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.     

Induced final maturation and ovulation in a small anabantoid teleost, the dwarf gourami, Colisa lalia. I. The effects of gonadotrophic preparations, LHRHa, steroids and thyroid hormones

Colisa lalia (Ham.) has been used as a model for the development of techniques for induced spawning that are applicable to small teleosts where ovulation requires prolonged exposure to suitable breeding conditions.Human chorionic gonadotrophin (hCG; 4 IU per fish) induced ovulation within 24 hours, whereas homologous pituitary extracts were relatively ineffective. When administered in saline ( 20 g per fish per injection), des-Gly10,[D-Ala6]- LHRH N-ethylamide (LHRHa) was ineffective, but it stimulated ovulation in a proportion of fish when administered ( 1.6 g per fish) as an emulsion in Freund's incomplete adjuvant (FIA). Together, these results suggest that ovulation requires the synthesis as well as the secretion of gonadotrophin in C. lalia.Long-term treatment with thyroid hormones appeared to enhance the ovulatory response to LHRHa in FIA, possibly by effects on the ovary; whereas the various steroids tested were ineffective at the dosages used     

多巴胺拮抗物 pimozide(PIM)或排除剂 reserpine(RES)和促黄体素释放激素类似物(LHRH-A)诱导养殖鱼类促性腺激素分泌和排卵的研究

由多巴胺拮抗物PIM或排除剂RES组成的高效新型鱼类催产合剂能有效的诱导主要的淡水养殖鱼类促性腺激素分泌和产卵，其中青鱼催产率为80％，鲢鱼为84％，鳙鱼和草鱼为88％，长春鳊为94％，鲮鱼为96％。试验表明这种催产剂的主要优点是：催产效果高而稳定，可以完全取代鱼脑垂体；对当年第二次成熟和产卵的亲鱼同样有效；有利于保护亲鱼；节省劳力和成本。     

Induced spawning in perch, Perca fluviatilis L. using carp pituitary extract and HCG

Induced spawning in Eurasian perch, Perca fluviatilis L., was studied using human chorionic gonadotrophs (hCG) or acetone-dried common carp pituitary with or without hCG. The dose administered to fish was 5700 IU hCG kg^-1 or 4.0 mg kg^-1 carp pituitary with or without 500-700 IU hCG kg^-1 for females and 2850 IU hCG kg^-1 or 2.0 mg kg^-1 carp pituitary with or without 250-350 IU hCG kg^-1 for males. There were no statistically significant differences in quantity of milt in treated and control groups, although the best result was observed when males were treated with a triple injection of hCG and carp pituitary extract. Male spermiating success, expressed as a quantity of milt, was negatively correlated with fish weight. All females treated in this experiment had oocytes at the same division, so time of ovulation was highly synchronous. Spawning success, expressed as a spawning effectivity coefficient (S_c), was highest in fish treated with the triple injection. Spawning methods described in this paper were successful, even though hormones from another order of fish were used.     

Time course of final oocyte maturation and ovulation in chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> induced by hCG and GnRHa

Abstract:   The question of whether the ovulation and spawning time in chub mackerel Scomber japonicus is entrained by a circadian rhythm was raised by our previous experiments. Further questions were also raised about whether the time course of human chorionic gonadotropin (hCG)-induced final oocyte maturation (FOM) and ovulation reflected the natural time course induced by endogeneous pituitary gonadotropin (GtH). To address these questions, hCG and gonadotropin-releasing hormone analog (GnRHa) were administered at two 'opposite' times, 14:00 and 02:00 hours, and the time courses of FOM and ovulation were compared. When hCG was injected, ovulation occurred 33 h post-injection in both groups, regardless of the timing of the hCG injection. The timing of ovulation in chub mackerel depends on the timing of hCG injection, but apparently not on circadian rhythms. When GnRHa was injected, ovulation began at 36 h post-injection of GnRHa, regardless of the timing of injection. These results indicate that the time course of FOM and ovulation in the chub mackerel followed a similar pattern whether stimulated by hCG injection or spontaneous luteinizing hormone (LH) surge because GnRHa induces the secretion of endogenous GtH (primarily LH) from the fish pituitary. Thus, it is concluded that the time course of hCG-induced FOM and ovulation in chub mackerel follows the natural time course induced by endogenous pituitary LH.     

