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1.
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   

2.
The substrate specificity of brain acetylcholinesterase (AChE) in adult Carassius auratus fish and its sensitivity to carbamate insecticides were investigated in vitro. The results showed that the order of four substrates hydrolyzed by brain AChE in C. auratus was acetylthiocholine iodide > β-methylthiocholine iodide > propionylthiocholine iodide > butyrylthiocholine iodide, and the maximum velocity (V max) of AChE hydrolyzing acetylthiocholine iodide (ATCh) was the highest among the four substrates, and the V max values were 0.067 and 0.082 mmol min−1 mg−1 for male and female fish respectively. But their Michaelis–Menten constants (K m) were the lowest, only 0.071 and 0.072 mmol/l respectively. Compared with other carbamate insecticides, the sensitivity of brain AChE to carbofuran was the highest and the IC50 values were 1.04 × 10−6 mol/l for females and 1.17 × 10−6 mol/l for males. The inhibitory tendencies of eserine, methomyl, and aldicarb to brain AChE were very similar, and the percentage inhibition increased with time at the concentration of 1 × 10−6 mol/l. The order of inhibition potential of the three inhibitors from the highest to the lowest was eserine, aldicarb, and methomyl.  相似文献   

3.
Plasma cortisol levels and the number (Nmax) and affinity (Kd) of specific hepatic cortisol-binding sites were determined in rainbow trout subjected to chronic confinement stress for 14 days. Confinement significantly elevated plasma cortisol levels to 47.3 ± 13.5 ng ml–1 within 24h and although levels declined to 8.0 ± 3.0 ng ml–1 after 14 days, they were significantly higher throughout than levels in unstressed control fish (< 2.0 ng ml–1). There was a 60% reduction in cytosolic Nmax in stressed fish during the first 24h of confinement (35.8 ± 7.9 cf. 129.0 ± 15.2 fmol mg–1 protein), a decline which was sustained at 7 days after the onset of stress but, although numbers of binding sites in the liver of stressed fish were still lower than in unstressed fish, the difference was no longer significant after 14 days of confinement. There was an accompanying significant rise in the Kd of cortisol binding in stressed fish during confinement, from 4.0 ± 0.6 nM at time 0 to 8.4 ± 0.8 nM after 24 h confinement. This increment in Kd was sustained at a level significantly higher than in control fish throughout the 14 day confinement period, despite marked reductions in cortisol levels and increases in Nmax in stressed fish. Throughout the study, specific binding of cortisol could not be consistently detected in high-salt nuclear extracts from stressed or unstressed fish, suggesting either that high-affinity binding sites for cortisol were absent from these preparations, that receptors were present but unable to interact with ligand because they were occupied, or that receptors were present but not being extracted. These possibilities were investigated using a range of extraction procedures, by varying the temperature of incubation, by employing dexamethasone as ligand and by examining binding in purified, intact, nuclei. Estradiol was employed as a methodological control throughout and substantial amounts of specific estradiol binding were detected in all compartments and preparations. Specific cortisol-binding sites were detected in intact nuclei of both stressed and unstressed fish, at levels an order of magnitude lower than estradiol binding in the same preparations. These data demonstrate that activation of the pituitary-interrenal axis leads to significant changes in the nature of target-tissue binding of cortisol in rainbow trout, and reveal a clear difference in the subcellular distribution of binding-sites for estradiol and cortisol, which reflects the situation in mammalian tissues.  相似文献   

