Cytogenetic and histological studies of the brook trout, Salvelinus fontinalis (Mitchill), and the Arctic char, S. alpinus (L.) hybrids

Although brook trout and the Arctic char hybrids are able to reproduce, individuals with decreased fertility or even fish that are unable to produce any gametes have been also described. Abnormal gonadal development and disturbances in the gamete production in the char hybrid offspring may be triggered by the odd chromosome number and disturbances in their pairing during meiosis. To verify this hypothesis, cytogenetic examination and the gonadal histology analysis of the brook trout x Arctic char hybrids were carried out. Diploid chromosome number in the studied char (F₁) hybrids varied from 82 to 84 (FN = 99–102). Among 28 hybrids, 12 males, three females, nine intersex individuals and two sterile specimens were described. In the case of two individuals, gonads were not found. Diploid chromosome numbers in the males and intersex individuals varied from 82 to 84. Chromosome numbers in the females were 82 and 83 chromosomes. Two sterile fish exhibited karyotypes composed of 82 and 84 chromosomes. Predominance of the ovarian component in the intersex gonads and gonadal sex ratio distortion towards the males suggested hybrid females had problems with gonadal differentiation. However, the lack of the clear relationship between chromosome number and gonadal development in the studied hybrids did not support our hypothesis that odd chromosome number may be responsible for such reproductive disturbances in the hybrid individuals. We have presumed that sterility and intersexual development of the gonads may be caused by interactions between brook trout and Arctic char genes on the sex chromosomes and autosomes rather than unpairing of the parental chromosomes.     

Sperm quality analysis of normal season (NG) and out-season by photoperiod manipulation (PG) of male rainbow trout broodstock (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

Sperm quality parameters in rainbow trout (Oncorhynchus mykiss) were investigated during normal season spawning (November–January) and out-season spawning (July–August) treated with artificial photoperiod manipulation. Normal spawning males (n?=?15) were kept in an open concrete pond under natural condition. Out-season spawning males (n?=?15) were treated with artificial LED light (50 lm/m²) in a closed concrete pond. In these two experimental groups, five fish were used in each of three spawning periods. The mean weight and body length of males (2?+ years, n?=?30) were 1213.43?±?39.43 g and 45.08?±?0.62 cm, respectively. Sperms were collected from July to August 2016 in the out-season spawning or photoperiod-manipulated group (PG) (water temperature 14.21?±?0.31 °C) and from December 2016 to January 2017, in the normal season spawning group (NG) (water temperature 8.81?±?1.03 °C). Volume of sperm, osmolality of seminal plasma, density of sperm, percentage of motile spermatozoa (MOT), curvilinear velocity (VCL), and duration of motility were measured for each male. Seminal plasma osmolality, density of sperm, and the motility of duration were 358.47?±?37.24 and 308.87?±?44.09, 4.37?±?2.10 and 9.8?±?1.56, and 8.8?±?2.42 and 24.6?±?6.76 in PG and NG, respectively. Fertilization rate was 37.79?±?9.37% and 94.51?±?1.33% in PG and NG, respectively. Sperm quality parameters showed significant differences in most of the cases (p?<?0.05) and fertilization rate at eyed egg stage (150–160 degree-days) was significantly higher in normal season spawning group than the photoperiod-manipulated group (p?<?0.05). Though the rate of fertilization was low in out-season, it was able to get enough gametes in summer using only artificial light having no changes in other parameters.     

The relationship between channel catfish female body weight and relative fecundity and fry production when induced to ovulate with carp pituitary extract and fertilized with blue catfish sperm

ABSTRACT

Little is known regarding the correlation between channel catfish female body weight and relative fecundity and fry production when the females are induced to ovulate with carp pituitary extract and fertilized with blue catfish sperm. A total of 145 female channel catfish were used in the early, middle, and late spawning season. Female body weight of channel catfish had negative correlations with relative fecundity (r = ?0.33 and ?0.25, P < .05; r = ?0.23, P > .05) and before the late spawning season with fry/kg (r = ?0.21, P > .05; r = ?0.30, P < .05). Eggs/kg female body weight and fry/kg female body weight followed a parallel pattern throughout the season, illustrating the logical relationship between relative fecundity and fry output. The size of females used is a consideration to maximize hybrid catfish embryo production during the early and middle spawning season but not late in the spawning season. However, these correlations are relatively small even though significant; thus a dramatic change in fry production would not be predicted. Since reproductive relationships can change during the spawning season, conclusions and recommendations could be flawed if data are not collected periodically during an entire spawning season.     

