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1.
The commercial‐scale production of fish by use of artificial (induced) spawning would require reliable, large‐volume sources of sperm. Cryopreservation can be used to preserve and store sperm within commercial and research germplasm repositories, but is limited in its application to aquaculture. Straw volume and cooling chamber size restrict the quantity of sperm that can be frozen, and straws must be filled by hand. In contrast, the dairy industry has refined methods for freezing of bull sperm, including automation of straw filling and the use of large cooling chambers. These methods could be used for commercial‐scale cryopreservation of fish sperm, although application would require testing. To supply sperm in large volumes, bags originally developed for swine semen could be cooled using dairy protocols and used as a container for fish sperm. The current study documented the use of commercial‐scale dairy cryopreservation techniques for the production of hybrids of channel catfish Ictalurus punctatus (female) by blue catfish Ictalurus furcarus. Four cryoprotectants (methanol, dimethyl sulfoxide, dimethyl acetamide, and glycerol) were initially evaluated for use with blue catfish sperm. During May 2000 and March to April 2001, suspensions of blue catfish sperm were cryopreserved with 10% methanol in 0.5‐mL French straws and in commercial swine semen bags (Cochette* bags, IMV International. Minneapolis, Minnesota, USA). Cryopreservation took place at a dairy breeding cooperative, using technology employed for bull semen. Sperm motility before freezing was 26 ± 18% during Year 1 (2000) and 62 ± 30% during 2001. Sperm were thawed at 40 C and used to fertilize the eggs of channel catfish (yielding hybrids). Motility after thawing for sperm frozen in 0.5‐mL straws was 11 ± 10% during 2000 and 50 ± 24% during 2001. Motility after thawing was 41 ± 17% for sperm frozen in swine semen bags in 5‐mL aliquots and 43 ± 10% for sperm frozen in 10‐mL aliquots. Neurulation of eggs fertilized with thawed sperm from straws was 83 ± 13% during 2000 and 54 ± 27% during 2001. Neurulation was 57 ± 24% using sperm frozen in swine semen bags in 5‐mL aliquots and 55 ± 10% using sperm frozen in 10‐mL aliquots. There was no correlation between sperm motility before freezing (in 0.5‐mL straws) and after thawing during 2000 (r= 0.52) or during 2001 (r= 0.49). In addition, there was no correlation between initial motility and neurulation of channel catfish eggs fertilized using thawed sperm during 2000 (r= 0.14) or during 2001 (r= 0.29). Sperm of blue catfish can thus be cryopreserved at a commercial scale using dairy protocols and can be made available for the production of hybrid catfish when viable eggs are available.  相似文献   

2.
Effectiveness and efficiency of frozen sperm on fertilization and hatching success of eggs from silver barb was examined in relation to cryoprotectants, freezing rate and storage period. Sperm was diluted in calcium‐free Hank's balanced salt solution, equilibrated with dimethylsulphoxide (DMSO), propylene glycol, sucrose or methanol at 5%, 10%, 15% or 20% final concentrations, and frozen in 250‐μL straws using a one‐step freezing procedure (1, 5 and 8°C min?1 from 25 to ?40°C). Highest post‐thaw sperm motility was found from a treatment using 10% DMSO and 5°C min?1 (82.2 ± 2.1%), similar to that of 10% DMSO and 8°C min?1 (87.8 ± 3.2%). Post‐thaw motility of sperm frozen at 5 or 8°C min?1 was significantly higher than 1°C min?1. Relative sperm motility declined significantly after 10 months of cryostorage while viability did not change during a 12‐month cryostorage. Average fertilization rates of sperm after 1 and 4 months of storage were 64.5 ± 4.6% and 61.3 ± 3.4%, respectively, similar to those of fresh sperm (69.6–72.3%). Hatching rates of cryopreserved sperm (45.4–51.2%) were similar to those of fresh sperm (51.8–57.8%). This study developed suitable methods for cryopreservation of silver barb sperm that can be used to facilitate hatchery operation.  相似文献   

