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1.
Urospermia is a major ejaculatory dysfunction affecting stallions. It has been thought that urine-contaminated semen should not be cryopreserved; however, on select cases, urine contamination of semen cannot be avoided. A recent study suggested that urospermic semen can be cryopreserved after cushion centrifugation and extension. Thus, this study aimed to assess the use of single-layer colloid centrifugation (SLC) to process frozen-thawed urine-contaminated stallion semen. Raw ejaculates (n = 55) from eight stallions were split into three groups: no urine, low (20%), or high (50%) urine contamination. Semen was extended 1:1, cushion-centrifuged, and resuspended at 200 million sperm/mL in BotuCrio. Resuspended semen was loaded in 0.5 mL straws and cryopreserved in liquid nitrogen. Samples were thawed (37°C for 30 seconds) and processed by SLC (400 g/30 minutes). Percentages of total motility (TM) and progressive motility (PM) were assessed with computer-assisted semen analyzer. Sperm viability (%VIAB) and yield were assessed with a NucleoCounter before and after gradient centrifugation. Data were analyzed with two-way ANOVA and Tukey’s test. The motility parameters TM before SLC (control: 35 ± 2; low: 33 ± 0.7; high: 22 ± 1.8) after SLC (control: 51 ± 3.6; low: 42 ± 2.2; high: 25 ± 2.8) and PM before SLC (control: 24 ± 1.8; low: 21 ± 1.14; high: 12 ± 1.5) and after SLC (control: 40.3 ± 3.2; low: 31 ± 3.9; high: 14 ± 2) significantly decreased with increasing urine contamination. Urine contamination marginally reduced (P < .05) sperm viability after cryopreservation before SLC (control: 45 ± 0.7; low: 27 ± 0.2; high: 27 ± 0.3) and after SLC (control: 54 ± 0.5; low: 49 ± 0.7; high: 38 ± 0.6). Recovery rates of sperm after centrifugation were not significantly different between groups. In conclusion, urine contamination affects sperm motility parameters in a dose-dependent manner. Post-thaw SLC selected sperm with higher motility and viability in control and low groups but only selected sperm with higher viability in the high group.  相似文献   

2.
Directional Freezing of Equine Semen in Large Volumes   总被引：5，自引：0，他引：5  
Despite its potential impact on the horse industry, sperm cryopreservation is not an established technology throughout the industry, for a number of reasons that include a reduction in pregnancy rate and increased cost per pregnancy. We have evaluated a novel directional freezing technique, based on a multi-thermal gradient (MTG), by comparing it with the conventional, controlled-rate cryopreservation method (CRCM). Ninety-seven ejaculates with > or =50% motility, collected from 31 stallions were each divided into two parts and subsequently frozen by either MTG or CRCM. Frozen samples were then stored in liquid nitrogen until thawing. The two treatments were evaluated by three methods: progressive linear motility (PLM), viability stain and hypoosmotic swelling (HOS) test. High correlation was found between the three evaluation methods for all post-thaw samples. Eighty-eight per cent of the ejaculates frozen by MTG had post-thaw PLM > or =35%, whereas only 59% of the ejaculates frozen by CRCM had such motility. Post-thaw evaluations of samples frozen by MTG and CRCM were: PLM - 50.2 +/- 1.5% and 37.4 +/- 1.5%, respectively; viability - 53.6 +/- 1.5% and 39.5 +/- 1.4%, respectively; membrane integrity, as evaluated by HOS - 36.2 +/- 1.3% and 26.5 +/- 1.1%, respectively. The differences according to all the evaluation methods were highly significant (p < 0.001), and the results indicate that freezing stallion semen by MTG is superior to CRCM.  相似文献   