Sperm output in rainbow trout (Salmo gairdneri) — Effect of age,timing and frequency of stripping and presence of females

An experiment was conducted to investigate the sperm production of rainbow trout. Eighty-four first-spawners were allocated randomly to three groups to be stripped at weekly, 2-weekly or 4-weekly intervals, respectively. Half of each group were exposed to females. The trial extended over two successive spawning seasons.First-season spawners were modest sperm producers. Depending on the collection frequency they produced mean stripped semen volumes of 1.17 to 1.48 ml containing 7.7 × 10⁹ to 16.8 × 10⁹ sperm. The corresponding means for the second spawning season were 3.54 to 4.59 ml and 22.2 to 35.4 × 10⁹. The spawning season extended from October to April. Production and motility increased from the beginning to mid-season, followed by a gradual decline. The effect of the presence of females was not very evident. With increased frequency of stripping, more sperm/male were obtained. Total semen volume and number of sperm/spawner obtained during the first season with weekly, 2-weekly or 4-weekly collection amounted to 24.6 ml and 160.9 × 10⁹ sperm, 13.4 ml and 135.8 × 10⁹ sperm and 8.9 ml and 101.1 × 10⁹ sperm, respectively. The corresponding values for the second season were 88.9 ml and 546.1 × 10⁹ sperm, 50.3 ml and 377.5 × 10⁹ sperm and 32.1 ml and 247.9 × 10⁹ sperm.From a practical point of view, weekly collection is feasible in second-season spawners, but a maximum of one stripping every 2 weeks appears appropriate for first-season spawners.     

Isolation, cloning, sequencing of brain type aromatase and its expression in male and female Wrasse, Pseudolabrus sieboldi

Pseudolabrus sieboldi, wrasse being a diurnal spawner provides a good opportunity to study the endocrine mechanism of estrogen formation in brain and gonads. Moreover, an extremely large amount of E2 was produced in serum and testis of wrasse. It is assumed that the presence of E2 may play a major role in diurnal gametogenesis in male fish. In this study brain type aromatase have been isolated, cloned and sequenced from the brain of wrasse. Further, the expression pattern of brain type aromatase in gonads and adult tissue of male and female fish have been analyzed. In addition, the diurnal expression pattern of brain type aromatase in both male and female fish brain during spawning season have been analyzed. The P450arom (br) was isolated, cloned and sequenced from both male and female bamboleaf wrasse. The P450arom (br) gene (1877 sequenced nucleotide) contains an ORF of 1470 bp, a 5′-UTR of 18 bp and at least 407 bp in 3′-UTR. The amino acid sequence homology in the coding region of wrasse P450arom (br) is high compared to that of medaka, Oryzias latipes (80%), rainbow trout type 2, Oncorhynchu mykiss (78.2%), fugu, Takifugu ribripes (78%) rainbow trout type 1, (76%), goldfish, Carassius auratus (66.8%) and zebrafish, Danio rerio (66.2%). Expression study reveals that P450arom (br) mRNA were most abundant in brains of both male and female fish throughout the day during the spawning season. RT-PCR study revealed that P450arom (br) was expressed in skin, anal fin and tail fin of both male and female wrasse. P450arom (br) was not detected at any time of the spawning day in either ovary or testis of wrasse.     

