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1.
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2.
The structural and oxygen binding properties of Acipenser naccarii blood have been investigated. The electrophoretic analysis of the hemolysate of this sturgeon showed the presence of two hemoglobin components, each with a considerable globin multiplicity. Constituent globin chains were analyzed by urea-Triton acid polyacrylamide gel electrophoresis and isolated by high performance liquid chromatography. N-terminal amino acid sequence analysis revealed the presence of a N-terminal proline in two of the three -chains present in the globin pattern, and the presence of a histidine residue in 2position. Oxygen equilibria reveal a very low sensitivity of the individual hemoglobins to chloride ions and temperature; however, in the presence of organic phosphates the oxygen affinity of the hemoglobin components decreases strongly. In particular, when Guanosine-5-triphosphate (GTP) is added, the reduction of the oxygen affinity, at pH 7.4 and 20 °C, is 60% and 50%, respectively, for HbI (anodic component) and HbII (cathodic component). As the effect of protons is concerned, the small Root effect shown by total hemolysate at physiological conditions, seems to be due mainly to the cathodic component. On the whole, the functional properties shown by sturgeon hemoglobin components seem to be related to the particular physiological needs dictated by the environmental characteristics.  相似文献   

3.
The nutritional effect of vitamin E in dietsfor Litopenaeus vannamei postlarve (PL19)was investigated. Four formulated diets withdifferent combinations of -tocopherylacetate (-TA), ascorbic acid (AA) andhighly unsaturated fatty acids (HUFA) weretested, using four replicates.No significant differences in survival wereobserved among treatments after 34 days offeeding. However, shrimp fed with a dietcontaining 2% fish oil (low n-3 HUFA content),200 mg.kg–1 -TA and100 mg.kg–1 AA (diet H/E/C) showedsignificantly better growth than those fed adiet supplemented with 5% fish oil (high n-3HUFA content), 200 mg.kg–1 -TA and100 mg.kg–1 AA (diet H+/E/C). Shrimp fedwith a diet containing 5% fish oil,900 mg.kg–1 -TA and100 mg.kg–1 AA (diet H+/E+/C) showed a significantly higher tissue level of n-6 PUFAthan postlarvae fed diet H+/E/C. No definiteconclusion could be drawn about a possibleinteraction between -TA and AA, since acomparison of the diet containing 5% fish oil,200 mg.kg–1 -TA and700 mg.kg–1 AA (H+/E+/C+) and the dietH+/E/C did not show any significant differencesin any of the measured parameters. Theantioxidative status of the shrimp tissue(measured by means of the thiobarbituric acid(TBA) assay and expressed as nM malonaldehyde(MA) per gramme dry weight) was equal for alltreatments. Nevertheless, there was a slightlylower MA value with the diet H+/E/C+,indicating that AA may be an effectiveantioxidant in the aqueous phase and at thewater/lipid interface of the tissue. The tissuelevels of -T and AA were highlydependent on the amounts in diets and nocorrelation between -T and AAincorporation could be observed.  相似文献   

4.
Changes in 17-estradiol (E2), estrone (E1), testosterone (T), 11-ketotestosterone (11KT), and 17,20-dihydroxy-4-pregnen-3-one (17,20-P) levels were correlated to changes in gonadosomatic index (GSI), vitellogenin concentration (Vg), ovarian and testicular histology during the annual reproductive cycle of the red porgy, Pagrus pagrus. The production of E2, E1, T and 17,20-P was confirmed by analysis of the steroidogenic activity of ovaries. In females, the average concentration of E2 was lower than 2 ng ml–1. E2 values first increased significantly at the stage of endogenous vitellogenesis and remained high during exogenous vitellogenesis. E1 levels were lower values than E2 (less than 300 pg ml–1), but they increased at the beginning of exogenous vitellogenesis. Estrogens concentrations followed similar pattern to Vg and were significantly correlated. Mean levels of T were mostly lower than 1 ng ml–1. They followed a pattern similar to that of E2 except for a further increase observed at the stage of final maturation. T and E2 levels were significantly correlated. The concentration of 11KT did not change significantly. The levels of 17,20-P ranged between 0.22 and 1.22 ng ml–1 but changes were not related to gametogenesis. In males, the concentrations of T and 11KT fluctuated significantly during the sexual maturity stages, showing a similar pattern and were significantly correlated to GSI changes. T levels increased during spermiogenesis and spermiation stages to reach about 3 ng ml–1. 11KT levels stayed about half those of T. The levels of estrogens showed no significant changes. Level of 17,20-P showed no significant variation related to male maturity. Results are discussed in relation to changes in plasma steroid levels during gametogenesis of other multiple spawner species.  相似文献   

