盐类对南方根结线虫迁移能力和侵入能力的影响

为探讨无机盐和有机盐对南方根结线虫的作用方式,在室内利用沙柱法和染色法测定了二者对南方根结线虫2龄幼虫迁移能力及侵入番茄根部能力的影响。结果表明,随着各盐处理浓度的增加,南方根结线虫2龄幼虫的迁移能力和侵入能力显著降低。在4μg/m L KSCN、C6H8O7、KCl、KH2PO4、KHCO3和Mn SO4·H2O沙柱中,15 d后2龄幼虫的平均迁移距离最小,分别为0、0.33、0.52、0.53、0.55和0.58 cm,而对照为0.94 cm。2龄幼虫在Ba NO3、K2HPO4、KHCO3、Cs Cl和KSCN各浓度沙柱中的平均迁移距离均小于0.94 cm。2μg/m L NH4NO3、KH2PO4、(NH4)2CO3、Cs Cl、(NH4)2SO4、Na SCN和NH4SCN处理15 d后,2龄幼虫对番茄根部的侵入率最低,分别为1%、2.33%、1.33%、2%、2%、1%和1.67%。表明对2龄幼虫侵入番茄根部抑制效果最明显的盐为NH4NO3、KH2PO4、(NH4)2CO3、Cs Cl、(NH4)2SO4、Na SCN和NH4SCN。     

Single layer centrifugation of stallion spermatozoa consistently selects the most robust spermatozoa from the rest of the ejaculate in a large sample size

Reasons for performing study: An improvement in sperm quality after single layer centrifugation (SLC) has been seen in previous studies using small sample sizes (for example, n = 10 stallions). There is a need to investigate whether this improvement is repeatable over several breeding seasons with a larger number of stallions (n ≥ 30 stallions). Objective: To make a retrospective analysis of the results of SLC performed on more than 250 sperm samples (176 ejaculates) from 31 stallions in 3 consecutive breeding seasons. Methods: Sperm quality (motility, proportion of morphologically normal spermatozoa and the proportion of spermatozoa with undamaged chromatin) was assessed before and after SLC. Results: All parameters of sperm quality examined were significantly better in sperm samples after SLC than in their unselected counterparts (P<0.001 for each parameter). The yield of spermatozoa obtained after SLC was influenced by the type of extender used and also by the concentration of spermatozoa in the original ejaculate, with fewer spermatozoa being recovered when the loading dose contained a high concentration of spermatozoa. The optimal concentration was approximately 100 × 10⁶/ml. Sperm concentration in the samples loaded on to the colloid influenced the sperm yield while the type of semen extender affected sperm quality and survival. Furthermore, the scaled‐up SLC method was found to be suitable for use with a range of ejaculates, with similar sperm kinematics being observed for standard and scaled‐up preparations. Conclusions: SLC consistently improved the quality of stallion sperm samples from a large number of ejaculates. The method could be scaled‐up, allowing larger volumes of ejaculate to be processed easily from a wide range of stallions.     

Production of monoclonal antibody against recombinant bovine sex-determining region Y (SRY) and their preferential binding to Y chromosome-bearing sperm

Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms.     

离心除精浆对提高冷冻精液品质的作用

采用离心洗涤法去除精浆后,对精液进行冷冻处理。发现:1、除精浆精子冻后的活率、Ⅰ类顶体率和精子畸形率与未除精浆组存在显著差异(P<0．05);2、除精浆使稀释后精液精浆中GOT和LDH活性明显升高(P<0．05)。从平衡后到解冻后各组GOT和LDH活性都呈上升趋势。但除精浆试验组比未除精浆试验组上升缓慢;3、除精浆试验组精子的穿卵率、卵内平均精子数都显著高于未除精浆试验组(P<0．05);4除精浆离心洗涤液用生理盐水和脱脂乳-卵柠液．两者之间无显著差异(P>0．05)。     

Seminal plasma composition and its relationship with physical spermatological parameters of Grass carp (Ctenopharyngodon idella) semen: with emphasis on sperm motility

