Temperature acclimation in respiratory and cytochrome c oxidase activity in common carp (Cyprinus carpio)

• 1.1. Common carp (Cyprinus carpio) exposed to experimental temperatures of 12, 18, 24, 30 or 36°C for a 4-week period were used to investigate the effect of temperature acclimation on the frequency of opercular movement (FOM), growth and cytochrome c oxidase (CCO) activity in heart, liver and muscle.
• 2.2. An exponential relationship between FOM and temperature after the first week (10₁₀ =1.76) disappeared after the second week.
• 3.3. The initially high FOM at temperatures of 30 or 36°C and the low FOM at 18 or 12°C changed over 4 weeks to approach the FOM of fish at 24°C.
• 4.4. This change in the relationship of FOM to temperature from highly dependent to independent appeared to be thermal compensation.
• 5.5. Heart and liver CCO activities were significantly affected by temperature, with the lowest activity at the approximate optimum temperature for growth, 24°C.
• 6.6. Highest CCO activities for heart and liver occurred at both the highest and lowest temperatures.
• 7.7. Among the three tissues, heart CCO activity was generally the highest and most affected by acclimation temperature.
• 8.8. Muscle tissue had the lowest CCO activity and was unaffected by temperature.
• 9.9. The high CCO activity at a cold acclimation of temperature 12°C was probably due to thermal compensation and the high activity at 36°C may have been a result of thermal stress.

     

Digestive protease,lipase and amylase activities in stage i larvae of the american lobster,Homarus americanus

• 1.1. Digestive protease, lipase, and amylase of Stage I larvae of the American lobster Homarus americanus are characterized.
• 2.2. A sensitive method for detection of crustacean lipase was developed using an latroscan which combines thin-layer chromatography and flame ionization detection to quantify free fatty acids generated by lipase digestion.
• 3.3. pH optima of the three enzymes occurred at or near the pH of gastric fluid.
• 4.4. A time course study demonstrated slight increases in protease and amylase activities during the first larval stage, regardless of whether the lobsters were fed or not, whereas lipase activity was constant.

     

Protease,amylase and lipase activities in the midgut and hindgut of the cricket,Gryllus rubens and mole cricket,Scapteriscus acletus

• 1.1. Midgut is the major source of protease, amylase and lipase in a cricket, Gryllus rubens and in a mole cricket, Scapteriscus actetus.
• 2.2. Hindgut makes a significant contribution, and possibly even the major contribution, to digestion in both crickets, with enzyme activities from 20% (amylase and lipase) to 30% (protease) of midgut level, and a pH favorable to action of all three.
• 3.3. Ingested food helps regulate digestive enzyme levels, and crickets starved for 5 days had only 50–60% of normal levels of enzyme activity.

     

The digestive enzymes of the pacific brown shrimp Penaeus californiensis.: I—Properties of amylase activity in the digestive tract

• 1.1. Crude extract of the whole digestive tract from the brown shrimp (P. californiensis) was investigated for digestive amylase activity.
• 2.2. Considerable amylase activity was found at pH 6.5–8.0, with optimum pH at around 7.5.
• 3.3. Optimum temperature was found between 30–40°C, similar to amylases from other crustaceans.
• 4.4. Amylase activity was highly halotolerant, having 50% maximum activity at 3 M NaCl.
• 5.5. Maximum amylase activity was found at 0.01 M NaCl.
• 6.6. Amylase activity was partially inhibited by the divalent ions Hg²⁺, Zn²⁺, Cu²⁺ and Cr²⁺.
• 7.7. Mg²⁺ and Ca²⁺ ions seemed to enhance amylase activity.