3.
Electroejaculation in rhinoceroses has historically yielded inconsistent results, with the collection of high-quality, sperm-rich samples rare. The goal of this study was to develop a reliable method of electroejaculation in the rhinoceros by designing a rectal probe that appropriately fits the anatomy of this taxon and refining the procedure. A curved probe handle ending in an oblate, ellipsoid head was built using readily available supplies. A combination of rectal massage, penile massage, and electrical stimulation with a specially designed probe was employed in attempts to collect semen on 14 occasions from greater one-horned rhinoceroses (Rhinoceros unicornis; n = 4), black rhinoceroses (Diceros bicornis; n = 2) and a southern white rhinoceros (Ceratotherium simum; n = 1). During 13 of the 14 attempts, ejaculates were collected in multiple fractions. All but one of the ejaculates contained spermatozoa, and seven ejaculates contained good-quality fractions of semen (-60% sperm motility; > or =20 x 106 spermatozoa/ml) suitable for sperm banking and assisted reproduction procedures. Mean (+/-SEM) values for volume, pH, osmolality, and total sperm number for ejaculates containing good-quality fractions (98.2 +/-21.8 ml, 8.5+/-0.1, 290.4+/-6.7 mOsm, and 37.1+/-12.0 x 10(9), respectively) did not differ (P > 0.05) from those containing only poor-quality samples. Urine and/or erythrocyte contamination was not uncommon in fractions of both ejaculate types. Males producing good-quality samples ranged in age from 7 to 34 yr. None of the samples contained > or =75% morphologically normal spermatozoa. Electroejaculation with a uniquely designed probe consistently produced ejaculates in the rhinoceros. However, the production of high-quality samples continued to be challenging, occurring in only 50% of collection attempts. Regardless, the technology has progressed to a stage at which good-quality semen samples can be produced for sperm banking and assisted reproduction, and thereby can be integrated into intensive rhinoceros management strategies for the ultimate survival of this taxon.  相似文献   

4.
In swine, predicting the fertilizing ability of boar ejaculates before using seminal doses for artificial insemination purposes is very important for pork breeders. Routinely, semen quality is evaluated by means of sperm concentration, viability, motility and morphology. However, in some cases, these spermiogram parameters may not be precise enough to detect altered/non-functional spermatozoa within boar ejaculates that may yield lower reproductive performance. The present work reviews the conventional parameters most used for assessing porcine semen quality, and it also describes other markers recently found that may help for evaluating more accurately the boar sperm function and survival. These markers are related to alterations induced by defective spermatogenesis, epididymal maturation or sperm handling.  相似文献   

5.
Redig PT  Goyal SM 《Avian diseases》2012,56(2):411-413
Serum or plasma samples from raptors that prey or scavenge upon aquatic birds were tested by a commercially available blocking enzyme-linked immunosorbent assay for the evidence of antibodies to influenza A virus. Samples were taken from birds (n = 616) admitted to two rehabilitation centers in the United States. In addition, samples from 472 migrating peregrine falcons (Falco peregrinus) trapped on autumnal and vernal migrations for banding purposes were also tested. Only bald eagles were notably seropositive (22/406). One each of peregrine falcon, great horned owl (Bubo virginianus), and Cooper's hawk (Accipiter cooperi) from a total of 472, 81, and 100, respectively, were also positive. None of the turkey vultures (n = 21) or black vultures (n = 8) was positive. No clinical signs referable to avian influenza were seen in any bird at the time of capture. These data indicate that, among raptors, bald eagles do have exposure to influenza A viruses.  相似文献   

6.
Two experiments were conducted to evaluate the fate of sperm following uterine insemination. In Exp. I, five pairs of Holstein cows were inseminated with egg yolk-Tris extended semen (approximately 1.0 X 10(9) sperm; .5 ml) from five ejaculates from a single bull that had high levels (approximately 70%) of morphologically abnormal sperm. Cows were slaughtered 12 h after insemination. The genital tracts were removed and promptly clamped into defined regions. Sperm were recovered by flushing with 2.9% sodium citrate buffer. Proportions of abnormal sperm in the various regions were compared with those in the inseminate. Sperm numbers were also determined from each region. Regions of the tract varied in number of sperm (P less than .001), proportions of knobbed acrosomes (P less than .001), tapered heads (P less than .001), protoplasmic droplets (P less than .001), tail abnormalities (P less than .029) and total abnormalities (P less than .002). A total of 63.5 +/- 6.4 X 10(6) sperm was recovered. These sperm were distributed throughout the tract as follows: vagina, 91.8%; cervix, 5.4%; uterine horns, 2.7%, and uterotubal junctions-isthmi, .04%. No sperm were recovered from ampullae. Because retrograde movement of sperm from the uterus occurred in Exp. I, we conducted Exp. II to determine the extent of sperm loss from the genital tract following insemination. Three pairs of Holstein cows were inseminated with .42 X 10(9) sperm (.5 ml; egg yolk-Tris extender) from the same bull used in Exp. I (three ejaculates). All discharged mucus and urine was collected for 12 h after insemination for recovery of sperm. Aspirates (approximately 1 ml) of mucus from the vagina were evaluated during the 12-h post-insemination period for numbers of sperm and leucocytes. Sperm were also recovered from the tract following slaughter (approximately 12 h) to determine retention. Overall, 73 +/- 3.7% of inseminated sperm were recovered. Components were: inseminate lost from the genital tract in discharged mucus, 60 +/- 4.6%; lost in urine, .06 +/- .02%; aspirated from the vagina, 4.4 +/- 1%; adhered to equipment, 1.3 +/- .3%, and retained in the genital tract, 6.5 +/- 1.6%. Predicted numbers of sperm contained in discharged mucus 2 h post-insemination were greater (P less than .009) than at subsequent hours.(ABSTRACT TRUNCATED AT 400 WORDS)  相似文献   

