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动物精子的性别可通过流式精子分选仪和DNA标识进行鉴定,而利用鉴定性别的精子(即性控精子)并借助人工授精技术或其它授精技术产生的后代,在过去5年中估计已多达30000个(其中大多数是母牛)。有关文献资料证明,能有效地区分X精子和Y精子的唯一标记物是精子染色体中的DNA。众所周知,目前世界各地采用的方法是Beltsville精子分选技术,该技术根据X精子群和Y精子群中DNA相对含量上的差异,用荧光染液(Hoechst33342染液)标记精子,随后利用流式细胞仪分选经荧光标记的精子,从而达到分离精子的目的。目前,X精子或Y精子正常的生产速度是每小时1500万个,该项技术已在家畜、实验动物和动物园动物中应用,如果将该技术应用在人上,在预选后代性别比例上可达90%～95%的成功率。因动物品种不同,性控精子在动物体内的授精部位也不相同。常规的人工授精技术、宫内授精技术、输卵管内授精技术、用于胚胎移植的体外受精(in-vitro fertilization,IVF)技术或子宫角深部授精技术均能有效地使动物怀孕,至于利用哪种技术进行性控精子的精则取决于动物品种。尽管所有动物都能获得高纯度的分选精子,但是在实际生产中利用低剂量精子还难以让母猪怀孕。子宫角深部授精技术每次授精0.5～1.00亿个性控精子已能产生可喜的效果:利用特制的输精管,输入常规人工授精所需精子量的五十分之一的性控精子,足以使动物怀孕。性控精子通过常规的授精技术能够被猪成功利用前,还需重新设计输精管,同时输精的次数和每次授精时精子数量也需作进一步的研究。分选精子的低温保存技术已被牛人工授精普遍采用。尽管已能利用冷冻的性别分选精子生产小猪,但是性别分选精子经冷冻和解冻处理后在常规生产中的应用还未达到最理想的效果。本文将讨论猪精子性别分选的最新研究成果及其发展趋势,并重点探讨将性别分选技术应用于养猪生产中必须对其进行必要的技术开发。 相似文献
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就奶牛生产而言,如能将产母犊率提高10％。15％,则对迅速发展我国乳牛业能起很大作用。流式细胞仪精子分离法是根据X、Y两类精子在DNA含量上的差异来分离精子。分离X、Y精子并用于人工授精或显微授精是控制家畜性别最简单、可行的方法之一,它可在授精之前就控制其性别,避免了胚胎的浪费。现在,超数排卵利用性控精液生产性控胚胎是性控胚胎移植的主要胚胎来源。对优良种母牛进行超排生产性控胚胎,是充分挖掘良种母牛的遗传潜力和向社会提供大量优质性控胚胎的重要手段。本研究旨在检测和验证流式细胞分离仪分离精子的过程或染料对奶牛超数排卵后卵母细胞受精及胚胎质量的影响,以及通过精子分离后奶牛精液对受胎率及性别比率的控制。 相似文献
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利用流式细胞仪分离奶牛精液的性别控制效果观察 总被引：10，自引：0，他引：10
目前,常见的牛性别控制方法有早期胚胎性别鉴定法和利用流式细胞仪进行X、Y精子分离法两种。流式细胞仪精子分离法是根据X、Y精子在DNA含量上的差异来分离精子。分离后的X、Y精子用于人工授精或显微授精可在授精之前就控制其性别,因而被认为是更具有实用价值的方法。此次研究通过对分离后精子的人工授精效果和性别控制效果进行试验观察,以期为该方法的进一步应用提供依据。1材料与方法1.1公牛及采精公牛为大庆田丰生物有限公司饲养的荷斯坦牛,共12头。饲以全价营养日粮,每天喂给鸡蛋1枚。采用常规的假阴道法采集精液。1.2精液的处理将采… 相似文献
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流式细胞仪分离精子研究进展 总被引：3，自引：0，他引：3
本文概述了近年来用流式细胞仪分离动物X、Y精子的研究进展、存在的问题和应用前景。流式细胞仪分离精子的研究已经取得了很大进展 ,在某些动物 (如牛 )分离的精子已开始用于人工授精。尽管该技术还存在许多亟待解决的问题 ,但随着流式细胞仪的改进和与各项辅助生殖技术 (如手术授精、体外受精、胞质内精子注射、低剂量精子人工授精 )结合 ,性别分离精子必将在实践中得到广泛的应用 ,产生巨大的社会和经济效益 相似文献
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采用流式细胞分离仪分离的梅花鹿X和Y型冷冻精液与常规冻精对62头3组同期化处理的马鹿进行直肠把握人工授精。结果表明,0.25 mL/支含106个有效精子的X和Y型冻精产仔率分别为43%和37%,而0.25 mL/支含107个有效精子的常规冻精产仔率为55%,X、Y型冻精与常规冻精组间差异显著(P<0.05),所产后代性别比率分别为0∶10,9∶0和5∶6,X、Y型冻精与常规冻精组间差异显著(P<0.05),X、Y型冻精与常规冻精所产后代出生及60 d时的体重差异不显著(P>0.05)。 相似文献
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养猪业要想获得高的仔猪繁殖成活率,必须抓养猪产仔、成活和增重率的提高。现介绍几项技术措施,供参考。一、提高母猪产仔率1.抓好母猪的膘分母猪在妊娠期间保持八成膘。多饲喂青饲料,精料不能多加,总能量保持在5200大卡左右,日粮精料不超过1公斤,饲养水平配种期和怀孕初期高,怀孕中期低,后期高。 相似文献
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Román Espinosa-Cervantes Alejandro Córdova-Izquierdo 《Tropical animal health and production》2012,45(1):1-8
The ability to preselect or predetermine the sex of offspring prior to conception is a highly desired technological tool for assisted female breeding programs specifically for milk production, and in males, for meat production and increasing livestock numbers. The current technology is based on the well-known differences in X- and Y-sperm in the amount of DNA. The technology uses modified flow cytometric instrumentation for sorting X- and Y-bearing sperm. The method can be validated on the basis of live births, laboratory reanalysis of sorted sperm for DNA content, and embryo biopsy for sex determination. Currently, the sex of animals has been predetermined with 90 % accuracy by sexing spermatozoa. In the bovine breeding industry, flow cytometric sperm sexing has not fulfilled its original promise. Sexed sperm doses are too expensive for widespread application while the fertility of sexed sperm doses is lower than unsexed ones. Essentially all bovine sexed semen is frozen and then applied through artificial insemination (AI) or in vitro fertilization. There is still a need in the animal breeding industry to develop a technique for sperm sexing that provides sufficient spermatozoa for AI doses, does not compromise sperm fertility, and is widely applicable to a range of species. In this review, we will summarize the current state-of-the-art in sex preselection in domestic animals and some wildlife species using flow cytometric sperm-sorting of X from Y sperm based on DNA differences. 相似文献
