广聚萤叶甲雄虫生殖系统特异性精液蛋白基因的筛选及表达分析

为明确广聚萤叶甲Ophraella communa雄虫精液蛋白基因在性别间的表达差异并筛选出在雄虫生殖系统特异性表达的基因，从转录组及蛋白组数据库选取13个精液蛋白基因，采用实时荧光定量PCR技术检测其在广聚萤叶甲不同性别、组织及发育阶段的表达特征。结果显示，在广聚萤叶甲雄虫13个精液蛋白基因中，有9个精液蛋白的mRNA表达水平在雄虫中更高，有5个精液蛋白基因具有专一的雄虫生殖系统特异性，分别为PGK、sucA、ENO、SDH和PLA2b基因，在雄虫生殖系统中的表达量分别较其在头部的表达量高273.01倍、401.48倍、11.21倍、64.27倍和1.86倍。PGK、sucA和ENO基因均在广聚萤叶甲老熟蛹期急剧上调表达，在性成熟成虫中的表达量最高，分别是卵期表达量的43.48倍、31.03倍和11.89倍，三者具有明显的组织和发育阶段时空特异性，推测PGK、sucA和ENO基因参与广聚萤叶甲雄虫生殖的可能性较其余基因更高。     

A comparative study of the effects of intramuscular administration of gonadorelin,mating and intrauterine infusion of either raw seminal plasma or seminal plasma purified β‐NGF on luteal development in llamas

The aim of this study was to compare the effect of the intramuscular administration of 50 μg of gonadorelin acetate versus natural mating, intrauterine infusion (i.u.) of a physiological relevant dose of either raw llama seminal plasma (SP) or purified beta‐nerve growth factor from seminal origin (spβ‐NGF) on ovulation rate and corpus luteum (CL) development and function in llamas. Females with a follicle (≥8 mm) were assigned to groups: (i) i.m. administration of 50 μg of gonadorelin acetate (GnRH; positive control; n = 4); (ii) single mating (mating; n = 6); (iii) i.u. infusion of 4 ml of llama SP (SP; n = 4); or (iv) i.u. infusion of 10 mg of spβ‐NGF contained in 4 ml of PBS (phosphate‐buffered saline) (spβ‐NGF; n = 6). Ovaries were examined by power Doppler ultrasonography at 0, 1, 3, 6, 12 and 24 hr after treatment to determine preovulatory follicle vascularization area (VA), and additionally every 12 hr until Day 2 (Day of treatment = Day 0) to determine ovulation. Afterwards, ovaries were examined every other day until Day 8 to evaluate CL diameter and VA. Blood samples were collected on Days 0, 2, 4, 6 and 8 to determine plasma progesterone (P4) concentration. Ovulation rate did not differ (p = .7) among groups, but treatment affected (p < .0001) preovulatory follicle VA. Neither treatment administration nor treatment by time interaction affected (p ≥ .4) CL diameter, VA and plasma P4 concentration. Mating tended (p = .08) to increase CL VA when compared to the seminal plasma group by Day 8. Intrauterine administration of seminal plasma or spβ‐NGF does not increase CL size and function when compared to i.m. GnRH treatment, suggesting that the administration route of spβ‐NGF influences its luteotrophic effect in llamas.     

Serum testosterone,sperm quality,cytological, physicochemical and biochemical characteristics of the prostatic fraction of dogs with prostatomegaly

Prostatomegaly is a common finding in older non‐neutered dogs. This study compared the serum testosterone, sperm quality and characteristics of the prostatic fraction between healthy dogs and dogs with prostatomegaly. Blood samples of ten dogs (five dogs from each group) were taken for serum testosterone measurement. Sperm motility, vigour, concentration, viability, membrane functionality and morphology were analysed in sperm‐rich fraction. Osmolality, pH, cell types, and albumin, haemoglobin, acid phosphatase, alkaline phosphatase, glucose, triglycerides, cholesterol, calcium, phosphorus, magnesium and chloride were analysed in prostatic fraction. Dogs with prostatomegaly have the lowest sperm motility, vigour, concentration and functional membrane. Dogs with prostatomegaly have the highest glucose, triglycerides and cholesterol. Glucose was the only constituent positively correlated with serum testosterone and prostate volume. It can be concluded that dogs with prostatomegaly have poorer sperm quality, and glucose, triglycerides and cholesterol in prostatic fraction can be used as prostatomegaly biomarkers.     

Sperm characteristics of chub Leuciscus cephalus (L.) collected in artificial condition after Ovopel and Ovaprim treatment

The effect of two commercial preparations containing different GnRH analogues with dopamine antagonists on quantitative and qualitative parameters of semen from chub Leuciscus cephalus L. collected in artificial conditions were examined. Semen was collected after the application of [(D‐Ala⁶, Pro⁹ NEt)‐mGnRH + metoclopramide] (Ovopel, n = 9), [(D‐Arg⁶, Pro⁹ Net)‐sGnR + domperidone] (Ovaprim, n = 9) and from the control group (0.9% NaCl, n = 9). Afterwards, semen volume, sperm concentration, total sperm production and semen pH were determined. Osmolality and pH of seminal plasma were also determined. Using the Computer‐assisted sperm analysis system (CASA), selected sperm parameters such as sperm motility (MOT %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, μm s^?1), straight‐line velocity (VSL, μm s^?1), movement linearity (LIN, %), wobbling index (WOB, %), amplitude of lateral head displacement (ALH, μm) and beat cross frequency (BCF, Hz) were analysed. While Ovopel can also be used to stimulate chub spermation, the application of Ovaprim was much more effective for obtaining higher amounts of semen.     