5.
The female bambooleaf wrasse, Pseudolabrus japonicus, spawns daily during the spawning season, and exhibits a diurnal rhythm of ovarian development. In the present study, we have investigated: (1) circulating levels of 17a, 20-dihydroxy-4-pregnen- 17,20-P) and 17,20,21-trihydroxy-4-pregnen-3-one (20-S) in females sampled at different times of the day during spawning season in captivity, and (2) in vitro production of 17,20-P and 20-S by follicle-enclosed oocytes at seven different develo tal stages. In addition, we developed a microtiter plate enzyme-linked immunosorbent assay (ELISA) for 17,20-P. Serum levels of 17,20-P and 20-S showed similar diurnal changes; substantial increases in these levels occurred around the time of germinal vesicle breakdown (GVBD). In vitro experiments showed that massive production of 17,20-P and 20-S occurred in follicles collected just before or during GVBD. Further, acute decreases in 17,20-P and 20-S production were found in the ovarian follicles just prior to ovulation, suggesting inactivation of the maturation-inducing hormone (MIH). These results, taken together with our previous data on the occurrence of GVBD in vitro, suggest a role for both 17,20-P and 20b-S as MIHs in the bambooleaf wrasse.  相似文献   

6.
This study was conducted to evaluate theeffect of plant proteinbased diets on gonadaldevelopment and plasma 17 -estradiol (E2) levelin female Nile tilapia Oreochromis niloticus.Fish with a mean body weight of 6.7 (0.1) g were fedfour different diets with the same digestible protein(DP) and digestible energy (DE) containing gradedlevels of a mixture of plant ingredients as partial ortotal replacement of fish meal protein for 20 weeks.The control diet (D0) was based on fish meal, twodiets containing 33% (D33) and 66% (D66) of plantprotein, and one diet containing only plant protein(D100). Fish were sampled at 12 and 20 weeks. Nosignificant differences were found in different stagesof oocyte development and plasma E2 levels betweentilapia fed diets D0 and D100 at 12 weeks. Eight weekslater tilapia fed diet D0 showed a higher (P < 0.05)level of E2 than the D100 group. This difference andthe reduced proportion of vitellogenic and matureoocytes demonstrated that diets containing only plantprotein are less efficient in terms of tilapia growthand consequently ovarian development.  相似文献   

7.
莫荷罗非鱼“广福1号”与其亲本间DNA甲基化的差异分析   总被引：1，自引：1，他引：0  
为了探讨罗非鱼杂种优势形成过程中基因组DNA甲基化模式的变化,运用甲基化敏感扩增多态技术(methylation sensitive amplified polymorphism,MSAP)分析橙色莫桑比克罗非鱼、荷那龙罗非鱼及其杂交种莫荷罗非鱼"广福1号"的皮肤、肌肉和鳃等11种组织的DNA甲基化水平差异。采用16对引物进行选择性扩增,经聚丙烯酰胺凝胶电泳带型分析结果显示:莫荷罗非鱼"广福1号"及其亲本在不同组织间的甲基化水平存在组织特异性,相同组织不同亲、子代罗非鱼间的甲基化程度亦不同;橙色莫桑比克罗非鱼、荷那龙罗非鱼以及"广福1号"各组织平均总甲基化水平分别为32.21%、38.03%和29.77%,莫荷罗非鱼"广福1号"的甲基化水平低于双亲;与亲本相比,莫荷罗非鱼"广福1号"的甲基化模式除了大多数保持稳定遗传(A、B、C类型分别为19.67%、29.99%、25.42%)之外,24.92%胞嘧啶位点发生去甲基化和超甲基化,且去甲基化位点(E类型,15.73%)多于超甲基化位点(D类型,9.19%)。研究表明,莫荷罗非鱼"广福1号"基因组DNA的低甲基化特征和甲基化模式的重调可能与耐盐杂种优势有关。  相似文献   