Enhancement of reproductive performances of Gangetic leaffish, Nandus nandus through up regulation of serum Ca2+ concentration, improved morphological alteration of liver and ovary with dietary polyunsaturated fatty acids

Incorporation of polyunsaturated fatty acids (PUFAs) as biofunctional compounds with feed is an effective way for gonadal maturation without any hazardous effects on animal health, and thus it is possible to save the vulnerable species from the danger of extinction. In the present study sperm quality, level of Ca²⁺ concentration in serum, histological structure of the liver and developmental stages of ovary of an endangered fish species, Nandus nandus were investigated for the confirmation of the positive effects of PUFAs in reproduction and gonadal maturation. Fishes were collected from Brahmaputra River, Mymensingh, Bangladesh. Treated group was fed 1 % squid extracted phospholipid supplemented diet that was mixed with silver carp fish muscle where as controlled group was fed the same except phospholipid. For histology of liver and gonads, samples were dehydrated, cleaned and infiltrated, embedded in paraffin wax and sectioned. After that, the samples were stained with hematoxylin and eosin. The photomicrographs of the stained samples were taken by using light microscope. In comparison with the control group, treated group exhibited higher gonadal maturation which resulted in spontaneous spawning. Treated female demonstrated advanced gonadal developmental stages in comparison with the controlled female during different months. During spawning season, lipid granules and normal morphological alteration were observed in case of treated fish liver, whereas less lipid granules with more histological alteration of liver were observed in control group. Serum Ca²⁺ concentration in treated female was found significantly higher (P < 0.01) in contrast to the controlled female during the breeding season which was an indicator of the augment of estrogen secretion during ovarian maturation. Better sperm quality, early maturation of oocytes, less histological alteration of liver hepatocytes and spontaneous spawning performances of PUFA-treated fish were as a result of the efficiency of PUFAs in enhancing maturation. The experiment suggests that supplementation of dietary PUFAs improve the spawning performances of fish.     

Comparative ecologies of domestic and naturalised rainbow trout in northern Lake Huron

Northern Lake Huron (Ontario, Canada) supports the largest concentration of freshwater salmonid cage culture farms in Canada and receives inputs of domestic‐strain rainbow trout (Oncorhynchus mykiss) through escapement. We assessed the potential effects of these domestic fish on the naturalised rainbow trout of this ecosystem by comparing their respective ecologies. Mature adults were sampled from spawning tributaries, primarily in spring, and to a lesser extent in fall. Fish of domestic origin comprised ~80% of rainbow trout sampled from sites near cage culture facilities but <20% of rainbow trout sampled from more distant sites. Domestic adults in spawning condition (gametes free‐flowing) were present in the spawning tributaries in both spring and fall sampling seasons, whereas naturalised fish in spawning condition were only observed in the spring. Domestic adults were younger and smaller (in length), appeared to have shorter lifespan and had a higher male to female ratio compared with their naturalised counterparts. Growth rates (change in length with age) of naturalised and domestic females in the wild were similar, but domestic males grew more slowly than naturalised males. Domestic females also produced smaller eggs than naturalised females. Food web positions (inferred from δ¹³C and δ¹⁵N) of domestic and naturalised fish were very similar but varied more strongly with body size in the former. Domestic‐strain rainbow trout of cage culture origin can survive, grow and attempt to spawn in northern Lake Huron and have the potential to compete for food, mates and spawning habitat with naturalised rainbow trout.     

Cryopreservation and transplantation of sexually immature gonads of rainbow trout

The objective of this investigation was to cryopreserve sexually immature gonads of rainbow trout (RBT). Gonads from juvenile male or female RBT were cryopreserved and stored in liquid nitrogen. Individual testes were thawed and surgically transplanted to isogenic male trout; individual ovaries were autografted. Four weeks following transplantation, all male fish were injected with salmon pituitary extract for 8–10 weeks. The transplanted testis was present in 5 of 8 animals in replicate 1 and in 3 of 6 animals in replicate 2. Fertility of sperm obtained from individual testes in replicate 2 was evaluated by in vitro fertilization. Sperm from the intact control testes had a fertilization rate of 95±3% (mean ± SEM), and sperm from transplanted testes had a fertilization rate of 78±7%. Ovaries that were frozen in liquid nitrogen, thawed and autografted to an ectopic site were not present at five months following surgery.     