7.
The phylogenetic order Squamata has many representatives that could benefit from the use of semen preservation as a tool for assisting conservation. To date, few studies have been made evaluating the potential for collecting and preserving semen from snakes. The objectives of this study were to characterize semen parameters of the corn snake (Elaphe guttata), including appearance, volume, concentration, sperm motility, and sperm morphology, and to determine the longevity of corn snake sperm motility stored at 4 degrees C. Single semen samples were collected from 22 adult corn snakes. The appearance of the corn snake semen was generally cloudy, and the color was white to tan. Corn snake spermatozoa initially exhibited a median motility of 92.5%. Corn snakes were found to produce small-volume ejaculates (median 0.01 ml). However, the overall concentration of the snake ejaculate was high (chi = 852 x 10(6) +/- 585 x 10(6) spermatozoa/ml). Morphologically, a mean of 75.7 +/- 9.3% of the sperm cells in an ejaculate were normal. Snake ejaculate with a white appearance had significantly higher sperm concentrations (chi = 1,859 x 10(6) +/- 1,008 x 106 sperm cells/ml; F = 15.74, P = 0.001) than tan ejaculates (chi = 601 x 10(6) +/- 439 x 106 sperm cells/ml). Sperm motility decreased significantly in samples that were stored at 4 degrees C for greater than 48 hr in a refrigerator or Equitainer I. This is the first study to characterize semen volume, appearance, and concentration; sperm motility; and sperm morphology in captive corn snakes. The information derived from this study can be used to develop a model for a collection, cooled storage, and shipping program for semen from endangered or threatened captive and wild snakes.  相似文献   

8.
Low egg fertility and hatchability is a common problem in captive populations of rockhopper penguins (Eudyptes chrysocome chrysocome). These conditions make sustaining a captive population challenging. A method for collecting and evaluating semen from rockhopper penguins was developed to assist in the evaluation of low egg fertility found in one captive population. Six adult male rockhopper penguins were conditioned to allow semen collection once a week from the start of breeding season until ejaculates no longer contained sperm. A total of 59 ejaculates was collected between 17 September and 31 December 2004. Forty-five of these samples were evaluated for volume, pH, sperm concentration, and sperm quality (motility, viability, and morphology). There was a large variation between individuals and between collections for each individual. The mean motility was 34.5% (+/- 22%). Mean volume of ejaculate was 0.23 ml (+/- .31 ml). Mean concentration was 16.9 x 10(6) sperm/ml (+/- 48.7 x 10(6) sperm/ml). Mean number of sperm per collection was 1.7 x 10(6) (+/- 4.2 x 10(6)). Mean percentage of living sperm was 82.9% (+/- 18.1%). Mean percentage of sperm with normal morphology was 82.1% (+/- 18.8%). Mean pH was 6.47 (+/- 0.49). During this season, only one of these males paired with a female. The pair produced one fertile egg, but the embryo died early in incubation. Male rockhopper penguins had low sperm concentration and low motility indicating that low male fertility may be contributing to the poor egg fertility rate. This work represents the first step in an ongoing study to improve captive breeding of rockhopper penguins.  相似文献   