4.
The activity and molecular forms of acetylcholinesterase (AChE, EC 3.1.1.7) were characterized in the brain, heart, white skeletal muscle and liver of the Siberian sturgeon (Acipenser baeri), European eel (Anguilla anguilla), wels (Silurus glanis), common carp (Cyprinus carpio) and silver carp (Hypophthalmichthys molitrix), belonging in four families. The brain had the highest (between 183.2±10.6 and 1361.2±189.7 mU mg-1 protein and between 4.7±0.4 and 23.5±2.7 U g-1 wet tissue) and the liver the lowest (between 25.3±1.9 and 126.5±22.7 mU mg-1 protein and between 1.0±0.1 and 4.5±0.5 U g-1 wet tissue) activity in all these fish, with the exception of the wels, where the heart contained the smallest amount of AChE (26.2±7.5 mU mg-1 protein and 1.2±0.3 U g-1 wet tissue). The highest tissue AChE activity was found in the carp and lowest in the sturgeon. The solubility properties and molecular forms of AChE in the four tissues were studied by extraction in high-salt medium (1.0 M NaCl) with and without the detergent Triton X-100 (0.5%, v/v). The proportions of detergent-soluble (DS) and salt-soluble (SS) forms of AChE varied considerably from one species to another, but a general tendency could be observed: the proportion of DS AChE was generally higher in the brain (between 70.4±4.1 and 82.5±1.2%) and lower in the heart (between 12.0±2.0 and 51.3±2.3%) and skeletal muscle (between 16.0±2.0 and 64.4±0.7%). Velocity sedimentation centrifugation revealed that most tissues contained the G4 or A4 form. Three tissues (brain, heart, and liver) of the Siberian sturgeon contained a relatively high proportion of the G1 form (between 24.3±2.4 and 28.6±4.9%), while the skeletal muscle of the wels contained only the most complex type of AChE, i.e., A12. Amongst the tissues studied, the brain and liver mostly contained amphiphilic globular forms of the G4 type, while the heart and skeletal muscle were rich in asymmetric forms (A4 and A12). It was concluded that the differences revealed in the activities of AChE and in the distribution of its molecular forms may be connected with the variation in behavioral habits (feeding and swimming patterns), and the developmental stage of the nervous system.  相似文献   

5.
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   

6.
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   

7.
In this investigation, the effect of dietary administration of curcumin on the healing of skin wound in fish, Labeo rohita, has been reported. Fish were divided into three groups: control group (fish without skin wound), sham group (fish with skin wound without curcumin treatment) and curcumin‐treated group (fish with skin wound and subjected to dietary administration of 1% curcumin). Experiments were conducted for 30 days to assess the healing of skin wounds at different time intervals using scanning electron microscopy, histology, and mucopolysaccharide and enzyme histochemistry. In the curcumin‐treated group, healing of skin wounds was found to be enhanced than in the sham group as indicated by early restoration of morphology of the surface layer of epithelial cells; the density of the mucous goblet cells; the density of club cells in epidermal layer; and early granular tissue formation, collagen deposition and tissue remodelling in dermal layer. Furthermore, peroxidase and catalase enzyme activity showed increased endogenous defence system in the curcumin‐treated group compared with the sham group. It could be concluded that dietary administration of curcumin is beneficial in rapid healing of skin wounds in fish. Early healing of wounds could be considered to prevent the invasion of pathogens and to maintain the integrity of the surrounding tissue.  相似文献   

8.
Studies were conducted to determine the absorption, excretion and requirement of dietary phosphorus (P) by Atlantic salmon (Salmo salar). Triplicate groups of salmon parr, initial weight 15 ± 0.5±g , were fed, diets containing 4, 5, 6, 7, 8, 9, 11 and 13 mg P and 20 KJ of digestible energy (DE) per±g of diet (dry matter basis, DMB) to satiation for 16 weeks. The basal diet containing 4 mg P g-1 (0.15 mg available P per KJ DE) was supplemented with graded levels of calcium phosphate, Ca(H2PO4)2H2O to formulate the eight experimental diets. The fish were reared in fresh water at a temperature of 15 °C on a 12 h photoperiod. Vertebrae ash increased from 316 to 516 mg g-1 fat-free dry matter with an increase in dietary P content. P requirement was estimated by using a four-parameter sigmoidal equation. The data suggests that a diet of 0.28 mg available P per KJ DE is needed for Atlantic salmon. Phosphate and calcium levels in plasma and bone increased, whereas levels of magnesium and liver cholecalciferol decreased, with an increase in dietary P.Phosphate excretion in urine and apparent availability of P were determined in deficient and replete fish. In deficient fish, the urine phosphate concentration was 0.10 mmol L-1 before feeding and 0.25 mmol l-1 after feeding, whereas in replete fish these concentrations were 1.09 and 5.11 mmol l-1, respectively. The increase in urine phosphate concentration was higher in replete fish than in deficient fish, however, the apparent absorption of P was found to be significantly lower in replete fish than in deficient fish. These results suggest that similarly to terrestrial vertebrates, P homeostasis in Atlantic salmon is controlled by absorption in the intestine, conservation in the kidney and storage in the bones.  相似文献   

