冬夏季长白猪精液中细菌多样性与精子质量特性关联分析

为探究冬夏季长白猪精液中细菌多样性变化和精子质量特性之间的相关性,利用16S rRNA基因高通量测序技术得到冬夏季长白猪精液中微生物的特征序列(ASV),计算机辅助精子分析系统(CASA)分析冬夏季精子活力参数指标的变化.结果 显示:长白猪精液中细菌多样性在冬夏季之间差异显著(P＜0.05),门水平冬季优势菌群为厚壁菌...     

Post-thawing Sperm Quality in Chilean Purebred Stallions: Effect of Age and Seasonality

In this work, we investigated the influence of age and seasonality on sperm motility and DNA fragmentation in post-thawing semen from Chilean Purebred Stallions (CPS), a horse breed presenting the oldest genealogy record in South America with an interesting reproductive industry. Despite that semen from aged CPS is frozen all year round, there is a lack of studies characterizing the breed semen freezability in accordance with age and seasonality. Twenty fertile CPS were grouped into the young group, the middle group, and the aged group. Ten ejaculates from each stallion were obtained by using an artificial vagina during summer (December) and winter (July) and directly frozen. Subsequently, the frozen semen was thawed and analyzed by a computer-assisted semen analysis and flow cytometer assessing progressive motility, mean velocity, and DNA fragmentation spermatozoa. Kruskal–Wallis test and Pearson’s correlation were used to determine statistical differences among groups and correlation among variables (P ≤ .05). Both spermatozoa motility traits decreased progressively in accordance with age and seasonality, showing the lowest values in the aged group during winter and the highest values in the young group during summer. Deoxyribonucleic acid fragmentation increased significantly in accordance with age and seasonality being highest in the aged group during winter and lowest in the young group during summer. Post-thawing sperm quality showed a negative correlation with the age of the stallions and a positive correlation with the normal sperm morphology before freezing. These results allow the conclusion that age and seasonality are important factors that need to be considered during the selection of CPS for reproductive programs.     

高温对荷斯坦种公牛精液品质及精清生化指标的影响研究

以荷斯坦种公牛为试验牛研究了高温对其精液品质和精清生化指标的影响。结果表明:(1)高温可造成种公牛精液品质显著下降。使原精活力、精子密度、活精子百分数和顶体完整率分别比春季下降3·13%(P<0·05)、34·8%(P<0·01)、15·0%(P<0·01)和17·8%(P<0·01),精子畸形率比春季上升25·5%(P<0·01)。(2)夏季荷斯坦种公牛精清睾酮含量仅为4·31pg/mL,极显著低于非高温季节(P<0·01)。(3)高温环境使得荷斯坦种公牛精清钾、钙、镁处于最低水平(P<0·01),精清碱性磷酸酶(ALP)、酸性磷酸酶(ACP)处于最高水平(P<0·01),使精清钠由春季至夏季呈显著下降趋势。     

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。     

季节变化对不同品种种公牛精液品质及产能的影响

选择年龄在3~4.5岁的不同品种的种公牛20头,观察四季变化对其精液次均采集量、平均密度、平均活力、精液利用率及制冻数量的影响。结果显示,在四个季节中,鲁西牛春、夏季的精液密度极显著高于其他季节(P＜0.01),秋季精子活力极显著低于春季(P＜0.01),夏、秋季的精液利用率及制冻数量显著低于春季(P<0.05);蒙贝利亚牛在夏季的精液采集量显著高于其他季节(P＜0.05),夏、秋季精液密度均显著低于春季(P＜0.05,P＜0.01),夏季精子活力显著低于春季(P＜0.05),夏、秋两季精液利用率及制动数量显著低于春季(P＜0.05);德系西门塔尔牛夏季精液采集量有明显上升,显著高于冬季(P＜0.05),秋季的精液密度显著低于春季(P＜0.05),而夏、秋季精子活力显著低于春、冬季(P＜0.01,P＜0.05),春季的精液利用率及制冻数量显著高于冬季与夏、秋季(P＜0.05,P＜0.01)。综上所述,季节变化均会对各品种公牛精液指标产生不同程度的影响,其中对鲁西牛的影响较小。     

