Seasonal reproductive cycle of Waigieu seaperch (Psammoperca waigiensis)

Predictive and reliable parameters of reproductive status are integral aspects of sustainable fisheries and aquaculture management. These parameters are also important for an accurate evaluation of the effects of different treatments on sexual maturation in fish farming. In the present study, we have characterized the seasonal reproduction profile and described changes in sex steroids in relation to gonadal maturation and development in female and male Waigieu seaperch (Psammoperca waigiensis). The experimental period covered a full calendar year (January–December). In males and females, we observed that plasma sex steroid hormones [oestradiol‐17β (E2), testosterone (T), 11‐ketotestosterone (11‐KT) and progesterone (P)] levels showed monthly fluctuations during the spawning period. Particularly, plasma steroid hormone levels were positively associated with gonadosomatic index values. In addition, high levels of plasma steroid coincided with recruitment of oocytes into yolk accumulation in females. The main spawning period occurred between April and October in females, and between March and November in males. The non‐aromatizable androgen, 11‐KT is generally believed to be the active male‐specific androgen in teleosts, and is associated with the process of spermiation, development of secondary sexual characteristics and regulation of male reproductive behaviour in most teleost species. In this study, we found relatively high amounts of 11‐KT in females between May and December, suggesting an integral role in the maturation process, also for the females. A rapid peak in plasma P level was observed in November and suggests significant roles during post‐spawning and/or resting periods in both female and male fish. Furthermore, all oocyte developmental stages were present within the same sampling month and also within the spawning period, demonstrating the gamete group asynchronous developmental strategy. Overall, Waigieu seaperch showed strong seasonality in reproductive development with corresponding sex steroid patterns. The data presented in this study may contribute to the understanding of the reproductive endocrinology of a tropical marine finfish with increasing industrial prospects and sustainable aquaculture of this species in a developing country, such as Vietnam.     

Sperm Motilities in Waigieu Seaperch,Psammoperca waigiensis: Effects of Various Dilutions,pH, Temperature,Osmolality, and Cations

The objectives of this study were to evaluate the effects of varying dilutions, pH, temperature, osmolality, and cations on sperm motility parameters in waigieu seaperch, Psammoperca waigiensis. The maximum velocity of average in path (VAP), percentage of motile cells (MOT), and duration of sperm motility (DSM) were observed when semen was diluted in artificial seawater (ASW) at a ratio of 1:100 (144.9 ± 0.6 µm/sec, 95.6 ± 0.4%, and 230.3 ± 2.3 sec, respectively), at 30 C (142.0 ± 0.6 µm/sec, 93.6 ± 0.4%, and 238.3 ± 0.9 sec, respectively), and pH 8 (144.8 ± 0.6 µm/sec, 93.3 ± 0.4%, and 234.0 ± 1.5 sec, respectively). Maximum VAP, MOT, and DSM were obtained in each solution containing 0.6 M NaCl (143.8 ± 1.0 µm/sec, 91.3 ± 2.0%, and 230.6 ± 4.2 sec, respectively), 0.6 M KCl (135.1 ± 3.1 µm/sec, 91.1 ± 3.1%, and 230.3 ± 3.7sec, respectively), 0.2M CaCl₂ (105.3 ± 4.7μm/sec, 47.9 ± 2.7%, and 120.7 ± 1.3 sec, respectively), 0.2 M MgCl₂ (107.3 ± 3.0 m/s, 42.1 ± 3.3%, and 120.3 ± 4.8 sec, respectively), and osmolality of 400 mOsm/kg (145.1 ± 2.5 µm/sec, 93.0 ± 2.1%, and 346.5 ± 4.4s, respectively). We used these mediums as artificial insemination media for fertilizing matured eggs. The results showed that the fertilization and hatching rates in 0.6 M NaCl (75.3 ± 0.6% and 57.0 ± 2.4%, respectively), ASW (70.8 ± 1.2% and 51.2 ± 1.8%, respectively), or 400 mOsm/kg (72.9 ± 1.8% and 55.3 ± 1.6%, respectively) were higher than that in seawater (63.9 ± 1.2% and 39.2 ± 3.9%, respectively). In conclusion, using 0.6 M NaCl, ASW, or 400 mOsm/kg as an artificial insemination medium is effective for fertilizing of waigieu seaperch.     

