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1.
Spermatozoa and seminal plasma obtained from rainbow trout and whitefish were analyzed in respect to their aspartate aminotransferase (AspAT) and alkaline phosphatase activities. In particular, the experiments characterized AspAT optimum pH, optimization of assay conditions and action of coenzyme, pyridoxal 5-phosphate (vitamin B6). The effect of short-term semen storage at 0°C on biochemical indicators and fertilization rate was examined in both species. The concentrations of reduced and oxidized ascorbic acid in seminal plasma of both species were several folds higher than in spermatozoa and blood plasma of fish. Highly significant correlations were found for both species between AspAT activity (sperm or seminal plasma) and fertilization rate (% of eyed-stage or hatched embryos). For rainbow trout, highly significant correlations were found between sperm concentration, motility and fertilization rate. These results suggest that several biochemical indicators of seminal plasma can be used as measures of sperm quality of fish. Some common biochemical parameters for fish and mammal's semen provide evidence for using fish sperm as a model in biomedical research.  相似文献   

2.
During the reproductive season, rainbow trout spermatozoa are stored in the sperm ducts for several months. There is no sperm production at this time since spermatogenesis is completed before spawning. To leam more about characteristics of semen during such a long storage, we analyzed changes in protein concentrations, anti-proteinase activity in seminal plasma and sperm aspartate aminotransferase activity during an extended reproductive period during which fish were fed diets supplemented with various ascorbic acid concentrations. Seminal plasma protein concentration and anti-proteinase activity declined toward the end of the reproductive season. These phenomena may be related to oncoming proteolytic events leading to degradation of the sperm. Protein concentrations and anti-proteinase activities were strongly correlated within groups of different ascorbic acid supplementations and several sampling dates (r=0.6–0.9 in most cases, p<0.05). Ascorbic acid deficiency resulted in a decrease in both parameter levels as compared to levels in groups with vitamin C supplement (p<0.08). Deficiency also resulted in lower stimulation of aspartate aminotransferase by an exogenous pyridoxal 5-phosphate in comparison to fish fed vitamin C-supplemented diets (p<0.05). These results support earlier studies suggesting a protective role of ascorbic acid toward maintaining sperm quality.  相似文献   

3.
Determination of semen quality is necessary to understand the basic biochemical processes occurring during motility of sperm and during fertilization to evaluate the reproductive ability of different fish species and to create an optimal environment for storage of spermatozoa; in this regard less information is available for Acipenseridae compared with Cyprinidae and Salmonidae. The aim of the present study is to determine chemical composition and osmolality of seminal fluid and their relationship with sperm motility in Acipenser persicus. The results obtained show that sodium (Na+), chloride (Cl?) and potassium (K+) were predominant ions in the seminal plasma and the average of osmolality of seminal plasma was 82.56 mOsm kg?1. The higher chemical contents and osmolality compared with other sturgeon species reveal species‐specific characteristics and high secretory activity of spermatic duct in A. persicus. Significant positive correlations were observed between osmolality‐Cl?, Na+‐osmolality and Na+–Cl? (P<0.05, P<0.001 and P<0.05 respectively). But statistically significant correlation was not observed between seminal plasma parameters and sperm motility. Probably, the Na+ and Cl? are the main electrolytes playing a major role in maintaining the osmolality of the seminal plasma and the viability of the spermatozoa in vivo.  相似文献   

4.
Sperm physiology, in vivo artificial insemination and spawning of the ocean pout (Macrozoarces americanus L.), a marine bottom fish, were studied. Milt was collected from the reproductive tract of mature males by suction using a catheter. The uncontaminated milt, having a very low sperm concentration, contains highly motile spermatozoa and sperm motility was retained in vitro at 4 °C for at least 24 h in both seminal plasma and ovarian slime collected from the oviduct of pre-spawning females. Instead of activating sperm, dilution in sea water instantly immobilized the spermatozoa of ocean pout. Osmolarity and pH of ocean pout seminal plasma were in the ranges 365–406 mOsM and 7.2–7.5, respectively. A study of the ionic composition of ocean pout seminal plasma demonstrated the presence of various ions including Na+, K+, Ca2+, Mg2+, and Cl−, with a remarkably lower K+ concentration compared to that from other fish species. Since injections of milt containing motile sperm into the ovaries of pre-spawning females, which spawned in the absence of males, yielded fertilized ocean pout eggs, it is concluded that the ocean pout exhibits internal fertilization. The larvae hatched after 3 months of egg incubation in ambient sea water (9–10 °C). With proper timing of in vivo artificial insemination of mature females, fertilized ocean pout eggs can be obtained from fish reared in captivity.  相似文献   

