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PubMed2003

Some aspects relating to the evaluation of the effects of chemicals on male fertility.

Reviews and studies on individual compounds were analyzed as to the suitability of different study designs and endpoints for detecting adverse effects of chemicals on male reproduction in animal species. Of the endpoints investigated, the most sensitive proved to be histopathology of the testes. Using refined histopathology, effects could be detected with a high degree of sensitivity as early as 4 weeks after treatment. Other sensitive endpoints were the weights of reproductive organs, including accessory glands, i.e., testis, epididymis, prostate, and of the seminal vesicle, as well as sperm parameters such as sperm count, sperm morphology, and sperm motility. Sperm motility was found to be in some cases more sensitive than histopathology. The above parameters showed a higher sensitivity than fertility parameters. In fact, in most cases, not only one but several endpoints were affected. Continuous breeding studies and 90-day studies with additional measurements of sperm parameters were similarly effective in detecting compounds which affect male fertility. Interspecies extrapolation factors (IEFs) have been derived for the most sensitive endpoints in laboratory animals. If the calculation is based on caloric demand and a sensitive endpoint of reproductive toxicity, many IEFs tend to be about 1, indicating that humans are generally not more susceptible to reproductive toxicants than laboratory animals. With respect to hazard identification, it is possible to detect adverse effects on male reproduction in a standard subacute study with concentrations that produce significant general toxicity. If effects are found, for the risk assessment the NOAEL has to be determined by testing specific sensitive parameters as specified above.

Animals↗
PubMed1987

Effect of oocyte quality and sperm characteristics on the number of spermatozoa bound to the zona pellucida of human oocytes inseminated in vitro.

Sperm characteristics and oocyte quality may play a role in in vitro fertilization. The objective of this paper is to analyze the effect of the quality of oocytes, the husband's semen characteristics, and category of the couple's infertility on the number of spermatozoa bound to the zona pellucida. One hundred eighty-one oocytes which failed to fertilize or failed to cleave were fixed in 2.5% glutaraldehyde 40 to 60 hr after insemination in vitro and examined under interference microscopy and the number of sperm bound to the zone pellucida was determined. The means +/- SD of sperm bound to mature, immature, and atretic oocytes were 51.0 +/- 50.7, 7.3 +/- 12.1 10.4 +/- 17.8, respectively. Fertilized mature oocytes (81.0 +/- 53.3) had a significantly higher number of sperm bound to zonae compared to unfertilized oocytes (41.8 +/- 47.3). It is concluded that the number of sperm bound to zonae is functionally important. The sperm motility and the number of motile sperm used to inseminate oocytes were significantly correlated with the number of sperm bound to zonae, whereas sperm morphology and sperm concentration did not correlate. This study supports the notion that sperm motility is the single most important factor influencing fertilization of human oocytes in vitro.

Female↗
PubMed2006

Determination of the diagnostic value of the resazurin reduction assay for evaluating boar semen by receiver operating characteristic analysis.

AIM: To assess that metabolic status of spermatozoa could provide a useful tool for evaluation of semen quality. METHODS: The accuracy of the spectrophotometric application of the resazurin reduction assay was assessed using receiver operating characteristic (ROC) analysis. RESULTS: Areas under ROC curves (AUC) for motile sperm concentration and sperm index (SI) (sperm concentration multiplied by the square root of percentage sperm motility multiplied by the percentage normal sperm morphology) were 0.922. The best discrimination between poor and good semen samples according to the SI was achieved at a cut-off point of A610 = 0.209, where high sensitivity (94.1%) and specificity (91.7%) were calculated. The assay was less accurate when motile sperm concentration was used as the criterion value, yielding sensitivity of 88.2% and specificity of 87.5%, respectively. Likelihood ratios (LR) indicate that absorbances lower than 0.209 were at least 11.3 times as likely to be found in good semen samples than those in poor according to the SI, whereas in the case of motile sperm concentration, the LR was calculated to be 7.06. CONCLUSION: These results show that the resazurin reduction assay combined with spectrophotometry is an accurate method of assessing the quality of boar semen.

Animal Husbandry↗
PubMed1986

In vivo and in vitro evaluations of spermatotoxicity induced by 2-ethoxyethanol treatment.

