Reproductive Development of Santa Inês Rams During the First Year of Life: Body and Testis Growth,Testosterone Concentrations,Sperm Parameters,Age at Puberty and Seminal Plasma Proteins

We have investigated the reproductive development of the tropically adapted Santa Inês ram, the most common hair sheep in Brazil. From 8 to 48 weeks of age, 16 animals were evaluated for body and testis growth, semen parameters, testosterone concentrations and seminal plasma proteins, using two‐dimensional SDS‐PAGE. Animals were weaned at 30 days and kept in feedlots thereafter, receiving hay, concentrate (18% of crude protein) and mineral supplement. Body weight increased from 12.3 ± 0.7 to 54.3 ± 1.6 kg between 8 and 48 weeks (p < 0.05), but changes in thoracic perimeter and scrotal circumference were non‐significant after 36 weeks (p > 0.05). The percentage of motile sperm increased slowly until 23 weeks and more rapidly after that age, but significant changes in progressive motility occurred after 25 weeks. Presence of abnormal sperm related inversely to age. Most significant changes in sperm concentration occurred between 38 and 44 weeks (0.38 ± 0.05 to 1.14 ± 0.24 × 10⁹ cells/ml, p < 0.05) and testosterone reached its highest concentrations at 42 weeks, decreasing afterwards. Rams reached puberty at 28.2 ± 0.8 weeks. The number of protein spots on seminal plasma gels was similar from 15 to 18 weeks (45 and 47 spots; p > 0.05), increased until 24 weeks (141 spots) and 28 weeks (170 spots; p < 0.05) and remained without significant (p > 0.05) changes from 28 to 48 weeks (186 ± 10 spots). Furthermore, the intensity of selected spots on 2D maps increased (p < 0.05) between 15 and 28 weeks, which preceded or coincided with the main developmental changes in sperm motility and percentage of defective sperm in the ejaculates. These results will support future studies designed to characterize specific seminal plasma proteins whose expression relate to the development of testis, epididymis and accessory sex glands.     

Effects of Melatonin Implants During Non‐Breeding Season on Sperm Motility and Reproductive Parameters in Rasa Aragonesa Rams

The effect of melatonin implants administered during non‐breeding season in Rasa Aragonesa rams on sperm motility parameters and other reproductive traits was assessed. In a first experiment, two Rasa Aragonesa rams were implanted (with melatonin group M), remaining other two males as control group (C). Semen of each group was collected from 1 May to 23 June, twice or three times a week, and motility parameters were assessed using a computer‐assisted sperm analysis system. Melatonin increased the percentage of progressive motile spermatozoa, particularly during 46–75 days after melatonin implantation (p < 0.01). In experiment 2, M and C in vitro fertilization ability had been determined by zona‐pellucida binding assays, using spermatozoa from experiment 1, obtained 60–70 days after melatonin was implanted. A significantly higher number of spermatozoa attached per oocyte was observed in frozen‐thawed immature ovine oocytes incubated with sperm from M animals than in those incubated with sperm from the C group (p < 0.01). Finally, a field assay (experiment 3) was performed. In this case, five Rasa Aragonesa rams were implanted with melatonin and three remained as control group. Sperm doses from those animals were used for artificial insemination of 2608 Rasa Aragonesa ewes from 39 different farms at non‐breeding season. Fertility, litter size and fecundity were studied. Semen from melatonin implanted rams seemed to increase both fertility and fecundity in ewes inseminated with spermatozoa obtained 46–60 days after implantation (p < 0.1). Thus, melatonin treatment in rams during non‐breeding season modifies sperm motility parameters and seems to improve the fertilization parameters obtained.     

