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1.
The aim of this work was to evaluate the effect of resveratrol (RSV) during liquid storage of stallion sperm for 24 hours at either 10°C or 4°C. The antioxidant RSV was added to reduce the oxidative damage that occurs during cold storage. Aliquots of 2 mL of diluted semen were stored either at 4°C or 10°C under anaerobic conditions, in the absence (control group) or presence of RSV at different concentrations (10, 20, 40, and 80 μM). Sperm quality parameters were assessed at 0 hours and after 24 hours of storage. Resveratrol treatment did not affect sperm quality parameters at 0 hours. At 24-hour storage, a significant (P < .01) decrease of sperm quality was observed independently from RSV supplementation and storage temperature. A significant decrease of viable spermatozoa with high mitochondrial membrane potential (SYBR+/PI−/JC-1+) was evident at 24-hour storage in 40- and 80-μM RSV groups compared with control group. Moreover, a decline of total motility in 80-μM RSV group compared with the control group and a decrease of progressive motility and average path velocity in 80-μM RSV group compared with control and 20-μM RSV groups were observed. In conclusion, our findings demonstrate that RSV supplementation does not enhance sperm quality of stallion semen after 24 hours of storage. Moreover, 40- and 80-μM RSV concentrations could damage sperm functional status, probably acting as pro-oxidant. Finally, although 24-hour storage significantly affected most of the sperm quality parameters, no significant differences were found in groups maintained at 4°C or 10°C, suggesting that stallion semen could be equally preserved at these different temperatures.  相似文献   

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Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is one of the cutting-edge methods currently applied in medical bacteriologic diagnostics. The aim of the study was to prove the possibility of applying MALDI-TOF MS to identify bacterial contamination in the ejaculate of stud stallions, which may cause infections to reproductive organs of mares following artificial insemination with cooled semen. A partial aim was to determine changes in the total count of microorganisms in long-term storage of ejaculate after its treatment with gentamicin and also without antimicrobial medication. Aerobic cultivation on Columbia agar was used to examine 26 semen samples from 13 horses; 31 different species of bacteria were isolated, which were identified by MALDI-TOF MS. The most frequently detected species came from Aerococcaceae, Staphylococcaceae, and Micrococcaceae families. The results of our work confirm that MALDI-TOF MS is a quick alternative method for identifying bacterial species that may contaminate stallion semen.  相似文献   

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Cooling of equine semen obtained from some stallions results in lower seminal quality and viability when the seminal plasma (SP) is present. The objective of this study was to evaluate the effect of the removal of SP using a Sperm Filter on the viability of cooled stallion semen. For this purpose, 31 stallions were used. Their ejaculates were divided into three groups: CN, semen was diluted with an extender; FLT, SP was removed by filtration; and CT, SP was removed by centrifugation and cooled to 15°C for 24 hours. Sperm kinetics and plasma membrane integrity were evaluated immediately after collection (T0) and after 24 hours of refrigeration (T1). No difference (P > .05) was noted at T1 for total sperm motility (TM), progressive sperm motility, or plasma membrane integrity when semen samples from all the stallions were analyzed. However, when samples from stallions termed “bad coolers” were analyzed (TM = <30% at T1), a difference was observed in TM and progressive sperm motility for CN compared with FLT and CT at T1. Sperm recovery was greater when SP was removed using the filter (FLT) to that when the SP was removed by centrifugation (CN) (89% vs. 81%). Thus, we concluded that filtering with a Sperm Filter is an efficient and practical method for removal of SP from stallion ejaculates, with lower sperm loss than centrifugation. We also found that the presence of SP reduces the quality and viability of cooled semen from stallions whose semen is sensitive to the process of refrigeration.  相似文献   

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本文研究了3种稀释液在0~5℃下保存火鸡精液的效果及稀释液pH值、稀释比例和卵黄浓度对精子存活力的影响。原精及稀释精液Ⅰ、Ⅱ、Ⅲ不保存时,受精率分别为71.7%,64.03%,63.29%和63.77%(P<0.05);在0~5℃下保存48h后,原精完全丧失受精率(0%),而稀释精液Ⅰ、Ⅱ、Ⅲ的受精率分别为52.40%,45.30%和20.33%(P<0.01)。火鸡精液在0~5℃保存过程中,pH值降低,原精比稀释精液降低程度大。在本研究条件下,pH6.5和1:3稀释最适合于火鸡精子的存活,稀释液中添加5%或10%卵黄可提高火鸡精子的存活力。  相似文献   