2-Ethoxyethanol (EE), a member of the glycol ethers, has been shown to produce testicular atrophy in laboratory animals. The present study further identified the spectrum of effects on the male reproductive system in vivo and initiated in vitro studies on possible mechanisms of action. Adult, male rats were treated (po) with 0 or 936 mg EE/kg, 5 days/week for 6 weeks. Semen samples were collected on a weekly basis during the exposure period from ovariectomized, hormonally primed females immediately following mating and analyzed for sperm count, sperm morphology, and sperm motility. Sperm count and percent normal morphology were decreased at Weeks 5 and 6, and sperm motility was decreased at Week 6. These data, along with other studies, indicated that the pachytene spermatocyte was the most sensitive target for EE. In vitro studies monitored O2 consumption and ATP concentrations in isolated pachytene spermatocytes which were treated with 10 mM EE or 1 or 10 mM ethoxyacetic acid (EAA), the reported active metabolite of EE. An increase in respiratory ratio for the lactate rate/endogenous rate and a decrease in the 2,4-dinitrophenol rate/lactate rate were observed only for cells treated with 10 mM EAA. Additionally, a decrease in ATP concentration was seen only with 10 mM EAA. These results indicated that EAA interfered with energy metabolism in the pachytene spermatocyte. This effect may, in part, explain the testicular toxicity produced by this compound.

Acetates↗
PubMedDate not supplied

Seminal fluid findings in men with nonbacterial prostatitis and prostatodynia.

There is considerable confusion about the effects of prostatitis syndromes on male reproductive physiology. Therefore, we correlated findings on seminal fluid and expressed prostatic secretions from 100 men attending a special prostatitis clinic. These men had symptoms of prostatitis but no evidence of urethritis, acute or chronic bacterial prostatitis, or significant urological abnormalities. All subjects were evaluated following a standardized protocol, including lower urinary tract localization studies, expressed prostatic secretion analyses, and seminal fluid analyses with Bryan-Leishman staining. Seminal fluid findings were compared in men with inflammation (> or = 10(6) leukocytes/ml) in their expressed prostatic secretions, i.e., nonbacterial prostatitis, and men without inflammation in prostatic secretions, i.e., prostatodynia. Of 23 men with inflammation (> or = 10(6) leukocytes/ml) in their seminal fluid, 6 (26%) had nonbacterial prostatitis (mean leukocyte concentration 8.6 +/- 9.4 x 10(6)/ml of semen) and 17 (74%) had prostatodynia (mean leukocyte concentration 6.2 +/- 7.0 x 10(6)/ml, not significant). Of 77 men who did not have seminal inflammation, 15 (19%) had nonbacterial prostatitis (mean leukocyte concentration 0.1 +/- 0.2 x 10(6)/ml) and 62 (81%) had prostatodynia (mean leukocyte concentration 0.1 +/- 0.2 x 10(6)/ml, not significant). Men with nonbacterial prostatitis had lower values for several parameters associated with sperm motility, especially the proportion of motile sperm (45% compared with 60% for men with prostatodynia, P = 0.08) and sperm subjective speed score (median 3 compared to 4 for men with prostatodynia, P = 0.03). In summary, a minority of men had seminal inflammation, even among men with nonbacterial prostatitis. There was poor correlation between inflammation in the prostatic secretions and in the semen. Nonbacterial prostatitis, but not seminal inflammation, was associated with reduced sperm motility. Our findings highlight technical issues and the importance of investigating different sites and samples, including the urethra, expressed prostatic secretions, and seminal fluid.

Adolescent↗
PubMedDate not supplied

Relationship between intracellular calcium, energy metabolism, and motility of ram sperm.