红鳍东方鲀的人工繁殖试验

采用网箱养殖四年的红鳍东方鲀经过颗粒状的促黄体生成激素释放素(LH-RH)的植埋,促使性腺成熟,再通过注射绒毛膜促性腺激素(HCG)与大麻哈鱼脑垂体混合液诱导排卵,采卵后进行人工授精,得到红鳍东方鲀受精卵,孵化后得到红鳍东方鲀仔鱼。     

Induced spawning in bream, Abramis brama (L.), using carp and bream pituitary extract and hCG

Induced spawning in bream, Abramis brama (L), was studied using acetone-dried common carp pituitary (CP) and bream pituitary (BP) with or without the addition of human chorionic gonadotrophin (hCG). The total dose administered to fish was of 5.0 mg kg^?1 BP or 4.0 mg kg^?1 CP with or without 2000-2200 IU hCG kg^?1 for females and 2.5 mg kg^?1 BP or 2.0 mg kg^?1 CP with or without 1000–1100 IU HCG kg^?1 for males. In all male treated groups 100% of spermiation was observed: in females the most effective method was a triple injection with hCG and carp pituitary, resulting in 79% of females ovulated (over 68% of eyed eggs). Biological quality of eggs, expressed as a percentage of eyed eggs, was negatively correlated with time elapsing between resolving (final) injection and ovulation. Spawning success, expressed as a value of S_e (spawning effectiveness coefficient), was higher in fish treated with triple injection.     

Utilization of cocoa-pod husk in low-cost diets by the clariid catfish, Clarias isheriensis Sydenham

Abstract. Clarias isheriensis Sydenham fingerlings (60·42 ± 1·13g) were fed for 180 days with pelleted diets in which maize was progressively replaced with graded levels (0, 15,30,45%) of cocoa-pod husk meal (CPHM). There was a decline in growth rate with increase in dietary CPHM level, though this was not significantly different (P < 0·05). Differences in feed conversion, nutrient digestibility and carcass characteristics among different dietary treatments were also not significant (P > 0·05). Fish mortality was low (< 2%) in all dietary treatments tested. Lack of significant difference in growth performance and nutrient utilization indicated that CPHM could be incorporated into low-cost diets for C. isheriensis up to the 30% level.
In a second experiment, three feeding regimes of 15% CPHM diet were tested with C. isheriensis fingerlings (60·81 ± 1·87g) over 180 days and results showed no significant (P > 0·05) differences among the feeding regimes tested, with regards to growth, feed conversion and fish yield. This suggests that the growth of C. isheriensis from fingerling to adult size (≥ 200g) can be satisfied in one single feeding equivalent to 3% body weight per day.     

Induction of ovulation in ornamental common carp (Koi, Cyprinus carpio L.) using LHRHa ([d-Ser(tBu)6, Pro9-NEt]-LHRH) combined with haloperidol and carp pituitary extract

The effects of a single injection of mammalian superactive analogue [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa (20 μg/ kg^?1) combined with the dopamine antagonist, haloperidol (HAL, 0.5 mg kg^?1), for induction of ovulation in the koi carp broodstocks were determined under routine hatchery conditions. The results were compared with classic carp pituitary extract (CPE, double injection) application (water temperature 22 °C). Physiological saline (0.7% NaCl)‐injected fish were used as a control group and no ovulation occurred in this group. The spawning ratio was high in the LHRHa+HAL treatment group and in the CPE treatment group (6/7 and 5/7 respectively). The latency period was 14–16 h in the LHRHa+HAL treatment group and 12–14 h in the CPE treatment group (after the second injection). There was no difference between the two ovulating groups with respect to the spawning index (the weight of eggs as a percentage of female body weight) and fertilization rate of eggs (P>0.05). As a result, ovulation can be induced successfully in koi carp broodstocks with 20 μg kg^?1 [d ‐Ser(tBu)⁶,Pro⁹‐NEt]‐LHRHa+0.5 mg kg^?1 HAL treatment in a single injection without decreasing the egg quality. Application of this combination can be useful for hatchery and broodstock management in koi carp culture.     

