Artificial induction of maturation and fertilization in the Japanese eel, Anguilla japonica   总被引：4，自引：0，他引：4  

H. Ohta  H. Kagawa  H. Tanaka  K. Okuzawa  N. Iinuma  K. Hirose 《Fish physiology and biochemistry》1997,17(1-6):163-169

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.  相似文献   

Ionic composition and osmolality of paddlefish (Polyodon spathula, Acipenseriformes) seminal fluid     

Otomar Linhart  Steve D. Mims  Boris Gomelsky  Ana E. Hiott  William L. Shelton  Jacky Cosson  Marek Rodina  David Gela  Jan Bastl 《Aquaculture International》2003,11(4):357-368

Changes in ionic composition as Na⁺,K⁺, Ca²⁺ and Mg²⁺, osmolality inseminal fluid, percentage of motile spermatozoaand velocity were investigated in response toCPP and different dosage of LHRHa. The lowestvelocity of sperm was observed after use CPPtreatment. The velocity of spermatozoa,significant main effect of the treatment(P < 0.0001) and the time of sperm collection(P < 0.0104) were evaluated. The osmolality ofseminal fluid was different betweenexperimental groups of LHRHa (48.0–62.7mOsmol.kg^–1) and CPP (33.0–46.3mOsmol.kg^–1) treatments. The osmolalitywas significantly higher on the first day andone-half, then declined on day three, rangingfrom 33.0 to 62.7 mOsmol.kg^–1. Analysisof variance showed significant main effects ofthe treatment (P < 0.0001) and the time ofsperm collection (P < 0.0002) on the osmolalityof seminal fluid. The level of Na⁺ andK⁺ ion was different between experimentalgroups of LHRHa and CPP treatment. The highestconcentration of 11.11 mmol.l^–1 wasobserved at Na⁺ ion. Then theconcentrations declined on the level 1.56, 0.52and 0.36 mmol.l^–1 for K⁺, Ca²⁺and Mg²⁺ ions, respectively. There werehighly positive correlations between osmolalityof seminal fluid and dosage of LHRHa treatment(r = 0.84), velocity of spermatozoa andosmolality of seminal fluid (r = 0.57) andosmolality of seminal fluid and Na⁺concentration at seminal fluid (r = 0.70).Injection with LHRHa increased quality of spermas velocity of sperm, level of Na⁺,K⁺ and osmolality at seminal fluidcompared to CPP treatments.  相似文献   

Long-term cryopreservation of spermatophore of the giant freshwater prawn, Macrobrachium rosenbergii (de Man)     

Khattiya Akarasanon  Praneet Damrongphol  & Wandee Poolsanguan 《Aquaculture Research》2004,35(15):1415-1420

Long‐term cryopreservation of the giant freshwater prawn, Macrobrachium rosenbergii, spermatophores using glycerol (Gly) and ethylene glycol (EG) as cryoprotective agents (CPAs) was studied. The tolerance of sperm to cryopreservation was evaluated on the basis of sperm survival and fertilizing ability. The survival of the sperm was determined by trypan blue staining, while the fertilizing ability was assessed from artificial insemination of the cryopreserved spermatophores. The rates of embryo survival on day 5 after spawning and of spermatophores capable of producing embryos survived to hatching were determined. Storage of spermatophores at ?20°C without CPA for a short period of up of 1–5 days decreased the sperm survival significantly and did not preserve fertilizing ability. Preservation at ?20°C in the presence of 10% or 20% Gly or of 10% or 20% EG offered a simple and efficient short‐term storage up to 10 days. For a long‐term storage, cryopreservation in the presence of 20% EG at ?196°C was more efficient than at ?20°C. High sperm survival rates and high fertilizing ability were recorded from those cryopreserved at ?196°C for up to 150 days. High sperm survival rates with moderate levels of fertilizing ability were obtained from those cryopreserved at ?20°C for not more than 30 days. The results indicate that preservation at ?196°C with 20% EG is a suitable procedure for long‐term storage of the giant freshwater prawn spermatophores.  相似文献   

Role of sodium bicarbonate on the initiation of sperm motility in the Japanese eel   总被引：1，自引：0，他引：1  

Satoru  TANAKA  Tomoko  UTOH  Yoshiaki  YAMADA  Noriyuki  HORIE  Akihiro  OKAMURA  Atsushi  AKAZAWA  Naomi  MIKAWA  Hideo P  OKA  Hisashi  KUROKURA 《Fisheries Science》2004,70(5):780-787