Collective sperm motility is characterized in terms of periodic aggregation or cooperation among cells, which are seen under a microscope as a continuous wave motion. In this study, sperm motility that was made dependent on mitochondrial activity (glycolysis inhibited) was significantly inhibited by mM exogenous calcium while glycolytic-dependent (mitochondrial respiration inhibited) motility was not inhibited under these conditions. At intracellular Ca2+ above 400 nM, sperm motility was inhibited independently of the source of ATP. At [Ca2+]i of approximately 110 nM, mitochondrial- or glycomitochondrial-dependent motility showed 75 or 0% inhibition, respectively, indicating higher sensitivity of mitochondrial-dependent motility to [Ca2+]i in comparison to glycolytic-dependent motility. Under the latter conditions, intracellular level of ATP was reduced by 16% only, indicating that the 75% inhibition of mitochondrial-dependent motility was not due to reduction in ATP. The inhibition of mitochondrial-dependent motility by mM extracellular Ca2+ can be prevented by incubating the cells at pH 6.5 instead of 7.6. At pH 7.6, calcium probably interacts with negative sites on the cell surface and interferes in cell-to-cell interactions, which are important to achieve collective motility. At more acidic pH (6.5) these negative sites are probably protonated; thus Ca2+ cannot interact with them and no inhibition of motility could be seen. Glycolytic-dependent motility was not inhibited by extracellular Ca2+, since the pH of the medium becomes acidic under these conditions. Like Ca2+, mersalyl, which interacts with Ca2+ binding sites on the cell surface, showed significant inhibition of mitochondrial-dependent motility without any effect on glycolytic-dependent motility. Collective motility was directly correlated with fertility. These data are significant for establishing better conditions for spermatozoa treatment when artificial insemination or in vitro fertilization is concerned.

Adenosine Triphosphate↗
PubMedDate not supplied

Comparison of sperm quantity and quality in antegrade V retrograde ejaculates obtained by vibratory penile stimulation in males with spinal cord injury.

Reduced fertility in men with spinal cord injury results from the inability to ejaculate and poor semen quality. Vibratory penile stimulation can induce ejaculation in many men with spinal cord injuries, but few studies have reported the effects of repeated vibratory stimulation on quantitative semen analysis tests. Fourteen spinal cord-injured males were enrolled in a prospective study to develop a quantitative profile of antegrade and retrograde ejaculate specimens collected during repeated vibratory stimulation. Antegrade specimens were obtained in 51 (84%) and retrograde specimens were obtained in all of the 61 trials (100%). Sperm were present in 70% and 64% of the antegrade and retrograde samples, respectively. Of the patients who underwent repeated vibratory stimulation attempts, three showed marked improvement in sperm counts and characteristics with repeated vibration and five showed normal fluctuation or no change in semen quality. Overall, there was a trend toward higher sperm counts in the antegrade samples (mean, 74.1 million) than in the retrograde samples (mean, 40 million), but no statistical significance was found. Mean sperm motility, sperm progressive motility, and percent normal sperm morphology were not significantly different between antegrade and retrograde specimens. Fructose and zinc, semen markers that reflect the presence of a true ejaculate, were present in all antegrade and retrograde specimens. In conclusion, (1) both antegrade and retrograde ejaculation occur with vibratory penile stimulation, with antegrade samples containing greater numbers of sperm but retrograde samples also having significant sperm counts, (2) repeated vibration may increase sperm concentration or may result in only minimal changes reflective of normal fluctuations, and (3) vibratory stimulation results in true ejaculation, as evidenced by semen markers.

Adult↗
PubMedDate not supplied

Addition of specific metabolites to bovine epididymal cell culture medium enhances survival and motility of cryopreserved sperm.

We have developed a cell culture system of bovine epididymal epithelium in which cryopreserved bovine sperm motility was efficiently maintained for many hours. The culture conditions to maintain viable epididymal cells are quite different from conditions normally used to incubate sperm cells. Thus, we have modified a previously described principal cell medium (PCM; Moore et al, 1992) using HEPES as a buffer and supplemented media with myo-inositol, pyruvate, lactate, glycerol, and carnitine to mimic epididymal intraluminal conditions. In the first experiments the effects of PCM and our epididymal cell medium (ECM) on sperm motility were compared in the absence of cells and evaluated by microscopic analysis under a phase contrast microscope or using the Hamilton Thorn Image Analyzer System. Our results showed that motility of cauda epididymal sperm was significantly higher in ECM than in PCM during a 48-hour incubation period when both media were supplemented with 10% fetal bovine serum (FBS). We then replaced FBS with bovine serum albumin (BSA) or no proteins at all to verify if ECM was able to enhance sperm survival. To test this aspect we used frozen-thawed sperm, which survived up to 48 hours when sperm cells were coincubated with epididymal cell monolayers. Hence, PCM, ECM, and different media containing each metabolite of ECM were supplemented with 0.5% BSA to assess motility of thawed sperm after an incubation period of 6 hours. A positive effect on sperm motility was observed in all fresh and unconditioned media containing 1 mM pyruvate. Motion parameters were more efficiently maintained in all conditioned media than in unconditioned media. Our results showed, however, that pyruvate was almost completely oxidized or consumed by epididymal cells during preincubation of culture media. We conclude that motility of frozen-thawed bovine spermatozoa can be improved using a culture medium or a medium conditioned by epididymal cell cultures without carnitine but containing mainly pyruvate, inositol, glycerol, and lactate.