5.
This mini-review focuses on changes in ATP and creatine phosphate concentrations in fish sperm under storage conditions. The storage of catfish sperm at 4°C leads to ATP depletion and decreased sperm motility. The rate of intracellular ATP depletion can be diminished through the addition of energetic substrates to the sperm storage medium, with lactate + pyruvate being the most efficient substrates for maintaining ATP concentrations in catfish sperm. The decrease in ATP concentration is closely associated with increases in AMP and hypoxanthine content. In contrast to catfish sperm, carp sperm is able to maintain intracellular ATP concentration close to the physiological level during storage. Collectively, these results suggest that fish species differ in terms of the energy metabolism of their spermatozoa and that the semen storage medium must be carefully selected for a particular fish species so as to maintain the ATP concentration and adenylate energy charge close to physiological values as long as possible.  相似文献   

6.
Sperm cryopreservation has led to transcendental changes in the reproductive biotechnology of both mammals and fish, and is a basic tool for animal improvement. However, these protocols generate damage to cell structure and physiology, altering sperm function as a result of cryoinjuries during freezing and thawing. This review is a compilation of the techniques developed and standardised for assessing sperm function in cryopreserved fish semen. Recent studies have analysed sperm function objectively, applying cellular and molecular techniques to characterise cryodamage. The Computer Assisted Sperm Analysis system has facilitated the assessment of motility, while electron microscopy (SEM and TEM) and cryo‐microscopy have made it possible to study sperm morphology and ultra‐structure. The effects of cryodamage on nuclear DNA have also been analysed using various methods, including the comet Fluorescence in situ Hybridization test, TUNEL, Sperm Chromatin Structure Assay, specific DNA sequences using RT‐PCR and specific genes by qPCR. The latter technique is used to study the mitochondrial genome (mtDNA), together with some candidate genes which are associated with bioenergy activity and sperm motility. Other parameters assessed are mitochondrial membrane potential and ATP content using high performance liquid chromatography, nuclear magnetic resonance spectrometry and cell respiration. All this information makes it possible to establish study and assessment criteria for cryopreserved fish spermatozoa. This work focuses on the use of technologies to study of quality of fish spermatozoa during cryopreservation.  相似文献   

7.
Understanding the effects of environmental factors in sperm qualities will be helpful in the development of optimal artificial reproduction methods and contributes towards the knowledge base of better short‐ and long‐term fish semen preservation conditions The objectives of this study were to determine properties and activities of wild‐caught striped jewfish Stereolepis doederleini sperm contaminated with blood or seawater and compare them with data reported in the literature on other freshwater and marine fish species, for effective short‐ and long‐term storage of fish semen. Overall, we observed that the sodium, chloride, glucose, total protein concentrations of normal sperm were not significantly different from blood‐ or seawater‐contaminated sperm. The salinity and osmolality concentration of sperm contaminated with blood were lower than sperm contaminated with seawater and were not significantly different from normal sperm. In addition, the spermatozoa motility (SM) and duration of spermatozoa motility (DSM) in blood‐contaminated sperm were higher than seawater‐contaminated sperm and also not significantly different from normal sperm. The best condition for SM and DSM in normal sperm was dilution rate of 1:50. Sperm was immotile in distilled water, and cationic factors were shown to stimulate the initiation of spermatozoa activation. The maximum SM and DSM were observed in solution containing 0.4 M NaCl, 0.6 M KCl, 0.6 M CaCl2 and 0.4 M MgCl2. This study provides some basic and important knowledge about striped jewfish sperm sensitivity to a cationic condition. In this regard, Na+ is the major inhibitory factor of spermatozoa motility in this fish species.  相似文献   

