[Sperm motility and prostaglandins F2a, E1 AND E2].
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This study was carried out to monitor the effect of oral supplementation of vitamin C on various semen parameters in oligospermic, infertile, otherwise healthy individuals. Various semen parameters, including sperm motility, sperm count, and sperm morphology, were studied before and after the vitamin C treatment. A total of 13 infertile patients were included. Their ages ranged between 25 and 35 years. They had no genital infection or varicocele. Physical examination and other routine laboratory investigations were normal. General semen analysis revealed oligozoospermia (mean sperm count was 14.3 +/- 7.38 x 10(6) sperms/mL, mean sperm with normal morphology was 43 +/- 7.87%, and mean sperm motility was 31.2 +/- 9.61%). Testicular biopsy was not done. These patients received in an open trial of 1,000 mg of vitamin C twice daily for a maximum of 2 months. Results showed that the mean sperm count was increased to 32.8 +/- 10.3 x 10(6) sperms/mL (P < .001) after 2 months of vitamin C intake. The mean sperm motility was increased significantly to 60.1 +/- 8.47% (P < .001), and mean sperms with normal morphology increased significantly to 66.7 +/- 4.77% (P < .001). This study showed that vitamin C supplementation in infertile men might improve sperm count, sperm motility, and sperm morphology and might have a place as an additional supplement to improve the semen quality towards conception.
In a solution containing less than 5% seminal fluid, human sperm has a tendency to attach its head on the smooth surface of an object. Based on such a phenomenon, a simple method was developed to fix sperm head on the surface of glass slide. This method was demonstrated by studying caffeine effect on the tail movement of human sperm in phosphate buffered saline. By analysing the images recorded on a videocassette, caffeine, a well known stimulator of human sperm motility, was shown to induce an immediate increase of sperm TBF (tail beat frequency) and the amplitude of TBF increase was found to be larger in sperm with lower basal TBF value. This method can provide a flock of head-fixed motile sperm for a precise sperm tail movement assessment or for other purposes.
In recent studies, we found that the ectopic testis from postpuberal boars with unilateral abdominal cryptorchidism does not produce sperm. Therefore, in these males, the seminal characteristics can be used as indicators of the activity of the scrotal testis and its epididymis and also the accessory glands. The semen quality (ejaculate volume, cell-rich fraction volume, sperm concentration, sperm vitality, sperm motility, sperm morphology and cephalic stability of spermatozoa) was evaluated in healthy postpuberal boars and in postpuberal boars with unilateral abdominal cryptorchidism on the right side. In comparison with the healthy boars, the unilateral abdominal cryptorchid boars showed a significant decrease of the ejaculate volume, sperm concentration and sperm motility. The low sperm concentration indicated that unilateral abdominal cryptorchidism severely impairs the sperm production of the scrotal testis. The decrease of ejaculate volume was attributed to an abnormal activity of the accessory glands. The alterations in sperm motility develop as a result of dysfunctions in the epididymal epithelium and/or the accessory glands. The sperm vitality, sperm morphology and cephalic stability of spermatozoa maintained normal values; therefore, at testicular level, despite the low sperm production, the germ cell differentiation is not disturbed. At epididymal level, the morphological maturation of spermatozoa is not altered.