Animals↗
PubMed1997

Formation of reactive oxygen species by spermatozoa from asthenospermic patients: response to treatment with pentoxifylline.

PURPOSE: We determined the incidence of reactive oxygen species formation by spermatozoa from asthenospermic patients, and the relationship between reactive oxygen species formation and sperm motion parameters. We also assessed the efficacy of in vitro and in vivo pentoxifylline treatment of asthenospermic patients whose spermatozoa generated high reactive oxygen species levels. MATERIALS AND METHODS: Reactive oxygen species formation by spermatozoa from asthenospermic patients and fertile volunteers was measured by chemoluminescence. Reactive oxygen species formation by the sperm preparations was investigated without stimulation (steady state), or after stimulation with N-formyl-methionyl-leucyl-phenylalanine (f-MLP) or phorbol-12-myristate-13-acetate. Spermatozoa from 15 asthenospermic patients whose spermatozoa produced high levels of reactive oxygen species at steady state were treated in vitro with pentoxifylline to determine its effect on reactive oxygen species generation and sperm motion parameters. These same 15 patients and 18 with asthenospermia whose spermatozoa did not produce reactive oxygen species at steady state were treated with pentoxifylline at 2 different dosages (300 and 1,200 mg. daily) to determine its effect on reactive oxygen species generation, sperm motion parameters and sperm fertilizing ability in vivo. RESULTS: When reactive oxygen species formation was detected in the steady state that was not stimulated by f-MLP, the source of reactive oxygen species could be attributed to the spermatozoa themselves. Spermatozoa from 15 of 71 asthenospermic patients generated reactive oxygen species at steady state. Pentoxifylline decreased reactive oxygen species generation by spermatozoa in these patients, and preserved the decrease of curvilinear velocity and beat cross frequency for 6 hours in vitro. For these patients orally administered pentoxifylline failed to decrease reactive oxygen species generation by spermatozoa, and had no effect on sperm motility, sperm motion parameters and sperm fertilizing ability at low dosage (300 mg. daily). However, it increased motility and beat cross frequency at high dosage (1,200 mg. daily) but it had no effect on sperm fertilizing ability. CONCLUSIONS: Stimulation of sperm preparations with f-MLP can identify the source of reactive oxygen species generated at steady state. Among asthenospermic patients there were some whose spermatozoa produced detectable steady state levels of reactive oxygen species. In this group pentoxifylline appeared to be effective for decreasing reactive oxygen species formation and preserving sperm motion parameters in vitro. Orally administered pentoxifylline had no effect at low dosage but it increased sperm motility and some sperm motion parameters without altering sperm fertilizing ability at high dosage.

Fertilization↗
PubMed1991

Extender and centrifugation effects on the motility patterns of slow-cooled stallion spermatozoa.