3.
In this study, cryopreservation feasibility of Persian sturgeon (Acipenser persicus) and the effect of different doses of 2‐hydroxypropyl‐beta‐cyclodextrin on thawed spermatozoa quality (motility duration and motility percentage) were investigated. For freezing, semen of seven male individuals was pooled in equal volumes and diluted with 4°C [Tris‐HCl (100 mM), pH = 8, DMSO 10%] extenders containing 0, 5, 10, 15 mM of HβCD in a ratio of 1:1(semen/extenders). Then semen was filled into 0.5‐mL straws, and was frozen with vapour of liquid nitrogen at 4‐cm above surface of liquid nitrogen. After 3 min, straws were plunged in to liquid nitrogen. Thawing was performed at 40°C water baths for 15 s. Motility duration of the 10 mM HβCD treated spermatozoa at days 14 (228.98 ± 16.39) and 56 (199.66 ±21.78) were longer than other treatments. In day 56, the motility percentage in treatment with 10 mM was significantly higher (16.14 ± 2.54) (P < 0.05) compared with 5 mM treatment (8.75 ± 2.47) (P < 0.05). Therefore, it is recommended that 10 mM of HβCD can be used as an additive cryoprotectant for increasing cryopreserved spermatozoa quality in this species.  相似文献   

4.
This study examined the usage of a dry shipper for cryopreservation of Epinephelus septemfasciatus (Thunberg) spermatozoa. Milt was diluted 1:49 with 5% dimethyl sulfoxide plus 95% foetal bovine serum for cryopreservation. Computer‐assisted sperm analysis was used to analyse sperm motility, while fertilization and hatching trials were conducted to gauge the applicability of the cryopreservation method for aquaculture. We showed that cooling rates of the dry shipper were stable for 14 days and could be manipulated by the use of different sized freezing straws and use of a simple polystyrene foam container (5 × 5 × 12 cm and 1 cm thickness on all sides with the upper layer exposed). Dry shipper cryopreserved spermatozoa had significantly lower post‐thaw per cent motility and velocity than fresh sperm, but linearity of movement was unchanged. Fertilization and hatching rates were not significantly different at all tested sperm to egg ratios (3000:1–243000:1). The results indicated that 0.33 mL of milt when cryopreserved was sufficient to fertilize up to 450 g of oocytes. Application of this technology will help improve seed production in aquaculture and further develop breeding and genetics studies.  相似文献   

5.
The effects of extender composition and freezing rate on motility and fertility of frozen‐thawed Arctic char, Salvelinus alpinus, spermatozoa were investigated. Three freezing rates, two semen diluents and three cryoprotectants were tested. Semen frozen in 0.3 mol L?1 glucose diluent with 10% methanol as a cryoprotectant or in a diluent described by Lahnsteiner with 10%N,N‐dimethylacetamide (DMA) resulted in the highest sperm motility. Fertility was the highest for semen frozen in a glucose–methanol extender but was not significantly different than that for semen frozen in Lahnsteiner's diluent with 10% DMA. Dimethyl sulphoxide (DMSO) at 10% was a relatively ineffective cryoprotectant with either semen diluent. Semen frozen at 6 cm above the surface of liquid nitrogen resulted in a higher post‐thaw sperm motility and fertility than semen frozen at 5 cm. The addition of 7% fresh egg yolk to glucose diluent containing methanol or DMSO did not improve the fertility of frozen‐thawed spermatozoa. However, the addition of 7% fresh egg yolk to glucose–DMA extender significantly improved the fertilization percentages of frozen‐thawed spermatozoa. In conclusion, dilution of semen 1:3 in 0.3 mol L?1 glucose with 10% methanol and freezing 6 cm above the surface of liquid nitrogen (freezing rate of 40±8°C min?1, mean±SD from ?5 to ?55°C) is a promising protocol for cryopreservation of Arctic char semen.  相似文献   

