident until 9 d after hatching. Lane snapper larvae reared at a 25°C had significantly lower LDH activities than larvae reared at 28°C.     

L-Lactate dehydrogenase in the longitudinal muscle of the sea cucumber Sclerodactyla briareus (Echinodermata: Holothuroidea)

The infaunal holothurian Sclerodactyla briareus (Thyone briareus) is able to tolerate exposure to hypoxic conditions for over 2 days. Since the in vitro anaerobic degradation of glucose-U-C¹⁴ by longitudinal muscle preparations leads to an accumulation of labeled lactic acid, it is apparent that lactate dehydrogenase plays a key role during anoxia. Disc electrophoresis resolved one major band of lactate dehydrogenase (LDH) activity in longitudinal muscle extracts. Muscle extracts of s. briareus oxidized L-lactate only, as is the case with most other echinoderms. The apparent Michaelis constants for lactate oxidation and pyruvate reduction are dissimilar from values reported for other marine invertebrates. The LDH activity in this holothurian is higher than the activities reported for other marine invertebrates with the exception of some active, large arthropods. Substrate inhibition of LDH maximal activity occurs at substrate concentrations (pyruvate) exceeding 10 mM, and then to only a minor extent. The characteristics of lactate dehydrogenase from the longitudinal muscle of S. briareus support the hypothesis that it functions to maintain glycolytic flux during prolonged periods of exposure to anoxic or hypoxic conditions.     

Depth zonation and metabolic adaptation in Dover sole,Microstomus pacificus,and other deep-living flatfishes: factors that affect the sole

Flatfishes of Monterey Bay, central California, undergo species replacements with increasing depth along a transect from 100 m on the continental shelf down to a depth of 1400 m on the continental slope. The Dover sole, Microstomus pacificus, differs from the other local flatfish species by undergoing an extensive ontogenetic vertical migration, occupying all depth zones at different life stages, and having its maximum spawning biomass in the oxygen minimum zone between 600 and 1000 m. Size-activity relationships and depth-activity relationships for the glycolytic enzyme lactate dehydrogenase (LDH) and for two enzymes associated with aerobic metabolism, malate dehydrogenase and citrate synthase (CS), were examined in white-muscle tissue of shallow-living, deep-living and ontogenetically-migrating species. Scaling coefficients (b) for weight-specific enzyme activity (log activity)=a+b (log wet weight), varied in sign as well as magnitude for fishes living at different depths. In the shallow-living California halibut Paralichthys californicus, LDH scaled positively (0.39) and CS scaled negatively (-0.15) with size, a pattern observed previously for most shallow-water fish species. The permanently deep-living species, the deepsea sole Embassichthys bathybius, differed in that both LDH and CS scaled strongly negative (-2.0 and-1.5, respectively). For the ontogenetically migrating Dover sole Microstomus pacificus, there was a shelf-slope transition. For the shelf specimens (200 m), LDH scaled positive (0.11) and CS negative (-0.29) and for the slope specimens (400 m), LDH scaled negative (-0.65) and CS strongly negative (-0.63). Rex sole, Glyptocephalus zachirus, showed a similar shelf-slope transition. Intraspecific depth-enzyme activity differences were not incremental, but changed abruptly between the continental shelf stations (100 to 200 m) and the continental slope (400 to 1400 m). Based on comparisons with laboratory-maintained individuals, the decline in the metabolic capacity of the white muscle of Dover sole is a phenotypic response to the low food and oxygen conditions of the continental slope. Contrary to expectation, anaerobic capacity (LDH activity) decreased in response to low oxygen conditions, suggesting that in a permanently hypoxic environment such as the oxygen minimum zone the metabolic strategy may be to not incur an oxygen debt that would be difficult to pay back.     