ABSTRACT:   In order to find out the role of sodium bicarbonate (NaHCO₃) on the initiation of sperm motility in the Japanese eel Anguilla japonica , interactions were investigated between NaHCO₃ and various reagents (K⁺ channel blocker 4-aminopyridine [4-AP], ammonium chloride [NH₄Cl], sodium acetate and calcium chloride [CaCl₂]) that could regulate internal factors (intracellular K⁺, intracellular pH [[pH]_i] and intracellular Ca²⁺) in sperm motility. Contradictory effects of NaHCO₃ were observed (i.e. an inhibitory effect when 4-AP was absent and a promoting effect when 4-AP was present). Sodium bicarbonate inhibited the initiation of sperm motility in the Japanese eel. However, NaHCO₃ restored the motility of immotile sperm that 4-AP inhibited. The inhibitory effect of NaHCO₃ disappeared with the addition of NH₄Cl, which raised [pH]_i, but the promoting effect was not affected by [pH]_i. Although NaHCO₃ recovered motility in the presence of 4-AP, this recovery was also observed with the addition of CaCl₂ instead of NaHCO₃. In the initiation of sperm motility in the Japanese eel, two roles for NaHCO₃ are suggested: an inhibitory role relating to the regulation of [pH]_i and a promoting role relating to the uptake of another initiation factor, which could be Ca²⁺.  相似文献   

Short-term storage and cryopreservation of Urechis unicinctus (Echiura: Urechidae) sperm   总被引：2，自引：0，他引：2  

Kyoung Ho Kang  Ming Yu Shao  Kang Hee Kho  & Zhi Feng Zhang 《Aquaculture Research》2004,35(13):1195-1201

In the present study, attempts were made to preserve Urechis unicinctus sperm at 4°C. Cryopreservation procedures were optimized for various cryoprotectants and freezing rates, equilibration times and dilution ratios. During short‐term storage, the motility of undiluted sperm was extended for 6 days of cold storage,and in 70% and 100% artificial seawater only persisted for 2 and 4 days respectively. The survival rate of undiluted sperm was maintained at a high level accordingly. After cryopreservation, the highest motility and survival rate (41.5±2.2%) were obtained in 15% dimethyl sulphoxide (Me₂SO) using a freezing rate of 30°C min^?1. After thawing the sperm cryopreserved in glycerol lost almost all motility. The motility and survival rate of post‐thawing sperm did not show significant differences after 8 and 15 min equilibration using 15% Me₂SO as cryoprotectant; the values were significantly higher than those of 2 min equilibration. Comparisons of motility and survival rate between treatments pooled by dilution ratio showed that the effect of 1:1 ratio (sperm volume to cryoprotectant volume) was best. There was no difference between 1:3 and 1:5, and other ratioswere significantly worse.  相似文献   

牛精液冷冻保护剂研究进展     

贾永宏  朱庆超  杨海涛  武致存  高璞  李梅  胡建宏 《家畜生态》2014,(1):5-8

牛的人工授精技术是目前应用最为广泛的动物繁殖技术之一,对牛的遗传改良做出了巨大贡献。但是,在冷冻一解冻过程中仍然有大约40％～50％的精子失去活性,限制良种公牛种用性能的发挥。论文在分析精子冷冻保存原理的基础上,阐述了渗透性、非渗透性冷冻保护剂以及低密度脂蛋白对牛精子的冷冻保护作用,以期为开发新型冷冻保护剂的研究提供一定参考,进一步提升牛冷冻精液的质量。  相似文献   

Addition of oxytocin to semen extender improves both sperm transport to the oviduct and conception rates in pigs following AI     

Tukasa Tinen  Teiichi Akiyoshi  Manabu Mori  Hisanori Teshima 《Animal Science Journal》2014,85(1):8-14

Oxytocin (OXT) contained in boar semen is known to produce uterine contraction; therefore, we hypothesized that the co‐injection of OXT with sperm would improve artificial insemination (AI) using liquid or frozen‐thawed boar sperm. We initially examined whether OXT added to semen extender improved sperm transport to the oviduct. Although the addition of OXT did not affect the fresh or frozen‐thawed sperm motility or acrosomal integrity, it significantly increased the number of sperm in the oviduct at 6 h after AI injection with OXT, as compared with the control (P < 0.05). Moreover, some sperm were observed in the sperm reservoir of the isthmus in the OXT treatment group, whereas few sperm were observed in the control. When OXT was added to the semen extender immediately prior to AI, the conception rates were significantly higher in both fresh semen and frozen‐thawed semen than in the control group (P < 0.05: liquid, 87.5% vs. 70.5%; frozen‐thawed, 89.8% vs. 75.0%). From these results, we concluded that the addition of OXT to the semen extender assisted in sperm transportation from the uterus to the oviduct, which resulted in improved reproductive performance.  相似文献   

Acquisition of the potential for sperm motility in steelhead (Oncorhynchus mykiss): effect of pH on dynein ATPase     