6.
The effects of extender composition, cryoprotectant concentration and freezing and thawing on the fertilization efficiency of cryopreserved spermatozoa of Puntius gonionotus were evaluated. Computer‐aided motility analysis of semen was conducted to check the suitability of spermatozoa for cryopreservation after mixing with different extenders and cryoprotective agents (CPAs). Extender‐4 with an osmolality 260 mOsmol kg−1and pH 7.6 was used for the cryopreservation study. Among the CPAs, dimethyl sulphoxide (DMSO) was least toxic and more than 60% fertilization was achieved when used at 1.4 M at 0 °C for 10 and 30 min, whereas the toxicity of all CPAs to spermatozoa was evident when tested at 30 °C. Semen frozen at −16 °C min−1 with 1.4 M DMSO showed 70% fertilization, which was significantly higher (P<0.05) than other freezing rates. Samples thawed at 35 °C water showed a fertilization rate comparable with that of fresh semen. Computer‐assisted semen analysis of fresh and frozen semen after thawing showed variations in different types of motility in spermatozoa and in their class. There was no significant difference in motility before or after cryopreservation; however, significant differences could be observed in the average path velocity (VAP), straight line velocity (VSL) and curve linear velocity (VCL). Semen of silver barb could be cryopreserved with extender‐4 by addition of 1.4 M DMSO to a final cryopreservation medium (MED 2) cooled at a rate of −16 °C min−1, stored in liquid nitrogen (−196 °C) and utilized after thawing at 35±2 °C.  相似文献   

7.
For salmonid semen, the cryoprotective action of 10% methanol was compared with a 5% dimethyl sulphoxide (DMSO), 1% glycerol mixture, until now one of the most effective cryoprotectants. In Oncorhynchus mykiss (Walbaum), Salmo trutta L. f. fario, Salmo trutta L. f. lacustris and Salvelinus alpinus (L.), semen cryopreserved with both cryoprotectants yielded post-thaw fertilization rates of 90-100% of control with untreated semen at sperm-to-egg ratios of 1.8 × 106-2.4 × 106 spermatozoa per egg. However, at sperm-to-egg ratios of 0.9 × 106-1.2 × 106 spermatozoa per egg, semen cryopreserved with methanol had significantly higher fertilization rates than semen frozen with the DMSO/glycerol mixture. In other studies we obtained similar data for Coregonus sp., Salvelinus fontinalis (Mitchill), Thymallus thymallus (L.) and Hucho hucho (L.), proving that methanol is the most effective and generally applicable cryoprotectant for semen of the studied salmonid species. To facilitate the insemination of large egg batches we investigated the suitability of 1.2 ml and 5 ml straws for deep freezing of semen of Oncorhynchus mykiss, Salmo trutta f. fario, Salmo trutta f. lacustris and Salvelinus alpinus. With 1.2 ml straws the fertilization rates were similar to 0.5 ml straws when using lower freezing and higher thawing temperatures. The 5 ml straws resulted in a fertilization success of only about 40% of fresh semen control.  相似文献   

8.
This study aimed to determine the lowest effective concentration (LEC) of 2‐phenoxyethanol (2‐Phe) and essential oils (EOs) of Melaleuca alternifolia and Ocimum gratissimum able to induce anaesthesia in Astyanax bimaculatus. LEC was assumed to be the required concentration to carry out anaesthesia in less than 3 min with recovery in <5 min and sufficient to carry out routine aquaculture procedures. The following concentrations were tested: 2‐Phe (100, 200, 300, 400, 500 and 600 µl/L); M. alternifolia (200, 300, 400, 500 and 600 µl/L); and O. gratissimum (10, 20, 30, 40, 50, 60, 70, 100, 150, 250 and 300 µl/L). Results show that all three agents evaluated have effective anaesthetic properties for use in Astyanax bimaculatus with LEC for 2‐Phe, M. alternifolia and O. gratissimum of 200, 300 and 60 µl/L respectively.  相似文献   

9.
The effects of three extenders (Ginzburg fish ringer, Calcium‐free Hank's balanced salt solution, C‐F HBSS and sodium chloride, 0.9% NaCl) and four cryoprotectants (dimethyl sulphoxide, DMSO; dimethyl acetamide, DMA; methanol, MeOH and glycerol) in different concentrations (5%, 10% and 15%) on the motility, viability and fertilization rates of Mekong catfish (Pagasius bocourti) sperm were investigated. Sperm samples were transferred into 250‐μL French straws and sealed with a heated haemostat. The straws were then placed in a cryochamber. A computer‐controlled rate freezer (CL 3300) and programmable Cryogenesis, version 4 were used to regulate the freezing rate. The sperm samples were frozen at a rate of 10°C min?1 from 4 to ?80°C and then evaluated after 72 h. Of the three extenders used with each cryoprotectant, C‐F HBSS had the highest fertilization rate of 75% (93% of control). This was not significantly different from the control treatment (fresh sperm) when tested with DMSO as the cryoprotectant. The lowest fertilization rate of 27% (38% of control) was resulting from the combination of 15% glycerol and C‐F HBSS. This study found that fertilization, motility and viability rates in all of the experiments had a positive significant correlation (P < 0.001).  相似文献   

