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.     

The Concentrations of LH,FSH, Oestradiol-17β and Progesterone in the Blood Plasma of the Female Bactrian Camel (Camelus bactrianus) before and after Intramuscular Injection of Seminal Plasma

The concentrations of LH, FSH, oestradiol-17beta and progesterone in the blood plasma of the female Bactrian camel (Camelus bactrianus) before and after intramuscular injection of seminal plasma.     

双峰驼精清及其活性组分凝集素寡糖的初步研究

应用植物血凝素与寡糖结构相同或相似的糖复合物专一凝集的原理与方法,结合小鼠排卵试验,间接观察了公驼精清及其经ＤＥ－５２离子交换层析和高效液相层析提取分离的活性组分的蛋白组成。结果表明,公驼精清内糖蛋白含有２种以上的寡糖,其内的活性组分及诱导排卵因子与所试植物血凝素不发生反应     

哺乳动物精清诱导排卵活性蛋白质研究

在动物的精清里包含一些与动物生殖生理活动密切相关的活性蛋白和多肽。驼科动物的双峰驼、美洲驼、羊驼和澳洲考拉等哺乳动物精清中含有类似促性腺激素释放激素(GnRH)作用的活性蛋白质,诱导排卵因子(ovulation inducing factor,OIF),它能引发外周血液中促黄体素(LH)峰浓度的出现和诱发母畜排卵。OIF被认为是一种新的生物活性蛋白质或潜在的新型激素。甚至报道在牛等自发排卵动物以及人精清中也存在类似的活性蛋白。这类因子的发现对于哺乳动物生殖生理及繁殖的深入研究具有重要的意义,有助于动物克服繁殖障碍及繁殖疾病的治疗等。     

Effects of a single injection of GnRH or equine pituitary extract on plasma LH and FSH concentrations in stallions

Plasma LH and FSH concentrations were measured in mature stallions after administration of synthetic GnRH or equine pituitary extract. GnRH caused significant rises in plasma LH (2-fold) and FSH (1.7-fold). Concentrations of LH remained significantly elevated for 4 hours and FSH remained elevated for 2 hours. Similar increases in plasma LH (1.6-fold) and FSH (1.8-fold) occurred after an injection of equine pituitary extract. LH was significantly elevated for 4 hours and FSH was elevated for 6 hours.     

Plasma Inhibin Levels in Relation to Steroids and Gonadotrophins during Oestrous Cycle in Buffalo

The study was conducted on six Murrah buffalo synchronized and induced to oestrus. An indwelling catheter was placed in the jugular vein of each buffalo 4 days before the expected onset of the oestrus following the induced oestrus and blood samples were collected at 8 h intervals from each animal throughout the oestrous cycle. Plasma immunoreactive inhibin, follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol-17β and progesterone were estimated by radioimmunoassay to study the variations in the peripheral levels of these hormones and their inter-relationships in order to elucidate the feedback systems controlling them during the oestrous cycle of buffalo. Plasma inhibin levels ranged between 391.25 and 631.97 pg/ml during various phases of the oestrous cycle and were found to be higher than reported in cows. Peak LH and FSH levels during oestrus were 38.40 ± 9.21 and 24.04 ± 4.75 ng/ml, respectively and estradiol-17β and progesterone were 19.50 ± 5.51 pg/ml and 0.61 ± 0.25 ng/ml, respectively. The mean plasma inhibin concentration on the day of oestrus was 562.5 ± 18.9 pg/ml. Levels of FSH in the plasma showed three mid-cycle elevations which corresponded to comparatively lower inhibin and elevated estradiol-17β levels during the same period. From this observation it was deduced that both inhibin and estradiol-17β have a feed-back regulatory effect on FSH secretion in buffalo.     