Slow-cooled stallion spermatozoa, with and without seminal plasma removed by centrifugation, were diluted in Kenney's extender (KE) containing nonfat dry skim milk with glucose and antibiotics or in KE supplemented by adding a modified high-potassium Tyrode's medium (KMT). Four ejaculates from each of four stallions were collected and divided factorially across these four treatments. Percentage of motile sperm, path velocity, and linearity immediately after treatment (0 h) and after storage at 4 degrees C for 24, 48, and 72 h were evaluated objectively by use of a HTM-2030 sperm motility analyzer. Stallions were a significant source of variation (P less than .01) throughout. After sperm had cooled, effects of stallion, extender, centrifugation, and their interactions were all found to be significant (P less than .01). The motility at 0, 24, 48, and 72 h for centrifuged KE was 74, 47, 39, and 24%; for uncentrifuged KE was 76, 56, 50, and 37%; for centrifuged KMT was 76, 75, 72, and 64%; and for uncentrifuged KMT was 80, 50, 26, and 13%, respectively. The extender x centrifugation interaction, after 24, 48, and 72 h of storage, accounted for half or more of the variation. Whereas centrifugation of semen extended in KE seemed to be harmful to sperm, motility of sperm extended in KMT after centrifugation was remarkably conserved for 72 h and was superior to all other treatments (P less than .05). This extender is promising for preserving liquid stallion semen when it must be transported before use in artificial insemination.

Analysis of Variance↗
PubMed1996

Characterization of in-vitro spermicidal activity of chelating agent against human sperm.

The effect of ethylenediaminetetraacetic acid (EDTA), a cationic chelating agent, on sperm motility and sperm penetration through cervical mucus was evaluated. EDTA exerted a relatively mild dose-dependent effect on sperm motility. Total loss of sperm motility (EC100) was achieved at a concentration of about 5.0 mg/mL of EDTA, which is much greater than the 200 micrograms/mL concentration of Nonoxynol-9 (N-9). EDTA salts, such as EDTA-Na2 and EDTA-Na4, showed weak spermicidal activity, while EDTA-K2 and EDTA-Ca had no spermicidal activity at concentrations up to 5.0 mg/mL. EDTA appeared to decrease the in vitro percentage of motile sperm as a function of exposure time. EDTA completely impeded the penetration of sperm into cervical mucus at a concentration much lower than that for 100% inhibition of sperm motility in semen. In the presence of 500-3200 micrograms/mL extra calcium, the first-order rate constant for sperm immobilization, which was derived from the slopes of a relative sperm motility curve, was reduced by 38.9--55.6% of the control and was inversely proportional to calcium concentration. After exposure to EDTA, the calcium ion concentration in semen was reduced as a function of EDTA concentration, which yielded a linear reduction of sperm motility. A decrease of approximately 65% in calcium ion concentration in semen seemed to be the threshold at which total loss of sperm motility occurred. Calcium ion was shown to play an essential role in regulating sperm molitility in semen, and EDTA was demonstrated to exert spermicidal activity by modulating calcium ion concentration in semen.

Animals↗
PubMed1988

Wheat germ agglutinin treatment of follicle cell-free mouse oocytes inhibits fertilization.

Controversy exists whether treatment of follicle cell-free oocytes with wheat germ agglutinin (WGA) prevents fertilization. Lack of inhibition in one case has led to the suggestion that acrosin may not be a zona lysin. To re-examine the effect of the WGA, the zona pellucida of follicle cell-free mouse oocytes was made more resistant to proteinase digestion by treatment with 10 or 50 micrograms/ml WGA. Such WGA-treated oocytes showed decreased fertilizability when washed to remove excess WGA and incubated with capacitated spermatozoa. Oocyte cleavage was used as an end point, because a large number of spermatozoa adhered to the eggs after WGA treatment, making observation of sperm penetration and pronucleus formation unreliable. Resistance to proteinase digestion increased, and the fertilizability decreased with the higher amount of WGA. The action of WGA was most likely not mediated by a direct effect on sperm motility, sperm acrosin activity, sperm binding to the zona pellucida, or oocyte cleavage. WGA did not affect the acrosome reaction of guinea pig spermatozoa. These data show that WGA treatment of follicle cell-free mouse oocytes results in decreased fertilizability, possibly by rendering the zona pellucida more resistant to sperm proteinase digestion.

Acrosin↗
PubMed2005

In vitro penetration of fresh and vitrified swine oocytes by homologous spermatozoa using different incubation systems.