9.
The activities of Phase I and Phase II biotransformation enzymes in the livers of yearling coho salmon (Oncorhynchus kisutch), were measured biweekly from February until the release date from the hatchery in mid-June, in order to observe any alterations in baseline levels during smoltification. Peak enzyme activities occurred in February and March and then declined through to June. Total cytochrome P450 levels ranged from 0.024±0.009 to 0.095±0.010 nmol mg-1 microsomal protein, ethoxyresorufin O-deethylase activity ranged from 2.74±0.75 to 9.94±0.85 pmol min-1 mg-1 microsomal protein, and glutathione S-transferase activity ranged from 0.07±0.01 to 0.33±0.01 μmol min-1 mg-1 cytosolic protein during this period. Following an intraperitoneal injection of [3H]benzo[a]pyrene (B[a]P), elimination occurred rapidly (>71% excreted into the bile within 24h) from February to June. Although the distribution of B[a]P in tissues changed through the sampling period, the highest leels of B[a]P-derived radioactivity were found in the liver, bile and fat. Analysis of the bile revealed that 55 to 63% of the radioactivity was Phase I metabolites, 16 to 24% glucuronide conjugates, 8% sulfate conjugates, 7% other conjugates and 6% aqueous-soluble metabolites. These findings suggest that the transformation from freshwater adapted coho ‘parr’ to ‘smolts’, can significantly alter biotransformation enzyme activities and the distribution and elimination of xenobiotics such as benzo[a]pyrene in these fish. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

10.
The pattern of changes of activity of the urea cycle enzymes and the rate of urea-N excretion were studied in the perfused liver of an Indian air-breathing ureogenic walking catfish, Clarias batrachus. The liver was perfused with different concentrations of NH4Cl for a period of 60 min to determine the role of ammonia for stimulation of hepatic ureogenesis and the threshold level of ammonia loading needed to cause such stimulation. Both the urea-N excretion and the ammonia uptake by the perfused liver were found to be a saturable process. Ammonia accumulated significantly in the liver infused with 1.25 moles g liver –1 min–1 of NH4Cl, followed by a maximum accumulation of about 28.5 moles g wet wt–1 with the infusion of 5.08 moles g liver–1 min–1. The Vmax of the urea-N excretion (0.47 mol g liver–1 min–1) was obtained with the addition of 5.08 moles g liver–1 min–1 of NH4Cl. Both the tissue and the specific activity of the urea cycle enzymes, except ornithine transcarbamylase and arginase, were stimulated significantly with the infusion of either 1.25 or 5.08 moles g liver–1 min–1 of NH4Cl. Maximum stimulation of tissue activity of carbamoyl phosphate synthetase (about 120%) was seen with the infusion of 5.08 mol g liver–1 min–1, and for argininosuccinate synthetase (about 135%), and argininosuccinate lyase (about 50%) with the infusion of 10.81 mol g liver–1 min–1 of NH4Cl. Higher accumulation of ammonia of about 10–15 mol g wet wt–1 from the physiological level in the perfused liver while infusing with NH4Cl was suggested to be one of the major causes of stimulation of ureogenesis. The presence of such physiological adaptive strategy is probably necessary in this unique group of air-breathing walking catfish to survive under hyper-ammonia stress in their normal habitat or while living outside water or while burrowing inside mud.  相似文献   

11.
The present study investigated the relationships between egg viability and ovarian fluid composition, egg physiology and egg metabolism in lake trout, Salmo trutta lacustris, to obtain biomarkers for egg quality determination. The ovarian fluid pH, protein levels and activities of aspartate aminotransferase and -d-glucuronidase were significantly correlated with egg viability expressed as the number of eyed stage embryos. Regression models demonstrated that an ovarian fluid pH between 8.44 and 8.57, protein levels below 235.56 mg 100 ml–1ovarian fluid, aspartate aminotransferase activity below 31.65 m min–1 l–1ovarian fluid and -d-glucuronidase activity below 8.62 m min–1 l–1 ovarian fluid characterized egg batches with high viability (80%).The increase in the egg wet weight during water hardening was also significantly correlated with the number of eyed stage embryos, and egg batches with high egg viability (80%) increased in wet weight by 13% during water hardening.From the investigated metabolic parameters the number of eyed stage embryos was significantly correlated with activities of NADP-dependent isocitrate dehydrogenase (egg viability 80% at 2.07 nM min–1 mg–1 protein) and NAD-dependent malate dehydrogenase (egg viability 80% at 47.25 nM min–1 mg–1 protein), with the respiration rate (egg viability 80% at 8.71 nM min–1 mg–1 protein), with the ratio of NADH to NAD levels (egg viability 80% 0.872), with the levels of free, non-esterified fatty acids (egg viability 80% 72.34 g mg–1 protein), and the ratio of non esterified to esterified fatty acids (egg viability 80% at 0.749). Also, subjective and visual control methods were described to distinguish between batches with viable and non viable eggs.  相似文献   

