Relationship of cysteine‐rich secretory protein‐3 gene and protein with semen quality in stallions

Cysteine‐rich secretory protein‐3 (CRISP‐3) and some of its nonsynonymous polymorphism have been related to the fertility and freezability of stallion semen; however, the role of the CRISP‐3 gene and its seminal plasma protein in the raw semen quality is still unknown. The aim of this study was to evaluate the relationship of CRISP‐3 with semen quality in stallions. DNA was obtained from blood samples of 100 stallions, from which 30 stallions were randomly selected to obtain 60 ejaculates. Through PCR amplification and sequencing, the variation of four nonsynonymous SNPs from CRISP‐3 was identified and haplotypes were derived. Semen quality was assessed through the total motility (MOT), sperm vitality (SV), normal morphology (NM), functional integrity of membrane (MI) and a seminal quality index (SQi). CRISP‐3 protein content of seminal plasma (SP) was determined by ELISA. The effect of the genotype, the haplotype and the concentration of the CRISP‐3 protein on the seminal quality were evaluated through generalized linear models and linear regression analyses. Homozygous genotypes for SNP1, SNP2 and SNP3 and the heterozygous genotype for SNP4 showed a positive effect on seminal quality. Different haplotypes with positive effect on MOT, SV, NM, MI and SQi were identified. The allelic substitution analysis resulted in positive regression coefficients for MOT (SNP2) and MI (SNP2 and SNP3). A high level of CRISP‐3 resulted in a higher MOT and SQi. It is concluded that the quality of stallion semen is influenced by the genotype of CRISP‐3 and the concentration of CRISP‐3 protein in SP.     

Modulation of seminal plasma content in extended semen improves the quality attributes of ram spermatozoa following liquid preservation at 3–5°C

Seminal plasma (SP) is known to induce motility and capacitation in spermatozoa curtailing their lifespan when preserved. Hence, this study was conducted to examine the effects of removal of SP from sperm surface prior to liquid preservation either by high dilution (1/15) or by washing and the poststorage treatment with SP (15% and 25%, v/v) on the quality attributes of liquid‐preserved ram semen. Over the period of storage, the rapid motility (66.0% and 71.1% vs. 58.3%), straightness (87.1% and 82.1% vs. 79.4%), average path velocity (152.3 and 152.0 µm/s vs. 133.3 µm/s) and the straight‐line velocity (131.3 and 127.8 µm/s vs. 108.5 µm/s) were significantly (p < 0.05) higher in both the high‐dilution and wash groups as compared to the control (1/3 dilution). The functional membrane integrity (82.3% vs. 77.2%) and noncapacitated sperm count (65.0% vs. 58.7%) were also significantly (p < 0.05) higher in the high‐dilution and wash groups, respectively, as compared to the control. The poststorage treatment of sperm with SP significantly (p < 0.05) increased the functional membrane integrity (70.1% vs. 53.8%) and most of the motility attributes as compared to the control (without SP). In conclusion, both the removal of SP prior to liquid preservation and poststorage treatment with SP significantly improved the quality attributes of ram spermatozoa.     

山羊除精浆冷冻精液的研究进展

山羊精浆中含有卵黄凝结酶等物质 ,对冷冻精液品质造成不良影响 ,应用离心处理等方法除去精浆后 ,可明显提高解冻后精子的品质。文章综述了这一领域的研究近况 ,并对其予以展望。     

ATP content and sperm motility of extended bovine semen under different storage conditions.

The effects of incubation temperature (+20 degrees C vs +35 degrees C) and media type on the ATP content and motility of spermatozoa were determined in fresh bovine semen in order to develop a method for assaying post-thaw quality. Semen was obtained from 3 bulls at 2 occasions. The spermatozoa were washed using a Ficoll-containing medium before being resuspended in each of 4 different media (I. 0.9% NaCl; II. Tris-buffer solution; III. seminal plasma; IV. seminal plasma+Tris-buffer solution) and incubated for 6 h. The least-squares means for ATP content were higher (p less than or equal to 0.05) at +20 degrees C than +35 degrees C for all media except no. I. By contrast, the least-squares means for sperm motility were higher (p less than or equal to 0.05) at +35 degrees C than at +20 degrees C in media II and III. A decrease over time in ATP content and motility at both temperatures was also observed. The single most important factor responsible for changes in ATP content and sperm motility was the temperature and the medium, respectively.     

Biochemical Composition of Seminal Plasma and Annual Variations in Semen Characteristics of Jundiá <Emphasis Type="Italic">Rhamdia quelen</Emphasis> (Quoy and Gaimard,Pimelodidae)