Seasonal Changes in the Milt Quality of Waigieu Seaperch,Psammoperca waigiensis: Implications for Artificial Propagation

Milt quality of Waigieu seaperch was sampled at the beginning, the middle, and the end of the spawning season to assess if the sampling season has an effect on milt quality parameters. The milt volume and total sperm production were higher at the middle of the spawning season while the sperm concentration in milt was significantly higher at the beginning and the end of the spawning season. Sperm morphology and the major parameters of seminal plasma (pH, total protein, Mg²⁺, or Ca²⁺ concentrations) did not differ throughout the spawning season. The concentrations of Na⁺ and the osmolality increased throughout the spawning season. The percentage of motile cells did not differ during the spawning season while the duration of sperm motility (swimming duration) and velocity (swimming speed) were significantly different during the spawning season. The fertilization rate (75.7 ± 6.8%) and hatching rate (56.5 ± 3.1%) at the middle of the spawning season were higher than at the beginning and end periods. Results indicated that milt quality parameters and fertility success in the middle of the spawning season were higher compared to earlier and later sampling dates. Thus, we recommend collecting milt during this time for maximal fertilization and hatching success.     

Effects of steroid hormones on in vitro oocyte maturation in white sturgeon (Acipenser transmontanus)

The in vitro effects of several steroids on the maturation of intact white sturgeon (Acipenser transmontanus) ovarian follicles were investigated. At the highest concentration (1024 ng ml^–1 for the C21 steroids and 1139 ng ml^–1 for the C19 steroids), all of the C21 steroids tested, progesterone (P4), 17-hydroxyprogesterone (17OHP), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,(20,21-trihydroxy-4-pregnen-3-one 20-S), 11-deoxycortisol (S) and cortisol (F), as well as testosterone (T) induced germinal vesicle breakdown (GVBD) at 14 and 22 h. At 6 h, only P4 and 17,20-P induced maturation at the highest concentration (1024 ng ml^–1). At 14 and 22 h, 11-deoxycortisol was the most potent steroid inducer of GVBD followed by P4, 17OHP, 17,20-P, and 20-S. The steroid 11-hydroxytestosterone (11OHT) was completely ineffective at all concentrations and exposure times. The C21 steroids induced oocyte maturation at concentrations ranging from 4 to 1024 ng ml^–1, whereas T induced GVBD at 225 to 1139 ng ml^–1. Calculation of the mean effective concentration that induced 50% GVBD (EC50) from the 22 h incubations revealed the following order of potencies: S > P4 > 17OHP > 17,20-P > 20-S >> F > T. These bioassay results, together with previous findings on the endogenous production of steroids by ovarian follicles from gonadotropin-primed females, indicate that more than one steroid has a biological role in the resumption of meiosis in sturgeon oocytes and provides empirical evidence for P4, 17OHP, S, 20-S, and 17,20-P as maturation-inducing steroids in white sturgeon.     

Effects of the aromatase inhibitor Fadrozole on plasma sex steroid secretion and oocyte maturation in female coho salmon (Oncorhynchus kisutch) during vitellogenesis

17-estradiol, 17-20-dihydroxy-4-pregnen-3-one (17-20-P), and testosterone levels were measured in plasma samples obtained from vitellogenic coho salmon (Oncorhynchus kisutch) before and 32 days after injection of the aromatase inhibitor Fadrozole (AI). Plasma 17-estradiol levels decreased significantly 6 h after injection in all AI treated fish. The higher the dose the longer the maintenance of low plasma 17-estradiol levels. Inversely, plasma 17-20-P increased significantly 6 h after injection in all AI treated fish, and the higher the dose the longer the maintenance of high plasma 17-20-P levels. At 48 h after injection plasma testosterone levels were significantly higher in the AI treated groups. The oocyte maturation index showed that multiple injections with AI retarded oocyte development. Besides, oocyte diameter and GSI were lower in the same group, which presented high incidence of atresia of vitellogenic oocytes. The ovarian follicles and brain of the fish which received multiple injections secreted less 17-estradiol, in vitro. These findings suggest that aromatase inhibitors such as Fadrozole may have a potential as a tool to regulate sexual development in salmon.     

