绵羊睾酮和精液品质与微量元素的季节性变化

公绵羊血浆睾酮（Ｔ）水平在８～１２月极显著地高于１～７月（Ｐ＜０．０１）;平均射精量和精子活率在９～３月均显著地高于４～８月（Ｐ＜０．０１）;精子畸形率和精子数在９～１月与２～８月有极显著差异（Ｐ＜０．０１）。血浆中锌、铜、锰水平在配种季节８～１月明显地高于非配种季节的６～７月（Ｐ＜０．０１）,并与配种季节开始后Ｔ水平的升高和精液品质改善具有明显的相关性（ｒ＝０．７３２,Ｐ＜０．０１;ｒ＝０．７８２４,Ｐ＜０．０１）。     

Metabolomic differences of seminal plasma between boars with high and low average conception rates after artificial insemination

Seminal plasma is a complex biological fluid containing many metabolites including amino acids, fructose, carbohydrates and lipids Metabolites play important roles in multiple biological processes, but details and significance of the seminal plasma metabolome related to boar fertility are unknown. The aim of the present study was to compare the comprehensive metabolome of seminal plasma from boars with different conception rate after artificial insemination and to identify the potential biomarkers. Semen samples were collected from boars which divided into two groups according to the conception rates in the offspring. Seminal plasma metabolites were isolated, purified, and then subjected to Ultra-high Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-qTOF-MS) procession. A total of 576 (Positive ion mode) and 377 (Negative ion mode) metabolites were identified in seminal plasma. Metabolites were identified and categorized according to their major chemical classes, including carboxylic acids and derivatives, organooxygen compounds, amino acids, peptides, and alogues, fatty amides, fatty acyls, benzene and substituted derivatives, purine nucleotides, pyrimidine nucleotides, glycosyl compounds, fatty acids and conjugates. The results showed that 4-Aminobenzoate, Pro-Asn, Ile-Tyr, Homoveratric acid and D-Biotin were higher in semen of boar with higher conception rate (HG) versus lower conception rate (LG) (p < .05), whereas L-Serine, Butoxyacetic acid, S-Methyl-5'-thioadenosine, Capsaicin and 1-O-(cis-9-Octadecenyl)-2-O-acetyl-sn-glycero-3-phosphocholine (PAF) were lower in HG than in LG (p < .05). These metabolites may be considered as candidate biomarkers for different fertility in boars.     

金乌贼纳精囊组织结构及其精子的储存和利用

为揭示金乌贼精子进入纳精囊及产卵过程中的精子利用方式,丰富金乌贼繁殖生物学研究内容,本研究利用实验生态学和组织切片技术,检测了交配后不同时间段雌性口膜表面精子囊和纳精囊中精子数量的变化,观察分析了雌性金乌贼纳精囊的组织结构。结果显示,金乌贼纳精囊位于繁殖期雌性个体口膜腹面的突起处,共1对。纳精囊开口于口膜内表面,通过一根中央管连通整个纳精囊。中央管内壁含有大量褶皱和纤毛。在中央管两端,有12~20个储精小囊与之相连。储精小囊四周具有发达的环肌,其中储存有大量精子,并且大部分精子头部均朝向腔室内壁。完成一次交配后,雌性金乌贼对精子囊和纳精囊中精子的利用可以分为三个阶段,主要利用精子囊中的精子(交配后1~2 d);由利用精子囊中的精子向纳精囊中的精子过渡(交配后2~3 d);主要利用纳精囊中的精子(交配后3 d以上)。研究表明,从精子囊释放出的精子进入雌性口膜表面的褶皱中,通过自身运动到达纳精囊。进入纳精囊的精子通过自身运动及中央管内壁纤毛的摆动进入储精小囊,其中大部分精子头部朝向储精小囊内壁有规律地分布。在产卵过程中,雌性优先利用精子囊中的精子,而在精子囊中精子不足时,纳精囊通过肌肉收缩以及纤毛摆动将其中的精子逐渐释放出来,卵子在雌性口膜附近完成体外受精。     