Glucose-6-phosphate dehydrogenase and thermal acclimation in the mullet fish

Glucose-6-phosphate dehydrogenase in mullet liver (Mugil cephalus L.) occurs in 2 electrophoretically distinguishable forms, termed I and II. Both are tetramers (at neutral pH) with molecular weights greater than 200,000. At pH 9, each tetramer dissociates into two dimer molecules which retain catalytic activity. At intermediate pH values, an equilibrium tetramer dimer mixture probably occurs. Kinetic studies of Form I indicate that it is strongly inhibited by TPNH, which is competitive with respect to both G6P and TPN. Inhibition is greatly increased at high temperatures while enzyme-substrate and enzymeecoenzyme affinities greatly decrease. These properties strongly favor glucose-6-phosphate dehydrogenase function at low temperature, a situation that is partly compensated for during warm acclimation by an increase in the relative importance of Form II. Although Form II displays catalytic and regulatory properties which are qualitatively similar to those of Form I, it is able to outcompete Form I for limiting substrate at high temperatures.     

Purification and some properties of β-N-acetyl-D-glucosaminidase from prawn (<Emphasis Type="Italic">Penaeus vannamei</Emphasis>)

The -N-acetyl-D-glucosaminidase (NAGase, EC 3.2.1.52) from prawn (Penaeus vannamei) was purified by extraction with 30% ethanol solution and ammonium sulfate fractionation, then chromatographed on Sephadex G-100 followed by DEAE-cellulose (DE-32) columns. The purified enzyme determined to be homogeneous by polyacrylamide gel electrophoresis (PAGE) and SDS-PAGE. The specific activity of the purified enzyme was 1,560 U mg^–1. Enzyme molecular weight was determined to be 105,000 Da; it contained two subunits of the same mass (45,000 Da). The pI value was calculated to be 4.8 by isoelectric focusing. The optimum pH and optimum temperature of the enzyme for the hydrolysis of pNP--D-GlcNAc (enzyme substrate) were determined to be pH 5.2 and 45°C, respectively. The behavior of the enzyme during hydrolysis of pNP--D-GlcNAc followed Michaelis–Menten kinetics, with K_m=0.254 mM and V_m=9.438 M min^–1, at pH 5.2 and 37°C. The stability of the enzyme was investigated, and the results showed that the enzyme was stable in a pH range from 4.2 to 10.0 and at temperatures <40°C. The effects of metal ions on the enzyme were also studied. Li⁺, Na⁺ and K⁺ had no influence on enzyme activity. Mg²⁺, Ca²⁺ and Mn²⁺ activated the enzyme, while Ba²⁺, Zn²⁺, Co²⁺, Cd²⁺, Hg²⁺, Pb²⁺ Cu²⁺, Fe³⁺ and Al³⁺ showed various degrees of inhibitory effects on the enzyme.Communicated by O. Kinne, Oldendorf/Luhe     

Alterations in plasma electrolyte levels of a freshwater fish Cyprinus carpio exposed to acidic pH

The impact of acidification (Low pH 5.0) on plasma electrolytes (Na⁺, K⁺, Cl^?, Ca²⁺, and Mg²⁺) in a freshwater fish Cyprinus carpio was studied for a 35 day (long term) exposure period, while control groups were maintained at neutral pH (7.3). During long-term (35 days) exposure periods, plasma K⁺ and Mg²⁺ levels were increased (77.8% and 16.0%) in the low pH (5.0) treated fish. On the other hand, plasma Na⁺, Cl^?, and Ca²⁺ levels were decreased (12.4%, 18.4% and 31.3%, respectively). The loss of plasma Na⁺, Cl^?, and Ca²⁺ indicates the displacement of Ca²⁺ from tight junctions. The increased plasma K⁺ ion might have resulted from acidosis, because intracellular K⁺ is released from muscle as H⁺ enters. The elevated level of plasma Mg²⁺ might be due to inhibition of active transport of magnesium across the kidneys resulting in the accumulation of this ion in the plasma. Ionic alteration takes place upon exposure to acidic pH and can be considered as a potential tool for detecting environmental stresses caused by acidification.     