Study of Sperm Concentration,Seminal Plasma Composition and their Physiological Correlation in the Persian Sturgeon,Acipenser persicus

The objectives of the present study were to determine ionic and organic composition of seminal plasma, sperm concentration and their relationships in the Persian sturgeon (Acipenser persicus). In this regard, ionic content (Na⁺, K⁺, Cl^?, Ca²⁺ and Mg²⁺) and organic content (total protein, glucose, cholesterol and triglyceride) along with sperm concentration were measured in 17 specimens of the Persian sturgeon. The seminal plasma contained 59.53 ± 2.56 mm /l sodium, 9.1 ± 1.42 mm chloride, 4.72 ± 0.3 mm potassium, 1.45 ± 0.075 mm calcium and 0.7 ± 0.072 mm magnesium. The following organic contents were found: total protein 0.11 ± 0.02 g/dl, glucose 22.18 ± 4.16 mg/dl, cholesterol 6.67 ± 1.04 mg/dl and triglyceride 15.2 ± 0.65 mg/dl. The mean sperm concentration was estimated to be 1.6 ± 0.12 (×10⁹ sperm/ml). A significant relationship was found between sperm concentration and K⁺ of seminal plasma (r = 0.533, p < 0.05). Significant correlations were observed between ionic contents: Na⁺ vs Cl^? (r = ?0.854, p < 0.01) and Mg²⁺ vs K⁺ (?0.583, p < 0.05). Also, level of triglyceride was negatively correlated with Mg²⁺ (r = ?0.503, p < 0.05). Presented data could be considered as a complementary study for developing special extenders and protectant solutions for improving artificial fertilization in this valuable species.     

Effect of Sperm Cryopreservation and Supplementing Semen Doses with Seminal Plasma on the Establishment of a Sperm Reservoir in Gilts

Frozen-thawed (FT) boar sperm have a reduced fertile life, due in part to a capacitation-like status induced by cooling. Reversal of this cryocapacitation in vitro by exposure to boar seminal plasma (SP) has been demonstrated. The objective of these studies was to determine the effect of SP on the ability of FT sperm to create an oviductal sperm reservoir following artificial insemination (AI). In Experiment one, 35 pre-pubertal gilts were injected (IM) with 400 IU eCG plus 200 IU hCG to induce oestrus. At detection of oestrus, gilts were inseminated with 3 x 10(9) live sperm, either fresh (FS; n = 13), FT (n = 10), or FT supplemented with 10% v/v SP (n = 12). Gilts were killed 8 h later, their reproductive tracts recovered and the uterotubal junctions (UTJs) flushed to recover sperm. Fewer (p < 0.01) sperm were recovered following FT, compared to FS, inseminations, and there was no evident effect of SP. In Experiment two, 30 pre-pubertal gilts received IM injections of 1000 IU eCG followed by 5 mg pLH 80 h later to control time of ovulation. Gilts were inseminated with 3 x 10(9) live FS sperm (n = 6), FT sperm (n = 15) or FT sperm plus 10% SP (n = 9) at 12 h before ovulation and then sacrificed 8 h later. The UTJs were dissected and flushed for sperm recovery. Fewest (p < 0.001) sperm were recovered following FT insemination and there was no evident effect of SP. These data demonstrate that the size of the sperm reservoir is markedly reduced in gilts inseminated with FT sperm. However, the lack of effect of SP suggested that either it did not reverse cryocapacitation or that such a reversal does not impact the in vivo ability to create a sperm reservoir.     

Anatomy and Histology of the Male Reproductive Tract and Spermatogenesis Fine Structure in the Lesser Anteater (Tamandua tetradactyla,Myrmecophagidae, Xenarthra): Morphological Evidences of Reproductive Functions

The anatomy and histology of the male genital tract of the lesser anteater were studied. Fine details of spermatozoa regarding their genesis and morphology were also studied in six adult specimens. The testes lie in the pelvic cavity. The deferent duct emerges from the epididymis and opens into the ejaculatory duct, which drains into the membranous urethra. Accessory glands (prostate, seminal vesicle and bulbourethral gland) are histologically similar to those described in other mammals. The short penis presents an urethral orifice, while the corpus spongiosum becomes thinner at the end indicating the absence of a histologically defined glans. The seminiferous epithelium shows: (1) Sertoli cells with deep nuclear indentations, (2) spermatogonia with crusty‐like chromatin, (3) spermatocytes at different stages of maturation and (4) three morphologically distinct stages of spermatid differentiation according to nuclear shape, acrosome development and chromatin condensation. Sperm heads appear oval. The length of the spermatozoa averages 67.33 ± 1.60 μm. Two specimens with inactive spermatogenesis were azoospermic. Their testes and epididymis presented sizes smaller than those with active spermatogenesis. These studies together with others in anteaters may contribute to successful breeding in conservation programmes.     