The present study consisted of two experiments. In the first one, ejaculates from four boars were used to compare in vitro penetration (IVP) rates of fresh and vitrified swine oocytes by homologous spermatozoa in four treatments: fresh oocytes in conventional incubation (CO2 incubator) (FC), vitrified oocytes in conventional incubation (VC), fresh oocytes in submarine (bag) incubation (FS) and vitrified oocytes in submarine incubation (VS). The IVP rates for FC, VC, FS and VS were 46.5, 44.3, 36.9 and 33.1%, respectively. Analysis through Chi-square tests identified no differences in IVP rates between FC and VC and between FS and VS (P > 0.05), but IVP rate for FC was greater (P < 0.05) than those for both FS and VS. Besides IVP rate for VC did not differ (P > 0.05) from those for FC and FS, but it was greater than that for VS (P < 0.05). Logistic regression analysis identified differential effects of treatments dependant on individual boars. The second experiment evaluated the influence of semen storage period on the semen quality of the two boars associated with greater IVP rate in the first experiment. Semen quality was estimated by IVP rate using the VC treatment and by the following methods: sperm motility, sperm morphology, hypoosmotic swelling test (HOST) and thermal stress test (TST). According to analysis using Chi-square tests, IVP rate did not differ (P > 0.05), for the first boar, between 0 (100.0%) and 24 h of semen storage (98.1%) nor after 48 and 72 h (66.0 and 59.3%, respectively), but IVP rates were greater during the 0-24 h period compared with the 48-72 h period (P < 0.05). For the second boar, IVP rate at 0 h (50.6%) was greater (P < 0.05) than at 24, 48 and 72 h of semen storage (34.3, 28.3 and 24.0%, respectively), with no further differences observed after 24 h (P > 0.05). Logistic regression analysis identified that the effect of storage on IVP rate was influenced by the effect of individual boars. No differences in semen quality during the storage period were identified by conventional methods of semen evaluation, for either boar (P > 0.05) using analysis of variance with repeated measures. These results indicate that IVP test can be used to estimate boar fertility, even when vitrified oocytes are used (if using conventional CO2 incubators) or using an alternative submarine incubation system (if using fresh oocytes). The IVP test was the only method of semen evaluation that identified the reduction in semen quality up to 72 h of storage.

Analysis of Variance↗
PubMed2002

Penetration, adhesion, and fusion in mammalian sperm-egg interaction.

Fertilization is the sum of the cellular mechanisms that pass the genome from one generation to the next and initiate development of a new organism. A typical, ovulated mammalian egg is enclosed by two layers: an outer layer of approximately 5000 cumulus cells and an inner, thick extracellular matrix, the zona pellucida. To reach the egg plasma membrane, sperm must penetrate both layers in steps requiring sperm motility, sperm surface enzymes, and probably sperm-secreted enzymes. Sperm also bind transiently to the egg zona pellucida and the egg plasma membrane and then fuse. Signaling in the sperm is induced by sperm adhesion to the zona pellucida, and signaling in the egg by gamete fusion. The gamete molecules and molecular interactions with essential roles in these events are gradually being discovered.

Acrosome Reaction↗
PubMed1990

Tamoxifen and kallikrein in therapy of oligoasthenozoospermia: results of a randomized study.

Because of the good results achieved with tamoxifen in the treatment of oligozoospermia and with kallikrein in the treatment of asthenozoospermia, a randomized study of the combined treatment of oligoasthenozoospermia suggested itself. 67 patients with idiopathic normogonadotropic oligoasthenozoospermia were treated with 30 mg tamoxifen/day (n = 33) or with 30 mg tamoxifen/day and additionally 600 IU kallikrein/day (n = 34). It was shown that, apart from a significant increase in sperm density in both groups, the combination therapy also resulted in an overall significant increase (p less than 0.02) in sperm motility. Sperm morphology and the swell test remained unaffected. After 3 months of therapy, 4 pregnancies occurred in each group. In ejaculates with a sperm density of less than 10 million/ml not even one sperm parameter was significantly affected in any of the groups, while with an initial value of more than 10 million/ml the increase in motility was more significant (p less than 0.008) in the group with additional kallikrein therapy (n = 18). In the monotherapy group, no significant therapeutic effect on sperm motility was seen even in patients with a sperm density of more than 10 million. The combination of tamoxifen and kallikrein therefore seems to constitute an improvement of the systemic therapy of male subfertility in patients with moderately severe oligoasthenozoospermia.