牛精清及牛用稀释液在猪精液冷冻中的应用

为了验证牛精清及牛用精液冷冻稀释液在猪精液冷冻中的有效性,将牛精清(0、2.50%、5.00%、10.00%)加入猪精液冷冻稀释液(葡萄糖1.10%、柠檬酸1.48%、Tris 2.42%、NAC 0.02%、卵黄20.00%、甘油6.00%、青霉素0.06%、链霉素0.10%)中,或用改良猪精液冷冻稀释液(另加入0.10 mmol/L他克林)、牛精液冷冻稀释液及改良牛精液冷冻稀释液,对猪精液进行冷冻处理。结果表明,在本试验条件下,牛精清对猪精子冷冻保存没有正面作用;改良牛用稀释液用于猪的精液冷冻,效果与改良猪用稀释液相当,优于未改良牛用稀释液。     

精浆对猪精子冷冻效果影响的研究

为优化猪精子冷冻技术,提高解冻后精子的活力和受精能力,本试验分别以含精浆浓度为10%、20%和30%的冷冻保护剂处理精子,以冷冻前、解冻后精子活力和质膜完整性,解冻后精子进行体外受精(IVF)的卵裂率和囊胚率等作为检测指标,同时以含有卵黄的Tris-柠檬酸－葡萄糖(Tris-citric acid-glucose,TCG)冷冻基础液作为对照研究精浆对猪冷冻精子的保护作用。结果显示,在含有10%精浆浓度的稀释液中,冷冻前质膜完整性,解冻后精子活率、质膜完整性、IVF囊胚率相对于对照组均显著提高(P<0.05);当含有10%精浆的冷冻精液解冻后用于人工授精时,与配母猪妊娠率、窝产仔数、窝产活仔数等仍显著低于鲜精授精组(P<0.05)。上述结果表明,含10%精浆的冷冻保护剂能提高精子的冷冻后活力和IVF胚胎发育率,但用于人工授精配种与鲜精相比还有一定差距。     

气-质联用和酶联免疫检测草莓花托中ABA含量

以草莓＂杜克拉＂为试材,分别利用ELISA和GC-MS测定草莓花托中4个时期ABA含量。结果表明,ELISA（ng/g）/GC-MS（ng/g）检测草莓大绿期（大绿）、白色期（纯白）、着色期（始红）和始熟期（片红）花托中ABA含量分别为65.93/72.90,74.49/233.55,69.56/172.11和78.26/225.07。除大绿果期外,其他时期GC-MS检出结果极显著高于ELISA,但二者变化趋势基本相同,即呈N型升-降-升趋势,这表明ABA与草莓果实成熟存在密切关系。通过对造成差异原因、操作程序复杂性和成本比较分析后认为,GC-MS虽然测量精度高,但如果研究ABA的变化趋势,可选择成本低、操作简便、安全的ELISA方法。     

双峰驼精清诱导排卵因子的研究进展

双峰驼具有独特的繁殖生理机能，是季节发情、诱导排卵的动物。文章对双峰驼繁殖特性和卵泡发育及诱导排卵因子的发现、分离和纯化、部分生物学性质等方面的研究进展进行了简要综述。     

Concentrations of minerals in the canine prostatic fluid

Current knowledge about the composition of the prostate fluid in healthy male dogs is limited and restricted to small case numbers. Furthermore, published data often vary significantly regarding sample processing and analytical methods. Therefore, we aimed to provide data on the composition of electrolytes and minerals in the canine prostatic fluid in a larger population (n = 30 dogs/samples) and to compare these results with the existing literature. Concentrations of sodium, potassium and copper analysed in our population were most consistent with those in the literature. Different to this, concentrations of total calcium, magnesium, zinc and inorganic phosphate varied. Whereas magnesium, zinc and inorganic phosphate seemed to depend on the analysis method, total calcium concentrations differed if centrifugation was performed or not. Our results clearly indicate a need for standardization of methods for analysis of seminal plasma components.     

Extenders modify the seminal plasma ability to minimize freeze‐thaw damage on ram sperm

Seminal plasma (SP) proteins interact with sperm plasma membrane (PM) modulating its functionality. It has been shown that SP proteins can reverse the damage caused by freeze‐thaw; however in these studies, SP has been added to washed sperm (i.e., cells depleted from homologous SP and extender). The aim of the current study was to assess whether the egg yolk‐based extender (EY) modifies SP ability to ameliorate sperm parameters in frozen‐thawed ram spermatozoa. Ejaculates were diluted in EY or soybean lecithin‐based extender (SL) and evaluated before and after freezing to measure the cell damage according to the extender. Even when all classical parameters decreased after freezing, as expected (p < .05), there was no effect of the extender. SP treatment was applied after freeze‐thaw. Sperm were incubated with SP (20% v/v) in the presence of either EY or SL, and sperm parameters were assessed after thawing compared with the same treatments after Percoll sperm selection (washed). Treatments with 20% SP improved sperm total and progressive motility compared with controls regardless of washing and extender (p < .05); however, washed sperm showed higher percentage of total sperm motility compared with those unwashed (p < .05). Moreover, treatment with 20% SP showed significantly higher percentages of PM integrity, sperm with intact acrosomes, integrity of chromatin and non‐capacitated sperm in samples diluted with EY when washed before treatment compared with the other conditions (p < .05). It was concluded that the presence of the extenders and particularly egg yolk alters the SP capacity to reduce the cryodamage.