Reproduction related immunoglobulin changes in rainbow trout

Annual changes in plasma immunoglobulin (IgM) levels were investigated in three strains of rainbow trout Oncorhynchus mykiss which have different spawning periods, i.e., September–October, November–December, and January, reared under constant water temperature and natural day length. Plasma IgM levels decreased during the spawning season in all strains tested. The IgM changes became reversed in response to significant increases in plasma testosterone (T) and estradiol-17 in females and T and 11-ketotestosterone in males. Though the IgM decline showed a connection with suppressed immunocompetence, since many mature fish caught fungal diseases, no clear differences were observed in the plasma IgM levels between infected and noninfected fish during the spawning season. Incidentally, plasma IgM levels in infection prone fish were higher than in noninfection prone fish prior to the spawning season, whereas coincident differences in the plasma steroid levels were observed. Immature fish reared under lower water temperatures showed lower IgM levels. The effect of water temperature may have to be considered when analyzing the defense mechanism during the spawning season in rainbow trout.     

Variation in growth rate,liver weight and body composition at first sexual maturity in rainbow trout

Rainbow trout around 743 g were fed four different diets over an 8-month period. Maturing males grew faster in the period before spawning (Sept.–Dec.) than maturing females, and significantly faster than immature fish. Mature fish stagnated in growth while the immature fish surpassed the growth of mature fish during the spawning season. The condition factor was moderately influenced by the maturation process. In the period before spawning (Sept.–Jan.) maturing females had a significantly higher hepatosomatic index than males and immature fish. An increase in dry matter and fat and a decrease of protein were observed in the muscle of maturing males and immature fish towards the spawning season, while maturing females had a slight fall in dry matter and fat and a small increase in protein in the same period. Dry matter and fat in muscle of rainbow trout were positively correlated.     

虹鳟杂交胚胎(2n♀×3n♂)的发育状况及其DNA倍性分析

雄性三倍体虹鳟(Oncorhyncus mykiss)可以发育至生理成熟并表现出较强的雄性副性征,性腺发育基本正常；雌性三倍体虹鳟性腺几乎不发育,并且存在着类雄性化的发育趋势.本研究拟通过对虹鳟杂交胚胎(2n♀×3n♂)发育状况及其DNA倍性的分析,进而确认三倍体虹鳟是否具有与二倍体虹鳟类似的生育能力并分析其原因.选取经两个世代家系选育的虹鳟优良品系二倍体雌性虹鳟作为母本,以热休克方法人工诱导的优质雄性三倍体虹鳟为父本,常规人工采集精卵授精,获得三倍体雄性与二倍体雌性杂交胚胎,观察杂交胚胎的发育进程,通过流式细胞仪检测杂交胚胎的DNA含量,进而与二倍体雌雄交配获得的胚胎对照确定杂交胚胎的倍性.结果表明,杂交胚胎的受精率、发眼率和孵化率均显著地低于对照组,杂交胚胎多数在发眼期之前死亡,破膜的胚胎在仔鱼期上浮前全部死亡.杂交胚胎的DNA含量分布范围较大,大致可以分为两个水平,接近84％的胚胎的DNA含量介于二倍体与三倍体之间,约为对照组DNA含量的2.5倍；约有16％的胚胎的DNA含量在三倍体和四倍体之间.本研究认为,染色体非整倍体化可能会引起杂交胚胎细胞核质不相容,进而导致基因表达调控紊乱,从而使杂交胚胎不能正常发育,无法完成由母体卵黄供给的内源营养向外源营养的转换,成为导致杂交胚胎发育过程中大量死亡,以及杂交仔鱼在破膜之后开始进食之前全部死亡的一个重要原因.     