8.
The role of calcium influx through voltage-dependent calcium ion channels in the exocytotic response of single isolated pars intermedia cells from the teleost tilapia (Oreochromis mossambicus) was investigated by means of the whole-cell patch-clamp technique and high resolution electrical measurements. Calcium currents differed from barium currents in several ways: the peak ICa was smaller, the current-voltage relationship for calcium attained its maximum at +10 mV instead of 0 mV, and the inward calcium current inactivated more rapidly. Electron micrographs showed that pars intermedia cells possess dense, encored vesicles with an average diameter of 140 nm. Influx of calcium resulted in an increased cell membrane capacitance (Cm) after the depolarizing period, indicating a fast exocytotic response. Comparison with late recordings revealed the presence of a transient in Cm, presumably attributable to movement of ion channel gates. The average increase in Cm was 13.4 fF, suggesting the fusion of at least 23 vesicles with the plasma membrane during the depolarizing pulse. In contrast to calcium, barium did not support significant exocytosis. We conclude that calcium entry through voltage-dependent calcium channels rapidly leads to the exocytosis of secretory vesicles from tilapia pars intermedia cells.  相似文献   

9.
This study used random amplified polymorphic DNA (RAPD) fingerprinting for estimating genetic variation and species differentiation in three species of tilapia. A 16-mer random primer generated RAPD markers ranging from 250 to 2400 base pairs (bp). Genetic similarity estimates obtained by pairwise comparisons based on the method of Nei and Li (1979) indicated high genetic similarity (mean genetic similarity (± sd), 0.73 (± 0.15) for Nile tilapia; 0.78 (± 0.12) for Mozambique tilapia; and 0.87 (± 0.07) for Aureus tilapia) within each of the tilapia species. The average interspecies genetic similarities obtained among the three species were 0.59 (± 0.07) for Mozambique/Nile tilapia, 0.46 (± 0.09) for Aureus/Nile tilapia and 0.38 (± 0.07) for Aureus/Mozambique tilapia pair. DNA profiles generated in each species of tilapia were unique. A total of 13 RAPD markers differentiating the three species of tilapia were detected. Our study presented RAPD markers as a new class of useful genetic markers for assessment of genetic diversity and species differentiation in tilapia.  相似文献   

10.
Two experiments, oral carbohydrate administration (Experiment 1) and vein glucose injection (Experiment 2), were conducted to gain more insight into the ability of hybrid tilapia, Oreochromis niloticus × O. aureus, to utilize different carbohydrates and to establish the kidney threshold for urinary glucose excretion. In Experiment 1, both glucose and starch were administered orally after the tilapia were fasted for 24 h. Plasma and urine were sampled from the fish at selected time intervals from 1 to 24 h thereafter. Higher (p<0.05) plasma and urine glucose concentrations were found in fish fed on glucose than in fish fed on starch. The concentration of plasma glucose of tilapia peaked at 3 h (25.45 mM for glucose; 8.24 mM for starch) after the oral ingestion of both carbohydrates. Maximum urinary glucose concentrations (48 mM for glucose; 10 mM for starch) in fish fed glucose and starch were at 3 and 4 h post administration. In Experiment 2, five concentrations (0, 0.08, 0.12, 0.16 and 0.24 g glucose ml–1) of glucose solution were injected into the caudal vein of the tilapia. Urine were sampled from the fish at 30-min time intervals from 0 to 6 h after the injection. Blood was sampled at 1 h after the injection. Higher urinary glucose concentrations were observed in fish injected with 0.12 g glucose ml–1. When the urinary glucose concentrations in fish injected with the various glucose concentrations were plotted against the plasma glucose concentrations of the fish 1 h after injection, the kidney threshold for urinary glucose excretion in tilapia appeared to be about 6 mM.  相似文献   

