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1.
Studies of the enzyme fructose-1,6-bisphosphatase (FBPase) of rainbow trout (Oncorhynchus mykiss) have been undertaken in order to illuminate aspects of skeletal muscle gluconeogenesis in these animals. Maximal activities in crude homogenates of several organs suggest that the liver possesses the greatest FBPase activity on a unit g–1 tissue basis but that the white muscle, owing to its bulk, contributes substantially to whole body FBPase activity. Studies of fructose-6-phosphate-1-kinase (PFK) and FBPase in crude homogenates of several organs suggests an important role for intracellular pH in regulating the relative carbon flux through the FBPase/PFK locus in vivo. Furthermore, a three-step purification scheme is described for trout white muscle FBPase by which a stable and homogeneous (by SDS PAGE) enzyme preparation (isoelectric point = 7.2; molecular weight = 37.6 kd) was obtained. Kinetic studies of the purified enzyme were undertaken at 20°C under conditions reflective of "rest" and "exercise/recovery" intramuscular pH in vivo. Affinity for substrate (F-1,6-P2) was increased (Km = 6.88 versus 2.44 mol 1-–1 as was enzyme activity when pH was lowered from 7.0 to 6.5. Various inhibitor metabolites are identified including F-2,6-P2 (mixed-type inhibitor, Ki = 0.201 mol 1–1, pH 7.0) and AMP (non-competitive inhibitor, Ki = 0.438 mol 1–1, pH 7.0). Inhibition by F-2,6-P2 was strongly alleviated by a reduction in pH from 7.0 to 6.5 (I50 increased from 0.14 to 0.32 mol 1–1). AMP on the other hand was a more potent inhibitor at pH 6.5 but this inhibition was totally reversed under conditions of citrate, NH4 + and AMP typical of muscle during recovery from exercise in vivo. In purified white muscle enzyme preparations, FBPase demonstrated maximal activity at pH 6.5 whereas the optimal pH of PFK was 7.0 or greater. Indeed, it appears from these in vitro data that regulation by metabolite levels as well as pH are required for net FBPase flux in vivo. It is concluded, therefore that trout white muscle FBPase demonstrates the potential to play an important enzymatic role in the control of intramuscular gluconeogenesis in these animals. The results are discussed in relation to present knowledge regarding the metabolic responses of trout white muscle to, and its subsequent recovery from, exhaustive exercise.  相似文献   

2.
Ammonia distribution and excretion in fish   总被引：6，自引：0，他引：6  
This paper reviews the literature concerning ammonia production, storage and excretion in fish. Ammonia is the end product of protein catabolism and is stored in the body of fish in high concentrations relative to basal excretion rates. Ammonia, if allowed to accumulate, is toxic and is converted to less toxic compounds or excreted. Like other weak acids and bases, ammonia is distributed between tissue compartments in relation to transmembrane pH gradients. NH3 is generally equilibrated between compartments but NH4 + is distributed according to pH. Ammonia is eliminated from the blood upon passage through the gills. The mechanisms of branchial ammonia excretion vary between different species of fish and different environments, and primarily involves NH3 passive diffusion and NH4 +/Na+ exchange. Water chemistry near the gill surface may also be important to ammonia excretion, but a more accurate measurement of the NH3 gradient across the gill epithelium is required before a more detailed analysis of NH3 and NH4 + excretion can be made.  相似文献   

3.
Gas transfer and blood acid-base status in the blood-perfused trout head preparation (in vitro) were compared with intact trout (in vivo) fitted with oral membranes, dorsal aortic, ventral aortic, and opercular cannulae. Gas transfer rates were calculated from both arterial-venous blood differences and inspired-expired water differences using the Fick method. Oxygen uptake and carbon dioxide excretion were lower, while ammonia excretion was higher, in the blood-perfused head relative toin vivo rates. Gas transfer rates calculated from water were consistently greater than rates calculated from blood, the difference being greaterin vitro compared toin vivo. We conclude that the inadequacy of O2 and CO2 transfer in the blood-perfused head was not due to abnormal gill diffusive conductance, but was more likely related to the reduced magnitude of the blood-to-water O2 and CO2 diffusion gradients, low hematocrit, and decreased perfusion flow rate . Under the conditions of the present study, the blood-perfused trout head is not a suitable preparation for the study of oxygen transfer. We conclude this preparation is useful for evaluating branchial carbon dioxide or ammonia transfer only when comparable measurements or manipulations cannot be made on intact fish.  相似文献   