Effects of insulin-like growth factor-I on final oocyte maturation and steroid production in Fundulus heteroclitus

Recent evidence has indicated the presence of IGF-I and IGF-I receptors in mammalian and teleost ovarian follicles. Since growth hormone (GH), which can be secreted from the pituitary concomitant with a gonadotropin as a response to gonadotropin-releasing hormone, generally acts to release IGF-I from tissues including the ovary, the effect of IGF-I itself on ovarian steroidogenesis and oocyte maturation was investigated in the model teleost, Fundulus heteroclitus. IGF-I was found to be without effect on ovarian follicle steroidogenesis, but initiated oocyte maturation in a dose-dependent manner even more rapidly and effectively than 17,20-dihydroxy-4-pregnene-3-one (DHP), the naturally occurring maturation-inducing steroid. IGF-II also induced oocyte maturation in a dose-dependent manner. IGF-I induction of oocyte maturation occurred in the absence of DHP production by the granulosa cells (which is normally stimulated by gonadotropin), and could be inhibited by cycloheximide but not actinomycin D, thus implicating the role of protein synthesis. These results suggest that GH-stimulated release of ovarian IGF-I may have an even more direct role than DHP on the reinitiation of oocyte maturation.     

Influence of pejerrey Odontesthes bonariensis (Valenciennes, 1835) broodstock age on gamete quality,reproductive performance and plasma sex steroid levels during the spawning season

This study evaluated the effect of pejerrey Odontesthes bonariensis (Valenciennes, 1835) broodstock age on gamete and spawning quality and its relationship with sex steroid plasma levels. Sperm was analysed using a computer‐assisted sperm analysis (CASA) developed for pejerrey. Semen samples were taken from all fish from mid age groups (5‐ and 7‐year fish), but it was not possible to obtain stripped semen samples from all fish of younger (3‐year) or older (10‐year) groups. The highest relative sperm weight and sperm concentration were recorded in 5‐ and 7‐year groups respectively, while viability was not different between age groups. It was not possible to identify an effect of age on sperm quality by CASA. Non‐viable spawned eggs were obtained in the 3‐year group and, the relative fecundity and the relative number of fertilized eggs decreased with age in other groups. Fertilization and hatching rates showed the highest values in the 10‐year group. However, the estimated number of hatched larvae was similar in 5‐, 7‐ and 10‐year fish. A clear effect of age on 11 ketotestosterone (11‐KT) and estradiol (E₂) levels was identified, with the highest values in the 5‐ and 7‐year groups. These results might be related to the onset of puberty in the 3‐year group and ageing in the 10‐year group. Considering that the estimated number of larvae obtained was similar among age groups, the results of this study suggest that broodstock maintenance cost could be reduced by using mid‐age fish rather than older fish in pejerrey hatcheries.     

In vitro steroid metabolism during final oocyte maturation in white croaker Micropogonias furnieri (Sciaenidae)

The main steroids produced by three stages ovarian fragments (post-vitellogenic PV, oocytes in GV migration phase Mtg and mature oocytes M) of white croaker were compared. In Mtg and M stages 17,20β-dihydroxy-4-pregnen-3-one (17,20βP), 17,21-dihydroxy-4-pregnene-3,20-dione (17,21P) and 17,20β,21-trihydroxy-4-pregnen-3-one (20β-S) were synthesised. The specific synthesis of 20βS in Mtg oocytes suggests a MIS role for this steroid.     