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Pregnancy Rates in Lactating Holstein–Friesian Cows after Artificial Insemination with Sexed Sperm 总被引：1，自引：0，他引：1
The effects of artificial insemination (AI) using sexed sperm on pregnancy rates have seldom been studied in lactating dairy cows on commercial dairy farms. We evaluated pregnancy results after AI of 306 lactating dairy cows, of which 157 were inseminated with 2x10(6) frozen/thawed sexed sperm and 149 with 15x10(6) frozen/thawed unsexed sperm. The average pregnancy and calving rates were 21.0% and 20% for the sexed-sperm AIs and 46% and 45% for the unseparated control-sperm AIs respectively (p<0.001). The proportion of female calves derived from sexed-sperm AI was 82% compared with 49% for control AI (p<0.01). The proportion of live and healthy calves in single births was 100% for sexed-sperm AI and 97% for control AI (p>0.05). Our results indicate that AI with low-dose sexed sperm under field conditions in commercial dairy herds without oestrus synchronization results in significantly reduced pregnancy rates compared with normal-dose AI. Improved insemination strategies combined with increased sperm doses are needed before the use of sexed sperm can be of any significant benefit for the dairy and beef industry. 相似文献
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The Beltsville sperm sexing technology is currently the only effective means of altering the sex ratio of offspring in livestock. The method is based on the flow-cytometric separation of X- and Y-chromosome-bearing sperm based on X/Y DNA content difference. It is an effective means of producing progeny of predetermined sex in cattle, swine, sheep, and laboratory animals. The method involves treating sperm with a DNA-binding fluorochrome, Hoechst 33342, and flow-cytometrically sorting them into separate X and Y populations that can subsequently be used for surgical intratubal or intrauterine insemination, deep-uterine insemination, regular artificial insemination in some cases, in vitro fertilization to produce sexed embryos for transfer, and intracytoplasmic sperm injection of ova. Skewed sex ratios of 85 to 95% of one sex or the other have been repeatably achieved in most species. The method has been used worldwide to produce several hundred morphologically normal animal offspring of the predicted sex. It has also been validated in the laboratory using DNA reanalysis of the sorted sperm populations and by fluorescence in situ hybridization and PCR of individual sperm. We developed a new orienting nozzle that we have fitted to both conventional and high-speed cell sorters that have been modified for sperm sorting. Recently we completed the adaptation of the new orienting nozzle to a Cytomation MoFlo high-speed cell sorter modified for sperm. This adaptation of the nozzle has increased the overall production rate of sorted X and Y sperm from about .35 million/h to 5 or 6 million sperm/h (each population). Calves have been born from cows artificially inseminated using conventional technique and sexed sperm. In addition, numerous litters of pigs have been born after transfer of embryos produced from X or Y sorted sperm. 相似文献
