Effect of exogenous hormones on ovulation and gonadal steroid plasma levels in starry flounder, <Emphasis Type="Italic">Platichthys stellatus</Emphasis>

The present study was designed to examine the potential for inducing ovulation in starry flounder (Platichthys stellatus) using gonadotropin-releasing hormone analog (GnRHa) and human chorionic gonadotropin (hCG) to assess whether starry flounder are differentially responsive to GnRHa and hCG. Female starry flounder were injected or implanted with different doses of hCG or GnRHa pellets to examine their ovulation-inducing potential and effects on plasma levels of testosterone (T), 17β-estradiol (E2), and 17,20β-dihydroxy-4-pregnen-3-one (17,20βP). Blood samples were collected for up to 10 or 25 days post-injection or post-implantation in two separate experiments designed to mimic the early and middle stages of spawning, respectively. Fish treated with the GnRHa pellets (100 µg) showed a significant increase in the total number of stripped eggs relative to the controls. GnRHa administration had no effect on the floating rate or fertilization rate of ovulated eggs in the both experiments, whereas hCG injection affected both of these rates. Plasma T levels were not significantly different between the exogenous hormone-treated and control fish. In contrast, the plasma E2 level was elevated in those fish treated with GnRHa, regardless of injection or implantation, and was accompanied by increased numbers of stripped eggs in both experiments. Treatment with GnRHa resulted in higher 17,20βP levels compared to the controls, and there was a positive relationship between elevated plasma 17,20βP and an increase in ovulated eggs in response to GnRHa treatment. The implantation of starry flounder with GnRHa-containing pellets was effective at inducing sustained ovulation compared to hCG treatment.     

Induced Ovulation of Southern Flounder Paralichthys lethostigma Using Gonadotropin Releasing Hormone Analogue Implants

Implanted pellets that provide a sustained release of [D-Ala⁶ Des-Gly¹⁰] LHRH-ethylamide (GnRHa) were used to induce maturation and ovulation of Southern flounder Paralichthys lethostigma . Of the 12 females whose ovaries contained follicles with a maximum diameter ≥500 μm, 11 ovulated for the first time within 90 h of hormone implantation. Only 1 fish with a maximum follicle diameter less than 500 μm ovulated within 2 wk after implantation. Ovulated eggs were manually stripped from the females and mixed with sperm from several males. Most females were spawned 1 to 3 times on consecutive days with variable fertility. One female was spawned 11 times producing 668,000 eggs. Fertility was evaluated by examining the incubated eggs for early stages of embryonic cleavage. The percentage of fertile eggs in subsamples of incubated eggs ranged from 7–95%. The results indicate that GnRHa implants can be used to induce repeated ovulation in this species. The variability in fertility is discussed in relation to egg quality.     

Reproduction of striped bass, Morone saxatilis (Walbaum), broodstock: monitoring maturation and hormonal induction of spawning

Abstract Levels of gonadal steroid hormones were quantified in an adult striped bass, Morone saxatilis (Walbaum), broodstock during their gametogenic cycle. Blood plasma concentrations of Estradiol (E₂) and testosterone in females, or 11-ketotestosterone(11-KT) and testosterone (T) in males, were used as indicators of maturation. In both sexes, hormone levels were low in summer but increased significantly by late October to intermediate levels which were then maintained until late January. They then increased again rapidly to maximum pre-spawning values attained in late February or March, and subsequently decreased during the spawning period (April and May) with an increased incidence of spent fish with low hormone levels. The changes in blood hormone concentrations coincided with annual changes in photoperiod and water temperature that may be useful landmarks for maturation in captive broodstock. Mature females were implanted with pellets containing a dose of approximately 20 μg/kg body weight of [D-Ala⁶-Pro⁹-Net]-LHRH (GnRHa) in a matrix of cholesterol (CH) and cellulose. In April, they had not yet begun final oocyte maturation (FOM) and were too immature for conventional induction of spawning by injection with human chorionic gonadotropin (hCG). In early April, females given two 95% CH (slow hormone-release) GnRHa pellets (95/95) or females given one 80% CH (fast hormone-release) GnRHa pellet and one 95% CH GnRHa pellet (80/95) spawned within 13 days treatment (n= 4) with good egg fertility (76 ± 7% of total) and hatch rates (62 ± 15% of fertile). Females given dual fast-release GnRHa pellets (80/80) or control (Sham) pellets did not spawn or show evidence of increased oocyte diameter or development. In late April, four of six females given the 80/95 GnRHa pellet combination spawned within 9 days. Three fish produced fertile eggs (54 ± 18%). one spawned overripe eggs, and the remaining two increased oocyte diameter and maturation. Three corresponding controls did not spawn, and two of these showed clear signs of atresia within 11 days. In early May, some females were undergoing early FOM and were mature enough to be spawned by hCG injection. Three were given a single 80% CH GnRHa pellet and spawned within 6 days of treatment to produce fertile eggs (44 ± 6%). Of two other females given dual 80% CH GnRHa pellets, one spawned infertile eggs and the other failed to spawn within 9 days. GnRHa implants show promise as a technique for inducing spawning of captive striped bass broodstock although the optimum hormone delivery systems, dosages and release rates should be verified for fish at specific maturational stages.     