4.
NAD+-linked isocitrate dehydrogenase was found in the brain, heart, gills, kidney, liver and muscle of trout, and in the liver and muscle of eel. A complex homogenization buffer containing 1 mM ADP, 5 mM MgSO4, 5 mM citrate and 40% glycerol is required for retrieval of significant amounts of stable enzyme. The highest activities were found in brain of trout and the lowest in white muscle of trout and eel. The enzyme was partially purified from frozen trout heart to a final activity of 0.04 M/min/mg protein, and the kinetic properties of this partially purified enzyme were studied. The enzyme requires either Mn2+ or Mg2+ for activity, higher activities being observed with Mn2+. Saturation kinetics for DL-isocitrate were sigmoidal, apparent S0·5=8.2±0.6 mM and nH=1.8±0.2, in the absence of ADP, changing to hyperbolic, apparent S0·5=1.4±0.3 mM and nH=1.0, with 1 mM ADP added. Citrate and Ca2+ were found to activate the enzyme to a small extent. NADH strongly inhibited the enzyme, I50=3.7±0.5 M. ATP was also found to be an inhibitor, I50=7.2±1.4 mM. These properties are consistent with the role of the enzyme as a major control site of the tricarboxylic acid cycle.  相似文献   

5.
Soft water acclimated (Ca2+ 0.02 mM; Na+ 0.03 mM; K+ 0.01 mM; pH 7.0), cannulated brown trout (Salmo trutta) were exposed to various pH and aluminium (Al) regimes (pH 7.0, pH 5.0, pH 5.0 plus Al: 50, 25, and 12.5 g l–1) for up to 5 days in order to determine (i) the sublethal concentration of Al at pH 5.0 for this species (ii) their ionoregulatory and respiratory status. No mortality or physiological disturbances were evident at pH 7.0 or pH 5.0. All trout died within 48 h at pH 5.0 in the presence of Al at 50 g l–1 and 67% died over the 5 day period at pH 5.0 in the presence of Al at 25 g l–1. Fish at these lethal Al concentrations showed significant decreases in arterial blood oxygen content (CaO2) but no changes in plasma osmolarity or the concentrations of plasma Na+, K+ and Cl–. Physiological disturbance was more marked at the 50 g l–1 Al concentration. The surviving fish at 25 g l–1 showed few signs of physiological recovery while continually exposed to this regime. No fish died during the exposure to water of pH 5.0 containing 12.5 g l–1 Al, but physiological disturbance was still apparent. These sublethally-stressed trout showed a transient decline in the plasma concentrations of Na+ and Cl–1. Although CaO2 decreased, recovery was evident. The data suggest that in the brown trout, environmental Al concentration is as important as pH and calcium concentration in determining the physiological status of the fish.  相似文献   

6.
The effect of sulfide on K+ influx pathways was measured in red blood cells (RBCs) of sulfide-sensitive rainbow trout (Oncorhynchus mykiss) and sulfide-tolerant crucian carp (Carassius carassius). In trout RBCs, maximal inhibition of Na+, K+-ATPase was attained at 10 mol l–1 sulfide and amounted to 32% without being influenced by pH between 6.7 and 8.3. Ouabain-resistant K+ influx in the absence and presence of sulfide was insignificant at pH values between 6.7 and 7.7. At higher pH values ouabain-resistant K+ influx increased, but was inhibited to about 15% by 30 mol l–1 sulfide. In RBCs of crucian carp neither Na+, K+-ATPase nor ouabain-resistant K+ influx were affected by sulfide concentrations up to 850 mol l–1. Differences in sulfide-sensitivity of K+ influx between both species can be based upon different properties of the membrane transporter themselves. The reduced Na+, K+-ATPase activity in trout RBCs may also result from a slightly reduced (by 9%) ATP level after sulfide exposure. In addition, intracellular sulfide concentrations were higher in trout RBCs as compared to crucian carp. In trout, intracellular sulfide concentrations reached extracellular levels within 5 min of incubation whereas sulfide concentrations in crucian carp RBCs remained about 2-fold lower than extracellular concentrations. Although the physiological basis of sulfide-insensitive K+ influx in crucian carp RBCs is currently unknown it may contribute to the extremely high sulfide-tolerance of this species.  相似文献   