8.
The Ionic and Osmotic Factors Controlling Motility of Fish Spermatozoa   总被引：4，自引：0，他引：4  
This review presents actual knowledge about energetic, ionic, osmotic and gaseous control of fish sperm motility and its duration. Right after they are activated, fish spermatozoa of most species swim for a short period of time, in the range of one to several minutes. What determines the activation process? Is it due to the new ionic, gaseous and/or osmotic environment? Why is the duration of motility so short? Is it resulting from a fast exhaustion of energy stores (ATP, ADP, AMP, PCr) combined with the above-mentioned ionic/osmotic stress leading to morphological alterations? The motility criteria (flagellar beat frequency, head displacement velocity, flagellar waves morphology, etc.) used to characterize fish sperm movement and sperm flagella will be described. Most parameters change very rapidly during the brief motility period of fish sperm. Then will be considered the main environmental factors, ionic and/or osmotic signals, responsible of the activation of fish sperm motility. Then the metabolic compounds involved in cell energetics will be considered as their concentrations also rapidly change during the motility phase. An additional feature will then be discussed concerning the mechanisms by which fish sperm cell can be revived for a second motility round at the end of the first motility period. A model is proposed to explain how external osmolarity can control internal ionic composition, the latter being the key factor controlling flagellar activity.  相似文献   

9.
Oocyte growth in most oviparous vertebrates including fish is due to the formation of yolk, and eggshell proteins (zona radiata proteins). Zonagenesis leads to the formation of zona radiata proteins in oocytes, which play an important role during oogenesis, whereas vitellogenesis leads to the formation of yolk in oocytes through a series of events during which the yolk precursor protein vitellogenin (Vg) is synthesized and secreted from liver into blood from where it is sequestered into the developing oocytes and thereafter proteolytically cleaved to form yolk proteins (YPs) and finally deposited in the ooplasm. Much research has been done in many fish species with respect to the number and nature of Vg and YPs and their probable functions during fish reproduction. Recent findings of multiplicity of Vg molecules in fishes reject the earlier view of a single-Vg model and have led scientists to explore the functions of individual Vg and their YP derivatives, lipovitellin, phosvitin, and β′-component. Two distinct types of Vg or Vg genes, containing or encoding the three YPs, have been detected in many teleosts. A third unusual, incomplete, phosvitin-poor Vg has been described recently in many fishes. In comparison to much of the information on vitellogenesis in many fishes very little is known for Indian fishes. In India research has been done in a few species such as the catfish, Heteropneustes fossilis and Clarias batrachus, the murrel, Channa punctatus and the Indian major carps, Labeo rohita and Cirrhinus mrigala. Immunological and biochemical analyses suggest the occurrence of multiple forms of Vg and their YP derivatives. The synthesis and incorporation of Vg are regulated by gonadotropin (GTH) and estradiol-17β (E2). A differential role between estrone (E1) and estriol (E3) has been demonstrated for Vg synthesis. Enzyme-linked immunosorbent assays (ELISAs) for Vg have been developed to measure plasma Vg. Finally the different roles of Vg1 (HAI) and Vg2 (HAII) on vitellogenesis have been demonstrated. However, more research remains to be carried out in other fish species with respect to the number and nature of Vg and YPs and their genes in order to describe their reproductive functions.  相似文献   

10.
Oxidative stress is a possible source of spermatozoa function deterioration. Seminal fluid (SF) protects spermatozoa against reactive oxygen species (ROS) attack during development in testes and transit through the reproductive tract. Spermatozoa curvilinear velocity and percent of motile cells as well as changes in thiobarbituric acid-reactive substance (TBARS) content, superoxide dismutase, and catalase activity, and uric acid concentration in SF were evaluated in sterlet sperm collected from testes 24 h after hormone induction of spermiation and from Wolffian ducts at 12, 24, 36, and 60 h after hormone injection (HI). While testicular spermatozoa motility was not initiated in activating medium, Wolffian duct sperm showed low motility at 12 h, significant increase at 24 and 36 h, and decrease at 60 h. Testicular SF was characterized by the highest level of TBARS and activity of studied enzymes compared with SF from Wolffian duct sperm at 24 h post-HI. In fluid from Wolffian duct sperm, a significant increase in TBARS content was shown at 36–60 h post-HI. In contrast to testicular SF, in SF from Wolffian duct sperm, this increase was not counterbalanced by changes in the studied variables of antioxidant system. This may be the source of the observed decrease in spermatozoa motility parameters 60 h post-HI. The results may confirm a dual role of ROS in fish sperm physiology. The data with respect to decrease in sturgeon spermatozoa motility parameters at 60 h post-HI should be taken into account in artificial sturgeon propagation.  相似文献   