9.
The objective of this case study was to investigate whether semen centrifugation and low-dose insemination techniques would improve fertility of an aged subfertile Quarter Horse stallion with low sperm concentration, motility, and morphology in ejaculates. Forty-five mares were bred by one of five treatments (n = 9 per group) using the entire ejaculate as follows: (1) Group Body: body insemination with ejaculate diluted 1:1 in TAMU extender; (2) Group Body-Cent: body insemination after centrifugation and re-suspension of sperm pellet to 1 mL in TAMU extender; (3) Group Horn-Cent: deep horn insemination after centrifugation and re-suspension of sperm pellet to 1 mL in TAMU extender; (4) Group Cent-Hys: hysteroscopic insemination onto the uterotubal papilla after centrifugation and re-suspension of sperm pellet to 200 μL in Kenney-Modified Tyrode’s extender; and (5) Group Dens-Hys: hysteroscopic insemination onto the uterotubal papilla after discontinuous density gradient centrifugation and re-suspension of the sperm pellet in 200-μL Kenney-Modified Tyrode’s extender. Pregnancy rates did not differ among treatment groups (P = .77). Semen centrifugation for low dose insemination did not appear to improve fertility of this subfertile stallion, despite use of entire ejaculates for each individual insemination dose.  相似文献   

10.
This study was conducted to investigate the predictive capacity of fertility and litter size of sperm head morphometric measurements when the ejaculates fulfilled the minimum requirements commonly used in artificial insemination (AI). Semen samples from 11 rabbits (77 ejaculates) were evaluated for sperm motility, abnormal spermatozoa and sperm head morphometry using computer automated sperm analysis system. Morphometric dimensions for length, width, area and perimeter were analysed. Only ejaculates with more than 70% of motility rate and <15% of abnormal sperm were used for AI. A total of 1031 individual AI were performed in commercial rabbitries. Our results showed significant differences among animals for all sperm head measurements. The mean values for fertility and litter size obtained were 68.4 ± 0.01% and 9.3 ± 0.1% respectively. To assess the predictive value of morphometric dimensions in fertility, a logistic regression analysis was applied. Moreover, multiple linear regression analyses were used to examine the relationship between litter size and sperm head morphometric parameters. Logistic regression analysis rendered a significant model between fertility and area and perimeter, explaining the 0.65% variation. Multiple linear regression analysis rendered a significant model between litter size and width, area and perimeter that explained the 1.3% variation. By conclusion, the sperm head morphometric parameters assay showed low potential to predict fertility and litter size when the ejaculates fulfilled the minimum requirements commonly used in AI (motility and abnormal spermatozoa) in rabbit.  相似文献   

11.
The aim of this study was to examine the incidence of sperm agglutinations and their relationships with sperm quality and fertility. Semen samples of 40 boars of an AI-station were investigated. Nineteen of the 40 investigated boars showed a constantly low (< 10% agglutinated sperm), 3 an intermediate (10-20%) and 6 boars a high level (> 20%) of agglutination in raw semen. The degree of agglutination in sperm samples of 12 boars varied distinctly during the investigation period. During summer more (P < 0.05) agglutinated sperm were observed (11.0 +/- 11.6%) than during winter (6.2 +/- 7.3%). There was no association between bacterial contamination and incidence of agglutinations (P > 0.05). After dilution in extender the percentage of agglutinated sperm decreased from 6.2 +/- 7.3% to 1.1 +/- 1.4% (P < 0.0001). Twenty-four hours after dilution the percentage of progressively motile sperm was 7.4% lower (P < 0.05) in ejaculates with an initially high degree of agglutination (> 20% agglutinated sperm) compared to samples with an initially low degree of agglutinated sperm (< 10%). Plasma membrane integrity, mitochondrial membrane potential, acrosome reaction and chromatin structure were independent (P > 0.05) from the level of agglutination. Fertility data did not differ (P > 0.05) between boars with low and high numbers of agglutinated sperm in raw semen. The results show that there are individual, ejaculatory and seasonal variations in the incidence and degree of agglutination. Agglutinations have a negative effect on motility of sperm and disappear to a large extent after dilution in sperm extender. They have no negative consequences on fertility.  相似文献   