Early out-of-season induced spawning of channel catfish Ictalurus punctatus (Rafinesque) conditioned in heated earthen ponds

This study documents early out-of-season induced spawning of channel catfish Ictalurus punctatus. During the early spring (February–April) of 1999, 2000 and 2001, ponds containing (1) male and female channel catfish (mixed-sex ponds) or (2) male channel and blue catfish I. furcatus only, or female channel catfish only (single-sex ponds) were heated to 24–30°C to encourage gonadal maturation and spawning. Unheated ponds were stocked with males and females and were monitored during the duration of heating. When natural spawning occurred in the heated ponds, the fish were captured by seining and unspawned females were injected with 100 μg kg⁻¹ of synthetic leutenizing hormone-releasing hormone. Injected females were either paired with males or held in communal all-female groups, and monitored for ovulation. Eggs were collected and fertilized with sperm of channel catfish or blue catfish. Females paired with males were induced to spawn 44 days (mixed-sex ponds) and 50 days (single-sex ponds) before natural spawning occurred in unheated ponds. Spawning latency (the time between injection and ovulation) and the percentage of neurulated embryos from eggs fertilized using channel catfish sperm was not different between spawning before the natural season (P=0.68) and during the natural season in fish from mixed-sex ponds (P=0.57). Females held in all-female groups produced eggs 34 days before the onset of spawning in unheated ponds. Spawning latency was not different between spawns before and during the natural season (P=0.16), and the percentages of neurulated embryos from eggs fertilized with channel catfish sperm (P=0.76) or blue catfish sperm (P=0.77) before or during the natural season were not different. This study demonstrates the feasibility of conditioning of channel catfish females for early out-of-season induced spawning in the laboratory.     

Changes of sperm morphology,volume, density,and motility parameters in northern pike during the spawning period

Sexually mature males (BW?=?1600?±?150 g and TL?=?235?±?30 mm) of northern pike (Esox lucius L.) were randomly selected from a pond to record changes in their sperm quality parameters (spermatozoa morphology, sperm volume, density, and motility parameters) during the spawning season. The morphological and motility parameters changed significantly during the reproductive season with following trends. Only, head width was not changed during the spawning season. The longest spermatozoa and its flagellar length were found at the middle of spawning period (TL?=?38.24?±?0.37 μm and 35.14?±?0.26 μm) and shortest at the beginning of spawning period (TL?=?34.81?±?0.29 μm and 32.53?±?0.18 μm). Other morphological characters were always the lowest at the beginning of spawning period. Sperm volume was changed from 0.33?±?0.3 ml in February, 0.43?±?0.2 ml in March to 0.24?±?0.1 ml in April, and density from 16.2?±?0.2?×?109 spermatozoa ml^?1 in February, 19.4?±?0.2?×?109 spermatozoa ml^?1 in March to 4.8?±?0.2?×?109 spermatozoa ml^?1 in April. Same sperm velocity was observed in all spawning terms at 10 and 20 s after activation. Higher velocity was found at 30 and 40 s after activation in sperm collected at the middle and the end of spawning period. Significantly, higher percentage of motile sperm was observed at 20, 30, and 40 s after activation in sperm sampled at the end of spawning period. This study supports the hypothesis that longer spermatozoa swim faster.     

Quality of eggs and spermatozoa of rainbow trout fed an n-3 essential fatty acid-deficient diet and its effects on the lipid and fatty acid components of eggs, semen and livers

ABSTRACT: The effect of an n-3 essential fatty acid (EFA) -deficient diet on spawning and on the lipid and fatty acid contents of eggs, semen and livers of rainbow trout Oncorhynchus mykiss was investigated. Fish were split into two groups and fed either of two diets for a period of 4 months prior to the start of the spawning season. The control group was fed a commercial diet, containing n-3 EFA, whereas the experimental group was fed an n-3 EFA-deficient diet. Fish were 3 years old at the time of spawning. Eggs and semen were stripped off five females and five males from each diet group and cross-fertilized. Two of the five males fed the deficient diet showed a lower sperm motility, resulting in slightly lower mean hatching rates when crossed with eggs of either group. Higher lipid contents in the EFA-deficient diet were reflected in the egg and semen lipid contents, whereas the lipid contents of male livers of both diet groups were higher than those of female livers. In livers and eggs, the main polar lipid was phosphatidylcholine accompanied by phosphatidylethanolamine in sperm polar lipids. The non-polar lipids of eggs were mainly triacylglycerols but in livers and semen, free fatty acids and free sterols were more abundant. Essential fatty acids, namely n-3 highly unsaturated fatty acids and linolenic acid, were generally lower in eggs, semen and livers sampled from the EFA-deficient diet group. These results indicate that the fertilization, eyed and hatching rates obtained from crossings with males fed the EFA-deficient diet were slightly lower because of the dietary effect on sperm motility. Moreover, lipids of eggs, semen and livers of male and female rainbow trout were influenced greatly by their dietary availability.     