7.
Four successive life stages (zoea-III, zoea-IV, zoea-V and megalopa) of the Chinese mitten-handed crab, Eriocheir sinensis (H. Milne-Edwards), were exposed to ammonia in a series of short-term bioassays with the static-renewal method at 22°C, pH 8.0 and 25%o salinity. The greatest sensitivity was observed in the zoea-III stage. The 24-h LC50 values for zoea-III, zoea-IV, zoea-V and megalopa were 32.8, 73.1, 84.0 and 90.1 mg L?1 for NH3+ NH4+, and 1.11, 2.36, 2.77 and 3.18 mg L?1 for NH3, respectively. The 72-h LC50 values for zoea-III, zoea-IV and zoea-V were 11.9, 23.6 and 38.2 mg L?1 for NH3+ NH4+, and 0.40, 0.76 and 1.26 mg L?1 for NH3, respectively. The 96-h LC50 values for megalopa were 37.3 mg L?1 for NH3+ NH4+ and 1.31 mg L?1 for NH3. It was found that ammonia tolerance increased with larval development from zoea-III to megalopa, especially from zoea-III to zoea-IV and from zoea-IV to zoea-V. A comparison of safe levels of ammonia among the different life stages indicated that all stages were significantly different with respect to safe levels of ammonia (P < 0.05) except zoea-V and megalopa, which had the highest safe levels. In general, both the larvae and juveniles of E. sinensis are less resistant to ammonia than those of other crustacean species studied so far.  相似文献   

8.
It is yet unclear whether sub‐lethal ammonia‐N levels cause irreparable damage to aquatic crustaceans, or if recovery is possible, the potential factors involved. The aim was to investigate the effect of 0.706 and 2.798 mmol L?1 ammonia‐N exposure on the haemolymph osmolality, Na+, K+, Ca2+, pH, ammonia‐N, total haemocyte counts (THC) and gill histopathology of Portunus pelagicus juveniles at 0, 3, 6, 12, 24 and 48 h respectively. Following 48 h, crabs were transferred to pristine seawater allowing a recovery period up to 96 h and similarly measured. In addition moribund crabs, induced from lethal ammonia‐N levels of 7.036 and 10.518 mmol L?1, were measured for haemolymph osmolality/ions and pH levels. The results demonstrate that despite severe gill damage within 6‐ and 1 h of 0.706 and 2.798 mmol L?1 ammonia‐N exposure, respectively, no significant change (P>0.05) in the haemolymph osmolality, Na+, K+, Ca2+ or pH levels occurred or by ammonia‐N‐induced morbidity. Although the gills can completely recover within 24 and 48 h post exposure to 0.706 and 2.798 mmol L?1 ammonia‐N, respectively, likely facilitated by significant haemocyte increases (P<0.05) within the haemolymph and gill lamellae, dependent factors were the previous ammonia‐N concentration and recovery duration while individual variability was also noticed.  相似文献   