12.
Three commercially available fatty acid enrichment emulsions (DC Selco, DC DHA Selco and DC Super Selco) were used to enrich Artemia nauplii fed to seahorse, Hippocampus sp. fry. The emulsions varied in their n-3 highly unsaturated fatty acid (HUFA) composition. Total n-3 HUFA content ranged from 200 to 450mgg-1 between the three emulsions while levels of eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) ranged between 47–220 and 80–190mgg-1, respectively. Survival and growth of seahorses at the end of the 30 day growth trial were greater in treatments receiving enriched Artemia. Seahorses receiving Artemia enriched with DC DHA Selco and DC Super Selco showed significantly (p < 0.05) greater mean survival (71.6 ± 6.0% and 78.3 ± 6.0%, respectively) than those receiving unenriched Artemia (48.3 ± 6.0%). Mean standard length was also significantly greater (p < 0.05) in fry fed DC DHA Selco and DC Super Selco enriched Artemia (20.2 ± 0.3 and 19.7 ± 0.3mm, respectively) compared to those fed unenriched Artemia (18.1 ± 0.3mm). The results show that dietary n-3 HUFA are essential for optimal growth and survival of Hippocampus sp. and, based on the fatty acid compositions of the enriched Artemia used in this study, indicate that the level of dietary DHA supporting optimal growth and survival is greater than 9.3mgDHAg-1 dry weight.  相似文献   

13.
Wild striped bass,Morone saxatilis, were collected from coastal waters and spawning areas to describe the endocrine correlates of oocyte development in non-captive, migratory fish. The fish were classified according to their most advanced oocytes. Serum levels of estradiol (E2), testosterone (T) and 17-20-dihydroxyprogesterone (DHP) were measured by radioimmunoassay (RIA). Females in the primary growth phase and early secondary growth phase (pre-vitellogenic) had low levels of plasma steroids, ovarian lipid content and gonadosomatic indices (GSIs). Significant increases in E2, T, ovarian lipid content and GSIs occurred during the vitellogenic phase. Maximum levels of all reproductive parameters were found in prespawning fish sampled in the Hudson River. Mean levels of E2, T, ovarian lipids and GSIs for these fish were 2.0±0.5 ng/ml, 3.0±0.3 ng/ml, 24±1 mg/g, and 5.6±0.3% (mean±SEM), respectively. In fish induced to spawn with human chorionic gonadotropin (HCG), DHP levels (1.9±0.4 ng/ml) were significantly elevated. Similar levels were found in two fish captured during the spawning season, suggesting that DHP may serve as the maturation-inducing steroid in this species.  相似文献   

14.
Quantification of proteases activities and their class structure have been studied in a cladoceran, Daphnia carinata. Protease activity ranged from 0.28 to 0.55 Unit mg−1 protein min−1 with an average value of 0.42±0.06 Unit mg−1 protein min−1. Chymotrypsin activity was more than twofold higher (0.49±0.09 Unit mg−1 protein min−1) than the trypsin activity (0.21±0.02 Unit mg−1 protein min−1). Protease activity and reduction of activity in bands of samples treated with specific inhibitors were documented in photometric assay and substrate SDS–PAGE. Proteinase activity against azocasein was inhibited (91.4±1.5%) with SBTI. PMSF reduced the enzyme activity by 53.1±6.5%, and the azocasein hydrolysis was reduced up to 64.6±3.8% by the specific inhibitor of trypsin, TLCK. In the present investigation, the molecular weight of various activity bands ranged from 16.3 to 51.1 kDa. The molecular weights of several protein bands are similar to protease activity zones. The knowledge of digestive enzyme profiles of fish food organisms generated in the present study may assist in the formulation of age-specific feed.  相似文献   

