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Contents A report is given on the deep-freezing of boar semen in pellet form at - 196° C. After thawing of the pellets a strong forward motility up to 45% was observed with the method described by NAGASE & NIWA (1964). The optimum conservation of the semen was obtained with a Tris extender.  相似文献   

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When ex vivo magnetic resonance (MR) imaging studies are undertaken, specimen conservation should be taken into account when interpreting MR imaging results. The purpose of this study was to assess MR changes during time in the anatomic structures of the equine digit on eight cadaver limbs stored at 4°C. The digits were imaged within 12 h after death and then after 1, 2, 7, and 14 days of refrigeration. After the last examination, four feet were warmed at room temperature for 24 h and reimaged. Sequences used were turbo spin echo (TSE) T1, TSE T2, short tau inversion recovery (STIR), and double-echo steady state (DESS). Images obtained were compared subjectively side by side for image quality and signal changes. Signal-to-noise ratio (SNR) was measured and compared between examinations. There were no subjective changes in image quality. A mild size reduction of the synovial recesses was detected subjectively. No signal change was seen subjectively except for bone marrow that appeared slightly hyperintense in STIR and slightly hypointense in TSE T2 sequence after refrigeration compared with day 0. Using quantitative analysis, significant SNR changes in bone marrow of refrigerated limbs compared with day 0 were detected in STIR and TSE T2 sequences. Warming at room temperature for 24 h produced a reverse effect on SNR compared with refrigeration with a significant increase in SNR in TSE T2 images. After 14 days of refrigeration a statistically significant decrease of SNR was found in bone marrow in TSE T2 and DESS sequences. The SNR in the deep digital flexor tendon was not characterized by significant change in SNR.  相似文献   

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This study was conducted to investigate the effect of glutathione-supplemented INRA82 extender on miniature Caspian stallion sperm quality during storage at 5°C. A total of 12 ejaculates from three stallions (four ejaculates from each stallion) were collected and diluted with INRA82 extender that included different concentrations of glutathione (0 [INRA-G0], 5 [INRA-G5], and 10 mM [INRA-G10]) and stored for 48 hours at 5°C. Sperm motility (computer-assisted sperm analysis), plasma membrane integrity (eosin–nigrosin staining) and functionality (hypo-osmotic swelling test), and malondialdehyde (MDA) level were determined during storage at 5°C. The results showed that the sperm total and progressive motility and plasma membrane integrity and functionality in all extenders were significantly decreased with increasing storage time. However, the MDA level in all extenders was significantly increased with increasing storage time. Also, the results showed that most of the evaluated sperm quality parameters in the present study, with the exception of MDA, were significantly greater in INRA-G5 than in INRA-G0 and INRA-G10 after 24 and 48 hours of storage at 5°C. We have concluded that supplementation of INRA82 with 5 mM glutathione can improve miniature Caspian stallion sperm quality during storage at 5°C by increasing total and progressive motility, plasma membrane integrity and functionality, and decreasing the MDA level compared with INRA-G0 and INRA-G10. More advanced in vitro evaluations and artificial insemination are required to reveal the exact effects of INRA-G5 on miniature Caspian stallion sperm quality and its fertilizing ability.  相似文献   

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This study was undertaken to investigate the effects of storage of stallion semen in a defined milk protein extender at 5 and 15°C under either anaerobic or aerobic conditions, with or without addition of the antibiotic gentamicin. Semen samples were collected from eight fertile stallions and stored for 96 h (day 0–4) and assessed daily for motility, velocity and membrane integrity (viability) using a CASA system. Samples for bacteriology assessment were taken on day 2 of storage. No significant (p > 0.05) differences in motility, velocity or viability were observed between treatments on days 0–2. On days 3 and 4, semen stored without gentamicin at 5°C had a significantly (p < 0.05) better semen quality compared with storage at 15°C without gentamicin, irrespective of oxygen exposure. On days 3 and 4, motility and velocity were greater in samples stored at 15°C with gentamicin, compared with the corresponding treatments without antibiotic (p < 0.05). This effect was also evident for viability on day 4. The decline in semen quality observed at 15°C most likely resulted from the effect of bacterial growth. Bacterial growth was the greatest in samples stored at 15°C without gentamicin, under both anaerobic and aerobic conditions (p < 0.05). Bacterial growth was inhibited by adding of gentamicin at 15°C, which accordingly reduced the decline in semen quality. Addition of antibiotic to samples stored at 5°C had no significant effect on any parameter analysed. In conclusion, storage at 15°C can be achieved by using an extender containing the antibiotic gentamicin. Storage at 5°C tended to maintain better semen quality irrespective of oxygen exposure, and did not necessitate an antibiotic treatment.  相似文献   