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Flow cytometrically sex sorted spermatozoa are reduced in their fertilizing capacity, particularly when stored either in cooling extender or after freezing in liquid nitrogen. So far, preservation methods for sorted spermatozoa have differed only marginally from procedures used for unsorted semen. In the present study, a TRIS extender was modified to balance major cell damage caused by the sorting process and by liquid storage of the sorted spermatozoa. The new extender, containing a combination of antioxidants (AO) and bovine serum albumin (BSA), significantly increased the lifespan and fertilizing capacity of sex sorted spermatozoa. No significant differences were observed between unsorted controls and sorted samples for motility and status of sperm membranes as tested by fluorescein-isothiocyanat-peanut agglutinin/propidium iodide (FITC-PNA/PI). Acrosome integrity of spermatozoa was significantly better when semen was stored at 15 degrees C for 24 and 48 h in an extender containing AO with or without BSA as compared with controls (p < 0.05). There were no significant differences, in pregnancy rates of heifers inseminated at a natural oestrus, between unsorted controls (16/24, 66.7%) and both sorted groups (AO + BSA: 18/31, 58.1% and AO-BSA: 12/22, 54.5%). Additionally, it was shown for the first time that artificial insemination (AI) with liquid sexed bull spermatozoa stored for 72 h after sorting can result in pregnancy rates similar to AI with non-sorted semen. 相似文献
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L An Z‐H Wu Y‐F Wu X‐L Zhang X Liu Y‐B Zhu W‐M Cheng H‐M Gao M Guo J‐H Tian 《Reproduction in domestic animals》2010,45(6):e344-e350
The present study was performed to test fertility in single‐ovulating and superovulated dairy heifers after insemination with low dose sex‐sorted sperm under field conditions. Some parameters, including the dosage, deposition site and timing, were assessed with the pregnancy rates after artificial insemination (AI). Moreover, the use of oestrus synchronization in combination with sorted sperm was evaluated. Besides that, we also improved the embryo production efficiency in superovulated dairy heifers by optimizing the timing of inseminations and repartitioning the sexed sperm dosage among multiple inseminations. The conception rate (52.8%) in heifers after low dose (2 × 106) insemination with sorted sperm deep into the uterine horn did not differ (p > 0.05) from that (59.6%) of conventional AI (1 × 107 non‐sorted sperm) and that of deep insemination with low dose non‐sorted sperm (57.7%). There was also no difference (p > 0.05) between conception rates after single (51.7%) and double (53.8%) deep insemination with sorted semen. Heifers inseminated with sorted sperm at synchronous oestrus had a lower pregnancy rate (48.1%) than heifers at spontaneous oestrus (53.6%), but this did not reach statistical difference (p > 0.05). The average number of transferable embryos collected in vivo from heifers inseminated with sorted sperm (4.81 ± 2.04) did not differ (p > 0.05) from that obtained from heifers after insemination with non‐sorted sperm (5.36 ± 2.74). Thus, we concluded that the pregnancy rate after deep intra‐uterine insemination with low dose sorted sperm was similar to that of non‐sorted sperm, which was either also deposited at a low dose deep intra‐uterine or into the uterine body. Sychronization of oestrus can be beneficial in combination with sorted sperm to optimize the organization and management of dairy herds. The results from superovulated heifers demonstrated that our insemination regime can be used to obtain a comparable embryo production efficiency with sorted sperm than with non‐sorted sperm. 相似文献