Hormone-induced ovulation and spawning of captive and wild broodfish of the catadromous Australian bass, Macquaria novemaculeata (Steindachner), (Percichthyidae)

Australian bass, Macquaria novemacideata (Steindachner),mature but do not spawn in fresh or brackish water ponds. Ovulation and spawning of captive (n = 158)and wild Australian bass (n = 123) was induced in the normal breeding season by single injections of human chorionic gonadotropin (hCG). Doses of 100-4000IU kg-“ hCG induced ovulation and the optimum dosage was 500 IU kg” hCG. The breeding season was from mid-May to late August for wild fish,and extended into September for captive fish. There was a tendency for mean fertilization and hatching success to decline over the breeding season. Greater fertilization and hatching success was obtained from fish which spawned naturally than from stripped fish. Fish spawned after 34,2 ± 0.4 h (mean ± SE, n = 74). Ovulating fish that failed to spawn were stripped after 40.2 ± 0.3 h (n = 76). The timing of stripping and fertilization was an important factor determining hatching success. There was no apparent difference in latency periods or the number of eggs spawned between captive and wild fish. However, the mean number of eggs obtained from naturally spawned fish was higher than for stripped fish. The techniques described in this paper will assist the largescale production of Australian bass by increasing the quality and quantity of larvae from hCG-induced spawnings.     

A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone analogue for ovulation induction in black sea bass Centropristis striata (Linnaeus, 1758)

Mature black sea bass, Centropristis striata L. (200–800 g), were captured in coastal South Carolina during the spawning season and administered hormones for ovulation induction and strip spawning. During both study years, control groups of females were incorporated into the study design and administered sham injections containing physiological saline solution. In 2004, females received a single intramuscular injection of human chorionic gonadotropin (hCG) (330 IU kg⁻¹) (n=8) or two injections of hCG at 24‐h intervals (n=8). In 2005, females received a single injection of hCG (n=10) or an analogue of luteinizing hormone releasing hormone (LHRHa) (n=10). In 2004, all fish administered a single dose of hCG ovulated at least once. Six fish ovulated on two consecutive days and one fish ovulated on 3 days consecutively. In contrast, six of eight fish receiving two doses of hCG ovulated once, five ovulated on 2 days successively and three fish ovulated 3 days in succession. Of the fish that spawned, no differences were found in any reproductive parameters. In 2005, all fish administered hCG or LHRHa ovulated at least once. Three fish administered hCG ovulated twice, four fish ovulated on three consecutive days and one fish 4 days successively. All fish administered LHRHa spawned at least twice, six fish ovulated thrice and three fish ovulated 4 days, successively. A significant difference in fertility was found between hCG (75.6±11.4%) and LHRHa (55.6±27.4%). The results of this study indicate that both hCG and LHRHa are effective for ovulation induction in prespawning black sea bass.     