Influence of Boar Seminal Plasma on the Distribution of Spermatozoa in the Genital Tract of Gilts

The main purpose of the present study was to investigate whether boar seminal plasma affects the transport of spermatozoa in the genital tract of oestrous pigs or not, with special reference to the sperm transport into the oviducts. Altogether 17 gilts were used in three experiments.Experiment I. In nine gilts one uterine horn was injected surgically with 10¹⁰ spermatozoa suspended in seminal plasma and the other uterine horn with 10¹⁰ spermatozoa suspended in TESNaK-glucose buffer solution. The sperm deposition was performed under general anaesthesia. The gilts were slaughtered 1–2 or 4–6 h after insemination. The genital tract was removed and the numbers of spermatozoa determined in oviducts and in uterine horns.Experiment II. The insemination doses were prepared exactly as in Experiment I. Approx. 24 h before insemination Polyvinylchloride cannulas were inserted into the uterine lumen of the horns, drawn via the midventral incision at linea alba subcutaneously to cutaneous incisions ventral to the vulva opening. One cannula was placed in each uterine horn. At standing heat the insemination doses were slowly injected through the cannulas. The gilts were slaughtered 1 h after insemination and the numbers of spermatozoa within the genital tract were counted.Experiment III. In three gilts under general anaesthesia the uterine horns were ligated 10 cm from the uterotubal junction. The semen doses (containing 2 × 10⁹ spermatozoa), prepared as in Experiment I, were deposited into the uterine horns anterior to the ligatures through a cannula. The gilts were slaughtered 1 h after insemination, and the numbers of spermatozoa within the oviducts and ligated part of the uterine horns were counted.In all three experiments more spermatozoa were, on average, recovered in the oviducts connected to uterine horns inseminated with spermatozoa suspended in seminal plasma. In Experiments I andII this was the case for 10 of 14 gilts and in Experiment III for all the three gilts. It is therefore suggested that boar seminal plasma pro¬motes sperm transport into the oviduct of oestrous pigs. The back¬ground mechanism for this is discussed.     

不同季节肉用种公牛精液品质与生理常值、血清及精清生化指标相关关系研究

以利木赞和夏洛来种公牛为试验牛，研究不同季节对其精液品质、冻精产量、生理常值、血清及精清生化指标的影响。结果表明：（1）与其它季节相比，夏季肉用种公牛的原精活力、冻精活力、活精于百分数、顶体完整率均显著降低，而精于畸形率显著升高（P〈0．05）。精子密度以秋季最低。（2）冻精产量夏季处于最低水平，平均每月仅为7388支。（3）夏季肉用种公牛的血清LH、睾酮、T3、皮质醇的含量分别为48．12、4．15、1．11、4．89ng／L，显著低于春、秋、冬季（P〈0．01）。（4）夏季肉用种公牛血清钠、钾、钙、镁均处于最低水平，显著低于其它季节（P〈0．01）。（5）夏季肉用种公牛精清睾酮水平明显下降，显著低于春季、秋季和冬季。（6）精清钠、钾、钙夏季含量最低，分别为83．12mmol／L、18，77mmol／L和4．63mmol／L；精清镁含量由春季至秋季呈显著下降趋势；精清磷含量夏季最高，为4．65mmol／L，显著高于春季、秋季和冬季。（7）肉用种公牛精清ALP夏季含量为1784．3U／L，显著高于春、秋两季，但低于冬季。（8）精清睾酮与精液品质各指标存在显著相关；血清生化指标与精清生化指标存在显著相关；直肠温度、呼吸频率与精液品质及血清生化指标存在显著相关性，与精清生化指标相关性不大。     