Gas–liquid chromatography for fenvalerate residue analysis: alterations in the ionic concentration and associated adinosine triphosphatases in fish,Cirrhinus mrigala (hamilton)

In the present study, an attempt has been made to quantify the fenvalerate accumulated in different tissues (gill, muscle and liver) and observe changes involved in the levels of sodium, potassium and calcium ions and Na⁺–K⁺, Mg²⁺ and Ca²⁺ adenosine triphosphatase (ATPase) activities in the freshwater fish, Cirrhinus mrigala on short-term and long-term exposure to the median lethal and sublethal concentration of fenvalerate. Residue analysis using gas–liquid chromatography (GLC) technique revealed that fenvalerate accumulated in highest quantity in gill followed by liver and muscle under median lethal concentration (6?µg?L^?1). Whereas in sublethal concentration (0.6?µg?L^?1), muscle accumulated highest quantity followed by gill and liver, which might be due to the fact that fenvalerate is highly lyphophilic. The ion concentration and ATPase activity were found effected in fish exposed to lethal and sublethal concentrations of fenvalerate. Concentration of Na⁺, K⁺ and Ca²⁺ ions decreased in gill, muscle and liver on being exposed to median lethal concentration to a significant level. Whereas the changes were not highly pronounced at sub lethal level indicating low concentration of fenvalerate and its non-toxic effect at chronic exposure. Na⁺–K⁺, Mg²⁺ and Ca²⁺ ATPases activity were also found decreased in correspondence to the ionic change under median lethal and sub lethal concentrations in target tissues. This might have lead to behavioural changes and create wide-spread disturbance in the normal physiology, ultimately causing the death of the fish. The results suggest that in biomonitoring programmes, ions and associated ATPases can be a good diagnostic tool for fenvalerate toxicity.     

Effect of salinity fluctuation on the osmotic pressure and Na+, Ca2+ and Mg2+ ion concentrations in the hemolymph of bivalve molluscs

Specimens of Chlamys opercularis, Modiolus modiolus, Mytilus edulis, Crassostrea gigas, Scrobicularia plana and Mya arenaria were exposed to both gradual (sinusoidal) and abrupt (square-wave) salinity fluctuations and measurements made of osmotic, Na⁺, Mg²⁺ and Ca²⁺ concentrations in the hemolymph and where applicable in the mantle fluid. In both sinusoidal and square-wave regimes fluctuating between 100 and 50% seawater (100%=ca. 32 S), the hemolymph Na⁺, Mg²⁺, Ca²⁺ and osmotic concentrations followed the concentrations of the external medium in Chlamys opercularis. The hemolymph and mantle fluid osmotic Na⁺, Mg²⁺ and Ca²⁺ concentrations of Modiolus modiolus, Mytilus edulis, Crassostrea gigas and S. plana followed those of the external medium as long as the molluscs' shell valves remained open. There were no changes in the ionic or osmotic concentrations of the hemolymph or mantle fluid of any of these species during periods of shell-valve closure. The hemolymph osmotic, Na⁺ and Mg²⁺ concentrations of wedged-open Modiolus modiolus, Mytilus edulis, C. gigas and S. plana followed those of the external medium. Hemolymph Ca²⁺ concentrations showed a damped response in C. gigas and Mytilus edulis. The hemolymph osmotic, Na⁺, Ca²⁺ and Mg²⁺ concentrations of Mya arenaria fluctuated in a similar manner to the external medium, but were damped. Wedged-open Mytilus edulis exposed to fluctuating salinity and supplied with a constant supply of 10 mM Ca²⁺ showed greater changes in hemolymph ionic and osmotic concentrations than M. edulis exposed to the same salinity fluctuation without a constant Ca²⁺ supply. Chlamys opercularis and Modiolus modiolus survived in a 50% seawater minimum sinusoidal salinity fluctuation for 10 days; wedged-open M. modiolus survived only 3 days. Burrowing had no effect on the osmotic, Na⁺, Mg²⁺ or Ca²⁺ concentrations of the hemolymph of Mya arenaria or S. plana exposed to fluctuating salinities. All of the species studied were shown to be osmoconformers.     