12.
Identification of duck plague virus by polymerase chain reaction   总被引：33，自引：0，他引：33  
A polymerase chain reaction (PCR) assay was developed for detecting duck plague virus. A 765-bp EcoRI fragment cloned from the genome of the duck plague vaccine (DP-VAC) virus was sequenced for PCR primer development. The fragment sequence was found by GenBank alignment searches to be similar to the 3' ends of an undefined open reading frame and the gene for DNA polymerase protein in other herpesviruses. Three of four primers sets were found to be specific for the DP-VAC virus and 100% (7/7) of field isolates but did not amplify DNA from inclusion body disease of cranes virus. The specificity of one primer set was tested with genome templates from other avian herpesviruses, including those from a golden eagle, bald eagle, great horned owl, snowy owl, peregrine falcon, prairie falcon, pigeon, psittacine, and chicken (infectious laryngotracheitis), but amplicons were not produced. Hence, this PCR test is highly specific for duck plague virus DNA. Two primer sets were able to detect 1 fg of DNA from the duck plague vaccine strain, equivalent to five genome copies. In addition, the ratio of tissue culture infectious doses to genome copies of duck plague vaccine virus from infected duck embryo cells was determined to be 1:100, making the PCR assay 20 times more sensitive than tissue culture for detecting duck plague virus. The speed, sensitivity, and specificity of this PCR provide a greatly improved diagnostic and research tool for studying the epizootiology of duck plague.  相似文献   

13.
Computerized motility analysis (CASA) shows that four separate subpopulations of spermatozoa with different motility characteristics co-exist in rabbit ejaculates. There were significant (p < 0.01) differences in the distribution of these subpopulations among separate genetic lines, total sperm abnormalities and the percentage of altered acrosomes. Furthermore, logistic and linear multivariate regressions among several parameters of rabbit semen quality analysis were tested for use as predictive tools for the fertilizing ability of a specific artificial insemination semen sample. Logistic regression analysis rendered two mathematical, significant (p < 0.01) models: one between sperm viability and conception rate and the other between total sperm abnormalities and conception rate. Multiple linear regression analyses also yielded some significant relationships between both fertility (p < 0.001) and litter size (p < 0.05), with respect to some semen characteristics. Our results support the hypothesis that the predictive in vivo fertility use of the standard rabbit semen quality analysis coupled with a CASA determination could be reasonably achieved by applying linear and logistic regression analyses among several parameters of rabbit semen quality analysis.  相似文献   

14.
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   

15.
West Nile virus lineage 2 (WNV‐2) was detected in the brain of 17 goshawks (Accipiter gentilis) that succumbed to neuroinvasive disease in the Czech Republic during 2018: twelve birds were captive and five wild. Furthermore, two wild sparrowhawks (Accipiter nisus) and three other captive birds of prey (golden eagle Aquila chrysaetos, hybrid saker falcon Falco cherrug × F. rusticolus and Harris's hawk Parabuteo unicinctus) also died due to WNV encephalitis. The 2018 outbreak in Czech raptors clearly reflects a new epidemiological situation and indicates an increasing risk of both raptor and human infection with WNV‐2 in the country.  相似文献   

16.
Artificial insemination with frozen cauda epididymal sperm was performed in cats. Sperm were transmigrated from the epididymides in 10 male cats. The mean sperm motility and viability were 67% and 82.5%, respectively, and 11.6 x 10(7) sperm were recovered. The mean sperm motility after thawing was 24.0%. Eleven female cats received unilateral intrauterine insemination of 5 x 10(7) sperm, and the conception rate was 27.3% (3/11). This was the first case of conception obtained with frozen epididymal sperm in cats.  相似文献   

17.
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   

18.
The aim of our study was to compare the quality parameters of fresh feline ejaculates collected by three different techniques—urethral catheterization after medetomidine administration (CT), electroejaculation (EE) and epididymal slicing after orchiectomy (EP). A total of 34 adult male cats (Felis catus) were included in the study. In all male cats, the sperm collection was performed under general anaesthesia by three collection methods in the following order: urethral catheterization, electroejaculation and epididymal slicing. The sperm parameters evaluated were as follows: volume, motility, viability, sperm concentration, total sperm count and morphological examination. The highest quality semen parameters were achieved using EE. The comparison of results of the evaluated sperm quality parameters from EE and EP showed significant differences only in one case—the percentage of head abnormalities and lower percentage of head abnormalities were achieved using EE compared to EP: 8.5% (3.0%–21.0%) versus 10.0% (4.0%–22.0%). Semen collected by CT rendered the lowest quality samples when compared to sperm samples collected by EE and EP, especially with respect to the motility and total sperm count which were significantly lower (p < 0.001). Our study showed that sperm samples collected by EE and EP result in better quality of feline ejaculates compared to collection by CT from sperm samples collected from the same male cats. These results demonstrate the necessity of further research of urethral catheterization as a novel technique of semen collection in male cats.  相似文献   