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R Bathgate N Mace K Heasman G Evans WMC Maxwell SP de Graaf 《Reproduction in domestic animals》2013,48(6):893-898
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. 相似文献
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Lindsey AC Morris LH Allen WR Schenk JL Squires EL Bruemmer JE 《Equine veterinary journal》2002,34(2):128-132
The objectives of this study were 1) to compare pregnancy rates resulting from 2 methods of insemination using low sperm numbers and 2) to compare pregnancy rates resulting from hysteroscopic insemination of 5 x 106 nonsorted and 5 x 106 spermatozoa sorted for X- and Y-chromosome-bearing populations (flow sorted). Semen was collected with an artificial vagina from 2 stallions of known acceptable fertility. Oestrus was synchronised (June to July) in 40 mares, age 3-10 years, by administering 10 ml altrenogest orally for 10 consecutive days, followed by 250 microg cloprostenol i.m. on Day 11. All mares were given 3000 iu hCG i.v. at the time of insemination to induce ovulation. Mares were assigned randomly to 1 of 3 treatment groups: mares in Treatment 1 (n = 10) were inseminated with 5 x 10(6) spermatozoa deposited deep into the uterine horn with the aid of ultrasonography. Mares in Treatment 2 (n = 10) were inseminated with 5 x 10(6) spermatozoa deposited onto the uterotubal junction papilla via hysteroscopic insemination. Mares in Treatment 3 (n = 20) were inseminated using the hysteroscopic technique with 5 x 10(6) flow sorted spermatozoa. Spermatozoa were stained with Hoechst 33342 and sorted into X- and Y-chromosome-bearing populations based on DNA content using an SX MoFlo sperm sorter. Pregnancy was determined ultrasonographically at 16 days postovulation. Hysteroscopic insemination resulted in more pregnancies (5/10 = 50%) than did the ultrasound-guided technique (0/10 = 0%; P<0.05) when nonsorted sperm were inseminated. Pregnancy rates were not significantly lower (P>0.05) when hysteroscopic insemination was used for sorted (5/20 = 25%) and nonsorted spermatozoa (5/10 = 50%). Therefore, hysteroscopic insemination of low numbers of flow sorted stallion spermatozoa resulted in reasonable pregnancy rates. 相似文献
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白绒山羊性控胚胎生产及移植应用研究初探 总被引：3，自引：1，他引：2
应用X性控冷冻精液对超排供体白绒山羊进行人工授精,生产性控胚胎并进行鲜胚移植。结果发现,①用性控冻精和鲜精输精的供体母羊,平均卵子回收数分别为13.3和12.5枚,其中性控冻精组受精卵比例为29.6%（55/186）,极显著低于鲜精组94.0%（281/299）的比例（P<0.01）;②性控胚胎和普通胚胎移植受体母羊产羔率分别为42.2% 和58.1% ,二者差异极显著（P<0.01）;③性控胚胎移植受体母羊所产羔羊雌性所占百分数为100%,极显著高于普通胚胎移植（P<0.01）。表明通过白绒山羊X、Y精子分离、人工授精生产性控胚胎,同时结合胚胎移植技术,可以达到按照生产实际中的意愿得到性别控制子代白绒山羊的目的。 相似文献
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Rath D Long CR Dobrinsky JR Welch GR Schreier LL Johnson LA 《Journal of animal science》1999,77(12):3346-3352