This study investigated the composition of milt of the South American silver catfish (Rhamdia quelen) or jundiá. The semen was taken from jundiá in different periods during the four seasons. The biochemical composition of seminal fluid and the characteristics of sperm were analyzed. The semen quantity which can be extracted per fish in one day was 0.95 ± 0.08 ml during spring (maximum) and 0.24 ± 0.03 ml during winter (minimum). Sperm density (spermatocrit) showed higher values in the spring (75.1 ± 1.3%) decreasing slightly afterwards and reaching 63.0 ± 2.4% to 65.0 ± 2.2% in the fall and winter. Immediately after water dilution, 90–100% of the spermatozoa presented vigorous straightforward motility that remained for at least 20 s. The total duration of the motility was 47.9 ± 1.3 s in the spring and 38.6 ± 0.6 s in the other seasons (P < 0.05). This pattern of motility is maintained for more than 2 h after storage of the milt at room temperature. The pH from 5 to 10 of the water dilution does not influence the sperm motility. The mean seminal pH and osmolality values were 8.7 ± 0.07 and 274.8 ± 11.2 (mOsm/kg), respectively. The ion concentration was: Na 153.7 ± 2.4, K 10.7 ± 2.4, Cl 139.4 ± 2.1, Ca 4.2 ± 0.2, Mg 0.9 ± 0.05, P 0.9 ± 0.08 (mEq/l). The total protein was 0.6 ± 0.05 mg/dl and cholesterol concentration was 13.9 ± 0.9 mg/dl.     

The presence of uric acid, an antioxidantive substance, in fish seminal plasma

High concentrations of uric acid in seminal plasma of a range of teleost fish species are reported for the first time. Concentrations of urate amounted to 223.4; 121.9–130.0, 355.9, 735.6, 124.0, 192.7 and 148.0 M for rainbow trout (Oncorhynchus mykiss), yellow perch (Perca flavescens), muskellunge (Esox masquinongy), Northern pike (Esox lucius), carp koi (Cyprinus carpio), bream (Abramis brama), and tench (Tinca Tinca), respectively. Uric acid is an important anti-oxidant, therefore, it has a potential role in protection of fish spermatozoa against oxidative damage.     

Chemical composition and osmolality of seminal fluid of Acipenser persicus; their physiological relationship with sperm motility

Determination of semen quality is necessary to understand the basic biochemical processes occurring during motility of sperm and during fertilization to evaluate the reproductive ability of different fish species and to create an optimal environment for storage of spermatozoa; in this regard less information is available for Acipenseridae compared with Cyprinidae and Salmonidae. The aim of the present study is to determine chemical composition and osmolality of seminal fluid and their relationship with sperm motility in Acipenser persicus. The results obtained show that sodium (Na⁺), chloride (Cl^?) and potassium (K⁺) were predominant ions in the seminal plasma and the average of osmolality of seminal plasma was 82.56 mOsm kg^?1. The higher chemical contents and osmolality compared with other sturgeon species reveal species‐specific characteristics and high secretory activity of spermatic duct in A. persicus. Significant positive correlations were observed between osmolality‐Cl^?, Na⁺‐osmolality and Na⁺–Cl^? (P<0.05, P<0.001 and P<0.05 respectively). But statistically significant correlation was not observed between seminal plasma parameters and sperm motility. Probably, the Na⁺ and Cl^? are the main electrolytes playing a major role in maintaining the osmolality of the seminal plasma and the viability of the spermatozoa in vivo.     

哈白公兔性发育期间血浆睾酮浓度、睾丸大小的变化及其与精液品质的相关性

用90日龄哈白公兔9只,每隔15d采血、测其睾丸大小(105日龄后)各1次,210日龄用假阴道每隔5d采精1次,直至试验结束。试验期为165d。在此期间还进行性活动观察。结果表明,公兔的初情期出现在158±3日龄。初情期前,公兔血浆睾酮浓度和睾丸大小平均值越大,其初情期来得越早(P<0.05)。性发育期间公兔的睾酮浓度和睾丸大小都随年龄的增长而呈上升趋势,且两者之间呈显著相关性。在初情期前后不同日龄,哈白公兔的睾酮浓度和睾丸大小与其精液品质不同指标相关显著,从而提示了早期选种的可能性。     

日粮能量、蛋白水平对辽宁绒山羊精液品质和受胎率的影响

试验研究日粮能量、蛋白质水平对种公羊精液质量和受胎率的影响。以辽宁绒山羊种公羊为试验材料,采用单因子随机试验设计,将18只成年种公羊按照采精量、产绒量平均分为3组:高能量高蛋白组(HEHP)、高能量低蛋白组(HELP)、低能量低蛋白组(LELP),每组6只。试验结果表明:日粮蛋白质水平显著影响密度、有效精子数和冻精产量;日粮能量和蛋白质水平对鲜精活力和冻精活力、鲜精畸形率、鲜精顶体完整率、母羊受胎率无显著影响。     

绵羊精浆蛋白质组2-DE图谱的构建及初步分析

本研究比较了丙酮沉淀法制备蛋白质不同上样量对绵羊精浆蛋白质二维电泳图谱的影响。结果显示,精浆总蛋白经SDS-PAGE电泳,得到分子质量在14.4~116 ku的28条蛋白质条带。二维电泳图经PDQuest 8.0分析,上样量为0.8、1.0、1.2 mg时检测出的蛋白质点分别为207±10、281±13和374±16个,分子质量基本分布在20~80 ku、等电点为4~9的区域内,随着上样量的增加,分子质量在20~80 ku的蛋白质点明显增多,但每个分子质量区间的蛋白质点所占的比率较为恒定。研究结果表明,采丙酮沉淀制备精浆蛋白结合合适的上样量能够建立绵羊精浆全蛋白质图谱,为进一步研究绵羊精浆蛋白质组学奠定基础。