19.
Basic characteristics of European bison (Bison bonasus) semen were described and the efficacies of two extenders-Triladyl, containing egg yolk, and a synthetic extender, containing soybean lipids-were tested for semen cryopreservation. Seven ejaculates were collected by electroejaculation from a 10-yr-old, European bison bull. Each ejaculate was diluted at 37 degrees C to a final concentration of 200 x 10(6) sperm/ml with Triladyl or the synthetic extender. Extended semen samples were frozen according to a standard bull semen freezing protocol. After 2 wk of storage, one straw from each extender and ejaculate was thawed, and postthaw quality was evaluated by individual sperm motility and movement rate, numbers of sperm morphologic abnormalities and intact acrosomes, functional integrity of the sperm membranes determined by hypoosmotic swelling test (HOST), viability (live-dead, eosin-nigrosin stain), and a heterologous in vitro sperm penetration assay (SPA). A total of 600 in vitro-matured bovine oocytes were inseminated with 1 X 10(6) spermatozoa of Holstein semen frozen-thawed in Triladyl (control) or of European bison semen frozen in Triladyl or the synthetic extender. Nuclear status of the oocytes was determined after 18 h of sperm-oocyte coincubation. Extender had no effect on any evaluated parameters of semen after dilution and cooling (4 hr at 5 degrees C) or in postthaw individual motility, quality of movement, and sperm morphology. However, significantly (P < 0.05) higher numbers of spermatozoa with intact acrosomes, intact membranes (HOST), and viable sperm (P < 0.01) were in semen frozen in Triladyl than in the synthetic extender. Mean values for heterologous SPA for bull (control) and for bison semen frozen in the synthetic extender were very much alike-63.3+/-10.6% and 63.1 +/- 15.9%, respectively; bison semen frozen in Triladyl was lower, 43.0+/-24.2% but not significantly different. Cumulative results from a variety of viability assays of diluted/cooled and frozen-thawed semen, including the heterologous SPA, suggest that European bison semen can be successfully frozen in both extenders tested in this study.  相似文献   

20.
In this study, two following experiments were performed to improve post-thaw motility and viability of frozen-thawed ram spermatozoa. We examined i) the effects of different concentrations of bovine serum albumin (0, 0.3, 1, 5, 10 and 15% BSA) in semen diluents lacking egg yolk and ii) the effects of four semen diluents, fructose (F: control) and trehalose (T) in semen diluents containing egg yolk, 15% BSA in semen diluents without egg yolk (BSA), and modified phosphate buffered saline (m-PBS). Frozen-thawed spermatozoa were examined for progressive sperm motility, viability, morphological abnormality, sperm tail swelling test, and sperm acrosome integrity. In Experiment 1, the rates of sperm motility immediately after thawing (0 h) were significantly (P<0.05) higher in the 10 and 15% BSA groups (55.0 +/- 2.9 and 58.3 +/- 6.7%, respectively) than in the positive control (F) group (41.7 +/- 4.4%). The rate of sperm viability in the negative control (0% BSA) group (80.2 +/- 3.3%) was significantly (P<0.05) lower than in the positive control (F) group (89.8 +/- 1.5%), but when compared with the F group, no significant differences were found among the 0.3, 1, 5, 10 and 15% BSA groups at 0 h. The rates of sperm morphological abnormality of the 10 and 15% BSA groups (6.5 +/- 1.3 and 6.3 +/- 1.1%, respectively) were significantly (P<0.05) lower at 0 h than that in the 1% BSA group (16.3 +/- 5.2%). In Experiment 2, T addition improved (P<0.05) the post-thaw motility compared with the F and BSA groups. Furthermore, at 3 and 6 h, the post-thaw motility of the T group (36.3 +/- 2.4 and 25.0 +/- 2.0%, respectively) was significantly (P<0.05) higher than in the BSA (26.3 +/- 2.4 and 18.8 +/- 1.3%, respectively) and F (28.8 +/- 3.8 and 18.8 +/- 2.4%, respectively) groups. The post-thaw sperm motility and viability in the m-PBS group were significantly (P<0.05) lower than those of the control (F), T, and BSA groups throughout all observation points. These results indicate that 10 and 15% BSA can be substituted for egg-yolk for ram semen diluent and that the addition of trehalose enhances motility and viability of ram spermatozoa after freezing and thawing.  相似文献   

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