In this research, the mineral and organic composition of the seminal plasma, physical spermatological parameters and their physiological relationships were investigated in grass carp (Ctenopharyngodon idella). The seminal plasma contained 98.14±5.23 mM L^?1 (Na⁺), 380.85±25.95 mM L^?1 (K⁺), 30.25±4.96 mg dL^?1 (Ca²⁺), 19.16±1.70 mEq L^?1 (Mg²⁺), 1.36±0.11 mg dL^?1 glucose, 0.37±0.08 g dL^?1 total protein, 12.02±1.18 mg dL^?1 cholesterol, 14.85±1.50 mg dL^?1 triglyceride and 43.5±9.56 mg dL^?1 urea. The following spermatological parameters were found: sperm volume 14.44±1.16 mL, sperm motility 80.60±1.55%, movement duration 67.68±4.32 s, density 15.43±0.72 × 10⁹ mL^?1, total density 337.43+45.86 × 10⁹ and pH 7.24±0.17. The Na⁺ and Ca²⁺ ions correlated negatively with spermatozoa motility (r=?0.453, P>0.05 and r=?0.192, P>0.05) respectively. The K⁺ ion correlated positively with spermatozoa motility (r=0.545, P>0.05). But a statistically significant correlation was not observed between sperm motility and seminal plasma parameters. The following correlations were observed between mineral and organic components. The Mg²⁺ was positively correlated with glucose and cholesterol (r=0.692, P<0.05 and r=0.680, P<0.05) respectively. A highly significant positive relationship was also found between Mg²⁺ and total protein (r=0.837, P<0.01). On the other hand, a significantly negative relationship was found between Ca²⁺ and triglyceride (r=?0.639, P<0.05). These parameters should be considered when developing procedures for either artificial fertilization or for cryopreservation of grass carp sperm.     

Cryopreservation effects on the sperm quality of cachama blanca Piaractus brachypomus (Cuvier 1818)

The effects of straws volume, cryoprotectants and thawing temperatures were evaluated on the sperm quality of cachama blanca Piaractus brachypomus (Cuvier), an important Colombian fish species. Sexually mature fish were induced to ovulation or spermiation with a carp pituitary extract. A pool of suitable sperm samples was diluted in glucose, egg yolk, dimethyl sulphoxide (DMSO‐10%), methanol (MET‐10%) or ethylene glycol (ETG‐5%) and packed in 0.5, 2.5 or 5.0 mL straws and frozen in nitrogen vapour. The thawing process was performed in a 35 or an 80 °C water bath. The fertility was evaluated after 6 h post fertilization. The highest motility percentage (33 ± 3%) was observed with sperm cryopreserved with DMSO, packed in 5 mL straws and thawed at 35 °C. The treatments with DMSO and MET packed in 0.5 and 5.0 mL straws and thawed at 35 °C showed the highest fertility (higher than 71%) and the lowest fertility was obtained with MET‐2.5 mL (9 ± 5%). In all the treatments, a significant decrease in the sperm quality was observed at 80 °C. Sperm cryopreserved with DMSO‐10% or MET‐10%, packed in 2.5 or 5.0 mL straws are suitable to achieve acceptable fertilization and to fertilize high amounts of eggs.     

Cryopreservation of strip spawned sperm using non‐programmable freezing technique in the blue mussel Mytilus galloprovincialis

In this study, a non‐programmable freezing technique has been developed with the strip spawned blue mussel (Mytilus galloprovincialis) sperm. The key parameters optimized including (1) cryoprotectant agents (CPAs); (2) cooling temperature; (3) thawing temperature; (4) sugar and amino acid supplementation and (5) sperm to egg ratio. The fertilization rate and/or integrity of plasma membrane and acrosome were used as sperm quality assessment indicators. The highest post‐thaw sperm fertilization rate of 95% was achieved, when sperm were cryopreservated in 8% dimethyl sulfoxide at 7.8 cm above the liquid nitrogen surface and thawed in a 60°C seawater bath. The addition of glucose, sucrose or trehalose in dimethyl sulfoxide did not, whereas 0.8% glycine did significantly improve the post‐thaw sperm fertilization rates. The fluorescent evaluation has demonstrated that the addition of glycine significantly improved the post‐thaw sperm acrosome integrity, revealing a positive role of glycine in the improvement of post‐thaw sperm quality in blue mussels.     

虾夷扇贝精子的主要生理特性与低温保存

试验结果表明,虾夷扇贝精子的最适盐度为23.2～32,最适pH 7.0～8.5.4℃,未稀释精液样品保存5 d后,精子仍有(26±4)%的活力,而经海水稀释后的精液保存70 h后,最高活力仅为(11±1.41)%,用抗冻液稀释的精液,4℃保存5 h后,得到最高的精子活力仅为(24.7±4.16)%.添加抗冻剂后,-18℃的保存效果明显低于4℃保存的未稀释样品.用4℃保存的未稀释精液与栉孔扇贝卵子进行杂交试验,保存3 d内的精子获得的授精率与对照组差异不显著(P0.05).杂交组的幼虫可以正常发育至D形幼虫,与自交组无区别.