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To maintain the fertility of stallion spermatozoa during cooled storage, extender media are added to semen. In this study, three semen extenders were compared: EquiPro which contains defined caseinates and whey proteins instead of dried skim milk. The extender is provided in dry form and dissolved in distilled water prior to use. EquiPro TM has the same composition as EquiPro but is provided in a sterilized ready-to-use liquid form. AndroMed-E contains soybean lecithin as protein source. Semen was collected from seven stallions. Ejaculates were divided into three aliquots, diluted with the different extenders and stored at 5 degrees C for 4 days. Total motility, membrane integrity, average path velocity (VAP), curvilinear-velocity (VCL), straight-line velocity (VSL), distance average path (DAP), distance curved line (DCL) and distance straight line (DSL) were determined by computer-assisted analysis. Total motility decreased in all extenders during storage. The parameters VAP, VCL, VSL, DAP, DCL and DSL in semen diluted in EquiPro TM at most times and in semen diluted in AndroMed-E at some times were lower than in semen diluted in EquiPro (p < 0.05). Viability on days 0 and 4 was lowest in semen diluted in AndroMed-E (p < 0.05). Velocity decreased faster when semen had been diluted in the sterilized liquid extender EquiPro TM or in AndroMed-E compared with the dry formula of EquiPro. Therefore the liquid sterilized EquiPro despite no difference in its chemical composition differs from the dry, non-sterilized EquiPro extender. Heat sterilization apparently changes effects of the extender on spermatozoa.  相似文献   

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A system was designed to allow a small swine farm in a northern latitude to use its own boars for artificial insemination (AI) conveniently. Semen was collected twice weekly for 3 day use (days 0, 1 and 2), extended in an egg yolk extender and stored at 5°C. Farm personnel were trained to manage the entire AI programme. For simplicity all semen collected was used for insemination. In the first test 47 gilts and 15 sows were inseminated with semen from four boars. One boar was subfertile with a farrowing rate of 36%. The averages for the other boars ranged from 71 to 100%. Then semen was collected from seven boars and all was used to inseminate 70 gilts and 55 sows with 3 × 109 or more sperm. Overall 63% farrowed an average of 10.1 piglets per litter. Litter size for sows was 1.5 piglets larger than for gilts. There was no difference in farrowing rate when more than 3 × 109 sperm were inseminated. The feasibility of initiating a complete AI programme within a small herd using herd boars was established. However, selection of the boars, use of only high quality semen, and experience with detecting oestrus was required to increase the farrowing rate. The use of various agents to protect sperm against cold shock below 15°C is worthy of further investigation. A new type of electronic probe, which measures the conductivity of cervical mucus, could be helpful if a boar is not available for conventional detection of oestrus.  相似文献   

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Cooled stallion semen has a short viable life, which ranges with acceptable motility and viability from 24 up to 48 hours. The purpose of this study was to compare the effects of storage pH, the ability of three different zwitterionic buffers, and cholesterol-loaded cyclodextrins (CLC) to preserve the motility and integrity of stallion sperm cooled to 5°C for 48 hours. Fourteen ejaculates were collected and split to receive CLC or not (control group). After incubation, each sample was split into six subsamples and diluted in KMT extender containing 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES), or 2-(N-morpholino)ethanesulfonic acid (MES) buffers, and the final pH was adjusted to either 7.0 or 6.6, totalizing 12 experimental groups as a function of CLC, buffer, and pH variables (2 × 3 × 2 factorial). The motility parameters and integrity of plasma and acrosome membranes (live cell index) were determined using computer-automated semen analysis and epifluorescence microscopy at 3, 6, 24, and 48 hours of cooling period. According to results, pH was not a significant source of variation for motility and live sperm over different cooling periods. However, samples diluted in BES exhibited higher progressive motility within 3 hours and higher percentages of total motile cells after 48 hours of incubation at 5°C (P < .05). After 24 hours of storage, CLC-treated sperm samples presented higher motility than control group (P < .05), and after 48 hours of incubation, CLC-treated sperm exhibited higher percentages of live, motile, and progressively motile sperms (P < .05). We inferred that equine semen diluted in KMT containing BES as buffer and CLC treatment improve the equine sperm survival during storage at 5°C for 48 hours.  相似文献   