非洲菊组培快繁生产中外植体诱导优化试验初报

以非洲菊试管苗叶柄切段和幼花托为外植体,进行芽诱导分化培养比较试验。结果表明,M S 6-BA 4.0m g/L IBA 1.0m g/L培养基较适合试管苗叶柄切段的诱导出芽,出芽率为30.0%~36.9%;接种后15~19d,试管苗叶柄切段的出芽率为30.0%~42.2%,污染率为0~14.7%;幼花托的诱导出芽率为0~23.1%,污染率为23.1%~69.2%,与叶柄切段差异明显。由此可见,不同外植体对诱导分化出芽的效果差异很大,以试管苗叶柄切段为外植体进行诱导分化,可缩短诱导周期,提高诱导出芽率,降低污染程度,可作为非洲菊组培快繁生产中诱导出芽的新途径。     

Application of in vitro organ culture in wide-cross breeding of rapeseed

Oil radish (Raphanus sativus var. raphanistroides Makino) is resistant to drought and low temperature. In order to breed more resistant cultivars of rapeseed, the wide cross between rapeseed (Brassica napus L.) and oil radish was made. Rapeseed was not compatible with oil radish, and the frequency of hybrid plants (F₁) was very low. Moreover, the hybrid plants were sterile. In order to recover the intergeneric hybrids (F₁), the in vitro organ culture technique was applied in our experiments. The frequency of hybrid plants (F₁) was increased up to 25.55% by means of in vitro culture of pollinated ovaries. Some fertile amphidiploid hybrid plants were obtained by means of colchicine treatment of small buds obtained from cultured flower receptacle segments of hybrid plants (F₁). It is suggested that the technique of in vitro culture of pollinated ovaries and flower receptacle segments is useful in the wide-cross breeding of rapeseed. This revised version was published online in July 2006 with corrections to the Cover Date.     

New protocol to separate llama sperm without enzymatic treatment using Androcoll-E™

The objective of this study was to design a protocol to separate spermatozoa from seminal plasma of raw llama semen without prior enzymatic treatment using a single-layer centrifugation with Androcoll-E^™ (AE). Two experiments were performed: (a) samples were divided into three aliquots (1 ml) that were deposited on the top of 4, 5 or 6 ml of AE and were centrifuged at 800g for 20 min and (b) samples were divided into two aliquots (1 ml) that were deposited on the top of 4 ml of AE and were centrifuged at 600g or 1,000g for 20 min. Columns of 5 and 6 ml of AE showed a total sperm motility (TM) significantly lower, while in the 4 ml column, this parameter was not different from the TM of samples before the AE treatment. The percentage of spermatozoa with intact and functional membranes, normal morphology and intact acrosomes, as well as the percentages of sperm with highly condensed chromatin, was conserved (p ˃ .05) in the three column heights and in the two centrifugation speeds evaluated. In conclusion, the different column heights of AE (4, 5 and 6 ml) and the different centrifugation speeds used (600, 800 and 1,000g) allow separating spermatozoa of raw llama semen without enzymatic treatment, preserving the evaluated sperm characteristics. However, of all the studied treatments, centrifugation in the 4 ml column of AE at 800g would be the method of choice to process raw llama semen samples destined for reproductive biotechnologies.     

Proteins of seminal fluid and spermatozoa in the trout (Oncorhynchus mykiss): Partial characterization and variations

The protein composition of seminal fluid, blood serum, sperm plasma membrane and flagellum of rainbow trout were analysed by SDS-polyacrylamide gel electrophoresis. Immunological identity between proteins of the 2 fluids and sperm components was studied using crossed immunoelectrophoresis, rocket immunoelectrophoresis and immunoblotting. Results indicate that many seminal proteins are antigenically-related to serum proteins, proteins of sperm origin are present in seminal fluid in varying amounts, depending on the animals and sampling time, and several serum-like seminal proteins are bound to spermatozoa. Lipoproteins were isolated from seminal fluid (mean level: 33 μg/ml) and characterized. They were identified as being HDL-like lipoproteins. A possible physiological role is proposed for these seminal lipoproteins.     