9.
The purification of trypsin from the common kilka (Clupeonella cultriventris caspia) viscera (pyloric caeca) resulted in a 28.3-fold increase and 12% yield by ammonium sulfate precipitation (30–60%), Sephadex G-75, and DEAE-cellulose chromatography. Trypsin showed a molecular weight of 23.2 kDa and appeared as a single band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), native-PAGE, and zymography. The trypsin had optimal activity at pH 8.0 and 60°C for the hydrolysis of α-N-benzoyl-DL-arginine-ρ-nitroanilide hydrochloride (BAPNA) substrate. Trypsin was stable up to 50°C and at pH range of 7.0–10.0. Activity was significantly inhibited by soybean trypsin inhibitor (SBTI) and N-ρ-tosyl-L-lysine-chloromethylketone (TLCK) inhibitors (p < 0.05). The enzyme was relatively stable toward oxidizing agents, retaining 59.7 and 98.0% of its initial activity after 1 h incubation in the presence of 15% H2O2 and 1% sodium perborate, respectively. Trypsin was significantly activated by surfactants and Ca2+, Mg2+, and Mn2+ and inactivated by Fe2+, Zn2+, Cu2+, Al3+, Ba2+, and Co2+ (p < 0.05). Nevertheless, Na+ and K+ had no significant effect on trypsin activity (p > 0.05). The purified trypsin showed significantly higher catalytic efficiency (kcat/Km) than porcine pancreatic trypsin against BAPNA and N-α-p-Tosyl-L-arginine methyl ester hydrochloride (TAME) substrates (p < 0.05).  相似文献   

10.
The effects of transporting silver catfish (Rhamdia quelen) for 6 h in plastic bags containing 0 (control), 30 or 40 µL/L of essential oil (EO) from Lippia alba leaves were investigated. Prior to transport, the fish in the two experimental groups were sedated with 200 µL/L of EO for 3 min. After transport, dissolved oxygen, carbon dioxide, alkalinity, water hardness, pH, temperature and un-ionized ammonia levels in the transport water did not differ significantly among the groups. However, total ammonia nitrogen levels and net Na+, Cl? and K+ effluxes were significantly lower in the groups transported with EO of L. alba than those in the control group. PvO2, PvCO2 and HCO3 ? were higher after transporting fish in 40 µL/L of EO of L. alba, but there were no significant differences between groups regarding blood pH or hematocrit. Cortisol levels were significantly higher in fish transported in 30 µL/L of EO of L. alba compared to those of the control group. The metabolic parameters (glycogen, lactate, total amino acid, total ammonia and total protein) showed different responses after adding EO to the transport water. In conclusion, while the EO of L. alba is recommended for fish transport in the conditions tested in the present study because it was effective in reducing waterborne total ammonia levels and net ion loss, the higher hepatic oxidative stress in this species with the same EO concentrations reported by a previous study led us to conclude that the 10–20 µL/L concentration range of EO and lack of pre-sedation before transport are more effective.  相似文献   

11.
Reuse of fish effluent for the culture of marine shrimp, such as Pacific white shrimp, Litopenaeus vannamei, could provide an opportunity for the US shrimp farming industry to ease constraints (e.g., environmental concerns and high production costs) that have limited them in the past. In this study under laboratory‐scale conditions, the feasibility of culturing L. vannamei in effluents derived from a commercial facility raising tilapia in recirculating aquaculture systems (RAS), supplemented with various salt combinations, was compared to the shrimp’s survival and growth in well water supplemented with 17.6 (control) and 0.6 (freshwater treatment) g/L synthetic sea salt. Three independent trials were conducted in RAS in which survival and growth in the control, the freshwater treatment, and two effluent treatments were compared. Water quality during this study was within safe levels and no differences (P < 0.05) between treatments were observed for dissolved oxygen, nitrite, pH, total ammonia nitrogen, and temperature. However, average nitrate and orthophosphate levels were consistently more than an order of magnitude greater in the effluent treatments compared to the control and the freshwater treatments. Survival and growth of shrimp over 6‐wk periods did not vary significantly between the control and the freshwater treatments; however, shrimp tested in the tilapia effluents often exhibited significant effects (P < 0.05) depending on the salts added. In the low‐salinity waters, correlations (P < 0.05) were observed between Ca2+, Mg2+, Ca2+ and Mg2+, K+, Na+ : K+ and Ca2+ : K+, and shrimp survival and growth. The results of this study revealed that L. vannamei can be raised in tilapia effluent when supplemented with synthetic sea salt (0.6 g/L), CaO (50 mg/L Ca2+), and MgSO4 (30 mg/L Mg2+).  相似文献   