A total of 36 semen ejaculates, six from each of three Holstein-Friesian bulls and three Murrah buffalo bulls, were frozen in tris citric acid-fructose-egg-yolk-glycerol diluent after 1 hour of equilibration to study the effect of various cooling rates (15, 30, 60 and 120 minutes from 10 degrees to 5 degrees C vs a control sample cooled for 120 minutes from 28 degrees to 5 degrees C) and thawing temperatures (40 degrees C 60 seconds , 60 degrees C 15 seconds and 80 degrees C 5 seconds ) on prefreeze and post-thaw sperm motility. Sperm motility differed significantly (P < 0.01) between various cooling rates in both the Holstein-Friesian bull semen and the Murrah buffalo semen at prefreezing, immediately post-thawing, and after 1 hour of post-thaw incubation at 38 degrees C. Post-thaw sperm motility and survival at 38 degrees C were significantly (P<0.01) higher in Holstein-Friesian bulls at 60 degrees C and 80 degrees C than at 40 degrees C (39.79+/-2.46% and 38.15+/-2.18% Vs 35.16+/-2.19%, and 20.22+/-2.14% and 19.05+/-2.05% vs 14.83+/-1.64%, respectively). In Murrah buffalo bulls the recovery percentage and survival rate increased significantly (P<0.01) with the increase in temperature from 40 degrees C to 80 degrees C (41.72+/-2.45%, 47.45+/-2.09% and 51.61+/-2.06%; and 9.22+/-1.47%, 11.79+/-1.63% and 12.27+/-1.53%, respectively). Prefreeze motility did not differ between cattle and buffalo bulls (64.97+/-1.08% Vs 67.11+/-0.89%, respectively) but post-thaw motility was significantly (P<0.01) higher in the buffalo (46.93+/- 1.39% Vs 37.70+/-1.32%). While incubation survival was higher in the cattle (18.04+/-1.16% Vs 10.96+/-0.89%). A fast cooling rate was found to be detrimental for cattle spermatozoa, whereas the post-thaw buffalo sperm motility deteriorated very quickly at 38 degrees C. The influence of species-by-cooling rate interaction was significant (P<0.01) for post-thaw motility and survival rate, but the species-by-thawing or cooling-by-thawing interactions were not significant. These results suggest that a cooling rate of 2 hour either at 10 degrees C or 28 degrees C is essential for cattle semen. However, buffalo semen can be frozen successfully after 30 minutes of cooling at 10 degrees C. A thawing temperature of 60 degrees C yielded a higher sperm motility rate than 40 degrees C. Thus, our findings can be applied under tropical conditions for the successful freezing-thawing of bovine semen provided conception rates are not affected adversely.
Treatment of cancers with cytotoxic agents such as alkylating drugs often, but not always results in transient to permanent testicular dysfunction. The present study was planned to investigate the effects of dacarbazine [5-(3,3-dimethyltriazeno) imidazole-4-carboxamide] on testicular function in mice. Swiss albino mice (9-12 weeks old) were treated with 0, 5, 25, 50, or 100mg/kg body weight/day dacarbazine (i.p.) for 5 days at intervals of 24h between treatments. Mice were sacrificed on days 7, 14, 21, 28, 35, 49, and 70 after the last treatment (6 mice/dose/sample time), and the epididymal sperm count, sperm motility, sperm morphology, testicular histopathology (qualitative histopathology, seminiferous tubular diameter and epithelial height), and intra-testicular levels of testosterone and lactate dehydrogenase were assessed. Dacarbazine decreased the body weight only on day 28 at 25mg/kg dose-level, but increased the paired testes weights at 50mg/kg on day 7, at 25-100mg/kg on day 14, and at 25 and 50mg/kg on day 21 (P<0.05-0.01; one-way ANOVA and Bonferroni's post hoc test). The sperm count was decreased on all sampling days except at 5 and 25mg/kg dose-levels on day 70, but with severe oligospermia on days 28 and 35 (P<0.05-0.001). The sperm motility was decreased at 100mg/kg on days 14 and 21, at 5, 25, and 100mg/kg on day 28, and at all dose-levels on day 35 (P<0.05-0.001). Dacarbazine induced both head and tail abnormalities and some sperms with cytoplasmic droplets, but significant increase was seen in all dose groups on days 14 and 21, and at 100mg/kg dose-level on day 35. Drug-induced epithelial sloughing was seen on days 14-35 and other histopathological changes observed were vacuoles and abnormal cells. The STD was increased at 25-100mg/kg on day 7, at all dose-levels on day 14, at 50-100mg/kg on days 21 and 28, but without any effects on days 35-70 (P<0.05-0.001), and the tubular lumen was found dilated. The SE was increased on days 7, 21 and 28 at 100mg/kg and on day 14 at 50-100mg/kg. Dacarbazine reduced the intra-testicular testosterone level at 100mg/kg on day 7, at 5, 50 and 100mg/kg on day 14, at all dose-levels on days 21, 28, and 35, and at 50mg/kg on day 49 (P<0.05-0.001). The intra-testicular lactate dehydrogenase concentration increased at all dose-levels up to day 35, but without any effect on days 49 and 70 (P<0.05-0.001). There was no particular dose-response of dacarbazine on any parameters tested. The sperm count (except on day 7-positive correlation; Pearson product moment correlation) or sperm motility did not have any relation but increase in abnormal sperms showed negative correlation with decrease in testosterone level on days 7, 21 and 28. Decrease in sperm count was in negative correlation on days 14 and 35, and increase in abnormal sperms showed positive correlation on day 35 with increase in LDH level. Finally, the decrease in sperm motility had no correlation with increase in abnormal sperm shapes. We conclude that dacarbazine is genotoxic and cytotoxic to the mouse testis in a transient fashion, and these effects are exerted along with decrease in testosterone and increase in lactate dehydrogenase levels in the testis.