11.
Sixteen monthly collections of adult male Gambusia holbrooki (eastern mosquitofish) were obtained from two lakes in central Florida, USA. Lake Woodruff and Lake Apopka are 36 miles apart, but differ in several environmental parameters. Compared with Lake Woodruff, Lake Apopka is warmer, more shallow in sampling areas (particularly during drought conditions; approximately 15–90 cm in Lake Apopka versus 60–120 cm in Lake Woodruff), more turbid, and more heavily contaminated with nutrients and industrial and agricultural chemicals. Here, we present detailed information on seasonal reproduction patterns in mosquitofish in their native range and compare patterns between fish from the two lakes. Male mosquitofish were reproductively active from spring through fall. Spermatogenesis, which is regulated in part by 11-ketotestosterone, ceased in October, and fish stored spermatozoa through the winter for immediate fertilization of offspring in the spring. Compared with Lake Woodruff, fish from Lake Apopka tended to be larger and have longer gonopodia and greater gonado- and hepato-somatic indices (GSI and HSI). High GSI in Apopka fish correlated with greater spermatid production, but fewer mature spermatozoa and either the same or lower sperm counts and sperm viability. Taken together, these observations suggest that differentiation of spermatids to spermatozoa is disrupted in Apopka fish, leading to reductions in fertility in some months. Delivery of sperm to females could also be affected in Apopka fish, which exhibit lower prevalence of efferent duct tissue in the testes during the summer.  相似文献   

12.
The tench Tinca tinca is an interesting fish from the viewpoint of polyploidy and related atypical reproduction aspects. Triploid tench were produced artificially. Studies of spermiation as well as of sperm motility and structure were performed on several triploid and diploid males simultaneously with individual experimental crosses with diploid females to define their reproductive capacities. The testes of triploids visually looked less developed in the most of cases with lower sperm production (0.05 cm3 sperm per male), GSI and weight of testes compared to diploids (0.58 cm3 sperm per male). Analysis of variance showed significant influence of ploidy level on the percentage of motile spermatozoa. Triploidy did not change percentage of live spermatozoa and velocity of spermatozoa at the first time of sperm movement. The study of sperm structure by scanning electron microscopy revealed that most sperm cells were of normal structure with some anomalies. Sperm heads of triploid and diploid males were mostly round-shaped, 1.86±0.2 and 1.6±0.18 μm in diameter. The midpiece of triploid spermatozoa was slightly narrower than that of diploid ones with typical cylindrical shape. Flow cytometry revealed sperm cells of triploids to be largely aneuploid (1.47 n) with high mosaic DNA, oscillating from haploid DNA content (1.0 n) to diploid DNA content (1.9 n). Experimental crosses between triploid males and diploid females revealed that these males were capable to stimulate effective development with relatively high level of fertilization and hatching rates from 0 to 70%. In conclusion, triploidization does not seem to guarantee sterility of tench.  相似文献   

13.
The effects of dietary lipid from four experimental diets on the fatty acid (FA) composition and cholesterol (CHOL) content of spermatozoa and spermatozoal plasma membranes and their consequences for sperm viability after cryopreservation were evaluated in rainbow trout Oncorhynchus mykiss (Walbaum). The four sources of lipid were herring oil (adequate n‐3 FA), menhaden oil (high n‐3 FA), safflower oil (high n‐6 FA) or tallow (high saturated FA), and they comprised 12% of the total diet. Spermatozoa from fish fed the tallow diet had significantly (P < 0.05) higher CHOL levels than spermatozoa from the fish fed the other diets. The spermatozoal plasma membranes from fish fed the tallow diet had significantly (P < 0.05) higher CHOL and monounsaturated fatty acid levels than those from fish fed the menhaden or safflower oil diets, but were not different from membranes of fish fed the herring oil diet. Cryopreserved spermatozoa from fish fed the tallow or herring oil diets exhibited less membrane damage (P < 0.05) and produced a higher percentage (P < 0.05) of eyed embryos compared with spermatozoa from the menhaden or safflower oil‐fed fish. Therefore, it would appear that high levels of CHOL and monounsaturated fatty acids provided the spermatozoa with increased resistance to cryopreservation damage.  相似文献   