Preliminary Observations on the Concentrations of Serum Luteinizing Hormone and Follicle-stimulating Hormone during Gestation in Bactrian Camels (Camelus bactrianus)

The concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the serum of the Bactrian camel (Camelus bactrianus) throughout pregnancy were analysed by radioimmunoassay. The samples were withdrawn at intervals of half a month. The serum LH concentrations varied significantly between 2.2±0.9 and 20.3±18.8 ng/ml during the pregnancy, with the highest value on the day of artificial insemination, which indicated that a preovulatory LH peak appeared. Thereafter, the LH concentration tended to fall, reaching a fairly constant low level by the eighth month of pregnancy. The serum FSH concentrations varied between 7.0±0.2 and 28.9±0.4 ng/ml during gestation, with the peak value being reached at 4.5 months, followed by a marked drop to 7.0±0.2 ng/ml at 7.5 months.     

猪垂体细胞单层培养

本研究建立了猪垂体前叶细胞单层培养模型 ,并进行了免疫细胞化学研究。结果显示 ,用 0 2 5 %胶原酶 ,0 0 5 %透明质酸酶和 0 0 1%DNA酶消化垂体前叶 2次 ,每个垂体可获 5× 10 8个细胞 ,细胞活率在 90 %以上。用含 10 %小牛血清的McCoy’s 5A培养液培养 2 0h ,细胞贴壁 ,4 0h左右形成单层 ,形成单层后换无血清培养液仍维持生长。抗LH和FSH免疫细胞化学染色表明 ,大多数LH阳性细胞同时也是FSH阳性细胞。在单层细胞中 ,LH和FSH阳性细胞占细胞总数的比例不足 10 %。本培养模型可用于体外研究垂体细胞的功能及调控     

日粮能量对乌鸡外周血清促性腺激素与垂体分泌促性腺激素的影响

采用体外细胞培养和放射免疫测定法（RIA）测定日粮能量对鸟鸡外周血清促性腺激素与垂体分泌促性腺激素的影响。日粮能量处理：低能Ⅱ组、低能I组、对照组、高能I组、高能Ⅱ组;体外培养垂体细胞组分：高能组、对照纽、低能组。结果显示,血清中FSH含量,对照组与高能Ⅱ组相比差异显著（P〈0．05）,与低能I、Ⅱ组相比差异极显著（P〈0．01）;血清中LH含量,对照组与高能I组、高能Ⅱ组组间差异显著（P〈0．05）;低能Ⅱ组与高能I组、低能Ⅱ组与高能Ⅱ组、低能I组与高能I组、低能I组与高能Ⅱ组组间差异极显著（P〈0．01）;单层垂体细胞中FSH含量,高能组和对照组与低能组比差异显著（P〈0．05）;单层垂体细胞中LH含量,高能组与低能组比较差异极显著（P〈0．01）,对照组与低能组比较差异不显著。结果表明,日粮能量对处理体外培养乌鸡垂体细胞分泌FSH、LH有促进作用。     

Incubation of Post-Thaw Epididymal Cat Spermatozoa with Seminal Plasma

In domestic cats, epididymal spermatozoa have lower initial motility and viability than ejaculated spermatozoa and it is possible that seminal plasma compounds are behind these effects. The aim of this study was to investigate whether co-incubation of post-thaw epididymal cat spermatozoa with seminal plasma was able to improve sperm quality. Epididymal cat spermatozoa from 11 cats were cryopreserved. After thawing, each sperm sample was divided into two aliquots, centrifuged and incubated with two different media; Tris buffer (control) or pooled seminal plasma (treatment). Sperm quality was observed at 0, 2, 4 and 6 h after incubation. The results demonstrated that all of the sperm parameters except acrosome integrity were lower in the treatment group compared to the control group (p < 0.05); the percentages of motility (46.4 ± 15.4 vs 40.0 ± 9.4), the scores of progressive motility (3.1 ± 0.4 vs 2.8 ± 0.5), the percentages of spermatozoa with intact plasma membrane (46.3 ± 9.7 vs 39.6 ± 8.9) and intact acrosome (36.5 ± 16.2 vs 32.9 ± 15.1), as well as at all time points. In conclusion, the seminal plasma seems less beneficial to the post-thaw epididymal cat spermatozoa than the Tris buffer.     