A microfluidic device was designed with two parallel laminar flow channels where non-motile spermatozoa and debris would flow along their initial streamlines and exit one outlet, whereas motile spermatozoa had an opportunity to swim into a parallel stream and exit a separate outlet. Motile sperm samples were prepared with density gradient separation (n = 5). Sperm motility was assessed the following day after exposing aliquots to polydimethylsiloxane (PDMS) used to construct the device. There was no difference in sperm motility when compared with unexposed aliquots (P > 0.05). Unprocessed semen samples (n = 10) were placed in wider channels and sperm motility and strict morphology were assessed from sorted outlets. Sperm motility increased from 44 +/- 4.5% to 98 +/- 0.4% (P < 0.05) and morphology increased from 10 +/- 1.05% to 22 +/- 3.3% (P < 0.05) following processing. Finally, density gradient prepared samples (n = 6) containing 5 x 10(6) motile spermatozoa/ml and 50 x 10(6) round immature germ cells/ml were sorted and assessed in a similar fashion. The ratio of motile spermatozoa to round immature germ cells in the wide inlet (1:10) was significantly improved in the thin outlet (33:1) (P < 0.05). This microfluidic device provides a novel method for isolating motile, morphologically normal spermatozoa from semen samples without centrifugation. This technology may prove useful in isolating motile spermatozoa from oligozoospermic samples, even with high amounts of non-motile gamete and/or non-gamete cell contamination. A movie sequence showing streaming and sorting of spermatozoa may be purchased for viewing on the internet at www.rbmonline.com/Article/847 (free to web subscribers).
OBJECTIVE: To determine the effects of treatment with dithiothreitol (DTT) on seminal viscosity, consistency, and sperm motility, motion characteristics, and morphology. DESIGN: Prospective, in vitro study. SETTING: University teaching hospital. PATIENTS: Semen donors and patients of the infertility program. Experiment 1, n = 34; experiment 2, n = 82. Sufficient seminal volume for use was the only selection criterion. INTERVENTIONS: Experiment 1: semen was combined with 5.9 mM DTT solution 1:1 (vol/vol), combined with distilled water 1:1 (vol/vol), or untreated. Experiment 2: semen was combined with 5.9 mM DTT solution 1:1 (vol/vol) or untreated. MAIN OUTCOME MEASURES: Seminal consistency and viscosity, percentage of motile sperm, sperm motion characteristics (straight line velocity, curvilinear velocity, mean linearity, and angle of lateral head displacement), sperm concentration, and sperm morphology. RESULTS: Treatment of semen with DTT reduced seminal consistency and viscosity. Treatment with DTT had no effect on sperm concentration or percentage motile sperm. Sperm velocity was reduced by DTT treatment, particularly in semen that had normal initial consistency. Morphology of sperm in semen exhibiting abnormal initial consistency was unaffected by DTT. CONCLUSION: Dithiothreitol effectively induced liquefaction of nonliquefied semen in vitro and had minimal effects on the sperm motility, motion characteristics, and morphology. Evaluation of DTT effects on the chromatin and on other functional traits of human sperm must be conducted before its use can be advocated.