Seasonal Variations in Seminal Plasma Proteins of Buffalo

The study was designed to evaluate the influence of season on semen characteristics and seminal plasma protein profile of buffalo bull semen. Thirty‐six ejaculates were collected in three seasons (winter, summer and rainy) from six adult Bhadawari bulls, and semen characteristics were evaluated immediately after collection. The seminal plasma was harvested by centrifugation and protein profiling, and percentage protein fractions were analysed by SDS‐PAGE. The significant effect of season was observed on ejaculate volume, sperm concentration, progressive motility, percentage live spermatozoa, hypo‐osmotic swelling test (HOST) and acrosomal integrity. The electrophoretogram of seminal plasma proteins revealed 20 protein bands in winter, 23 bands in rainy and 25 bands in summer seasons, illustrating the significant effect of seasons on seminal plasma proteins. Among these protein bands, 18 bands were observed common in semen samples of all three seasons while protein bands of 46, 55, 58, 144 and 160 kDa were found in rainy and summer seasons. The protein bands of 48 and 60 kDa were observed only in winter season, whereas 184 and 200 kDa were reported in summer season only. The protein fractions (protein%) of common protein bands observed in three seasons revealed a significant effect of season on protein bands of 24.5, 66, 70, 72, 84 and 86 kDa. From the study, it was pertinent that bull seminal plasma contains specific proteins in particular season, which may be associated with some of the semen characteristics, and these proteins could be used as markers of the semen quality of buffalo bulls.     

奶牛人工授精受胎率影响因素和提高措施

奶牛人工授精是以人工方法,采用机械手段采集优质种公牛的精液,再经过机械手段将特定处理后的精液输入母牛生殖道的一种生产技术方式。人工授精操作能极大减少种公牛的养殖数量,节约养殖成本,提高优质种公牛的利用效率,加快品种改良进程,促进育种工作的开展。但由于奶牛人工受精是一个侵入性的操作,一旦操作不合理或者各个细节把握不完善,会造成繁殖母牛出现严重的繁殖障碍,降低母牛的受胎率,不利于提高品种改良和扩大养殖效益。该文主要论述奶牛人工受精受胎率的影响因素,提出相应的改进措施。     

The Effect of Seasonal Thermal Stress on Lipid Mobilisation,Antioxidant Status and Reproductive Performance in Dairy Cows

Heat stress is a major factor contributing to low fertility of dairy cows with a great economic impact in dairy industry. Heat‐stressed dairy cows usually have reduced nutrient intake, resulting in a higher degree of negative energy balance (NEB). The aim of this study was to investigate the seasonal thermal effect on lipid metabolism, antioxidant activity and reproductive performance in dairy cows. Thirty‐two healthy dairy heifers were included in the study. According to the ambient temperature, animals were divided into two groups: winter (N = 14) and summer season (N = 18). Metabolic parameters, paraoxonase‐1 (PON1) activity and total antioxidant status (TAS) were monitored at the time of insemination (basal values) and from 1 week before until 8 weeks after calving. Number of services per conception and calving‐to‐conception (CC) interval were calculated from the farm recording data. Serum triglyceride, non‐esterified fatty acids (NEFA) and beta‐hydroxybutyrate (BHB) concentrations were significantly increased after calving in summer compared to winter, indicating higher degree of NEB in cows during summer. PON1 activity was significantly decreased after calving in both summer and winter group. TAS concentration was significantly lower in summer than that in winter. A significantly higher number of services were needed for conception in summer compared to winter, and CC interval was significantly longer in summer than that in winter as well. Additionally, reproductive performance significantly correlated with the severity of NEB, suggesting that lipid mobilization and lower antioxidant status contributed to poor reproduction ability in dairy cows during hot months.     