精浆和精子对母兔子宫免疫及孕向发育的影响

在结扎子宫颈的情况下,分别在兔同一子宫的两侧子宫角剖腹注入生理盐水(对照)、精浆或精子以研究精浆和精子对子宫免疫和孕向发育的影响。试验共用母兔18只,每组6只。在注入后72 h时处死试验用兔,取子宫进行一般测量,并切片染色,用图象采集分析系统进行研究。结果表明:1)注入子宫的精浆和精子,都能明显降低黏膜中的肥大细胞数量和增加子宫角中冲洗液内的细胞数(P0.01)。精浆与精子相比,精浆降低黏膜中肥大细胞数量的作用更大(P0.05);而精子提高子宫角冲洗液内细胞数的作用更强(P0.01)。2)无论精浆还是精子,都能显著(P0.05)或极显著(P0.01)促进子宫(子宫角指数、子宫角长度和子宫角直径)、子宫壁(子宫壁厚度、黏膜层厚度、黏膜上皮层厚度和肌层厚度)、黏膜腺体(腺体数和腺上皮的厚度)以及黏膜血管的孕向发育。精浆与精子相比,就子宫、子宫壁及黏膜血管的孕向发育来说,精浆的作用更明显(P0.05或P0.01);而对子宫黏膜腺体孕向发育的作用,精浆和精子间没有明显差异(P0.05)。     

The Effect of Removal of Seminal Plasma, Egg Yolk Level and Season on Sperm Freezability of Canary Buck (Capra hircus)

Goat semen is different from that of other domestic species in its limited tolerance to the inclusion of egg yolk in the freezing medium, and this tolerance depends on the presence of enzymes in the seminal plasma that react with egg yolk, producing toxic compounds to the spermatozoa. Moreover, the goat is a seasonal breeder that shows variations in semen quality throughout the year, and those variations may affect semen freezability; hence in freezing protocols, for instance, removal of seminal plasma (washing) yields varying results. This work was designed to study this problem in Canary goats: semen from six males was collected in spring, autumn or winter, washed or non-washed, diluted in a freezing extender with 1.5, 6 or 12% egg yolk, frozen, and thawed after 2 days, 2 or 6 months of cryopreservation. The effect of egg yolk concentration in the freezing extender was far more important than the effect of washing or season on sperm cryosurvival. The quality of frozen-thawed semen tended to improve as egg yolk concentration increased regardless of the effects of season, washing or period of cryopreservation. Washing produced a positive effect on frozen-thawed semen collected during spring or autumn, but the difference decreased as the concentration of yolk increased. However, washing produced a negative effect on frozen-thawed semen collected during winter, diluted with either 6 or 12% egg yolk. There was no apparent seasonal effect on gross measures of sperm production but the seasonal effect was ever present and was reinforced by freezing.     

Motility of Fresh and Frozen-Thawed Stallion Sperm from Three Segments of the Epididymal Cauda and the Effect of Post-Thaw Seminal Plasma Addition on Motility

Preservation of epididymal spermatozoa is an advantageous method to preserve genetic material of endangered species or valuable breeding animals after sudden death and injuries. Despite lower pregnancy rates, fertilization with epididymal sperm has been proven successful. Variable sperm quality after cryopreservation among individual stallions and the usually terminal chance to preserve epididymal sperm are opportunities for the development of a freezing procedure suitable for the majority of stallions. To evaluate the effect of the preservation procedure, we analyzed the sperm motion characteristics after every step of processing. In addition, we investigated the influence of seminal plasma on motility of frozen-thawed semen. We compared three segments of the cauda epididymidis and harvested spermatozoa by retrograde flushing (most caudal part) and mincing (more cranial segments) to augment the number of spermatozoa. During processing, there were differences in sperm motion characteristics depending on the segment of the cauda epididymidis. Distinct increases in motility due to different extenders and the effect of seminal plasma suggest that motion characteristics of raw and frozen-thawed spermatozoa are strongly influenced by microenvironment and must be interpreted with caution.     

