Factors Affecting Pregnancy Rate in Artificial Insemination with Frozen Semen During Non-Breeding Season in Murciano–Granadina Goats: a Field Assay

An artificial insemination programme was carried out to study the effect of factors such as depth of semen deposition, inseminator skill, farm, sire and expression of oestrus on pregnancy rate in Murciano-Granadina (MG) goats during non-breeding season and using frozen semen. Frozen-thawed semen from six males was applied by three technicians to inseminate a total of 551 goats in 17 farms distributed throughout the Mediterranean area of Spain. Pregnancy rate was determined at 6 weeks after insemination by transabdominal ecography. Overall pregnancy rate was 57%. Farm and depth of semen deposition affected pregnancy rate, whereas the sire and the technician had no effect. The deeper the semen was deposited in the genital tract, the higher was the rate of pregnancy obtained, being greater when the catheter reached the uterus. In spite of the relevant difference observed (48.2% vs 59.0%), pregnancy rate of females not coming into oestrus until 30 h after sponge removal was not significantly different, compared with those showing oestrus during the OD procedure. In conclusion, our field assay data on AI in MG goat with frozen-thawed semen showed that post-cervical insemination presented significantly greater pregnancy rate in comparison to when semen is deposited in the vagina or in the caudal part of the cervix.     

Effect of Artificial Insemination Protocol and Dose of Frozen/Thawed Stallion Semen on Pregnancy Results in Mares

Deep intra‐uterine insemination is commonly accepted as a routine procedure for artificial insemination in horses. The motives and principles of deep insemination are well described, but the equipment used may differ. In this trial, the efficiency of two different insemination pipettes for deep intra‐uterine insemination in the mare was compared with insemination into the uterine body using commercially available frozen–thawed semen of two stallions of proven fertility. These inseminations were performed using two different doses. The semi‐flexible Minitube pipette was compared with a newly designed insemination device with a more flexible telescopic insemination catheter (Ghent device). The semi‐flexible Minitube pipette performed better than the newly designed insemination device with respect to pregnancy outcome (p = 0.008). The superiority of deep horn insemination over uterine body insemination was reflected by the better pregnancy rates obtained after deep insemination using the same low doses (30.6% better pregnancy rates) (p = 0.0123).     

临沂市奶山羊鲜精人工授精技术应用

临沂市具有较好的农业资源,适合发展奶山羊养殖业。但是,目前奶山羊传统的自然交配繁育速率较慢,且受胎率低,不利于奶山羊养殖业的快速发展。近年来,由于人工授精技术具有增强繁殖速率、提升受胎率、提高种公羊利用率和降低成本等优点,在畜牧养殖业中的运用和推广十分广泛。但是人工授精的成功率受到母羊发情期、精液采集和保存、输精方式等多种因素的影响,因此增强对该技术的了解,对促进当地奶山羊的良种繁育具有重要意义。     

如何做好鸡的稀释精液人工授精

良好的饲养管理工作是稀释精液人工授精技术的基础。母鸡日粮中的能量、蛋白质要符合产蛋需要，特别是钙磷含量和比例要适当，维生素和微量元素不能缺乏，否则，即使是产蛋率上去了，受精率、孵化率和健雏率也不会高。另外，鸡舍的环境卫生要     

荷斯坦奶牛性控冻精人工授精应用效果的研究进展

近年来,对性别控制的研究呈现出迅速发展的趋势。应用性控冻精进行人工授精,是快速增加优质高产奶牛数量的有效途径。奶牛性控冻精的人工授精技术是将奶牛种公牛的精液通过精子分离仪使含X、Y染色体精子得到有效地分离,再将分离后得到的X精子冷冻制成冻精,进行人工授精,从而使母牛怀孕产母犊的技术。     