J. Woolsey  R.L. Ingermann 《Fish physiology and biochemistry》2003,29(1):47-56

Salmonid sperm pre-incubated at extracellular pH (pH_e) values less than about 7.4 do not become motile upon water activation whereas sperm maintained above about pH 8.0 demonstrate maximal motility upon activation. The basis for this permissive effect of elevated pH_e on sperm motility is not known. Since it is conceivable that the pH sensitivity of dyneinATPase (the molecular motor that drives flagellar movement) could be the basis of, or contribute to this pH dependency, the pH sensitivity of this enzymatic activity was evaluated in membrane-permeabilized axonemes (isolated flagella) ofsteelhead sperm. DyneinATPase activity was found to be sensitive to pH. This activity in permeabilized axonemes was about 3.5-fold higher at pH 7.6 compared to 7.0. To determine whether the pH sensitivity ofATP regeneration might affect the interpretation of the effect of pH on dyneinATPase activity, the pH sensitivities of creatine kinase and adenylate kinase were established. The rates ofATP generation by these enzymes were insensitive to pH between 6.5 and 8.0. The results of these studies are consistent with the hypothesis that prior maintenance at pH_e, in part, controls the potential for sperm motility upon water activation via an influence on dyneinATPase activity. However, the potential for motility ofsteelhead sperm is particularly sensitive to prior maintenance at pH_e values between about 7.4 and 8.0 whereas the dyneinATPase activity of permeabilized axonemes was particularly sensitive to pH values between 7.0 and 7.6. Phosphorus NMR spectroscopy was used to determine that sperm intracellular pH (pH_i) increased with increasing pH_e between 7.0 and 8.5 and pH_i was, on average 0.4–0.5 pH units lower than pH_e. Therefore the pH_e sensitivity of the potential for motility appears to correspond to the pH_i sensitivity of dyneinATPase activity. The data indicate that pH_i is directly related to pH_e and that prior incubation at pH_e may, in part, control the sperm's potential for motility upon water activation via an influence on dyneinATPase activity.  相似文献   

Seasonal changes in sperm production and quality in the red porgy Pagrus pagrus (L.)     

Constantinos C Mylonas  Maria Papadaki  & Pascal Divanach 《Aquaculture Research》2003,34(13):1161-1170

Cultured red porgy Pagrus pagrus (L.) males (n=6) were sampled every 2 weeks for milt, in order to monitor changes in sperm quality parameters during a whole spawning period. On 11 January 2001, 60% of the fish were spermiating, increasing to 100% in mid‐February and dropping to 30% by mid‐April. Sperm density showed a slight increasing trend, with mean values ranging between 8.6 and 23.7×10⁹ spermatozoa mL^?1. Sperm motility percentage exhibited a significant improvement during the spawning season (analysis of variance (anova ) P=0.0001). The duration of forward motility for the major part of the monitoring period ranged between 2 and 4 min. Red porgy spermatozoa maintained their viability for many days after whole storage of milt at 4°C. During the monitoring period there were significant changes in the mean duration of sperm survival after cold storage, ranging from 5 to 12 days. The total volume of expressible milt was maximal on 28 March, increasing from a mean value of 1.7 mL to 5.3 mL kg^?1. Milt production of captive‐reared red porgy does not appear to be limiting, when compared with the volume of expressible milt produced by other cultured marine fishes.  相似文献   

Motility Parameters of Rainbow Trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) Spermatozoa in Relation to Sequential Collection of Milt,Time of <Emphasis Type="Italic">Post-mortem</Emphasis> Storage and Anesthesia     

G.?J.?Dietrich  R.?Kowalski  M.?Wojtczak  S.?Dobosz  K.?Goryczko  A.?CiereszkoEmail author 《Fish physiology and biochemistry》2005,31(1):1-9

The present study investigated the effects of sequential collection of milt, time of post-mortem storage and anesthesia on rainbow trout (Oncorhynchus mykiss) sperm motility parameters (using computer-assisted sperm analysis – CASA) as well as seminal plasma osmolality and sperm concentration. The post-mortem storage and time of anesthesia altered motility characteristics of rainbow trout sperm to different extents. The moderate impact of time of anesthesia was manifested in a shortened duration of sperm motility after 10 min exposure of fish to anesthetic. The prolonged post-mortem storage (≥40–60 min), in addition to lowering sperm motility duration, also significantly influenced sperm motility parameters, such as sperm velocities, percentage of motile sperm and sperm trajectory parameters. These results clearly demonstrate that when milt from sacrificed fish is used for sperm motility studies, the time of post-mortem storage significantly alters sperm motility characteristics. Since sperm motility rate and swimming velocity could predict fertilizing ability, detrimental effects of prolonged post-mortem storage may lead to reduced fertilization success. Sperm concentration and seminal plasma osmolality were lower in the first fractions and increased with successive collections of milt. It suggests the presence of urine contamination of the first milt fractions which were collected by stripping. Therefore, testing of sperm concentration and/or seminal plasma osmolality should be mandatory while handling stored milt.  相似文献