饲料中花生四烯酸对发育前期大菱鲆亲鱼性类固醇激素合成的影响

为研究饲料中不同水平的花生四烯酸(arachidonic acid,ARA)对发育前期的大菱鲆亲鱼性类固醇激素合成量及合成过程的影响,配制3种等氮等脂(脂肪含量13%)的实验饲料,分别含有不同梯度水平的花生四烯酸:0.72%(不添加花生四烯酸精制油的对照组,C),5.63%(添加低水平ARA的处理组,ARA-L)及15.03%(添加高水平ARA的处理组,ARA-H)(各数值均为占总脂肪酸的比例)。每组饲料投喂3个实验桶,每桶放25尾3龄大菱鲆亲鱼(雌雄比例约为1:1)。养殖实验在室内流水系统内进行,每天饱食投喂2次,养殖周期为5个月。养殖结束后,分别取性腺发育前期的雌雄鱼测血清雌二醇和睾酮的含量并检测性腺中性类固醇激素合成相关蛋白质的基因表达量。结果显示:与对照组相比,饲料中高水平的花生四烯酸显著降低了雌鱼血清中雌二醇的含量,而雄鱼血清中睾酮的含量在低水平花生四烯酸处理组显著降低。在卵巢中,饲料中高水平的花生四烯酸显著降低了促卵泡激素受体mRNA的表达量,但是饲料中低水平的花生四烯酸显著提高了17α羟化酶的mRNA表达量。在精巢中,饲料中低水平的花生四烯酸显著降低了固醇合成急性调节蛋白以及17α羟化酶的mRNA表达量,但显著升高了芳香化酶的mRNA表达量。饲料中花生四烯酸提高了性腺、肝脏和肌肉组织中花生四烯酸的含量,但降低了二十碳五烯酸(EPA)的含量,卵巢中的花生四烯酸累积量高于精巢。综上所述,饲料中添加一定量的花生四烯酸抑制了发育前期大菱鲆亲鱼雌二醇和睾酮的合成,在卵巢中,这种抑制作用可能是通过抑制促卵泡激素受体的表达来实现,而在精巢中,可能是通过抑制固醇合成急性调节蛋白以及17α羟化酶来实现。     

Spawning induction and hormonal levels during final oocyte maturation in the silver perch (Bidyanus bidyanus)

The silver perch (Bidyanus bidyanus Mitchell) (Teraponidae), is a native Australian freshwater fish that, due to its high potential for aquaculture, was introduced to Israeli fish farming. The objective of this study was to find an optimal method for inducing spawning of silver perch. The agents tested for this purpose were: human chorionic gonadotropin (hCG; 150 or 200 IU/kg BW); salmon gonadotropin releasing hormone analogue (sGnRHa at 10, 20, 30, or 40 μg/kg BW); mammalian GnRH analogue (mGnRHa at 30 μg/kg) and the combination of sGnRHa at 20 μg/kg and domperidone at 5 mg/kg BW. Based on spawning success and relative fecundity, sGnRHa at the dose of 30 μg/kg was found to be more efficient than hCG, mGnRHa or sGnRHa with domperidone. Since domperidone does not improve the GnRHa effect on final oocyte maturation (FOM) and spawning, it is suggested that the dopaminergic inhibition during the stages of FOM in the silver perch is weak. Therefore, the use of GnRHa alone is sufficient to induce spawning in this fish. Immunoreactive gonadotropin (IR-GtH) and estradiol levels increased after a single injection of sGnRHa, and peaked after 24 h. Plasma levels of 17α,20β-dihydroxy-4-pregnen-3-one (17,20-P) also increased significantly 24 h after the injection of mGnRHa, 12 h before spawning, suggesting that 17,20-P is the maturation-inducing steroid in silver perch. In order to reveal whether the heterologous gonadotropin may elicit an immunological reaction, silver perch was subjected to prolonged treatment with hCG. This treatment resulted in no detectable titer of antibodies against the mammalian gonadotropin. In conclusion, although hCG has no deleterious effects in this fish, and is the more commonly used for spawning induction, sGnRHa at 30 μg/kg is the recommended treatment for spawning induction of female silver perch under the conditions prevailing in Israeli aquaculture.     