Effects of season and dietary ascorbic acid on some biochemical characteristics of rainbow trout (Oncorhynchus mykiss) semen

During the reproductive season, rainbow trout spermatozoa are stored in the sperm ducts for several months. There is no sperm production at this time since spermatogenesis is completed before spawning. To leam more about characteristics of semen during such a long storage, we analyzed changes in protein concentrations, anti-proteinase activity in seminal plasma and sperm aspartate aminotransferase activity during an extended reproductive period during which fish were fed diets supplemented with various ascorbic acid concentrations. Seminal plasma protein concentration and anti-proteinase activity declined toward the end of the reproductive season. These phenomena may be related to oncoming proteolytic events leading to degradation of the sperm. Protein concentrations and anti-proteinase activities were strongly correlated within groups of different ascorbic acid supplementations and several sampling dates (r=0.6–0.9 in most cases, p<0.05). Ascorbic acid deficiency resulted in a decrease in both parameter levels as compared to levels in groups with vitamin C supplement (p<0.08). Deficiency also resulted in lower stimulation of aspartate aminotransferase by an exogenous pyridoxal 5-phosphate in comparison to fish fed vitamin C-supplemented diets (p<0.05). These results support earlier studies suggesting a protective role of ascorbic acid toward maintaining sperm quality.     

Sperm density, seminal plasma composition and their physiological relationship in the endangered Caspian brown trout (Salmo trutta caspius)

Sperm density, mineral and organic composition of the seminal plasma and their physiological relationship were investigated in the Caspian brown trout (Salmo trutta caspius). To establish a rapid and accurate method for assessment of sperm density, three different techniques were used: sperm counting, spectrophotometry (at 480 nm) and determination of the spermatocrit. The seminal plasma contained 159.26±8.84 mM sodium (Na), 33.72±2.01 mM potassium (K), 133.04±5.96 mM chlorine (Cl), 1.68±0.2 mM calcium (Ca) and 0.988±0.13 mM magnesium (Mg). The following organic components were found: total protein 0.75±0.14 mg 100 mL⁻¹, cholesterol 2.86±0.58 mg L⁻¹ and glucose 3.81±1.04 mM L⁻¹. The mean sperm density was estimated to be 3.3 × 10⁹ spermatozoa mL⁻¹. The spermatocrit (%) ranged from 25 to 52 in sperm samples. Highly significant linear relationships were found between sperm density and spermatocrit (R²=0.703, P<0.001) and sperm density and optical density (R²=0.909, P<0.001), indicating that optical density can be used as a quick and accurate method of estimating sperm density. Significant relationships were also found between sperm density and Ca, Mg and total protein of seminal plasma. A significant correlation was also observed between the Ca and Mg concentrations (R²=0.774, P<0.01). The following correlations were observed between mineral and organic components: total protein and Ca (R²=0.462, P<0.05), total protein and Mg (R²=0.518, P<0.05) and glucose and Cl (R²=0.374, P<0.05). These parameters should be considered when developing procedures for either artificial fertilization or for cryopreservation of sperm.     

Separation and Purification of Ovulation-inducing Factors in the Seminal Plasma of the Bactrian Camel (Camelus bactrianus)

Ovulation in the Bactrian camel depends upon ovulation-inducing factors in the seminal plasma. The present study was conducted to isolate and purify the bioactive fractions from the seminal plasma of these camels. The seminal plasma was fractionated by anion-exchange chromatography, and six fractions were obtained. The bioactive potential of each fraction was estimated from its effect on rat pituitary tissue cultured in vitro and by the effect of an intramuscular injection of the fraction into female camels in vivo. Both the third fraction (F3) and the fifth fraction (F5) stimulated the release of LH in vitro and in vivo. In addition, female camels ovulated within 48 h after intramuscular injection of F3. However, neither F3 nor F5 had any significant effect on the secretion of FSH, either in vitro or in vivo. When F3 was further fractionated into four subfractions, the third subfraction (F3-3) still stimulated the in vitro release of LH, but not of FSH. An attempt to further purify the ovulation-inducing factors in F3-3 failed owing to the similarity of the molecular characters.