Induced spawning of hatchery-raised Brazilian catfish, cachara Pseudoplatystoma fasciatum (Linnaeus, 1766)

In the months of January 2001 and 2002, female cachara Pseudoplatystoma fasciatum were selected during their first and second gonadal maturation (2 years and 7 months old and 3 years and 7 months old, respectively) with an of oocyte diameter of 937.5 μm (82.5% with central nuclei and 17.5% with peripheral nuclei). Nine females in first maturation received two doses of carp pituitary extract (CPE), 0.5 mg/kg and 5.0 mg/kg; seven received two doses of human chorionic gonadotropin (hCG), 5 and 10 IU/g; five received doses of 0.5 CPE mg/kg and 5 hCG IU/g (CPE+hCG); and four received 0.9% saline (saline). Nine females from CPE and seven from hCG presented oocytes with the same diameter at the moment of oocyte release (100% with germinal vesicle breakdown and fertilization rate of 53.44±18.3 and 54.81±11.8%; larvae number of 165,330±94.1 and 158,570±20.6, respectively). The five females from CPE+hCG did not respond to the hormonal treatment. The four females from the saline group did not ovulate. In January 2002, 6 of 15 selected females that were going through the second reproductive cycle received CPE (five received hCG and four received saline), showing oocyte diameters similar to the ones in the first maturation. At stripping, CPE females had an oocyte diameter of 1062.5 μm (the hCG females had oocyte diameters ranging from 937.5 to 1125.0 μm; fertilization rates of 56.08±30.9 and 81.90±17.3%; 364,547±244 and 633,129±190, larvae, respectively). The fertilization rates and larvae number were higher in the second gonad maturation, both for CPE and hCG.     

High‐quality spontaneous spawning in greater amberjack (Seriola dumerili,Risso 1810) and its comparison with GnRHa implants or injections

Production of sufficient high‐quality eggs of greater amberjack (Seriola dumerili) still constitutes the main bottleneck for commercial production of this species. The main objective of this study was to compare the quality of spontaneous spawn of greater amberjack with those obtained by either GnRHa injection or GnRHa implant protocols. Captive amberjack broodstock were distributed in three circular tanks of 40 m³. Broodstock from Tank 1 were not hormonally induced and spawned spontaneously, whereas those of Tank 2 were intramuscularly injected with GnRHa (20 µg/kg body weight) and those of Tank 3 were given EV‐500 µg GnRHa implants. The number of eggs per spawn obtained in the broodstock without hormonal treatment was larger than in those obtained with injections or implants. Egg quality was best in broodstock with spontaneous spawn, followed by GnRHa‐injected fish and then GnRHa implants. Besides, size of larvae from control and injected broodstock was similar between them and significantly higher (p < 0.01) than those from GnRHa implant spawn. Overall, this study showed that it is possible to obtain very high‐quality spontaneous spawn in greater amberjack, providing the adequate conditions. Furthermore, GnRHa weekly injections lead to similar egg viability and hatching rates than spontaneous spawn and higher fertilization rates than GnRHa hormonal implants, which is better than in previous studies.     

Induced Spawning and Egg Quality Evaluation of Red Snapper, Lutjanus campechanus

Brood red snapper, Lutjanus campechanus, were captured from the wild and induced to ovulate by human chorionic gonadotropin (hCG) injection; eggs and sperm were manually stripped; and the eggs were artificially fertilized. Eggs were evaluated by the following egg quality parameters: buoyancy, fertilization, egg size, and oil globule size and number. The relationship of these egg quality parameters and brood characteristics (female size, fecundity, time of year, and response time to ovulation) to 36 h posthatch larval survival was considered. Injection of wild‐caught red snapper females with 1100 IU/kg of hCG resulted in 75% of the females ovulating. The average fecundity was 343,377 ± 30,805 eggs/kg, with a mean percent fertilization of 79.0 ± 1.74%. The mean percentage of floating eggs per spawn was 91.8 ± 1.75%. Mean egg diameter for floating eggs was 778.3 ± 2.09 μm, with a mean oil globule diameter of 117.5 ± 1.53 μm. Brood‐related characteristics were a better predictor of larval survival than postovulation egg characteristics. The percentage of floating eggs in a spawn was not correlated to larval survival. Spawns with eggs having a single oil globule had a similar larval survival as those eggs where multiple globules were common. No clear relationships were found for any one factor and larval survival, but rather a combination of factors was more predictive of survival, most notably spawn date, fecundity, and response time following hCG injection.     