ZnO纳米粒子通过线粒体通路抑制小鼠光感受器细胞Na~+/K~+-ATP酶表达和活性的作用研究

氧化锌(ZnO)纳米粒子已被发现具有生物毒性,氧化应激被认为是最重要的因素之一。前期实验证实,ZnO纳米粒子能显著减少锰超氧化物歧化酶(Mn SOD)蛋白的表达,降低Mn SOD活性。本文通过检测乳酸脱氢酶(LDH)释放、线粒体活性氧(ROS)水平和膜电位(Δφm)、延迟整流钾电流变化和Na~+/K~+-ATP酶的表达及活性等变化,检测ZnO纳米粒子对小鼠光感受器细胞的细胞毒作用。结果表明,ZnO纳米粒子可显著增强小鼠光感受器细胞中LDH的释放、增加线粒体内ROS水平并下调Δφm、阻断延迟整流钾电流,同时降低Na~+/K~+-ATP酶的表达及活性,从而对小鼠视网膜光感受器细胞产生细胞毒作用,提示ZnO纳米粒子可通过线粒体通路引起氧化应激,从而抑制小鼠光感受器细胞Na~+/K~+-ATP酶表达和活性,产生细胞毒性,导致细胞死亡。本文的研究结果有助于理解ZnO纳米粒子引起细胞毒性的作用机理。     

Physiological responses to fluctuation in temperature of salinity in invertebrates. Adaptations ofAlpheus viridari (Decapoda,Crustacea),Terebellides parva (Polychaeta) andGolfinigia cylindrata (Sipunculida) to the mangrove habitat

The snapping shrimpAlpheus viridari (Armstrong, 1949), the polychaeteTerebellides parva Solis-Weiss, Fauchald and Blankensteyn 1990, and the sipunculanGolfingia cylindrata (Keferstein, 1865) are commonly found in the same mangrove habitat, where they experience frequent, acute fluctuations in temperature and salinity. Ecological studies indicate a temporal variation, including occasional absence, in the distribution of bothG. cylindrata andT. parva; this fed us to examine the physiological adaptations of the three species (collected at Western Bay, Twin Cays, Belize in 1985, 1986 and 1988). Each was subjected to acute, repeated exposure to either control (35 S) and decreased (25 S) salinity or to control and increased (45 S) salinity. Ability to regulate water and ion content (g H₂O or mol g^-1 solute free dry wt) was examinedA. viridari behaved as a hyperosmotic conformer at decreased salinity but as an osmoconformer at increased salinity. Regardless of direction of salinity change,A. viridari regulated water content through change in Na⁺, K⁺, and Cl^– contents. In contrast,G. cylindrata behaved as an osmoconformer and did not demonstrate ability to regulate water content.T. parva behaved as an osmoconformer, showed incomplete regulation of water content via change in Na⁺, K⁺, and Cl^– contents but had limited survival following exposure to 45 S. Each species was also exposed to change in temperature. Species were subjected to acute, repeated exposure either to control (28°C) and decreased (21°C) temperature or to control and inereased (35°C) temperatureA. viridari regulated water and ion content under both experimental conditions. In contrast,T. parva did not regulate water and ion content under either experimental temperature.G. cylindrata did not regulate water and ion content during exposure to decreased temperature and did not survive exposure to increased temperature. ForA. viridari, weight specific oxygen uptake rates (mg O₂ g^-1 ash-free dry wt) were determined. Exposure to decreased salinity or to increased temperature resulted in a small sustained elevation in O₂ uptake. It is concluded that, unlikeA. viridari, T. parva andG. cylindrata are only marginally adapted to withstand the salinity and temperature stresses, respectively, of the mangrove habitat. The inability ofT. parva andG. cylindrata to fully adapt to extremes in the mangrove habitat could well explain the temporal variation seen in the distribution of these two species.Contribution number 380. Caribbean Coral Reef Ecosystems, National Museum of Natural History, Smithsonian Institution     