The Effect of Season on Semen Characteristics and Freezability in Bos indicus and Bos taurus Bulls in the Southeastern Region of Brazil

The objectives of this study were to evaluate the effects of season in southeast of Brazil comparing genotypes on semen characteristics, freezability and peripheral plasma concentrations of testosterone. Ejaculates of five Bos indicus bulls and six Bos taurus bulls were evaluated over a period of 27 months, which was divided into winter (July, August, September), spring (October, November, December), summer (January, February, March) and autumn (April, May, June). Semen was evaluated according to standard procedures for ejaculate volume, sperm concentration, gross-motility, progressive motility and sperm morphology. After preparing and freezing the ejaculates according to commercial procedures, the straws were stored in liquid N₂ until post-thaw evaluation. Ejaculate volume, sperm concentration, gross-motility, progressive sperm motility, vigor and morphological sperm defects were significantly influenced by season and genotype (p < 0.05). Heat tolerance was better in B. indicus bulls than in B. taurus bulls characterized by lower values of sperm abnormalities throughout the observation period. The highest values were recorded for abnormal heads followed by cytoplasmatic droplets in B. taurus bulls. The proportion of ejaculates which were eliminated before freezing for reasons of bad quality was lower in the B. indicus bulls. Temporal changes in peripheral plasma testosterone concentrations were higher in B. indicus bulls than in B. taurus bulls not revealing seasonal influences. The results of this study show clear genotype differences regarding semen quality. Freezability of B. taurus semen varies considerably throughout the year, leading to a high proportion of eliminated ejaculates. Collecting semen from B. taurus bulls during the summer in an artificial insemination centre may not be profitable.     

Exploratory analysis of differences in sperm morphology in Nelore and Gir (Bos indicus) bulls

The aim of this study was to evaluate the effects of breed and season on semen quality parameters of zebu bulls. Data (1,632 registers) of semen production from Gir (n?=?4) and Nelore (n?=?15) bulls were collected between October 2005 and November 2009. The ejaculates were collected twice a week during various seasons (summer, fall, winter, and spring) and evaluated for the following semen parameters: ejaculate volume, sperm concentration, sperm motility, forward progressive motility (FPM), and sperm morphology. Factor analysis was used to determine the relationship among variables. The effect of breed (Gir and Nelore) and season and their cross effect on each parameter and extracted factor were tested using ANOVA. A negative correlation (P?<?0.05) was observed between FPM and proximal droplet, as well as with abnormal loose head, abnormal small head, pouch formation, abnormal mid-peace, and strongly folded tail. Gir bull sperm showed more major defects, detached acrosome, and minor FPM (P?<?0.01), whereas Nelore bulls showed a higher number of sperm with normally loose head.     

A preliminary study on using autologous and heterologous boar sperm supernatant from freezing processes as post-thawing solution: its effect on sperm motility

The aim of this study was to evaluate the effect of post-thawing dilution with autologous and heterologous sperm supernatant on motility of frozen-thawed boar spermatozoa. During the cryopreservation, sperm supernatant (a combination of seminal plasma and semen extender, 50% v/v) or seminal plasma from nine boars (Duroc, Large White, and Landrace; three in each) was collected by centrifugation and stored frozen until use as post-thawing solution. Sperm pellets were further processed and cryopreserved using control-rate freezer and was thawed at 50°C for 12 s. After thawing, frozen thawed semen samples were diluted with seminal plasma (group A), supernatant from Landrace (group B), supernatant from Large White (group C), supernatant from Duroc (group D), and Modena™ semen extender (group E). Post-thawing motility was evaluated using a phase-contrast microscope after thawing at 1, 10, 20, and 30 min. The present results show that at 1 min, a significantly higher percentage (P ≤ 0.001) of progressive motility was found in groups B (53.3%) and C (53.9%) than the other groups. At 10 min, the highest (P ≤ 0.001) progressive motility was found in groups B (65%) and C (61%). At 20 and 30 min, a significantly higher percentage (P ≤ 0.001) of progressive motility was found in groups B (58.9%), C (53.5%), and D (45.6%) than groups A (3.9%) and E (20.6%). It can be stated that supernatant from the freezing processes (consisting of seminal plasma and Modena™, 50% v/v) had a beneficial effect on post-thawing progressive motility of frozen boar semen.     