Adult↗
PubMed1999

Effects of medium containing heparin and theophylline on capacitation and metabolic enzyme activities of ejaculated spermatozoa from dogs with asthenozoospermia.

The percentages of motile sperm (%MO), hyperactivated sperm (%HA), and acrosome-reacted sperm (%AR) of four beagle dogs with asthenozoospermia (AS) and five normal beagle dogs were determined during 7 h of incubation. The metabolic enzyme activities of the sperm was examined after 0 and 4 h of incubation. The sperm were incubated in canine capacitation medium (CCM) and CCM containing either 20 microg ml(-1) heparin (HE), 10 microg ml(-1) theophylline (TH) or 20 microg ml(-1) HE + 10 microg ml(-1) TH in glass tubes at 38 degrees C under 5% CO2 in air. The %HA and %AR were determined by counting the sperm exhibiting star-spin like movement and by the triple stain technique. The spermatozoa in HE + TH CCM were homogenized and centrifuged, and the metabolic activities of hexokinase, fructokinase, glucose-6-phosphodehydrogenase (G6PD), and pyruvate kinase in the sperm cytosol in the supernatant was measured with a spectrophotometer. The mean %MO and %HA values of both AS and normal dogs in the four types of CCM were highest in HE + TH CCM, with a mean %HA in HE + TH CCM of 78 +/- 5% (S.E.) after 7 h of incubation. However, there was little difference in %AR among the four types of CCM. The mean activities of the four enzymes in the sperm of AS dogs before incubation was significantly lower than in the sperm of normal dogs (P < 0.05, 0.01). However, after 4 h of incubation the activities of all enzymes in the sperm of both AS and normal dogs was clearly higher in HE + TH CCM than in the control CCM. These findings indicate that HE and TH in the medium are effective inactivating metabolic enzymes, maintaining longer sperm motility, and efficiently inducing HA even of the sperm of AS dogs.

Acrosome Reaction↗
PubMed1994

Low molecular weight components in bovine semen diffusate and their effects on motility of bull sperm.

Dialysis of diluted semen before cryopreservation is beneficial to sperm survival. This is due to removal of low molecular weight components from seminal plasma that are damaging to sperm. The apparent molecular weights (M(r)) of these components range between 1000 and 12,000 as estimated by gel permeation chromatography and electrophoresis. The detrimental effect on sperm motility is greatest with the components of M(r) between 5000 and 12,000 and with those of M(r) < 1500. Their effect on sperm motility was dependent on concentration. The small molecular weight components were derived from the high molecular weight components of seminal plasma through disaggregation under prescribed conditions.

Animals↗
PubMed1985

Reproductive criteria of beef bulls during and after exposure to increased ambient temperature.

Sixteen yearling Angus bulls were randomly assigned to one of two temperature-controlled chambers to determine the effects of elevated ambient temperature on body functions and semen characteristics. After 8 wk adjustment at 23 C, eight heat-stressed bulls were exposed to 35 +/- 1 C for 8 h and 31 +/- 1 C for 16 h during each 24-h period, and eight control bulls were maintained at 23 +/- 1 C for 8 wk. Then all bulls were exposed to 23 C for 8 wk. Bulls were fed so that both control and stressed bulls gained at similar rates (.58 kg/d). Semen was collected with an artificial vagina twice weekly before, during and after heat stress. During treatment, the respiratory rate of stressed bulls was greater (P less than .001) than that of control bulls (54.2 +/- 1.5, 29.9 +/- 1.5 breaths/min, respectively). Rectal temperatures were increased (P less than .01) from 38.2 +/- .1 to 38.7 +/- .1 C and water consumption was increased by 35% in stressed bulls when compared with controls. Semen volume was not altered by treatment, but percentage of motile sperm decreased (P less than .01) in stressed bulls by 2 wk after the start of heat treatment. Sperm motility of stressed bulls returned to normal values 8 wk after the end of heat treatment. Similarly, the percentage of aged acrosomes on sperm from stressed bulls increased (P less than .01) by the second week of treatment and remained greater than that of controls throughout the stress period.(ABSTRACT TRUNCATED AT 250 WORDS)

Acrosome↗