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Captive breeding has become an important tool in species conservations programmes, maintaining genetic diversity and restoring wild, endangered populations. In order to improve the reproductive efficiency of captive kept capercaillie, the purpose of the study was to determine the effect of selenium and vitamin E addition to semen extender on sperm characteristic during short‐term storage. Ejaculates collected individually from four capercaillie were divided into two parts, diluted threefold with basic EK extender and EK enriched with 1 mg/ml of organic selenium and 8 mg/ml of vitamin E (EK+Se+E) and stored 24 hr at temp. +4°C. Spermatozoa morphology, motility and motility parameter were evaluated in net, diluted and stored semen samples. Significant (p < .05) differences between individual males were stated in relation to the majority of traits evaluated in the freshly collected semen. Comparing to the fresh semen, a significant (p < .05) decrease in percentage of live sperm in total (by 3.8% points on average) has been observed in samples diluted by EK extender, while in semen diluted with EK+Se+E extender this decrease was lower (1.5%pts on average) and not significant. Also per cent of motile sperm in EK+Se+E extender was higher (p < .05) then in EK (71.6% vs. 58.9%), but taking into account the values of individual males, both extender and male effect on liquid semen storage become apparent. Obtained data allow concluding that selenium and vitamin E addition to EK extender had positive effect on morphology and motility of capercaillie semen stored 24 hr at 4°C and can be recommended for similar studies carried out on other Galliformes species.  相似文献   

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The addition of antioxidants to cryopreservation media reportedly improves sperm post-thaw quality and reproductive performance after artificial insemination. Therefore, the objectives of this study were to evaluate if the addition of L-carnitine and pyruvate to freezing media, or their addition to samples after thawing, improves the post-thaw quality of equine spermatozoa. Thus, in Experiment 1, stallion semen samples were cryopreserved in: (1) EDTA–glucose–based extender with 20% egg yolk and 5% dimethylformamide (EDTA control); (2) skim milk–based extender with 20% egg yolk and 5% dimethylformamide (milk control); (3) Extender 1 supplemented with 50 mM L-carnitine and 10 mM pyruvate (EDTA-carnitine-pyruvate); and (4) Extender 2 supplemented with 50 mM L-carnitine and 10 mM pyruvate (milk-carnitine-pyruvate). In Experiment 2, 50 mM L-carnitine and 10 mM pyruvate were added post-thaw to samples cryopreserved with extenders 1 and 2 (EDTA control and milk control). Sperm kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability were evaluated after thawing. No significant differences (p > 0.05) were observed for most of the kinematic parameters, DNA fragmentation, membrane lipid peroxidation, acrosome status and viability of spermatozoa, between the samples frozen in the presence or absence of L-carnitine and pyruvate, nor between the samples after the post-thaw addition of these components. A higher (p < 0.05) mean velocity and higher (p < 0.05) amplitude of lateral head displacement were observed in the samples frozen in the milk-based extender with the addition of L-carnitine and pyruvate after thawing. The addition of 50 mM L-carnitine and 10 mM pyruvate, either to the freezing extenders or after thawing, was not deleterious for sperm; however, it did not improve equine sperm motility, viability, acrosome and DNA integrity, nor decrease membrane lipid peroxidation after thawing.  相似文献   

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The objective of the study was to investigate if reducing the seminal plasma of stallion extended semen by centrifugation once will suffice to maintain acceptable semen quality for insemination after 4 days of cool storage. Collected semen was extended to 25 × 106 sperm/mL and subjected to one of the following treatments: noncentrifuged (control), centrifuged for 10 minutes at 900 × g and 1800 × g. The supernatant was partially removed, and the sperm pellet, reconstituted and re-extended. It was then placed in a passive cooling device overnight and then transferred to a refrigerator for the remainder of the cooling period. At day 0, 2, and 4, total motility (TM), progressive motility (PM), and plasma (PLM) and acrosomal membrane integrity were assessed. Centrifuged groups had higher TM and PM at day 4 than the control group (P < .05). Likewise, centrifuged groups had higher intact PLM in day 4 (P < .05). A single centrifugation cycle to reduce seminal plasma concentration will suffice to preserve sperm integrity acceptable for an artificial insemination dose up to 4 days of cool storage.  相似文献   

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The reproductive management of mares for frozen semen artificial insemination (AI) can be costly and labor intensive. Predicting the exact time of ovulation can be challenging even when ovulation-inducing drugs are used. The main objective of this retrospective study was to determine whether there was an effect of interval between examinations to detect ovulation on likelihood of pregnancy and early embryonic loss in mares after postovulatory breeding with a single straw of frozen/thawed semen. The second objective was to determine the efficacy of two different drugs (human chorionic gonadotropin vs. buserelin) for timely induction of ovulation. The length of the interval from penultimate check to ovulation had no significant effect on pregnancy or embryo loss rates (4 hours: 34.1% and 13.3% vs. 8 hours: 26.1% and 0% and 16 hours: 34.5% and 10%, respectively) nor did the ovulation-inducing drug used, number of the cycle, or the stallion. In conclusion, there appears to be no advantage of checking mares for ovulation during the late evening and night hours when using a postovulatory AI protocol and ovulation-inducing drugs.  相似文献   