The Effect of Season on Spermatozoa Motility,Plasma Membrane and Acrosome Integrity in Fresh and Frozen–Thawed Semen from Xinong Saanen Bucks

The aim of this study was to evaluate whether the season of ejaculate collection influences seminal quality parameters of pre‐ and post‐freeze–thawing in Xinong Saanen bucks. Ejaculates were collected from eight bucks throughout the four seasons (spring, summer, autumn and winter) in a 12 months’ time period, identified in the Northern Hemisphere. Semen samples were evaluated by the combinations of conventional and Computer‐Assisted Sperm Analysis (CASA) when fresh and after frozen–thawed, respectively. The results clearly demonstrated that season of ejaculate collection influenced (p < 0.05) fresh semen quality. Highest semen quality was observed during autumn. On the contrary, undesirable indices (significantly lower, p < 0.05) were observed in winter as compared with the other remaining seasons. CASA has clearly shown the influences of seasonal variations on semen motility parameters. Furthermore, season of ejaculate collection was also found to influence sperm freezability. Semen characteristics after frozen–thawed followed a similar pattern with that of fresh ejaculate except in spring. The results revealed that sperm quality was higher (p < 0.01) in summer and autumn than in spring and winter. In conclusion, seasonal variation influences semen quality in Xinong Saanen bucks. In addition to summer and autumn, fresh ejaculates in spring can also be successfully used for AI. Sperm from ejaculates collected during summer and autumn are more suitable for cryopreservation. Hence, it is possible to increase the efficiency of goat breeding by manipulating the seasonal variations of semen quality for immediate AI and/or cryopreservation.     

Impact of selection for residual feed intake on breeding soundness and reproductive performance of bulls on pasture-based multisire mating

There is concern in the beef industry that selecting bulls for feed efficiency based on residual feed intake (RFI) may have a negative impact on bull reproductive performance and fertility. Here we investigated the impact of selection of bulls for low RFI on breeding soundness evaluation (BSE), reproductive performance, and fertility of bulls under natural service in multisire mating groups on pasture. Of the 412 RFI-tested bulls available, 98 (23.8%) were culled for performance, type, temperament, or other reasons, and 88 (21.4%) were culled for failing BSE, for an overall cull rate of 45.1%. From among the 314 bulls subjected to BSE, 32 (10.2%), 20 (6.4%), and 36 (11.4%) were culled for poor feet and legs, scrotal circumference, and semen quality, respectively. The BSE traits were not different (P > 0.10) between bulls categorized as either inefficient (+RFI) or efficient (-RFI), but the proportion of bulls that failed to meet the 60% minimum sperm motility requirement tended (P = 0.07) to be greater in the -RFI group than in the +RFI group (10.2% vs. 4.4%, respectively). In a subpopulation of 115 bulls, individual progressive sperm motility was greater (P < 0.05) in +RFI (85%) than -RFI (80%) bulls. A multisire natural mating experiment was conducted during 2 consecutive breeding seasons (2006 to 2007 and 2007 to 2008) using 18 +RFI and 18 -RFI bulls. The overall calving rate (calves born/cows exposed) was 72.9%. Mean number of progeny per sire was significantly greater (P < 0.01) in -RFI bulls (18.3) than in +RFI bulls (11.8). Selection for feed efficiency based on RFI appears to have no detrimental impact on reproductive performance and fertility in beef bulls bred in multisire groups on pasture. However, the decreased sperm motility and the greater number of progeny per sire associated with -RFI status need further investigation.     