This study was designed to determine if the value obtained after multiplying motile sperm concentration by seminal fructose concentration, named "true corrected fructose", correlates with sperm motility in asthenozoospermic men. Forty-two male partners in infertile couples were studied. Men were treated with 100 mg daily of clomiphene citrate for 5 days. Blood and semen samples were collected before treatment and 24 h after the end of treatment. Serum testosterone, seminal fructose and sperm motility were measured in each subject. Corrected fructose (log. sperm concentration multiplied by seminal fructose), and true corrected fructose (log. motile sperm concentration multiplied by seminal fructose) values were calculated. Prevalence of asthenozoospermia was 42.85% (18 of 42). Prevalence of hypofunction of the seminal vesicles was 9.5% using seminal fructose as a marker; 40.5% using seminal corrected fructose as a marker and 47.6% using true corrected fructose as a marker of seminal vesicle function. Regression analysis showed a better coefficient of determination between true corrected fructose and motile sperm concentration (R2=0.20, p < 0.001) than with corrected fructose (R(2)=0.05, p < 0.1) or fructose concentration (R(2)=0.006, p < 0.5). Asthenozoospermia was observed in 22.7% of subjects with normal function of the seminal vesicles, and in 65% of men with low values of true corrected fructose (z=6.02, p < 0.0001). Multivariate analysis showed that sperm motility grade 3 improved after treatment with clomiphene if true corrected fructose increased (p < 0.002). In those men whose seminal vesicle function improved after clomiphene treatment, a reduction in the prevalence of asthenozoospermia from 50 to 28.6% (z=3.10, p < 0.002) was observed whereas in those whose seminal vesicles did not respond to clomiphene the prevalence of asthenozoospermia was not reduced (z=1.05; p: NS). In conclusion, true corrected fructose measurement relates with sperm motility in cases of asthenozoospermia. Asthenozoospermia improves with clomiphene treatment if seminal vesicle function improves after treatment.
Electroejaculate traits and circulating follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone concentrations were analyzed in adult leopard cats (Felis bengalensis), a rare felid species indigenous to east Asia. The ability of leopard cat sperm to bind and penetrate zona-free hamster ova and zona-intact domestic cat oocytes in vitro was examined as a means of testing sperm function. The influence of culture media [Biggers, Whitten, Whittingham (BWW) vs. modified Krebs Ringer bicarbonate (mKRB)], seminal plasma removal, and swim-up separation on sperm motility, sperm morphology, and oocyte penetration also were assessed. Sperm treatments included dilution of raw semen (DR), ejaculate centrifugation, and either resuspension (NS) or swim-up processing (SU). The percentage of oocytes penetrated (penetration rate) and the number of penetrated sperm/oocyte (penetration index) were determined. Ejaculates from each male consisted of at least a 50% sperm motility rating, and hormone concentrations in individual males were unrelated to any ejaculate trait measured concurrently on the same day. The SU technique improved (P less than 0.05) percent sperm motility and the proportion of structurally normal sperm compared to DR and NS treatments. Leopard cat spermatozoa were capable of binding to and penetrating hamster ova and domestic cat oocytes; however, penetration was influenced by culture medium and seminal processing. In the hamster assay, a higher (P less than 0.05) penetration rate and penetration index were achieved when mKRB was used for gamete incubation instead of BWW. NS processing also increased (P less than 0.05) overall penetration compared to DR and SU. In the cat oocyte assay, zona penetration rate was similar (P greater than 0.05) in the DR, NS, and SU aliquots; however, the zona penetration index was increased (P less than 0.05) by the NS compared to the DR and SU treatments. This study 1) provides baseline ejaculate and endocrine norms for the leopard cat, 2) demonstrates that leopard cat sperm undergo nuclear decondensation in hamster ova and penetrate zona-intact domestic cat oocytes, 3) indicates that seminal plasma removal enhances leopard cat sperm fertilizing ability and ovum penetration, and 4) suggests that heterologous oocyte penetration is effective for assessing factors influencing fertilization and sperm function in this nondomestic felid.
