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1.
根据活精子对特定染料不着色而死精子易于着色,以及活精子在低渗环境中出现膨胀,死精子不能膨胀的原理。应用伊红低渗溶液对犬,猪,牛,鸡活精子的代谢能力进行评定,同时用常规精子评定法进行评定。二者进行差异显著性检验,差异均不显著。说明伊红低渗溶液对犬,猪,牛,鸡精子生活能力的评定是一种行之有效的方法。而且该方法简便,判断客观,具有实用性。  相似文献   

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<正> 前言 顶体反应是精子受精过程的重要步骤,也是估测精子受精能力的重要指标。本文就是从这个角度出发,采用TST三重染色法染色黑白花奶牛不同公牛个体解冻后的精子,观察精子的顶体反应率,同时测定了精子的活力,顶体完整率和畸形率,以及精清中钙的含量和乳酸脱氢酶(LDH)活性,从而来探讨TST三重染色法是否可作为评定精液品质的可靠方法。  相似文献   

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本文用TG染色检测德国牧羊犬冷冻精子顶体的形态。根据染色结果可将精子分为四类:a.有正常顶体活精子;b.无正常顶体活精子;c.有正常顶体死精子;d.无正常顶体死精子。冷冻精液效果存在着个体差异。顶体损伤率过高是导致犬冷冻精液受胎率低的主要原因。  相似文献   

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使用荧光染色和流式细胞术检测山羊精子   总被引:1,自引:0,他引:1  
分别使用PI(碘化丙碇),FITC-PSA(异硫氰酸荧光素络合豌豆凝集素)和RH123(罗丹明)对山羊冷冻精子进行染色,再使用流式细胞仪进行检测、分析。试验发现,对于山羊的冷冻精子,PI染色阴性的为质膜完整的精子;FITC-PSA染色阴性的可能为顶体完整的精子,染色FITC-PSA+和FITC-PSA++,可能分别为顶体反应中的精子和顶体破损精子;RH123+应为线粒体无活性精子,有绿色荧光可能是因为染料残留在细胞膜上导致,而RH123++可能应为线粒体有活性的活精子。不同个体羊的精子对于低温敏感度不一样。不同个体羊之间的冻精顶体完整率、质膜完整率、线粒体有活性的精子比率之间差异显著,据此应选择合适的山羊个体进行精液冷冻。  相似文献   

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对精液进行准确的分析 ,是任何人工授精方案中的关键步骤。显然 ,要试验精子的受精力 ,最好的方法就是对母猪进行输精 ,然后看结果如何。但是 ,如果结果不好 ,这时采取措施则已为时太晚。为了预先保证只将优质精液用于人工授精 ,我们就必须能在实验室内对精液样品进行分析。这样做 ,已经比较可行了 ,因为近年来已经开发出了多种多样的有关技术。应用这些技术 ,我们就有比较大的信心在实验室内对精液进行仔细检查 ,既可检查其物理结构 ,也可检查其受精能力。猪的精子是一种高度特化的细胞 ,它具有多种功能特性 ,在精液采集、储存、运输和输精…  相似文献   

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精子畸形率为评定猪精液品质的重要指标,精子畸形率≤20%才合格.精子畸形率过高将直接影响到猪人工授精的受胎率.笔者通过总结姬姆萨染色制片过程中的经验体会,提出了能够提高精子染色效果的方法,通过提高制片染色质量提高精子畸形率检查的准确度,能够减少由于制片染色不当造成的误判.  相似文献   

11.
Adverse Effects of Cadmium on Bull Spermatozoa   总被引:2,自引:0,他引:2  
Cadmium (Cd) is a widespread environmental pollutant. Because of its long biological half-life (10–30 years in humans), Cd accumulates in the biological systems such as gonads. The present study was designed to evaluate the effect of Cd in the concentration range 50–750 μmol/L, in vitro, on the membrane integrity, motility and acrosomal status of bull spermatozoa. The samples were processed for sperm analyses using semen-diluting fluid (phosphate-buffered saline, pH 7.2). A significant elevation in the malondialdehyde level/lipid peroxidation (LPO) rate and a decrease in the spermatocrit values, particularly at a concentration of 750 μmol/L Cd, indicated the deleterious effect of Cd on sperm membrane integrity. There was also a negative correlation between LPO rate and percentage of motile spermatozoa (r = 0.992).The gelatin test indicates that Cd may alter the integrity of acrosomal membranes and shows an abnormal acrosome reaction. In this regard, a strong negative correlation was found between LPO rate and % halos (bright clear zone around sperm heads after gelatin digestion) (r = 0.990). Taking the results together, Cd proved to be a potential toxicant in the category of environmental factors that induce membrane impairment, lower motility, and decrease the rate of acrosome reactions, leading to male infertility. Apparently, the presence of Cd in the environment and seminal plasma exerts a toxic effect on sperm cells. Arabi, M. and Mohammadpour, A.A., 2006. Adverse effects of cadmium on bull spermatozoa. Veterinary Research Communications, 30(8), 943–951  相似文献   