Induced Spawning of Nassau Grouper Epinephelus striatus

Nassau grouper Epinephelus striurus females ovulated 48–51 h after the first of two intramuscular injections of human chorionic gonadotropin given 24 h apart (usually 0.7 IU/gram body weight). Typical spawns contained 400,000–600,000 eggs. With fresh milt and clean water, fertilization rate was 85 and 86%. Survival from fertilization to first feeding for six spawns was 73–94%.     

Effect of Luteinising Hormone-Releasing Hormone Analogue and Human Chorionic Gonadotropin on Ovulation, Plasma and Ovarian Levels of Gonadal Steroids in Greenback Flounder Rhombosolea tapirina

Female greenback flounder Rhombosolea tapirina were treated with either des Gly¹⁰ [D-Ala⁶] LHRH ethylamide (LHRH-a) at 50 or 100 pg/kg intraperitoneal injection (ipi), LHRH-a cholesterol pellet (LHRH-a pellet) 100 μg/kg implanted intraperitoneally, or human chorionic gonadotropin (hCG) 1,000 IU/kg ipi. Treatment with hCG, LHRH-a (50 μg/kg) ipi and LHRH-a pellet increased the total number of ovulations above control levels. LHRH-a (100 μg/kg) ipi was not consistently more effective than the control and both LHRH-a (50 μg/kg) ipi and LHRH-a pellet induced more ovulations than LHRH-a (100 μg/kg) ipi. Of those fish that ovulated, most ovulated more than twice, and most ovulations occurred at daily intervals. Oocyte diameters increased and oocyte stage advanced significantly in response to all exogenous hormone treatments. This was accompanied by increases in plasma and ovarian levels of 17β-estradiol (E₂, and in most cases, plasma and ovarian levels of testosterone (T). Plasma and ovarian levels of 17,20β-dihydroxy-4-pregnen-3-one (17,20βP) were not consistently elevated in association with reproductive events.     

Spawning induction in Kutum Rutilus frisii kutum (Kamenskii, 1901) using carp pituitary extract or GnRH analogue combined with metoclopramide

Kutum Rutilus frisii kutum (Kamenskii, 1901), Cyprinidae is an endemic fish of the Caspian Sea. Iranian Fisheries Organization (Shilat) produce up to 200 million fry (1–2 g body weight (b.w.)) to restock the Caspian Sea population annually. Some of these fry are produced by spawning induction in broodfish by carp pituitary extract (CPE). The objective of this study was to assay the effectiveness of the gonadotropin releasing hormone analogue (d ‐Ala⁶, Pro⁹‐Net GnRH) alone or in combination with metoclopramide (MET), a dopamine antagonist, on the percentage of ovulated females, latency period, ovulation index and fertilization success. The following hormone treatments were tested: single injection of 2 mg kg^?1 b.w. of CPE as a positive control, GnRHa alone 20 and 40 μg kg^?1 b.w. and combination of GnRHa and MET as follows: 5 μg+2.5 mg, 10 μg+ 5 mg and 20 μg+10 mg kg^?1 b.w. Negative control group was injected with 0.7% saline. The percentage of ovulated females, ovulation index and fertilization success were 90%, 71.3±1.24%, 68.4±2.3%, respectively, in the group treated with GnRHa+MET at a dose of 20 μg+10 mg kg^?1 b.w. and were significantly higher than those in the positive control (60%, 64.5±0.23%, 69.1±4.5%) (P<0.05). However, the latency period in this group was longer than that in the positive control (P<0.05). Only 20% and 40% fish ovulated in groups that received 20 or 40 μg kg^?1 b.w. GnRHa. No fish ovulated in the negative control.     

The use of luteinizing hormone releasing hormone analogue for ovulation induction in black sea bass (Centropristis striata)