     

Cardiac frequency compensation responses of adult blue crabs (Callinectes sapidus Rathbun) exposed to moderate temperature increases

• 1.1. Cardiac frequency patterns of Callincctes sapidus Rathbun were used to evaluate potential thermal stress after exposure to 5°C increases over a range of acclimation temperatures from 5° to 30°C.
• 2.2. An acclimated rate-temperature curve (R-T curve), acute R-T curves of the stabilized rates at the increased temperatures and Q₁₀ temperature coefficients were used to assess the significance of the changes in rate frequency.
• 3.3. The acclimated R-T curve showed that blue crabs go through a series of seasonal adaptation types characterized by a plateau of perfect adaptation for both cold and warm adapted organisms. Paradoxical adaptation occurred between the transition from cold to warm acclimation temperatures.
• 4.4. The acute R-T curves showed that cardiac frequency was highly responsive to a 5°C increase when the organisms were acclimated to low temperatures.
• 5.5. The Q₁₀'s of the acute R-T curves at the warm acclimation temperatures approximated those values derived for the acclimated R-T curve.
• 6.6. This suggests that the temperature increase had a negligible effect on the warm adapted crabs, that is, little or no thermal stress occurred.

     

Characterization of NaK ATPase from the gills of the freshwater prawn Macrobrachium rosenbergii (de Man)

• 1.1. The specific activity of Na-K ATPase was determined from the microsomal preparation of gills dissected from adult Macrobrachium rosenbergii.
• 2.2. Maximal ATPase activity was achieved at a substrate concentration of 0.5 mM ATP.
• 3.3. Optimal enzyme activity was obtained at pH of 7.5.
• 4.4. The Arrhenius plot of Na-K ATPase activity revealed a marked discontinuity at 30°C. “Mg” ATPase activity did not exhibit a marked discontinuity.
• 5.5. The E_a for Na-K ATPase and “Mg” ATPase was 14.6 kCal/mole and 9.31 kCal/mole respectively. Q₁₀ values for Na-K ATPase was 2.34 and for “Mg” ATPase 1.65.
• 6.6. ATPase activity and gill homogenate protein concentration exhibited a linear relationship up to 130 μg protein/ml.
• 7.7. Na-K ATPase activity was inhibited by 10⁻³ M ouabain. It was equally inhibited by the removal of K⁺ from the reaction medium.

     

Biomines-stimulated phosphorylation and (Na+, K+-ATPase in the gills of the chinese crab,Eriocheir sinensis

• 1.1. Subcellular distribution of (NA⁺, K⁺-ATPase and ouabain-insensitive ATPase (Mg²⁺-ATPase) are compared in branchial tissues of the euryhaline crab, Eriocheir sinensis, acclimated to fresh water.
• 2.2. Both the anterior and posterior gills contain cAMP-dependent protein kinase and endogenous protein substrate for phosphorylation.
• 3.3. Phosphorylation occurs in both “particulate” and “soluble” subcellular fractions but its stimulation by cAMP is restricted to the “soluble” fraction.
• 4.4. serotonin (5-HT) and dopamine receptors are present only in the “light particulate” fraction isolated from the posterior gills.

• 1.(a) Serotonin and dopamine have no effect on the phosphorylation observed in a subcellular fraction alone.
• 2.(b) Activation of the phosphorylation by serotonin and dopamine is found when the soluble fraction (source of cAMP-dependent protein kinase) is added to the fraction P₃ from the posterior gills.
• 3.(c) No activation occurs with the fractions P₃ as well as P₁ or P₂ (not shown) from anterior gills of fresh water crab.
• 4.(d) Cyproheptadine, a serotonin receptor antagonist, inhibits the 5-HT dependent increase in phosphorylation.
• 5.(e) The dopamine receptor antagonist, chlorpromazine, inhibits dopamine-stimulated phosphorylation.
• 6.5. Ouabain mimics the effect of cyproheptadine on the serotonin-stimulated phosphorylation found in the posterior gills.