11.
Five experimental diets containing increasing proportions of cottonseedmeal (CSM) protein (0, 25, 50, 75 and 100%; diets 1 to 5, respectively) toreplace fish meal (FM) protein were formulated for intensive culture of tilapiaOreochromis sp. Each diet was fed to three replicategroupsof fish (mean weight ± SE = 11.3 ± 3.9 g) in30L aquaria connected as a closed recirculating-water system andmaintained at 27 ± 1 °C. Fish were fed three times adayby hand at a rate of 3% of body weight during four weeks, after which thefeeding rate was gradually decreased to reach 1.5% at 16 weeks. Thesubstitutionof 75 and 100% of FM proteins by CSM proteins resulted in significantly lowerbody weights in both sexes. In both sexes, gonadosomatic indexes and plasmaconcentrations of sex steroids (testosterone, 11-ketotestoterone,estradiol-17 and 17,20-dihydroxy-4-pregnen-3-one) were notsignificantly different among dietary treatments. The concentration ofgossypol,an antifertility agent contained in CSM, was measured in reproductive tissues.The total gossypol concentration in the testis was consistently lower than thatmeasured in the ovaries of the same group. Moreover, in both sexes, theconcentration of the (+)isomer of gossypol was always higher than that of(–)isomer. The total gossypol concentrations in testes increasedsignificantly with the increase of CSM in the diet. The highest levels of the(+)isomer (7.64 ± 0.62 g g–1)were found in the testes of fish fed diet 4, whereas the (–)isomerreached its highest values in the testes of fish fed diet 5. The highest levelsof both enantiomers of gossypol were found in the ovaries of fish fed diet 4(14.2 ± 2.7 and 5.6 ± 1.5 g g–1for (+) and (–)isomers, respectively). In both sexes, thehistological analysis of the gonads did not reveal differences among the fishfed different levels of CSM. Although CSM at any levels did not affect thereproductive parameters examined in this study, it cannot be used to substitutemore than 50% of FM since at higher levels growth of tilapia was compromised.  相似文献   

12.
Roach ovaries converted 17-hydroxyprogesterone to 17,20-dihydroxy-4-pregnen-3-one (17,20P) and to glucuronides of testosterone and 17,20P. Small amounts of 5-pregnane-3- and -3, 17, 20-triols, 7-hydroxy-5-reduced metabolites and 17,20-dihydroxy-4-pregnen-3-one (17,20P) were also formed. Rudd ovaries converted this substrate mainly to 17,20P, 5-pregnane-3- and -3,17,20-triols, 17,20-dihydroxy-5-pregnan-3-one and testosterone glucuronide. The main metabolites of progesterone with both species were 17,20P, 5-pregnane-3,17,20-triol and 7-hydroxy-5-reduced steroids. Rudd ovaries formed, in addition, 17,20-dihydroxy-5-pregnan-3-one from progesterone. The pattern of metabolites was markedly altered when the concentration of substrate was increased from 42ng to 1 µg or 100 µg. At the highest concentration, glucuronides and polar steroids were not detectable, while at low concentrations they accounted for over 50% of the metabolites. 20-Hydroxysteroid dehydrogenase was shown to have a very high capacity, producing 21–47 µg 17,20P from 100 µg 17-hydroxyprogesterone substrate with 200 mg ovarian tissue in 5h.  相似文献   