10.
Cryopreservation of Arctic charr Salvelinus alpinus (L.) semen was investigated using three diluents, three cryoprotectants [10% dimethyl sulphoxide (DMSO), 10% dimethylacetamide (DMA) or 20% glycerol] and three sizes of straw. The three diluents and three cryoprotectants were combined, resulting in nine extenders. One part semen was added to three parts extender, and motility was evaluated to assess the toxicity of six of the extenders. Semen in nine extenders was frozen in 0.5‐mL straws using liquid nitrogen vapour. Semen extended in 0.3 m glucose and each of the cryoprotectants was also frozen in 0.5‐mL, 1.7‐mL (flat) or 2.5‐mL straws. The freezing rate in each size of straw was measured. Fertility trials were conducted to determine the post‐thaw viability of the frozen semen. The motility of activated spermatozoa was higher in the DMA and DMSO extenders than in the glycerol extender. For the trial using 0.5‐mL straws, post‐thaw fertility results were higher for all extenders containing DMSO, or 0.3 m glucose and DMA, than for all other combinations of diluent and cryoprotectant. For the straw size comparison, the highest fertility was obtained for the 1.7‐mL straw using either DMSO or DMA and for the 2.5‐mL straw using DMSO. For all cryopreservation trials, fertility was low for extenders containing glycerol.  相似文献   

11.
Garlic (Allium sativum) has been shown to possess antimicrobial properties against a range of disease‐causing agents, including fish parasites. Our study aimed to investigate the potential use of garlic as a treatment against Cryptocaryon irritans infection, using guppies (Poecilia reticulata) as the fish model. Garlic was freeze‐dried, powdered, and used as it is or as an aqueous extract. The content of allicin, its main active ingredient, was 1.25 mg/g in garlic powder and 0.82 mg/ml in the aqueous extract, as determined by HPLC analysis. Aqueous garlic extract fully immobilized C. irritans theronts and protomonts within 20 and 40 min, respectively, following exposure to 5 µl/ml. Treatment trials were performed, in which infected fish were fed with garlic powder‐supplemented feeds (5%, 10%, or 20% supplementation), bathed in garlic aqueous extract (250 or 500 µl/L) and subjected to a combined treatment which included both feeding and bathing. Results revealed that the tested treatments failed to cure the infection, though reduction in infection intensity on the caudal fin, but not on the gills, was evident following dietary supplementation with 20% of powdered garlic in one of the trials.  相似文献   

12.
An experiment was carried to determine the plasma fenbendazole (FBZ) concentration and physio‐metabolic responses in juveniles of Labeo rohita (90 ± 4 g) after oral administration of single doses at 10, 20 and 50 mg, 20 mg FBZ/kg b.wt. in multiple times on 1st, 3rd and 7th day. The blood samples were collected at 0.5, 1, 2, 4, 8, 12, 24, 30, 48, 72, 96 and 120 hr, after single‐dose administration, and regularly (upto 15 day) in multiple dose. Plasma FBZ concentration was determined up to the limit of detection (LoD) of 0.09 µg/ml by HPLC. There was no parent drug detected in plasma for administration of 10 mg FBZ/kg b.wt. The drug attained the peak concentration (Cmax) 1.85 and 3.09 µg/ml in plasma at 4 hr (Tmax) after administration of 20 and 50mg FBZ/kg b.wt. respectively. Plasma FBZ was detectable up to 96 and 120 hr with concentration 0.09 ± 0.007 and 0.098 ± 0.006 µg/ml, respectively, after single‐dose administration of 20 and 50mg/kg b.wt. In case of multiple‐dose administration, the maximum concentration of FBZ was 1.01 ± 0.03 µg/ml on 7th day that was less than to the single dose at 50 mg/kg b.wt. However, FBZ was detected up to 11 day after multiple doses. The study revealed that the hepatic antioxidant enzymes activities like superoxide dismutase, catalase and glutathione‐S‐transferase were significantly affected by increasing FBZ in single and multiple doses. The results of the present study could reveal that single‐ or multiple‐oral administration of FBZ at 20 mg/kg b.wt. in feed as antihelminthic drug in L. rohita could be considered as the safe dose.  相似文献   