高、低营养水平饲粮对妊娠环江香猪繁殖性能、体成分和血浆生化参数的影响

本试验旨在研究高、低营养水平饲粮对妊娠母猪繁殖性能和体成分的影响,并探讨其生化机制。选用首次妊娠的环江香猪48头,根据体重随机分为2组,每组8个重复(栏),每重复3头。配种后分别饲喂NRC饲粮[消化能(DE)为14.73 MJ/kg,粗蛋白质(CP)含量为13.11%]和地方猪饲粮(DE为12.24 M J/kg,CP含量为9.77%)。分别于配种后45、75和110 d称取母猪体重,每栏选取1头母猪,前腔静脉采血用于生化参数分析,放血处死后解剖,记录胎猪头数,称取胎猪个体重,并测定母体成分。结果表明:与地方猪饲粮组相比,NRC饲粮组母猪的繁殖性能和体成分指标差异均不显著(P0.05),妊娠45和75 d时血浆谷丙转氨酶以及妊娠75 d时谷草转氨酶活性均显著降低(P0.05),妊娠45 d时低密度脂蛋白-胆固醇浓度,妊娠75 d时碱性磷酸酶活性、高密度脂蛋白-胆固醇和总胆固醇浓度以及整个妊娠期甘油三酯浓度均显著升高(P0.05);在不同妊娠期,2种饲粮对各测定指标的影响有一定差异。由此可见,2种饲粮对环江香猪的繁殖性能和体成分均无显著影响;NRC饲粮在一定程度上改善了机体代谢,促进了生长发育,但同时增加了脂肪沉积。     

发酵中药渣对妊娠母猪繁殖性能、血浆生化参数和抗氧化指标的影响

本试验旨在研究发酵中药渣对妊娠母猪繁殖性能的影响,并通过测定血浆生化参数和抗氧化指标探讨其作用机制。试验选用3~5胎次、体况和预产期相近的长白×大白二元母猪75头,随机分为3组,每组25头,对照组饲喂基础饲粮,中药渣组和发酵中药渣组分别在基础饲粮中添加1.5 kg/t的中药渣和发酵中药渣。试验周期为整个妊娠期,统计返情、空怀和流产的母猪数,窝产仔数、产活仔数以及仔猪初生窝重和初生个体重。分别于母猪妊娠第45天、第75天和第114天(分娩当天),每组随机选取8头母猪,耳缘静脉采血,分离血浆,测定生化参数和抗氧化指标。结果表明:饲粮添加中药渣或发酵中药渣对母猪的繁殖性能无显著影响(P0.05)。与对照组相比,妊娠第114天时,发酵中药渣组的血浆总蛋白(TP)含量显著升高(P0.05),中药渣组和发酵中药渣组的血浆高密度脂蛋白-胆固醇(HDL-C)和低密度脂蛋白-胆固醇(LDL-C)含量显著升高(P0.05);妊娠第45天时,中药渣组和发酵中药渣组的血浆过氧化氢酶(CAT)活性显著升高(P0.05),同时血浆碱性磷酸酶(ALP)活性显著降低(P0.05)。综上所述,饲粮添加1.5 kg/t发酵中药渣不影响妊娠母猪的繁殖性能,但可在一定程度上改善机体代谢,增强抗氧化功能。     

OC4Influence of Seminal Plasma Added to Highly Diluted Semen on Sperm Motility of Young Boar Feed with Selenium and Vitamin E

High dilution rates have been documented as detrimental for boar spermatozoa, shortening their lifespan (Centurion et al. 2003, Biol Reprod 69: 640–646). Addition of seminal plasma (SP) to semen extenders, or selenium (Se) and vitamin E (VE) in diet of boars could increase motility of highly diluted spermatozoa (HDS). The aim of this work was to evaluate the effect of seminal plasma on sperm motility of HDS from boars feed with Se and VE. Sixteen 12 month-old boars were designed to one of four dietary treatments: (i) control, Se 0 ppm–VE 0 IU/kg; (ii) Se 0–VE 250; (iii) Se 0.5–VE 250 and (iv) Se 0.5–VE 0. Boars were treated for 8 weeks before semen collection. Sperm rich fractions from each boar were diluted to 5 × 10⁶ sperm/ml in PBS medium and incubated at 37°C with or without 10% SP. The measurements were done at 0, 2 or 5 h. Data were analyzed as a mixed model for a factorial design [2 (Se) × 2 (VE) × 2 (SP) × 3 (h)]. Percentage of sperm motility (PSM) increases significantly (p < 0.001) with addition of Se (81.3 ± 1.52), VE (81.0 ± 1.62) and SP (81.5 ± 1.57) vs control (73.4 ± 1.61). There was significant interaction Se × VE (p < 0.001) and Se × VE × SP (p < 0.05) in PSM. However, PSM was affected significantly by time (0 h 83.4 ± 1.92; 2 h 80.7 ± 1.92 and 5 h 67.9 ± 1.92; p < 0.001). There was significant interaction SP × Time (p < 0.05) in PSM. These results indicate that Se, VE and SP improve seminal viability. Addition of 10% of SP maintains PSM at least during 5 hours.     