The influence of suspension density and temperature on the filtration rate of Hiatella arctica

The rate of filtering Phaeodactylum tricornutum and Isochrysis galbana was measured in Hiatella arctica (L.) by the indirect suspension depletion method monitored by optical density measurement. The filtration rate of H. arctica was found to be 1.412×10^–2 l/h/g wet weight at a temperature of 15°C when fed with P. tricornutum, at average cell concentrations up to 3.5×10⁶ cells/ml. The filtration rate dropped almost to zero when the concentration of P. tricornutum reached 11×10⁶ cells/ml. The filtration rate of I. galbana diminished at a much lower cell concentration of 1×10⁶ cells/ml, and almost ceased at 3 to 4×10⁶ cells/ml. In mixed cultures of I. galbana and P. tricornutum, the filtration rate ratio was 0.37 to 1.00, and this was believed to be due to a proportion of the smaller former cells passing through the ostia. However, when resuspended in sea water, I. galbana cells were taken at a rate slightly less than P. tricornutum. The medium in which the I. galbana cells had been grown was inhibitory to the filtering activity of H. arctica, since, when cells of either alga were resuspended in the medium, the filtration rate was considerably reduced. No inhibitory factor existed in either of the original nutrient media. Hence, the importance of using low cell concentrations and of eliminating any inhibitory metabolic products when measuring filtration rates of bivalves is stressed. H. arctica shows a typical activity temperature eurve for a boreo-arctic species, with a steady rise from 0°C to a maximum between 15° and 17°C, and a sharp fall in activity to about zero at 25°C. The rates of filtration of various species at temperatures approaching the optimum were compared after allowance was made for fall in filtration rate with increasing body weight. The results suggested that the Mytilacea had the highest filtration rates and that H. arctica possesses one of the lowest filtration rates recorded.     

Über den Einfluß der Nahrungskonzentration und anderer Faktoren auf Filtrierleistung und Nahrungsausnutzung der Muscheln Arctica islandica und Modiolus modiolus