14.
Cryopreservation of sperm in marine fish   总被引：10，自引：0，他引：10  
Since the first work of Blaxter in 1953, fish sperm cryopreservation has been attempted on about 30 marine species. The present paper reviews the techniques used and the results published in these species. Particular attention is paid to the handling procedure of sperm before freezing, the problems of semen ageing and semen contamination with urine. The quality of frozen–thawed semen was evaluated using previously standardized biotests, such as a two‐step motility activation technique adapted for the different species and fertilization assays using a discriminating insemination technique. Most extenders used in marine fish are saline or sugar solutions. From the investigated cryoprotectants, dimethyl sulphoxide (DMSO) generally leads to the best results. Cooling rates range from 8 °C to 99 °C min?1; the thawing rate is generally high. Compared with freshwater species, a high percentage of spermatozoa survives cryopreservation. Therefore, and because of the simplicity of the techniques, the cryopreservation of marine fish sperm is suited for application in aquaculture.  相似文献   

15.
The protein composition of seminal fluid, blood serum, sperm plasma membrane and flagellum of rainbow trout were analysed by SDS-polyacrylamide gel electrophoresis. Immunological identity between proteins of the 2 fluids and sperm components was studied using crossed immunoelectrophoresis, rocket immunoelectrophoresis and immunoblotting. Results indicate that many seminal proteins are antigenically-related to serum proteins, proteins of sperm origin are present in seminal fluid in varying amounts, depending on the animals and sampling time, and several serum-like seminal proteins are bound to spermatozoa. Lipoproteins were isolated from seminal fluid (mean level: 33 μg/ml) and characterized. They were identified as being HDL-like lipoproteins. A possible physiological role is proposed for these seminal lipoproteins.  相似文献   

16.
Three groups of captive-reared striped bass Morone saxatilis ages 1, 3 and 12 yr, were examined for age-related changes of sperm characteristics including short-term storage. All groups had similar ranges of the following parameters (mean× SEM): expressible milt (5.6× 0.5 mI/kg body weight (BW) to 7.5× 2.1 mL/kg BW), percentage of motile sperm (55× 6% to 60× 2%), duration of sperm motility (69× 3 sec to 72× 5 sec) and percentage of viable sperm (91× 2% to 93× 2%). Compared to the 1 and 12-yr-old fish, the 3-yr-old fish produced the greatest number of spermatozoa (1,190× 370× 109 spermatozoa/kg), sperm concentration (120× 8 × 109 spermatozoa/mL) and spermatocrit (74× 4%). In addition, during short-term storage at 4 C, extender-preserved sperm samples of the 3-yr-old group showed a significantly higher ( P < 0.05) percentage of motile sperm and duration of sperm motility, compared to the other two groups. This suggests that short-term storage may be affected by the age of the male fish. Sperm longevity of the 3-yr-old group was successfully maintained for as long as 15 d, longer than that of the 1-yr-old group (9 d) and 12-yr-old group (7 d). Overall, the 3-yr-old fish appeared to have superior sperm quality than the 1 or 12-yr-old fish based on higher sperm production and increased sperm longevity.  相似文献   

17.
The Brazilian freshwater fish diversity is the richest in the world. Only 0.7% of all Brazilian species have had any aspect of their sperm biology addressed up to this date. The majority of the fish species described in this review migrate during the spawning season (a phenomenon known as piracema). Urbanization, pollution, hydroelectric dams and deforestation are some of the causes of stock depletion or even local extinction of some of these species. The knowledge concerning sperm quality and minimum sperm:egg ratio is important to maximize the use of males without reducing hatching rates. Furthermore, sperm cryopreservation and gene banking can guarantee the conservation of genetic diversity and development of adequate breeding programs of native fish species. In this review, we present and evaluate the existing information on Brazilian fish species that have been subject to sperm quality and cryopreservation studies. The following parameters were evaluated: volume of extractable sperm, sperm motility, sperm concentration, freezing media, freezing methods, and post-thaw sperm quality. Although the existing protocols yield relatively high post-thaw motility and fertilization rates, the use of cryopreserved sperm in routine hatchery production is still limited in Brazil.  相似文献   