In vitro oocyte maturation and blood‐circulating steroid hormones in Sterlet (Acipenser ruthenus) and Siberian sturgeon (Acipenser baeri)

The oocyte of Sterlet (Acipenser ruthenus) and Siberian sturgeon (Acipenser baeri) was evaluated after incubation with 17α, 20β‐Dihydroxyprogesterone (DHP) in artificial media. Concentration of testosterone (T), progesterone (P4) and 17‐β estradiol (E2) in blood and their relation to fertilization rate were measured during experimental period. The oocytes at the same developmental stages were incubated in SIS (based on sturgeon blood ionic composition), RM2 (Ringer solution modified for sturgeons) and L‐15 (Leibovitz medium) artificial mediums in the presence of 1 μg/ml of 17α, 20β‐Dihydroxyprogesterone for 12, 18 and 24 hr. The result of this study demonstrated that fertilization rate decreased with the increasing duration of incubation in all mediums. In the Siberian and Sterlet sturgeon, the highest fertilization rate was observed in the oocytes incubated in RM2 (0.81 ± 0.4) and SIS (0.44 ± 0.8) mediums for 12 hr respectively. In the Siberian sturgeon, fertilization rate decreased significantly (p < .05) with incubation time in all three mediums. In Starlet, fertilization rate decreased significantly (p < .05) with time and reached a minimum after 24 hr (0.16 ± 0.1) in L‐15 medium. No significant (p > .05) differences were observed between blood plasma hormones and fertilization rate. The results of this study indicated that 17α, 20β‐Dihydroxyprogesterone is essential for induction of in vitro oocyte maturation in these species.     

Inducing oocyte maturation, ovulation and spawning in the African catfish, Heterobranchus longifilis Valenciennes (Pisces: Clariidae), using frog pituitary extract

Abstract. Oocyte maturation, ovulation and spawning were successfully achieved in the African catfish, Heterobranchus longifilis Valenciennes, using frog pituitary extract. During artificial spawning, oocyte maturation and ovulation were induced at a dosage of 7mg per kg of individual fish weight. Latency period varied from 7 h at 29°C to 11 h at 25°C. Dead eggs became opaque between 8 and 10h after fertilization while embryogenesis was found to be temperature dependent. Mean hatching and larval deformity rates were 63·08%± 7·56 and 8·08%± 1·93 respectively. During induced natural spawning, the extract was applied at a dosage of 35 mg per kg of female fish weight. Spawning occurred between 10 and 11 h after hypophysation. Mean hatching and larval deformity rates were 41·38%± 8·48 and 10·00%± 2·00 respectively.     

Effects of two sex steroid hormones on early oogenesis in Japanese huchen (Hucho perryi)

The present study was carried out to investigate whether several sex steroid hormones are able to induce oogonial proliferation or initiation of meiotic division I in vitro. The results showed that 17β-estradiol (E2) and 17α, 20β-dihydroxy-4-pregnen-3-one (17α, 20β-DHP) were able to induce the DNA synthesis of ovarian germ cells. The percentage of oocytes in 17α, 20β-DHP treated fragments was significantly higher than that in the other treatments. From these results, we conclude that endogenous E2 and 17α, 20β-DHP synthesized in ovary play significant roles in the proliferation of oogonia and the initiation of meiotic division I in early oogenesis, respectively.     

Influence of the spawning environment on final maturation and spawning in Japanese dace, Tribolodon hakonensis

Results of the present study demonstrated that a specific combination of spawning substrate and water current was indispensable for the spawning of mature Japanese dace, Triborodon hakonensis. It became evident that the environmental factors acted as a trigger for endocrine changes leading to final maturation in this species.     

Effects of dietary arachidonic acid on final maturation,spawning and composition of gonad of black sea urchin Diadema setosum (Leske, 1778)

This study investigated the effect of dietary arachidonic acid (ARA, 20:4n‐6) on final maturation, spawning performance and composition of the gonad of black sea urchin (Diadema setosum). Four different dietary ARA levels of 0.3 g/kg (Control diet), 1.4 g/kg (ARA 1.4), 2.5 g/kg (ARA 2.5) and 3.7 g/kg (ARA 3.7) were prepared and tested in this feeding trial for 90 days. The result showed that growth in terms of weight was not relatively affected by dietary ARA. Gonadosomatic Index (GSI) of animals were gradually increased in all the diets throughout the trial. The highest GSI level (25.7 ± 3.16%) was observed in animals fed with the diet ARA 1.4. ARA content in the gonad proportionally increased with dietary ARA levels. The highest larval survival rate (85.5 ± 5.67%) was found in diet ARA 2.5. The results of this study indicate that ARA plays an important role in the reproduction functions of black sea urchin and supplementing an appropriate level of ARA in the broodstock diet could induce the final maturation and improve reproduction of black sea urchin, but a higher level of ARA (3.7 g/kg) seems to have a negative effect on both growth rate and GSI.     