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The present investigation was performed to study the effect of freezing and thawing on boar spermatozoa. Thirty-one ejaculates from four boars were investigated after thawing in three different thawing diluents (seminal plasma, OLEP, isotonic glucose solution).From each ejaculate one sample of 1 × 109 spermatozoa was thawed in each of the thawing diluents. Each sample was examined in a thermoresistance test in which motility was stimulated with caffeine 30 min. and 3 hrs. after thawing. Furthermore, acrosome morphology and ASAT release from the spermatozoa were investigated for each sample. One ejaculate from the two most frequently used boars was examined by electron microscopy after thawing in each of the thawing diluents.Differences in the aspects studied appeared between isotonic glucose solution and the other two thawing diluents in the thermoresistance test, in the response to caffeine stimulation 3 hrs. after thawing and in the amount of ASAT released from the spermatozoa. The influence on the acrosome morphology varied between the thawing diluents, but the acrosomal alterations did not seem to be connected with the damage reflected by the thermoresistance test and by the measurement of extracellular ASAT activity.The ultrastructural investigation showed that all spermatozoa examined had some degree of ultrastructural alteration as compared with freshly ejaculated boar spermatozoa treated in the same way. This alteration could not be related to any of the thawing diluents.Of the various laboratory tests the thermoresistance test and the measurement of ASAT release are suggested to be sensitive indicators of sperm damage during freezing and thawing. These tests might be useful indicators of variations in sensitivity of spermatozoa to the freezing-thawing procedure.  相似文献   

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Membrane alterations in bull spermatozoa after freezing and thawing and after the process of in vitro capacitation and fertilization were studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Even if the majority of the spermatozoa exhibited intact membranes after freezing and thawing (90%), one could distinguish between 3 types of membrane defects depending of the different structures involved. The first type showed loss of plasmalemma over the entire acrosome. In the second category the anterior part of the outer acrosomal membrane exhibited a pronounced extension, but was covered by a partly intact plasmalemma. The last category consisted of spermatozoa with extensive vesiculation and disruption of plasmalemma and the outer acrosomal membrane. This type of defect could not easily be distinguished from a true acrosome reaction. The cumulus cells showed an active phagocytosis of both intact and acrosome reacted spermatozoa.  相似文献   

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本文报道了用咖啡因与肝素、钙离子载体(I-A)协同处理诱导马、驴、猪冷冻精子体外获能的研究结果.咖啡因与肝素、I-A均有协同作用.在肝素或I-A处理中添加咖啡因,不仅能提高穿透率,而且能促进卵内雄原核的形成和发育.精子先经洗涤,再用咖啡因与肝素或I-A协同处理,可得到良好的获能效果.  相似文献   

15.
The study was conducted to investigate the effect of relaxin on motility, acrosome reaction (AR), viability and utilization of glucose in fresh and frozen‐thawed bovine spermatozoa. Both semen samples were washed twice through centrifugation (5 min at 600 g), and preincubated for 1 h at 39°C for swim up. The swim‐up separated spermatozoa were resuspended in a sperm Tyrode's albumin lactate pyruvate (Sp‐TALP) medium containing 0 (control) and 40 ng/mL porcine relaxin and incubated for 0–6 h. Sperm motility was determined on the basis of movement quality examined by a phase contrast microscope. Sperm viability and AR were evaluated by using the triple staining technique. The incorporation and oxidation of 14C‐glucose was assessed by a liquid scintillation counter. Motility was improved (P < 0.05) in both fresh and frozen‐thawed spermatozoa by the addition of relaxin to the Sp‐TALP medium, whereas relaxin showed no significant effect on viability in either fresh or frozen‐thawed spermatozoa. The percentage of AR increased (P < 0.05) when fresh or frozen‐thawed spermatozoa were incubated with relaxin. In contrast, the incorporation and oxidation of 14C‐glucose increased (P < 0.05) in both kinds of spermatozoa incubated with relaxin. Thus the results demonstrated that the addition of relaxin to the Sp‐TALP medium increased the motility, AR and utilization of glucose in fresh and frozen‐thawed bovine spermatozoa.  相似文献   