13.
The present experiment was aimed at studying the conditioning, maturation and natural spawning of orange‐spotted grouper, Epinephelus coioides, in a recirculatory aquaculture system (RAS). Thirty fish (n = 30; 3.35 ± 0.05 kg) were stocked in a circular tank of 125 m3 capacity fitted with an RAS for conditioning and broodstock development. After 15 days, 15 fish were implanted with 17 α methyl testosterone and letrozole at the rate of 5 mg and 0.2 mg/kg body weight, respectively, for conversion from female to male. The gonadal development started after 1 month, and by 90th day, 63.53 ± 3.78% and 2.07 ± 0.84% of the oocytes attained a size of 400–500 μm and 500–600 μm respectively. Natural spawning commenced in the RAS from 4th month onward after stocking and spawning continued round the year. The spawning pair showed courtship behaviour with a typical vertical burst of swimming just before release of gametes. The total number of eggs spawned during 1 year was 47.23 million with spawning frequency varying form 5 to 13 times per month. The association of spawning events with new moon day (lunar cycle) weakened as time progressed. The mean monthly fertilization and hatching rates varied from 77.80 ± 3.34% to 83.70 ± 1.76% and 82.80 ± 4.21% to 88.33 ± 1.39% respectively. The study proved that RAS is an efficient system that provides a stable, controllable and conducive environment for year‐round natural breeding of orange‐spotted grouper.  相似文献   

14.
In this study, total phenolic and flavonoid contents of grapefruit peel extract (GPE) were equal to 117.3 ± 0.3 µg of gallic acid/mg and 39.30 ± 0.1 µg of quercetin/mg respectively. Caspian white fish (n = 180, 4 ± 0.9 g body weight) was fed with supplemented diets, including 0, 6.25, 12.5 and 25 mg of GPE/kg for 60 days at 25 ± 1°C. The growth performance was markedly improved in fish fed with 25 mg/kg of GPE compared to others (p < 0.05). Moreover, fish fed with 25 mg/kg of GPE showed a significant increase in red blood cell (2.65·106 cell/mm3), white blood cell (17.75·103 cell/mm3), packed cell volume (48%) and haemoglobin concentrations (8.75 g/dl) compared to the control (p < 0.05). However, the highest alanine aminotransferase (140 U/L), alanine transaminase (14.5 U/L), and alkaline phosphatase (18.5 U/L) were observed in control group. Morphological analysis of intestine revealed the highest amount of villus width (8.4 µM), height (32.86 µM) and surface area (342.7 µM2) in fish fed with 25 mg/kg of GPE (p < 0.05). In conclusion, supplementing feed with GPE at 25 g/kg can improve growth performance and haemato‐biochemical parameters of Caspian white fish fry.  相似文献   

15.
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.  相似文献   

16.
Effects of dietary supplementation of Tribulus terrestris (TT) and 17α‐methyl testosterone (MT) on growth performance and reproductive efficiency of male Nile tilapia, Oreochromis niloticus (N = 75, initial weight 60 ± 3.251 g), were evaluated. Five experimental treatments in triplicate were fed on basal diet (30% crude protein) (control group, CG), basal diet containing 60 mg MT (MT group) and basal diet containing 250, 500 and 750 mg/kg diet TT (TT250, TT500 and TT750 groups), respectively, for consecutive 45 days. Results revealed that the fish received TT in diet showed better growth performance (final body weight, weight gain, feed conversion ratio, specific growth rate, testes weight and gonadosomatic index) compared with MT and CG (p < .05), as well as for haemoglobin content and red blood cell count (p < .05). The highest level of testosterone, 11‐keto testosterone, sperm concentration and vitality was recorded in TT750 followed by TT500, TT250 and MT groups, while the lowest value was recorded in the CG (p < .05). The highest seminiferous tubule diameter was recorded in TT750 followed by TT500, TT250 and CG, while the lowest diameter was recorded in the MT group (p < .05). Fish received TT showed normal dilated seminiferous tubule filled with large amount of free sperms within their lumen in a dose related effect. In conclusion, male Nile tilapia dietary supplementation with different inclusions of Tribulus terrestris extract specially 500 and 750 mg/kg diet improves growth performance, health condition, semen quality and reproductive efficiency without any harmful effect on water quality.  相似文献   