Development of Procedures for Sex-sorting Frozen–Thawed Bovine Spermatozoa

Dairy bull sperm may be sex‐sorted, frozen and used to artificially inseminate heifers with acceptable fertility if the herd is well‐managed. One drawback to the technology is that donor bulls must be located within a short distance of the sorting facility in order to collect semen, which limits the number of bulls from which sorted sperm are available. A successful method used to overcome this limitation in sheep is sex‐sorting from frozen–thawed semen and refreezing for artificial insemination. This technique is attractive to the dairy industry, and therefore a series of three experiments was designed to investigate the optimal methods to prepare, sex‐sort and re‐freeze frozen–thawed bovine sperm. Sperm were prepared for sorting by density gradient separation in either PureSperm^® or BoviPure?, followed by staining in one of three diluents (Androhep^®, Bovine Sheath Fluid + 0.3% BSA or TALP buffer). Sperm were sorted and collected into Test yolk buffer, and frozen in an extender containing 0, 0.25, 0.375 or 0.5% Equex STM Paste. Frozen–thawed sperm were better orientated (p = 0.006) and had fewer damaged membranes (8.7 ± 0.6% vs 19.5 ± 2.4%; p = 0.003) after centrifugation in PureSperm^® rather than BoviPure? gradients. Sperm orientation (p < 0.05) and motility (69.9 ± 3.0 vs 55.6 ± 4.0; p < 0.001) were highest after staining in Androhep^® rather than in TALP buffer. Sperm were more motile (58.2 ± 4.7 vs 38.7 ± 3.5; p < 0.001) and had better acrosome integrity (74.3 ± 2.9 vs 66.8 ± 2.0; p < 0.001) after freezing in an extender containing 0.375% Equex STM Paste than in extender without Equex. Hence, a protocol has been developed to allow frozen–thawed bull sperm to be sex‐sorted with high resolution between the sexes, then re‐frozen and thawed with retention of motility and acrosome integrity.     

天祝白牦牛细管冻精授配试验报告

[目的]为加快天祝白牦牛保种选育的进程,提高天祝白牦牛优秀种牛的利用率,[方法]利用天祝白牦牛细管冻精制作成功的前提下,开展天祝白牦牛人工授精试验.[结果]授配285头,受胎193头,受胎率为67.7%,产犊成活183头,死亡10头,成活率94.8%,[结论] 说明野外人工冻精授配技术应用于天祝白牦牛保种选育工作是可行性的.     

蓝狐人工授精及精液冷冻技术研究与应用

以芬兰种公狐为父本,本地饲养的北极狐为母本进行杂交改良。通过人工授精的方法,提高种公狐的利用率。采用精液冷冻技术,可解决公母狐因发情的时间差而导致的空怀,从而提高受配率、准胎率,最终提高蓝狐养殖业的经济效益。     

弯头输精针输配山羊效果的观察

近年来，随着山羊人工授精技术的大力推广，为市场提供了大量的优质肉羊，使养殖农户增收了可观的经济效益。据调查，羊改点输精员使用直头金属输精针，输配山羊往往在进针时较为困难，造成了受胎率不高，为解决输精员进针技术难题，根据山羊生殖器官形态特征，我们采用了自己设计制作弯头输精针输配山羊取得了较好效果，现介绍供同行参考。１材料与方法１．１材料（１）设计制作５０支弯头金属输精针（长１９cm，针体直径２mm、从套注射器端起至针体１７．５cm处处理形成３０度的曲弯头，曲弯头长度１．５cm，按提供尺寸经江苏省泰兴市动物…     

Birth of Kids After Artificial Insemination with Sex‐Sorted,Frozen‐Thawed Goat Spermatozoa

Successful sex‐sorting of goat spermatozoa and subsequent birth of pre‐sexed kids have yet to be reported. As such, a series of experiments were conducted to develop protocols for sperm‐sorting (using a modified flow cytometer, MoFlo SX^®) and cryopreservation of goat spermatozoa. Saanen goat spermatozoa (n = 2 males) were (i) collected into Salamon's or Tris catch media post‐sorting and (ii) frozen in Tris–citrate–glucose media supplemented with 5, 10 or 20% egg yolk in (iii) 0.25 ml pellets on dry ice or 0.25 ml straws in a controlled‐rate freezer. Post‐sort and post‐thaw sperm quality were assessed by motility (CASA), viability and acrosome integrity (PI/FITC‐PNA). Sex‐sorted goat spermatozoa frozen in pellets displayed significantly higher post‐thaw motility and viability than spermatozoa frozen in straws. Catch media and differing egg yolk concentration had no effect on the sperm parameters tested. The in vitro and in vivo fertility of sex‐sorted goat spermatozoa produced with this optimum protocol were then tested by means of a heterologous ova binding assay and intrauterine artificial insemination of Saanen goat does, respectively. Sex‐sorted goat spermatozoa bound to sheep ova zona pellucidae in similar numbers (p > 0.05) to non‐sorted goat spermatozoa, non‐sorted ram spermatozoa and sex‐sorted ram spermatozoa. Following intrauterine artificial insemination with sex‐sorted spermatozoa, 38% (5/13) of does kidded with 83% (3/5) of kids being of the expected sex. Does inseminated with non‐sorted spermatozoa achieved a 50% (3/6) kidding rate and a sex ratio of 3 : 1 (F : M). This study demonstrates for the first time that goat spermatozoa can be sex‐sorted by flow cytometry, successfully frozen and used to produce pre‐sexed kids.     