High‐throughput Cryopreservation of Sperm from Sex‐reversed Southern Flounder,Paralichthys lethostigma

The Southern flounder, Paralichthys lethostigma, is a valuable aquaculture fish with established markets in the USA. All‐female production in this species is an important technology for aquaculture because the females usually have body sizes twice those of males at the same age, and sex‐reversed males (genotypic XX neomales) are used for all‐female production by crossing with genetically normal females. However, sperm volume from the neomales is usually small (<0.5 mL) and limits their application for all‐female fish production. Cryopreservation of sperm from these sex‐reversed neomales will provide access on demand with increased efficiency to extend the application of neomales. The goal of this study was to develop a protocol for cryopreservation of sperm from the Southern flounder by using an automated high‐throughput processing system. The objectives were to: (1) determine the effect of osmolality on activation of sperm motility; (2) evaluate the effect of extender solutions on sperm motility capacity; (3) evaluate the acute toxicity of cryoprotectants (dimethyl sulfoxide [DMSO], propylene glycol, and polyethylene glycol) on sperm motility, and (4) estimate the effect of cooling rate on sperm cryopreservation and post‐thaw fertilization. Sperm motility was activated when osmolality was 400 mOsmol/kg or higher. Of the three extender buffers tested, HEPES4‐(2‐hydroxyethyl)‐1‐piperazineethanesulfonic acid (HEPES) at 300 mOsmol/kg resulted in better protection for sperm motility than did Hanks' balanced salt solution and Mounib solution at 300 mOsmol/kg during 7 d of refrigerated storage. After 30 min equilibration with the cryoprotectant of 15% DMSO, sperm motility was 24 ± 21% (fresh sperm motility without any cryoprotectants was 42%). After cooling at a rate of 20 C/min, post‐thaw sperm motility was 8 ± 5% and fertilization was 63 ± 40% evaluated at the 32–64 cell stage (5 × 10⁵ sperm per egg). Overall, a protocol was developed for sperm cryopreservation in the Southern flounder with high‐throughput processing, which provides a tool to preserve the valuable genetic resources from neomale flounders, and enables germplasm repository development for the Southern flounder.     

Diurnal rhythm of steroid biosynthesis in the testis of terminal phase male of protogynous wrasse, Pseudolabrus sieboldi, a daily spawner

The wrasse, Pseudolabrus sieboldi, is a diandric protogynous labrid fish. Spawning is performed by a terminal phase (TP) male and an initial phase (IP) female between 6:00 and 9:00 h daily during two-month-long spawning season. In the present study, to investigate the roles of steroid hormones in the diurnal spermatogenesis of the P. sieboldi TP male, all steroid hormones produced in the testis were identified and the synthetic pathways of these steroids were determined. Furthermore, the circulating levels of the major steroids produced were analyzed throughout a day at 3-hour intervals during spawning season. In the testis, 11-ketotestosterone (11-KT), estradiol-17β (E2), 17,20β-dihydoxy-4-pregnane-3-one (17,20β-P) and 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) were synthesized as the major metabolites. In vitro steroid biosynthesis experiments showed similar results to the circulation profiles of the major steroids. This study is the first to clarify the complete steroidogenic pathways in the gonads of a diandric protogynous species throughout its life, when combined with the results of the steroidogenesis in the ovarian follicles. This is also the first report of a clear diurnal rhythm of the steroid production corresponding to the spermatogenic process in the testis of a male teleost.     