17.
The summer flounder, Paralichthys dentatus L., is a high‐value species and considerable research has been conducted to determine practices conducive for its culture. As milt can be limited in this species, experiments were conducted to develop a practical sperm cryopreservation protocol for hatchery use. Two dilution ratios (1:2 and 1:4; sperm:extender), 2 diluents (saline and sucrose‐based), 2 cryoprotectants (10% DMSO and 12% glycerol) and 3 freezing rates (?5, ?10 and ?15°C min?1) were evaluated using differential staining to assess post‐thaw sperm survival. Seven combinations of the factors examined reduced post‐thaw viability by less than 30%. The average viability of sperm from fresh, pooled flounder milt (67.2 ± 2.9%) was not different from that of thawed milt diluted 1:4 with sucrose diluent (10% DMSO) frozen at ?5°C min?1 (38.4 ± 7.7%) and fertilization and hatch success were not different in trials using fresh or thawed, cryopreserved sperm. From these experiments a practical sperm cryopreservation method was developed, but further refinement of the freezing protocol is necessary to optimize results.  相似文献   

18.
With an overarching objective of improving the hatchery production of Atlantic salmon (Salmo salar) all‐female progeny, this study comparatively evaluated the reproductive parameters between normal (genotype XY) and neo‐males (genotype XX). Four normal (XY) and seven neo‐ (XX) males, from the same brood stock, distinguished by their ability to or lack of expressing semen, respectively, were comparatively evaluated. The left testicular lobe was used for histomorphometric analyses, while the right for semen collection and sperm quality analyses. Histomorphometric observations revealed that neo‐male testes are irregularly shaped, and have poorly formed seminiferous ducts, higher proportions of interstitial tissue and lower gonadosomatic index (p < .05). In addition, hypertrophied and cyst forming Sertoli cells were found in these individuals which collectively appear to form a physical barrier, precluding the semen collection by standard stripping techniques and reducing sperm quality. Particularly, semen motility (80.69 ± 2.4% and 57.2 ± 36.5% for XY and XX respectively) and duration of motility (99.31 ± 28.03 s and 66.84 ± 23.83 s for XY and XX respectively) of neo‐males were most compromised (p < .05). Interestingly, the TUNEL assay indicated no signs of apoptotic tissue suggesting that the histological differences may relate to delayed physiological/sexual maturity of neo‐males.  相似文献   

19.
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.  相似文献   

20.
The characterization of sperm motility patterns, particularly post‐activation changes, is the first step in setting up species‐specific protocols involving gamete management and embryo production, for both aquaculture and laboratory research purposes. This study is aimed at the characterization of the sperm motility pattern of the purple sea urchin Paracentrotus lividus. Semen samples were individually diluted in artificial sea water for sperm motility activation. They were then incubated at 18°C for up to 24 hr. Motility was evaluated on dilution, and 1 hr, 3 hr and 24 hr after activation, by computerized analyser. The semen fertilization capacity was also evaluated. Under our experimental conditions (dilution 1:1,000 in artificial sea water plus 0.05% BSA, 18°C, in the dark), P. lividus semen remained viable for up to 24 hr, as the total motile sperm and the fertilization percentages did not change significantly during the incubation time. In contrast, the mean curvilinear velocity and the subpopulation of rapid sperm (those having a curvilinear velocity > 100 µm/s) slightly but significantly decreased after 3 hr, thereafter remaining unchanged for up to 24 hr after activation. In conclusion, our results show that diluted P. lividus semen can be used for a longer period than that of most fish species, with no need for motility inhibition procedures, supporting its wider use in laboratory research. In addition, the development of artificial fertilization protocols for aquaculture production is simplified by long‐lasting sperm motility.  相似文献   

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