     

Alkaline phosphatase from Basidiobolus haptosporosus: Comparison with alkaline phosphatases from rainbow lizard and man

• 1.1. DEAE-cellulose chromatography of mycelial alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1) from Basidiobolus haptosporosus, produced three iso-enzymes “A”, “B” and “C”.
• 2.2. Fraction “A” was further characterized and showed maximum activity at pH 10 in 0.1 M sodium carbonate-bicarbonate buffer.
• 3.3. The enzyme was stimulated by Mg²⁺, Co²⁺ and Mn²⁺ and inactivated by Zn²⁺, Cu²⁺, EDTA, citrate and tartrate.
• 4.4. Phosphate ions inhibited it competitively, phenylalanine uncompetitively and urea noncompetitively.
• 5.5. It was heat stable for 60 min at 37°C but labile above 55°C.
• 6.6. Its K_m with p-nitrophenylphosphate was 0.5 mM; its estimated molecular weight was 160,000.
• 7.7. The results are compared with the properties of alkaline phosphatases from the rainbow lizard and man and discussed in terms of a triadic association between the fungus, the lizard and man.

     

Responses of mixed-function oxygenase and antioxidase enzyme system of Mytilus sp. to organic pollution

• 1.1. Mixed-function oxidase (MFO) system components (cytochrome P-450, “418-peak”, cytochrome b₅ and NADPH-cytochrome c (P-450) reductase) and inducible antioxidant enzymes (catalase, Superoxide dismutase (SOD), glutathione peroxidase (GPX) and DT-diaphorase) has been determined in digestive glands of mussels (Mylilus galloprovincialis) collected from three Mediterranean coastal locations, exhibiting an organic pollution gradient.
• 2.2. Cytochrome P-450, the “418-peak”, catalase and SOD showed a good correlation with whole body tissue PAHs and, to a lower extent, with PCBs.
• 3.3. Microsomal NADPH-dependent DT-diaphorase, but not the NADH-dependent microsomal enzyme or the cytosolic DT-diaphorases, was indicated to increase with pollution exposure.
• 4.4. The application of such measurements to environmental monitoring is discussed. Given the magnitude of differences observed, and the state of knowledge on enzyme function and mechanisms of toxicity, a multiparameter approach is considered to offer current and future potential for detecting the impact of organic pollution on bivalve molluscs.

     

Biochemical and physiological aspects of anhydrobiosis

• 1.1. The biochemical and physiological mechanisms which are involved in anhydrobiotic survival have been reviewed.
• 2.2. The physical state of water within hydrated and dehydrated organisms is discussed in relation to the concepts of “free” and “bound” water and to the “vicinal water network model” of Clegg (1979) Cell Associated Water. Academic Press, New York.
• 3.3. Evidence is presented for the replacement of “bound” water by glycerol in dried embryos of Artemia salina, but the role of high glycerol content in the free-living nematode Aphelenchus avenae has yet to be evaluated.
• 4.4. The adaptive significance of trehalose is shown to lie in the fact that because it is a non-reducing sugar, it will not participate in a “browing” reaction between reduced groups of sugars and free amino groups of dry proteins. Trehalose also inhibits “browning” reactions between reducing sugars and dry proteins.
• 5.5. The effect of dehydration on membrane permeability suggests that dried organisms suffer mostly if placed directly into a revival medium due to leakage through structurally deformed membrane systems.
• 6.6. Glycerol and trehalose may interact with lipid membranes and reduce the amount of leakage.
• 7.7. Damage to membranes caused by lipid peroxidation is discussed.
• 8.8. Results suggest that the role of high lipid contents in nematode anhydrobiotes is essentially that of a food reserve, although the morphological distribution of such lipid may be important in maintaining the spatial distribution of body tissues in the absence of bulk water.

     

Potassium contracture of anterior byssus retractor muscle of Mytilus edulis

• 1.1. The potassium contracture in the anterior byssus retractor muscle of Mytilus edulis relaxed spontaneously, and the relaxation was accelerated by 5-HT (10⁻⁶ M), but the contractile activation and inactivation was not affected significantly.
• 2.2. By reducing [Ca]₀, the “activation curve” was shifted downward at higher [K]₀, and the “inactivation curve” was shifted toward lower [K]₀ and the rate of inactivation was increased.
• 3.3. The steady-state inactivation was maintained for at least 2 hr without complete inactivation.
• 4.4. The half-inactivation time was dependent on [K]₀, while the half-relaxation time in the contracture induced by conditioning K solutions was not.

