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精液冷冻技术在驴种质资源的长期保存和引进以及人工辅助繁殖技术应用等方面具有重要意义,精液冷冻保存有利于充分发挥优秀种公驴的繁殖力,降低驴养殖产业的生产成本。近年来,驴精液冷冻保护剂和冷冻方法不断优化,驴精液冷冻保存技术也取得了很大的进步。但由于驴冷冻复苏的精子活力和受胎率低,严重限制了驴冷冻精液的推广与应用。本文对驴精液冷冻保存技术的发展历程、精子冷冻损伤机制、影响驴精液冷冻保存的因素和驴冷冻精液的应用进行综述,为今后驴精液冷冻保存技术的研究提供参考。 相似文献
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兔精液冷冻保存对于家兔产业发展和种质资源保护具有重要意义。冷冻精液可以最大限度地保存优良种公兔的遗传基因,减少公兔饲养数量,同时也避免了活体引种时须进行的检疫、隔离饲养等程序,降低了兔场的饲养管理成本。此外,冻精保存作为活体保种的补充,在一定程度上缓解了种质资源“卡脖子”带来的潜在风险。但由于兔精子结构的差异,冻后精子品质下降、母兔受孕率降低,使得兔精液冷冻保存技术在实践生产中难以得到推广运用。本文概述了兔精子的结构与特点,对兔精液冷冻保存原理、影响因素、精液冷冻-解冻程序等研究进行综述,以期为兔精液冷冻保存技术的深入研究和应用提供参考。 相似文献
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本试验以不同浓度甘油对家兔精液冷冻保存的影响为研究目的,以柠-葡-卵基础液于三种浓度(6%、8%、10%)的甘油相配伍做为冷冻保存液对精液进行稀释,平衡,冷冻,解冻等处理,从而确定兔精液冷冻保存的最佳甘油浓度,得出如下结论:冷冻保护液中添加浓度为6%的甘油时,对精液进行冷冻保存、稀释、平衡,解冻后的精子活率与浓度为8%,10%的甘油冷冻效果比较差异显著. 相似文献
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A.A. Ponce V.A. Aires R. Carrascosa M. Fiol De Cuneo R.D. Ruiz J.L. Lacuara 《Research in veterinary science》1998,64(3):239
Chinchilla laniger is an endangered species and improved cryopreservation of spermatozoa would constitute a significant advance in the development of assisted reproductive techniques in this species. The functional activity of epididymal spermatozoa from adult males was studied immediately after extrusion and after 24 hours incubation, and the ability of five extenders to protect these gametes during cryopreservation was determined. A decrease in sperm motility, viability, acrosome intact cells and response to hypo-osmotic swelling test was detected 24 hours later. The extenders here assayed showed well-defined differences in their cryoprotective ability; however such differences could not be attributed to any one of their individual components. The presence of TES plus Tris, the proportion of the individual constituents and/or the differences in metabolic substrate content could explain the above-mentioned finding. The results indicate that cryo-buffer 11 (TES-Tris-egg yolk-fructose-glycerol) is the most powerful protector of sperm functional activity in this species. 相似文献
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Luna Gutiérrez-Cepeda álvaro Fernández Francisco Crespo Miguel ángel Ramírez Jaime Gosálvez Consuelo Serres 《Acta veterinaria Scandinavica》2012,54(1):72
Background
Variability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method. Different extenders and processing techniques (such us colloidal centrifugation) are used in order to optimize post-thaw sperm quality. Sperm chromatin integrity analysis is an effective tool for assessing such quality. The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols (prior to cryopreservation) in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times (i.e., their DNA fragmentation dynamics).Results
Post-thaw DNA fragmentation levels in semen samples subjected to either of the colloidal centrifugation protocols were significantly lower (p<0.05) immediately after thawing and after 4 h of incubation at 37°C compared to samples that underwent standard (control) centrifugation. The use of InraFreeze® extender was associated with significantly less DNA fragmentation than the use of Botu-Crio® extender at 6 h of incubation, and than the use of either Botu-Crio® or Gent® extender at 24 h of incubation (p<0.05).Conclusions
These results suggest that single layer colloidal centrifugation performed with extended or raw semen prior to cryopreservation reduces DNA fragmentation during the first four hours after thawing. Further studies are needed to determine the influence of freezing extenders on equine sperm DNA fragmentation dynamics. 相似文献13.