15.
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   

16.
This study describes changes in cathepsin L activity during early development in Eurasian perch (Perca fluviatilis) and determines to which extend a precocious proteolysis of the yolk reserves by activated cathepsin L could be involved in the seasonal decline of egg and larval quality. During embryogenesis and early larval stages, samples were collected for determination of fertilization and hatching rates, assays of cathepsin L activity, and evaluation of larval resistance to stress (osmotic shock and fasting). Cathepsin L activity increased significantly during the embryonic development, reaching a maximum of 177.1±12.3 nmol min–1mg fresh weight–1 on day 1 post-hatching and significantly decreasing on day 3. Activity increased significantly during the spawning season and individual variations between egg strands became very high during the second half of the spawning period. An inverse relationship was established between the cathepsin L activity in 7-day old eggs and the decrease of hatching rate. A negative exponential regression was calculated between the cathepsin L activity of eggs and the resistance of corresponding newly-hatched larvae to osmotic shock. Resistance to fasting was significantly reduced in larvae from eggs with cathepsin L activity higher or lower than 20 nmol min–1 mg–1 (lethal time50=4.7±0.7 and 3.5±0.4 days, respectively). The involvement of cathepsin L in the degradation of yolk reserves and its potential consequences on the quality of perch eggs and larvae are discussed in relation to results reported in salmonid fishes.  相似文献   

17.
The Australian freshwater fish, silver and golden perch, are increasingly being used for aquaculture. Addition of salt to water is commonly used in commercial aquaculture to reduce stress attributed to high ammonia concentrations. The activities in gill homogenates of ouabain-sensitive Na+/K+-ATPase and NEM-sensitive ATPases (as a measure of H+-ATPases) of silver and golden perch were measured after maintaining the fish in water containing different salt and ammonia concentrations. Six treatments were applied in a 2 × 3 factorial design: two salt treatments, low salt (LS) of 2.5 g l− 1 and high salt (HS) 5 g l− 1, and three ammonia treatments, no added ammonia (NA), low ammonia (LA), 3 mg total ammonia nitrogen (TAN) l− 1 and high ammonia (HA), 5 mg TAN l− 1. In both species, activity of Na+/K+-ATPase was lowest in fish kept in the LSNA treatment (7.4 ± 0.4 μmol Pi mg protein− 1 h− 1 for silver perch and 3.1 ± 0.6 for golden perch) and highest in the HSHA treatment (15.2 ± 1.0 μmol Pi mg− 1 protein h− 1 for silver and 8.4 ± 1.2 for golden perch). In both species there was a significant increase (P < 0.001) in Na+/K+-ATPase activity with increase in salt concentration and with an increase in ammonia concentrations. A significant interaction (P < 0.036) between salt and ammonia on Na+/K+-ATPase activity was observed in silver but not in golden perch. In contrast, the lowest activity for NEM-sensitive ATPase was observed in the HSNA treatment (1.0 ± 0.2 μmol Pi mg− 1 protein h− 1 for silver and 1.5 ± 0.4 for golden perch) and highest in LSHA treatment (2.9 ± 0.4 μmol Pi mg− 1 protein h− 1 for silver and 3.6 ± 1.2 for golden perch). In both species there was a significant decrease in NEM-sensitive ATPase activity with increase in salt concentration and an increase in activity with increase in ammonia (P < 0.003). In silver perch, a significant interaction between the treatments was observed (P < 0.02). The results suggest that in these species of freshwater fish the Na+/K+-ATPase has a role in salt and ammonia homeostasis and that the NEM-sensitive ATPases are more active in fish kept in water with a lower salt content. It is possible that the increase in ammonia resistance when salt is added to the environmental water in commercial aquaculture systems may be due to the effects of salt on gill Na+/K+-ATPase activity rather than the NEM-sensitive ATPases.  相似文献   