     

Thermal properties for digestive enzymes of a sub-antarctic beetle,hydromedion sparsutum (coleoptera,perimylopidae) compared to those in two thermophilic insects

• 1.1. Proteolytic, lipolytic, amylolytic and cellulolytic activities were studied in adults of the phytophagous beetle, Hydromedion sparsutum, indigenous to the sub-Antarctic island of South Georgia.
• 2.2. Gastric enzyme activities were measured at experimental temperatures of 5–40°C and results were compared with those obtained from two thermophilic insects, Gryllus bimaculatus and Tenebrio molitor.
• 3.3. Protease and lipase activities in Hydromedion were 10–15 times lower than in Gryllus and Tenebrio.
• 4.4. In the temperature range of 5–15°C, α-amylase activity from Hydromedion was only slightly lower than that from Gryllus.
• 5.5. Hydromedion gut homogenates exhibited a distinct cellulolytic activity, even at a low temperature of 5°C.
• 6.6. Cellulolytic activity in the digestive tract of Hydromedion was confirmed by the evolution of ¹⁴CO₂ after consumption of labelled cellulose.
• 7.7. The thermal properties of digestive enzymes agree well with the role of Hydromedion as primary decomposer in its ecosystem.

     

Isolation and characterization of the plasma membrane from slime-forming,encapsulated Streptococcus cremoris

• 1.1. The plasma membrane of slime-forming, encapsulated Streptococcus cremoris from “viili” was isolated in hypotonie conditions in the presence of lysozyme (EC 3.2.1.17) using density gradient centrifugation as the last purification step.
• 2.2. The membrane yield was 15.8% of wet weight cells and the preparation contained 64.4% protein. 19.1% carbohydrate, 5.8% aminosugars, 5.1% RNA and 0.07% DNA.
• 3.3. Buffered 1% (w/v) Triton X-100 solubilized 33.6% of membrane proteins. The number of polypeptides detected by SDS-polyacrylamide gel electrophoresis was 59 when the membrane was isolated without a protease inhibitor and 44 in the presence of a protease inhibitor.
• 4.4. The molecular weights of the polypeptides varied from 13,500 to 100,000.
• 5.5. Ultrathin-layer electrofocusing analysis revealed the range of protein pi values to be between 3.50 and 5.85 concerning 77.3% of proteins and between pI 5.85 and 8.15 concerning 18.2% of proteins.
• 6.6. The isoelectric point of the only basic protein component was 9.3.

     

Metabolic rate and body temperature of adult and juvenile hyrax (Procavia capensis)

• 1.1. Resting metabolic rates (RMR) below thermoneutrality in adult hyrax acclimated to 26, 15 and 10°C remained unchanged, i.e. thermal conductance (K) remained constant.
• 2.2. Conductance in juveniles decreased with acclimation to lower ambient temperatures (T_a).
• 3.3. Body temperature (T_b) dropped by 3.8°C in adults exposed to T_a of 30 – 5°C. The decrease was constant.
• 4.4. Body temperature fell by 1.5°C in juveniles exposed to T_a of 30 – 20°C but stabilized between 20 and 5°C.
• 5.5. The labile T_b, associated with behavioural strategies and lower than predicted RMR, can be seen as an energy-conserving mechanism of particular importance during winter conditions.