Morphology,sex steroid level and gene expression analysis in gonadal sex reversal of triploid female (XXX) rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

In non-mammalian vertebrates, estrogens and expressions of cyp19a1 and foxl2 play critical roles in maintaining ovary differentiation and development, while dmrt1 and sox9 are male-specific genes in testicular differentiation and are highly conserved. In order to deeply understand the morphological change, sex steroids level and molecular mechanism of triploid female gonadal reversal in rainbow trout, we studied the ovary morphology, tendency of estradiol-17β (E2) and testosterone (T) levels and the relative expressions of dmrt1, cyp19a1, sox9 and foxl2 in juvenile and adult fish. Our results demonstrated that the development of triploid female gonads in rainbow trout went through arrested development, oocytes dedifferentiation, ovary reconstruction and sex reversal finally. During early gonadal development (154–334 days post-fertilization), the expressions of foxl2 and cyp19a1 increased linearly, while expressions of dmrt1 and sox9 were extremely suppressed, and E2 level was higher, while T level was lower. During the mid-to-late period of triploid female gonadal development (574–964 days post-fertilization), the expressions of dmrt1 and sox9 remained high and were very close to the quantity of diploid male genes, and T levels were even reaching diploid male plasma concentrations, while expressions of cyp19a1 and foxl2 were decreased, leading to decrease in E2 level. We realized that the development model of rainbow trout triploid female gonads was extremely rare, and the regulatory mechanism was very special. Genes involved in gonadal development and endogenous estrogens are pivotal factors in fish natural sex reversal.     

Gonadotropin releasing hormone‐analogue (GnRHa) treatment improves the milt production and sperm motility of endangered Caspian brown trout,Salmo trutta caspius,over the course of a spawning season

To determine the effect of gonadotropin releasing hormone‐analogue (GnRHa) treatment on the milt quality of endangered Caspian brown trout, Salmo trutta caspius, the sperm motility (percentage and duration of motility), sperm production (sperm density, spermatocrit and milt volume) and milt pH were measured for GnRHa‐treated (the treatment group) and untreated groups (the control group) during the spawning season. For untreated brooders, the values of the motility per cent, sperm density and spermatocrit decreased continuously during the spawning season while the milt volume, duration of motility and milt pH showed only a significant decrease at the end of the season. For GnRHa‐treated males, these parameters increased 14 days after GnRHa treatment (first milt collection) and then decreased continuously towards the end of the season. In addition, the values of milt and sperm density yielded per treated male were higher than that in the untreated group, although these were not statistically different. In any case, the total sum of yielded milt from the treatment group over the spawning season was higher than that in the untreated group. In this experiment, significant positive correlations were found between milt parameters as follows: sperm motility vs. milt pH; sperm density vs. spermatocrit; milt volume vs. spermatocrit; and milt volume vs. sperm density. The results show that the treatment of Caspian brown trout by GnRHa can improve the milt quality in terms of sperm motility and sperm production during a spawning season.     

Changes in Sperm Production,Sperm Motility,and Composition of Seminal Fluid in Caspian Brown Trout,Salmo trutta caspius,Over the Course of a Spawning Season

This study was carried out to evaluate milt quality in male Caspian brown trout (Salmo trutta caspius) over the course of the winter spawning season. Milt samples were collected biweekly during December and January. Chemical composition of seminal fluid, sperm production (milt volume, sperm density, spermatocrit,) and sperm motility characteristics (percentage and duration of motility) were measured. Milt volume, sperm density, osmolality, seminal minerals (Ca²⁺, Mg²⁺, K⁺, Na⁺, Cl^?), and total protein gradually decreased over the spawning season. Glucose and triglyceride content of milt did not show significant changes over the spawning season. Milt pH and the percentage and duration of motility were comparatively stable, declining only at the end of the season. Significant positive correlations were found between sperm density and seminal minerals, total protein and spermatocrit; percentage of motile spermatozoa and seminal minerals, total protein; and duration of motility and K⁺, Cl^?, total protein, and pH. Results show that season has a significant influence on milt quality in male Caspian brown trout, with the best milt being available at the beginning of spawning season.