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保存温度和不同种类稀释液对猪精液品质的影响   总被引:2,自引:0,他引:2  
猪精液在室温保存时,含庆大霉素的稀释液保存效果优于含青、链霉素的稀释液,两种稀释液在保存72h时,精子活率、顶体完整率、GOT和LDH活性以及存活指数差异极显著(P<0.01).低温保存时,含蜂蜜和卵黄的稀释液对防精子冷刺激、维持精子活率的效果比含葡萄糖、奶粉的稀释液理想.在冷冻保存条件下,以含2%甘油的稀释液保存效果最佳.液态保存(室温、低温)的猪精液,LDH活性随精子活率下降而下降;pH值则都呈先上升后下降的变化趋势.室温保存主要是引起精子顶体脊膨大和保存后期的顶体前端膨大;低温保存主要引起顶体前部膨大和保存后期的少数顶体脱落;冷冻保存主要是造成顶体的膨大程度加大和顶体脱落.3种保存条件都造成精子线粒体透性增强.液态保存和冷冻保存精液的精清GOT活性与顶体完整率无显著相关性(P>0.05).  相似文献   

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鸵鸟精液品质及精液低温保存条件的研究   总被引:1,自引:0,他引:1  
测定鸵鸟精液生理指标 ,结果表明 ,鸵鸟的射精量为 0 1 7~ 2 0 3mL ,精液 pH6 7~7 3,精子密度为 1 5 7~ 48 0亿 /mL ,原精液精子畸形率 3 0 9%~ 1 3 3% ,精子活率范围0 78~ 0 92。对鸵鸟精液用不同稀释液稀释后低温保存效果比较研究 ,初步获得在 0~ 5℃时用改进MEM液作稀释液以 1∶2比例稀释后 ,精子在设计的条件下存活时间平均约 72h ,而相应原精液或 0 85 %NaCl液作 1∶2稀释的精液在同样条件下精子仅能存活约 2 4h  相似文献   

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为了开发猪精液常温保存新型稀释粉,通过检测精子活率、质膜完整性、顶体完整性以及超氧化物歧化酶(SOD)活性、过氧化氢(H2O2)和丙二醛(MDA)产生量等指标,分析常温保存稀释粉对猪精液的保存效果的影响。结果表明,在常温保存条件下,自配稀释粉保存猪精液5d时间内,精子活率平均为0.63,精子顶体完整率和质膜完整率分别达到78.4%和48.6%;SOD活力、H2O2产生量和MDA产生量分别是177.39IU/mL、86.28mmol/mL和9.36nmol/mL,可以在生产实践当中应用。  相似文献   

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Peptostreptococcus anaerobius is a genus of anaerobic Gram + cocci with increasing resistance to the most commonly used antimicrobials. The purpose of this report is to present the successful management of bacteremia due to P. anaerobius in an adult horse. A 19-year-old, Westphalian stallion presented with chronic cough, intermittent pyrexia, depression, and partial anorexia. Laboratory examination revealed neutrophilic leukocytosis, thrombocytopenia, increase of alkaline phosphatase, and hepatic enzymes. The stallion was unresponsive to the initial supportive treatment presenting gradual increase in neutrophilic leukocytosis and azotemia. Ultrasonographic examination and serologic testing were performed to limit differential diagnosis. Blood cultures revealed P. anaerobius, and the antimicrobial regimen was modified accordingly. The horse responded positively to the aforementioned protocol, showing marked improvement of clinical signs and normal WBC values, 7 days after initiation of oxytetracycline therapy. To the best of our knowledge, this is the first report of bacteremia due to P. anaerobius in an adult horse and emphasizes the need of injudicious administration of antimicrobial agents, facing the challenge of multidrug-resistant anaerobic Gram + cocci.  相似文献   

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精子活率是精子的重要特性,传统精子活率的评价方法并不代表精子在家禽机体内生殖道的真实运动情况,具有主观性的特点,准确性差。精子迁移率Mobility的测定方法模拟精子在母鸡体内液体环境和温度下的运动情况,能够反映精子在体内真正的活力。从37只纯种白洛克公鸡中挑选出具有不同精子迁移率Mobility的高、低2组,每组5只公鸡,其精子迁移率分别为1.103,0.219,相应的受精率分别为83.7%、76.8%(P<0.05)。结果表明:精子迁移率Mobility反映了不同家禽个体精液质量的差异,是预测人工授精效果的一个重要指标。  相似文献   

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Although several selection techniques are available for processing spermatozoa, only colloid centrifugation has been used to any extent in this field, starting with density gradient centrifugation and progressing more recently to single-layer centrifugation (SLC). SLC through a species-specific colloid has been shown to be effective in selecting spermatozoa with good motility and normal morphology from stallion semen. The method is easier to use and less time-consuming than density gradient centrifugation, and has been scaled-up to allow whole ejaculates to be processed in a practical manner. The potential applications of SLC in equine breeding are as follows: to improve sperm quality in artificial insemination doses for “problem” ejaculates, to increase the shelf life of normal sperm doses, to remove pathogens (viruses, bacteria), to improve cryosurvival by removing dead and dying spermatozoa before freezing or after thawing, to select spermatozoa for intracytoplasmic sperm injection, and to aid conservation breeding.  相似文献   

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