16.
A rapid recombinant antigen-based latex agglutination test (LAT) has been developed to detect specific anti-leptospiral antibodies from human and dog sera. The recombinant LipL32 antigen developed and used for detecting the antibodies is specific in detection of the pathogenic serovars of Leptospira as the expression of the LipL32 antigen is restricted only to the pathogenic leptospires. The sensitized latex beads are stable and could be stored at 4°C for more than three months without showing loss of activity for both weakly and strongly positive samples. The test is found to be sensitive, specific and accurate as compared to the standard microscopic agglutination test (MAT). Moreover, the recombinant antigen-coated latex beads could detect the specific anti-leptospiral antibodies in the acute phase of the illness. The test is simple and inexpensive, and is rapid in the management of large numbers of patients.  相似文献   

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猪冷冻附睾精子与体外成熟卵母细胞的体外受精   总被引:2,自引:0,他引:2  
研究了不同解冻温度(39-41℃,58-60℃,69-71℃)和解冻方式(干解冻、湿解冻)对猪冷冻附睾精子解冻后的活力、存活指数以及受精能力的影响。结果表明,采用同一种解冻方式,经高温解冻的精液,虽然解冻后的活力较好,但活力却下降很快,精子的存活时间和存活指数均不如低温解冻的精液。采用同一种解冻温度,湿解冻所需的解冻时间相对短,解冻后精子活力相对低,但存活时间和存活指数相应优于对应的干解冻试验组。经低温解冻的精子,受精能力最强,精子的受精能力与解冻时间呈显著正相关(r=0.91613,P=0.0103).使用改良TCM199培养液,猪受精卵和颗粒细胞单层共培养能发育至早期桑椹胚。  相似文献   

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A rapid screening assay forCampylobacter fetus in bull semen was developed using the polymerase chain reaction (PCR) and restriction endonuclease analysis (REA) to complement isolation by culture. An oligonucleotide primer pair (C1/C2) from the hypervariable region of 16S rRNA ofC. fetus was used to amplify a 362 base pair fragment by PCR. The PCR/REA assay, which is completed in 10 hours, detected as few as threeC. fetus subsp.venerealis cells in experimentally infected raw bull semen and in semen diluted with milk or egg yolk Tris (EYT). All the strains tested, of both subspecies ofC. fetus, were amplified, as were some otherCampylobacter species. Restricting the amplified products byAluI differentiatedC. fetus from the other organisms. There was no visible product generated by PCR fromC. sputorum subsp.bubulus, a saprophytic organism found in the prepuce of bulls, or from seven other species of bacteria found in semen. A modification of the PCR assay, using another primer pair (C3/C2) and two temperature PCR cycling conditions, increased the probability of detectingC. fetus subsp.venerealis. PCR amplification followed by REA could be used to screen bovine semen rapidly forC. fetus. In most cases, sequencing of C1/C2 PCR generated products would be preferable for distinguishing between the two subspecies ofC. fetus.Abbreviations AI artificial insemination - bp base pair - Cff Campylobacter fetus subsp.fetus - Cfv Campylobacter fetus subsp.venerealis - EYT egg yolk Tris - MH Mueller-Hinton - PCR polymerase chain reaction - REA restriction endonuclease analysis - TE Tris-EDTA  相似文献   

20.
OBJECTIVE: To develop rapid (< 8 hour) tests using polymerase chain reaction (PCR) for the diagnosis of equine herpesvirus 3 (EHV3; equine coital exanthema virus), equine gammaherpesviruses 2 (EHV2) and EHV5, equine adenovirus 1 (EAdV1), EAdV2, equine arteritis virus (EAV), equine rhinitis A virus (ERAV; formerly equine rhinovirus 1) DESIGN: Either single round or second round (seminested) PCRs were developed and validated. METHODS: Oligonucleotide primers were designed that were specific for each virus, PCR conditions were defined and the specificity and sensitivity of the assays were determined. The application of the tests was validated using a number of independent virus isolates for most of the viruses studied. The PCRs were applied directly to clinical samples where samples were available. RESULTS: We developed a single round PCR for the diagnosis of EHV3, a seminested PCR for EHV2 and single round PCRs for EHV5, EAdV1, EAdV2 and RT-PCRs for EAV and ERAV. The PCR primer sets for each virus were designed and shown to be highly specific (did not amplify any recognised non-target template) and sensitive (detection of minimal amounts of virus) and, where multiple virus isolates were available all isolates were detected. CONCLUSION: The development and validation of a comprehensive panel of PCR diagnostic tests, predominantly for viruses causing equine respiratory disease, that can be completed within 8 hours from receipt of clinical samples, provides a major advance in the rapid diagnosis or exclusion diagnosis of these endemic equine virus diseases in Australia.  相似文献   

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