初情期前母猪垂体细胞LH和FSH的释放及其对GnRH类似物的反应

利用垂体细胞单层培养模型研究了30，60，90，120和150日龄北京黑猪母猪垂体细胞LH和FSH的释放及其对LRH－A3反应能力。结果表明，猪垂体细胞LH和FSH释放对LRH－A3的反应呈S形剂量依赖型同线。LH基础释放量和LRH－A3刺激的最大释放量在不同日龄间没有显著差异，说明初情期前母猪垂体细胞已达最大LH释放和对GnRH反应的能力。FSH的基础释放量和LRH－A3刺激的最在释放量在30日龄与60日龄之间无差异，但90日龄以后垂体细胞FSH的释施量随日龄增大而降低，提出可能由于垂体在体内时受过抑制素的作用。     

Effect of LPS on Reproductive System at the Level of the Pituitary of Anestrous Ewes

In our research we focused our attention on the effect of the immune stress induced by bacterial endotoxin–lipopolysaccharide (LPS) on the hypothalamic–pituitary–gonadal axis (HPG) at the pituitary level. We examined the effect of intravenous (i.v.) LPS injection on luteinizing hormone (LH) and follicle‐stimulating hormone (FSH) release from the anterior pituitary gland (AP) in anestrous ewes. The effect of endotoxin on prolactin and cortisol circulating levels was also determined. We also researched the effect of immune challenge on the previously mentioned pituitary hormones and their receptors genes expression in the AP. Our results demonstrate that i.v. LPS injection decreased the plasma concentration of LH (23%; p < 0.05) and stimulates cortisol (245%; p < 0.05) and prolactin (60%; p < 0.05) release but has no significant effect on the FSH release assayed during 6 h after LPS treatment in comparison with the control levels. The LPS administration affected the genes expression of gonadotropins’β‐subunits, prolactin and their receptors in the AP. Endotoxin injection significantly decreased the LHβ and LH receptor (LHR) gene expression (60%, 64%; p < 0.01 respectively), increased the amount of mRNA encoding FSHβ, FSH receptor (FSHR) (124%, 0.05; 166%, p < 0.01; respectively), prolactin and prolactin receptor (PRLR) (50%, 47%, p < 0.01; respectively). The presented, results suggest that immune stress is a powerful modulator of the HPG axis at the pituitary level. The changes in LH secretion could be an effect of the processes occurring in the hypothalamus. However, the direct effect of immune mediators, prolactin, cortisol and other components of the hypothalamic pituitary–adrenal (HPA) axis on the activity of gonadotropes has to be considered as well. Those molecules could affect LH synthesis directly through a modulation at all stages of LHβ secretion as well as indirectly influencing the GnRHR expression and leading to reduced pituitary responsiveness to GnRH stimulation.     

Hormonal Response of Female Goats to Active Immunization against a Recombinant Human Inhibin α-subunit, and establishment of an Enzyme-linked Immunosorbent Assay for Caprine Follicle-stimulating Hormone

The effect of selective immunosuppression of endogenous inhibin in goats on FSH, LH, progesterone and estradiol-17β profiles was studied during the breeding and nonbreeding seasons. Eighteen adult female Boer goats were immunized against the recombinant human inhibin α-subunit (hINH-α). With the exception of estradiol, which was determined by radio-immunoassay (RIA), all plasma hormone concentrations were determined by ELISA. The ELISA for FSH presented in this paper was established in the authors' laboratory, based on an existing RIA. Mean basal concentrations of FSH were not affected by immunosuppression of endogenous inhibin, nor was there a difference in the amplitude of the pre-ovulatory FSH surge. Immunization against inhibin appears to eliminate the slight secondary rise of FSH occurring 12–20 h after the major surge associated with ovulation. The LH profiles of the immunized goats were characterized by lower basal concentrations both before and after the pre-ovulatory LH surge which itself was reduced by 50% in immunized does. By contrast, concentrations of circulating estradiol were significantly elevated after inhibin-immunization. Progesterone profiles were not affected. Extending immunization into the anoestrous season by a booster injection of hINH-α, implicating oestrus induction with a progestagen and eCG, produced no discernible differences in FSH and LH profiles in comparison with nonimmunized control goats. The findings suggest that in goats, paracrine factors may play a more significant role in controlling follicular activity than a feedback mechanism acting via the pituitary.     