18.
We conducted histological observation of male germ cells and reproductive organs of the starspotted smooth-hound Mustelus manazo in Tokyo Bay to reveal any abnormality in male reproductive traits, as part of a study to elucidate the factors causing recent fluctuation in abundance of the population. Spermatogenesis proceeded in spermatocysts from the germinal zone in the ventral part of the testis to the degenerative zone in the dorsal part, where the spermatozoa were conveyed into the ciliated lumina of the attached terminal branches of the intratesticular ducts. The intratesticular ducts were classified from their terminal ends into branch, stem, and collecting tubules. The ducts formed in the germinal zone and grew as the spermatocysts developed. An opening formed through the wall of each of the most mature spermatocysts into a branch tubule; bundles of spermatozoa were evacuated through this opening into the branch and then the stem tubule and subsequently into the collecting tubules in the rete testis and the efferent duct connected to the epididymis. Spermatocysts that were unable to emit sperm because of failure of adhesion to the branch tubules were disorganized in situ, as were their spermatozoa. The collapsed spermatocysts seem to be cleared by hemophagocytosis with lymphocytes and leukocytes, which may have been recruited from the epigonal organ. There were no specific abnormalities in the spermatogenesis or the morphological structure of testes, which suggested that an abnormality of male reproductive traits was not the major cause of the recent fluctuation in the population abundance of this species. Details of the intratesticular duct system for sperm emission to the epididymis are the first findings in elasmobranchs worldwide.  相似文献   

19.
This paper provides data concerning biochemical composition of milt of two sturgeon species, Siberian sturgeon bred in aquaculture facility of Inland Fisheries Institute in North Poland and sterlet (from two different populations from Danube and Odra). Milt plasma of Siberian sturgeon and sterlet, when compared to teleost fish, is characterized by much lower osmolality (up 70 to 96 mOsm kg−1) and protein concentration (0.24–0.58 g l−1). In spite of the presence of an acrosome and acrosin the antiproteinase activity of seminal plasma was low (12.79–25.40 U l−1). Activities of arylsulfatase and β-N-acetylglucosaminidase were found in spermatozoa. This agrees with the presence of an acrosome in sturgeons sperm. Similarly to mammals, these enzymes are also present in milt plasma. We determined a range of enzymatic activities from the minimal (seminal plasma) to the maximal damage (supernatants obtained after freezing-thawing without cryoprotectant). Activities of arylsulfatase, β-N-acetylglucosaminidase, lactic dehydrogenase and acid phosphatase were released from spermatozoa after freezing-thawing. For this reason they are good potential candidates as a markers of cryoinjury to sperm acrosome and midpiece. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

20.
Rainbow trout (Oncorhynchus mykiss) males were fed, during a 9‐month period, five experimental diets where fishmeal proteins were gradually replaced with cottonseed meal (CS) proteins (0, 25, 50, 75 and 100%; diets 1–5, respectively). This study was carried out to evaluate the action and tissue concentrations of gossypol. Growth performance of fish was not affected with the increasing levels of CS in the diets. Haemoglobin concentration and haematocrit significantly decreased in fish fed 100% CS compared with other dietary treatments. Gonadosomatic index, plasma sex steroids (testosterone, 11‐ketotestosterone and 17,20β‐dihydroxy‐4‐pregnen‐3‐one) and sperm characteristics (concentration, motility, protein concentration and lactate dehydrogenase activity) were not negatively affected by increasing levels of CS. For the first time, a comprehensive study of gossypol concentrations, total and (+) and (?)‐enantiomers, in several tissues (liver, blood plasma, spermatozoa and seminal plasma) was performed. The concentrations of both (+) and (?)‐enantiomers significantly increased with increasing levels of CS in the diet. The highest concentrations were found in the liver of fish fed diet 5 (185 ± 18 μg g?1). In blood plasma, the concentrations of total gossypol were 10 times lower than the one found in the liver, but 10 and 100 times higher than the concentrations in the spermatozoa and seminal plasma, respectively. In all tissues studied, the concentration of (+)‐enantiomer was higher than the (?)‐enantiomer. The ratio (?)‐enantiomer/total gossypol did not change significantly with the increasing levels of CS in the diet. The results of the present study indicate that CS can be used over a period of 9 months to replace fishmeal proteins completely without compromising growth and reproduction of rainbow trout males.  相似文献   

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