为了建立重庆板角山羊精液的细管冷冻保存方法,实验进行了不同冷冻稀释液(配方Ⅰ、Ⅱ、Ⅲ)、不同冷冻保存剂(甘油、EG)及不同离心速度(1000、1200、1400r/min)对重庆板角山羊细管精液冷冻保存效果的研究,结果表明:配方Ⅱ对重庆板角山羊精液的冻后活率显著优于配方Ⅰ和Ⅲ(P<0.05)。在配方Ⅱ中添加相同剂量(5%)的EG和甘油,精液冻后活率差异不显著(P>0.05)。以1200r/min的速度对山羊鲜精作离心处理后,冻后活率相对于对照组有所提高,但差异不显著(P>0.05)。 相似文献
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IB Lima‐Verde A Johannisson T Ntallaris E Al‐Essawe Z Al‐Kass T Nongbua F Dórea N Lundeheim K Kupisiewicz A Edman JM Morrell 《Reproduction in domestic animals》2018,53(1):127-136
Traditionally, extenders for bull semen included egg yolk or milk, but recently there has been a move to avoid material of animal origin. The aim of this study was to evaluate the effects of two commercial extenders (based on soya lecithin and liposomes) on bull sperm quality after cryopreservation. Post‐thaw sperm quality was evaluated by computer‐assisted sperm analysis and flow cytometric assessment of membrane integrity, chromatin integrity, mitochondrial membrane potential, production of reactive oxygen species and tyrosine phosphorylation. Furthermore, an artificial insemination (AI) trial was conducted, and 56‐day non‐return rates were evaluated. Semen frozen in the liposome‐based extender showed similar membrane integrity and higher mitochondrial membrane potential compared to those in the soya lecithin‐based extender. Chromatin integrity and production of live H2O2+ reactive oxygen species were similar in both extenders. Less superoxide was produced in the samples extended with liposome‐based extender, with or without menadione stimulation. Chromatin integrity and tyrosine phosphorylation were not affected by either type of extender. No differences in 56‐day non‐return rate between extenders containing soya lecithin and liposomes were observed in the AI trial (66% ± 0.8 and 65% ± 0.8, respectively). In conclusion, the sperm quality of bull semen frozen in the two extenders that do not contain material of animal origin was similar, although the semen frozen in the liposome‐based extender had higher mitochondrial membrane potential. Either extender could be used in situations where extenders containing material of animal origin are to be avoided. 相似文献
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Eva BLANCH Cristina TOMáS Marta HERNáNDEZ Jordi ROCA Emilio A. MARTíNEZ Juan M. VáZQUEZ Eva MOCé 《The Journal of reproduction and development》2014,60(2):143-149
Egg yolk (EY) and glycerol are common constituents of extenders used for sperm
cryopreservation. It has been demonstrated that using cholesterol-loaded cyclodextrins
(CLC) improves sperm cryosurvival in several species. However, standard freezing extenders
might not be the most appropriate for CLC-treated sperm. This study evaluated the EY and
glycerol requirements for freezing CLC-treated boar spermatozoa. Semen samples from 34
ejaculates coming from 4 boars were used. Each ejaculate was split into three aliquots:
one was used untreated (control), and the other two were treated with 1 mg of CLC or
methyl-β-cyclodextrin/120 × 106 sperm for 15 min at 22 C prior to
cryopreservation. Our results indicated that reducing the concentration of EY was
detrimental for sperm viability after thawing (31.57 ± 2 vs. 19.89% ± 2
for 20 and 10% EY, respectively; P <0.05), even in semen treated with CLC. On the other
hand, it was observed that the traditional concentration of glycerol (3%) was not the
appropriate for freezing CLC-treated sperm (61.10 ± 3 vs. 47.87% ± 3 viable sperm for
control and CLC-treated sperm, respectively; P <0.05). Thus, CLC-treated sperm showed a
higher tolerance to high glycerol concentrations (5%) in terms of sperm viability (59.19%
± 3) than non-treated sperm (45.58% ± 3; P<0.05). Therefore, it could be necessary to
modify the freezing extenders for CLC-treated sperm. Nevertheless, additional studies will
be needed to evaluate alternative cryoprotectants and to determine the effect of high
glycerol concentrations on sperm functionality. 相似文献
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E Jiménez CC Pérez‐Marín G Vizuete Y Millán EI Agüera 《Reproduction in domestic animals》2013,48(4):665-672
To evaluate and compare the efficacy of various extenders for the cryopreservation of epididymal cat spermatozoa, two experiments were planned. Bovine and equine commercial extenders in the experiment 1 and TRIS–egg yolk–based extenders in experiment 2 were separately studied since the number of sperm collected per cat is reduced. Epididymal sperm samples were packaged into 0.25‐ml straws and frozen. Vigour, motility, morphology, acrosome status, sperm viability and functional membrane integrity were assessed at collection, after cooling and after thawing, while DNA integrity was evaluated at 0‐ and 6‐h post‐thaw. Experiment 1 compared the effect of three non‐feline commercial extenders – based on TRIS–egg yolk (Triladyl), egg‐yolk‐free medium (AndroMed) and skimmed milk‐egg yolk (Gent) – on the quality of frozen‐thawed epididymal cat sperm. Values for sperm motility and functional membrane integrity in cooled sperm diluted in Triladyl were higher (p < 0.001) than those recorded for Andromed and Gent. Except sperm morphology, the other assessed characteristics showed significant higher values in frozen‐thawed sperm diluted in Triladyl than in Andromed and Gent extenders. Experiment 2 analysed the effects of three TRIS–egg yolk–based extenders, one non‐feline commercial (Triladyl) and the other two prepared using different monosaccharides (glucose and fructose), on freezing‐thawed sperm. Results showed that specifically prepared extenders for cryopreservation of feline spermatozoa performed better than the commercial extender Triladyl, although sperm quality during the freezing‐thawing process did not significantly differ associated with the type of monosaccharide (glucose vs fructose) added to the mentioned extenders. Although TRIS–egg yolk–based extenders prepared in experiment 2 improved sperm cryoprotection, Triladyl remains a good option for practitioners who, for ease of use and availability, prefer to work with commercial extenders. 相似文献