Semen alterations in porcine rubulavirus-infected boars are related to viral excretion and have implications for artificial insemination

Porcine rubulavirus (PoRV), also known as blue eye disease (BED) of swine, causes respiratory and reproductive problems in pigs at several developmental stages. To study the effect of PoRV infection on semen production, five boars were infected with 1 x 10(6) TCID(50)/ml of PoRV strain PAC-3 and evaluated for 59 days post inoculation (DPI). Infected boars developed reproductive tract pathology that included swelling of the testes and epididymides. Analysis of the semen showed that the infection had little effect on semen production in four animals, but semen from one boar showed severe alterations in sperm concentration, motility, and morphology. When motility was analyzed in BTS-diluted semen after 24, 48, or 72 h, alterations were detected in all boars. Furthermore, viral antigen was detected in semen, the seminal plasma fraction, or sperm fraction from all boars. These results showed that PoRV is excreted via semen and, therefore, artificial insemination is a potential route of dissemination.     

Relationships among age, body weight, scrotal circumference, semen quality and peripheral testosterone and estradiol concentrations in pubertal and postpubertal Holstein bulls

In the present study, the correlations among age, body weight, scrotal circumference (SC), semen quality and peripheral testosterone and estradiol-17beta (E(2)) concentrations were investigated in pubertal (n=5) and postpubertal (n=7) groups of Holstein bulls over a 6 week period. There were significant positive correlations (P<0.01) among age, body weight and SC in both groups, and similar significant correlations between sperm motility and SC in pubertal bulls (P<0.01) and between sperm concentration and SC in postpubertal bulls (P<0.05). The sperm motility after collection (P<0.05) and after freezing and thawing (P<0.01) of the postpubertal bulls correlated positively with the E(2) concentration. Estrogen may be important for the function of postpubertal bull testes, in which it may regulate spermatozoa motility in vivo.     

Testicular development, epididymal sperm reserves and seminal quality in two-year-old Hereford and Angus bulls: effects of two levels of dietary energy

The effect of high (HED) and medium energy diets (MED), fed to Hereford (H) and Angus (A) bulls from 6 through 24 mo of age, on scrotal circumference (SC), paired testes weight (PTW), epididymal sperm reserves (ESR) and seminal traits were examined. Over 3 yr, 120 bulls were involved. Angus exceeded H for both SC and PTW. Hereford bulls in yr 2 had smaller SC than in yr 1 or 3 but the response for A was consistent. Year affected PTW. In yr 2 Hereford bulls fed HED had 75% fewer ESR than MED-H bulls (9.3 vs 37.2 X 10(9]. Comparably treated A bulls had similar ESR numbers (29.2 vs 33.4 X 10(9]. In yr 3, epididymal sperm reserves of HED-H were depressed by 35% compared with MED-H (23.1 vs 35.7 X 10(9], whereas HED-A had 14% fewer ESR than did MED-A bulls (28.6 vs 33.1 X 10(9]. It was not obvious why H bulls were more susceptible to the effects of HED. Seminal quality of HED bulls was inferior to that of MED bulls, particularly with respect to progressive motility and the incidence of sperm in which a crater defect of the head was present at 2 yr of age. In yr 2 all seminal traits were severely depressed in 2-yr-old HED-H. Feeding HED to young H and A bulls reduced their reproductive potential.     