13.
Samples of fertilized dentex, Dentex dentex, eggs and larvae fed enriched rotifers and Artemia according to standard hatchery procedures were analysed for free, total amino acid and fatty acid contents. Egg free amino acids (53 nmol ind-1) and total lipids (13.5 (g ind-1) levels were considerably reduced in the newly hatched larvae (6.0 nmol and 5.7 (g ind-1 respectively) while the amount of -3 PUFA (Polyunsaturated fatty acid) and 20:4-6 remained unchanged suggesting their possible role as essential fatty acids (EFA) in this species. In general, the changes in pattern of the major biochemical components during early development of dentex was similar to that outlined for other Sparidae species investigated to date. The essential amino acid profile of enriched live food did not show major imbalances when compared to that of dentex larvae. In contrast, the level of docosahexaenoic acid (22:63, DHA) and the DHA:EPA (eicosapentaenoic acid, 20:5-3) ratio in larvae given rotifers and Artemia were significantly lower relative to the corresponding values in the unfed larvae.  相似文献   

14.
Four African wild strains (Egypt, Ghana, Senegal and Kenya) and four established Asian farmed strains of Nile tilapia, Oreochromis niloticus (popularly known in the Philippines as Taiwan, Thailand, Singapore and Israel) were analysed electrophoretically at 30 protein loci to estimate genetic differences among the strains. All strains shared alleles at 14 monomorphic and 16 variable loci. Among the African strains, characteristic allele frequency differences were observed at AAT-1 * 46 for Ghana and Senegal, ADH * 83 for Kenya, ADH * 120 for Senegal, G3PDH-2 * 300 for Egypt, IDDH * 67 for Senegal, sMDH-1 * 120 for Kenya and SOD * 150 for Senegal. Genetic distance values among the strains revealed a clustering of the farmed strains with Egypt and Ghana O. niloticus, a slight separation of the Senegal strain and a larger separation of the Kenya strain. This profile may reflect the origins of the few founder populations of this species previously introduced to Asia. It also confirms the wider genetic divergence of the Kenya strain (O. niloticus vulcani) from the others studied here, which are all O. n. niloticus. Observed heterozygosities of the strains ranged from 0.026 to 0.071, with the African wild strains the lower values (mean Ho = 0.036) and the farmed strains the higher ones (mean Ho = 0.056). The implications of these results to the ongoing tilapia genetic improvement programme in the Philippines are discussed.  相似文献   

15.
Distribution and function of hepatic - and -adrenoceptors were examined in rainbow trout (Oncorhynchus mykiss) injected with slow release hydrogenated coconut oil implants alone (sham) or containing cortisol. - and -Adrenoceptors were assayed on purified hepatic membranes 10–14 days post-implantation using 3H-prazosin () and 3H-CGP (). At 10–14 days, plasma cortisol values were significantly elevated to approximately 220 compared with 35.0 ng ml-1 in cortisol implanted vs. sham trout. No significant differences were found between any of the experimental groups for either the affinity (Kd) or maximal number of binding sites (Bmax) for either receptor type. Epinephrine significantly stimulated glucose release from hepatocytes isolated from sham injected trout, but not from cortisol-treated fish. Epinephrine-induced glucose release was blocked by both - and -antagonists. These studies do not support the hypothesis that rainbow trout exposed to chronic cortisol alter properties of hepatic adrenoceptors.  相似文献   

16.
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   

17.
Pituitaries of various teleosts belonging to 25 orders were immunostained with antisera raised against synthetic fragment peptides corresponding to conservative regions of gonadotropin subunits (mummichog FSH 50-60 and mummichog LH 91-106). Both immunoreactive FSH cells and immunoreactive LH cells were successfully identified in the fishes of almost every order of the superorder Acanthopterygii and several species of the superorders Paracanthopterygii and Polymixiomorpha, such as mullet, alfonsino, flyingfish, mackerel, flounder, cod, beardfish, etc. These antisera are therefore considered as universal antisera for immunocytochemical application to acanthopterygian fishes. Extensive diversity in the abundance of the FSH cells and the LH cells among species was noted even in fishes with similar gonadal stages, indicating the possibility that the respective roles of FSH and LH may vary considerably among species in advanced teleosts.Evident but generally weak immunoreactivities to anti-mummichog LH 91-106 were observed in the fishes of the superorder Cyclosquamata; and slight or weak immunoreactivities to the antiserum were observed in the fishes of several more primitive taxa (superorder Stenopterygii, Protacanthopterygii, Ostariophysi, subdivision Clupeomorpha, and subdivision Elopomorpha). No immunoreactivity to anti-mummichog FSH 50-60 was observed in these fishes. These results are consistent with the phylogenetic status of the fishes and the degree of conservativeness in the amino acid sequences of the antigen regions.  相似文献   