Plasma pharmacokinetics and urine concentrations of meropenem in ewes

The pharmacokinetics of meropenem was studied in five ewes after single i.v. and i.m. dose of 20 mg/kg bw. Meropenem concentrations in plasma and urine were determined using microbiological assay method. A two-compartment open model was best described the decrease of meropenem concentration in plasma after an i.v. injection. The drug was rapidly eliminated with a half-life of elimination ( t _{1/2 β} ) of 0.39 ± 0.30 h. Meropenem showed a small steady-state volume of distribution [ V _d(ss)] 0.055 ± 0.09 L/kg. Following i.m. injection, meropenem was rapidly absorbed with a t _1/2ab of 0.25 ± 0.04 h. The peak plasma concentration ( C _max) was 48.79 ± 8.83  μ g/mL was attained after 0.57 ± 0.13 h ( t _max). The elimination half-life ( t _1/2el) of meropenem was 0.71 ± 0.12 h and the mean residence time ( MRT ) was 1.38 ± 0.26 h. The systemic bioavailability (F) after i.m. injection was 112.67 ± 10.13%. In vitro protein-binding percentage of meropenem in ewe's plasma was 42.80%. The mean urinary recoveries of meropenem over 24 h were 83% and 91% of the administered dose after i.v. and i.m. injections respectively. Thus, meropenem is likely to be efficacious in the eradication of many urinary tract pathogens in sheep.     

Distinct Effects of Boar Seminal Plasma Fractions Exhibiting Different Protein Profiles on the Functionality of Highly Diluted Boar Spermatozoa

The aim of this study was to evaluate how different protein profiles of seminal plasma (SP) fractions affect sperm functionality in vitro. Ejaculates from three boars were separated into six fractions. The fractions differed from each other in their sperm content, in their total SP protein content, and their spermadhesin PSP-I/PSP-II and heparin-binding protein (HBP) concentrations. Spermatozoa were mainly recovered in fraction 2 (sperm-rich fraction, >1800 × 10⁶ spermatozoa/ml), whereas the pre-sperm fraction 1 and the post-sperm fractions 4–6 contained low numbers of spermatozoa (<500 × 10⁶/ml). Except in fraction 2, the total SP protein concentration and the concentration of both, spermadhesin PSP-I/PSP-II and the HBPs increased with fraction order. Distinct time-dependent effects were observed on motility characteristics and membrane integrity of highly diluted boar spermatozoa upon incubation with a 10% dilution of the SP from each fraction. The highest sperm viability was recorded after exposure for 5 h to fraction 2, followed by fractions 1 and 3. The percentages of motile spermatozoa also differed significantly among fractions after 5 h of incubation. Spermatozoa incubated with SP of fractions 1–3 showed the highest percentage motility. We conclude that different SP fractions exert distinct effects on the functionality of highly diluted boar spermatozoa. Fractions 1–3 appear to promote sperm survival, whereas fractions 4–6 seem to be harmful for preserving the physiological functions of highly diluted boar spermatozoa.     