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     

初乳对犊牛胃肠道、内分泌和代谢的影响

犊牛在出生后一段时期内消化道、代谢和内分泌的变化十分明显。消化初乳和常乳的能力需要胃肠道特殊的结构和功能。初乳的成分在哺乳期开始后发生变化。初乳的摄入对被动免疫和营养素的供给非常重要。此外,初乳中还含有对犊牛生长起重要作用的激素、生长因子、细胞分裂素、多胺和核苷。血清中的必需氨基酸的模式和谷氨酸盐/谷氨酸酯的比例也依赖于是否饲喂初乳和喂初乳的时间。初乳喂养的时间和数量对激素有相当大的影响。本文综述了有关初乳对胃肠道、内分泌和代谢作用的研究进展。     

公火鸡外周血浆促黄体激素、睾酮浓度的变化及其与精液品质相关性的研究

8只公火鸡自然光照下饲养,每隔10d收集一次血样和精样,连续5个月(6-10月份)。用放射免疫法测定外周血浆的促黄体激素及睾酮浓度,并用常规方法估测精液品质。结果表明,血浆LH浓度从6月上旬到10月下旬逐步降低,其与白昼长度及湿度呈显著的相关性(r分别为0.553和-0.586)。血浆睾酮浓度的变化趋势与LH相似,血浆睾酮浓度与LH浓度、白昼长度呈显著的相关性(r分别为0.747和0.953)。射精量的变化与白昼长度的变化一致,相关系数为0.655。本实验还表明,LH与射精量呈弱正相关,而与精子密度和死亡精子百分数呈中等强度正相关(r分别为0.368、0.591和0.553)。睾酮浓度与精液品质指标皆呈弱正相关。公火鸡换羽时,血浆LH浓度、射精量,精子密度及死亡精子百分数显著低于非换羽期,睾酮浓度没有发生明显的变化。     

Effect of Storage Time and Temperature of Equine Epididymis on the Viability,Motion Parameters,and Freezability of Epididymal Sperm

The recovery of sperm from the epididymal cauda may be the last chance to obtain genetic material when sudden death or serious injuries occur in valuable stallions. However, the lack of technical knowledge regarding the storage and transportation of the epididymis often prevents the preservation of the sperm. Therefore, the aim of this study was to compare sperm parameters of sperm obtained immediately after orchiectomy with sperm recovered from epididymal cauda at different times after storage at 5°C and at room temperature (RT). For that, 48 stallions of different breeds were used. In group 1 (control group), eight stallions were used, and the harvest of the epididymal sperm was performed immediately after orchiectomy. In group 2, 40 stallions were used, which were divided into five groups according to the storage time of the epididymis after orchiectomy (6, 12, 18, 24, or 30 hours), making a total of eight stallions per group. One epididymis of each stallion was stored at 5°C, and the contralateral epididymis was stored at RT, both for the same period. The sperm parameters of total motility, progressive motility, progressive linear velocity, curvilinear velocity, percentage of rapid sperm, and plasma membrane integrity were evaluated in all the groups after sperm recovery, resuspension in a sperm freezing diluent, and thawing. In conclusion, the storage of the testis-epididymis complex at 5°C provided better preservation of epididymal sperm than the storage at RT, and regardless of the temperature, the progressive motility is the sperm parameter that is most sensitive to storage time.     

不同蛋白质水平下种公猪精液品质与血液生化指标的相关性研究

通过对成年健康大约克公猪饲喂不同能量、蛋白质含量的饲料,研究不同能量、蛋白质含量条件下血液生化指标(包括内分泌激素和其他血液指标)与精液品质的相关性。结果表明:在本次试验中,能量对精液品质和血液生化指标的影响差异不显著(P>0.05);hTsh浓度与精子活率存在相关性且受蛋白质水平的影响;ALB浓度与精子密度的相关系数随着蛋白质水平的升高约有下降,但它与精子畸形率的相关系数却上升。