Controlled reproduction in Anguilla anguilla (L.): comparison between spontaneous spawning and stripping‐insemination approaches

This study aimed to compare the fertility of eggs between artificially matured female silver eels that spawned spontaneously and those that were spawned by manual stripping. The effects of the two methods of spawning on ovulation and fertilization rate were also investigated. For this purpose, 18 wild female European eels captured in Bonello lagoon (North Adriatic Sea) were carp pituitary extract‐injected to undergo sexual maturation and ovulation; a final injection of 17,20β‐dihydroxy‐4‐pregnen‐3‐one (DHP) was administered when at least 30% of the oocytes were fully transparent. After the DHP‐injection, nine eels were transferred to a new closed recirculating aquaculture system, where they were housed with spermiating males (sex ratio 4/1) to allow spontaneous spawning (SPT‐group); the remaining nine eels were transferred to a 250 L tank and ovulation was checked at four‐hourly intervals by manual stripping (STR‐group). The number of eggs per female in the SPT‐group was significantly greater than that in the STR‐group. Furthermore, fertilization rates in the SPT‐group were notably higher than those observed in the STR‐group. Significantly, the best performances were obtained among eels in which at least 50% of oocytes were fully transparent at the time DHP was administered. We conclude that the fertility of eggs from spontaneously spawning eels is superior to that of eggs acquired by strip‐spawning and artificial fertilization.     

In vivo activity of native GnRHs and their analogues on ovulation in the African catfish, Clarias gariepinus (Burchell)

Four distinct forms of native gonadotropin‐releasing hormone (GnRH) and two newly designed analogues were tested for their in vivo activity to induce ovulation in African catfish. The effects of these peptides on ovulatory parameters were compared with those of carp pituitary and [d ‐Ala⁶, Pro⁹‐NEt]‐mammalian GnRH analogue (mGnRHa), two tested ovulation‐inducing agents in African catfish. Assessment of ovulation was carried out by determining the ovulation ratio and the relative quantity of egg produced. From the results of the experiments, the order of potency of the native GnRH peptides is summarized as chicken GnRH‐II (cGnRH‐II) >salmon GnRH (sGnRH) >mammalian GnRH >chicken GnRH‐I (cGnRH‐I). Chicken GnRH‐II was as potent as mGnRHa while cGnRH‐I was totally ineffective. The new d ‐Orn⁶‐cGnRH‐II and d ‐Orn⁶‐sGnRH with a substitution at position 6 with d ‐isomer residue were as potent as the most extensively used mGnRHa, indicating that the position 6 modification might be more crucial than the substitution at the C‐terminal. On the basis of our results, the potential use and incorporation of cGnRH‐II and sGnRH for the development of more generic spawning induction therapies are suggested.     

Artificial spawning of common tench <Emphasis Type="Italic">Tinca tinca</Emphasis> (Linnaeus, 1758), obtained from wild and domestic stocks

The study encompasses three reproduction seasons. Tench spawners caught during the spawning season originated from carp ponds (domestic stock) and a lake (wild stock). Fish were reproduced under controlled conditions after hormonal stimulation with GnRHa-containing pellets combined with metoclopramide (Ovopel) or carp pituitary homogenate (CPH). As a result of hormonal stimulation, eggs were obtained from a larger number of females originating from the lake (71.7%) than those originating from the pond (58.3%), although no other statistical differences were found. A similar relationship was recorded for the spermatozoa motilities (range from 72 to 76%). The obtained results indicate that both investigated reservoirs are suitable for tench broodstock management due to the fact that synchronization of ovulation among different stocks is easy to achieve. For this purpose, among the tested spawning agents, Ovopel could be recommended as being slightly more effective.     