12.
A novel marine origin Bacillus subtilis strain H1 isolated from a shrimp culture pond effectively removed NH4+‐N, ‐N and ‐N, with a maximum ammonium, nitrite and nitrate removal rate of 2.35 mg NH4+‐N L?1 hr?1 per OD, 9.64 mg ‐N L?1 hr?1 per OD and 0.75 mg ‐N L?1 hr?1 respectively. The gas chromatography–isotope ratio mass spectrometry results indicated that N2O was emitted when 15NH4Cl, Na15NO2 or Na15NO3 was used. Additionally, N2 was also produced when Na15NO2 was used. Single‐factor experiments suggested that the optimal conditions for NH4+‐N and ‐N removal were glucose as a carbon source, C/N 15, initial pH 7.5, 30 g/L NaCl, 28°C and a shaking speed of 160 rpm. Orthogonal tests showed that the optimal conditions for NH4+‐N removal were C/N 15, pH 9, 10 g/L NaCl and shaking speed 160 rpm when ammonium chloride was used as the substrate. The optimal conditions for ‐N removal were C/N 10, pH 6, 10 g/L NaCl and a shaking speed of 160 rpm when sodium nitrite was used as the substrate. In summary, B. subtilis strain H1 had highly efficient aerobic nitrifying–denitrifying ability and high adaptability, suggesting that it is potentially valuable to marine aquaculture.  相似文献   

13.
Starry flounder (Platichthys stellatus) were cannulated and a bolus of 9 Ci14C-creatine in saline was injected into the caudal vein. The fish were sacrificed at intervals ranging from 1h to 36d after label injection. Creatine pool size (PCr+Cr) and creatinine (Crn) content in blood, muscle, gills and liver were analyzed and specific activities (SA) determined.Mean concentrations of PCr+Cr/Crn in PCA-extracts of muscle, gills, liver and blood of experimental fish (at rest) were 38.1/2.40, 4.1/0.25, 5.6/0.45 and 0.3/n.d. mol.g–1 respectively.Within 10 min, plasma SA had decreased by approximately 90%. In white muscle, the rate of14C–Cr appearance as well as label disappearance was slow compared to gills and liver. In fish examined 36d postinjection, mean SA in muscle had decreased to 23% of maximum SA which occurred 24h after injection.14C–Cr was incorporated into the liver tissue at a very high rate, SA being two orders of magnitude higher in liver than in white muscle. Over the first 6d, retention of label was observed in liver; after 36d only 3% of the original label was detected.Creatine pool size (PCr+Cr) in white muscle decreased with food deprivation. In flounder sacrificed after 36d, PCr+Cr was only 52% that of fed control fish, suggesting that creatine or precursors for its biosynthesis are supplied with the diet.  相似文献   

14.
Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD+ oxidoreductase (decarboxylating)] was purified from herring skeletal muscle to a specific activity of 8.2 mol/min/mg. The purification procedure involved chromatography on DEAE-cellulose, Red Agarose and a Sephacryl S-300 with a final recovery of 38% of enzyme activity. This enzyme catalyzes the oxidative decarboxylation of malate in the presence of either NAD or NADP in the presence of Mn2+. Some kinetic characteristics of this enzyme were determined. The pH optimum of activity is 7.0. ATP was shown to be a competitive inhibitor with malate. The inhibition by ATP displayed hyperbolic competitive kinetics with a Ki (ATP) of 0.28 mM in the presence of NAD and 0.75 mM in the presence of NADP. Fumarate reversed ATP inhibition.In vivo, regulation of NAD(P)-dependent malic enzyme might respond to changing levels of mitochondrial ATP and fumarate with the enzyme undergoing kinetic activation by an increase in the concentration of mitochondrial fumarate which could reverse enzyme inhibition by ATP.  相似文献   