Filtration rates and the extent of phagocytosed food particles were determined in the offshore lamellibranchs Artica islandica and Modiolus modiolus in relation to particle concentration, body size and temperature. Pure cultures of the algae Chlamydomonas sp. and Dunaliella sp. were used as food. A new method for determining filtration rates was developed by modifying the classical indirect method. The concentration of the experimental medium (100%) was kept constant to ±1%. Whenever the bivalves removed algae from the medium, additional algae were added and the filtration rate of the bivalves expressed in terms of percentage amount of algae added per unit time. The concentration of the experimental medium was measured continuously by a flow colorimeter. By keeping the concentration constant, filtration rates could be determined even in relation to different definite concentrations and over long periods of time. The amount of phagocytosed food was measured by employing the biuret-method (algae cells ingested minus algae cells in faeces). Filtration rates vary continuously. As a rule, however, during a period of 24 h, two phases of high food consumption alternate with two phases of low food consumption during which the mussels' activities are almost exclusively occupied by food digestion. Filtration rate and amount of phagocytosed algae increase with increasing body size. Specimens of A. islandica with a body length of 33 to 83 mm filter between 0.7 to 71/h (30–280 mg dry weight of algae/24 h) and phagocytose 21 to 122 mg dry weight of algae during a period of 24 h. The extent of food utilization declines from 75 to 43% with increasing body size. In M. modiolus of 40 to 88 mm body length, the corresponding values of filtration rate and amount of phagocytosed algae range between 0.5 and 2.5 l/h (20–100 mg dry weight of algae) and 17 to 90 mg dry weight of algae, respectively; the percentage of food utilization does not vary much and lies near 87%. Filtration rate and amount of phagocytosed algae follow the allometric equation y=a·x ^b. In this equation, y represents the filtration rate (or the amount of phagocytosed algae), a the specific capacity of a mussel of 1 g soft parts (wet weight), x the wet weight of the bivalves' soft parts, and b the specific form of relationship between body size and filtration rate (or the amount of phagocytosed algae). The values obtained for b lie within a range which indicates that the filtration rate (or the amount of phagocytosed algae) is sometimes more or less proportional to body surface area, sometimes to body weight. Temperature coefficients for the filtration rate are in Arctica islandica Q₁₀ (4°–14°C)=2.05 and Q₁₀ (10°–20°C)=1.23, in Modiolus modiolus Q₁₀ (4°–14°C)=2.33 and Q₁₀ (10°–20°C)=1.63. In A. islandica, temperature coefficients for the amount of phagocytosed algae amount to Q₁₀ (4°–14°C)=2.15 and Q₁₀ (10°–20°C)=1.55, in M. modiolus to Q₁₀ (4°–14°C)=2.54 and Q₁₀ (10°–20°C)=1.92. Upon a temperature decrease from 12° to 4°C, filtration rate and amount of phagocytosed algae are reduced to 50%. At the increasing concentrations of 10×10⁶, 20×10⁶ and 40×10⁶ cells of Chlamydomonas/l offered, filtration rates of both mollusc species decrease at the ratios 3:2:1. At 12°C, pseudofaeces production occurs in both species in a suspension of 40×10⁶, at 20°C in 60×10⁶ cells of Chlamydomonas/l. At 12°C and 10–20×10⁶ cells of Chlamydomonas/l, the maximum amount of algae is phagocytosed. At 40×10⁶ cells/l, the amount of phagocytosed cells is reduced by 26% as a consequence of low filtration rates and intensive production of pseudofaeces. At 20°C and 20–50×10⁶ cells of Chlamydomonas/l, the maximum amount of algae is sieved out and phagocytosed; the concentration of 10×10⁶ cells/l is too low and cannot be compensated for by increased activity of the molluscs. With increasing temperatures, the amount of suspended matter, allowing higher rates of filtration and food utilization, shifts toward higher particle concentrations; but at each temperature a threshold exists, above which increase in particle density is not followed by increase in the amount of particles ingested. Based on theoretical considerations and facts known from literature, 7 different levels of food concentration are distinguishable. Experiments with Chlamydomonas sp. and Dunaliella sp. used as food, reveal the combined influence of particle concentration and particle size on filtration rate. Supplementary experiments with Mytilus edulis resulted in filtration rates similar to those obtained for M. modiolus, whereas, experiments with Cardium edule, Mya arenaria, Mya truncata and Venerupis pullastra revealed low filtration rates. These species, inhabiting waters with high seston contents, seem to be adapted to higher food concentrations, and unable to compensate for low concentrations by higher filtration activities. Adaptation to higher food concentrations makes it possible to ingest large amounts of particles even at low filtration rates. Suspension feeding bivalves are subdivided into four groups on the basis of their different food filtration behaviour.     

Ammonia production and kinetic properties of glutamate dehydrogenase in the sipinculid Phascolosoma arcuatum exposed to anoxia

The amounts of total NH ₄ ⁺ detected in the external media in which Phascolosoma arcuatum had been exposed to various periods of anoxia were significantly greater than those in which the worms were exposed to normoxia for a similar period. The increased NH ₄ ⁺ production by P. arcuatum during anoxic exposure was unlikely to be due to an increased catabolism of adenine nucleotides or urea. In contrast, there were significant decreases in the concentrations of several free amino acids in the coelomic plasma and body tissues of individuals during the 48 h of anoxic exposure. The amount of NH ₄ ⁺ produced by the anoxic P. arcuatum could be accounted for by the decreases in the concentrations of aspartate or glycine. Increases in the catabolism of free amino acids (FAA), leading to the increased production of NH ₄ ⁺ , in P. arcuatum during anoxia were supported by the detection of significant changes in the kinetic properties of glutamate dehydrogenase (GDH), in the deaminating direction, from worms exposed to anoxia for 48 h. The apparent increase in the affinity of GDH from the anoxic worm to glutamate would bring about a greater deaminating activity at physiological concentrations of ths substrate. P. arcuatum used in these experiments were collected from the mangrove swamp at Mandai, Singapore between 1990 and 1993.     