18.
Recent DNA comparisons confirm that the endangeredLake Eacham rainbow fish (Melanotaenia eachamensis Allen andCross, 1982) is a separate species, distinct from othermelanotaeniids in the area. Although the large populationformerly in Lake Eacham has not been re-established, other smallnatural populations of M. eachamensis have been identified fromLake Euramoo and Dirran Creek.Commencing with 23 captive individuals (3 14 6 I), representing three original stocks, Project Eachamensis (initiated by the Australia New Guinea Fishes Association in July 1987) has stimulated and actively encouraged the establishment of new captive populations. Approximately 15 captive breeding populations are now reported in eastern Australia -- numbers of individuals are estimated to exceed 1000; additional Bowman and Tappin populations are being maintained in North America and Europe.A recent review of this conservation programme (involving individual aquarists, hobbyist organizations and scientists as well as the aquarium industry and government) has now recommended: an alteration to the conservati on status category from endangered to vulnerable; maintenance of existing captive populations; formulation of a comprehensive recovery plan with the long-term objective of re-establishing and increasing natural populations; and encouraging public education on the risks of translocation.  相似文献   

19.
The purpose of this work was to study the turnover of a, andtocopherol (TOH) in Atlantic salmon (Salmo salar, L.). Fish induplicate tanks were fed a diet containing 150 mg kg-1-TOH and 100 mg kg-1 each of and TOHadded as tocopheryl acetates. After fillet TOH concentrations had adjustedto the dietary supplementation levels, samples were taken from fish that hadbeen deprived of feed for 100 h, and from fish that had been fed regularlyuntil sampling. The retained levels of tocopherols in plasma correspondedgrossly with their biological activities, as found in experiments withmammals (::100: 20:3). The plasma concentrationsof -, and TOH amounted to 65, 44 and 15%,respectively, in unfed compared to fed fish. Very low density lipoprotein(VLDL), appeared to contain a greater fraction of plasma -TOH than ofplasma TOH. The mitochondrial fraction of liver, but not that of darkmuscle, was highly enriched in -TOH, and less in andTOH. The concentration ratios in liver and bile indicate that, and to some extent, TOH are excreted in the bile at a higherrate than -TOH. The data fit the hypothesis that Atlantic salmonliver contains a tocopherol binding protein with higher affinity for-TOH than for the other tocopherol homologues. This appears toprevent excretion of -TOH in the bile, and stimulate incorporation of-TOH in VLDL for subsequent secretion into the blood stream. As aconsequence, -TOH is retained in the body to a greater extent than and -TOH.  相似文献   

20.
We have isolated a cDNA encoding the taurine transporter from a tilapia (Oreochromis mossambicus) gill cDNA library. Transient expression of the cDNA in COS-7 cell indicates that the clone encodes a Na+- and Cl–-dependent and -amino acid-specific taurine transporter. By the transfer of tilapia cultured in freshwater to 70% artificial seawater, plasma osmolality increased by up to 100–135 mOsm/kgH2O along with the marked increase in the taurine transporter mRNA level in all the tissues examined i.e., kidney, stomach, intestine, gill, eye, liver, fin, muscle and brain. In most tissues, time-dependent change in the taurine transporter mRNA level corresponds to that in plasma osmolality. However, fin showed an acute and muscle showed a delayed increase in taurine transporter mRNA compared to changes in plasma osmolality. The taurine transporter mRNA level in tilapia embryos also increased after transfer from freshwater to 100% artificial seawater. Increase in taurine transporter expression leads to the activation of cellular uptake of taurine from plasma and the accumulation of taurine in the cell. Thus the results in the present study suggest that taurine plays an important role as an osmolyte in the ubiquitous tissues of tilapia during high-salinity adaptation.  相似文献   

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