15.
Gastric acid secretion from isolated cod stomach mucosa was measured using a pH-static titration method. A basal acid secretion rate (BASR) of 6.0±0.6 nEqH+min–1cm–1 was measured when using 0.9% NaCl as luminal solution. There was a dose-dependent increase in response to histamine between 0.12 and 0.20 M (EC50=0.15 M), above which gastric acid secretion plateaued at 13.5±1.8 nEqH+min–1cm–1. Ranitidine, a H2-receptor antagonist, completely blocked the stimulatory effect of histamine and reduced the BASR. The H1-receptor antagonist, clemastine, did not inhibit the response to histamine. Acid secretion rates decreased significantly when the pH of the luminal side of the mucosa was lowered from pH 5.75 to pH 4.50, indicating that a negative feedback mechanism was operating. Histological staining showed that oxynticopeptic cells were uniformly distributed throughout the cardiac stomach.It is concluded that the acid secretion in the isolated stomach mucosa of cod can be measuredin vitro with a pH-static titration method. The method was used to demonstrate that the BASR is downregulated by a decrease in pH. Furthermore, we conclude that the histamine receptor in the cod stomach mucosa resembles the mammalian H2-receptor and that histamine is secreted under basal conditions.  相似文献   

16.
The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg–1 fish·h–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g–1·ml–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min–1·g–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min–1·g–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.  相似文献   

17.
The ability of carp to recover from nitrite-induced methaemoglobinaemia and disturbances in potassium balance and cell volume was studiedin vivo andin vitro. Nitrite accumulated to a plasma concentration of 3 mM during 2 days of nitrite exposure was eliminated from the plasma within 2–3 days in clean water. The nitrite-induced methaemoglobinaemia disappeared after 3 days of recovery. During nitrite exposure, K+ was lost from the red blood cells (RBCs) and from skeletal muscle tissue, which led to reduced cell volume and an extracellular hyperkalaemia. Extracellular [K+] rose less than predicted if lost K+ had remained in the extracellular space, suggesting further transport of K+ to the environment. The intracellular K+ and water content were restored after few days of recovery in clean water, but this was paralleled by development of an extracellular hypokalaemia. This shows that intracellular K+ balance was reestablished at the expense of the extracellular compartment, and supports that an overall K+ deficit resulted from K+ loss to the environment during nitrite exposure. Ventricle tissue differed from skeletal muscle and RBCs by not loosing K+ and by having increased sodium and water contents during nitrite exposure. These changes were corrected by recovery in nitrite-free water. In vitro addition of nitrite to blood with low O2 saturation induced metHb formation and RBC K+ efflux. Subsequent reduction of metHb to functional Hb was similar in blood with low and high O2 tension. A net re-uptake of K+ was observed only in RBCs with low O2 saturation and when metHb reached low values.  相似文献   

18.
Static-renewal bioassays were performed to evaluate the acute toxicity of ammonia to Eriocheir sinensis (H. Milne-Edwards) at three growing stages, namely zoea-I, zoea-II, and juvenile (0.06 g wet weight per crab). The 24 h LC50 values were 13.3, 20.2, and 109.3 mg (NH3+ NH4+) 1?1 (0.47, 0.71, and 3.10 mg NH3 I?1), the 48 h LC50 values being 6.8, 10.3, and 60.9 mg (NH3+ NH4+) 1?1 (0.24, 0.36, and 1.73 mg NH31?1), while the 72 h LC50 values were 5.7, 7.6, and 45.3 mg (NH3+ NH4+) 1?1 (0.20, 0.27, and 1.29 mg NH3 1?1) for zoea-I, zoea-II, and juveniles, respectively. The 96 h LC50 value for juveniles was 31.6 mg (NH3+ NH4+) 1?1(0.90 mg NH31?1). It was evident that the tolerance to ammonia increased during the same exposure time as the larvae developed to juveniles and decreased with prolonged exposure time. Compared with larvae, juveniles were more sensitive to the fluctuation of ambient ammonia concentrations in the certain range within which partial kills took place. The ‘safe level’ of ammonia based on the 96 h LC50 value and an application factor of 0.1 was 3.16 mg (NH3+NH4+)1?1 (0.09 mg NH3 1?1) for juveniles and those for zoea-I and zoea-II were 0.57 and 0.76 mg (NH3+ NH4+) 1?1 (0.02 and 0.03 mg NH3 1?1) based on 72 h LC50 values.  相似文献   