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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. 相似文献
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Antioxidant supplementation,effect on post‐thaw spermatozoan function in three sturgeon species 下载免费PDF全文
High levels of reactive oxygen species (ROS), which may be associated with reduced sperm quality, can be detected during cryopreservation of sperm of some species. Our objective was to investigate whether the addition of antioxidants to cryopreservation extenders influenced post‐thaw sperm characteristics and fertility in Acipenser dabryanus, Acipenser sinensis and Acipenser baerii. Prior to freezing, sperm samples were diluted with a base extender as control or in extender supplemented with catalase (CAT), glutathione (GSH), cysteine (NAC), ascorbic acid (VC) or their paired combinations. Protective concentrations of CAT, GSH and VC in the three species were 25 U/ml, 0.25‐0.5 mg/ml and 0.5 mg/ml, respectively. Cysteine showed no protective effect against ROS. The addition of CAT, GSH and VC positively affected either acrosome or membrane integrity of post‐thawed sperm in the three species, as well as spermatozoan motility in A. sinensis. The combination of antioxidants did not show a positive synergistic effect. This study suggested that the use of antioxidants in the cryopreservation of sturgeon sperm has potential to decrease intracellular ROS, and consequently preserve acrosome and membrane integrity, as well as spermatozoan motility. 相似文献
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JC Monteiro JSA Gonçalves JA Rodrigues CF Lúcio LCG Silva MEOA Assumpção CI Vannucchi 《Reproduction in domestic animals》2009,44(S2):359-362
Poor sperm viability post-thaw has resulted in constant research into methods of cryopreservation of canine semen. One factor that may be involved in poor viability is sperm oxidative stress caused by excessive formation of reactive oxygen species. The present study was performed in order to evaluate the effect of different concentrations of ascorbic acid (AA) and glutathione (Glu) added to an extender for the freeze-thawing of dog sperm. Semen from five mature dogs was collected and frozen in two studies. Prior to and after freezing, sperm motility, morphology and membrane status were examined. In addition, sperm motility was examined up to 120 min after thawing to evaluate thermo-resistance. In study I, semen was collected twice from each dog. On both occasions, semen was divided into three aliquots: control, Glu 1 m m and Glu 5 m m . In study II, semen was collected twice and divided into three aliquots; control, AA 50 μ m and AA 250 μ m . Initial sperm motility was significantly higher in sperm diluted with AA 50 μ m ; sperm longevity, however, measured by a thermal-resistance test (TRT), was higher for Glu treatments. Higher concentration of Glu produced significant improvement in TRT and membrane status, whereas higher concentration of AA had a negative impact in sperm longevity. Antioxidant supplementation to semen freezing extenders improved semen quality post-thaw. Moreover, Glu had the most beneficial effect when supplemented at 5 m m . 相似文献
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The aim of the present study was to determine the suitability of using two egg yolk-free commercial extenders, Andromed and Biociphos Plus as compared with the Tris-egg yolk based diluent Biladyl, for the cryopreservation of bull spermatozoa when the freezing protocol involved holding the extended semen at 4 degrees C for 18 h before the freezing. Six ejaculates from each of 10 Holstein bulls were collected by using artificial vagina. The ejaculates were evaluated for volume, sperm concentration and motility, divided in to three equal volumes, and diluted, respectively, with the three extenders as specified above. Extended semen was equilibrated for 18 h at 4 degrees C and frozen in 0.25-ml straws. After thawing, 100-mul aliquots of semen were labelled with SYBR-14, PI and PE-PNA (Phycoerythrin-conjugated Peanut agglutinin) and analysed by flow cytometry at 0, 3, 6 and 9 h after incubation at 37 degrees C. A General Lineal Model procedure for repeated measures was used to determine the effects of extender, bull, replicate and the interaction between them, on sperm viability and acrosomal integrity. Semen samples frozen with Biladyl showed higher (p < 0.001) sperm survival after 0 h (47.9%) and 9 h (30.3%) of incubation than those frozen with Andromed (38.5% and 17.3%, after 0 and 9 h respectively) or Biociphos Plus (34.9% and 21.6%, after 0 and 9 h respectively). The bull and replicate had significant effects (p < 0.001) on both sperm viability and acrosomal integrity, but the interactions between bull and extender and between replicate and extender were not significant. It was concluded that, when holding the semen overnight before freezing, the use of Biladyl results in higher sperm survival and longevity than the use of Andromed or Biociphos Plus. 相似文献