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The objective of this investigation was to establish the optimal extent of dilution for storage of boar spermatozoa at 15°C in Kiev diluent. The dilution titers used for the sperm-rich fraction of ejaculates from 8 boars ranged from 1:2 to 1:50. Seminal doses were stored for 10 days. Motility and acrosome morphology were evaluated after 1, 3, 5, 7, and 10 days of storage. The percentages of motile spermatozoa after 24 and 72 hours of storage were significantly higher for dilution rates between 1:7 and 1:11 than for dilution rates lower than 1:7 or higher than 1:11 (p < 0.05). The percentages of spermatozoa with normal acrosomes after 24, 72, and 120 hours of storage were significantly higher for dilution rates between 1:8 and 1:11 than for dilution rates lower than 1:8 or higher than 1:11 (p < 0.01 ).  相似文献   

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The aim of this study was to estimate the effects of Equex STM on sperm motion characteristics in chilled dog semen extended in Tris-based diluent. Thirty-two ejaculates were collected from 12 proven German shepherd stud dogs. The sperm-rich fractions were diluted in Tris-based extender with 1% (v/v) Equex STM (sample A) and in Tris-based extender with no addition of detergent (sample B). The extended semen was incubated for 240 h at 5°C and the motility parameters were evaluated by CASA system at 24-h intervals. Addition of Equex STM to Tris-based extender led to an initial activation of motion activity of spermatozoa, followed by a rapid decrease, shortening the lifespan of spermatozoa incubated at 5°C. Computer-assisted sperm analysis clearly showed that Equex STM-induced changes of sperm motion characteristics resemble the hyperactivation (HA) of spermatozoa associated with the capacitation process.  相似文献   

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In this work the role of energy substrates in the maintenance of boar-sperm survival during storage at 15-17 degrees C was tested. For this purpose, boar spermatozoa were stored at 15-17 degrees C in several defined media with separate combinations of a monosaccharide, glucose and a non-monosaccharide, either citrate or lactate, energy substrates. Our results indicate that the medium containing the highest concentration of glucose together with low lactate levels was the most suitable to maintain sperm quality for 168 h at 15-17 degrees C. This was confirmed after observation of the results of the percentages of viability and altered acrosomes, the osmotic resistance test, the hyperosmotic resistance test and the rhythm of L-lactate production. The survival ability of boar sperm was greater in this experimental medium than in the standard Beltsville Thawing Solution extender, which contains only glucose as an energy substrate, although at a concentration far higher than that of all the tested experimental media. Our results indicate that the exact composition, more than the pure quantity of energy substrates, is a very important modulatory factor which affects survival ability of boar sperm in refrigeration. Thus, the exact combination of several energy substrates would have to be taken into account when optimizing the design of commercial extenders to store boar spermatozoa at 15-17 degrees C.  相似文献   

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The purpose of this study was to evaluate the effect of cooling ovarian tissue on pig pre-antral follicles. Ovaries were maintained in saline solution (0.9%) at 4 or 20 degrees C for 6, 12 or 18 h. After storage, pre-antral follicles were morphologically evaluated. While primordial follicles were not affected by the storage, the percentage of morphologically normal growing follicles was significantly reduced in ovarian tissue stored at 20 degrees C for 12 or 18 h. To test the viability of stored follicles, growing follicles isolated from ovaries stored at 4 degrees C for 18 h and at 20 degrees C for 6 h were cultured for 3 days. Follicles stored in either condition presented the same growth pattern in vitro as fresh follicles. We conclude that storage of pig ovaries at 4 degrees C for up to 18 h or at 20 degrees C for up to 6 h does not affect the morphology of growing follicles or their ability to grow in vitro.  相似文献   

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This study was carried out to assess the in vitro quality of canine semen frozen in an ultrafreezer at -152 degrees C and to evaluate the male-to-male variation of frozen semen in five male dogs of the Canarian Mastiff breed. Four ejaculates of each dog were processed individually (5% glycerol and 0.5% Equex) to reach a final concentration of 100 x 10(6) spermatozoa/ml. Then, two freezing techniques were tested to assess the seminal quality (sperm motility, live spermatozoa and abnormal sperm cell percentages) at 1, 30, 60, 120 and 360 days after freezing: (i) semen was frozen and stored in liquid nitrogen; (ii) semen was frozen and stored in the ultrafreezer at -152 degrees C. After freezing-thawing, both freezing protocols showed no significant differences in sperm motility and the percentages of live and abnormal spermatozoa. On the other hand, the microscopic characteristics of spermatozoa in fresh semen were practically similar among males; however, after the semen processing and freezing, significant differences were observed (p < 0.05) among males, especially as regards sperm motility. This inter-individual variability was detected in both freezing protocols, showing that the male-to-male variation in the seminal quality post-freezing was independent of the freezing technique used. The in vitro results obtained in the Canarian Mastiff breed confirmed that the use of ultra-freezers at -152 degrees C is a potential alternative to liquid nitrogen for storing canine semen for long periods of time.  相似文献   

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