复方中草药添加剂对辽宁绒山羊种公羊繁殖性能的影响

对6只3.79岁±0.29岁的辽宁绒山羊种公羊试验组羊饲喂复方中草药添加剂,研究较高采精强度下中草药添加剂对种公羊繁殖性能的影响。结果表明,饲喂60 d后试验组种公羊射精量、精子密度极显著高于对照组(P<0.01),顶体完整率显著高于对照组(P<0.05);美兰褪色时间极显著低于对照组(P<0.01)。试验组血浆和精浆FSH、LH和T略高于对照组,但差异不显著(P>0.05);相关分析发现,试验组血浆T(y,ng/L)与血浆FSH(x,u/L)呈对数关系;试验组血浆T(y,ng/L)与血浆LH(x,μg/L)也呈对数关系,并与对照组的相关方程近似。试验组精浆T含量(y,ng/L)与LH(x,μg/L)之间也存在对数相关关系,并与对照组的相关方程相似。表明在该研究条件下,在较高采精强度下的种公羊按照15 g/d的剂量饲喂复方中草药添加剂,使参试动物的精液数量和质量均得到改善,对血浆和精浆生殖激素影响很小,维持了绒山羊的生殖内分泌平衡状态。因此,以淫羊藿、巴戟天和枸杞子为主的复方中草药添加剂对改善种公羊的繁殖性能效果显著。     

山羊GnRH和促性腺激素的释放特点

通过外科手术分别连续收集活体山羊中黄体期及早卵泡期的垂体门脉血样和外周血样，经放射免疫测定，山羊中黄体期和早卵泡期的促性腺激素释放激素（ＧｎＲＨ）、促黄体生成素（ＬＨ）和促卵泡素（ＦＳＨ）均呈波动式释放。在早卵泡期，ＦＳＨ单位时间内波动次数和血浆平均水平显著高于中黄体期；ＧｎＲＨ与ＬＨ的波动型基本一致，ＦＳＨ的变化不太规则。表明山羊垂体促性腺激素的释放受丘脑下部ＧｎＲＨ的调节，但ＦＳＨ似乎还存在其他调节机理。     

阿拉善双峰驼及驼乳、驼肉研究概述

阿拉善双峰驼是世界主要骆驼品种之一,是宝贵的骆驼种质资源。文章综述了阿拉善双峰驼的驯养历史和驼乳、肉产品的研究概况,结合调研和测定数据分析了驼肉中氨基酸的成分及含量,以期为保护阿拉善双峰驼种质资源以及深入开发双峰驼系列产品提供帮助。     

日粮能量对育成柴鸡外周血浆促性腺激素与垂体分泌促性腺激素的影响

采用单因素方差随机试验,选用14周龄育成柴鸡36只,随机的分成3个处理,每个处理3个重复,每个重复4只鸡,分别饲喂低能、对照、高能的日粮,试验期为28d。结果显示,育成柴鸡外周血浆FSH浓度,低能、对照与高能差异极限著（P〈0.01）,并且随着日龄增加FSH浓度逐渐增加。育成柴鸡外周血浆LH浓度第1周时低能组与对照、高能组差异极显著（P〈0.01）,对照与高能组差异不显著;第2、3周处理低能组、对照、高能组差异极显著（P〈0.01）,并且随着日龄增加浓度增加。育成柴鸡体外培养垂体细胞分泌FSH、LH高能组与低能、对照组差异极显著（P〈0.01）,而低能组与对照组差异不显著。结果表明,日粮能量通过下丘脑-垂体-性腺轴影响育成柴鸡生殖机能。     

Concentrations of bioactive and immunoactive luteinizing hormone in bovine anterior pituitary tissue

Tissue concentrations of LH as determined by radioimmunoassay (RIA) may not accurately measure actual changes that could occur in biological activity of the hormone. To examine this possibility, pituitary homogenates from 135 beef cows in various physiological states were analyzed for content of LH by both RIA and an in vitro bioassay. The ratio of biological/immunological active concentrations of LH remained constant (.52 +/- .02) even though tissue concentrations of immunoactive LH differed among groups. Tissue concentrations of bioactive LH were linearly related to, and highly correlated with (P less than .001), tissue concentrations of immunoactive LH. These data indicate that only a fraction of the immunoactive LH in the bovine pituitary is biologically active. However, this fraction does not vary with the reproductive status or plane of nutrition.