The objectives for the present experiments were to apply sperm sexing technology to an in vitro production system with porcine oocytes obtained from slaughterhouse material. On six experimental days, ovaries were obtained from an abattoir, and cumulus-oocyte-complexes were matured in vitro. Semen was collected from mature boars of proven fertility and was sorted for X-chromosome-bearing sperm, using the Beltsville Sperm Sexing Technology incorporating the use of high-speed sorting. A total of 5,378 oocytes were submitted for in vitro fertilization (IVF). Of these, 559 ova were stained for cytogenetic analysis 18 h after IVF. From the remaining 4,819 ova, 1,595 cleaved, and 1,300 of the cleaved embryos were transferred into 26 synchronized recipients (5 control gilts for unsorted sperm, 21 gilts for X-sorted sperm). In a test of two fertilization media (FERT-A vs FERT-B) higher cleavage rates (P<.05) were obtained when FERT-B was used as a fertilization medium for unsorted (43.4+/-5.1%) and sorted sperm (43.1+/-1.1%;), whereas in FERT-A unsorted sperm gave a cleavage rate of 17.9+/-4.4% and sorted sperm gave 30.4+/-1.4%. Additionally, cleavage rates were higher (P<.05) after fertilization with sorted sperm vs unsorted sperm, independent of fertilization medium. Cytogenetic analysis of ova revealed that more oocytes with unsorted than with sorted sperm remained in Metaphase 2 arrest (P<.05). This was also independent of the fertilization medium. Monospermic fertilization rates were the same for IVF with unsorted or sorted sperm, independent of the fertilization system, except FERT-A with unsorted sperm (P<.05). Polyspermic fertilization rates were highest in FERT-B (37.6+/-6.6). A total of 57 pigs were born from nine litters. Six litters from sexed sperm (X-sorted) produced 33 females (97%) and one male. Three litters from control transfers produced 23 pigs, 11 of which were female (48%). The sex ratio of the offspring was predicted based on the sort reanalysis of the sorted sperm for DNA content. 相似文献
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The objectives of this study were to determine whether calves produced by sexed sperm differed from controls and to what extent the sex ratio of calves was altered by the sexing procedure. Data were collected from 1,169 calves produced from sperm sexed by flow cytometry/cell sorting after staining with Hoechst 33342, and 793 calves produced from control sperm during breeding trials between 1997 and 2001. Least squares ANOVA were completed using factors of treatment (sexed vs. control sperm), 19 management groups from 13 field trials, and calf sex. Responses analyzed include gestation length, birth weight, calving ease, calf vigor, weaning weight, abortion rate, and death rates (neonatal and through weaning). No significant difference was observed for any response due to treatment or treatment interactions (P > 0.10). Therefore, calves produced from sexed sperm grew and developed normally both pre- and postnatally. A neurological disorder was observed in four control calves and one sexed calf from one farm. No gross anatomical abnormalities were reported for any calves in the study. Differences were observed for all responses among management groups (P < 0.03 for abortions and P < 0.01 for all other responses). Heifer and bull calves differed (P < 0.001) in gestation length (278.4 and 279.6 d), birth weight (32.8 and 35.2 kg), calving ease (1.15 and 1.30), and weaning weight (233 and 247 kg). Gestation length did not affect characteristics of calves. The sex ratio at birth of calves from unsexed control sperm was 49.2% male. Sexing accuracy of X-sorted sperm was 87.8% female calves, and Y-sorted sperm produced 92.1% male calves. Flow cytometry/cell sorting can be used to preselect sex of calves safely with approximately 90% accuracy. 相似文献