Ovulation induction in northern pike Esox lucius L. using different GnRH analogues, Ovaprim, Dagin and carp pituitary

The traditional GnRH analogue treatments applying the chemicals in pure form proved to be ineffective in inducing ovulation in northern pike (Esox lucius L). Neither mGnRHa ([D‐Ala⁶, Pro⁹NEt]‐mGnRH) nor sGnRHa ([D‐Arg⁶, Pro⁹‐Net]‐sGnRH) administered alone or together with pimozide (mGnRHa), metoclopramide (mGnRHa) or domperidone (sGnRHa) induced ovulation in females, whereas in groups receiving a carp pituitary injection most females ovulated. Spawning‐inducing agent Dagin did not induce ovulation, whereas all but one female ovulated in the carp pituitary‐treated group. Treatment with another preparation, Ovaprim, resulted in similar or lower ovulation ratio than treatment with carp pituitary. After the Ovaprim treatment, time to ovulation was not as predictable as after the carp pituitary injection. The mean fertilization rate was relatively low and similar in the groups treated with Ovaprim (54.7 ± 12.3% and 58.7 ± 19.1% for the first and second experiment respectively) and with carp pituitary (53.7 ± 10.5% and 58.9 ± 14.9%). The mean pseudogonadosomatic index (PGSI) was also similar between the Ovaprim‐treated group (14.5 ± 6.1%) and the carp pituitary‐treated one (17.9 ± 4.1%). In the present experiments, treatment with Ovaprim was less effective than that with carp pituitary.     

Preliminary data on the induction of ovulation in white sturgeon (Acipenser transmontanus Richardson)

Forty-six white sturgeon females captured prior to their spawning migration in the San Francisco Bay and during their spawning run in the Sacramento River were induced to ovulate with three different hormonal substances: white sturgeon and common carp crude pituitary extracts, and [D-Ala⁶] GnRH analogue. All three inducing agents were potent at the respective doses of 2.5 mg, 4.0 mg and 0.1 mg dry matter per kilogram of female body weight. The best ovulatory responses and egg fertilities were observed in fish administered hormonal treatment during the months of March and April. Fish captured prior to and during their spawning migration exhibited similar spawning success.     

Evaluation of cGnRH IIa for induction spawning of two ornamental Epalzeorhynchos species

The administration of exogenous hormones for induction spawning of captive fish is a common practice throughout the finfish aquaculture industry. Induced spawning protocols for ornamental cyprinids commonly rely on sGnRH IIIa, commercially available as Ovaprim. In this study, the efficacy of an alternative spawning aid, cGnRH IIa, was evaluated relative to Ovaprim, in two commonly cultured ornamental Cyprinids: the Redtail Sharkminnow Epalzeorhynchos bicolor and the Rainbow Shark E. frenatum. Broodstock of each species were injected with either a positive control (0.5 µl/g Ovaprim), a negative control (propylene glycol) or one of three doses of cGnRH IIa (50, 100, or 200 µg/kg). Following spawning aid injection, ovulation success, fecundity, fertilization success, embryo diameter, hatch success and larval notochord length were evaluated. Ovulation success (82 ± 8%) was statistically similar to the positive control group for all experimental doses of cGnRH IIa in E. bicolor, while 50 and 100 µg/kg cGnRH IIa doses resulted in significantly higher ovulation success (100.0% and 83.0% respectively) than the positive control (17.0%) in E. frenatum. All other parameters did not vary significantly among treatments. These results indicate cGnRH IIa can be successfully used as a spawning aid in both species.     