欧拉型藏羊的人工采精与细管冷冻精液授配技术研究

为提高青海省本地优质藏羊选育工作,笔者对青海省果洛州玛多县6只欧拉型藏公羊和20只欧拉型藏母羊进行优质选育试验,结果表明:藏公羊鲜精采集量平均为1.5mL,呈乳白或乳黄色、无气味、有云雾状,冻前鲜精活力平均为0.65,平均活力高出国家羊精子标准活力的0.075,冻后活力为0.3到0.5,平均活力高出国家羊精子标准活力0.1。且经冷配后的20只藏母羊均无返情现象。本试验对于藏羊的改良有重要的实践意义,对于改善纯藏羊品种也有重要的参考指导意义。     

奶牛性控冻精人工授精技术的推广效果调查研究

通过对性控冻精人工授精技术推广,并将其推广效果作了调查研究,结果利用性控冻精进行人工授精,可大幅度提高奶牛优良母畜的比例,加快牛群品质改良的进程。通过技术推广3年共受配奶牛254头,怀孕161头,受胎率平均63.8%,生产牛犊130头,生产母犊120头,生母率平均89.3%,该项技术在奶牛生产中有很大的应用空间,可提高奶牛养殖业经济效益。     

Pregnancy Rate Obtained with Short-term Protocol for Timed Artificial Insemination in Goats

Recent studies demonstrated that the Short-term Protocol of 5 days of progestogen treatment plus one dose of prostaglandin F(2alpha) and equine chorionic gonadotrophin (eCG) resulted in a close synchronized ovulation (60 h after the end of treatment approximately). In addition, oestradiol benzoate (ODB) is effective in synchronizing ovulation in goats and could be an alternative to eCG. This study was performed to determine the pregnancy rate using the Short-term Protocol comparing: (i) two different moments of timed artificial insemination (TAI) after eCG treatment (trial 1) and (ii) ODB as an alternative to eCG treatment (trial 2). In trial 1, 250 IU of eCG was given at the end of progestogen exposure, and cervical TAI with fresh semen was performed 48 h (n = 156) or 54 h (n = 168) later. In trial 2, 250 IU of eCG was given at sponge withdrawal (eCG group, n = 154) or 200 mug of ODB was given 24 h later (ODB group, n = 119). TAI was performed 54 h after the end of progestogen treatment. Pregnancy rate was determined by transrectal ultrasonography. In trial 1, the pregnancy rate for goats with TAI performed at 54 h (107/168, 63.7%) was higher than for those with TAI performed 48 h (77/156, 49.4%; p < 0.05) after sponge withdrawal. In trial 2, pregnancy rate was higher in eCG (94/154, 61.0%) than in ODB (49/119, 40.3%; p < 0.05) treated goats. In conclusion, the highest pregnancy rate was achieved using Short-term Protocol associated with eCG and TAI performed 54 h after treatment.     

Effect of Cloprostenol Treatment at Artificial Insemination on Sow Fertility

In two experiments, we examined the effect of administration of cloprostenol at artificial insemination on fertility of multiparous sows. Lactation length was 21 days and only sows exhibiting estrus by 7 days after weaning were included. Cloprostenol was administered by IM injection (175 microg; n=126) or supplemented in the semen dose (525 microg; n=129). Control sows (n=130) received no hormone. In either experiment, there was no effect of any treatment on subsequent litter sizes. In experiment 1, there was no effect of treatment on farrowing rate, but in experiment 2, farrowing rate was higher (p < 0.05) in sows receiving cloprostenol by injection. Taken together, these results indicate no consistent effect of cloprostenol administration by intramuscular injection or by inclusion in the semen dose on sow fertility and so its routine use is not warranted.     

不同胎次奶牛应用性控冻精的效果观察

本文探讨了不同胎次奶牛应用性控冻精人工授精的效果。观察了不同胎次奶牛使用性控冻精后的情期受胎率、流产率、正产胎儿成活率、产母犊率、产公犊率和正产胎儿死亡率。结果表明,情期受胎率、正产胎儿成活率和母犊率随着胎次的增加而呈下降趋势（P〈0．05）,而流产率、产公犊率和正产胎儿死亡率随着胎次增高而逐渐增加（前一项P〈0．05,后两项P〈0．01）。说明不同胎次奶牛对性控冻精的应用效果具有明显的影响。     

奶牛冷冻性控精液的人工授精试验

XY与CRI奶牛性控冻精解冻后精子活力为0.38、0.41,密度为210万个/剂和230万个/剂,常规冻精解冻后精子活力为0.48,密度为1020万个/剂.据试验,室温下性控精液在5～15 min时其活力下降比常规冻精明显,奶牛发情结束前后4 h内的情期受胎率分别为63.3%与70.7%;青年牛2剂/次、1剂/次的受胎率分别为60.71%(17/28),52.63%(10/19),深度输精与宫体输精的受胎率分别为66.66%(18/27),57.14%(20/35).所产后代母犊率达95.91%(47/49).     