Interest in commercial production of black sea bass has increased in recent years, but reliable spawning methods remain problematic. The objective of this study was to evaluate the effects of oocyte size and luteinizing hormone releasing hormone analogue (LHRHa) dosage and delivery systems on ovulatory success for in vitro fertilization. Vitellogenic females with maximum oocyte diameters of 400–625 μm were implanted with a 95% cholesterol–5% cellulose pellet containing 50 μg of LHRHa. Fish with maximum oocyte diameters < 450 μm failed to ovulate. In contrast, 90% of fish with 500 μm oocytes spawned within 36 h and 40% of this group ovulated a second time. All of the females containing oocytes > 550 μm ovulated. In a second experiment, females with uniformly vitellogenic oocytes (> 500 μm) and implanted with 50 μg LHRHa ovulated substantial numbers of eggs (45,000–192,000 eggs/kg body weight (BW), but fertility was consistently low (0–15%). In a third experiment, 19 of 39 females receiving implants containing 6.3–23.6 μg LHRHa/kg BW during the spawning season ovulated, but fecundity (17,000–339,000 eggs/kg) and fertilization (0–98%) were highly variable. When fish were grouped by developmental index, calculated as the number of oocytes with diameters > 400 μm/total number of oocytes measured, there were no statistical differences among groups with respect to the number of spawns, fecundity or fertilization success. In a fourth experiment, 11 of 13 females with a clutch of fully vitellogenic oocytes that were injected with 20 or 100 μg/kg BW LHRHa ovulated between 1 and 2 times on consecutive days. Five of seven given an implant containing 12.5 μg LHRHa ovulated one or more times. Fish implanted with shams or injected with vehicle alone did not ovulate in any of the experiments. No differences were found in the number of spawns, fecundity or fertilization success from fish receiving different doses of injected or implanted LHRHa. Incubation of pooled eggs produced 155,000 larvae (60% hatch) and 95,000 one-gram juveniles. These results demonstrate that injected or implanted LHRHa is effective for inducing ovulation in black sea bass with maximum oocyte diameters > 500 μm.     

Dopaminergic influence on gonadotropin secretion in white sturgeon (Acipenser transmontanus)

The effects of dopamine on the secretion of two sturgeon gonadotropins (stGTH I and stGTH II) in sexually mature male white sturgeon (Acipenser transmontanus) were evaluated. In Experiment I, sturgeon were given intraperitoneal injections of physiological saline (PS), dopamine (100 mg kg⁻¹), the gonadotropin releasing hormone analog D-Ala⁶-des-Gly¹⁰-GnRH ethylamide (GnRHA) (10 μg/kg⁻¹), and a combination of GnRHa and dopamine. Fish receiving only GnRHa had significantly higher (p < 0.05) concentrations of plasma stGTH I and stGTH II compared to fish receiving PS, dopamine, or a combination of GnRHa and dopamine. Two hours following its administration, dopamine was effective in decreasing plasma concentrations of both stGTHs that were previously elevated by GnRHa. Dopamine or PS administered by themselves did not alter plasma concentrations of either stGTH. In Experiment II, sturgeon injected intraperitoneally with a combination of GnRHa and pimozide had significantly higher (p < 0.05) concentrations of plasma stGTH I and stGTH II compared to males receiving GnRHa or pimozide alone. While this effect of GnRHAa + pimozide was observed in the spring, no such potentiation was seen in these fish during the summer (Experiment III). These results represent the first evidence of dopaminergic inhibition of GnRH-induced pituitary gonadotropin secretion in Chondrostean fish.     

Induced Spawning of Wild American Shad Alosa sapidissima Using Sustained Administration of Gonadotropin-Releasing Hormone Analog (GnRHa)

American shad Alosa supidissima broodstock were collected from the Susquehanna River during their spawning migration. Mean volume of expressible milt (± standard deviation) was 2.5 (±1.7) mL/kg body weight; mean spermatozoid count was 66.2 ± 10⁹ (±163 ± 10⁹) spermntozoa/mL; and duration of 50% motility was 36.5 (±10.3) see. Ovarian biopsies indicated the presence of oocytes of various sizes (200–2,000 μm in diameter) and stages of development. Fish were implanted with a delivery system loaded with gonadotropin-releasing hormone analog (GnRHa) and started spawning 2 d after treatment. Fertile eggs were collected daily for the next 9 d, for a total of 50,100 eggs/kg body weight with a mean fertilization success of 62%. Upon cessation of spawning, the ovaries of all females still contained large numbers of oocytes at various stages of development, as at the beginning of the experiment, but with a greater number of atretic oacytes. Our observations show that American shad have an asynchronous ovarian development, and treatment with a GnRHa delivery system is effective in inducing several successive spawns of fertile eggs.     