     

Two kinds of peripheral afferent neurons in the earthworm reflex arc

• 1.1. Two kinds of neurons were identified in the body-wall longitudinal muscle layer of the earthworm, Amynthas hawayanus, by the simultaneous potential recording and Lucifer Yellow-CH injection method with a single microelectrode.
• 2.2. Both kinds of neurons have their somata, neuntes and longitudinal processes imbedded in the longitudinal muscle layer. Those with two circular processes extending into the third segmental nerve trunk are tentatively named “intra-nerve-trunk” neurons and those with four circular processes extending into four setae shafts are tentatively named “intramural” neurons.
• 3.3. Both kinds of neurons responded to electrical and mechanical stimuli applied in an afferent direction to them.
• 4.4. The “intra-nerve-trunk” neuron decreased its response amplitudes to these stimuli after the third nerve trunk was sectioned in correlation to the response amplitude decrease recorded from the nerve trunk after it was sectioned.
• 5.5. The response amplitude decrease due to denervation implies a nonlinear structure of the earthworm reflex circuits.
• 6.6. The “intramural” neurons are believed to be primary sensory neurons connected to the mechanoreceptors in the setae.

     

Effects of acclimation temperature,season, and time of day on the critical thermal maxima and minima of the crayfish Orconectes rusticus

• 1.1. The critical thermal minima (CTMin) and maxima (CTMax) were determined for field-acclimatized and laboratory-acclimated crayfish (Orconectes rusticus) throughout 1984.
• 2.2. The CTMin and CTMax of field-acclimatized crayfish were seasonally adjusted by 9.7 C and 14.7 C respectively.
• 3.3. Seasonal variation in both tolerance regimes persisted in crayfish acclimated in the laboratory at 5 and 25°C for one week; however, no diel variation existed in either the CTMin or CTMax of laboratory-acclimated crayfish.
• 4.4. Integration of thermal acclimation of the CTMin and CTMax with seasonal conditioning may influence the functional capacities of this species when considered in relation to the seasonal ranges in stream temperature.

     

Heart rate and its cholinergic control in the sole (Solea vulgaris), acclimatized to different temperatures

• 1.1. The effects of thermal acclimatization at 10 and 24°C on heart rate were investigated on unrestrained soles (Solea vulgaris).
• 2.2. The sensitivity of heart rate to temperature changes induced by temperature acclimatization was higher in cold-acclimatized than in warm-acclimatized soles.
• 3.3. Heart rate of cold-acclimatized fish to temperature changes was not affected by blocking the vagal tone with atropine.
• 4.4. After atropine treatment the ability of heart rate to show thermal compensation decreased in warm-acclimatized soles.
• 5.5. It is suggested that the vagus nerve can function differently at different temperatures.

     

Osmoregulation and temperature effects on water loss and oxygen consumption in two species of african scorpion

• 1.1. Aspects of the physiology of two southern African scorpions have been examined. The scorpions are the large desert species Parabuthus villosus (Peters) (Buthidae) and the more mesic, burrowing species Opisthophthalmus capensis (Herbst) (Scorpionidae).
• 2.2. Evaporative water losses were higher in Opisthophthalmus at all temperatures.
• 3.3. Analysis of haemolymph during prolonged desiccation showed good osmotic and ionic regulation in Parabuthus but no regulation in Opisthophthalmus.
• 4.4. Oxygen consumption of Parabuthus was measured after acclimation to 10 and 30°C. Metabolic rates were extremely low but there was no metabolic compensation to increased temperatures.

     

Cyclic AMP stimulation and prostaglandin inhibition of Na transport in freshwater mussels

• 1.1. Pondwater acclimated Carunculina texasensis and Ligumia subrostrata experienced a 230% increase in Na influx when injected with dibutyryl cyclic AMP (0.4mM/l blood).
• 2.2. Theophylline, a phosphodiesterase inhibitor, or indomethacin, an inhibitor of prostaglandin (PG) synthetase, caused a dose dependent stimulation of Na transport.
• 3.3. Prostaglandin E₂ injected into mussels caused an inhibition of Na influx. Arachidonic acid, the precursor of PGE₂, inhibited Na influx or stimulated Na efflux depending on the animal's acclimation conditions.
• 4.4. Chloride transport was unaffected by the drugs used in this study.