饲料脂肪源搭配对施氏鲟雌鱼繁殖性能和血清性类固醇激素的影响

以鱼油和玉米油作为脂肪源, 配制4种等氮饲料(蛋白含量38%, 脂肪含量12%~13%), 使脂肪酸中高度不饱和脂肪酸(n-3HUFA)的比例分别达3.1%、2.5%、1.9%、1.3%(分别简称SL1、SL2、SL3和SL4), 以其饲喂处于性腺发育Ⅲ期的施氏鲟(Acipenser schrenckii)亲鱼5个月, 研究不饱和脂肪酸对其繁殖性能及性类固醇激素水平变化的影响。结果表明, 经过 5个月养殖, 施氏鲟亲鱼平均体质量增加14.5%, 经过低温刺激及升温处理至产前体质量下降6.8%, 各组之间差异不显著(P>0.05); 饲料中n-3HUFA和n-3PUFA水平对繁殖性能有不同程度的影响, SL1组和SL2组亲鱼的成熟度和仔鱼成活率高于SL3和SL4组, 而仔鱼畸形率显著低于SL3和SL4组(P<0.05); SL1组极化卵卵径显著高于其他组(P<0.05), 平均产卵量高于其他组, 但各组之间差异不显著(P>0.05); 各组之间的受精率、孵化率差异不显著(P<0.05); 各饲料组间血清E₂和血清T含量差异不显著(P>0.05)。本研究表明, 使用混合鱼油作为饲料脂肪源, 效果好于单一使用鱼油或玉米油, 同时鱼油的需求比例大于玉米油, 建议n-3 HUFA的适宜添加量在2.5%左右, n-3 PUFA最适需要量需要进一步研究。

     

Effects of carotenoids and retinol in oocyte maturation of crayfish Cherax quadrucarinatus

Bioactive additives in food to improve growth, development and reproductive success of crustaceans have received little attention. In the present study, the effects of carotenoid and vitamin A injections were characterized and correlated to the ontogenic development of oocytes in female Cherax quadricarinatus reproducers. During an acclimation period, 250 C. quadricarinatus females were fed a basal diet devoid of carotenoids and retinoids. Four treatments were administered by injection: retinol palmitate (RP), β‐carotene, astaxanthin and vegetable oil as the control treatment. Before the experiment, total gonadic mass of organisms was obtained to determine the gonadosomatic index. Histological techniques were used to evaluate the effects of the different treatments during the primary vitellogenic phase. Retinol palmitate had the greatest inductive effect on the primary vitellogenic phase and on indicators of ontogenic oocyte development. The bioactive and molecular foundations of the action of carotenoids and retinoids that actively participate during the process of nuclear signalling and differentiation of the oocyte are discussed, considering these compounds as essential nutritional additives during gonadic maturation.     

南美白对虾的性腺发育、交配、交卵和受精

以池养南美白对虾（Penaeus vannamei）亲虾进行人工促熟，并使之交尾、产卵、受精。实验在上海金山区抗州湾河口低盐地区一套自行设计的水循环系统中进行，育苗用水通过兑配使盐度为27-28。育苗期间基本不换水和移动亲虾。亲虾205尾，雌雄比约1：1。雌虾切除眼柄后3-4d，性腺即开始发育，6-7d产卵，但未受精。手术后约30d，亲虾出现追尾现象、随之发生交配并出现受精卵。实验进行了约60d，共收集虾卵456.7万粒，其中受精卵出现后所产卵为137万粒，通过设置在水循环系统内的集卵网袋，可收集到95％以上的卵，共获受精卵61万粒，平均日受精率为44.5％，无节幼体25.1万尾，受精卵的平均孵化率为41.2％，最高达87.5％。文中还重点讨论了雌雄亲虾性腺发育的特点，产卵交配规律，以及亲虾规格大小、饵料种类、水质状况对南美白对虾交配和受精的影响。