This study was carried out to determine the milt quality in male Mesopotamian spiny eel (Mastacembelus mastacembelus; Bank & Solender in Russell, 1794) during spawning season. Review was performed using 58 M. mastecembulus males captured from Atatürk Dam Lake in Turkey. Milt samples were collected and evaluated daily during sampling period. In collected milt, volume of milt, sperm motility percentage, the duration of forward motility, sperm concentration, milt production and milt pH were evaluated. On June 10, 2005, 72% of the males were spermiating, which increased to 100% in the middle of June and dropped to 72% by late June. Sperm motility percentage showed a significant improvement after beginning of spermiation (P<0.001). Sperm concentration changed significantly (P<0.001) in milt collected during sampling period. Duration of sperm motility increased significantly (P<0.001) from beginning (8.22 min) through end (37.33 min) of spermiation. Milt production was maximal on June 15th, decreasing from a mean value of 3.12-0.46 x 10(9)/ml. The high pH in milt was observed during the week of spermiation (P<0.001). In conclusion, the results show that season has a significant influence on semen quality in male M. mastecembulus corresponding to spawning season in hot arid. Milt of good quality was especially collected in the middle of June. Spawning season in this fish species was found to be short.
An instrument called the Optibreed, BullMate sperm quality analyzer (SQA) contains a densitometer for determining sperm cell concentration and an optical sensor to evaluate light deflections caused by sperm movement. Analysis of light deflections enables the generation of a value called the sperm quality index (SQI). The SQI represents the quality of a semen sample defined by sperm motility, concentration, viability and morphology. The SQA was compared to conventional, microscopic techniques for determining percent motile sperm and sperm concentration in bull semen samples and evaluated for its ability to classify bulls as satisfactory or unsatisfactory potential breeders. Semen samples were collected from 105 mature beef bulls by electroejaculation (day 1) and from 51 of the same bulls by internal artificial vagina (IAV) on day 2. SQI values were arranged into 20 categories in increments of 50units from 0 to 1000units. Percent motile sperm and sperm concentration values from both methods were significantly positively correlated (P<0.000) with respective r values of 0.82 and 0.80. A calculation of kappa to evaluate the differences in percent motile sperm generated by each system yielded a value of 0.20 and 0.54 for unweighted and weighted determinations, respectively. The intraclass correlation coefficient used to evaluate the reliability of sperm cell concentration determinations was 0.62 (P<0.05). SQI values generated on days 1 and 2 ranged from 0-994 to 0-906, respectively. Bulls were categorized as satisfactory or unsatisfactory potential breeders in all categories. The most appropriate SQI for determining whether a bull was a satisfactory or unsatisfactory potential breeder was 500 with respective sensitivity, specificity, positive predictive values (PPV) and negative predictive values (NPV) of 81, 65, 90.79 and 44.83%. In this experiment the BullMate SQA was not a reliable substitute for conventional semen analysis and was not useful for determining bull breeding soundness.
Cypermethrin is considered as one of the endocrine disruptors. Isoflavones play an important role in various physiological processes in the body. It has both estrogenic and antioxidant effects. The objective of this study was to evaluate the protective role of isoflavones (2 mg/kg B.W) on semen quality and plasma testosterone levels of male New Zealand White rabbits given sublethal dose (24 mg/kg BW every other day for 12 weeks) of cypermethrin. Results showed that treatment with cypermethrin caused a significant decreases (P < 0.05) in ejaculate volume, sperm concentration, total sperm output, sperm motility (%), total motile sperm per ejaculate (TMS), packed sperm volume (PSV), semen initial fructose and plasma testosterone. In addition, live body weight (LBW), dry matter intake (DMI) and relative weights of testes and epididymis were decreased. On the other hand, treatment with cypermethrin increased (P < 0.05) the numbers of abnormal and dead sperms, and initial hydrogen ion concentration (pH). Results indicated that the presence of isoflavones together with cypermethrin was capable to minimize its harmful effects. Treatment with isoflavones alone had positive effects on some semen characteristics in spite of it is considered as estrogen-like compound. Since it causes significant increases in libido (by decreasing the reaction time), PSV, sperm motility and TMS, while abnormal and dead sperm were reduced compared to control animals. Meanwhile, isoflavones had no negative effect on ejaculate volume, total sperm output, sperm concentration, initial fructose concentration, pH and plasma testosterone levels. Results demonstrated the beneficial influences of isoflavones in reducing the negative effects of cypermethrin on reproductive characteristics of mature male rabbits. Interestingly, data showed that isoflavones alone caused an improvement in some semen quality and had no negative effects on male fertility, and did not have negative effects on male fertility.