Artificial induction of maturation and fertilization in the Japanese eel, Anguilla japonica

Repeated injections of salmon pituitary extract (20 mg per fish per week) induced vitellogenesis in feminized, cultivated Japanese eels (Anguilla japonica). Oocytes were attained at the migratory nucleus stage after 11 or 12 injections. Addition of 17,20-dihydroxy-4-pregnen-3-one (DHP) into the incubation medium induced germinal vesicle breakdown (GVBD) in the oocytes at the migratory nucleus stage. An injection of DHP (2 µg g^-1 BW), given 24h after an injection of salmon pituitary extract (20 mg fish^-1), succeeded in inducing maturation and ovulation in females which contained occytes at the migratory nucleus stage. Most fish ovulated 15–18h following the DHP injection. Eggs that were ovulated within 15h after the DHP injection showed high fertility and hatchability, but eggs ovulated 18 or 21h after the DHP injection, showed considerably lower fertility and hatchability. A delay between ovulation and stripping of the eggs rapidly decreased both the fertility and hatchability within 6–9h after ovulation, indicating that artificial fertilization must be carried out immediately after ovulation. Repeated injections of human chorionic gonadotropin (hCG) at a concentration of 1 IU g^-1 BW week^-1 induced spermatogenesis, spermiation, and the acquisition of potential for sperm motility in cultivated males. Most males spermiated after the fifth or sixth injection of hCG, and the milt weight gradually increased and remained constant (1–2 g) from the 11th to 31th injection. Sperm motility peaked 24h after each weekly injection, and decreased from the 3^rd day after the injection. Potassium ions are an essential constituent for the maintenance of motility in the eel spermatozoa. Artificial seminal plasma containing 15.2 mM KCl is applicable as a milt diluent. Using these techniques developed for female and male eels, we have succeeded in obtaining many fertilized eggs from cultivated eels.     

Induced ovulation using LHRHa in anadromous obscure puffer Takifugu obscurus cultured entirely in freshwater

Puffer fishes of the order Tetradontidae exist in both anadromous and non-anadromous seawater resident forms. The obscure puffer akifugu obscurus is an anadromous species whose intensive culture in freshwater has developed rapidly in China. To mass produce larvae for intensive culture, induction of ovulation of 3-year-old obscure puffer cultured entirely in freshwater was attempted by giving the fish multiple injections of LHRHa. Twenty 3-yearold cultured obscure puffers were treated with multiple injections of LHRHa at a constant dosage of 30 g of LHRHa kg-1 of body weight with 36 h interval between injections. Beginning two days after hormonal treatment, abdominal palpation was performed every day to check expansion and hardening of the abdomen of the fish due to hydration of oocytes that indicates the completion of final oocyte maturation. The first four fish ovulated after the second LHRHa injection on day 3, but most females ovulated after the fourth hormone injection. A total of 18 fish (90%) ovulated over a period of 7 days, while no fish ovulated in the control group. The mean fertilization rate and the mean hatch rate was 67.1% and 87.6%, respectively. The viability of newly hatched larvae was very good, with a high survival rate of 98.6% 24 h post-hatch. These results indicate that cultured obscure puffer, which are not allowed to undergo diadromous migration and are entirely cultured in freshwater, could be induced to complete maturation and ovulate by simple multiple injections of LHRHa and that the eggs could be successfully hatched into viable larvae.     

异齿裂腹鱼人工规模化繁殖技术研究

2010年4～6月,对野生异齿裂腹鱼(Schizothorax o' connori)人工规模化繁殖技术进行研究,并初步进行产后亲鱼恢复培养技术研究.对108尾雌鱼进行干法人工授精,共采卵104万多粒,孵出仔鱼62万多尾.其中45尾雌亲鱼自然成熟,共采卵46.8万多粒,平均受精率和孵化率低于人工催产雌鱼卵.人工催产83...     