Interacting pressure and temperature effects on enzymes of marine poikilotherms: Catalytic and regulatory properties of FDPase from deep and shallow-water fishes

At low temperature (3°C), and in the absence of substrate and cofactor, the enzyme, fructose diphosphatase (FDPase), extracted from liver of the benthic fishCoryphaenoides acrolepis, is reversibly inactivated by exposure to relatively low pressures (5,000 psi). At pressures of 20,000 psi at 3°C the native enzyme is irreversibly denatured. When the cofactor and the substrate are present, the enzyme is protected against pressure denaturation; hence, catalysis is insensitive to at least 10,000 psi at 3°C. Similarly, interactions between FDPase and its negative modulator adenosine monophosphate (AMP), are largely pressure insensitive. Pressure sensitivity of the native enzyme and of the catalytic process is much reduced at temperatures above about 9°C. The homologous enzyme from a surface fish,Pimelometopon pulchrum, is strikingly more sensitive to pressure, particularly at low temperatures.     

Grazing of Acartia hudsonica (A. clausi) on Skeletonema costatum in Narragansett Bay (USA): Influence of food concentration and temperature

Grazing experiments were performed with temperatureacclimated Acartia hudsonica fed the diatom Skeletonema costatum in concentrations ranging from 50 to 3×10⁴ cell ml^-1 at 5°, 10° and 15°C. The ingestion data were best fit by an Ivlev equation. Feeding threshold values of 39 and 59 cells ml^-1 were not significantly different from zero; however, filtration rates were depressed at low food concentrations. Maximum filtration rates increased exponentially with temperature, reaching a maximum with copepods collected at 14°–15°C, and then declining. Both the increase in ingestion rate with increasing food concentration and the maximum ingestion rate were significantly greater as experimental temperature was increased. Maximum ingestion rates were reached at concentrations greater than 6×10³ cells ml^-1. Percent of body carbon ingested per day at 5 g C L^-1 increased from 1.5% at 5°C to 6.7% at 15°C. At 500 g C L^-1, the ingestion increased from 84% (5°C) to 660% (15°C). Percent of body nitrogen at 0.5 g N L^-1 increased from 0.6% per day at 5°C to 2.5% per day at 15°C. At 50 g N L^-1, the ingestion was 42% body nitrogen at 5°C and 250% at 15°C. The influence of grazing by A. hudsonica on phytoplankton in Narragansett Bay, USA was estimated for 1972–1977. The percent of standing stock removed by grazing rarely exceeded 5% per day except during the late spring when S. costatum growth becomes nutrient limited and higher temperatures favor the rapid population growth of A. hudsonica.     

The effect of pressure on muscle lactate dehydrogenase activity of some deep-sea and shallow-water fishes

The activity of crude muscle lactate dehydrogenase (LDH) of several species of bathypelagic and shallow-water fishes has been measured at pressures between 1 and 578 atm and at temperatures of 15° and 25°C. No relationship has been found between the effect of pressure on enzyme activity and the hydrostatic pressure of the organism's environment. Applied hydrostatic pressure reduced activity at both temperatures. The decrease at 25°C was double the decrease at 15°C in LDH from shallow-water fishes. However, enzymes from 2 bathypelagic fishes showed approximately the same reduction at both temperatures. Thus, the interaction of temperature and pressure was less in deep-sea than in shallow-water fish LDH. Decreasing temperature and increasing pressure would both reduce the activity of LDH. That is, deep-sea conditions are noncompensatory in this instance. It is possible that the dissociation of the effects of temperature and pressure could be an adaptive feature of deep-sea life.This paper is a portion of a thesis submitted to the Graduate School, University of Georgia, in partial fulfillment for the degree of Master of Science.