Artificial Insemination with Frozen Semen in Ewes at Different Times of the Breeding Season

Totally 13575 ewes of two different breeds, Dala and Spel, were inseminated with semen, frozen in straws and thawed at 70°C for 8 sec. An insemination dose of 0.2 ml containing approx. 150 × 10⁶ spermatozoa with at least 45 to 50% progressive motility was imerted 5 to 12 mm into the cervix. The insemination was performed once between 12 and 30 h after the onset of heat. The NR rate of the Dala ewes increased significantly during the season. The NR rate of the ewes inseminated before 15. November was 44.3%, from 15. to 20. November 52.2%. from 20. to 25. November 55.3% and from 25. November and later 61.4%. The corresponding values for the ewes of the Spel breed were 57.3, 58.7, 61.5 and 71.0% respectively, and only the difference between the two last values was statistically significant. The difference between the fertility of the two breeds was significant within each of the periods .     

Fertility in Dairy Cows After Artificial Insemination Using Sex‐Sorted Sperm or Conventional Semen

The aim of this study was to compare pregnancy per artificial insemination (P/AI) after timed AI with sex‐sorted sperm (SS) or conventional semen (CS) in lactating dairy cows. Cyclic cows (n = 302) were synchronized by Ovsynch and randomly assigned into two groups at the time of AI. Cows with a follicle size between 12 and 18 mm and clear vaginal discharge at the time of AI were inseminated with either frozen‐thawed SS (n = 148) or CS (n = 154) of the same bull. A shallow uterine insemination was performed into the uterine horn ipsilateral to the side of probable impending ovulation. Pregnancy per AI on Day 31 tended (p = 0.09) to be less for SS (31.8%) than CS (40.9%). Similarly, P/AI on Day 62 was less (p = 0.01) for cows inseminated with SS (25.7%) compared with CS (39.0%). The increased difference in fertility between treatments from Days 31 to 62 was caused by the greater (p = 0.02) pregnancy loss for cows receiving SS (19.2%) than CS (4.8%). Cow parity (p = 0.02) and season (p < 0.01) when AI was performed were additional factors affecting fertility. Primiparous cows had greater P/AI than multiparous cows both on Day 31 (41.7% vs 25.0% in SS and 53.0% vs 31.8% in CS groups) and on Day 62 (33.3% vs 20.5% in SS and 48.5% vs 31.8% in CS groups). During the hot season of the year, P/AI on Day 31 was reduced (p = 0.01) in the SS group (19.6%) when compared with the rates during the cool season (38.1%). In conclusion, sex‐sorted sperm produced lower fertility results compared to conventional semen even after using some selection criteria to select most fertile cows.     

稀释液中添加棉子糖对冻融后绵羊精液品质的影响

本试验目的是研究稀释液中添加不同浓度的棉子糖对冻融后绵羊精液品质的影响。选用10只健康乌珠穆沁种公羊,采集精液经检验合格在37 ℃下混匀,精液样品平均分成5份,用含有0、5、10、15、20 mmol/L的不同浓度棉子糖稀释液稀释。采用0.25 mL细管法冷冻,将冷冻后的细管精液置于37 ℃水浴锅中解冻15 s后进行相关指标评价及酶(SOD、GSH、MDA、GSH-Px和CAT)的测定。结果表明,当棉子糖的添加量为15 mmol/L时精子活率显著高于对照组(P＜0.05),畸形率显著低于对照组(P＜0.05),弯尾率显著高于对照组(P＜0.05),SOD活力显著提高(P＜0.05),MDA也显著低于对照组(P＜0.05)。     

Single-Layer Centrifugation Through Colloid Positively Modifies the Sperm Subpopulation Structure of Frozen–Thawed Stallion Spermatozoa

The present study attempted to select the subpopulation of stallion spermatozoa that best survived a conventional freezing and thawing procedure, using centrifugation of post-thawed semen samples through a single layer of a glycidoxypropyltrimethoxysilane-coated silica colloid with a species-specific formulation (Androcoll-E™). After freezing and thawing, four sperm subpopulations were identified, listed as FT1 to FT4. While subpopulations FT1 and FT2 were characterized by low sperm velocity, high velocities characterized the ones called FT3 and FT4. The single-layer centrifugation (SLC)-handled sperm sample was enriched in subpopulation FT3, reaching a proportion of 82.6% of the present spermatozoa, in contrast with the non-filtered control post-thawed semen, where this sperm subpopulation only accounted for 16.3% of the total. It is concluded that in the equine industry, the SLC is a practical, easy-to-perform approach to improve the quality of equine frozen–thawed semen samples.