Induced maturation and spawning of domestic and wild striped bass, Morone saxatilis (Walbaum), broodstock with implanted GnRH analogue and injected hCG

Abstract. A domestic striped bass. Morone saxatilis (Walbaum), broodstock was established by rearing fish to sexual maturity in ponds. A method was developed to reproduce the domestic females, and also wild females too immature to be successfully induced to spawn with injected human chorionic gonadotropin (hCG). The fish were implanted with pellets containing 100–150μg of a synthetic analogue of mammalian gonadotropin reieasing-hormone, [D-Ala⁶- Pro⁹-NEt]-LHRH (mGnRHa), in a matrix of cholesterol (CH) and cellulose. They were implanted with one fast hormone-release (80% CH) pellet and one slow hormone-release (95% CH) pellet and allowed to mature for 1–3 days, until they entered the process of final oocyte maturation and were induced to ovulate or spawn with an hCG injection. The secondary hCG injection was found to be necessary to speed the maturation of the wild fish; they otherwise would succumb to the stresses of capture, handling and confinement before they could be spawned. The total mGnRHa dosages used ranged from 33 to 111μg mGnRHa/kg body weight, and the hCG doses were either 165 or 330 IU hCG/kg body weight. Using the combined mGnRHa implant-hCG injection technique, fry production rates were comparable to those obtained using fully mature wild females taken directly from their spawning grounds.     

Induced Final Maturation and Spawning of the Marbled Grouper Epinephelus microdon Captured from Spawning Aggregations in the Republic of Palau, Micronesia

A high percentage (98.3%, N = 60) of the marbled grouper Epinephelus microdon individuals captured from spawning aggregations during July and August 1993 in the waters surrounding the island of Koror, Republic of Palau, Micronesia, were in the stage of maturity at which final maturation and spawning could be hormonally induced. The sex ratio of the captured fish was highly skewed towards males (4 male:1 female). Sexually immature females comprised the smallest size class, (<0.6 kg body weight (BW) or 33.0 cm total length (TL)), while sexually mature females were restricted to the 0.6–1.5 kg BW (33.0–46.4 cm TL) groups. Males predominated in size classes >0.6 kg BW, and individuals >1.5 kg BW (46.4 cm TL) were exclusively male. All females with oocytes that averaged ( N = 50) >400 μm in diameter were successfully induced to spawn by a two-injection protocol using human chorionic gonadotropin (HCG) at total dosages of 2,100–3,200 IU/kg fish. All males used in the spawning trials were administered a single injection of HCG at dosages of 500 or 1,000 IU/kg fish. Fecundity ranged between 7.96 × 10⁵−1.24 × 10⁶kg BW, average spawned egg diameters ranged between 769–832 μm, percent fertilization ranged between 32.6%–99.9%, and hatching percentages were >90.0%. Total fat content of eggs obtained from a pooled spawning event was 14.1 mg/100 mg dry weight. The data indicate that HCG is a suitable treatment for the induction of spawning in marbled grouper females that possess a mean oocyte diameter of 400 μm or greater.     

Effects of emulsified versus saline administration of GnRHa on induction of ovulation in rainbow trout, Oncorhynchus mykiss

The effects of saline-dissolved or Freund's incomplete adjuvant (FIA)-emulsified GnRHa treatment on the induction of ovulation in rainbow trout, Oncorhynchus mykiss, were examined. The FIA-emulsified GnRHa was first diluted in 0.25 ml physiological saline and then mixed with an equal volume of FIA. Fish were selected in the beginning of the spawning season and were allocated into four groups and were treated intraperitoneally with (a) 0.5 ml of emulsified GnRHa (GnRHa–FIA), (b) 0.5 ml saline-dissolved GnRHa in a single injection (GnRHa-1), (c) 0.5 ml saline-dissolved GnRHa in two injections spaced 1 d apart (GnRHa-2) and (d) 0.5 ml of saline (Control). The GnRHa dose in all hormone treatments was 25 µg kg^− 1. All fish in the FIA–GnRHa and GnRHa-2 groups ovulated within 10 and 11 d after treatment, respectively. In contrast, only 75% in the Control fish and 60% of the fish in the GnRHa-1 group ovulated within 36 d after treatment. None of the treatments caused any pre- or post-spawning mortality in the broodstock. Fertilization, eyeing and hatching percentages of the produced progeny were normal in all the treatment groups and did not differ significantly among them. In conclusion, FIA-emulsified GnRHa can be effective in advancing the onset of and synchronizing the ovulation of rainbow trout within a two-week period, thus shortening the egg collection period, without affecting broodstock survival and egg quality.