18.
Specific binding sites for testosterone have been detected in three compartments of olfactory tissue from brown and rainbow trout. Binding of3H-testosterone to the membrane fraction of olfactory tissue is of high affinity (Kd = 0.5–1.9 nM) and limited capacity (Nmax = 30–60 fmol mg+1 protein). Binding is reversible, and is eliminated by protease treatment. The membrane binding site exhibits a high degree of ligand specificity; 11β-hydroxytestosterone, 11-ketotestosterone, 17α-hydroxyprogesterone, 17α,20β-dihydroxy-4-pregnen-3-one, cortisol, and estradiol-17β all fail to displace testosterone at 20-fold excess while testosterone itself competes successfully. These attributes are consistent with the presence of specific steroid receptor proteins. Binding of testosterone within the cytosol is of moderate affinity (Kd = 9.0–23.0 nM) and high capacity (Nmax = 0.5–2.9 pmol mg+1 protein) and is more readily displaced by a number of steroid competitors than is the case for the membrane site. The rate of association and dissociation of testosterone from the cytosolic binding site is markedly more rapid than the equivalent processes in the membrane fraction. Binding of testosterone to the nuclear extract is of high affinity (Kd ∼3.0 nM) and limited capacity (Nmax ∼50 fmol mg+1 protein). There are no substantial differences between species or between sexes in the affinity or capacity of testosterone-binding sites in nuclear extract or membrane fraction. However, cytosolic testosterone-binding sites are three- to four-fold more abundant in rainbow trout than in brown trout, and female rainbow trout have more cytosolic binding sites than male rainbow trout, but a lower affinity for testosterone than male sites. Preliminary evidence supports the involvement of the membrane-associated testosterone-binding site in olfactory processes. Rainbow trout display an EOG response to testosterone at a concentration (≥ 10+9 M) which is consistent with the equilibrium dissociation constant (Kd) of the membrane-associated testosterone-binding site. Binding of3H-testosterone to the membrane-associated site shows a pH dependency which is comparable to the effects of pH on the EOG response to testosterone in intact fish. The attributes of the intracellular testosterone-binding sites are common to testosterone receptors in other fish tissues which are known androgen target tissues. This suggests that the development and/or function of salmonid olfactory tissue may be susceptible to influence by endogenous testosterone.  相似文献   

19.
Histidine decarboxylase (HDC) from Staphylococcus epidermidis TYH1, a halotolerant histamine-producing bacterium isolated from Japanese fermented fish-miso, was purified to homogeneity for the first time. The enzyme was purified 182-fold from cell-free extracts by ammonium sulfate precipitation, anion exchange chromatography and gel filtration chromatography. The N-terminal amino acid sequences of two polypeptide chains of 27–30 and 7–9 kDa were highly homologous with those of α- and β-chains of other staphylococcal HDCs. The optimum pH and temperature for the enzyme were 6.0 and 60 °C, respectively. This enzyme did not decarboxylate lysine, arginine, tyrosine, tryptophan or ornithine. The enzyme activity decreased with the addition of NaCl. At pH 4.8, the V max and K m values were 45.5 μmol histamine min?1 mg?1 and 1.10 mmol/L, respectively. Moreover, this enzyme was resistant to heat treatment (80 °C for 15 min) and was stable upon freezing at ?30 °C for 7 days. The very similar physiological properties of this enzyme and the almost identical N-terminal amino acid sequence to that of the HDC from S. capitis indicated that this enzyme may be evolutionally highly conserved in the genus Staphylococcus. The biophysical properties of staphylococcal HDC were elucidated using native purified enzyme.  相似文献   

20.
This investigation examines the influence of implants containing 11-ketotestosterone (11KT), 17-estradiol (E2), and 3,5,3-triiodo-l-thyronine (T3) on astaxanthin metabolism in sexually immature individually tagged Arctic charr. The fish (initial average weight 427 g) were maintained in freshwater for 40 days, and weekly implanted intraperitoneally with oil-based injections containing either 11 KT, E2 or T3 at levels of 0.1, 1.0 and 0.1 mg (100 g body weight (BW))–1, respectively. The control fish were given the oil medium alone (0.2 ml 100 g BW–1). The diet contained ca. 50 mg astaxanthin kg–1. Carotenoid composition was monitored in plasma, fillet, liver and skin, and 11 KT, E2 and testosterone (T) levels in plasma. All hormone treatments reduced plasma T compared to the control. E2-treated fish had a higher (p<0.05) hepatosomatic index (HSI) than the other treatments. Hormone treatment did not influence gonadosomatic index (GSI). T3 administration induced a silvery skin appearance. The fillet and plasma carotenoid content decreased during the experiment. 11 KT implantation reduced astaxanthin and idoxanthin concentrations of plasma and fillets, and increased the amount in liver and skin, compared to the other treatments. The relative proportion of astaxanthin to idoxanthin was higher in the control fish and T3 implanted fish, than in fish implanted with 11 KT or E2 (p<0.05). Fish treated with E2 had the highest skin carotenoid concentration. Male fish had significantly higher carotenoid content in plasma, fillet and skin than female fish. This study reveals that sex hormones affect carotenoid metabolism and partitioning among body compartments of Arctic charr, effects differently displayed by the sexes.  相似文献   

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