Artificial spawning of carp (Cyprinus carpio L.): differences between females of Polish strain 6 and Hungarian strain W treated with carp pituitary homogenate, Ovopel or Dagin

Investigations of the effects of controlled reproduction were conducted on carp females of Polish strain 6 and Hungarian strain W after ovulation stimulation with carp pituitary homogenate (CPH) (0.3 mg+2.7 mg kg^?1 body weight), Ovopel (1/5+1 pellet kg^?1 body weight) or Dagin (1 dose kg^?1 body weight). In both strains, the highest percentage of spawning females was found after the Ovopel treatment. After treatments with CPH and Dagin this percentage did not differ in strain 6; however, after Dagin, it was higher in fish of strain W. After CPH or Ovopel stimulation the females of line 6 yielded eggs of higher weight compared with line W. After the Dagin treatment the weight of eggs obtained from females, both strains was similar but much lower than after the two remaining agents. The quality of eggs expressed both by the percentage of fertilization and that of living embryos after 24 and 36 h incubation was similar in the two strains only in the group treated with Ovopel. After CPH application eggs of better quality were obtained from strain 6 compared with strain W; however, after the treatment with Ovopel, eggs of much better quality were noted in strain W. In general, the best effects of reproduction were recorded in the case of Ovopel and the poorest in that of Dagin. Females of strain 6 yielded eggs of a higher weight but the percentages of spawning females were higher in strain W after each of the applied spawning agents. A new approach to this problem consisted in investigating the interaction between ovulation stimulators and origin of females of fish treated with Dagin.     

Effect of different commercial spawning agents and thermal regime on the effectiveness of pikeperch, Sander lucioperca (L.), reproduction under controlled conditions

Pikeperch, Sander lucioperca (L.), has been identified as one of the most perspective candidates for diversification of freshwater aquaculture. However, some aspects of production are still being developed, and controlled reproduction is one of the bottlenecks. The aim of the present study was to compare the effectiveness of different commercial spawning agents in the induction of final oocyte maturation (FOM) and ovulation in wild spawners. Within the study, four spawning agents [human chorionic gonadotropin (hCG), mixed human and horse gonadotropin (PG-600), carp pituitary (CPH) and mammalian GnRH analogue combined with metoclopramide (Ovopel)] in different thermal regimes (13 and 15 °C) were tested. In both thermal regimes, the highest (P < 0.05) ovulation rate among the treatment groups was obtained after stimulation with hCG (100 % in both cases). Latency time was the shortest in groups where CPH was used (2–3 and 3–4 days for 15 and 13 °C) and was similar in the remaining groups (3–4 and 4–5 days for 15 and 13 °C, respectively). Embryo survival was the highest in groups treated with hCG (78.9 and 81.3 % at hatching stage for 15 and 13 °C, respectively). Hormonal stimulation did not significantly affect spermiation rate or spermatozoa motility (P > 0.05). Based on the obtained results, hCG can be recommended for induction of FOM and ovulation in pikeperch. In addition, the thermal regime within the tested range seemed to have no effect on the reproduction outcome, and the application of lower temperature only prolonged the time of ovulation.     

Induced Spawning of the Indian Catfish,Heteropneustes fossilis,by GnRH Analogue Alone or in Combination with Dopamine-Affecting Drugs