19.
The effects of severe experimental anaemia on red blood cell HCO3 – dehydrationin vitro were examined in rainbow trout,Oncorhynchus mykiss. After 5 days of anaemia (haematocrit=4.9±1.1%) induced by intraperitoneal injection of phenylhydrazine hydrochloride, fish displayed elevated arterial CO2 tensions (anaemic PaCO2=3.19±0.42 torrvs. control PaCO2=1.35±0.17 torr) and a significant acidosis (anaemic pHa=7.73±0.04vs. control pHa=7.99±0.04). However, after 15–20 days of anaemia (hct=6.6±0.8%) induced by blood withdrawal, the arterial CO2 tension was significantly lower than the control value, suggesting that physiological adjustments occurred within this time period to compensate for the lowered haematocrit. Compensation probably did not involve alterations in ventilation, which was unaffected by 5 days of anaemia (anaemic ;w=786±187 ml min–1 kg–1 vs. control ;w=945±175 min–1 kg–1), based on indirect Fick principle measurements.Potential adaptations to longer term anaemia at the level of the red blood cells were investigated using a radioisotopic HCO3 – dehydration assay. Owing to the difference in haematocrits, the HCO3 – dehydration rate for blood from anaemic fish was significantly lower than that for control fish following equilibration at the same CO2 tension. This difference was eliminated when HCO3 – dehydration rates were measured on blood samples adjusted to the same haematocrit, a result which implies that the intrinsic rate of CO2 excretion at the level of the red blood cell was not up-regulated during anaemia. The difference was also eliminated by equilibrating the blood samples with CO2 tensions appropriate for the group from which the sample was obtained,i.e., PCO2=1.4 torr for control samples and PCO2=3.2 torr for anaemic samples; each at the appropriate haematocrit. It is concluded that the elevated PaCO2 helps to reset CO2 excretion to the control level, but that some additional physiological adjustment occurs to lower the PaCO2 after 15–20 days of anaemia.  相似文献   

20.
The presumptive Na+/H+ exchange sites of trout and eel erythrocytes were quantified using amiloride-displaceable 5-(N-methyl-N-[3H]isobutyl)-amiloride (3H-MIA) equilibrium binding to further evaluate the mechanisms of i) hypoxia-mediated modifications in the trout erythrocyte -adrenergic signal transduction system and ii) the marked differences in the catecholamine responsiveness of this system between the trout and eel. MIA was a more potent inhibitor of both trout apparent erythrocyte proton extrusion (IC50 = 20.1 ± 1.1 mol l–1, N = 6) activity (as evaluated by measuring plasma pH changes after addition of catecholamine in vitro) and specific 3H-MIA binding (IC50 = 257 ± 8.2 nmol l–1, N = 3) than amiloride, which possessed a proton extrusion IC50 of 26.1 ± 1.6 mol l–1 (N = 6) and a binding IC50 of 891 ± 113 nmol l–1 (N = 3). The specific Na+ channel blocker phenamil was without effect on adrenergic proton extrusion activity or specific 3H-MIA binding. Trout erythrocytes suspended in Na+-free saline and maintained under normoxic conditions possessed 37,675 ± 6,678 (N = 6) amiloride-displaceable 3H-MIA binding sites per cell (Bmax, presumptive Na+/H+ antiporters) with an apparent dissociation constant (KD) of 244 ± 29 nmol l–1 (N = 6). Acute hypoxia (PO2 = 1.2 kPa; 30 min) did not affect the KD, yet resulted in a 65% increase in the number of presumptive Na+/H+ antiporters. Normoxic eel erythrocytes, similarly suspended in Na+-free saline, possessed only 17,133 ± 3,716 presumptive Na+/H+ antiporters (N = 6), 45% of that of trout erythrocytes, with a similar KD (246 ± 41 nmol l–1, N = 6). These findings suggest that inter- and intra-specific differences in the responsiveness of the teleost erythrocyte -adrenergic signal transduction system can be explained, in part, by differences in the numbers of Na+/H+ exchange sites.  相似文献   

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