Thirty-two New Zealand White growing rabbits (eight-week old) were used to determine the effect of feeding acacia-based diets on semen characteristics, plasma testosterone, free radicals, seminal plasma enzymes and lipids. Rabbits were randomly assigned to four equal groups. The first group (control) was fed a basal diet only, and the other three groups were fed other three diets, as follows: 80% of basal diet+20% of acacia leaves (low), 60% of basal diet+40% of acacia leaves (medium) and 40% of basal diet+60% of acacia leaves (high), respectively for 32 week. Semen samples were collected throughout the last 12 week of the experimental period. Rabbits fed on different levels of Acacia showed no significant changes in libido (reaction time), ejaculate volume, sperm concentration, packed sperm volume and initial hydrogen ion concentration compared to control. However, low and/or medium levels of Acacia caused significant (P<0.05) increase in total sperm output (TSO), sperm motility (%), total motile sperm per ejaculate (TMS), normal sperm, total functional sperm fraction (TFSF), semen initial fructose, live sperm and plasma testosterone. On the other hand, high level of Acacia did not show any significant change in TSO, sperm motility (%), TMS, initial fructose, TFSF or testosterone, while live and normal sperm decreased. All levels of Acacia caused a significant decrease in the concentrations of thiobarbituric acid-reactive substances (TBARS) and an increase in the activity of glutathione S-transferease. The activities of lactate dehydrogenase (LDH), aminotransferases and phosphatases were significantly increased in seminal plasma of animals fed low or medium levels of Acacia. Seminal plasma total lipid, triglyceride, low density lipoprotein and free fatty acids were significantly (P<0.05) decreased in low or medium levels of Acacia. On the other hand, total cholesterol, percentage cholesterol (out of total lipids) and high density lipoprotein were significantly (P<0.05) increased in seminal plasma of rabbits fed on low or medium levels of Acacia. High level of Acacia did not cause any changes in the previous parameters. It can be concluded that up to 40% Acacia leaves could be used successfully and safely in the diet of rabbits without adversely affecting their reproductive performance under subtropical conditions.
We are developing a data base that will allow us to select endpoints that would be useful in the detection of reproductive toxicity in a multigenerational test. In this effort, carbendazim (MBC), a known reproductive toxicant, was administered to male and female rats from weaning, through puberty, gestation, and lactation. A similar study was conducted with hamsters. In rats, MBC was administered at 0, 50, 100, 200, or 400 mg/kg/day. Hamsters were dosed at 0 or 400 mg/kg/day. In the parent (P0) generation, landmarks of puberty were measured. In females, estrous cyclicity, litter size, the number of implants, organ weights, and histology were assessed. Our assessment of the male rat included organ weights, testicular and epididymal sperm counts, a quantitative measure of sperm motility, sperm morphology, testicular histology, and endocrine measures. The growth, viability, and reproductive function of the offspring (F1) were observed during a 4-month period of continuous breeding. In the P0 of both species. MBC did not alter pubertal development, growth, or viability. The reproductive potential of the rats treated with MBC at 200 and 400 mg/kg/day was reduced due to effects on sperm production and fetal viability. In the male rat, MBC treatment markedly altered sperm morphology, testicular and epididymal weights, and sperm numbers and testicular histology. Fertility, sperm motility, and hormonal levels were altered, primarily in the males with very low sperm counts. The ability to conceive did not appear to involve a female factor. In P0 female rats, MBC administration caused postimplantation losses in the high-dosage groups and a few malformed rat pups were found in the litters from the 100 and 200 treatment groups. MBC was less toxic to the hamster than the rat. The only reproductive effects induced by MBC treatment were on sperm measures. Fertility of the P0 generation and fetal and neonatal (F1) viability were not decreased by MBC administration. In the male rat, testis weight, sperm numbers in the cauda epididymis and testis and sperm morphology were sensitive to the effects of MBC. In females, counting implantation scars at necropsy was useful, as this information allowed us to confirm pregnancy and identify postimplantation losses induced by MBC administration.