ABSTRACT

Mature female Indian catfish, Heteropneustes fossilis were induced to spawn in spawning season (July-August) by various ovulation-inducing agents, and the frequency of ovulation was checked after a latency of 12-14 hours of the treatment. Egg mass and fertilization rate were taken as end-points of spawning performance. [D-Ala⁶-Pro⁹] ethylamide GnRH analogue in dosages of 0.075, 0.15, 0.2, and 0.5 μg/g of body weight resulted in 28.5%, 71%, 86%, and 86% ovulation, respectively, but the lower doses of 0.01 and 0.05 μg/g body weight failed to induce ovulation in early spawning phase (July). The dopamine-2 (DA2) receptor antagonists pimozide, domperidone, and metoclopramide potentiated the anovulatory dose of 0.05 μ g GnRH analogue to induce ovulation in 86%, 86%, and 50% of the females, respectively, in the early spawning phase (July). The DA precursor, L-dihydroxyphenylalanine (L-DOPA; 10 μg/g body weight) and the DA agonist bromocriptine (10 μg/g body weight) decreased markedly the ovulatory response of 0.2 μ g GnRH analogue to 29%. The catechol-amine-synthesis inhibitor a-methylparatyrosine produced a mild depressing effect on the ovulatory response of 0.2 μ g GnRH analogue. Egg mass was high in groups that yielded a high ovulatory response (71% and above) except the a-MPT groups. The fertilization rate, however, was not affected, irrespective of the ovulatory response. The results show that the Indian catfish can be induced to spawn by superac-tive GnRH analogue alone in a dosage range of 0.15-0.2 μg/g body weight or in combination with DA2 antagonists, such as pimozide or domperidone, in a very low dosage of 0.05 μg/g.     

Use of Channel Catfish,Ictalurus punctatus,Pituitary as a Spawning Aid: Influence of Seasons and Preparations on Efficacy

Channel catfish, Ictalurus punctatus, pituitary powder (CP) was prepared and administered to determine the effect of pituitary preparation, head selection criteria, and season of collection on the biological potency of this gonadotropic preparation. Pituitaries were harvested from three catfish processing plants throughout the months of December, February, March, and April to investigate the effect of season on CP potency. Gravid female channel catfish were injected intraperitoneally (IP) with priming (2 mg/kg) and resolving doses (8 mg/kg) of CP treatments, luteinizing hormone‐releasing hormone analog (LHRHa), or saline. Pharmaceutical grade Carbopol 971P resin (CAR) was tested as a 1% aqueous dispersion in physiological saline and administered with CP (10 mg/kg) as a IP single injection at a final volume of 2 mL per kg as a slow‐release vehicle. Individual females were hand‐stripped, and eggs were fertilized with freshly harvested blue catfish, Ictalurus furcatus, spermatozoa. Pituitaries processed with either acetone or ethanol/acetone resulted in significantly different ovulation rates of 35 and 73%, respectively (P≤ 0.01). Ovulation success was not significantly different among CP and LHRHa treatment groups. Ovulation rates were not significantly different among treatment groups of CP collected in April (74%) or December (67%), or collected from male (69%), or small individuals (55%). The shortest latency period among CP treatment was from CP collected in December and from males at 39.1 ± 0.7 and 38.5 ± 0.4 h, respectively. The CAR vehicle treatment performed poorly resulting in an average ovulation rate of 19.0%, with 3.3% viable embryos at 36 h post‐fertilization. Pituitaries collected from channel catfish potentially represent a viable spawning aid for the production of hybrid channel × blue catfish.     

Induced Ovulation, Spawning, Egg Incubation, and Hatching of the Cyprinid Fish Labeo victorianus in Captivity

Two spawning inducing agents—Aquaspawn (a rapidly metabolized synthetic decapeptide gonadotropin-releasing hormone [GnRH]) and Dagin ([D-Arg⁶, Pro⁹-NEt]-sGnRH) combined with 20 mg/kg of the water-soluble dopamine receptor antagonist metoclopramide (GnRH + MET)—were tested for their efficacy in stimulating ovulation in Labeo victorianus held in its natural environment and under captive conditions. Successful ovulation, when migratory vesicle oocytes became completely transparent, was obtained with GnRH + MET, while GnRH only caused oocyte clearance up to the highly translucent phase. L. victorianus eggs were non-adhesive, semi-buoyant, and transparent at ovulation. First hatching occurred after 26 h and lasted for 8 h at 24 C. Water temperature was shown to significantly affect spawning latency and incubation time. Thus, L. victorianus could be successfully induced to spawn using a synthetic gonadotropin-releasing hormone coupled with a dopamine antagonist followed by natural fertilization in floating net cages at temperatures between 24 and 27 C.