Seasonal variation in semen quality of bulls and correlations with metabolic and endocrine parameters

Sperm output and semen quality of 17 bulls sampled over 12 months showed minimal output in mid-winter and late summer and minimum quality in late summer. Monthly measurements of luteinising hormone and testosterone concentration in plasma and testosterone concentration in semen were made over 12 months. Serum non-esterified fatty acids (NEFA), albumin and total protein were also measured for the final seven months of this period. Plasma testosterone showed a strong negative correlation with sperm numbers two months hence but not in the current month. Plasma testosterone by bull and semen testosterone by month was also correlated with sperm output. Plasma luteinising hormone three months and serum total protein two months prior was positively correlated with sperm numbers ejaculated and the normality of sperm morphology, possibly by affecting the luteinising hormone dependent A0 to A1 spermatogonial division. NEFA was correlated with initial and post freezing/thawing motility in the current month, possibly by affecting membrane stability. The value of examining the bull in diagnosing infertility of cows where nutritional stress may have occurred is suggested, as is the use of albumin/total protein and NEFA measurements as a prognostic aid for time to return to normality of function of such bulls.     

长白公猪血清和精浆中元素含量对精液品质的影响

本试验旨在研究不同精液品质特征长白公猪血清和精浆中元素含量的差异,分析血清和精浆中元素含量对精液品质的影响。基于107头长白公猪的1402次精液品质记录,将公猪群按照精液可利用率划分为3组:低利用率组(利用率<60%,n=21)、中等利用率组(60%≤利用率≤80%,n=27)和高利用率组(利用率>80%,n=59)。采集每头长白公猪的血清和精浆样品,采用电感耦合等离子体质谱法检测血清和精浆中营养元素和毒性元素含量。结果表明:1)低利用率组长白公猪的有效精子数和精子活力显著低于中等利用率和高利用率组(P<0.01),精子畸形率显著高于中等利用率和高利用率组(P<0.01)。2)不同利用率组间长白公猪的血清和精浆元素含量无显著差异(P>0.05);但在血清和精浆元素含量与精液品质参数相关性分析中发现,精浆铅元素含量与精子活力呈显著负相关(P<0.05),与精子畸形率呈显著正相关(P<0.05)。3)对精浆铅含量分组进一步分析发现,精浆铅含量为0μg/L时长白公猪的精子畸形率显著低于精浆铅含量>10.0μg/L时(P<0.05),精子畸形率降低约6.11%。总的来说,长白公猪精浆中毒性元素铅的存在会通过损害精子活力和形态,影响公猪精液品质。     

Effect of pre-freeze addition of cysteine hydrochloride and reduced glutathione in semen of crossbred Jersey bulls on sperm parameters and conception rates

This study was aimed to assess the effect of pre-freeze addition of cysteine hydrochloride and glutathione (GSH) on post-thaw sperm functional parameters and field fertility. The experimental bulls (n = 6) aged 4-6 years were used for this study. A total of 36 ejaculates (six ejaculates per bull) were collected and divided into three groups, group I (control), group II (5 mm cysteine hydrochloride) and group III (5 mm GSH). The extended semen samples with and without additives were filled in mini straw using automatic filling and sealing machine and cryopreserved in liquid nitrogen. Post-freeze seminal traits were recorded after thawing at 37°C for 30 s. The curvilinear velocity (VCL) and amplitude of lateral head displacement values were significantly (p < 0.05) higher in GSH than the cysteine and control groups. Post-thaw plasmalemma integrity was significantly (p < 0.01) higher in GSH group when compared to cysteine and control groups. The loss of acrosomal integrity was significantly (p < 0.01) lower in GSH than in cysteine and control groups. Addition of GSH non-significantly (p = 0.1) improved mitochondrial membrane potential (MMP) (per cent) than control group. This study indicates that the addition of cysteine and GSH suggestive (p = 0.08) of reducing lipid peroxide levels. The conception rate (%) in glutathione group was significantly (p < 0.05) higher than that in cysteine (68 vs 58) and control (49) groups. The post-thaw sperm progressive forward motility (r = 0.4) had moderate, but no significant correlation with conception rate. However, post-thaw straight-line velocity (r = 0.7), loss of acrosomal integrity (r = -0.8) and MMP (r = 0.9) had significant (p < 0.05) correlation with field fertility. This study indicates that the use of glutathione as semen additive may be used for improving post-thaw semen quality and overall augmentation of pregnancy in cows.